PRIMER REPORTE DE LA PRESENCIA DE Diaphorina citri (HEMIPTERA: LIVIIDAE EN MANABÍ

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José Bernardo Navarrete Cedeño

2016-12-01

Full Text Available Diaphorina citri (Hemiptera: Liviidae, conocido como el psílido asiático de los cítricos, es considerada una plaga clave de la citricultura mundial, debido a que es vector de la bacteria “Candidatus Liberibacter”, agente causal de la enfermedad “Huanglongbing”, que tiene efectos letales sobre vegetales dentro de la familia Rutaceae. En este documento se reporta la presencia de D. citri infestando Murraya spp., un arbusto de la familia Rutaceae, en la zona urbana del cantón Portoviejo. Este es el primer reporte de la plaga en la provincia de Manabí-Ecuador

Diaphorina citri nymphs are resistant to morphological changes induced by “Candidatus Liberibacter asiaticus” in midgut epithelial cells

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“Candidatus Liberibacter asiaticus” is the causative bacterium associated with citrus greening disease. “Ca. L. asiaticus” is transmitted by Diaphorina citri more efficiently when it is acquired by nymphs rather than adults. Why this occurs is not known. We compared midguts of D. citri reared on hea...

Disrupting Mating Behavior of Diaphorina citri (Liviidae).

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Lujo, S; Hartman, E; Norton, K; Pregmon, E A; Rohde, B B; Mankin, R W

2016-12-01

Severe economic damage from citrus greening disease, caused by 'Candidatus Liberibacter asiaticus' bacteria, has stimulated development of methods to reduce mating and reproduction in populations of its insect vector, Diaphorina citri (Hemiptera: Liviidae). Male D. citri find mating partners by walking on host plants, intermittently producing vibrational calls that stimulate duetting replies by receptive females. The replies provide orientational feedback, assisting the search process. To test a hypothesis that D. citri mating can be disrupted using vibrational signals that compete with and/or mask female replies, courtship bioassays were conducted in citrus trees with or without interference from female reply mimics produced by a vibrating buzzer. Statistically significant reductions occurred in the rates and proportions of mating when the buzzer produced reply mimics within 0.4 s after male courtship calls compared with undisturbed controls. Observations of courtship behaviors in the two bioassays revealed activity patterns that likely contributed to the reductions. In both disruption and control tests, males reciprocated frequently between structural bifurcations and other transition points where signal amplitudes changed. Males in the disruption bioassay had to select among vibrational signals combined from the buzzer and the female at each transition point. They often turned towards the buzzer instead of the female. There was a statistically significant reduction in the proportion of males mating if they contacted the buzzer, possibly due to its higher vibration amplitude and duration in comparison with female replies. Potential applications of D. citri mating disruption technology in citrus groves are discussed. Published by Oxford University Press on behalf of Entomological Society of America 2016. This work is written by US Government employees and is in the public domain in the US.

Behavioral responses of male Diaphorina citri (Hemiptera: Liviidae) to mating communication signals from vibration traps in citrus (Sapindales: Rutaceae) trees

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The Asian citrus psyllid, Diaphorina citri Kuwayama, vectors the bacterium causing citrus greening disease, which has devastated citrus production worldwide wherever it has been introduced. To help monitor and target D citri populations in commercial groves, thereby facilitating more effective manag...

Population diversity of Diaphorina citri (Hemiptera: Liviidae) in China based on whole mitochondrial genome sequences

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BACKGROUND: Diaphorina citri (Asian citrus psyllid, ACP) transmits “Candidatus Liberibacter asiaticus”, an unculturable alpha-proteobacterium associated with citrus Huanglongbing (HLB). ACP has been reported in 11 provinces/regions in China, yet its population diversity remains unclear. In this stud...

Spatiotemporal dynamics of the Southern California Asian citrus psyllid (Diaphorina citri) invasion.

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Bayles, Brett R; Thomas, Shyam M; Simmons, Gregory S; Grafton-Cardwell, Elizabeth E; Daugherty, Mathew P

2017-01-01

Biological invasions are governed by spatial processes that tend to be distributed in non-random ways across landscapes. Characterizing the spatial and temporal heterogeneities of the introduction, establishment, and spread of non-native insect species is a key aspect of effectively managing their geographic expansion. The Asian citrus psyllid (Diaphorina citri), a vector of the bacterium associated with huanglongbing (HLB), poses a serious threat to commercial and residential citrus trees. In 2008, D. citri first began expanding northward from Mexico into parts of Southern California. Using georeferenced D. citri occurrence data from 2008-2014, we sought to better understand the extent of the geographic expansion of this invasive vector species. Our objectives were to: 1) describe the spatial and temporal distribution of D. citri in Southern California, 2) identify the locations of statistically significant D. citri hotspots, and 3) quantify the dynamics of anisotropic spread. We found clear evidence that the spatial and temporal distribution of D. citri in Southern California is non-random. Further, we identified the existence of statistically significant hotspots of D. citri occurrence and described the anisotropic dispersion across the Southern California landscape. For example, the dominant hotspot surrounding Los Angeles showed rapid and strongly asymmetric spread to the south and east. Our study demonstrates the feasibility of quantitative invasive insect risk assessment with the application of a spatial epidemiology framework.

Spatiotemporal dynamics of the Southern California Asian citrus psyllid (Diaphorina citri invasion.

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Brett R Bayles

Full Text Available Biological invasions are governed by spatial processes that tend to be distributed in non-random ways across landscapes. Characterizing the spatial and temporal heterogeneities of the introduction, establishment, and spread of non-native insect species is a key aspect of effectively managing their geographic expansion. The Asian citrus psyllid (Diaphorina citri, a vector of the bacterium associated with huanglongbing (HLB, poses a serious threat to commercial and residential citrus trees. In 2008, D. citri first began expanding northward from Mexico into parts of Southern California. Using georeferenced D. citri occurrence data from 2008-2014, we sought to better understand the extent of the geographic expansion of this invasive vector species. Our objectives were to: 1 describe the spatial and temporal distribution of D. citri in Southern California, 2 identify the locations of statistically significant D. citri hotspots, and 3 quantify the dynamics of anisotropic spread. We found clear evidence that the spatial and temporal distribution of D. citri in Southern California is non-random. Further, we identified the existence of statistically significant hotspots of D. citri occurrence and described the anisotropic dispersion across the Southern California landscape. For example, the dominant hotspot surrounding Los Angeles showed rapid and strongly asymmetric spread to the south and east. Our study demonstrates the feasibility of quantitative invasive insect risk assessment with the application of a spatial epidemiology framework.

Diaphorina citri (Hemiptera: Liviidae) Abundance in Puerto Rico Declines with Elevation.

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Jenkins, David A; Hall, David G; Goenaga, Ricardo

2015-02-01

Diaphorina citri Kuwayama is the primary vector of Huanglongbing, the most devastating disease of citrus. D. citri populations in Puerto Rico were monitored with yellow sticky traps on citrus trees or other psyllid host plants at different elevations, ranging from 10 to 880 m above sea level. Trapping was conducted in March through May of 2013 and 2014 when psyllid populations usually are highest. Population levels of D. citri, based on the trapping data, varied among the sites, and there was a strong trend in both years for decreasing psyllid abundance with increased elevation based on the number of psyllids captured on traps and the proportion of trees shown to be infested. No psyllids were collected at an elevation of >600 m. Reduced populations at higher elevations could be a consequence of differences in temperature, air pressure, oxygen levels, ultraviolet light, or other factors alone or in combination. We discuss our results as they pertain to management of D. citri and Huanglongbing. Published by Oxford University Press on behalf of Entomological Society of America. 2015. This work is written by US Government employees and is in the public domain in the US.

Draft genome sequence of “Candidatus Liberibacter asiaticus” from Diaphorina citri in Guangdong, China

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The draft genome sequence of “Candidatus Liberibacter asiaticus” strain YCPsy from an Asian citrus psyllid (Diaphorina citri) in Guangdong of China is reported. The YCPsy strain has a genome size of 1,233,647 bp, 36.5% G+C content, 1,171 open reading frames (ORFs), and 53 RNAs....

Dispersion patterns and sampling plans for Diaphorina citri (Hemiptera: Psyllidae) in citrus.

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Sétamou, Mamoudou; Flores, Daniel; French, J Victor; Hall, David G

2008-08-01

The abundance and spatial dispersion of Diaphorina citri Kuwayama (Hemiptera: Psyllidae) were studied in 34 grapefruit (Citrus paradisi Macfad.) and six sweet orange [Citrus sinensis (L.) Osbeck] orchards from March to August 2006 when the pest is more abundant in southern Texas. Although flush shoot infestation levels did not vary with host plant species, densities of D. citri eggs, nymphs, and adults were significantly higher on sweet orange than on grapefruit. D. citri immatures also were found in significantly higher numbers in the southeastern quadrant of trees than other parts of the canopy. The spatial distribution of D. citri nymphs and adults was analyzed using Iowa's patchiness regression and Taylor's power law. Taylor's power law fitted the data better than Iowa's model. Based on both regression models, the field dispersion patterns of D. citri nymphs and adults were aggregated among flush shoots in individual trees as indicated by the regression slopes that were significantly >1. For the average density of each life stage obtained during our surveys, the minimum number of flush shoots per tree needed to estimate D. citri densities varied from eight for eggs to four flush shoots for adults. Projections indicated that a sampling plan consisting of 10 trees and eight flush shoots per tree would provide density estimates of the three developmental stages of D. citri acceptable enough for population studies and management decisions. A presence-absence sampling plan with a fixed precision level was developed and can be used to provide a quick estimation of D. citri populations in citrus orchards.

Host and environmental factors influencing "Candidatus Liberibacter asiaticus" acquisition in Diaphorina citri.

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Wu, Fengnian; Huang, Jiaquan; Xu, Meirong; Fox, Eduardo G P; Beattie, G Andrew C; Holford, Paul; Cen, Yijing; Deng, Xiaoling

2018-05-03

Diaphorina citri is a vector of "Candidatus Liberibacter asiaticus" (CLas) associated with citrus Huanglongbing. In this study, the infection and titers of CLas in the psyllid, were monitored for life cycle stage, sex, host-plant CLas titer, host-plant genotype, and ambient temperature. Acquisition efficiency of CLas by D. citri was highest in nymphs reared at 25 °C on a host plant with high CLas titers but was independent of the host genotypes assessed and of vector sex. We further observed that D. citri nymphs acquired CLas more rapidly than adults based on acquisition access periods (AAPs). CLas did not multiply in the alimentary canal, hemolymph, and salivary glands of adults for 18 d after a 3-day AAP as adult. However, CLas multiplication was detected in hemolymph and salivary gland of adults after the bacterium was acquired by nymphs. Eighty percent of salivary glands of adults contained CLas 18 d after a 3-day AAP as nymph compared to 10% 18 d after a 3-day AAP as adults. Different factors tested herein influenced CLas acquisition efficiency of D. citri, CLas multiplication and spread inside the psyllid. These observations serve to better understand mechanisms of CLas infection in D. citri. This article is protected by copyright. All rights reserved.

Effects of atmospheric pressure trends on calling, mate-seeking, and phototaxis of Diaphorina citri (Hemiptera: Liviidae)

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Insects and other animals sometimes modify behavior in response to changes in atmospheric pressure, an environmental cue that can provide warning of potentially injurious windy and rainy weather. To determine if Diaphorina citri (Hemiptera: Liviidae) calling, mate-seeking, and phototaxis behaviors w...

Comparative proteomic analysis of hemolymph from uninfected and Candidatus Liberibacter asiaticus-infected Diaphorina citri.

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Gill, T A; Chu, C; Pelz-Stelinski, K S

2017-02-01

Hemolymph was characterized from Diaphorina citri adults infected with the phytopathogen, Candidatus Liberibacter asiaticus (CLas), and compared with that from uninfected psyllids. This study identified 5531 and 3220 peptides within infected and uninfected hemolymph using nano-LC-MS/MS. A reduced number of proteins were detected for D. citri and all known endosymbionts within infected hemolymph as compared to uninfected hemolymph. A large number of immune defense proteins were absent from D. citri hemolymph; however, a single recognition protein (PGRP), two serine protease inhibitors, three prophenoloxidase (proPO) enzymes, and a single serine protease in an uninfected D. citri were detected. The hemolymph is nearly devoid of nutrient storage proteins. This is the first proteomic analysis of D. citri hemolymph that also analyses the components contributed by all the endosymbionts. By comparing the contribution of each endosymbiont (CCR, CPA, and WB) in the presence and absence of CLas infection, this study offers initial insights regarding the hemolymph response to microbial community shifts associated with D. citri infection status. Our data also present potential protein targets for analysis and disruption of CLas transmission that may facilitate management of huanglongbing (HLB) caused by CLas in citrus.

Whole-Genome Sequence of "Candidatus Profftella armatura" from Diaphorina citri in Guangdong, China.

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Wu, F; Deng, X; Liang, G; Huang, J; Cen, Y; Chen, J

2015-11-05

The genome of "Candidatus Profftella armatura" strain YCPA from Diaphorina citri in Guangdong, China, was sequenced. The strain has a chromosome of 457,565 bp, 24.3% G+C content, 364 predicted open reading frames (ORFs), and 38 RNAs, and a plasmid, pYCPA54, of 5,458 bp with 23.9% G+C content and 5 ORFs. Copyright © 2015 Wu et al.

Morphometric comparisons of Diaphorina citri (Hemiptera: Liviidae) populations from Iran, USA and Pakistan.

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Lashkari, Mohammadreza; Hentz, Matthew G; Boykin, Laura M

2015-01-01

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Liviidae), vector of citrus greening disease pathogen, Huanglongbing (HLB), is considered the most serious pest of citrus in the world. Prior molecular based studies have hypothesized a link between the D. citri in Iran and the USA (Florida). The purpose of this study was to collect morphometric data from D. citri populations from Iran (mtCOI haplotype-1), Florida (mtCOI haplotype-1), and Pakistan (mtCOI haplotype-6), to determine whether different mtCOI haplotypes have a relationship to a specific morphometric variation. 240 samples from 6 ACP populations (Iran-Jiroft, Chabahar; Florida-Ft. Pierce, Palm Beach Gardens, Port St. Lucie; and Pakistan-Punjab) were collected for comparison. Measurements of 20 morphological characters were selected, measured and analysed using ANOVA and MANOVA. The results indicate differences among the 6 ACP populations (Wilks' lambda = 0.0376, F = 7.29, P citri species in this study, (1) Iran, (2) USA (Florida) and (3) Pakistan population. Morphometric comparisons provided further resolution to the mtCOI haplotypes and distinguished the Florida and Iranian populations.

Effects of Temperature, Photoperiod, and Rainfall on Morphometric Variation of Diaphorina citri (Hemiptera: Liviidae).

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Paris, Thomson M; Allan, Sandra A; Hall, David G; Hentz, Matthew G; Croxton, Scott D; Ainpudi, Niharika; Stansly, Philip A

2017-02-01

Phenotypic plasticity provides a mechanism by which an organism can adapt to new or changing environments. Earlier studies have demonstrated the variability of Diaphorina citri Kuwayama (Asian citrus psyllid) population dynamics, but no analysis of morphological changes induced by seasonal or artificial laboratory-induced conditions has yet been documented. Such morphometric variation has been found to correspond in dispersal capabilities in several insect taxa. In this study, the effects of temperature and photoperiod on morphometric variation of D. citri were examined through laboratory rearing of psyllids under controlled temperatures (20 °C, 28 °C, and 30 °C) and under a short photoperiod of 10.5:13.5 (L:D) h and a long photoperiod of 16:8 (L:D) h. Diaphorina citri were field-collected monthly from three citrus groves in Fort Pierce, Gainesville, and Immokalee, FL, to evaluate potential field-associated environmental effects. Both traditional and geometric morphometric data were used to analyze the correlation between environmental and morphometric variation. A strong correlation was found between temperature and shape change, with larger and broader wings at colder temperatures in the laboratory. Short day length resulted in shorter and narrower wings as well. From the field, temperature, rainfall, and photoperiod were moderately associated with shape parameters. Adult D. citri with blue/green abdomens collected in the laboratory and field studies were larger in size and shape than those with brown/gray abdomens. Published by Oxford University Press on behalf of Entomological Society of America 2016. This work is written by US Government employees and is in the public domain in the US.

Killing Effects of an Isolated Serratia marcescens KH-001 on Diaphorina citri via Lowering the Endosymbiont Numbers.

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Hu, Wei; Kuang, Fan; Lu, Zhanjun; Zhang, Ning; Chen, Tingtao

2018-01-01

Huanglongbing (HLB) is the most devastating citrus disease worldwide, and suppression of the Asian citrus psyllid ( Diaphorina citri ) is regarded as an effective method to inhibit the spread of HLB. In this study, we isolated a strain named as Serratia marcescens KH-001 from D. citri nymphs suffering from disease, and evaluated its killing effect on D. citri via toxicity test and effect on microbial community in D. citri using high-throughput sequencing. Our results indicated that S. marcescens KH-001 could effectively kill 83% of D. citri nymphs, while the fermentation products of S. marcescens KH-001 only killed 40% of the D. citri nymphs. High-throughput sequencing results indicated that the S. marcescens KH-001 increased the OTU numbers from 62.5 (PBS buffer) to 81.5, while significantly lowered the Shannon index compared with Escherichia coli DH5α (group E) ( p citri endow S. marcescens KH-001 a sound killing effect on D. citri . Further work need to do before this strain is used as a sound biological control agents.

Diaphorina citri (Kuwayama, 1907 and Tamarixia radiata (Waterson, 1922 in citrus crops of Cundinamarca, Colombia

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Ebratt Ravelo Everth Emilio

2011-12-01

Full Text Available

The presence of Diaphorina citri Kuwayama (Hemiptera: Psyllidae and Tamarixia radiata (Waterston (Hymenoptera: Eulophidae was recorded in citrus crops of Cundinamarca, Colombia. This work is to know the geographic distribution of D. citri and an initial record of the parasitoid T. radiata in citrus producing areas of this department.

Resistance Management for Asian Citrus Psyllid, Diaphorina citri Kuwayama, in Florida.

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Chen, Xue Dong; Stelinski, Lukasz L

2017-09-20

The Asian citrus psyllid, Diaphorina citri Kuwayma, is one of the most important pests in citrus production. The objective of this study was to evaluate D. citri resistance management with three insecticide rotations and compare them with no rotation and an untreated check. The different insecticides (modes of action) tested were: dimethoate, imidacloprid, diflubenzuron, abamectin 3% + thiamethoxam 13.9%, and fenpropathrin. Eggs, nymph, and adult psyllids were counted weekly. Five insecticide applications were made in 2016. Insecticide susceptibility was determined by direct comparison with a laboratory susceptible population and field populations before and after all treatments were applied. Rankings of eggs, nymphs, and adults counted in treated plots were significantly lower than in the untreated control plots after each application. Initially, the resistance ratio (RR 50 ) for each rotation model, as compared with laboratory susceptible strain and the field population before application, was less than 5.76 and 4.31, respectively. However, after five applications with dimethoate, the RR 50 using the laboratory and pre-treatment field populations was 42.34 and 34.74, respectively. Our results indicate that effectively rotating modes of action can delay and/or prevent development of insecticide resistance in populations of D. citri .

Diaphorina citri (Hemiptera: Liviidae) Vector Competence for the Citrus Greening Pathogen 'Candidatus Liberibacter Asiaticus'.

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Tabachnick, Walter J

2015-06-01

Characterizing the vector competence of Diaphorina citri Kuwayama for 'Candidatus Liberibacter asiaticus,' the pathogen causing citrus greening, is essential for understanding the epidemiology of this disease that is threatening the U.S. citrus industry. Vector competence studies have been difficult because of the biology of D. citri, the inability to culture the pathogen, and the available diagnostic methods used to detect the bacteria in plant and insect tissues. The methods employed in many studies of D. citri vector competence may have overestimated amounts of live 'Ca. L. asiaticus' in both plant and insect tissues, and it is possible that the amounts of phloem ingested by psyllids may not contain sufficient detectable pathogen using current diagnostic methods. As a result of the difficulty in characterizing D. citri vector competence, the several daunting challenges for providing D. citri that are unable to inoculate 'Ca. L. asiaticus', as a novel method to control greening are discussed. Suggestions to overcome some of these challenges are provided. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Inter-Population Variability of Endosymbiont Densities in the Asian Citrus Psyllid (Diaphorina citri Kuwayama).

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Chu, Chia-Ching; Gill, Torrence A; Hoffmann, Mark; Pelz-Stelinski, Kirsten S

2016-05-01

The Asian citrus psyllid (Diaphorina citri Kuwayama) is an insect pest capable of transmitting Candidatus Liberibacter asiaticus (CLas), the causal agent of citrus greening in North America. D. citri also harbors three endosymbionts, Wolbachia, Candidatus Carsonella ruddii, and Candidatus Profftella armatura, which may influence D. citri physiology and fitness. Although genomic researches on these bacteria have been conducted, much remains unclear regarding their ecology and inter-population variability in D. citri. The present work examined the densities of each endosymbiont in adult D. citri sampled from different populations using quantitative PCR. Under field conditions, the densities of all three endosymbionts positively correlated with each other, and they are associated with D. citri gender and locality. In addition, the infection density of CLas also varied across populations. Although an analysis pooling D. citri from different populations showed that CLas-infected individuals tended to have lower endosymbiont densities compared to uninfected individuals, the difference was not significant when the population was included as a factor in the analysis, suggesting that other population-specific factors may have stronger effects on endosymbiont densities. To determine whether there is a genetic basis to the density differences, endosymbiont densities between aged CLas-negative females of two D. citri populations reared under standardized laboratory conditions were compared. Results suggested that inter-population variability in Wolbachia infection density is associated with the genotypes of the endosymbiont or the host. Findings from this work could facilitate understanding of D. citri-bacterial associations that may benefit the development of approaches for managing citrus greening, such as prevention of CLas transmission.

β-caryophyllene emitted from a transgenic Arabidopsis or chemical dispenser repels Diaphorina citri, vector of Candidatus Liberibacters

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Alquézar, B.; Volpe, H.X.L.; Magnani, R.F.; de Miranda, M.P.; Santos, M.A.; Wulff, N.A.; Bento, J.M.S.; Parra, J.R.P.; Bouwmeester, H.; Peña, L.

2017-01-01

Production of citrus, the main fruit tree crop worldwide, is severely threatened by Huanglongbing (HLB), for which as yet a cure is not available. Spread of this bacterial disease in America and Asia is intimately connected with dispersal and feeding of the insect vector Diaphorina citri,

Better Together: Association With 'Candidatus Liberibacter Asiaticus' Increases the Reproductive Fitness of Its Insect Vector, Diaphorina citri (Hemiptera: Liviidae).

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Pelz-Stelinski, K S; Killiny, N

2016-05-01

The duration of the evolutionary association between a pathogen and vector can be inferred based on the strength of their mutualistic interactions. A well-adapted pathogen is likely to confer some benefit or, at a minimum, exhibit low pathogenicity toward its host vector. Coevolution of the two toward a mutually beneficial association appears to have occurred between the citrus greening disease pathogen, Candidatus Liberibacter asiaticus (Las), and its insect vector, the Asian citrus psyllid, Diaphorina citri (Kuwayama). To better understand the dynamics facilitating transmission, we evaluated the effects of Las infection on the fitness of its vector. Diaphorina citri harboring Las were more fecund than their uninfected counterparts; however, their nymphal development rate and adult survival were comparatively reduced. The finite rate of population increase and net reproductive rate were both greater among Las-infected D. citri as compared with uninfected counterparts, indicating that overall population fitness of infected psyllids was improved given the greater number of offspring produced. Previous reports of transovarial transmission, in conjunction with increased fecundity and population growth rates of Las-positive D. citri found in the current investigation, suggest a long evolutionary relationship between pathogen and vector. The survival of Las-infected adult D. citri was lower compared with uninfected D. citri , which suggests that there may be a fitness trade-off in response to Las infection. A beneficial effect of a plant pathogen on vector fitness may indicate that the pathogen developed a relationship with the insect before secondarily moving to plants.

Dinamika Populasi Diaphorina citri Kuwayama (Homoptera: Psyllidae dan Deteksi CVPD dengan Teknik PCR

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I NYOMAN WIJAYA

2015-09-01

Full Text Available The research was conducted in the field of Taro village Gianyar regency and Katung village Bangli regency and at the Laboratory of Biotechnology Faculty of Agricultural Udayana University. The purpose of the research were to investigate population dynamic of Diaphorina citri Kuwayama and other insects that were assosiated with citrus cv. siam; and molecular detection of CVPD patogen in its vector, other phytophagous insects on citrus cv.siam and kemuning [Murraya paniculata (L. Jack.] The result of this study showed that in six month period there was one peak of D. citri population which was associated with the flushing period of citrus at Taro as well Katung villages. The peak of D. citri population of Taro village was higher than that of Katung village. Two species of parasitoids and four spesies of predators were found in the field and may contribute in the suppression of D. citri population. The parasitoids identified were Tamarixia radiata Wat. and Diaphorencyrtus alligharensi Shaffe, whereas the predators were Curinus coeruleus Mulsant, Phiddipus sp., Menochilus sexmaculatus Fabricius and Oxyopes javanus Thorell. T. radiata has been proven to be an effective nymph parasitoid that could suppress the population development of D. citri in citrus orchads. The research revealed that kemuning could serve as a source of infection because it is also a host of CVPD disease as well as of D. citri. Toxoptera citricidus Kirkaldy and Maleuterpes dentipes Hell. were not proven to contain CVPD disease.

First Record of Diaphorina citri (Hemiptera: Psyllidae) in Ecuador Infesting Urban Citrus and Orange Jasmine Trees

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Cornejo, J.F.; Chica, E.J.

2014-01-01

Abstract Adults and nymphs of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), were collected in the Guayaquil, Samborondón, and Durán cantons in coastal Ecuador. Psyllids were found in high numbers in citrus ( Citrus spp., Sapindales: Rutaceae) and orange jasmine ( Murraya exotica [L.] Jack, Sapindales: Rutaceae) trees within the Guayaquil-Samborondon-Duran conurbation; however, none was found during scoutings in the main citrus producing areas in coastal Ecuador. To the best of our knowledge, this is the first report of D. citri in Ecuador and the Pacific coastal plain of South America. PMID:25527601

Genetic diversity of Diaphorina citri and its endosymbionts across east and south-east Asia.

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Wang, Yanjing; Xu, Changbao; Tian, Mingyi; Deng, Xiaoling; Cen, Yijing; He, Yurong

2017-10-01

Diaphorina citri is the vector of 'Candidatus Liberibacter asiaticus', the most widespread pathogen associated huanglongbing, the most serious disease of citrus. To enhance our understanding of the distribution and origin of the psyllid, we investigated the genetic diversity and population structures of 24 populations in Asia and one from Florida based on the mtCOI gene. Simultaneously, genetic diversity and population structures of the primary endosymbiont (P-endosymbiont) 'Candidatus Carsonella ruddii' and secondary endosymbiont (S-endosymbiont) 'Candidatus Profftella armatura' of D. citri were determined with the housekeeping genes. AMOVA analysis indicated that populations of D. citri and its endosymbionts in east and south-east Asia were genetically distinct from populations in Pakistan and Florida. Furthermore, P-endosymbiont populations displayed a strong geographical structure across east and south-east Asia, while low genetic diversity indicated the absence of genetic structure among the populations of D. citri and its S-endosymbiont across these regions. The 'Ca. C. ruddii' is more diverse and structured than the D. citri and the 'Ca. P. armatura' across east and south-east Asia. Multiple introductions of the psyllid have occurred in China. Management application for controlling the pest is proposed based on the genetic information of D. citri and its endosymbionts. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Efficacy of essential oil of Piper aduncum against nymphs and adults of Diaphorina citri.

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Volpe, Haroldo Xl; Fazolin, Murilo; Garcia, Rafael B; Magnani, Rodrigo F; Barbosa, José Carlos; Miranda, Marcelo P

2016-06-01

Insecticide application is the main way to control Diaphorina citri. However, it causes environmental contamination, has a negative impact on beneficial organisms and leads to psyllid resistance. The essential oil of Piper aduncum has low toxicity towards the environment and contains dillapiol, which has proven to be effective against several crop pests. Here, we studied its efficacy against nymphs and adults of D. citri under laboratory conditions. Oils with three concentrations of dillapiol (69.3, 79.9 and 85.4%) at 0.5, 0.75 and 1.0% dilutions plus 0.025% adjuvant were tested. All treatments caused 90-100% mortality in nymphs. Topical treatments with oil containing 79.9 and 85.4% dillapiol at 0.75% and 1% dilutions were effective (mortality ≥80%) in adults. However, the essential oil showed no residual activity against adults (mortality ≤30%). Dillapiol-rich oil is a promising compound for D. citri control. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

Diverse Array of New Viral Sequences Identified in Worldwide Populations of the Asian Citrus Psyllid (Diaphorina citri) Using Viral Metagenomics.

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Nouri, Shahideh; Salem, Nidá; Nigg, Jared C; Falk, Bryce W

2015-12-16

The Asian citrus psyllid, Diaphorina citri, is the natural vector of the causal agent of Huanglongbing (HLB), or citrus greening disease. Together; HLB and D. citri represent a major threat to world citrus production. As there is no cure for HLB, insect vector management is considered one strategy to help control the disease, and D. citri viruses might be useful. In this study, we used a metagenomic approach to analyze viral sequences associated with the global population of D. citri. By sequencing small RNAs and the transcriptome coupled with bioinformatics analysis, we showed that the virus-like sequences of D. citri are diverse. We identified novel viral sequences belonging to the picornavirus superfamily, the Reoviridae, Parvoviridae, and Bunyaviridae families, and an unclassified positive-sense single-stranded RNA virus. Moreover, a Wolbachia prophage-related sequence was identified. This is the first comprehensive survey to assess the viral community from worldwide populations of an agricultural insect pest. Our results provide valuable information on new putative viruses, some of which may have the potential to be used as biocontrol agents. Insects have the most species of all animals, and are hosts to, and vectors of, a great variety of known and unknown viruses. Some of these most likely have the potential to be important fundamental and/or practical resources. In this study, we used high-throughput next-generation sequencing (NGS) technology and bioinformatics analysis to identify putative viruses associated with Diaphorina citri, the Asian citrus psyllid. D. citri is the vector of the bacterium causing Huanglongbing (HLB), currently the most serious threat to citrus worldwide. Here, we report several novel viral sequences associated with D. citri. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Resistance Management for Asian Citrus Psyllid, Diaphorina citri Kuwayama, in Florida

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Xue Dong Chen

2017-09-01

Full Text Available The Asian citrus psyllid, Diaphorina citri Kuwayma, is one of the most important pests in citrus production. The objective of this study was to evaluate D. citri resistance management with three insecticide rotations and compare them with no rotation and an untreated check. The different insecticides (modes of action tested were: dimethoate, imidacloprid, diflubenzuron, abamectin 3% + thiamethoxam 13.9%, and fenpropathrin. Eggs, nymph, and adult psyllids were counted weekly. Five insecticide applications were made in 2016. Insecticide susceptibility was determined by direct comparison with a laboratory susceptible population and field populations before and after all treatments were applied. Rankings of eggs, nymphs, and adults counted in treated plots were significantly lower than in the untreated control plots after each application. Initially, the resistance ratio (RR50 for each rotation model, as compared with laboratory susceptible strain and the field population before application, was less than 5.76 and 4.31, respectively. However, after five applications with dimethoate, the RR50 using the laboratory and pre-treatment field populations was 42.34 and 34.74, respectively. Our results indicate that effectively rotating modes of action can delay and/or prevent development of insecticide resistance in populations of D. citri.

Better Together: Association With ‘Candidatus Liberibacter Asiaticus’ Increases the Reproductive Fitness of Its Insect Vector, Diaphorina citri (Hemiptera: Liviidae)

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Pelz-Stelinski, K. S.; Killiny, N.

2016-01-01

The duration of the evolutionary association between a pathogen and vector can be inferred based on the strength of their mutualistic interactions. A well-adapted pathogen is likely to confer some benefit or, at a minimum, exhibit low pathogenicity toward its host vector. Coevolution of the two toward a mutually beneficial association appears to have occurred between the citrus greening disease pathogen, Candidatus Liberibacter asiaticus (Las), and its insect vector, the Asian citrus psyllid, Diaphorina citri (Kuwayama). To better understand the dynamics facilitating transmission, we evaluated the effects of Las infection on the fitness of its vector. Diaphorina citri harboring Las were more fecund than their uninfected counterparts; however, their nymphal development rate and adult survival were comparatively reduced. The finite rate of population increase and net reproductive rate were both greater among Las-infected D. citri as compared with uninfected counterparts, indicating that overall population fitness of infected psyllids was improved given the greater number of offspring produced. Previous reports of transovarial transmission, in conjunction with increased fecundity and population growth rates of Las-positive D. citri found in the current investigation, suggest a long evolutionary relationship between pathogen and vector. The survival of Las-infected adult D. citri was lower compared with uninfected D. citri, which suggests that there may be a fitness trade-off in response to Las infection. A beneficial effect of a plant pathogen on vector fitness may indicate that the pathogen developed a relationship with the insect before secondarily moving to plants. PMID:27418697

Pengaruh Jenis Mangsa dan Suhu pada Perkembangan Menochilus sexmaculatus Fabricius (Coleoptera: Coccinellidae dan Peranannya dalam Pengendalian Diaphorina citri Kuwayama (Hemiptera: Psyllidae

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Tris Haris Ramadhan

2008-07-01

Full Text Available Diaphorina citri Kuwayama (Hemiptera: Psyllidae is the vector of citrus greening (Huanglongbing bacterium and the most serious impediment to citrus culture. Classical biological control of this psyllid vector should contribute to suppress their population. This research was conducted to determine the performance of Menochilus sexmaculatus Fabricius (Coleoptera: Coccinellidae when they were fed with D. citri. The larval performance index of M. sexmaculatus on D. citri compared with Aphis craccivora Koch (Hemiptera: Aphididae diet was 1.3.M. sexmaculatus fed with D. citri had lower fitness than those fed with A. craccivora as shown by longer larval stadium, lower adult dry weight, less number of egg produced and lower percentage of egg hatched. M. sexmaculatus grew best at the temperature of 27oC. Employing the exclusion procedure under field condition,M. sexmaculatus could reduce the population of D. citri up to 90%. These findings showed that theM. sexmaculatus could be a potential predator in reducing D. citri, particularly when the more preferred prey A. craccivora was not present.   Diaphorina citri Kuwayama (Hemiptera: Psyllidae merupakan kelompok Psyllid yang menularkan penyebab penyakit Huanglongbing yang sangat berbahaya pada tanaman jeruk. Pengendalian hayati klasik telah banyak memberikan kontribusi dalam pengendalian di lapangan. Penelitian ini dilakukan untuk melihat penampilan Menochilus sexmaculatus Fabricius (Coleoptera: Coccinellidae dengan pakan D. citri. Indeks penampilan larva M. sexmaculatus dengan pakan D. citri dibandingkan dengan Aphis craccivora Koch (Hemiptera: Aphididae adalah 1,3. M. sexmaculatus yang diberi pakan D. citri menunjukkan penurunan kebugaran dibandingkan jika diberi pakan A. craccivora seperti yang ditunjukkan dengan stadium larva lebih lama, penurunan berat kering serangga dewasa, telur yang dihasilkan lebih sedikit, dan penurunan jumlah telur yang menetas. Menggunakan metode eksklusi pada kondisi

Effects on Asian Citrus Psyllid (Diaphorina citri Kuwayama) feeding behavior of fenpropathrin and chlorpyrifos within 24 hours of application

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Huanglongbing (HLB), also known as citrus greening disease, is one of the most destructive diseases affecting citrus production. The phloem-limited bacterium associated with HLB is transmitted by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae). One component of HLB managem...

Dynamic insecticide susceptibility changes in Florida populations of Diaphorina citri (Hemiptera: Psyllidae).

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Tiwari, Siddharth; Killiny, Nabil; Stelinski, Lukasz L

2013-02-01

Five field populations of Diaphorina citri Kuwayama from various regions of Florida were evaluated in 2011 for resistance to commonly used insecticides. Three diagnostic doses (LD50, LD75, and LD95), developed in 2009 using a laboratory susceptible population, were used to measure changes in susceptibility levels of field-collected populations as compared with a susceptible laboratory population. Further reductions in the susceptibility levels of D. citri to chlorpyriphos and fenpropathrin were determined, compared with results obtained in 2010. Mean percent mortality obtained from all five locations was significantly lower than observed with the laboratory susceptible population for all insecticides tested. Previously, expression of five CYP4 genes was implicated in contributing to insecticide metabolism in D. citri. In the current study, we compared the relative expression of these five CYP4 genes and their associated levels of protein expression among field-collected and laboratory susceptible populations. Expression of all CYP4 genes investigated was higher in field-collected populations when normalized against the laboratory susceptible population. There was an increased signal of a band corresponding to a 45 kDa protein in four of the five field populations as measured by the Western blot assay, which suggests increased production of cytochrome P450 enzymes. The current results indicate that insecticide resistance continues to increase in Florida populations ofD. citri, particularly to chlorpyriphos and fenpropathrin. However, there was no further decrease in susceptibility of Florida populations of D. citri to neonicotinoid insecticides in 2011 as compared with previous years.

Observations on the entomopathogenic fungus Hirsutella citriformis attacking adult Diaphorina citri (Hemiptera: Psyllid) in a managed citrus grove

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A two-year field study was conducted in an orange grove (0.7 ha) in Florida to characterize the phenology of the entomopathogen Hirsutella citriformis Speare infecting adults of the Asian citrus psyllid, Diaphorina citri Kuwayama. On the average over the two-year study, 23 percent of adults observed...

Biocuration and improvement of the Diaphorina citri draft genome assembly with long reads, optical maps and long-range scaffolding

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The Asian citrus psyllid (Diaphorina citri Kuwayama) is the insect vector of the bacterium Candidatus Liberibacter asiaticus (CLas), the causal agent for the citrus greening or Huanglongbing disease which threatens citrus industry worldwide. This vector is the primary target of approaches to stop th...

Chemical and behavioral analysis of the cuticular hydrocarbons from Asian citrus psyllid, Diaphorina citri.

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Mann, Rajinder S; Rouseff, Russell L; Smoot, Jack; Rao, Nandikeswara; Meyer, Wendy L; Lapointe, Stephen L; Robbins, Paul S; Cha, Dong; Linn, Charles E; Webster, Francis X; Tiwari, Siddharth; Stelinski, Lukasz L

2013-06-01

Huanglongbing (HLB) is the most destructive disease of citrus worldwide. The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the vector of the phloem-inhabiting bacterium, Candidatus Liberibacter asiaticus, which is presumed to cause HLB in Florida citrus. Laboratory and field studies were conducted to examine the behavioral responses of male and female D. citri to their cuticular extracts. In olfactometer assays, more male D. citri were attracted to one, five, or 10 female cuticular extract equivalent units than blank controls. The results were confirmed in field studies in which clear or yellow traps baited with 10 female cuticular extract equivalent units attracted proportionately more males than clear traps baited with male cuticular extract or unbaited traps. Analyses of cuticular constituents of male and female D. citri revealed differences between the sexes in chemical composition of their cuticular extracts. Laboratory bioassays with synthetic chemicals identified from cuticular extracts indicated that dodecanoic acid attracted more males than clean air. Traps baited with dodecanoic acid did not increase total catch of D. citri as compared with blank traps at the dosages tested; however, the sex ratio of psyllid catch was male biased on traps baited with the highest lure loading dosage tested (10.0 mg). © 2012 The Authors Insect Science © 2012 Institute of Zoology, Chinese Academy of Sciences.

Characterization of a Recombinant Cathepsin B-Like Cysteine Peptidase from Diaphorina citri Kuwayama (Hemiptera: Liviidae): A Putative Target for Control of Citrus Huanglongbing.

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Ferrara, Taíse Fernanda da Silva; Schneider, Vanessa Karine; Kishi, Luciano Takeshi; Carmona, Adriana Karaoglanovic; Alves, Marcio Fernando Madureira; Belasque-Júnior, Jose; Rosa, José César; Hunter, Wayne Brian; Henrique-Silva, Flávio; Soares-Costa, Andrea

2015-01-01

Huanglonbing (HLB) is one of the most destructive disease affecting citrus plants. The causal agent is associated with the phloem-limited bacterium Candidatus Liberibacter asiaticus (CLas) and the psyllid Diaphorina citri, vector of disease, that transmits the bacterium associated with HLB. The control of disease can be achieved by suppressing either the bacterium or the vector. Among the control strategies for HLB disease, one of the widely used consists in controlling the enzymes of the disease vector, Diaphorina citri. The insect Diaphorina citri belongs to the order Hemiptera, which frequently have cysteine peptidases in the gut. The importance of this class of enzymes led us to search for enzymes in the D. citri transcriptome for the establishment of alternatives strategies for HLB control. In this study, we reported the identification and characterization of a cathepsin B-like cysteine peptidase from D. citri (DCcathB). DCcathB was recombinantly expressed in Pichia pastoris, presenting a molecular mass of approximately 50 kDa. The enzyme hydrolyzed the fluorogenic substrate Z-F-R-AMC (Km = 23.5 μM) and the selective substrate for cathepsin B, Z-R-R-AMC (Km = 6.13 μM). The recombinant enzyme was inhibited by the cysteine protease inhibitors E64 (IC50 = 0.014 μM) and CaneCPI-4 (Ki = 0.05 nM) and by the selective cathepsin B inhibitor CA-074 (IC50 = 0.095 nM). RT-qPCR analysis revealed that the expression of the DCcathB in nymph and adult was approximately 9-fold greater than in egg. Moreover, the expression of this enzyme in the gut was 175-fold and 3333-fold higher than in the remaining tissues and in the head, respectively, suggesting that DCcathB can be a target for HLB control.

Characterization of a Recombinant Cathepsin B-Like Cysteine Peptidase from Diaphorina citri Kuwayama (Hemiptera: Liviidae: A Putative Target for Control of Citrus Huanglongbing.

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Taíse Fernanda da Silva Ferrara

Full Text Available Huanglonbing (HLB is one of the most destructive disease affecting citrus plants. The causal agent is associated with the phloem-limited bacterium Candidatus Liberibacter asiaticus (CLas and the psyllid Diaphorina citri, vector of disease, that transmits the bacterium associated with HLB. The control of disease can be achieved by suppressing either the bacterium or the vector. Among the control strategies for HLB disease, one of the widely used consists in controlling the enzymes of the disease vector, Diaphorina citri. The insect Diaphorina citri belongs to the order Hemiptera, which frequently have cysteine peptidases in the gut. The importance of this class of enzymes led us to search for enzymes in the D. citri transcriptome for the establishment of alternatives strategies for HLB control. In this study, we reported the identification and characterization of a cathepsin B-like cysteine peptidase from D. citri (DCcathB. DCcathB was recombinantly expressed in Pichia pastoris, presenting a molecular mass of approximately 50 kDa. The enzyme hydrolyzed the fluorogenic substrate Z-F-R-AMC (Km = 23.5 μM and the selective substrate for cathepsin B, Z-R-R-AMC (Km = 6.13 μM. The recombinant enzyme was inhibited by the cysteine protease inhibitors E64 (IC50 = 0.014 μM and CaneCPI-4 (Ki = 0.05 nM and by the selective cathepsin B inhibitor CA-074 (IC50 = 0.095 nM. RT-qPCR analysis revealed that the expression of the DCcathB in nymph and adult was approximately 9-fold greater than in egg. Moreover, the expression of this enzyme in the gut was 175-fold and 3333-fold higher than in the remaining tissues and in the head, respectively, suggesting that DCcathB can be a target for HLB control.

Population genetic structure of Diaphorina citri Kuwayama (Hemiptera: Liviidae): host-driven genetic differentiation in China.

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Meng, Lixue; Wang, Yongmo; Wei, Wen-Hua; Zhang, Hongyu

2018-01-24

The Asian citrus psyllid Diaphorina citri Kuwayama is a major pest in citrus production, transmitting Candidatus Liberibacter asiaticus. It has spread widely across eastern and southern China. Unfortunately, little is known about the genetic diversity and population structure of D. citri, making pest control difficult. In this study, nine specifically developed SSR markers and three known mitochondrial DNA were used for population genetics study of D. citri using 225 samples collected from all 7 distribution regions in China. Based on the SSR data, D. citri was found highly diverse with a mean observed heterozygosity of 0.50, and three subgroups were structured by host plant: (i) Shatangju, NF mandarin and Ponkan; (ii) Murraya paniculata and Lemon; (iii) Citrus unshiu, Bingtangcheng, Summer orange and Navel. No significant genetic differences were found with mtDNA data. We suggested the host-associated divergence is likely to have occurred very recently. A unimodal distribution of paired differences, the negative and significant Tajima's D and Fu's F S parameters among mtDNA suggested a recent demographic expansion. The extensive citrus cultivation and increased suitable living habitat was recommended as a key for this expansion event.

Cloning and expressing a highly functional and substrate specific farnesoic acid o-methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama)

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The Asian citrus psyllid, Diaphorina citri, transmits a phloem-limited bacterium, Candidatus Liberibacter asiaticus that causes citrus greening disease. Because juvenile hormone (JH) plays an important role in adult and nymphal development, we studied the final steps in juvenile hormone biosynthesis...

Survey of endosymbionts in the Diaphorina citri metagenome and assembly of a Wolbachia wDi draft genome.

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Surya Saha

Full Text Available Diaphorina citri (Hemiptera: Psyllidae, the Asian citrus psyllid, is the insect vector of Ca. Liberibacter asiaticus, the causal agent of citrus greening disease. Sequencing of the D. citri metagenome has been initiated to gain better understanding of the biology of this organism and the potential roles of its bacterial endosymbionts. To corroborate candidate endosymbionts previously identified by rDNA amplification, raw reads from the D. citri metagenome sequence were mapped to reference genome sequences. Results of the read mapping provided the most support for Wolbachia and an enteric bacterium most similar to Salmonella. Wolbachia-derived reads were extracted using the complete genome sequences for four Wolbachia strains. Reads were assembled into a draft genome sequence, and the annotation assessed for the presence of features potentially involved in host interaction. Genome alignment with the complete sequences reveals membership of Wolbachia wDi in supergroup B, further supported by phylogenetic analysis of FtsZ. FtsZ and Wsp phylogenies additionally indicate that the Wolbachia strain in the Florida D. citri isolate falls into a sub-clade of supergroup B, distinct from Wolbachia present in Chinese D. citri isolates, supporting the hypothesis that the D. citri introduced into Florida did not originate from China.

Survey of endosymbionts in the Diaphorina citri metagenome and assembly of a Wolbachia wDi draft genome.

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Saha, Surya; Hunter, Wayne B; Reese, Justin; Morgan, J Kent; Marutani-Hert, Mizuri; Huang, Hong; Lindeberg, Magdalen

2012-01-01

Diaphorina citri (Hemiptera: Psyllidae), the Asian citrus psyllid, is the insect vector of Ca. Liberibacter asiaticus, the causal agent of citrus greening disease. Sequencing of the D. citri metagenome has been initiated to gain better understanding of the biology of this organism and the potential roles of its bacterial endosymbionts. To corroborate candidate endosymbionts previously identified by rDNA amplification, raw reads from the D. citri metagenome sequence were mapped to reference genome sequences. Results of the read mapping provided the most support for Wolbachia and an enteric bacterium most similar to Salmonella. Wolbachia-derived reads were extracted using the complete genome sequences for four Wolbachia strains. Reads were assembled into a draft genome sequence, and the annotation assessed for the presence of features potentially involved in host interaction. Genome alignment with the complete sequences reveals membership of Wolbachia wDi in supergroup B, further supported by phylogenetic analysis of FtsZ. FtsZ and Wsp phylogenies additionally indicate that the Wolbachia strain in the Florida D. citri isolate falls into a sub-clade of supergroup B, distinct from Wolbachia present in Chinese D. citri isolates, supporting the hypothesis that the D. citri introduced into Florida did not originate from China.

The effects of host, geographic origin, and gender on the thermal requirements of Diaphorina citri (Hemiptera: Psyllidae).

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Nava, Dori E; Gomez-Torres, Mariuxi L; Rodrigues, Marjorie D; Bento, José M S; Haddad, Marinéia L; Parra, José R P

2010-04-01

Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is the vector of the bacteria that causes citrus greening and is considered one of the world's most important citrus diseases. We examined how host, geographic region, and gender affect the thermal requirements of D. citri. The insects were reared in climatic chambers at constant temperatures of 18, 20, 22, 25, 28, 30, and 32 +/- 1 degrees C, 70 +/- 10% RH, and a 14 h photophase. Host plants for D. citri included orange (Citrus sinensis [Rutaceae]) varieties Pêra and Natal, the rootstock, Rungpur lime (C. limonia [Rutaceae]) and the natural host, Orange jessamine (Murraya paniculata [Rutaceae]). To study the influence of geographic origin on thermal requirements, we studied D. citri populations from Piracicaba, SP (warmer region) and Itapetininga, SP (cooler region). The duration and survival of the development stages and the duration of the total development (egg-adult) did not differ significantly on the different hosts, but it did vary with temperature. Nymphs of D. citri created on the different hosts have the same thermal requirements. The thermal requirements for this species collected from the two climate regions were identical; males and females also had the same thermal requirements.

Induced resistance against the Asian citrus psyllid, Diaphorina citri, by β-aminobutyric acid in citrus.

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Tiwari, Siddharth; Meyer, Wendy L; Stelinski, Lukasz L

2013-10-01

β-Aminobutyric acid (BABA) is known to induce resistance to microbial pathogens, nematodes and insects in several host plant/pest systems. The present study was undertaken to determine whether a similar effect of BABA occurred against the Asian citrus psyllid, Diaphorina citri Kuwayama, in citrus. A 25 mM drench application of BABA significantly reduced the number of eggs/plant as compared with a water control, whereas 200 and 100 mM applications of BABA reduced the numbers of nymphs/plant and adults/plants, respectively. A 5 mM foliar application of BABA significantly reduced the number of adults but not eggs or nymphs when compared with a water control treatment. In addition, leaf-dip bioassays using various concentrations (25–500 mM) of BABA indicated no direct toxic effect on 2nd and 5th instar nymphs or adult D. citri. BABA-treated plants were characterized by significantly lower levels of iron, magnesium, phosphorus, sodium, sulfur and zinc as compared with control plants. The expression level of the PR-2 gene (β-1,3-glucanase) in BABA-treated plants that were also damaged by D. citri adult feeding was significantly higher than in plants exposed to BABA, D. citri feeding alone or control plants. Our results indicate the potential for using BABA as a systemic acquired resistance management tool for D. citri.

An assessment of interspecific competition between two introduced parasitoids of Diaphorina citri (Hemiptera: Liviidae) on caged citrus plants.

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Vankosky, Meghan A; Hoddle, Mark S

2017-06-07

Two parasitoids attacking nymphs of Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), Tamarixia radiata (Waterston) (Hymenoptera: Eulophidae) and Diaphorencyrtus aligarhensis (Shafee, Alam & Agarwal) (Hymenoptera: Encyrtidae) are being released in California, USA in a classical biological control program. To evaluate the effect of multiple parasitoid species on D. citri mortality, we conducted mesocosm experiments under controlled conditions using a complete block design with 6 treatments (D. citri nymphs exposed to: no parasitoids; D. aligarhensis or T. radiata alone; D. aligarhensis or T. radiata released first (by 48 h); and both species released simultaneously). Parasitism of D. citri nymphs by T. radiata exceeded 60% and was unchanged when D. aligarhensis were present. Parasitism by D. aligarhensis was greatest when T. radiata was absent (∼28%) and was reduced in all treatments with T. radiata present (citri mortality and parasitoid-related mortality of D. citri was consistent across parasitoid treatments. Laboratory results suggest that competition between D. aligarhensis and T. radiata is asymmetric and favors T. radiata. It may be difficult for D. aligarhensis to contribute significantly to D. citri biological control where T. radiata is present. However, results reported here suggest that competition between T. radiata and D. aligarhensis is not likely to reduce parasitism by T. radiata or reduce parasitoid-induced mortality of D. citri. © 2017 Institute of Zoology, Chinese Academy of Sciences.

PENDIDIKAN DAN PELATIHAN PENGENDALIAN KUTU LONCAT JERUK (Diaphorina citri Kuwayama SEBAGAI HAMA DAN VEKTOR PENYAKIT CVPD DI DESA TARO, GIANYAR

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Wijaya I N.

2012-10-01

Full Text Available Training on jumping plantlice of citrus (Diaphorina citri Kuw the vector of CVPD (Citrus Vein Phloem Degeneration has been held at Taro village, Gianyar regency on 30st July 2012. The activities aimed to know bioecology of D. citri, the vector of CVPD. The training was attended by 32 participants from local farmer groups of Taro village. The method used in activities were lectures, demontration and practical in the citrus field. All participants participited anthusiastically and hope the have the next intensive traning of the citrus culture.

Morphometric comparisons of Diaphorina citri (Hemiptera: Liviidae populations from Iran, USA and Pakistan

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Mohammadreza Lashkari

2015-05-01

Full Text Available The Asian citrus psyllid (ACP, Diaphorina citri Kuwayama (Hemiptera: Liviidae, vector of citrus greening disease pathogen, Huanglongbing (HLB, is considered the most serious pest of citrus in the world. Prior molecular based studies have hypothesized a link between the D. citri in Iran and the USA (Florida. The purpose of this study was to collect morphometric data from D. citri populations from Iran (mtCOI haplotype-1, Florida (mtCOI haplotype-1, and Pakistan (mtCOI haplotype-6, to determine whether different mtCOI haplotypes have a relationship to a specific morphometric variation. 240 samples from 6 ACP populations (Iran—Jiroft, Chabahar; Florida—Ft. Pierce, Palm Beach Gardens, Port St. Lucie; and Pakistan—Punjab were collected for comparison. Measurements of 20 morphological characters were selected, measured and analysed using ANOVA and MANOVA. The results indicate differences among the 6 ACP populations (Wilks’ lambda = 0.0376, F = 7.29, P < 0.0001. The body length (BL, circumanal ring length (CL, antenna length (AL, forewing length (WL and Rs vein length of forewing (RL were the most important characters separating the populations. The cluster analysis showed that the Iran and Florida populations are distinct from each other but separate from the Pakistan population. Thus, three subgroups can be morphologically discriminated within D. citri species in this study, (1 Iran, (2 USA (Florida and (3 Pakistan population. Morphometric comparisons provided further resolution to the mtCOI haplotypes and distinguished the Florida and Iranian populations.

Characterization of the voltage-gated sodium channel of the Asian citrus psyllid, Diaphorina citri.

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Liu, Bin; Coy, Monique R; Wang, Jin-Jun; Stelinski, Lukasz L

2017-02-01

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is an important insect pest of citrus. It is the vector of 'Candidatus' Liberibacter asiaticus, a phloem-limited bacterium that infects citrus, resulting in the disease Huanglongbing (HLB). Disease management relies heavily on suppression of D. citri populations with insecticides, including pyrethroids. In recent annual surveys to monitor insecticide resistance, reduced susceptibility to fenpropathrin was identified in several field populations of D. citri. The primary target of pyrethroids is the voltage-gated sodium channel (VGSC). The VGSC is prone to target-site insensitivity because of mutations that either reduce pyrethroid binding and/or alter gating kinetics. These mutations, known as knockdown resistance or kdr, have been reported in a wide diversity of arthropod species. Alternative splicing, in combination with kdr mutations, has been also associated with reduced pyrethroid efficacy. Here we report the molecular characterization of the VGSC in D. citri along with a survey of alternative splicing across developmental stages of this species. Previous studies demonstrated that D. citri has an exquisite enzymatic arsenal to detoxify insecticides resulting in reduced efficacy. The results from the current investigation demonstrate that target-site insensitivity is also a potential basis for insecticide resistance to pyrethroids in D. citri. The VGSC sequence and its molecular characterization should facilitate early elucidation of the underlying cause of an established case of resistance to pyrethroids. This is the first characterization of a VGSC from a hemipteran to this level of detail, with the majority of the previous studies on dipterans and lepidopterans. © 2015 Institute of Zoology, Chinese Academy of Sciences.

The genetic structure of an invasive pest, the Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae).

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Guidolin, Aline S; Fresia, Pablo; Cônsoli, Fernando L

2014-01-01

The Asian citrus psyllid Diaphorina citri is currently the major threat to the citrus industry as it is the vector of Candidatus Liberibacter, the causal agent of huanglongbing disease (HLB). D. citri is native to Asia and now colonizes the Americas. Although it has been known in some countries for a long time, invasion routes remain undetermined. There are no efficient control methods for the HLB despite the intensive management tools currently in use. We investigated the genetic variability and structure of populations of D. citri to aid in the decision making processes toward sustainable management of this species/disease. We employed different methods to quantify and compare the genetic diversity and structure of D. citri populations among 36 localities in Brazil, using an almost complete sequence of the cytochrome oxidase I (COI) gene. Our analyses led to the identification of two geographically and genetically structured groups. The indices of molecular diversity pointed to a recent population expansion, and we discuss the role of multiple invasion events in this scenario. We also argue that such genetic diversity and population structure may have implications for the best management strategies to be adopted for controlling this psyllid and/or the disease it vectors in Brazil.

The genetic structure of an invasive pest, the Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae.

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Aline S Guidolin

Full Text Available The Asian citrus psyllid Diaphorina citri is currently the major threat to the citrus industry as it is the vector of Candidatus Liberibacter, the causal agent of huanglongbing disease (HLB. D. citri is native to Asia and now colonizes the Americas. Although it has been known in some countries for a long time, invasion routes remain undetermined. There are no efficient control methods for the HLB despite the intensive management tools currently in use. We investigated the genetic variability and structure of populations of D. citri to aid in the decision making processes toward sustainable management of this species/disease. We employed different methods to quantify and compare the genetic diversity and structure of D. citri populations among 36 localities in Brazil, using an almost complete sequence of the cytochrome oxidase I (COI gene. Our analyses led to the identification of two geographically and genetically structured groups. The indices of molecular diversity pointed to a recent population expansion, and we discuss the role of multiple invasion events in this scenario. We also argue that such genetic diversity and population structure may have implications for the best management strategies to be adopted for controlling this psyllid and/or the disease it vectors in Brazil.

Behavioral plasticity in feeding by Diaphorina citri (Hemiptera, Liviidae): ingestion from phloem versus xylem is influenced by leaf age and surface

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Diaphorina citri is a major pest of citrus because it transmits the bacterium that causes Huanglongbing (HLB) (a.k.a. citrus greening). One approach to disease management is vector management using insecticides. However, knowledge of vector mortality alone is not sufficient if the vector has had tim...

Characterization of a recombinant Cathepsin B-Like cysteine peptidase from Diaphorina citri Kuwayama (Hemiptera: Liviidae): A putative target control of citrus huanglongbing

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Huanglongbing (HLB) is one of the most destructive disease affecting citrus plants. The causal agent is associated with the phloem-limited bacterium Candidatus Liberibacter asiaticus (CLas) spread by the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae). Among the control strategies for H...

Killing Effects of an Isolated Serratia marcescens KH-001 on Diaphorina citri via Lowering the Endosymbiont Numbers

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Wei Hu

2018-05-01

Full Text Available Huanglongbing (HLB is the most devastating citrus disease worldwide, and suppression of the Asian citrus psyllid (Diaphorina citri is regarded as an effective method to inhibit the spread of HLB. In this study, we isolated a strain named as Serratia marcescens KH-001 from D. citri nymphs suffering from disease, and evaluated its killing effect on D. citri via toxicity test and effect on microbial community in D. citri using high-throughput sequencing. Our results indicated that S. marcescens KH-001 could effectively kill 83% of D. citri nymphs, while the fermentation products of S. marcescens KH-001 only killed 40% of the D. citrinymphs. High-throughput sequencing results indicated that the S. marcescens KH-001 increased the OTU numbers from 62.5 (PBS buffer to 81.5, while significantly lowered the Shannon index compared with Escherichia coli DH5α (group E (p < 0.05. OTU analysis showed that the S. marcescens KH-001 had significantly reduced the relative abundance of endosymbionts Wolbachia, Profftella, and Carsonella in group S compared with that in other groups (p < 0.05. Therefore, the direct killing effect of the fermentation products of S. marcescens KH-001 and the indirect effect via reducing the numbers of endosymbionts (Wolbachia, Profftella, and Carsonella of D. citri endow S. marcescens KH-001 a sound killing effect on D. citri. Further work need to do before this strain is used as a sound biological control agents.

Living on the Edges: Spatial Niche Occupation of Asian Citrus Psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), in Citrus Groves.

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Sétamou, Mamoudou; Bartels, David W

2015-01-01

The spatial niche occupation of the Asian citrus psyllid, Diaphorina citri Kuwayama, 1908, was evaluated to determine its field colonization and food resource exploitation strategies in citrus groves. Mature grapefruit and sweet orange groves were surveyed as part of an area-wide program in 2009-2010 to determine D. citri population densities and between-tree distribution. In both cultivars, significantly more psyllids were found on perimeter trees throughout the study period suggesting a strong edge effect in D. citri distribution in the groves. D. citri densities and infestation levels gradually declined from the edge to the center of grove. Higher numbers of D. citri were recorded on trees located on the east and south sides of the groves than those on the west and north sides. Citrus groves located at the outer edge of the study with at least one side non-surrounded to other citrus groves harbored significantly more D. citri than groves located within the block cluster and entirely surrounded by other groves. In detailed field studies during 2012, infestation of D. citri started from border trees in the grove where possibly one generation is completed before inner trees become infested. In addition, psyllid densities decreased significantly with increasing distance from the grove edge. Using the selection index, D citri exhibited a strong niche occupation preference for border trees.

Living on the Edges: Spatial Niche Occupation of Asian Citrus Psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae, in Citrus Groves.

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Mamoudou Sétamou

Full Text Available The spatial niche occupation of the Asian citrus psyllid, Diaphorina citri Kuwayama, 1908, was evaluated to determine its field colonization and food resource exploitation strategies in citrus groves. Mature grapefruit and sweet orange groves were surveyed as part of an area-wide program in 2009-2010 to determine D. citri population densities and between-tree distribution. In both cultivars, significantly more psyllids were found on perimeter trees throughout the study period suggesting a strong edge effect in D. citri distribution in the groves. D. citri densities and infestation levels gradually declined from the edge to the center of grove. Higher numbers of D. citri were recorded on trees located on the east and south sides of the groves than those on the west and north sides. Citrus groves located at the outer edge of the study with at least one side non-surrounded to other citrus groves harbored significantly more D. citri than groves located within the block cluster and entirely surrounded by other groves. In detailed field studies during 2012, infestation of D. citri started from border trees in the grove where possibly one generation is completed before inner trees become infested. In addition, psyllid densities decreased significantly with increasing distance from the grove edge. Using the selection index, D citri exhibited a strong niche occupation preference for border trees.

Effects of buprofezin and diflubenzuron on various developmental stages of Asian citrus psyllid, Diaphorina citri.

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Tiwari, Siddharth; Clayson, Paul J; Kuhns, Emily H; Stelinski, Lukasz L

2012-10-01

Diaphorina citri populations in Florida are developing resistance to commonly used neurotoxic insecticides. Alternatives to neurotoxins, such as insect growth regulators, are needed to control this season-long subtropical pest to prevent or delay development of insecticide resistance. In the present investigation, two insect growth regulators (IGRs), buprofezin and diflubenzuron, were evaluated against various developmental stages of D. citri. The 0-1-day-old D. citri eggs were more susceptible to buprofezin and diflubenzuron than the 3-4-day-old eggs. Adult emergence was completely suppressed by treating first- or third-instar nymphs with buprofezin or diflubenzuron at 30-240 or 23-184 µg mL(-1) rates respectively. Treatment of fifth-instar nymphs with diflubenzuron at a rate of 184 µg mL(-1) and with buprofezin at 30-240 µg mL(-1) rates resulted in approximately 20 and 15-80% reductions in adult emergence respectively. The mean number of eggs per plant was reduced at 5 days after topical treatment with diflubenzuron. Mean egg hatch per plant was reduced at 5 and 6-15 days after topical treatments with buprofezin and diflubenzuron respectively. Buprofezin and diflubenzuron effectively suppressed D. citri adult emergence. D. citri were more susceptible as early (first-third-instar) than late (fifth-instar) nymphs. Both IGRs inhibited egg production and egg hatch. Reduction in the number of subsequent offspring suggests reduced vertical transmission of Candidatus Liberibacter asiaticus, the pathogen thought to cause citrus greening disease. The present results indicate that both IGRs tested here should be effective tools for rotation in insecticide-based D. citri management programs. Copyright © 2012 Society of Chemical Industry.

Influence of Tree Size and Application Rate on Expression of Thiamethoxam in Citrus and Its Efficacy Against Diaphorina citri (Hemiptera: Liviidae).

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Langdon, K W; Schumann, R; Stelinski, L L; Rogers, M E

2018-04-02

Neonicotinoids are a key group of insecticides used to manage Diaphorina citri Kuwayama (Hemiptera: Liviidae), in Florida citrus. Diaphorina citri is the vector of Candidatus Liberibacter asiaticus, the presumed causal agent of huanglongbing, a worldwide disease of citrus. A two-season field study was conducted to evaluate the effect of tree size and application rate on the expression of thiamethoxam in young citrus following application to the soil. D. citri adult and nymph abundance was also correlated with thiamethoxam titer in leaves. Tree size and application rate each significantly affected thiamethoxam titer in leaf tissue. The highest mean thiamethoxam titer observed (33.39 ppm) in small trees (mean canopy volume = 0.08 m3) occurred after application of the high rate (0.74 g Platinum 75SG per tree) tested. There was a negative correlation between both nymph and adult abundance with increasing thiamethoxam titer in leaves. A concentration of 64.63 ppm thiamethoxam was required to reach a 1% probability of encountering a flush shoot with at least one adult D. citri, while 19.05 ppm was required for the same probability of encountering nymphs. The LC90 for the field population was 7.62 ppm thiamethoxam when administered through ingestion. Exposure to dosages as low as 7.62 ppm would likely result in sublethal exposure of some proportion of the population, which could exacerbate resistance development. Based on our results, subsequent work should investigate the use of neonicotinoids by foliar rather than soil application to maintain the chemical class in future insecticide management programs in Florida citrus.

Selection of Reference Genes for Expression Studies in Diaphorina citri (Hemiptera: Liviidae).

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Bassan, Meire Menezes; Angelotti-Mendonc A, Je Ssika; Alves, Gustavo Rodrigues; Yamamoto, Pedro Takao; Moura O Filho, Francisco de Assis Alves

2017-12-05

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is considered the main vector of the bacteria associated with huanglongbing, a very serious disease that has threatened the world citrus industry. The absence of efficient control management protocols, including a lack of resistant cultivars, has led to the development of different approaches to study this pathosystem. The production of resistant genotypes relies on D. citri gene expression analyses by RT-qPCR to assess control of the vector population. High-quality, reliable RT-qPCR analyses depend upon proper reference gene selection and validation. However, adequate D. citri reference genes have not yet been identified. In the present study, we evaluated the genes EF 1-α, ACT, GAPDH, RPL7, RPL17, and TUB as candidate reference genes for this insect. Gene expression stability was evaluated using the mathematical algorithms deltaCt, NormFinder, BestKeeper, and geNorm, at five insect developmental stages, grown on two different plant hosts [Citrus sinensis (L.) Osbeck (Sapindales: Rutaceae) and Murraya paniculata (L.) Jack (Sapindales: Rutaceae)]. The final gene ranking was calculated using RefFinder software, and the V-ATPase-A gene was selected for validation. According to our results, two reference genes are recommended when different plant hosts and developmental stages are considered. Considering gene expression studies in D. citri grown on M. paniculata, regardless of the insect developmental stage, GAPDH and RPL7 have the best fit as reference genes in RT-qPCR analyses, whereas GAPDH and EF 1-α are recommended as reference genes in insect studies using C. sinensis. © The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Sulfur volatiles from Allium spp. affect Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), response to citrus volatiles.

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Mann, R S; Rouseff, R L; Smoot, J M; Castle, W S; Stelinski, L L

2011-02-01

The Asian citrus psyllid, Diaphorina citri Kuwayama, vectors Candidatus Liberibacter asiaticus (Las) and Candidatus Liberibacter americanus (Lam), the presumed causal agents of huanglongbing. D. citri generally rely on olfaction and vision for detection of host cues. Plant volatiles from Allium spp. (Alliaceae) are known to repel several arthropod species. We examined the effect of garlic chive (A. tuberosum Rottl.) and wild onion (A. canadense L.) volatiles on D. citri behaviour in a two-port divided T-olfactometer. Citrus leaf volatiles attracted significantly more D. citri adults than clean air. Volatiles from crushed garlic chive leaves, garlic chive essential oil, garlic chive plants, wild onion plants and crushed wild onion leaves all repelled D. citri adults when compared with clean air, with the first two being significantly more repellent than the others. However, when tested with citrus volatiles, only crushed garlic chive leaves and garlic chive essential oil were repellent, and crushed wild onions leaves were not. Analysis of the headspace components of crushed garlic chive leaves and garlic chive essential oil by gas chromatography-mass spectrometry revealed that monosulfides, disulfides and trisulfides were the primary sulfur volatiles present. In general, trisulfides (dimethyl trisulfide) inhibited the response of D. citri to citrus volatiles more than disulfides (dimethyl disulfide, allyl methyl disulfide, allyl disulfide). Monosulfides did not affect the behaviour of D. citri adults. A blend of dimethyl trisulfide and dimethyl disulfide in 1:1 ratio showed an additive effect on inhibition of D. citri response to citrus volatiles. The plant volatiles from Allium spp. did not affect the behaviour of the D. citri ecto-parasitoid Tamarixia radiata (Waterston). Thus, Allium spp. or the tri- and di-sulphides could be integrated into management programmes for D. citri without affecting natural enemies.

Patogenicidade de Beauveria bassiana ao psilídeo Diaphorina citri e compatibilidade do fungo com produtos fitossanitários Beauveria bassiana pathogenicity to Diaphorina citri and compatibility of the fungus with phytosanitary products

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Ana Paula Ferreira Pinto

2012-12-01

Full Text Available O objetivo deste trabalho foi avaliar a patogenicidade de Beauveria bassiana a ninfas de Diaphorina citri (Hemiptera: Psyllidae e verificar a compatibilidade do fungo com produtos fitossanitários e sua persistência em plantas de citros. Ninfas de D. citri foram pulverizadas com B. bassiana, nas concentrações 5x10(6, 1x10(7, 5x10(7, 1x10(8, 5x10(8 e 1x10(9 conídios mL-1, para determinação da concentração letal. Para avaliação da compatibilidade do fungo com produtos fitossanitários, extrato de nim e cinco inseticidas de quatro grupos químicos diferentes foram incorporados individualmente ao meio de cultura BDA em que o fungo foi cultivado. Avaliaram-se o crescimento vegetativo, a esporulação e a viabilidade do entomopatógeno. Plantas de citros, mantidas em casa de vegetação, foram tratadas primeiramente com os produtos fitossanitários e depois com o entomopatógeno. Avaliaram-se os tempos de exposição de 24 horas e de 7 e 14 dias. O fungo foi patogênico às ninfas de D. citri; a CL50 foi de 0,4x10(7 e a CL90 de 6,7x10(7 conidios mL-1, no décimo dia de avaliação. Em laboratório, os produtos fitosssanitários reduzem o crescimento do fungo. Em casa de vegetação, os produtos não afetam a sobrevivência do fungo nas plantas de citros.The objective of this work was to evaluate the pathogenicity of Beauveria bassiana to Diaphorina citri (Hemiptera: Psyllidae nymphs, and to check the compatibility of the fungus with phytosanitary products, and its persistence in citrus plants. Nymphs of D. citri were sprayed with B. bassiana in the concentrations 5x10(6, 1x10(7, 5x10(7, 1x10(8, 5x10(8 and 1x10(9 conidia mL-1 for lethal concentration determination. In order to evaluate the compatibility of the phytosanitary products with the fungus, neem extract and five insecticides of four different chemical groups were individually added to PDA culture medium, in which the fungus was cultivated. Vegetative growth, sporulation and viability

Temperature studies with the Asian citrus psyllid, Diaphorina citri: cold hardiness and temperature thresholds for oviposition.

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Hall, David G; Wenninger, Erik J; Hentz, Matthew G

2011-01-01

This study was conducted to obtain information on the cold hardiness of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), in Florida and to assess upper and lower temperature thresholds for oviposition. The psyllid is an important pest in citrus because it transmits the bacterial pathogens responsible for citrus greening disease, Huanglongbing, considered the most serious citrus disease worldwide. D. citri was first found in Florida during 1998, and the disease was discovered during 2005. Little was known regarding cold hardiness of D. citri, but Florida citrus is occasionally subjected to notable freeze events. Temperature and duration were each significant sources of variation in percent mortality of D. citri subjected to freeze events. Relatively large percentages of adults and nymphs survived after being exposed for several hours to temperatures as low as -5 to -6 °C. Relatively large percentages of eggs hatched after being exposed for several hours to temperatures as low as -8 °C. Research results indicated that adult D. citri become cold acclimated during the winter through exposure to cooler winter temperatures. There was no evidence that eggs became cold acclimated during winter. Cold acclimation in nymphs was not investigated. Research with adult D. citri from laboratory and greenhouse colonies revealed that mild to moderate freeze events were usually nonlethal to the D. citri irrespective of whether they were cold acclimated or not. Upper and lower temperature thresholds for oviposition were investigated because such information may be valuable in explaining the geographic distribution and potential spread of the pest from Florida as well as how cooler winter temperatures might limit population growth. The estimated lower and upper thresholds for oviposition were 16.0 and 41.6 °C, respectively; the estimated temperature of peak oviposition over a 48 h period was 29.6 °C.

Use of the entomopathogenic fungi Metarhizium anisopliae, Cordyceps bassiana and Isaria fumosorosea to control Diaphorina citri (Hemiptera: Psylidae) in Persian lime under field conditions

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The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is a destructive insect pest in the citriculture, because it is an efficient vector of the proteobacteria, ‘Candidatus Liberibacter asiaticus’ (Las), ‘Ca. L. Africanus’ (Laf), and ‘Ca. L. Americanus’ (Lam). These bacteria c...

Analyses of mitogenome sequences revealed that Asian citrus psyllid (Diaphorina citri) from California was related to those from Florida but different from those in Southern China

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Asian citrus psyllid (ACP, Diaphorina citri Kuwayama; Hemiptera: Liviidae) transmits “Candidatus Liberibacter asiaticus” (CLas), an unculturable alpha-proteobacterium associated with citrus Huanglongbing (HLB, yellow shoot disease, also called citrus greening disease). HLB is threatening citrus prod...

[Ovarian development and analysis of mating effects on ovary maturation of Diaphorina citri Kuwayama (Hemiptera: Psyllidae)].

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Dossi, Fábio C A; Cônsoli, Fernando L

2010-01-01

As many other insects with a restricted diet, the citrus psyllid Diaphorina citri Kuwayama, which vector the causing agent of the Huanglongbing, the bacteria Candidatus Liberibacter spp., is intimately associated with symbiotic microorganisms. These mutualist symbionts play a key role on their host nutritional ecology, and are vertically transmitted to the progeny. However, despite the role symbionts play on host development and reproduction, and the growing opportunities of exploitation of the association insect-symbiont to control insect vectored-pathogens, there are very few studies on the host reproductive biology and on the symbiont transovarial transmission. Therefore, we aimed at analyzing the ovary development during D. citri adulthood, and at verifying for the mating requirement as a trigger to initiate ovary development. Newly-emerged D. citri females were grouped as virgin or mated and ovary development was observed during adulthood. Newly-emerged females have immature ovaries, and ovaries remain without any mature eggs until females mate. Once female mates, the vitellogenesis synthesis and uptake are estimulated, and oocytes are quickly developed. Oocytes maturation in ovarioles is metachronic, with only one oocyte developing at a time in each oogenic cycle. Morphological observations of the reproductive system including the ovaries and spermatheca, after the first cluster of eggs is laid, indicated D. citri is polyandrous, and may require multiple mating to develop additional oogenic maturation cycles.

One Target, Two Mechanisms: The Impact of 'Candidatus Liberibacter asiaticus' and Its Vector, Diaphorina citri, on Citrus Leaf Pigments.

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Killiny, Nabil; Nehela, Yasser

2017-07-01

Huanglongbing (HLB) is currently the largest threat to global citrus production. We examined the effect of HLB pathogen 'Candidatus Liberibacter asiaticus' infection or infestation by its vector, Diaphorina citri, on 'Valencia' sweet orange leaf pigments using high-performance liquid chromatography, followed by gene expression analysis for 46 involved genes in carotenoid and chlorophyll biosynthesis pathways. Both 'Ca. L. asiaticus' and D. citri alter the total citrus leaf pigment balance with a greater impact by 'Ca. L. asiaticus'. Although zeaxanthin was accumulated in 'Ca. L. asiaticus'-infected leaves, chlorophyllide a was increased in D. citri-infested plants. Our findings support the idea that both 'Ca. L. asiaticus' and D. citri affect the citrus pigments and promote symptom development but using two different mechanisms. 'Ca. L. asiaticus' promotes chlorophyll degradation but accelerates the biosynthesis of carotenoid pigments, resulting in accumulation of abscisic acid and its precursor, zeaxanthin. Zeaxanthin also has a photoprotective role. By contrast, D. citri induced the degradation of most carotenoids and accelerated chlorophyll biosynthesis, leading to chlorophyllide a accumulation. Chlorophyllide a might have an antiherbivory role. Accordingly, we suggest that citrus plants try to defend themselves against 'Ca. L. asiaticus' or D. citri using multifaceted defense systems, based on the stressor type. These findings will help in better understanding the tritrophic interactions among plant, pathogen, and vector.

Natural parasitism of Diaphorina citri Kuwayama (Hemiptera, Psyllidae nymphs by Tamarixia radiata Waterston (Hymenoptera, Eulophidae in São Paulo orange groves Parasitismo natural de ninfas de Diaphorina citri Kuwayama (Hemiptera, Psyllidae por Tamarixia radiata Waterston (Hymenoptera, Eulophidae em pomares de laranja em São Paulo

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Paulo Eduardo Branco Paiva

2012-12-01

Full Text Available The psyllid Diaphorina citri Kuwayama 1908 has become the main citrus pest species in the state of São Paulo, Brazil, after the introduction of the huanglongbing or citrus greening. This study evaluated the parasitism of 3rd, 4th and 5th instar D. citri nymphs by Tamarixia radiata (Waterston, 1922 in citrus groves under a regimen of regular insecticide applications in ten producing regions: Araraquara, Barretos, Bauru, Botucatu, Franca, Itapetininga, Jaú, Limeira, Lins and São João da Boa Vista. Sixty-nine samples of new branches infested with nymphs of D. citri were collected from 2005 to 2008 in orange groves ranging from 1 to 20 years old, of the varieties Hamlin, Pera, Valencia and Natal. The parasitoid T. radiata is widely distributed in São Paulo orange groves, and was identified in 50 (72% of the samples, showing a mean parasitism rate of 12.4%. The highest parasitism rate was observed in the "summer" (from January through March, with a mean of 25.7%. Nymphal parasitism was above 90% in two samples. The probable causes of the variations in parasitism of D. citri by T. radiata are discussed.O psilídeo Diaphorina citri Kuwayama 1908 tornou-se a principal praga dos citros no estado de São Paulo após a introdução do huanglongbing ou grenning dos citros. Este estudo avaliou as proporções de ninfas de terceiro a quinto ínstares de D. citri parasitadas por Tamarixia radiata (Waterston, 1922 em pomares de laranja submetidos a pulverizações constantes de inseticidas em dez regiões produtoras, Araraquara, Barretos, Bauru, Botucatu, Franca, Itapetininga, Jaú, Limeira, Lins e São João da Boa Vista. Foram coletadas 69 amostras de ramos jovens infestados com ninfas de D. citri obtidas entre 2005 e 2008 em pomares de 1 a 20 anos de idade, das cultivares Hamilin, Pera, Valencia e Natal. O parasitóide T. radiata encontra-se amplamente distribuído em pomares de São Paulo, tendo sido observado em 50 amostras (72%, com taxa média de

Biology of the Huanglongbing vector Diaphorina citri (Hemiptera: Liviidae) on different host plants.

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Alves, G R; Diniz, A J F; Parra, J R P

2014-04-01

Although many studies have been conducted on the development and reproductive potential of Diaphorina citri Kuwayama, 1908 (Hemiptera: Liviidae) in different host species, few have evaluated these parameters on different varieties of the same host species. This study evaluated the influence of five commercial varieties of citrus (Citrus spp. L.)--Hamlin, Natal, Pêra, Ponkan, and Valencia-and orange jasmine [Murraya exotica (L.) Jack] on the development of D. citri. Survival rates for the egg stage were highest on orange jasmine (85.7%) and on Valencia (83.3%). The lowest viability of the nymphal stage was also observed on Hamlin, averaging 57.4%. Values for total viability ranged from 65.9 to 32.6%, and were highest on Valencia. The longest egg-adult development time was on Natal, with a mean of 18.4 d; the shortest total development time was on orange jasmine, with a mean of 17.3 d. Based on the fertility life table, the net reproductive rate (Ro) of D. citri was 2.5 times higher when reared on Valencia than on Hamlin. The other parameters (duration of each generation [T], finite rate of increase [lambda], and innate capacity to increase in number [r(m)]) also demonstrated that Valencia is best suited to this insect. The results obtained for the biological parameters and the fertility life table indicate that Valencia and orange jasmine were the most suitable hosts, whereas Hamlin was least suitable for the development of D. citri. These results provide information for the installation of new citrus groves, especially in the choice of varieties to be planted and the location of different varieties within the groves, with a view toward the management of Huanglongbing or HLB.

Thiamethoxam and imidacloprid drench applications on sweet orange nursery trees disrupt the feeding and settling behaviour of Diaphorina citri (Hemiptera: Liviidae).

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Miranda, Marcelo P; Yamamoto, Pedro T; Garcia, Rafael B; Lopes, João Pa; Lopes, João Rs

2016-09-01

Chemical control is the method most used for management of Diaphorina citri, the vector of the phloem-limited bacteria associated with citrus huanglongbing (HLB) disease. The objectives of this study were to determine the influence of soil-drench applications of neonicotinoids (thiamethoxam and imidacloprid) on the probing behaviour of D. citri on citrus nursery trees, using the electrical penetration graph (EPG) technique, and to measure the D. citri settling behaviour after probing on citrus nursery trees that had received these neonicotinoid treatments. The drench applications of neonicotinoids on citrus nursery trees disrupt D. citri probing, mainly for EPG variables related to phloem sap ingestion, with a significant reduction (≈90%) in the duration of this activity compared with untreated plants in all assessment periods (15, 35 and 90 days after application). Moreover, both insecticides have a repellent effect on D. citri, resulting in significant dispersal of psyllids from treated plants. This study clearly demonstrates the interference of soil-applied neonicotinoids on the feeding and settling behaviour of D. citri on citrus nursery trees, mainly during the phloem ingestion phase. These findings reinforce the recommendation of drench application of neonicotinoids before planting nursery trees as a useful strategy for HLB management. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

Susceptibility of Diaphorina citri (Hemiptera: Liviidae) and Its Parasitoid Tamarixia radiata (Hymenoptera: Eulophidae) to Entomopathogenic Fungi under Laboratory Conditions.

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Ibarra-Cortés, K H; Guzmán-Franco, A W; González-Hernández, H; Ortega-Arenas, L D; Villanueva-Jiménez, J A; Robles-Bermúdez, A

2018-02-01

Diaphorina citri (Kuwayama) is a global pest of citrus that transmits the bacteria associated with the disease, Huanglongbing. Entomopathogenic fungi and the parasitoid Tamarixia radiata (Waterston) are important biological control agents of this pest and likely to interact in D. citri populations. As a basis for interaction studies, we determined the susceptibility of nymphs and adults of D. citri and adults of the parasitoid T. radiata to six fungal isolates from the species Beauveria bassiana s.l. (Bals.-Criv.) Vuill. (isolates B1 and B3), Metarhizium anisopliae s.s. (Metsch.) (Ma129 and Ma65) and Isaria fumosorosea Wize (I2 and Pae). We conducted experiments evaluating infection levels in all three insect groups following inoculation with a series of conidial concentrations (1 × 10 4 -1 × 10 8 conidia mL -1 ). Results showed that D. citri nymphs and T. radiata were more susceptible to fungal isolates than D. citri adults. Overall, B. bassiana and M. anisopliae isolates caused the greatest infection compared with I. fumosorosea isolates in all three groups of insects. Isolates B1 (B. bassiana) and Ma129 (M. anisopliae) infected a greater proportion of adults and nymphs of D. citri, respectively. Both isolates of B. bassiana caused greater infection in T. radiata compared with isolates of the other fungal species. We propose that isolates B1 and Ma129 are the strongest candidates for control of D. citri. Our results represent the first report of entomopathogenic fungi infecting T. radiata, and the basis for future studies to design a biological control programme that uses both agents more efficiently against D. citri populations.

The Influence of Learning on Host Plant Preference in a Significant Phytopathogen Vector, Diaphorina citri.

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Dara G Stockton

Full Text Available Although specialist herbivorous insects are guided by innate responses to host plant cues, host plant preference may be influenced by experience and is not dictated by instinct alone. The effect of learning on host plant preference was examined in the Asian citrus psyllid, Diaphorina citri; vector of the causal agent of citrus greening disease or huanglongbing. We investigated: a whether development on specific host plant species influenced host plant preference in mature D. citri; and b the extent of associative learning in D. citri in the form of simple and compound conditioning. Learning was measured by cue selection in a 2-choice behavioral assay and compared to naïve controls. Our results showed that learned responses in D. citri are complex and diverse. The developmental host plant species influenced adult host plant preference, with female psyllids preferring the species on which they were reared. However, such preferences were subject to change with the introduction of an alternative host plant within 24-48 hrs, indicating a large degree of experience-dependent response plasticity. Additionally, learning occurred for multiple sensory modalities where novel olfactory and visual environmental cues were associated with the host plant. However, males and females displayed differing discriminatory abilities. In compound conditioning tasks, males exhibited recognition of a compound stimulus alone while females were capable of learning the individual components. These findings suggest D. citri are dynamic animals that demonstrate host plant preference based on developmental and adult experience and can learn to recognize olfactory and visual host plant stimuli in ways that may be sex specific. These experience-based associations are likely used by adults to locate and select suitable host plants for feeding and reproduction and may suggest the need for more tailored lures and traps, which reflect region-specific cultivars or predominate

The Influence of Learning on Host Plant Preference in a Significant Phytopathogen Vector, Diaphorina citri.

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Stockton, Dara G; Martini, Xavier; Patt, Joseph M; Stelinski, Lukasz L

2016-01-01

Although specialist herbivorous insects are guided by innate responses to host plant cues, host plant preference may be influenced by experience and is not dictated by instinct alone. The effect of learning on host plant preference was examined in the Asian citrus psyllid, Diaphorina citri; vector of the causal agent of citrus greening disease or huanglongbing. We investigated: a) whether development on specific host plant species influenced host plant preference in mature D. citri; and b) the extent of associative learning in D. citri in the form of simple and compound conditioning. Learning was measured by cue selection in a 2-choice behavioral assay and compared to naïve controls. Our results showed that learned responses in D. citri are complex and diverse. The developmental host plant species influenced adult host plant preference, with female psyllids preferring the species on which they were reared. However, such preferences were subject to change with the introduction of an alternative host plant within 24-48 hrs, indicating a large degree of experience-dependent response plasticity. Additionally, learning occurred for multiple sensory modalities where novel olfactory and visual environmental cues were associated with the host plant. However, males and females displayed differing discriminatory abilities. In compound conditioning tasks, males exhibited recognition of a compound stimulus alone while females were capable of learning the individual components. These findings suggest D. citri are dynamic animals that demonstrate host plant preference based on developmental and adult experience and can learn to recognize olfactory and visual host plant stimuli in ways that may be sex specific. These experience-based associations are likely used by adults to locate and select suitable host plants for feeding and reproduction and may suggest the need for more tailored lures and traps, which reflect region-specific cultivars or predominate Rutaceae in the area

β-caryophyllene emitted from a transgenic Arabidopsis or chemical dispenser repels Diaphorina citri, vector of Candidatus Liberibacters.

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Alquézar, Berta; Volpe, Haroldo Xavier Linhares; Magnani, Rodrigo Facchini; de Miranda, Marcelo Pedreira; Santos, Mateus Almeida; Wulff, Nelson Arno; Bento, Jose Mauricio Simões; Parra, José Roberto Postali; Bouwmeester, Harro; Peña, Leandro

2017-07-17

Production of citrus, the main fruit tree crop worldwide, is severely threatened by Huanglongbing (HLB), for which as yet a cure is not available. Spread of this bacterial disease in America and Asia is intimately connected with dispersal and feeding of the insect vector Diaphorina citri, oligophagous on rutaceous host plants. Effective control of this psyllid is an important component in successful HLB management programs. Volatiles released from the non-host guava have been shown to be repellent to the psyllid and to inhibit its response to citrus odour. By analysing VOC emission from guava we identified one volatile compound, (E)-β-caryophyllene, which at certain doses exerts a repellent effect on D. citri. Non-host plant rejection mediated by (E)-β-caryophyllene is demonstrated here by using Arabidopsis over-expression and knock-out lines. For the first time, results indicate that genetically engineered Arabidopsis plants with modified emission of VOCs can alter the behaviour of D. citri. This study shows that transgenic plants with an inherent ability to release (E)-β-caryophyllene can potentially be used in new protection strategies of citrus trees against HLB.

Host Range Testing of Diaphorencyrtus aligarhensis (Hymenoptera: Encyrtidae) for Use in Classical Biological Control of Diaphorina citri (Hemiptera: Liviidae) in California.

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Bistline-East, Allison; Pandey, Raju; Kececi, Mehmet; Hoddle, Mark S

2015-06-01

Host range tests for Diaphorencyrtus aligarhensis (Shafee, Alam, & Agarwal) (Hymenoptera: Encyrtidae), an endoparasitoid of Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), sourced from Punjab Pakistan, were conducted in quarantine at the University of California, Riverside, CA. Seven nontarget psyllid species representing four psyllid families were exposed to mated D. aligarhensis females in four different treatment types: 1) short sequential no-choice treatments, 2) prolonged sequential no-choice treatments, 3) prolonged no-choice static treatments, and 4) choice treatments. Selection of nontarget psyllid species was based on phylogenetic proximity to D. citri, likelihood of being encountered by D. aligarhensis in the prospective release areas in California, and psyllid species in biological control of invasive weeds. D. aligarhensis exhibited high host affinity to D. citri, and only parasitized one nontarget species, the pestiferous potato psyllid, Bactericera cockerelli (Sulc), at low levels (citri. Results presented here suggest D. aligarhensis poses minimal risk to nontarget psyllid species in California. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

GENETIC Diaphorina citri DIVERSITY ON CITRUS CROPS OF THE VALLE DEL CAUCA AND QUINDÍO (COLOMBIA

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MIGUEL ANGEL MONCAYO-DONOSO

2014-07-01

Full Text Available The Asiatic psyllid Diaphorina citri (Hemiptera: Psyllidae is the main vector of Candidatus liberibacter, which causes the Huanglongbing HLB disease, known for devastating citrus in the world but not yet reported in Colombia. The genetic variability of the D. citri population was studied through sequencing the COI mitochondrial gene as molecular marker. Adults were collected in citrus producing zones of the Colombian Valle del Cauca and Quindío. Amplification was performed with two pairs of specific primers for Hemiptera. The PCR products were sequenced at Macrogen-Korea, obtaining a total of 124 sequences. For the bioinformatic analysis, the Vector NTI 11.5, Harlequin V 3.5, MEGA 5 and MAFFT 6 programs were used. The molecular diversity indices between populations were similar, revealing a common origin and a recent split of the populations excluding a significant genetic differentiation associated to variations of the bacterium, however the haplotype diversity index was higher than the nucleotide diversity index. The latter one showed a low number of polymorphic sites, indicating that the D. citri populations are expanding. The study of the vector’s genetic variability is a tool for the prediction of likely scenarios for the spread of diseases.

Evaluation of six different groups of insecticides for the control of citrus psylla Diaphorina citri (Hemiptera: Psyllidae

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Rakhmin Gul

2005-01-01

Full Text Available Studies on the efficacy of different insecticides against citrus psylla, Diaphorina citri (Hemiptera: Psyllidae were carried out at Agricultural Research Institute, Tarnab, Peshawar, Pakistan. Six insecticides viz. Actara 25 WG, (thiamethoxam Cascade 10 DC (Flufenoxuron, Match 050 EC (lufenuron, Thiodan 35 EC (endosulfan, Karate 2.5 EC (α-cyhalothrin, and Supracide 40 EC (methidathion, were tested for their effectiveness against D. citri. After first spray overall mean population of D. citri was 3.63, 4.75, 5.59, 6.66, 7.47, 8.11 per six inches tender shoot on Actara 25 WG, Cascade 10 DC, Match 050 EC, Thiodan 35 EC, Karate 2.5 EC and Supracide 40 EC treated plants respectively, while on control plants the population was 12.39. Similarly, after the second spray of each of the same insecticides the population of D. citri was 2.65, 4.23, 5.61, 6.41, 7.35 and 8.73 respectively. Where in controls there were 15.18 psyllids. Percent decrease of D. citri population in comparison to control after the first spray was highest in Actara 25 WG (72.20 followed by Cascade 10 DC (62.91, Match 050 EC (54.07, Thiodan 35 EC (47.61, Karate 2.5 EC (38.94 and Supracide 40 EC (35.74. After the second spray percent decrease over control recorded was highest in Actara 25 WG (83.54, followed by Cascade 10 DC (71.08, Match 050 EC (63.94, Thiodan 35 EC (60.79, Karate 2.5 EC (52.52 and Supracide 40 EC (45.62.

Geographic distribution of habitat, development, and population growth rates of the Asian citrus psyllid, Diaphorina citri, in Mexico.

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López-Collado, José; Isabel López-Arroyo, J; Robles-García, Pedro L; Márquez-Santos, Magdalena

2013-01-01

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is an introduced pest in Mexico and a vector of huanglongbing, a lethal citrus disease. Estimations of the habitat distribution and population growth rates of D. citri are required to establish regional and areawide management strategies and can be used as a pest risk analysis tools. In this study, the habitat distribution of D. citri in Mexico was computed with MaxEnt, an inductive, machine-learning program that uses bioclimatic layers and point location data. Geographic distributions of development and population growth rates were determined by fitting a temperature-dependent, nonlinear model and projecting the rates over the target area, using the annual mean temperature as the predictor variable. The results showed that the most suitable regions for habitat of D. citri comprise the Gulf of Mexico states, Yucatán Peninsula, and areas scattered throughout the Pacific coastal states. Less suitable areas occurred in northern and central states. The most important predictor variables were related to temperature. Development and growth rates had a distribution wider than habitat, reaching some of the northern states of México. Habitat, development, and population growth rates were correlated to each other and with the citrus producing area. These relationships indicated that citrus producing states are within the most suitable regions for the occurrence, development, and population growth of D. citri, therefore increasing the risk of huanglongbing dispersion.

Effects of different temperature regimes on survival of Diaphorina citri and its endosymbiotic bacterial communities.

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Hussain, Mubasher; Akutse, Komivi Senyo; Ravindran, Keppanan; Lin, Yongwen; Bamisile, Bamisope Steve; Qasim, Muhammad; Dash, Chandra Kanta; Wang, Liande

2017-09-01

The Asian citrus psyllid, Diaphorina citri, is a major pest of citrus and vector of citrus greening (huanglongbing) in Asian. In our field-collected psyllid samples, we discovered that Fuzhou (China) and Faisalabad (Pakistan), populations harbored an obligate primary endosymbiont Candidatus Carsonella (gen. nov.) with a single species, Candidatus Carsonella ruddii (sp. nov.) and a secondary endosymbiont, Wolbachia surface proteins (WSP) which are intracellular endosymbionts residing in the bacteriomes. Responses of these symbionts to different temperatures were examined and their host survival assessed. Diagnostic PCR assays showed that the endosymbionts infection rates were not significantly reduced in both D. citri populations after 24 h exposure to cold or heat treatments. Although quantitative PCR assays showed significant reduction of WSP relative densities at 40°C for 24 h, a substantial decrease occurred as the exposure duration increased beyond 3 days. Under the same temperature regimes, Ca. C. ruddii density was initially less affected during the first exposure day, but rapidly reduced at 3-5 days compared to WSP. However, the mortality of the psyllids increased rapidly as exposure time to heat treatment increased. The responses of the two symbionts to unfavorable temperature regimes highlight the complex host-symbionts interactions between D. citri and its associated endosymbionts. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

Latency and Persistence of 'Candidatus Liberibacter asiaticus' in Its Psyllid Vector, Diaphorina citri (Hemiptera: Liviidae).

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Canale, Maria Cristina; Tomaseto, Arthur Fernando; Haddad, Marineia de Lara; Della Coletta-Filho, Helvécio; Lopes, João Roberto Spotti

2017-03-01

Although 'Candidatus Liberibacter asiaticus' (Las) is a major pathogen associated with citrus huanglongbing (HLB), some characteristics of transmission by the psyllid vector Diaphorina citri are not fully understood. We examined the latent period and persistence of transmission of Las by D. citri in a series of experiments at 25°C, in which third-instar psyllid nymphs and 1-week-old adults were confined on infected citrus for an acquisition access period (AAP), and submitted to sequential inoculation access periods (IAPs) on healthy citrus seedlings. The median latent period (LP 50 , i.e., acquisition time after which 50% of the individuals can inoculate) of 16.8 and 17.8 days for psyllids that acquired Las as nymphs and adults, respectively, was determined by transferring single individuals in 48-h IAPs. Inoculation events were intermittent and randomly distributed over the IAPs, but were more frequent after acquisition by nymphs. A minimum latent period of 7 to 10 days was observed by transferring groups of 10 psyllids in 48-h IAPs, after a 96-h AAP by nymphs. Psyllids transmitted for up to 5 weeks, when submitted to sequential 1-week IAPs after a 14-day AAP as nymphs. The long latent period and persistence of transmission are indirect evidences of circulative propagation of Las in D. citri.

Insecticidal suppression of Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae vector of huanglongbing pathogens.

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Jawwad A Qureshi

Full Text Available Diaphorina citri vectors pathogens that cause 'huanglongbing' or citrus greening disease which poses a serious threat to citrus production worldwide. Vector suppression is critical to reduce disease spread. Efficacy is a main concern when choosing an insecticide. Insecticidal treatments of 49 products or 44 active ingredients (a.i labeled or experimental were field tested between 2005-2013 as foliar sprays (250 treatments, 39 a.i or soil applications (47 treatments, 9 a.i to control D. citri in citrus. A combined effect of nymphal and adult suppression in response to sprays of 23 insecticides representing 9 modes of action (MoA groups and 3 unknown MoA provided more than 90% reduction of adult D. citri over 24-68 days. Observable effects on nymphs were generally of shorter duration due to rapid maturation of flush. However, reduction of 76-100% nymphs or adults over 99-296 days was seen on young trees receiving drenches of the neonicotinoids imidacloprid, thiamethoxam or clothianidin (MoA 4A and a novel anthranilic diamide, cyantraniliprole (MoA 28. Effective products identified for foliar sprays to control D. citri provide sufficient MoA groups for rotation to delay evolution of insecticide resistance by D. citri and other pests. However, cyantraniliprole is now the only available alternative for rotation with neonicotinoids in soil application to young trees. Sprays of up to eight of the most effective insecticides could be rotated over a year without repetition of any MoA and little or no recourse to neonicotinoids or cyantraniliprole, so important for protection of young trees. Other considerations effecting decisions of what and when to spray include prevalence of huanglongbing, pest pressure, pre-harvest intervals, overall budget, equipment availability, and conservation of beneficial arthropods. Examples of spray programs utilizing broad-spectrum and relatively selective insecticides are provided to improve vector management and may vary

Insecticidal suppression of Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae) vector of huanglongbing pathogens.

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Qureshi, Jawwad A; Kostyk, Barry C; Stansly, Philip A

2014-01-01

Diaphorina citri vectors pathogens that cause 'huanglongbing' or citrus greening disease which poses a serious threat to citrus production worldwide. Vector suppression is critical to reduce disease spread. Efficacy is a main concern when choosing an insecticide. Insecticidal treatments of 49 products or 44 active ingredients (a.i) labeled or experimental were field tested between 2005-2013 as foliar sprays (250 treatments, 39 a.i) or soil applications (47 treatments, 9 a.i) to control D. citri in citrus. A combined effect of nymphal and adult suppression in response to sprays of 23 insecticides representing 9 modes of action (MoA) groups and 3 unknown MoA provided more than 90% reduction of adult D. citri over 24-68 days. Observable effects on nymphs were generally of shorter duration due to rapid maturation of flush. However, reduction of 76-100% nymphs or adults over 99-296 days was seen on young trees receiving drenches of the neonicotinoids imidacloprid, thiamethoxam or clothianidin (MoA 4A) and a novel anthranilic diamide, cyantraniliprole (MoA 28). Effective products identified for foliar sprays to control D. citri provide sufficient MoA groups for rotation to delay evolution of insecticide resistance by D. citri and other pests. However, cyantraniliprole is now the only available alternative for rotation with neonicotinoids in soil application to young trees. Sprays of up to eight of the most effective insecticides could be rotated over a year without repetition of any MoA and little or no recourse to neonicotinoids or cyantraniliprole, so important for protection of young trees. Other considerations effecting decisions of what and when to spray include prevalence of huanglongbing, pest pressure, pre-harvest intervals, overall budget, equipment availability, and conservation of beneficial arthropods. Examples of spray programs utilizing broad-spectrum and relatively selective insecticides are provided to improve vector management and may vary depending on

Sperm morphology of the leafhopper Diaphorina citri Kuwayama (Hemiptera: Sternorrhyncha: Psylloidea: Liviidae).

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Barcellos, Marcelo Silva; Cossolin, Jamile Fernanda Silva; Dias, Glenda; Lino-Neto, José

2017-08-01

In this study, by using light and electron microscopy, we describe the sperm morphology of the leafhopper Diaphorina citri, a serious pest of citrus throughout the world. In this species the sperm measures 538.49±8.75μm in length, and as observed in psylloids, the sperm, when manipulated, opens into two filaments, one of which is attached to the nuclear base and the other becomes free. Along the flagellum, and only of it, there are lateral projections, about 2μm in length. Furthermore, at the end of the flagellum three appendages, with approximately 7μm in length, are observed. The head region is formed by the nucleus with compact chromatin, and, parallel to it, a structure of median electron density that extends about 25μm in length ahead of the nuclear tip. The flagellum consists of an axoneme with a 9+9+2 microtubule arrangement, two mitochondrial derivatives, and two accessory bodies each with two regions of different electron density. The presence of lateral projections is a characteristic observed in other Sternorrhyncha. As seen previously, the sperm opening in two filaments, when manipulated, was observed only in Psylloidea, and the presence of the three appendices at the end of the flagellum distinguishes D. citri from the other Psylloidea species studied. Copyright © 2017 Elsevier Ltd. All rights reserved.

RNA interference of carboxyesterases causes nymph mortality in the Asian citrus psyllid, Diaphorina citri.

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Kishk, Abdelaziz; Anber, Helmy A I; AbdEl-Raof, Tsamoh K; El-Sherbeni, AbdEl-Hakeem D; Hamed, Sobhy; Gowda, Siddarame; Killiny, Nabil

2017-03-01

Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is an important pest of citrus. In addition, D. citri is the vector of Huanglongbing, a destructive disease in citrus, also known as citrus greening disease caused by Candidatus Liberibacter asiaticus. Huanglongbing causes huge losses for citrus industries. Insecticide application for D. citri is the major strategy to prevent disease spread. The heavy use of insecticides causes development of insecticide resistance. We used RNA interference (RNAi) to silence genes implicated in pesticide resistance in order to increase the susceptibility. The activity of dsRNA to reduce the expression of carboxyesterases including esterases FE4 (EstFE4) and acetylcholinesterases (AChe) in D. citri was investigated. The dsRNA was applied topically to the fourth and fifth instars of nymphs. We targeted several EstFE4 and AChe genes using dsRNA against a consensus sequence for each of them. Five concentrations (25, 50, 75, 100, 125 ng/μl) from both dsRNAs were used. The treatments with the dsRNA caused concentration dependent nymph mortality. The highest gene expression levels of both AChe and EstFE4 were found in the fourth and fifth nymphal instars. Gene expression analysis showed that AChe genes were downregulated in emerged adults from dsRNA-AChe-treated nymphs compared to controls. However, EstFE4 genes were not affected. In the same manner, treatment with dsRNA-EstFE4 reduced expression level of EstFE4 genes in emerged adults from treated nymphs, but did not affect the expression of AChe genes. In the era of environmentally friendly control strategies, RNAi is a new promising venue to reduce pesticide applications. © 2017 Wiley Periodicals, Inc.

Spatial distribution of Diaphorina citri Kuwayama (Hemiptera: Psyllidae in citrus orchards Distribuição espacial de Diaphorina citri Kuwayama (Hemiptera: Psyllidae em pomares de citros

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Marilia Gregolin Costa

2010-10-01

Full Text Available The psyllid Diaphorina citri Kuwayama is one of the most important pests of citrus, mainly because it is the vector of the bacterium that causes huanglongbing (HLB or 'Greening' disease. To study the spatial distribution of nymphs and adults of this pest, an experiment was carried out in two 'Valencia' sweet orange orchards, four and 12 years of age, established in Matão, central area of São Paulo state, Brazil. The following dispersion indices were used to study pest aggregation in the citrus plants: variance/mean relationship (I, index of Morisita (Iδ, coefficient of Green (Cx, the exponent of negative binomial distribution, common k (c and Taylor's Power Law for each sampling. The negative binomial distribution was more representative of the spatial distribution of this psyllid, for both nymphs and adults. For most samplings, psyllid nymphs found in branches and adults caught in traps had an aggregated distribution.O psilídeo Diaphorina citri Kuwayama tornou-se nos últimos anos uma das mais importantes pragas na cultura de citros, principalmente pelos prejuízos causados às plantas por ser o transmissor da bactéria causadora da doença Huanglongbing (HLB ou 'Greening'. Com a finalidade de estudar a distribuição espacial de ninfas e adultos desta praga, instalaram-se experimentos em duas áreas de citros com histórico de ocorrência de HLB, no município de Matão (região central do Estado de São Paulo, em plantas de laranja 'Valência', com quatro e 12 anos de idade. Para estudo da agregação da população nas plantas, foram utilizados os seguintes índices de dispersão: razão variância/média (I, índice de Morisita (Iδ, coeficiente de Green (Cx e expoente da distribuição binomial negativa, k comum (c e lei da potência de Taylor para cada amostragem. A distribuição binomial negativa foi o modelo mais adequado para representar a distribuição espacial do psilídeo, tanto para ninfas como para adultos. Na maioria das

Genetic variation and potential coinfection of Wolbachia among widespread Asian citrus psyllid (Diaphorina citri Kuwayama) populations.

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Chu, Chia-Ching; Hoffmann, Mark; Braswell, W Evan; Pelz-Stelinski, Kirsten S

2017-12-29

Wolbachia can profoundly influence the survival, reproduction, and defenses of insect hosts. These interactions could potentially be harnessed for managing pests or insect-transmitted diseases. Diaphorina citri Kuwayama is a phloem-feeding pest capable of transmitting the putative causal agent of citrus greening, Candidatus Liberibacter asiaticus (CLas). Like many insects, D. citri is also infected with Wolbachia (wDi). Recent studies indicate that the relative abundance of wDi could be associated with the abundance of CLas, and that wDi may contribute to regulating expression of phage lytic cycle genes in CLas, suggesting the need for better understanding of wDi biology in general. This study investigated the genetic diversity of wDi among D. citri in populations spanning eleven countries and two U.S. territories. Six Wolbachia genes, wsp, coxA, fbpA, ftsZ, gatB, and hcpA, were sequenced and compared across samples. Two prevalent wDi strains were identified across the samples, and screening of clone libraries revealed possible coinfection of wDi strains in specific populations. D. citri mitochondrial cytochrome oxidase subunit I gene (mtCOI) were more divergent between D. citri populations that were infected with different wDi strains or had different infection statuses (single infection vs. coinfection). While we could not eliminate the possibility that maternal transmission may contribute to such patterns, it is also possible that wDi may induce cytoplasmic incompatibility in their host. These findings should contribute to the understanding of wDi population ecology, which may facilitate manipulation of this endosymbiont for management of citrus greening disease worldwide. © 2017 The Authors. Insect Science published by John Wiley & Sons Australia, Ltd on behalf of Institute of Zoology, Chinese Academy of Sciences.

Phylogeography of Diaphorina citri (Hemiptera: Liviidae) and its primary endosymbiont, 'Candidatus Carsonella ruddii': an evolutionary approach to host-endosymbiont interaction.

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Wang, Yanjing; Lu, Jinming; Beattie, George Ac; Islam, Mohammad R; Om, Namgay; Dao, Hang T; Van Nguyen, Liem; Zaka, Syed M; Guo, Jun; Tian, Mingyi; Deng, Xiaoling; Tan, Shunyun; Holford, Paul; He, Yurong; Cen, Yijing

2018-03-25

In insects, little is known about the co-evolution between their primary endosymbionts and hosts at the intraspecific level. This study examined co-diversification between the notorious agricultural pest Diaphorina citri and its primary endosymbionts (P-endosymbiont), 'Candidatus Carsonella ruddii' at the population level. Maximum likelihood, haplotype network, principal components and Bayesian clustering identified three lineages for D. citri and its P-endosymbiont: a Western clade containing individuals from Pakistan, Bhutan (Phuentsholing), Vietnam (Son La), USA, Myanmar and China (Ruili, Yunnan); a Central clade, with accessions originating from Southwest China, Bhutan (Tsirang) and Bangladesh; and an Eastern clade containing individuals from Southeast Asia, and East and South China. A more diverse genetic structure was apparent in the host mitochondrial DNA than their P-endosymbionts; however, the two sets of data were strongly congruent. This study provides evidence for the co-diversification of D. citri and its P-endosymbiont during the migration from South Asia to East and Southeast Asia. We also suggest that the P-endosymbiont may facilitate investigations into the genealogy and migration history of the host. The biogeography of D. citri and its P-endosymbiont indicated that D. citri colonized and underwent a secondary dispersal from South Asia to East and Southeast Asia. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

Natural parasitism of Diaphorina citri Kuwayama (Hemiptera, Psyllidae nymphs by Tamarixia radiata Waterston (Hymenoptera, Eulophidae in São Paulo orange groves

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Paulo Eduardo Branco Paiva

2012-12-01

Full Text Available The psyllid Diaphorina citri Kuwayama 1908 has become the main citrus pest species in the state of São Paulo, Brazil, after the introduction of the huanglongbing or citrus greening. This study evaluated the parasitism of 3rd, 4th and 5th instar D. citri nymphs by Tamarixia radiata (Waterston, 1922 in citrus groves under a regimen of regular insecticide applications in ten producing regions: Araraquara, Barretos, Bauru, Botucatu, Franca, Itapetininga, Jaú, Limeira, Lins and São João da Boa Vista. Sixty-nine samples of new branches infested with nymphs of D. citri were collected from 2005 to 2008 in orange groves ranging from 1 to 20 years old, of the varieties Hamlin, Pera, Valencia and Natal. The parasitoid T. radiata is widely distributed in São Paulo orange groves, and was identified in 50 (72% of the samples, showing a mean parasitism rate of 12.4%. The highest parasitism rate was observed in the "summer" (from January through March, with a mean of 25.7%. Nymphal parasitism was above 90% in two samples. The probable causes of the variations in parasitism of D. citri by T. radiata are discussed.

Neonicotinoid-Induced Mortality of Diaphorina Citri (Hemiptera: Liviidae) is Affected by Route of Exposure.

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Langdon, Kevin W; Rogers, Michael E

2017-10-01

The use of neonicotinoids in citrus (Rutaceae) has increased substantially to help manage the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), a vector of the devastating citrus disease, huanglongbing (HLB). In citrus pest management programs, neonicotinoids are most often applied to the soil as a drench and move through xylem channels from the roots into the foliage. We developed a novel assay to quantify the dose required to kill D. citri following ingestion and compare it with the dose required to kill by contact. The LC50 of the laboratory strain for ingestion of imidacloprid, thiamethoxam, and clothianidin were each approximately 10-fold greater than the respective LC50 by contact exposure. Four field populations were tested to validate comparative exposure of the laboratory strain to imidacloprid and determine the relative susceptibility of field populations to imidacloprid by exposure through ingestion and contact. The contact assay exhibited low (10) RR50 values were observed for the Lake Placid and Lake Alfred populations using the contact and the ingestion method. This research demonstrates that the ingestion assay method described herein is more sensitive in detection of low-level resistance and should be the standard methodology used in monitoring for resistance to systemic insecticides for this global pest. We found D. citri populations with a lower than expected susceptibility to neonicotinoids in the field, which warrants the implementation of resistance management practices to preserve the utility of soil-applied neonicotinoids in citrus. © The Author 2017. Published by Oxford University Press on behalf of Entomological Society of America.

Metabolomic Response to Huanglongbing: Role of Carboxylic Compounds in Citrus sinensis Response to 'Candidatus Liberibacter asiaticus' and Its Vector, Diaphorina citri.

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Killiny, Nabil; Nehela, Yasser

2017-08-01

Huanglongbing, a destructive disease of citrus, is caused by the fastidious bacterium 'Candidatus Liberibacter asiaticus' and transmitted by Asian citrus psyllid, Diaphorina citri. The impact of 'Ca. L. asiaticus' infection or D. citri infestation on Valencia sweet orange (Citrus sinensis) leaf metabolites was investigated using gas chromatography mass spectrometry, followed by gene expression analysis for 37 genes involved in jasmonic acid (JA), salicylic acid (SA), and proline-glutamine pathways. The total amino acid abundance increased after 'Ca. L. asiaticus' infection, while the total fatty acids increased dramatically after infestation with D. citri, compared with control plants. Seven amino acids (glycine, l-isoleucine, l-phenylalanine, l-proline, l-serine, l-threonine, and l-tryptophan) and five organic acids (benzoic acid, citric acid, fumaric acid, SA, and succinic acid) increased in 'Ca. L. asiaticus'-infected plants. On the other hand, the abundance of trans-JA and its precursor α-linolenic increased in D. citri-infested plants. Surprisingly, the double attack of both D. citri infestation and 'Ca. L. asiaticus' infection moderated the metabolic changes in all chemical classes studied. In addition, the gene expression analysis supported these results. Based on these findings, we suggest that, although amino acids such as phenylalanine are involved in citrus defense against 'Ca. L. asiaticus' infection through the activation of an SA-mediated pathway, fatty acids, especially α-linolenic acid, are involved in defense against D. citri infestation via the induction of a JA-mediated pathway.

Protein interaction networks at the host-microbe interface in Diaphorina citri, the insect vector of the citrus greening pathogen.

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Ramsey, J S; Chavez, J D; Johnson, R; Hosseinzadeh, S; Mahoney, J E; Mohr, J P; Robison, F; Zhong, X; Hall, D G; MacCoss, M; Bruce, J; Cilia, M

2017-02-01

The Asian citrus psyllid ( Diaphorina citri) is the insect vector responsible for the worldwide spread of ' Candidatus Liberibacter asiaticus' (CLas), the bacterial pathogen associated with citrus greening disease. Developmental changes in the insect vector impact pathogen transmission, such that D. citri transmission of CLas is more efficient when bacteria are acquired by nymphs when compared with adults. We hypothesize that expression changes in the D. citri immune system and commensal microbiota occur during development and regulate vector competency. In support of this hypothesis, more proteins, with greater fold changes, were differentially expressed in response to CLas in adults when compared with nymphs, including insect proteins involved in bacterial adhesion and immunity. Compared with nymphs, adult insects had a higher titre of CLas and the bacterial endosymbionts Wolbachia, Profftella and Carsonella. All Wolbachia and Profftella proteins differentially expressed between nymphs and adults are upregulated in adults, while most differentially expressed Carsonella proteins are upregulated in nymphs. Discovery of protein interaction networks has broad applicability to the study of host-microbe relationships. Using protein interaction reporter technology, a D. citri haemocyanin protein highly upregulated in response to CLas was found to physically interact with the CLas coenzyme A (CoA) biosynthesis enzyme phosphopantothenoylcysteine synthetase/decarboxylase. CLas pantothenate kinase, which catalyses the rate-limiting step of CoA biosynthesis, was found to interact with a D. citri myosin protein. Two Carsonella enzymes involved in histidine and tryptophan biosynthesis were found to physically interact with D. citri proteins. These co-evolved protein interaction networks at the host-microbe interface are highly specific targets for controlling the insect vector responsible for the spread of citrus greening.

Double-stranded RNA uptake through topical application, mediates silencing of five CYP4 genes and suppresses insecticide resistance in Diaphorina citri.

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Killiny, Nabil; Hajeri, Subhas; Tiwari, Siddharth; Gowda, Siddarame; Stelinski, Lukasz L

2014-01-01

Silencing of genes through RNA interference (RNAi) in insects has gained momentum during the past few years. RNAi has been used to cause insect mortality, inhibit insect growth, increase insecticide susceptibility, and prevent the development of insecticide resistance. We investigated the efficacy of topically applied dsRNA to induce RNAi for five Cytochrome P450 genes family 4 (CYP4) in Diaphorina citri. We previously reported that these CYP4 genes are associated with the development of insecticide resistance in D. citri. We targeted five CYP4 genes that share a consensus sequence with one dsRNA construct. Quantitative PCR confirmed suppressed expression of the five CYP4 genes as a result of dsRNA topically applied to the thoracic region of D. citri when compared to the expression levels in a control group. Western blot analysis indicated a reduced signal of cytochrome P450 proteins (45 kDa) in adult D. citri treated with the dsRNA. In addition, oxidase activity and insecticide resistance were reduced for D. citri treated with dsRNA that targeted specific CYP4 genes. Mortality was significantly higher in adults treated with dsRNA than in adults treated with water. Our results indicate that topically applied dsRNA can penetrate the cuticle of D. citri and induce RNAi. These results broaden the scope of RNAi as a mechanism to manage pests by targeting a broad range of genes. The results also support the application of RNAi as a viable tool to overcome insecticide resistance development in D. citri populations. However, further research is needed to develop grower-friendly delivery systems for the application of dsRNA under field conditions. Considering the high specificity of dsRNA, this tool can also be used for management of D. citri by targeting physiologically critical genes involved in growth and development.

Double-stranded RNA uptake through topical application, mediates silencing of five CYP4 genes and suppresses insecticide resistance in Diaphorina citri.

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Nabil Killiny

Full Text Available Silencing of genes through RNA interference (RNAi in insects has gained momentum during the past few years. RNAi has been used to cause insect mortality, inhibit insect growth, increase insecticide susceptibility, and prevent the development of insecticide resistance. We investigated the efficacy of topically applied dsRNA to induce RNAi for five Cytochrome P450 genes family 4 (CYP4 in Diaphorina citri. We previously reported that these CYP4 genes are associated with the development of insecticide resistance in D. citri. We targeted five CYP4 genes that share a consensus sequence with one dsRNA construct. Quantitative PCR confirmed suppressed expression of the five CYP4 genes as a result of dsRNA topically applied to the thoracic region of D. citri when compared to the expression levels in a control group. Western blot analysis indicated a reduced signal of cytochrome P450 proteins (45 kDa in adult D. citri treated with the dsRNA. In addition, oxidase activity and insecticide resistance were reduced for D. citri treated with dsRNA that targeted specific CYP4 genes. Mortality was significantly higher in adults treated with dsRNA than in adults treated with water. Our results indicate that topically applied dsRNA can penetrate the cuticle of D. citri and induce RNAi. These results broaden the scope of RNAi as a mechanism to manage pests by targeting a broad range of genes. The results also support the application of RNAi as a viable tool to overcome insecticide resistance development in D. citri populations. However, further research is needed to develop grower-friendly delivery systems for the application of dsRNA under field conditions. Considering the high specificity of dsRNA, this tool can also be used for management of D. citri by targeting physiologically critical genes involved in growth and development.

Effects of methoprene, a juvenile hormone analog, on survival of various developmental stages, adult emergence, reproduction and behavior of Asian citrus psyllid, Diaphorina citri Kuwayama.

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Brar, Gurpreet S; Meyer, Wendy; Stelinski, Lukasz L

2015-12-01

The Asian citrus citrus psyllid, Diaphorina citri Kuwayama, transmits a bacterium that causes huanglongbing in citrus. Frequent and repeated use of neurotoxic insecticides against D. citri has resulted in the development of insecticide resistance. We evaluated the effects of the juvenile hormone analog methoprene on egg hatch, nymphal development, adult emergence, reproduction and behavior of D. citri. Methoprene significantly reduced the viability of eggs that were between 0 and 4 days old. Egg hatch of 0-48-h-old and 49-96-h-old eggs was 8 and 9%, respectively, when treated with 320 µg mL(-1) of methoprene. Methoprene caused significant mortality of first-, third- and fifth-instar D. citri nymphs and reduced adult emergence as compared with controls. Methoprene caused less than 5% adult emergence when first- and third-instar stages were treated, respectively, and less than 40% adult emergence when fifth instars were treated. Reduced fertility of females was observed when they emerged from methoprene-treated fifth instars. Methoprene was effective in reducing egg hatch, suppressing nymphal development and decreasing adult emergence of D. citri under laboratory conditions. Treatment of fifth instars reduced the fertility of females. Methoprene might be a possible tool for integrated management of D. citri. © 2015 Society of Chemical Industry.

Population dynamics and growth rates of endosymbionts during Diaphorina citri (Hemiptera, Liviidae) ontogeny.

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Dossi, Fabio Cleisto Alda; da Silva, Edney Pereira; Cônsoli, Fernando Luis

2014-11-01

The infection density of symbionts is among the major parameters to understand their biological effects in host-endosymbionts interactions. Diaphorina citri harbors two bacteriome-associated bacterial endosymbionts (Candidatus Carsonella ruddii and Candidatus Profftella armatura), besides the intracellular reproductive parasite Wolbachia. In this study, the density dynamics of the three endosymbionts associated with the psyllid D. citri was investigated by real-time quantitative PCR (qPCR) at different developmental stages. Bacterial density was estimated by assessing the copy number of the 16S rRNA gene for Carsonella and Profftella, and of the ftsZ gene for Wolbachia. Analysis revealed a continuous growth of the symbionts during host development. Symbiont growth and rate curves were estimated by the Gompertz equation, which indicated a negative correlation between the degree of symbiont-host specialization and the time to achieve the maximum growth rate (t*). Carsonella densities were significantly lower than those of Profftella at all host developmental stages analyzed, even though they both displayed a similar trend. The growth rates of Wolbachia were similar to those of Carsonella, but Wolbachia was not as abundant. Adult males displayed higher symbiont densities than females. However, females showed a much more pronounced increase in symbiont density as they aged if compared to males, regardless of the incorporation of symbionts into female oocytes and egg laying. The increased density of endosymbionts in aged adults differs from the usual decrease observed during host aging in other insect-symbiont systems.

Attractiveness of Host Plant Volatile Extracts to the Asian Citrus Psyllid, Diaphorina citri, is Reduced by Terpenoids from the Non-Host Cashew.

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Fancelli, Marilene; Borges, Miguel; Laumann, Raul A; Pickett, John A; Birkett, Michael A; Blassioli-Moraes, Maria C

2018-04-01

Diaphorina citri is a vector of the bacterial causative agent of Huanglongbing (HLB = Citrus greening), a severe disease affecting citrus crops. As there is no known control for HLB, manipulating insect behaviour through deployment of semiochemicals offers a promising opportunity for protecting citrus crops. The behavioural responses of D. citri to plant volatiles, and the identity of these plant volatiles were investigated. Volatiles were collected from host plants Murraya paniculata, Citrus sinensis, C. reshni, C. limettioides, Poncirus trifoliata, and from non-host plants Psidium guajava, Mangifera indica, Anacardium occidentale. In behavioural assays, female D. citri spent more time in the arms containing volatiles from either M. paniculata or C. sinensis compared to the control arms. When D. citri was exposed to volatiles collected from A. occidentale, they preferred the control arm. Volatiles emitted from the other studied plants did not influence the foraging behaviour of D. citri. Chemical analyses of volatile extracts from C. sinensis, M. paniculata, and A. occidentale revealed the presence of the terpenoids (E)-4,8-dimethylnona-1,3,7-triene (DMNT) and (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene (TMTT) in higher amounts in A. occidentale. In further behavioural bioassays, female D. citri spent less time in arms containing a synthetic blend of DMNT and TMTT compared to the control arms. Female D. citri also spent less time in arms containing the synthetic blend in combination with volatile extracts from either M. paniculata or C. sinensis compared to the control arms. Results suggest that higher release of the two terpenoids by A. occidentale make this species unattractive to D. citri, and that the terpenoids could be used in reducing colonisation of citrus plants and therefore HLB infection.

Double-stranded RNA delivery through soaking mediates silencing of the muscle protein 20 and increases mortality to the Asian citrus psyllid, Diaphorina citri.

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Yu, Xiudao; Gowda, Siddarame; Killiny, Nabil

2017-09-01

Asian citrus psyllid, Diaphorina citri Kuwayama, is the most important economic pest of citrus because it transmits Candidatus Liberibacter asiaticus (CLas), the causal agent of huanglongbing (HLB). Silencing genes by RNA interference (RNAi) is a promising approach for controlling D. citri. RNAi-based insect management strategies depend on the selection of suitable target genes. The muscle protein 20 gene DcMP20 was characterized from D. citri in an effort to impair proper muscle development through RNAi. Phylogenetic analysis showed that DcMP20 was more closely related to MP20 from Drosophila compared with its counterpart from other insect species. Developmental expression analysis revealed that transcription of DcMP20 was development dependent and reached a maximum level in the last instar (fourth-fifth) of the nymphal stage. The extent of RNAi in D. citri was dose dependent, with dsRNA-DcMP20 at 75 ng µL -1 being sufficient to knock down endogenous DcMP20 expression, which resulted in significant mortality and reduced body weight that positively correlated with the silencing of DcMP20. No effect was found when dsRNA-GFP or water was used, indicating the specific effect of dsRNA-DcMP20. Our results suggest that dsRNA can be delivered to D. citri through soaking, and DcMP20 is an effective RNAi target to be used in the management of D. citri. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

RNA interference of acetylcholinesterase in the Asian citrus psyllid, Diaphorina citri, increases its susceptibility to carbamate and organophosphate insecticides.

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Kishk, Abdelaziz; Hijaz, Faraj; Anber, Helmy A I; AbdEl-Raof, Tsamoh K; El-Sherbeni, AbdEl-Hakeem D; Hamed, Sobhy; Killiny, Nabil

2017-11-01

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Lividae) transmits the Candidatus Liberibacter asiaticus, which causes citrus greening disease or Huanglongbing, (HLB). To date, there is no efficient cure for HLB disease and the control of D. citri using insecticides became the most important tools for the management of HLB. However, the extensive use of insecticides could increase D. citri resistance to these insecticides. The objective of this study was to investigate the effect of RNA interference of acetylcholinesterase (AChE) on the mortality and susceptibility of D. citri to the four major insecticides used in Florida. In this study, we used a consensus sequence derived from the two AChE genes and cholinesterase 2-like (ChE-2-like) gene to target all of the three genes. Treatment with dsRNA-AChE increased the mortality percentages of both nymphs and adults of D. citri. The mortality percentage increased with the increase in the concentration of applied dsRNA-AChE, and the highest mortality (> 60%) was observed at the highest applied concentration (125ng/μl). Treatments of nymphs or adults with dsRNA-AChE down-regulated the expression of the three targeted genes of D. citri. Silencing of AChE and ChE in D. citri nymphs increased the susceptibility of emerged adults to chlorpyrifos and carbaryl, which act as AChE inhibitors. However, treatment with dsRNA-AChE did not increase the susceptibility of emerged adults to imidacloprid, which acts as an agonist of nicotinic acetylcholine receptors. In the same manner, treatment of adults with dsRNA-AChE increased their susceptibility to chlorpyrifos and carbaryl, but did not affect their susceptibility to imidacloprid. The ANOVA did not show any significant increase in susceptibility of D. citri adults to fenpropathrin after treatment with dsRNA-AChE, either as nymphs or as adults. However, simple linear regression showed that treatment with dsRNA-AChE increased D. citri susceptibility to fenpropathrin

Susceptibilities of Candidatus Liberibacter asiaticus-infected and noninfected Diaphorina citri to entomopathogenic fungi and their detoxification enzyme activities under different temperatures.

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Hussain, Mubasher; Akutse, Komivi Senyo; Lin, Yongwen; Chen, Shiman; Huang, Wei; Zhang, Jinguan; Idrees, Atif; Qiu, Dongliang; Wang, Liande

2018-03-25

Some entomopathogenic fungi species, Isaria fumosorosea, and Hirsutella citriformis were found to be efficient against the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). However, the susceptibility to these fungi increases when the psyllid infected with Candidatus Liberibacter asiaticus (Las), which is transmitted by D. citri and causes citrus greening disease. In this study, we examined the Las-infected and Las-uninfected D. citri susceptibility to entomopathogenic fungi at different temperature regimes (5-40°C). When D. citri adults exposed to cold temperature (5°C), they showed less susceptibility to entomopathogenic fungi as compared with control (27°C). Irrespective of infection with Las, a significantly positive correlation was observed between temperature and percentage mortality caused by different isolates of I. fumosorosea, 3A Ifr, 5F Ifr, PS Ifr, and H. citriformis isolates, HC3D and 2H. In contrast, a significantly negative correlation was found between temperature and percentage mortality for 3A Ifr for both Las-infected and Las-uninfected psyllids. Detoxification enzymes, Glutathione S-transferase levels in D. citri showed a negative correlation, whereas cytochrome P450 and general esterase levels were not correlated with changes in temperature. These findings revealed that detoxification enzymes and general esterase levels are not correlated with altered susceptibility to entomopathogenic fungi at the different temperature regimes. Conclusively, temperature fluctuations tested appear to be a significant factor impacting the management strategies of D. citri using entomopathogenic fungi. © 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

Host plant affects morphometric variation of Diaphorina citri (Hemiptera: Liviidae

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Thomson M. Paris

2016-11-01

Full Text Available The Asian citrus psyllid (ACP, Diaphorina citri Kuwayama, is one of the most serious citrus pests worldwide due to its role as vector of huanglongbing or citrus greening disease. While some optimal plant species for ACP oviposition and development have been identified, little is known of the influence of host plants on ACP size and shape. Our goal was to determine how size and shape of ACP wing and body size varies when development occurs on different host plants in a controlled rearing environment. ACP were reared on six different rutaceous species; Bergera koenigii, Citrus aurantifolia, Citrus macrophylla, Citrus maxima, Citrus taiwanica and Murraya paniculata. Adults were examined for morphometric variation using traditional and geometric analysis based on 12 traits or landmarks. ACP reared on C. taiwanica were consistently smaller than those reared on the other plant species. Wing aspect ratio also differed between C. maxima and C. taiwanica. Significant differences in shape were detected with those reared on M. paniculata having narrower wings than those reared on C. macrophylla. This study provides evidence of wing size and shape differences of ACP based on host plant species which potentially may impact dispersal. Further study is needed to determine if behavioral and physiological differences are associated with the observed phenotypic differences.

Host plant affects morphometric variation of Diaphorina citri (Hemiptera: Liviidae).

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Paris, Thomson M; Allan, Sandra A; Hall, David G; Hentz, Matthew G; Hetesy, Gabriella; Stansly, Philip A

2016-01-01

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is one of the most serious citrus pests worldwide due to its role as vector of huanglongbing or citrus greening disease. While some optimal plant species for ACP oviposition and development have been identified, little is known of the influence of host plants on ACP size and shape. Our goal was to determine how size and shape of ACP wing and body size varies when development occurs on different host plants in a controlled rearing environment. ACP were reared on six different rutaceous species; Bergera koenigii , Citrus aurantifolia , Citrus macrophylla , Citrus maxima , Citrus taiwanica and Murraya paniculata . Adults were examined for morphometric variation using traditional and geometric analysis based on 12 traits or landmarks. ACP reared on C. taiwanica were consistently smaller than those reared on the other plant species. Wing aspect ratio also differed between C. maxima and C. taiwanica . Significant differences in shape were detected with those reared on M. paniculata having narrower wings than those reared on C. macrophylla . This study provides evidence of wing size and shape differences of ACP based on host plant species which potentially may impact dispersal. Further study is needed to determine if behavioral and physiological differences are associated with the observed phenotypic differences.

Citrus tristeza virus-based RNAi in citrus plants induces gene silencing in Diaphorina citri, a phloem-sap sucking insect vector of citrus greening disease (Huanglongbing).

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Hajeri, Subhas; Killiny, Nabil; El-Mohtar, Choaa; Dawson, William O; Gowda, Siddarame

2014-04-20

A transient expression vector based on Citrus tristeza virus (CTV) is unusually stable. Because of its stability it is being considered for use in the field to control Huanglongbing (HLB), which is caused by Candidatus Liberibacter asiaticus (CLas) and vectored by Asian citrus psyllid, Diaphorina citri. In the absence of effective control strategies for CLas, emphasis has been on control of D. citri. Coincident cohabitation in phloem tissue by CLas, D. citri and CTV was exploited to develop a novel method to mitigate HLB through RNA interference (RNAi). Since CTV has three RNA silencing suppressors, it was not known if CTV-based vector could induce RNAi in citrus. Yet, expression of sequences targeting citrus phytoene desaturase gene by CTV-RNAi resulted in photo-bleaching phenotype. CTV-RNAi vector, engineered with truncated abnormal wing disc (Awd) gene of D. citri, induced altered Awd expression when silencing triggers ingested by feeding D. citri nymphs. Decreased Awd in nymphs resulted in malformed-wing phenotype in adults and increased adult mortality. This impaired ability of D. citri to fly would potentially limit the successful vectoring of CLas bacteria between citrus trees in the grove. CTV-RNAi vector would be relevant for fast-track screening of candidate sequences for RNAi-mediated pest control. Copyright © 2014. Published by Elsevier B.V.

Two-Spotted Ladybeetle Adalia bipunctata L. (Coleoptera: Coccinellidae): A Commercially Available Predator to Control Asian Citrus Psyllid Diaphorina citri (Hemiptera: Liviidae).

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Khan, Azhar A; Qureshi, Jawwad A; Afzal, Muhammad; Stansly, Philip A

2016-01-01

The Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Liviidae) is an economically important pest of citrus because it serves as a vector of the causal pathogens of huanglongbing (HLB) also known as citrus greening disease. The increased use of insecticides for control of D. citri negatively impacts several natural enemies including some effective ladybeetle species which are not available commercially. The two-spotted ladybeetle, Adalia bipunctata (Coleoptera: Coccinellidae) is found in some crop and forest ecosystems of Asia, Europe and North America and available commercially. It is known to attack aphids and mealybugs but there are no published records of feeding on psyllids. We evaluated suitability and preference of A. bipunctata for nymphs of D. citri compared to corn leaf aphid Rhopalosiphum maidis (Hemiptera: Aphididae) a global pest of cereal crops and prey for many predaceous insects. We also compared development and reproduction of A. bipunctata on these two species with frozen eggs of the Mediterranean flour moth Ephestia kuehniella (Lepidoptera: Pyralidae) at 25°C. Initially, more D. citri than R. maidis nymphs were consumed in the no-choice tests although final consumption by larva and adult of A. bipunctata did not differ in the choice and no-choice tests. Larval development was prolonged by one day on D. citri compared to R. maidis nymphs but did not differ between either of these diets and E. kuehniella. Larval survival to adult averaged 93-100% and was not impacted by diet. Adult life span did not differ between diets although those on D. citri and R. maidis nymphs weighed less and produced fewer but more fertile eggs than on E. kuehniella eggs. Significant reduction of D. citri nymphs averaging 54% was observed in colonies caged with adult A. bipunctata on field planted citrus. R° (net reproductive rate) was least for beetles fed R. maidis, but otherwise there were no significant differences in demographic parameters. Successful

Effect of UV-Blocking Plastic Films on Take-Off and Host Plant Finding Ability of Diaphorina citri (Hemiptera: Liviidae).

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Miranda, M P; Dos Santos, F L; Felippe, M R; Moreno, A; Fereres, A

2015-02-01

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is a major pest of citrus worldwide due to its ability to transmit the bacteria associated with huanglongbing. Vision, behavior, and performance of insect pests can be manipulated by using ultraviolet (UV)-blocking materials. Thus, the aim of our study was to evaluate how UV-blocking plastic films may affect the take-off and host plant finding ability of D. citri. To assess the effect of a UV-deficient environment on take-off, adult psyllids were released from a vial inside a screenhouse covered by a UV-blocking or standard (control) film and the number of insects remaining on each vial under each treatment was counted at different time intervals. Moreover, to assess the ability of D. citri to find citrus plants under a UV-deficient environment, two independent no-choice host plant finding assays with different plant arrangements were conducted. In each treatment, the number of psyllids per plant at different time intervals was counted. Both D. citri take-off and host plant finding ability was clearly disrupted under a UV-deficient environment. The number of psyllids remaining in the vials was significantly higher under UV-blocking than standard film in all periods recorded. Furthermore, psyllids were present in significantly higher number on citrus plants under standard film than under UV-blocking film in all of the periods assessed and experiments conducted. Our results showed that UV-blocking materials could become a valuable strategy for integrated management of D. citri and huanglongbing in citrus grown in enclosed environments. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

'Candidatus Liberibacter asiaticus' Accumulates inside Endoplasmic Reticulum Associated Vacuoles in the Gut Cells of Diaphorina citri.

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Ghanim, Murad; Achor, Diann; Ghosh, Saptarshi; Kontsedalov, Svetlana; Lebedev, Galina; Levy, Amit

2017-12-05

Citrus greening disease known also as Huanglongbing (HLB) caused by the phloem-limited bacterium 'Candidatus Liberibacter asiaticus' (CLas) has resulted in tremendous losses and the death of millions of trees worldwide. CLas is transmitted by the Asian citrus psyllid Diaphorina citri. The closely-related bacteria 'Candidatus Liberibacter solanacearum' (CLso), associated with vegetative disorders in carrots, is transmitted by the carrot psyllid Bactericera trigonica. A promising approach to prevent the transmission of these pathogens is to interfere with the vector-pathogen interactions, but our understanding of these processes is limited. It was recently reported that CLas induced changes in the nuclear architecture, and activated programmed cell death, in D. citri midgut cells. Here, we used electron and fluorescent microscopy and show that CLas induces the formation of endoplasmic reticulum (ER)-associated bodies. The bacterium recruits those ER structures into Liberibacter containing vacuoles (LCVs), in which bacterial cells seem to propagate. ER- associated LCV formation was unique to CLas, as we could not detect these bodies in B. trigonica infected with CLso. ER recruitment is hypothesized to generate a safe replicative body to escape cellular immune responses in the insect gut. Understanding the molecular interactions that undelay these responses will open new opportunities for controlling CLas.

Diaphorina citri Nymphs Are Resistant to Morphological Changes Induced by "Candidatus Liberibacter asiaticus" in Midgut Epithelial Cells.

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Mann, Marina; Fattah-Hosseini, Somayeh; Ammar, El-Desouky; Stange, Richard; Warrick, EricaRose; Sturgeon, Kasie; Shatters, Robert; Heck, Michelle

2018-04-01

" Candidatus Liberibacter asiaticus" is the causative bacterium associated with citrus greening disease. " Ca Liberibacter asiaticus" is transmitted by Diaphorina citri more efficiently when it is acquired by nymphs rather than adults. Why this occurs is not known. We compared midguts of D. citri insects reared on healthy or " Ca Liberibacter asiaticus"-infected citrus trees using quantitative PCR, confocal microscopy, and mitochondrial superoxide staining for evidence of oxidative stress. Consistent with its classification as propagative, " Ca Liberibacter asiaticus" titers were higher in adults than in nymphs. Our previous work showed that adult D. citri insects have basal levels of karyorrhexis (fragmentation of the nucleus) in midgut epithelial cells, which is increased in severity and frequency in response to " Ca Liberibacter asiaticus." Here, we show that nymphs exhibit lower levels of early-stage karyorrhexis than adults and are refractory to the induction of advanced karyorrhexis by " Ca Liberibacter asiaticus" in the midgut epithelium. MitoSox Red staining showed that guts of infected adults, particularly males, experienced oxidative stress in response to " Ca Liberibacter asiaticus." A positive correlation between the titers of " Ca Liberibacter asiaticus" and the Wolbachia endosymbiont was observed in adult and nymph midguts, suggesting an interplay between these bacteria during development. We hypothesize that the resistance of the nymph midgut to late-stage karyorrhexis through as yet unknown molecular mechanisms benefits " Ca Liberibacter asiaticus" for efficient invasion of midgut epithelial cells, which may be a factor explaining the developmental dependency of " Ca Liberibacter asiaticus" acquisition by the vector.

Crystal structure of diaphorin methanol monosolvate isolated from Diaphorina citri Kuwayama, the insect vector of citrus greening disease

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D. Marian Szebenyi

2018-04-01

Full Text Available The title compound C22H39NO9·CH3OH [systematic name: (S-N-((S-{(2S,4R,6R-6-[(S-2,3-dihydroxypropyl]-4-hydroxy-5,5-dimethyltetrahydro-2H-pyran-2-yl}(hydroxymethyl-2-hydroxy-2-[(2R,5R,6R-2-methoxy-5,6-dimethyl-4-methylenetetrahydro-2H-pyran-2-yl]acetamide methanol monosolvate], was isolated from the Asian citrus psyllid, Diaphorina citri Kuwayama, and crystallizes in the space group P21. `Candidatus Profftella armatura' a bacterial endosymbiont of D. citri, biosynthesizes diaphorin, which is a hybrid polyketide–nonribosomal peptide comprising two highly substituted tetrahydropyran rings joined by an N-acyl aminal bridge [Nakabachi et al. (2013. Curr. Biol. 23, 1478–1484]. The crystal structure of the title compound establishes the complete relative configuration of diaphorin, which agrees at all nine chiral centers with the structure of the methanol monosolvate of the di-p-bromobenzoate derivative of pederin, a biogenically related compound whose crystal structure was reported previously [Furusaki et al. (1968. Tetrahedron Lett. 9, 6301–6304]. Thus, the absolute configuration of diaphorin is proposed by analogy to that of pederin.

Risk assessment of various insecticides used for management of Asian citrus psyllid, Diaphorina citri in Florida citrus, against honey bee, Apis mellifera.

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Chen, Xue Dong; Gill, Torrence A; Pelz-Stelinski, Kirsten S; Stelinski, Lukasz L

2017-04-01

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is a major pest of citrus trees worldwide. A wide variety of insecticides are used to manage D. citri populations within citrus groves in Florida. However, in areas shared by citrus growers and beekeepers the use of insecticides may increase the risks of Apis mellifera  L. (Hymenoptera: Apidae) loss and contaminated honey. The objective of this research was to determine the environmental toxicity of insecticides, spanning five different modes of action used to control D. citri, to A. mellifera. The insecticides investigated were imidacloprid, fenpropathrin, dimethoate, spinetoram and diflubenzuron. In laboratory experiments, LD 50 values were determined and ranged from 0.10 to 0.53 ng/μl for imidacloprid, fenpropathrin, dimethoate and spinetoram. LD 50 values for diflubenzuron were >1000 ng/μl. Also, a hazard quotient was determined and ranged from 1130.43 to 10893.27 for imidacloprid, fenpropathrin, dimethoate, and spinetoram. This quotient was mellifera 3 and 7 days after application. Spinetoram and imidacloprid were moderately toxic to A. mellifera at the recommended rates for D. citri. Diflubenzuron was not toxic to A. mellifera in the field as compared with untreated control plots. Phenoloxidase (PO) activity of A. mellifera was higher than in untreated controls when A. mellifera were exposed to 14 days old residues. The results indicate that diflubenzuron may be safe to apply in citrus when A. mellifera are foraging, while most insecticides used for management of D. citri in citrus are likely hazardous under various exposure scenarios.

DIVERSIDAD GENÉTICA DE Diaphorina citri EN CULTIVOS CÍTRICOS DEL VALLE DEL CAUCA Y QUINDÍO (COLOMBIA

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MIGUEL ANGEL MONCAYO-DONOSO

Full Text Available El psílido asiático Diaphorina citri es el principal vector de la bacteria Candidatus Liberibacter que causa la enfermedad Huanglongbing, considerada devastadora en la citricultura mundial y aún no reportada en Colombia. La variabilidad genética poblacional de D. citri fue estudiadautilizando la técnica de secuenciación del gen COI mitocondrial como marcador molecular. Se realizaron colectas de adultos en zonas productoras de cítricos del Valle del Cauca y el Quindío. La amplificación se realizó con dos parejas de cebadores específicos para hemípteros. Los productos de la PCR fueron secuenciados en Macrogen-Korea obteniéndose un total de 124 secuencias. En el análisis bioinformático se utilizaron los programas Vector NTI 11.5, Arlequín V 3.5, MEGA 5 y MAFFT 6. Los índices de diversidad molecular fueron similares entre poblaciones, mostrando un mismo origen, separación reciente de poblaciones y que no existe diferenciación genética significativa que pueda albergar variantes de la bacteria; sin embargo, el índice de diversidad haplotípica fue más alto que el de diversidad nucleotídica y este último índice presentó bajo número de sitios polimórficos, indicando que las poblaciones de D. citri están en expansión. El estudio de la variabilidad genética del vector representa una herramienta para predecir escenarios probables de propagación de la enfermedad.

Behavioral Plasticity in Probing by Diaphorina citri (Hemiptera, Liviidae): Ingestion from Phloem Versus Xylem is Influenced by Leaf Age and Surface.

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Ebert, Timothy A; Backus, Elaine A; Shugart, Holly J; Rogers, Michael E

2018-01-01

Diaphorina citri is a major pest of citrus because it transmits Candidatus Liberibacter asiaticus, a phloem-limited bacterium that putatively causes Huanglongbing (HLB). The disease moves slowly through a tree, and the vector facilitates further within-tree movement via transmission of the pathogen. However, this only happens when D. citri stylets contact the phloem, to inoculate bacteria during phloem salivation and acquire bacteria during phloem sap ingestion. Behavioral changes in D. citri associated with different plant parts would affect how long it takes to reach phloem and how long the psyllids stays in phloem to ingest, thereby influencing the risk of disease spread. D. citri feeding was recorded on the abaxial and adaxial surfaces of mature and immature citrus leaves. Adults in the field can be found on these surfaces at all times of year. On abaxial surface of immature leaves, phloem salivation would occur after 11 h on average, but rarely as soon as 0.56 h. The corresponding values on mature leaves were 16 and 2.7. In general, psyllids spent more time ingesting phloem sap on immature leaves than on mature leaves. Psyllids on abaxial surfaces spent more time ingesting from phloem, though the strength of this effect was less than for immature versus mature leaves. In contrast, xylem ingestion increased on mature leaves compared with young. The biological differences that could produce this outcome are discussed. The results discussed herein are of relevance to further studies on the efficacy of an insecticide to act quickly enough to prevent pathogen transmission.

Crystal structure of diaphorin methanol monosolvate isolated from Diaphorina citri Kuwayama, the insect vector of citrus greening disease.

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Szebenyi, D Marian; Kriksunov, Irina; Howe, Kevin J; Ramsey, John S; Hall, David G; Heck, Michelle L; Krasnoff, Stuart B

2018-04-01

The title compound C 22 H 39 NO 9 ·CH 3 OH [systematic name: ( S )- N -(( S )-{(2 S ,4 R ,6 R )-6-[( S )-2,3-di-hydroxy-prop-yl]-4-hy-droxy-5,5-di-methyl-tetra-hydro-2 H -pyran-2-yl}(hy-droxy)meth-yl)-2-hy-droxy-2-[(2 R ,5 R ,6 R )-2-meth-oxy-5,6-dimeth-yl-4-methyl-ene-tetra-hydro-2 H -pyran-2-yl]acetamide methanol monosolvate], was isolated from the Asian citrus psyllid, Diaphorina citri Kuwayama, and crystallizes in the space group P 2 1 . ' Candidatus Profftella armatura' a bacterial endosymbiont of D. citri , biosynthesizes diaphorin, which is a hybrid polyketide-nonribosomal peptide comprising two highly substituted tetra-hydro-pyran rings joined by an N -acyl aminal bridge [Nakabachi et al. (2013 ▸). Curr. Biol. 23 , 1478-1484]. The crystal structure of the title compound establishes the complete relative configuration of diaphorin, which agrees at all nine chiral centers with the structure of the methanol monosolvate of the di- p -bromo-benzoate derivative of pederin, a biogenically related compound whose crystal structure was reported previously [Furusaki et al. (1968 ▸). Tetra-hedron Lett. 9 , 6301-6304]. Thus, the absolute configuration of diaphorin is proposed by analogy to that of pederin.

Odorants for surveillance and control of the Asian Citrus Psyllid (Diaphorina citri.

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Iliano V Coutinho-Abreu

Full Text Available The Asian Citrus Psyllid (ACP, Diaphorina citri, can transmit the bacterium Candidatus Liberibacter while feeding on citrus flush shoots. This bacterium causes Huanglongbing (HLB, a major disease of citrus cultivation worldwide necessitating the development of new tools for ACP surveillance and control. The olfactory system of ACP is sensitive to variety of odorants released by citrus plants and offers an opportunity to develop new attractants and repellents.In this study, we performed single-unit electrophysiology to identify odorants that are strong activators, inhibitors, and prolonged activators of ACP odorant receptor neurons (ORNs. We identified a suite of odorants that activated the ORNs with high specificity and sensitivity, which may be useful in eliciting behavior such as attraction. In separate experiments, we also identified odorants that evoked prolonged ORN responses and antagonistic odorants able to suppress neuronal responses to activators, both of which can be useful in lowering attraction to hosts. In field trials, we tested the electrophysiologically identified activating odorants and identified a 3-odor blend that enhances trap catches by ∼230%.These findings provide a set of odorants that can be used to develop affordable and safe odor-based surveillance and masking strategies for this dangerous pest insect.

Temporal progression of 'Candidatus Liberibacter asiaticus' infection in citrus and acquisition efficiency by Diaphorina citri.

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Coletta-Filho, Helvecio D; Daugherty, Matthew P; Ferreira, Cléderson; Lopes, João R S

2014-04-01

Over the last decade, the plant disease huanglongbing (HLB) has emerged as a primary threat to citrus production worldwide. HLB is associated with infection by phloem-limited bacteria ('Candidatus Liberibacter' spp.) that are transmitted by the Asian citrus psyllid, Diaphorina citri. Transmission efficiency varies with vector-related aspects (e.g., developmental stage and feeding periods) but there is no information on the effects of host-pathogen interactions. Here, acquisition efficiency of 'Candidatus Liberibacter asiaticus' by D. citri was evaluated in relation to temporal progression of infection and pathogen titer in citrus. We graft inoculated sweet orange trees with 'Ca. L. asiaticus'; then, at different times after inoculation, we inspected plants for HLB symptoms, measured bacterial infection levels (i.e., titer or concentration) in plants, and measured acquisition by psyllid adults that were confined on the trees. Plant infection levels increased rapidly over time, saturating at uniformly high levels (≈10(8) copy number of 16S ribosomal DNA/g of plant tissue) near 200 days after inoculation-the same time at which all infected trees first showed disease symptoms. Pathogen acquisition by vectors was positively associated with plant infection level and time since inoculation, with acquisition occurring as early as the first measurement, at 60 days after inoculation. These results suggest that there is ample potential for psyllids to acquire the pathogen from trees during the asymptomatic phase of infection. If so, this could limit the effectiveness of tree rouging as a disease management tool and would likely explain the rapid spread observed for this disease in the field.

Compatibility and Efficacy of Isaria fumosorosea with Horticultural Oils for Mitigation of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae

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Vivek Kumar

2017-10-01

Full Text Available Horticultural oils are an important component of integrated management programs of several phytophagous arthropods and pathogens affecting fruit, ornamentals and vegetables in greenhouse and field production systems. Although effective against the target pest, their incompatibility with biological control agents can compromise efforts to develop eco-friendly management programs for important agricultural pests. In this study, we assessed the in vitro effect of selected refined petroleum oils used in citrus and other horticultural crops with a biopesticide containing the entomopathogenic fungi, Isaria fumosorosea (PFR-97 under laboratory conditions. Further, we used leaf disk bioassays to evaluate the combined efficacy of petroleum oils and I. fumosorosea against the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae, a major pest of citrus in the United States. All five petroleum oil treatments (Orchex, Sun Pure, Conoco Blend -1, Conoco Blend -2, and JMS were compatible with I. fumosorosea blastospores, as none of them were found to affect I. fumosorosea colony-forming units and radial fungal growth measured at 3, 6, 9, and 12 days post-inoculation. All mixed treatments performed better than I. fumosorosea alone against D. citri, where the highest mean survival time of D. citri was 12.5 ± 0.7 days. No significant differences in D. citri survival time and I. fumosorosea growth (fungal development index on dead cadavers, which is important for determining their horizontal transmission, were observed when mixed with Orchex, Sun Pure, Conoco Blend -2, and JMS. Results indicated that horticultural oils in combination with I. fumosorosea could offer citrus growers an alternative treatment for integrating into their current management programs while battling against D. citri in citrus production systems. Due to their eco-friendly, broad-spectrum effect, it could provide control against various citrus pests, while also encouraging the

Compatibility and Efficacy of Isaria fumosorosea with Horticultural Oils for Mitigation of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae).

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Kumar, Vivek; Avery, Pasco B; Ahmed, Juthi; Cave, Ronald D; McKenzie, Cindy L; Osborne, Lance S

2017-10-31

Horticultural oils are an important component of integrated management programs of several phytophagous arthropods and pathogens affecting fruit, ornamentals and vegetables in greenhouse and field production systems. Although effective against the target pest, their incompatibility with biological control agents can compromise efforts to develop eco-friendly management programs for important agricultural pests. In this study, we assessed the in vitro effect of selected refined petroleum oils used in citrus and other horticultural crops with a biopesticide containing the entomopathogenic fungi, Isaria fumosorosea (PFR-97) under laboratory conditions. Further, we used leaf disk bioassays to evaluate the combined efficacy of petroleum oils and I. fumosorosea against the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae), a major pest of citrus in the United States. All five petroleum oil treatments (Orchex, Sun Pure, Conoco Blend -1, Conoco Blend -2, and JMS) were compatible with I. fumosorosea blastospores, as none of them were found to affect I. fumosorosea colony-forming units and radial fungal growth measured at 3, 6, 9, and 12 days post-inoculation. All mixed treatments performed better than I. fumosorosea alone against D. citri , where the highest mean survival time of D. citri was 12.5 ± 0.7 days. No significant differences in D. citri survival time and I. fumosorosea growth (fungal development index) on dead cadavers, which is important for determining their horizontal transmission, were observed when mixed with Orchex, Sun Pure, Conoco Blend -2, and JMS. Results indicated that horticultural oils in combination with I. fumosorosea could offer citrus growers an alternative treatment for integrating into their current management programs while battling against D. citri in citrus production systems. Due to their eco-friendly, broad-spectrum effect, it could provide control against various citrus pests, while also encouraging the retention of effective

Host range testing of Tamarixia radiata (Hymenoptera: Eulophidae) sourced from the Punjab of Pakistan for classical biological control of Diaphorina citri (Hemiptera: Liviidae: Euphyllurinae: Diaphorinini) in California.

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Hoddle, Mark S; Pandey, Raju

2014-02-01

ABSTRACT Tests evaluating the host range of Tamarixia radiata (Waterson) (Hymenoptera: Eulophidae), a parasitoid of the pestiferous Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), sourced from the Punjab of Pakistan, were conducted in quarantine at the University of California, Riverside, CA. Seven nontarget psyllid species (five native and two self-introduced species) representing five families were exposed to T radiata under the following three different exposure scenarios: 1) sequential no-choice tests, 2) static no-choice tests, and 3) choice tests. Nontarget species were selected for testing based on the following criteria: 1) taxonomic relatedness to the target, D. citri; 2) native psyllids inhabiting native host plants related to citrus that could release volatiles attractive to T. radiata; 3) native psyllids with a high probability of occurrence in native vegetation surrounding commercial citrus groves that could be encountered by T. radiata emigrating from D. citri-infested citrus orchards; 4) a common native pest psyllid species; and 5) a beneficial psyllid attacking a noxious weed. The results of host range testing were unambiguous; T radiata exhibited a narrow host range and high host specificity, with just one species of nontarget psyllid, the abundant native pest Bactericera cockerelli Sulc, being parasitized at low levels (citri poses negligible environmental risk.

Innate and Conditioned Responses to Chemosensory and Visual Cues in Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae), Vector of Huanglongbing Pathogens.

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Patt, Joseph M; Stockton, Dara; Meikle, William G; Sétamou, Mamoudou; Mafra-Neto, Agenor; Adamczyk, John J

2014-11-19

Asian citrus psyllid (Diaphorina citri) transmits Huanglongbing, a devastating disease that threatens citrus trees worldwide. A better understanding of the psyllid's host-plant selection process may lead to the development of more efficient means of monitoring it and predicting its movements. Since behavioral adaptations, such as associative learning, may facilitate recognition of suitable host-plants, we examined whether adult D. citri could be conditioned to visual and chemosensory stimuli from host and non-host-plant sources. Response was measured as the frequency of salivary sheaths, the residue of psyllid probing activity, in a line of emulsified wax on the surface of a test arena. The psyllids displayed both appetitive and aversive conditioning to two different chemosensory stimuli. They could also be conditioned to recognize a blue-colored probing substrate and their response to neutral visual cues was enhanced by chemosensory stimuli. Conditioned psyllids were sensitive to the proportion of chemosensory components present in binary mixtures. Naïve psyllids displayed strong to moderate innate biases to several of the test compounds. While innate responses are probably the psyllid's primary behavioral mechanism for selecting host-plants, conditioning may enhance its ability to select host-plants during seasonal transitions and dispersal.

Patogenicidade de Beauveria bassiana ao psilídeo Diaphorina citri e compatibilidade do fungo com produtos fitossanitários

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Ana Paula Ferreira Pinto

2012-12-01

Full Text Available O objetivo deste trabalho foi avaliar a patogenicidade de Beauveria bassiana a ninfas de Diaphorina citri (Hemiptera: Psyllidae e verificar a compatibilidade do fungo com produtos fitossanitários e sua persistência em plantas de citros. Ninfas de D. citri foram pulverizadas com B. bassiana, nas concentrações 5x10(6, 1x10(7, 5x10(7, 1x10(8, 5x10(8 e 1x10(9 conídios mL-1, para determinação da concentração letal. Para avaliação da compatibilidade do fungo com produtos fitossanitários, extrato de nim e cinco inseticidas de quatro grupos químicos diferentes foram incorporados individualmente ao meio de cultura BDA em que o fungo foi cultivado. Avaliaram-se o crescimento vegetativo, a esporulação e a viabilidade do entomopatógeno. Plantas de citros, mantidas em casa de vegetação, foram tratadas primeiramente com os produtos fitossanitários e depois com o entomopatógeno. Avaliaram-se os tempos de exposição de 24 horas e de 7 e 14 dias. O fungo foi patogênico às ninfas de D. citri; a CL50 foi de 0,4x10(7 e a CL90 de 6,7x10(7 conidios mL-1, no décimo dia de avaliação. Em laboratório, os produtos fitosssanitários reduzem o crescimento do fungo. Em casa de vegetação, os produtos não afetam a sobrevivência do fungo nas plantas de citros.

Acute Toxicity of Fresh and Aged Residues of Pesticides to the Parasitoid Tamarixia radiata and to the HLB-Bacteria Vector Diaphorina citri.

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Beloti, V H; Alves, G R; Moral, R A; Demétrio, C G B; Yamamoto, P T

2018-06-01

One method for controlling the Asian citrus psyllid (ACP) Diaphorina citri Kuwayama, the vector of the putative causal agent of Huanglongbing, uses the parasitoid Tamarixia radiata (Waterston). However, the general intensive use of insecticides has reduced the numbers of this parasitoid. This study evaluated the effect of the residual action of 24 insecticides on T. radiata and also determined the differential toxicity of insecticides to D. citri and T. radiata, using three bioassays. In the first, when adults of the parasitoid were exposed to residues of the 24 insecticides, ten were considered short-life (class 1), six slightly persistent (class 2), five moderately persistent (class 3), and three insecticides were considered persistent (class 4), under the IOBC/WPRS classification system. The second bioassay evaluated the sublethal concentrations of the persistent insecticides (formetanate, dimethoate, spinosad). Increasing the concentrations of the insecticides increased the number that were classified as persistent. In the third bioassay, evaluation of the differential toxicity of eight insecticides to the ACP and the parasitoid showed that chlorpyrifos and bifenthrin were more harmful to T. radiata. Therefore, these two insecticides are not recommended for application at the time of parasitoid release. Cypermethrin, imidacloprid, and dimethoate caused higher mortality of D. citri and are most often recommended in IPM programs. The choice of an insecticide for the control of citrus pests must be made with care, aiming to preserve the natural enemies in the ecosystem, and thereby contribute to the success of biological control.

Delivery of dsRNA through topical feeding for RNA interference in the citrus sap piercing-sucking hemipteran, Diaphorina citri.

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Killiny, Nabil; Kishk, Abdelaziz

2017-06-01

RNA interference (RNAi) is a powerful means to study functional genomics in insects. The delivery of dsRNA is a challenging step in the development of RNAi assay. Here, we describe a new delivery method to increase the effectiveness of RNAi in the Asian citrus psyllid Diaphorina citri. Bromophenol blue droplets were topically applied to fifth instar nymphs and adults on the ventral side of the thorax between the three pairs of legs. In addition to video recordings that showed sucking of the bromophenol blue by the stylets, dissected guts turned blue indicating that the uptake was through feeding. Thus, we called the method topical feeding. We targeted the abnormal wing disc gene (awd), also called nucleoside diphosphate kinase (NDPK), as a reporter gene to prove the uptake of dsRNA via this method of delivery. Our results showed that dsRNA-awd caused reduction of awd expression and nymph mortality. Survival and lifespan of adults emerged from treated nymphs and treated adults were affected. Silencing awd caused wing malformation in the adults emerged from treated nymphs. Topical feeding as a delivery of dsRNA is highly efficient for both nymphs and adults. The described method could be used to increase the efficiency of RNAi in D. citri and other sap piercing-sucking hemipterans. © 2017 Wiley Periodicals, Inc.

Innate and Conditioned Responses to Chemosensory and Visual Cues in Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae, Vector of Huanglongbing Pathogens

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Joseph M. Patt

2014-11-01

Full Text Available Asian citrus psyllid (Diaphorina citri transmits Huanglongbing, a devastating disease that threatens citrus trees worldwide. A better understanding of the psyllid’s host-plant selection process may lead to the development of more efficient means of monitoring it and predicting its movements. Since behavioral adaptations, such as associative learning, may facilitate recognition of suitable host-plants, we examined whether adult D. citri could be conditioned to visual and chemosensory stimuli from host and non-host-plant sources. Response was measured as the frequency of salivary sheaths, the residue of psyllid probing activity, in a line of emulsified wax on the surface of a test arena. The psyllids displayed both appetitive and aversive conditioning to two different chemosensory stimuli. They could also be conditioned to recognize a blue-colored probing substrate and their response to neutral visual cues was enhanced by chemosensory stimuli. Conditioned psyllids were sensitive to the proportion of chemosensory components present in binary mixtures. Naïve psyllids displayed strong to moderate innate biases to several of the test compounds. While innate responses are probably the psyllid’s primary behavioral mechanism for selecting host-plants, conditioning may enhance its ability to select host-plants during seasonal transitions and dispersal.

Morphological abnormalities and cell death in the Asian citrus psyllid (Diaphorina citri) midgut associated with Candidatus Liberibacter asiaticus.

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Ghanim, Murad; Fattah-Hosseini, Somayeh; Levy, Amit; Cilia, Michelle

2016-09-15

Candidatus Liberibacter asiaticus (CLas) is a phloem-limited, gram-negative, fastidious bacterium that is associated with the development of citrus greening disease, also known as Huanglongbing (HLB). CLas is transmitted by the Asian citrus psyllid (ACP) Diaphorina citri, in a circulative manner. Two major barriers to transmission within the insect are the midgut and the salivary glands. We performed a thorough microscopic analysis within the insect midgut following exposure to CLas-infected citrus trees. We observed changes in nuclear architecture, including pyknosis and karyorrhexis as well as changes to the actin cytoskeleton in CLas-exposed midgut cells. Further analyses showed that the changes are likely due to the activation of programmed cell death as assessed by Annexin V staining and DNA fragmentation assays. These results suggest that exposure to CLas-infected trees induces apoptotic responses in the psyllid midgut that should be further investigated. Understanding the adaptive significance of the apoptotic response has the potential to create new approaches for controlling HLB.

Resistance of Citrus and Related Genera to Diaphorina citri Kuwayama (Hemiptera: Liviidae).

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Borgoni, P C; Vendramim, J D; Lourencão, A L; Machado, M A

2014-10-01

The present study was developed to evaluate the resistance of the following genotypes of Citrus and related genera to this pest: 'Pera,' 'Natal', and 'Washington Navel' oranges (Citrus sinensis), 'Marsh Seedless' grapefruit (Citrus paradisi), hardy orange 'Rubidoux' (Poncirus trifoliata), kumquat (Fortunella margarita Swingle), citrumelo 'Swingle' (C. paradisi x P. trifoliata), and citrange 'Troyer' (P. trifoliata x C. sinensis). The experiments were performed in greenhouses with plants grafted onto 'Rangpur' lime (Citrus limonia) and placed individually in voile cages. The preference for oviposition in a no-choice test, and the effect of genotype were evaluated. The egg-adult cycle was monitored to determine the effect of genotype on the biology of the insect. Poncirus 'Rubidoux' was the least preferred genotype for oviposition; reduced number of eggs was also found to occur on citrange 'Troyer', and 'Marsh Seedless' was the genotype with the most eggs. No significant variation in the duration of the embryonic period was observed; however, a difference in the viability of eggs was found, with the lowest egg viabilities on 'Swingle.' Kumquat and 'Marsh Seedless' genotypes were correlated with increased durations of the nymphal phase, however, there was no difference in the survival of this phase. Fecundity of females on 'Troyer', 'Swingle', and kumquat was reduced. Considering all of the evaluated parameters, it was concluded that cultivars of sweet orange are the most susceptible genotypes to Diaphorina citri. Regarding oviposition, P. trifoliata 'Rubidoux' showed resistance of the antixenosis type.

Exposure to Guava Affects Citrus Olfactory Cues and Attractiveness to Diaphorina citri (Hemiptera: Psyllidae).

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Barman, Jagadish Chandra; Campbell, Stuart A; Zeng, Xinnian

2016-06-01

Intercropping can reduce agricultural pest incidence, and represents an important sustainable alternative to conventional pest control methods. Understanding the ecological mechanisms for intercropping could help optimize its use, particularly in tropical systems which present a large number of intercropping possibilities. Citrus is threatened worldwide by greening disease (huanglongbing, HLB) vectored by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae). Control of HLB and citrus psyllid can be partially achieved through intercropping with guava, Psidium guajava L., but the mechanisms remain unclear. We tested the hypothesis that guava olfactory cues affect psyllid behavior by altering the attractiveness of citrus through plant-plant interactions. In choice and no-choice cage experiments, psyllid settlement was reduced on citrus shoots that had been exposed to guava shoot odors for at least 2 h. In Y-tube olfactometer experiments, psyllids oriented to odors of unexposed, compared with guava-exposed, citrus shoots. These behavioral results indicate that a mechanism for the success of guava intercropping for sustainable, ecological disease management may be the indirect effect of guava on citrus attractiveness. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Cloning and expressing a highly functional and substrate specific farnesoic acid o-methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama).

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Van Ekert, Evelien; Shatters, Robert G; Rougé, Pierre; Powell, Charles A; Smagghe, Guy; Borovsky, Dov

2015-01-01

The Asian citrus psyllid, Diaphorina citri, transmits a phloem-limited bacterium, Candidatus 'Liberibacter' asiaticus that causes citrus greening disease. Because juvenile hormone (JH) plays an important role in adult and nymphal development, we studied the final steps in JH biosynthesis in D. citri. A putative JH acid methyltransferase ortholog gene (jmtD) and its cognate cDNA were identified by searching D. citri genome database. Expression analysis shows expression in all life stages. In adults, it is expressed in the head-thorax, (containing the corpora allata), and the abdomen (containing ovaries and male accessory glands). A 3D protein model identified the catalytic groove with catalytically active amino acids and the S-adenosyl methionine (SAM)-binding loop. The cDNA was expressed in Escherichia coli cells and the purified enzyme showed high preference for farnesoic acid (FA) and homoFA (kcat of 0.752 × 10(-3) and 0.217 × 10(-3) s(-1), respectively) as compared to JH acid I (JHA I) (cis/trans/cis; 2Z, 6E, 10cis), JHA III (2E, 6E, 10cis), and JHA I (trans/cis/cis; 2E, 2Z, 10cis) (kcat of 0.081 × 10(-3), 0.013 × 10(-3), and 0.003 × 10(-3) s(-1), respectively). This suggests that this ortholog is a DcFA-o-methyl transferase gene (fmtD), not a jmtD, and that JH biosynthesis in D. citri proceeds from FA to JH III through methyl farnesoate (MF). DcFA-o-MT does not require Ca(2+), Mg(2+) or Zn(2+), however, Zn(2+) (1 mM) completely inhibits the enzyme probably by binding H115 at the active groove. This represents the first purified FA-o-MT from Hemiptera with preferred biological activity for FA and not JHA.

Global mtDNA genetic structure and hypothesized invasion history of a major pest of citrus, Diaphorina citri (Hemiptera: Liviidae).

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Luo, Yufa; Agnarsson, Ingi

2018-01-01

The Asian citrus psyllid Diaphorina citri Kuwayama is a key pest of citrus as the vector of the bacterium causing the "huanglongbing" disease (HLB). To assess the global mtDNA population genetic structure, and possible dispersal history of the pest, we investigated genetic variation at the COI gene collating newly collected samples with all previously published data. Our dataset consists of 356 colonies from 106 geographic sites worldwide. High haplotype diversity (H-mean = 0.702 ± 0.017), low nucleotide diversity (π-mean = 0.003), and significant positive selection (Ka/Ks = 32.92) were observed. Forty-four haplotypes (Hap) were identified, clustered into two matrilines: Both occur in southeastern and southern Asia, North and South America, and Africa; lineages A and B also occur in eastern and western Asia, respectively. The most abundant haplotypes were Hap4 in lineage A (35.67%), and Hap9 in lineage B (41.29%). The haplotype network identified them as the ancestral haplotypes within their respective lineages. Analysis of molecular variance showed significant genetic structure ( F ST  = 0.62, p  analysis suggests geographic structuring. We hypothesize a southern and/or southeastern Asia origin, three dispersal routes, and parallel expansions of two lineages. The hypothesized first route involved the expansion of lineage B from southern Asia into North America via West Asia. The second, the expansion of some lineage A individuals from Southeast Asia into East Asia, and the third involved both lineages from Southeast Asia spreading westward into Africa and subsequently into South America. To test these hypotheses and gain a deeper understanding of the global history of D. citri , more data-rich approaches will be necessary from the ample toolkit of next-generation sequencing (NGS). However, this study may serve to guide such sampling and in the development of biological control programs against the global pest D. citri .

RNA interference of two glutathione S-transferase genes, Diaphorina citri DcGSTe2 and DcGSTd1, increases the susceptibility of Asian citrus psyllid (Hemiptera: Liviidae) to the pesticides fenpropathrin and thiamethoxam.

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Yu, Xiudao; Killiny, Nabil

2018-03-01

The Asian citrus psyllid, Diaphorina citri Kuwayama, is an important agricultural pest of citrus globally. Foliar application of chemical insecticides is the most widely used option for reducing D. citri populations. Knockdown of glutathione S-transferase (GST) in several insect species leads to increased susceptibility to insecticides; however, information about the detoxifying role of GST genes in D. citri is unavailable. Via a sequence homology search, we isolated and characterized three DcGST genes (DcGSTd1, DcGSTe1 and DcGSTe2) from D. citri. Phylogenetic analysis grouped DcGSTd1 into the delta class of GST genes, whereas DcGSTe1 and DcGSTe2 were clustered in the epsilon clade. Gene expression analysis revealed that chlorpyrifos treatment increased the mRNA levels of DcGSTe1 and fenpropathrin enhanced the expression level of DcGSTd1, while DcGSTe2 was significantly up-regulated after exposure to thiamethoxam at a dose of 30% lethal concentration (LC30). RNA interference (RNAi) of DcGSTe2 and DcGSTd1 followed by an insecticide bioassay increased the mortalities of thiamethoxam-treated psyllids by 23.0% and fenpropathrin-treated psyllids by 15.0%. In contrast, knockdown of DcGSTe1 did not significantly increase the susceptibility of D. citri to any of these three insecticides. Further, feeding with double-stranded RNA (dsDcGSTe2-d1) interfusion co-silenced DcGSTe2 and DcGSTd1 expression in D. citri, and led to an increase of susceptibility to both fenpropathrin and thiamethoxam. The findings suggest that DcGSTe2 and DcGSTd1 play unique roles in detoxification of the pesticides thiamethoxam and fenpropathrin. In addition, co-silencing by creating a well-designed dsRNA interfusion against multiple genes was a good RNAi strategy in D. citri. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Lethal and Sublethal Impacts of Acaricides on Tamarixia radiata (Hemiptera: Eulophidae), an Important Ectoparasitoid of Diaphorina citri (Hemiptera: Liviidae).

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Lira, A C S; Zanardi, O Z; Beloti, V H; Bordini, G P; Yamamoto, P T; Parra, J R P; Carvalho, G A

2015-10-01

The use of synthetic acaricides for management of pest mites may alter the efficacy of the ectoparasitoid Tamarixia radiata (Waterston) in biological control of Diaphorina citri Kuwayama, the vector of the bacteria associated with huanglongbing (HLB) in citrus orchards. We evaluated the toxicity of 16 acaricides that are recommended for the control of citrus-pest mites to T. radiata. Acrinathrin, bifenthrin, carbosulfan, and fenpropathrin caused high acute toxicity and were considered harmful (mortality >77%) to T. radiata. Abamectin, diflubenzuron, etoxazole, fenbutatin oxide, fenpyroximate, flufenoxuron, hexythiazox, propargite, spirodiclofen, and sulfur caused low acute toxicity and affected the parasitism rate and emergence rate of adults (F1 generation), and were considered slightly harmful to T. radiata. Dicofol and pyridaben did not affect the survival and action of the ectoparasitoid, and were considered harmless. In addition to its acute toxicity, carbosulfan caused mortality higher than 25% for >30 d after application, and was considered persistent. Acrinathrin, bifenthrin, fenpropathrin, fenpyroximate, propargite, and sulfur caused mortalities over 25% until 24 d after application and were considered moderately persistent; abamectin was slightly persistent, and fenbutatin oxide was short lived. Our results suggest that most acaricides used to control pest mites in citrus affect the density and efficacy of T. radiata in the biological control of D. citri. However, further evaluations are needed in order to determine the effect of these products on this ectoparasitoid under field conditions. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Dinâmica populacional e parasitismo natural de Diaphorina citri Kuwayama (Hemiptera: Liviidae em pomares de citros em Sergipe

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Adriano Farias

2018-04-01

Abstract. The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Sternorrhyncha: Liviidae, which is the vector of the Hunglongbing (HLB bacterium, has become a key citrus pest in Brazil. In addition to citrus, the ornamental plant known as Orange Jasmine, Murraya paniculata (L. Jack, also hosts the psyllid. The present work aimed at studying the population dynamics of the psyllid in citrus orchards and its parasitism in citrus and M. paniculata in the state of Sergipe, the fourth largest citrus-producing state in Brazil. The evaluations were performed fortnightly for eleven months in seven Pera Citrus sinensis (L. Osbeck orange orchards located in two municipalities of Sergipe state (Boquim, and Umbauba. The populations of eggs, nymphs and adults of D. citri were compared among all orchards and all developmental stages were related to the abiotic factors temperature, relative humidity and precipitation, in each locality. The psyllid was classified as an accessory species (not very abundant in the orchards of Sergipe, showing a higher population density in November, December and March. Precipitation was the only abiotic factor that contributed to the population increase of adults of the psyllid. High rates of parasitism (55% of psyllid nymphs by the exotic parasitoid Tamarixia radiata Waterston (Hymenoptera: Eulophidae were detected in M. paniculata plants. Based on the results, if the HLB bacterium is detected in Sergipe, shorter samplings should be performed in the spring and summer months aiming at vector management. In addition, T. radiata could be released inundatively in integrated vector management programs.

Antennal and Abdominal Transcriptomes Reveal Chemosensory Genes in the Asian Citrus Psyllid, Diaphorina citri.

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Wu, Zhongzhen; Zhang, He; Bin, Shuying; Chen, Lei; Han, Qunxin; Lin, Jintian

2016-01-01

The Asian citrus psyllid, Diaphorina citri is the principal vector of the highly destructive citrus disease called Huanglongbing (HLB) or citrus greening, which is a major threat to citrus cultivation worldwide. More effective pest control strategies against this pest entail the identification of potential chemosensory proteins that could be used in the development of attractants or repellents. However, the molecular basis of olfaction in the Asian citrus psyllid is not completely understood. Therefore, we performed this study to analyze the antennal and abdominal transcriptome of the Asian citrus psyllid. We identified a large number of transcripts belonging to nine chemoreception-related gene families and compared their expression in male and female adult antennae and terminal abdomen. In total, 9 odorant binding proteins (OBPs), 12 chemosensory proteins (CSPs), 46 odorant receptors (ORs), 20 gustatory receptors (GRs), 35 ionotropic receptors (IRs), 4 sensory neuron membrane proteins (SNMPs) and 4 different gene families encoding odorant-degrading enzymes (ODEs): 80 cytochrome P450s (CYPs), 12 esterase (ESTs), and 5 aldehyde dehydrogenases (ADE) were annotated in the D. citri antennal and abdominal transcriptomes. Our results revealed that a large proportion of chemosensory genes exhibited no distinct differences in their expression patterns in the antennae and terminal abdominal tissues. Notably, RNA sequencing (RNA-seq) data and quantitative real time-PCR (qPCR) analyses showed that 4 DictOBPs, 4 DictCSPs, 4 DictIRs, 1 DictSNMP, and 2 DictCYPs were upregulated in the antennae relative to that in terminal abdominal tissues. Furthermore, 2 DictOBPs (DictOBP8 and DictOBP9), 2 DictCSPs (DictOBP8 and DictOBP12), 4 DictIRs (DictIR3, DictIR6, DictIR10, and DictIR35), and 1 DictCYP (DictCYP57) were expressed at higher levels in the male antennae than in the female antennae. Our study provides the first insights into the molecular basis of chemoreception in this insect

Antennal and Abdominal Transcriptomes Reveal Chemosensory Genes in the Asian Citrus Psyllid, Diaphorina citri.

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Zhongzhen Wu

Full Text Available The Asian citrus psyllid, Diaphorina citri is the principal vector of the highly destructive citrus disease called Huanglongbing (HLB or citrus greening, which is a major threat to citrus cultivation worldwide. More effective pest control strategies against this pest entail the identification of potential chemosensory proteins that could be used in the development of attractants or repellents. However, the molecular basis of olfaction in the Asian citrus psyllid is not completely understood. Therefore, we performed this study to analyze the antennal and abdominal transcriptome of the Asian citrus psyllid. We identified a large number of transcripts belonging to nine chemoreception-related gene families and compared their expression in male and female adult antennae and terminal abdomen. In total, 9 odorant binding proteins (OBPs, 12 chemosensory proteins (CSPs, 46 odorant receptors (ORs, 20 gustatory receptors (GRs, 35 ionotropic receptors (IRs, 4 sensory neuron membrane proteins (SNMPs and 4 different gene families encoding odorant-degrading enzymes (ODEs: 80 cytochrome P450s (CYPs, 12 esterase (ESTs, and 5 aldehyde dehydrogenases (ADE were annotated in the D. citri antennal and abdominal transcriptomes. Our results revealed that a large proportion of chemosensory genes exhibited no distinct differences in their expression patterns in the antennae and terminal abdominal tissues. Notably, RNA sequencing (RNA-seq data and quantitative real time-PCR (qPCR analyses showed that 4 DictOBPs, 4 DictCSPs, 4 DictIRs, 1 DictSNMP, and 2 DictCYPs were upregulated in the antennae relative to that in terminal abdominal tissues. Furthermore, 2 DictOBPs (DictOBP8 and DictOBP9, 2 DictCSPs (DictOBP8 and DictOBP12, 4 DictIRs (DictIR3, DictIR6, DictIR10, and DictIR35, and 1 DictCYP (DictCYP57 were expressed at higher levels in the male antennae than in the female antennae. Our study provides the first insights into the molecular basis of chemoreception in this

Overview of worldwide diversity of Diaphorina citri Kuwayama mitochondrial cytochrome oxidase 1 haplotypes: two Old World lineages and a New World invasion

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Boykin, L.M.; De Barro, P.; Hall, D.G.; Hunter, W.B.; McKenzie, C.L.; Powell, C.A.; Shatters, R.G.

2012-01-01

Relationships among worldwide collections of Diaphorina citri (Asian citrus psyllid) were analyzed using mitochondrial cytochrome oxidase I (mtCOI) haplotypes from novel primers. Sequences were produced from PCR amplicons of an 821bp portion of the mtCOI gene using D. citri specific primers, derived from an existing EST library. An alignment was constructed using 612bps of this fragment and consisted of 212 individuals from 52 collections representing 15 countries. There were a total of eight polymorphic sites that separated the sequences into eight different haplotypes (Dcit-1 through Dcit-8). Phylogenetic network analysis using the statistical parsimony software, TCS, suggests two major haplotype groups with preliminary geographic bias between southwestern Asia (SWA) and southeastern Asia (SEA). The recent (within the last 15 to 25 years) invasion into the New World originated from only the SWA group in the northern hemisphere (USA and Mexico) and from both the SEA and SWA groups in the southern hemisphere (Brazil). In only one case, Reunion Island, did haplotypes from both the SEA and SWA group appear in the same location. In Brazil, both groups were present, but in separate locations. The Dcit-1 SWA haplotype was the most frequently encountered, including ~50% of the countries sampled and 87% of the total sequences obtained from India, Pakistan and Saudi Arabia. The second most frequently encountered haplotype, Dcit-2, the basis of the SEA group, represented ~50% of the countries and contained most of the sequences from Southeast Asia and China. Interestingly, only the Caribbean collections (Puerto Rico and Guadeloupe) represented a unique haplotype not found in other countries, indicating no relationship between the USA (Florida) and Caribbean introductions. There is no evidence for cryptic speciation for D. citri based on the COI region included in this study. PMID:22717059

Population diversity of Diaphorina citri (Hemiptera: Liviidae) in China based on whole mitochondrial genome sequences.

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Wu, Fengnian; Jiang, Hongyan; Beattie, G Andrew C; Holford, Paul; Chen, Jianchi; Wallis, Christopher M; Zheng, Zheng; Deng, Xiaoling; Cen, Yijing

2018-04-24

Diaphorina citri (Asian citrus psyllid; ACP) transmits 'Candidatus Liberibacter asiaticus' associated with citrus Huanglongbing (HLB). ACP has been reported in 11 provinces/regions in China, yet its population diversity remains unclear. In this study, we evaluated ACP population diversity in China using representative whole mitochondrial genome (mitogenome) sequences. Additional mitogenome sequences outside China were also acquired and evaluated. The sizes of the 27 ACP mitogenome sequences ranged from 14 986 to 15 030 bp. Along with three previously published mitogenome sequences, the 30 sequences formed three major mitochondrial groups (MGs): MG1, present in southwestern China and occurring at elevations above 1000 m; MG2, present in southeastern China and Southeast Asia (Cambodia, Indonesia, Malaysia, and Vietnam) and occurring at elevations below 180 m; and MG3, present in the USA and Pakistan. Single nucleotide polymorphisms in five genes (cox2, atp8, nad3, nad1 and rrnL) contributed mostly in the ACP diversity. Among these genes, rrnL had the most variation. Mitogenome sequences analyses revealed two major phylogenetic groups of ACP present in China as well as a possible unique group present currently in Pakistan and the USA. The information could have significant implications for current ACP control and HLB management. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

Male Diaphorina citri searching responses to vibrational communication signals

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Prototype devices have been developed that mimic D. citri female replies to male communication signals and lure males to a trap. The objective is to trap a high proportion of males that have landed on a host tree and have begun searching for females. This presentation describes the construction and ...

Species clarification of Isaria isolates used as biocontrol agents against Diaphorina citri (Hemiptera: Liviidae) in Mexico.

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Gallou, Adrien; Serna-Domínguez, María G; Berlanga-Padilla, Angélica M; Ayala-Zermeño, Miguel A; Mellín-Rosas, Marco A; Montesinos-Matías, Roberto; Arredondo-Bernal, Hugo C

2016-03-01

Entomopathogenic fungi belonging to the genus Isaria (Hypocreales: Cordycipitaceae) are promising candidates for microbial control of insect pests. Currently, the Mexican government is developing a biological control program based on extensive application of Isaria isolates against Diaphorina citri (Hemiptera: Liviidae), a vector of citrus huanglongbing disease. Previous research identified three promising Isaria isolates (CHE-CNRCB 303, 305, and 307; tentatively identified as Isaria fumosorosea) from Mexico. The goal of this work was to obtain a complete morphological and molecular characterization of these isolates. Comparative analysis of morphology established that the isolates showed similar characteristics to Isaria javanica. Multi-gene analysis confirmed the morphological identification by including the three isolates within the I. javanica clade. Additionally, this work demonstrated the misidentifications of three other Isaria isolates (CHE-CNRCB 310 and 324: I. javanica, formerly I. fumosorosea; CHE-CNRCB 393: I. fumosorosea, formerly Isaria farinosa), underlying the need for a full and correct characterization of an isolate before developing a biological control program. Finally, the inter-simple sequence repeat (ISSR) genotyping method revealed that the CHE-CNRCB 303, 305, and 307 isolates belong to three different genotypes. This result indicates that ISSR markers could be used as a tool to monitor their presence in field conditions. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Infection Density Dynamics of the Citrus Greening Bacterium “Candidatus Liberibacter asiaticus” in Field Populations of the Psyllid Diaphorina citri and Its Relevance to the Efficiency of Pathogen Transmission to Citrus Plants

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Ukuda-Hosokawa, Rie; Sadoyama, Yasutsune; Kishaba, Misaki; Kuriwada, Takashi; Anbutsu, Hisashi

2015-01-01

Huanglongbing, or citrus greening, is a devastating disease of citrus plants recently spreading worldwide, which is caused by an uncultivable bacterial pathogen, “Candidatus Liberibacter asiaticus,” and vectored by a phloem-sucking insect, Diaphorina citri. We investigated the infection density dynamics of “Ca. Liberibacter asiaticus” in field populations of D. citri with experiments using field-collected insects to address how “Ca. Liberibacter asiaticus” infection density in the vector insect is relevant to pathogen transmission to citrus plants. Of 500 insects continuously collected from “Ca. Liberibacter asiaticus”-infected citrus trees with pathological symptoms in the spring and autumn of 2009, 497 (99.4%) were “Ca. Liberibacter asiaticus” positive. The infections were systemic across head-thorax and abdomen, ranging from 103 to 107 bacteria per insect. In spring, the infection densities were low in March, at ∼103 bacteria per insect, increasing up to 106 to 107 bacteria per insect in April and May, and decreasing to 105 to 106 bacteria per insect in late May, whereas the infection densities were constantly ∼106 to 107 bacteria per insect in autumn. Statistical analysis suggested that several factors, such as insect sex, host trees, and collection dates, may be correlated with “Ca. Liberibacter asiaticus” infection densities in field D. citri populations. Inoculation experiments with citrus seedlings using field-collected “Ca. Liberibacter asiaticus”-infected insects suggested that (i) “Ca. Liberibacter asiaticus”-transmitting insects tend to exhibit higher infection densities than do nontransmitting insects, (ii) a threshold level (∼106 bacteria per insect) of “Ca. Liberibacter asiaticus” density in D. citri is required for successful transmission to citrus plants, and (iii) D. citri attaining the threshold infection level transmits “Ca. Liberibacter asiaticus” to citrus plants in a stochastic manner. These

Effects of Candidatus Liberibacter asiaticus on the fitness of the vector Diaphorina citri.

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Ren, S-L; Li, Y-H; Zhou, Y-T; Xu, W-M; Cuthbertson, A G S; Guo, Y-J; Qiu, B-L

2016-12-01

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama transmits the bacterium 'Candidatus Liberibacter asiaticus' (Las), which causes citrus huanglongbing (HLB) disease. Although many studies have been conducted on the biology of ACP on different host plants, few have taken the plant, Las bacteria and the vector insect within one context to evaluate the effects of Las on the fitness of ACP under field conditions. Understanding the relationship between Las and ACP is critical for both ACP and HLB disease management. We estimated the development and survival of ACP immatures, the longevity and fecundity of ACP female adults in four treatments (Las-positive or -negative ACP on Las-infected and -free citrus plants). Las-positive ACP immatures developed significantly faster on Las-infected citrus than those on Las-free plants. The fecundity and longevity of Las-positive female adults were also greater, or longer on Las-infected citrus shoots, whereas the survival of Las-positive immatures was significantly lower on Las-infected citrus shoots, compared to those that developed on Las-free plants. Similarly, the intrinsic rate of population increase (r m ) was highest (0·1404) when Las-positive ACP fed on Las-infected citrus shoots and the lowest (0·1328) when the Las-negative ACP fed on Las-free citrus shoots. Both the Las infection in ACP and citrus plants had obvious effects on the biology of ACP. When compared to the Las infection in ACP insects, the Las infection in citrus shoots had a more significant effect on the fitness of ACP. To efficiently prevent the occurrence and spread of HLB disease, it is critical to understand the ecological basis of vector outbreaks and disease incidence, especially under field conditions. Thus, this study has increased our understanding of the epidemiology of HLB transmitted by psyllids in nature. © 2016 The Society for Applied Microbiology.

Pengendalian Diaphorina citri (Vektor Penyakit CVPD dengan Metarrhizium anisopliae

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Kardi Raharjo

2000-07-01

and after insect infestation. The first phase of the research phase has been conducted in Temanggung, Completely Randomize Design (CRD factorial with three time replication. Factor I: sterile water without fructose, concentration 10^6 conidia/ml without fructose, concentration 10^8 conidia/ml without fructose, concentration 10^10 conidia/ml: without fructose, sterile water + fructose 5 mg/ml, concentration 10^6 conidia/ml without fructose 5 mg/ml, concentration 10^8 conidia/ml + fructose (fungi application before insect infestation and W1 (fungi application after insect infestation. Research phase II was carried out with the best treatment combination compare with control treatment in Temanggung and Bantul. The results of experiment showed that the initial die of D. citri caused by M. anisopliae infection are on 4-6 days after application. The application of M. anisopliae at concentration 10^10 conidia/ml without fructose, applied after insect infestation was most effective. The application after insect infestation was more effective compare with application before insect infestation especially on 4 days after application, but on 35th days after application there was no significant difference. Fructose has no effect to mortality of D. citri.

Diaphorina citri Induces Huanglongbing-Infected Citrus Plant Volatiles to Repel and Reduce the Performance of Propylaea japonica.

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Lin, Yongwen; Lin, Sheng; Akutse, Komivi S; Hussain, Mubasher; Wang, Liande

2016-01-01

Transmission of plant pathogens through insect vectors is a complex biological process involving interactions between the host plants, insects, and pathogens. Simultaneous impact of the insect damage and pathogenic bacteria in infected host plants induce volatiles that modify not only the behavior of its insect vector but also of their natural enemies, such as parasitoid wasps. Therefore, it is essential to understand how insects such as the predator ladybird beetle responds to volatiles emitted from a host plant and how the disease transmission alters the interactions between predators, vector, pathogens, and plants. In this study, we investigated the response of Propylaea japonica to volatiles from citrus plants damaged by Diaphorina citri and Candidatus Liberibacter asiaticus through olfactometer bioassays. Synthetic chemical blends were also used to determine the active compounds in the plant volatile. The results showed that volatiles emitted by healthy plants attracted more P. japonica than other treatments, due to the presence of high quantities of D-limonene and beta-ocimene, and the lack of methyl salicylate. When using synthetic chemicals in the olfactory tests, we found that D-limonene attracted P. japonica while methyl salicylate repelled the predator. However, beta-ocimene attracted the insects at lower concentrations but repelled them at higher concentrations. These results indicate that P. japonica could not efficiently search for its host by using volatile cues emitted from psyllids- and Las bacteria-infected citrus plants.

Impacts of Horticultural Mineral Oils and Two Insecticide Practices on Population Fluctuation of Diaphorina citri and Spread of Huanglongbing in a Citrus Orchard in Sarawak

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Stephen Chan Teck Leong

2012-01-01

Full Text Available Aspects of the incidence and spread of the citrus disease huanglongbing (HLB in relation to the vector Diaphorina citri population fluctuation were studied from January 1999 to December 2001 seasons in a 0.8 ha citrus orchard at Jemukan (1° 33′N, 110° 41′E, Southwest Sarawak in Malaysia. In relation to insecticide and horticultural mineral oils (HMOs use, levels of HLB infection rose quite rapidly over the next 3 years in the unsprayed control and less rapidly in the other treatments such as imidacloprid, nC24HMO, and triazophos/cypermethrin/chlorpyrifos. Levels of HLB as determined by Polymerase Chain Reaction (PCR were 42.2%, 9.4%, 11.4%, and 22.7%, respectively. The effects of nC24HMO and conventional pesticides on the citrus psyllid population and parasitoids in citrus orchard were also determined.

Infection Density Dynamics of the Citrus Greening Bacterium "Candidatus Liberibacter asiaticus" in Field Populations of the Psyllid Diaphorina citri and Its Relevance to the Efficiency of Pathogen Transmission to Citrus Plants.

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Ukuda-Hosokawa, Rie; Sadoyama, Yasutsune; Kishaba, Misaki; Kuriwada, Takashi; Anbutsu, Hisashi; Fukatsu, Takema

2015-06-01

Huanglongbing, or citrus greening, is a devastating disease of citrus plants recently spreading worldwide, which is caused by an uncultivable bacterial pathogen, "Candidatus Liberibacter asiaticus," and vectored by a phloem-sucking insect, Diaphorina citri. We investigated the infection density dynamics of "Ca. Liberibacter asiaticus" in field populations of D. citri with experiments using field-collected insects to address how "Ca. Liberibacter asiaticus" infection density in the vector insect is relevant to pathogen transmission to citrus plants. Of 500 insects continuously collected from "Ca. Liberibacter asiaticus"-infected citrus trees with pathological symptoms in the spring and autumn of 2009, 497 (99.4%) were "Ca. Liberibacter asiaticus" positive. The infections were systemic across head-thorax and abdomen, ranging from 10(3) to 10(7) bacteria per insect. In spring, the infection densities were low in March, at ∼ 10(3) bacteria per insect, increasing up to 10(6) to 10(7) bacteria per insect in April and May, and decreasing to 10(5) to 10(6) bacteria per insect in late May, whereas the infection densities were constantly ∼ 10(6) to 10(7) bacteria per insect in autumn. Statistical analysis suggested that several factors, such as insect sex, host trees, and collection dates, may be correlated with "Ca. Liberibacter asiaticus" infection densities in field D. citri populations. Inoculation experiments with citrus seedlings using field-collected "Ca. Liberibacter asiaticus"-infected insects suggested that (i) "Ca. Liberibacter asiaticus"-transmitting insects tend to exhibit higher infection densities than do nontransmitting insects, (ii) a threshold level (∼ 10(6) bacteria per insect) of "Ca. Liberibacter asiaticus" density in D. citri is required for successful transmission to citrus plants, and (iii) D. citri attaining the threshold infection level transmits "Ca. Liberibacter asiaticus" to citrus plants in a stochastic manner. These findings provide

Oral delivery of double-stranded RNAs induces mortality in nymphs and adults of the Asian citrus psyllid, Diaphorina citri.

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Galdeano, Diogo Manzano; Breton, Michèle Claire; Lopes, João Roberto Spotti; Falk, Bryce W; Machado, Marcos Antonio

2017-01-01

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is one of the most important citrus pests. ACP is the vector of the phloem-limited bacteria Candidatus Liberibacter americanus and Candidatus Liberibacter asiaticus, the causal agents of the devastating citrus disease huanglongbing (HLB). The management of HLB is based on the use of healthy young plants, eradication of infected plants and chemical control of the vector. RNA interference (RNAi) has proven to be a promising tool to control pests and explore gene functions. Recently, studies have reported that target mRNA knockdown in many insects can be induced through feeding with double-stranded RNA (dsRNA). In the current study, we targeted the cathepsin D, chitin synthase and inhibitor of apoptosis genes of adult and nymph ACP by feeding artificial diets mixed with dsRNAs and Murraya paniculata leaves placed in dsRNAs solutions, respectively. Adult ACP mortality was positively correlated with the amount of dsRNA used. Both nymphs and adult ACP fed dsRNAs exhibited significantly increased mortality over time compared with that of the controls. Moreover, qRT-PCR analysis confirmed the dsRNA-mediated RNAi effects on target mRNAs. These results showed that RNAi can be a powerful tool for gene function studies in ACP and perhaps for HLB control.

Analyses of Mitogenome Sequences Revealed that Asian Citrus Psyllids (Diaphorina citri) from California Were Related to Those from Florida.

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Wu, Fengnian; Kumagai, Luci; Cen, Yijing; Chen, Jianchi; Wallis, Christopher M; Polek, MaryLou; Jiang, Hongyan; Zheng, Zheng; Liang, Guangwen; Deng, Xiaoling

2017-08-31

Asian citrus psyllid (ACP, Diaphorina citri Kuwayama) transmits "Candidatus Liberibacter asiaticus" (CLas), an unculturable alpha-proteobacterium associated with citrus Huanglongbing (HLB). CLas has recently been found in California. Understanding ACP population diversity is necessary for HLB regulatory practices aimed at reducing CLas spread. In this study, two circular ACP mitogenome sequences from California (mt-CApsy, ~15,027 bp) and Florida (mt-FLpsy, ~15,012 bp), USA, were acquired. Each mitogenome contained 13 protein coding genes, 2 ribosomal RNA and 22 transfer RNA genes, and a control region varying in sizes. The Californian mt-CApsy was identical to the Floridian mt-FLpsy, but different from the mitogenome (mt-GDpsy) of Guangdong, China, in 50 single nucleotide polymorphisms (SNPs). Further analyses were performed on sequences in cox1 and trnAsn regions with 100 ACPs, SNPs in nad1-nad4-nad5 locus through PCR with 252 ACP samples. All results showed the presence of a Chinese ACP cluster (CAC) and an American ACP cluster (AAC). We proposed that ACP in California was likely not introduced from China based on our current ACP collection but somewhere in America. However, more studies with ACP samples from around the world are needed. ACP mitogenome sequence analyses will facilitate ACP population research.

Occurrence of coccinellids that prey on Diaphorinia citri (Hemiptera:Liviidae) on Euphorbia heterophylla (Euphorbiacea) and Chamaecrista fasciculata (Fabaceae) in a south Florida residential area

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Adding plant-based nutritional resources to a landscape may help to recruit and retain predaceous and parasitic arthropods. As a first step in determining whether the addition of nectary plants can increase the abundance and diversity of biocontrol agents of Diaphorina citri and psyllid suppression ...

Oral delivery of double-stranded RNAs induces mortality in nymphs and adults of the Asian citrus psyllid, Diaphorina citri.

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Diogo Manzano Galdeano

Full Text Available The Asian citrus psyllid (ACP, Diaphorina citri Kuwayama, is one of the most important citrus pests. ACP is the vector of the phloem-limited bacteria Candidatus Liberibacter americanus and Candidatus Liberibacter asiaticus, the causal agents of the devastating citrus disease huanglongbing (HLB. The management of HLB is based on the use of healthy young plants, eradication of infected plants and chemical control of the vector. RNA interference (RNAi has proven to be a promising tool to control pests and explore gene functions. Recently, studies have reported that target mRNA knockdown in many insects can be induced through feeding with double-stranded RNA (dsRNA. In the current study, we targeted the cathepsin D, chitin synthase and inhibitor of apoptosis genes of adult and nymph ACP by feeding artificial diets mixed with dsRNAs and Murraya paniculata leaves placed in dsRNAs solutions, respectively. Adult ACP mortality was positively correlated with the amount of dsRNA used. Both nymphs and adult ACP fed dsRNAs exhibited significantly increased mortality over time compared with that of the controls. Moreover, qRT-PCR analysis confirmed the dsRNA-mediated RNAi effects on target mRNAs. These results showed that RNAi can be a powerful tool for gene function studies in ACP and perhaps for HLB control.

Localization and dynamics of Wolbachia infection in Asian citrus psyllid Diaphorina citri, the insect vector of the causal pathogens of Huanglongbing.

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Ren, Su-Li; Li, Yi-Han; Ou, Da; Guo, Yan-Jun; Qureshi, Jawwad A; Stansly, Philip A; Qiu, Bao-Li

2018-03-23

Wolbachia is a group of intracellular bacteria that infect a wide range of arthropods including the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama. This insect is the vector of Candidatus Liberibacter asiaticus (CLas), the causal pathogen of Huanglongbing or citrus greening disease. Here, we investigated the localization pattern and infection dynamics of Wolbachia in different developmental stages of ACP. Results revealed that all developmental stages of ACP including egg, 1st-5th instar nymphs, and adults of both gender were infected with Wolbachia. FISH visualization of an ACP egg showed that Wolbachia moved from the egg stalk of newly laid eggs to a randomly distributed pattern throughout the egg prior to hatching. The infection rate varied between nymphal instars. The titers of Wolbachia in fourth and fifth instar nymphs were significantly higher than those in the first and second instar nymphs. Wolbachia were scattered in all nymphal stages, but with highest intensity in the U-shaped bacteriome located in the abdomen of the nymph. Wolbachia was confined to two symmetrical organizations in the abdomen of newly emerged female and male adults. The potential mechanisms of Wolbachia infection dynamics are discussed. © 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

INSECTICIDAS DE BAJO IMPACTO AMBIENTAL PARA EL CONTROL DE Diaphorina citri (HEMIPTERA:PSYLLIDAE EN LIMÓN PERSA EN “LA FORTUNA”, NAYARIT, MÉXICO

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Isiordia-Aquino N, Ortiz-Catón M

2013-07-01

Full Text Available El psílido asiático Diaphorina citri es transmisor de la principal enfermedad de los cítricos (huanglongbing, causada por la bacteria Candidatus Liberibacter spp., en este contexto se realizó el presente estudio con la finalidad de evaluar la eficacia de thiacloprid (0.2 y 0.3 L•ha-1, abamectin (0.6 y 1 L•ha-1, aceite mineral (1 y 2 % y sales de ácidos grasos (0.75 y 1 % en ninfas del psílido, en una huerta de limón persa en “La Fortuna” Tepic, Nayarit, México. Se encontró que solamente el tratamiento de abamectin a dosis de 1 L•ha-1 mostró diferencias a las dos semanas después de la primera aplicación y una semana después de la segunda aplicación, con una media de tres ninfas por brote y 93.1 % de mortalidad en comparación al muestreo inicial. Sin embargo, los tratamientos: aceite mineral (1 y 2 %, sales de ácidos grasos (0.75 y 1 % y abamectin (0.6 L•ha-1 mostraron porcentajes de control de 75 a 90 %, además de que se observó la presencia de enemigos naturales del psílido en los tratamientos de aceite mineral (1 y 2 %, sales de ácidos grasos (0.75 y 1 % y el testigo, lo que pudo haber contribuido a la disminución del 44.4 % en este último tratamiento. Con base en estos resultados, impacto ambiental y el costo de los tratamientos para implementarlos, se concluye que los tratamientos de aceite mineral (1 y 2 % y sales de ácidos grasos (0.75 y 1 % son los que presentaron mayor control de ninfas de D. citri y menor costo de aplicación.

76 FR 4859 - Availability of an Environmental Assessment and Finding of No Significant Impact for a Biological...

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2011-01-27

... (Diaphorina citri Kuwayama). The environmental assessment considers the effects of, and alternatives to, the...) for the Biological Control of Asian Citrus Psyllid (Diaphorina citri Kuwayama) in the Continental...

Metabolic alterations in the nymphal instars of Diaphorina citri induced by Candidatus Liberibacter asiaticus, the putative pathogen of huanglongbing.

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Nabil Killiny

Full Text Available Currently, huanglongbing is the most damaging disease of citrus causing huge economic losses. The disease is caused by the Gram-negative bacterium Candidatus Liberibacter asiaticus (CLas. The pathogen is transmitted in a persistent propagative circulative manner within its vector, the Asian citrus psyllid, Diaphorina citri. Exploring the metabolic alteration in the vector may lead to a better understanding of the nutritional needs of CLas and to designing an artificial medium for culturing the pathogen. It has been shown that the nymphal stages have a greater role in transmission mainly because they feed on plants more actively than adults. In this study, we carried out an untargeted comparative metabolomic analysis for healthy and CLas-infected 4th / 5th instar nymphs. The metabolic analysis was performed using trimethylsilylation and methyl chloroformate derivatization followed by Gas Chromatography-Mass Spectrometry (GC-MS. Overall, the changes in the nymph metabolism due to the infection with CLas were more pronounced than in adults, as we previously published. Nymphs reared on CLas-infected Valencia sweet orange were higher in many metabolites, mainly those of the TCA cycle, C16 and C18 fatty acids, glucose, sucrose, L-proline, L-serine, pyroglutamic acid, saccharic acid, threonic acid and myo-inositol than those reared on healthy plants. In contrast, CLas-infected nymphs were lower in putrescine, glycine, L -phenylalanine, L -tyrosine, L -valine, and chiro-inositol. The information provided from this study may contribute in acceleration of the availability of CLas in culture and consequent screening of antibacterial compounds to discover a definitive solution for huanglongbing.

Metabolic alterations in the nymphal instars of Diaphorina citri induced by Candidatus Liberibacter asiaticus, the putative pathogen of huanglongbing.

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Killiny, Nabil; Jones, Shelley E

2018-01-01

Currently, huanglongbing is the most damaging disease of citrus causing huge economic losses. The disease is caused by the Gram-negative bacterium Candidatus Liberibacter asiaticus (CLas). The pathogen is transmitted in a persistent propagative circulative manner within its vector, the Asian citrus psyllid, Diaphorina citri. Exploring the metabolic alteration in the vector may lead to a better understanding of the nutritional needs of CLas and to designing an artificial medium for culturing the pathogen. It has been shown that the nymphal stages have a greater role in transmission mainly because they feed on plants more actively than adults. In this study, we carried out an untargeted comparative metabolomic analysis for healthy and CLas-infected 4th / 5th instar nymphs. The metabolic analysis was performed using trimethylsilylation and methyl chloroformate derivatization followed by Gas Chromatography-Mass Spectrometry (GC-MS). Overall, the changes in the nymph metabolism due to the infection with CLas were more pronounced than in adults, as we previously published. Nymphs reared on CLas-infected Valencia sweet orange were higher in many metabolites, mainly those of the TCA cycle, C16 and C18 fatty acids, glucose, sucrose, L-proline, L-serine, pyroglutamic acid, saccharic acid, threonic acid and myo-inositol than those reared on healthy plants. In contrast, CLas-infected nymphs were lower in putrescine, glycine, L -phenylalanine, L -tyrosine, L -valine, and chiro-inositol. The information provided from this study may contribute in acceleration of the availability of CLas in culture and consequent screening of antibacterial compounds to discover a definitive solution for huanglongbing.

Impact of different temperatures on survival and energy metabolism in the Asian citrus psyllid, Diaphorina citri Kuwayama.

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El-Shesheny, Ibrahim; Hijaz, Faraj; El-Hawary, Ibrahim; Mesbah, Ibrahim; Killiny, Nabil

2016-02-01

Temperature influences the life history and metabolic parameters of insects. Asian citrus psyllid (ACP), Diaphorina citri is a tropical and subtropical pest. ACP invaded new regions around the world and threatened the citrus industry as a vector for Huanglongbing (HLB) disease. ACP is widely distributed and can survive high (up to 45 °C) and low temperatures (as low as -6 °C). The precise mechanism of temperature tolerance in ACP is poorly understood. We investigated adult survival, cellular energy balance, gene expression, and nucleotide and sugar-nucleotide changes under the effect of different temperature regimes (0 °C to 45 °C with 5 °C intervals). The optimum temperatures for survival were 20 and 25 °C. Low temperatures of 0 °C and 5 °C caused 50% mortality after 2 and 4 days respectively, while one day at high temperature (40 °C and 45 °C) caused more than 95% mortality. The lowest quantity of ATP (3.69 ± 1.6 ng/insect) and the maximum ATPase enzyme activities (57.43 ± 7.6 μU/insect) were observed at 25 °C. Correlation between ATP quantities and ATPase activity was negative. Gene expression of hsp 70, V-type proton ATPase catalytic subunit A and ATP synthase α subunit matched these results. Twenty-four nucleotides and sugar-nucleotides were quantified using HPLC in ACP adults maintained at low, high, and optimum temperatures. The nucleotide profiles were different among treatments. The ratios between AMP:ATP and ADP:ATP were significantly decreased and positively correlated to adults survival, whereas the adenylate energy charge was increased in response to low and high temperatures. Exploring energy metabolic regulation in relation with adult survival might help in understanding the physiological basis of how ACP tolerates newly invaded regions. Copyright © 2015 Elsevier Inc. All rights reserved.

Oviposition Behavior and Survival of Tamarixia radiata (Hymenoptera: Eulophidae), an Ectoparasitoid of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae), on Hosts Exposed to an Entomopathogenic Fungus, Isaria fumosorosea (Hypocreales: Cordycipitaceae), Under Laboratory Conditions.

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Chow, Andrew; Dunlap, Christopher A; Jackson, Mark A; Flores, Daniel; Patt, Joseph M; Sétamou, Mamoudou

2016-10-01

Antagonistic interactions between the nymphal parasitoid, Tamarixia radiata Waterston (Hymenoptera: Eulophidae), and the ARSEF 3581 strain of the entomopathogenic fungus, Isaria fumosorosea Wize (Hypocreales: Cordycipitaceae), could disrupt biological control of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). Three interactions were evaluated under laboratory conditions at 25 °C: 1) parasitoid survival if parasitized hosts were exposed to ARSEF 3581 blastospores before or after host mummification; 2) parasitoid survival if mummies containing larva or pupa were exposed to ARSEF 3581 hyphae; 3) parasitoid oviposition on infected hosts with visible or without visible hyphae. Topical application of blastospore formulation onto the dorsal surfaces of live nymphs parasitized with second-instar wasp larva (3 d after parasitism) reduced host mummification by 50% and parasitoid emergence by 85%. However, parasitoid emergence was not affected by topical application of blastospore formulation onto mummies that contained fourth-instar wasp larva (6 d after parasitism). Parasitoid emergence was reduced by 80% if mummies containing fourth-instar wasp larva were covered with blastospore formulation colonized by fungal hyphae. In comparison, parasitoid emergence was not affected if mummies containing wasp pupa (9 d after parasitism) were covered with formulation colonized by fungal hyphae. Female parasitoids oviposited on infected hosts without visible hyphae but not on infected hosts with visible hyphae. Our findings suggest that I. fumosorosea could detrimentally affect T. radiata, if both natural enemies are simultaneously deployed for biological control of D. citri However, temporal separation of the fungus and parasitoid could reduce antagonism and enhance control of D. citri. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Exogenous application of the plant signalers methyl jasmonate and salicylic acid induces changes in volatile emissions from citrus foliage and influences the aggregation behavior of Asian citrus psyllid (Diaphorina citri, vector of Huanglongbing.

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Joseph M Patt

Full Text Available Huanglongbing, also known as citrus greening, is a destructive disease that threatens citrus production worldwide. It is putatively caused by the phloem-limited bacterium Candidatus Liberibacter asiaticus (Las. Currently, the disease is untreatable and efforts focus on intensive insecticide use to control the vector, Asian citrus psyllid (Diaphorina citri. Emerging psyllid resistance to multiple insecticides has generated investigations into the use of exogenously applied signaling compounds to enhance citrus resistance to D. citri and Las. In the present study, we examined whether foliar applications of methyl jasmonate (MJ, a volatile signaling compound associated with the induced systemic resistance pathway, and salicylic acid, a constituent of the systemic acquired resistance pathway, would elicit the emission of defense-related volatiles in citrus foliage, and what effect this might have on the host-plant searching behavior of D. citri. Comparisons were made of volatiles emitted from growing shoots of uninfected and Las-infected 'Valencia' sweet orange (Citrus sinensis trees over two consecutive sampling days. A settling behavioral assay was used to compare psyllid attraction to MJ-treated vs. Tween-treated citrus sprigs. All three main effects, Las infection status, plant signaler application, and sampling day, influenced the proportions of individual volatile compounds emitted in different treatment groups. MJ- and SA-treated trees had higher emission rates than Tween-treated trees. Methyl salicylate (MeSA and β-caryophyllene were present in higher proportions in the volatiles collected from Las-infected + trees. On the other hand, Las-infected + MJ-treated trees emitted lower proportions of MeSA than did Las-infected + Tween-treated trees. Because MeSA is a key D. citri attractant, this result suggests that MJ application could suppress MeSA emission from Las-infected trees, an approach that could be used to discourage psyllid

Exogenous application of the plant signalers methyl jasmonate and salicylic acid induces changes in volatile emissions from citrus foliage and influences the aggregation behavior of Asian citrus psyllid (Diaphorina citri), vector of Huanglongbing.

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Patt, Joseph M; Robbins, Paul S; Niedz, Randy; McCollum, Greg; Alessandro, Rocco

2018-01-01

Huanglongbing, also known as citrus greening, is a destructive disease that threatens citrus production worldwide. It is putatively caused by the phloem-limited bacterium Candidatus Liberibacter asiaticus (Las). Currently, the disease is untreatable and efforts focus on intensive insecticide use to control the vector, Asian citrus psyllid (Diaphorina citri). Emerging psyllid resistance to multiple insecticides has generated investigations into the use of exogenously applied signaling compounds to enhance citrus resistance to D. citri and Las. In the present study, we examined whether foliar applications of methyl jasmonate (MJ), a volatile signaling compound associated with the induced systemic resistance pathway, and salicylic acid, a constituent of the systemic acquired resistance pathway, would elicit the emission of defense-related volatiles in citrus foliage, and what effect this might have on the host-plant searching behavior of D. citri. Comparisons were made of volatiles emitted from growing shoots of uninfected and Las-infected 'Valencia' sweet orange (Citrus sinensis) trees over two consecutive sampling days. A settling behavioral assay was used to compare psyllid attraction to MJ-treated vs. Tween-treated citrus sprigs. All three main effects, Las infection status, plant signaler application, and sampling day, influenced the proportions of individual volatile compounds emitted in different treatment groups. MJ- and SA-treated trees had higher emission rates than Tween-treated trees. Methyl salicylate (MeSA) and β-caryophyllene were present in higher proportions in the volatiles collected from Las-infected + trees. On the other hand, Las-infected + MJ-treated trees emitted lower proportions of MeSA than did Las-infected + Tween-treated trees. Because MeSA is a key D. citri attractant, this result suggests that MJ application could suppress MeSA emission from Las-infected trees, an approach that could be used to discourage psyllid colonization during

Exogenous application of the plant signalers methyl jasmonate and salicylic acid induces changes in volatile emissions from citrus foliage and influences the aggregation behavior of Asian citrus psyllid (Diaphorina citri), vector of Huanglongbing

Science.gov (United States)

Robbins, Paul S.; Niedz, Randy; McCollum, Greg; Alessandro, Rocco

2018-01-01

Huanglongbing, also known as citrus greening, is a destructive disease that threatens citrus production worldwide. It is putatively caused by the phloem-limited bacterium Candidatus Liberibacter asiaticus (Las). Currently, the disease is untreatable and efforts focus on intensive insecticide use to control the vector, Asian citrus psyllid (Diaphorina citri). Emerging psyllid resistance to multiple insecticides has generated investigations into the use of exogenously applied signaling compounds to enhance citrus resistance to D. citri and Las. In the present study, we examined whether foliar applications of methyl jasmonate (MJ), a volatile signaling compound associated with the induced systemic resistance pathway, and salicylic acid, a constituent of the systemic acquired resistance pathway, would elicit the emission of defense-related volatiles in citrus foliage, and what effect this might have on the host-plant searching behavior of D. citri. Comparisons were made of volatiles emitted from growing shoots of uninfected and Las-infected ‘Valencia’ sweet orange (Citrus sinensis) trees over two consecutive sampling days. A settling behavioral assay was used to compare psyllid attraction to MJ-treated vs. Tween-treated citrus sprigs. All three main effects, Las infection status, plant signaler application, and sampling day, influenced the proportions of individual volatile compounds emitted in different treatment groups. MJ- and SA-treated trees had higher emission rates than Tween-treated trees. Methyl salicylate (MeSA) and β-caryophyllene were present in higher proportions in the volatiles collected from Las-infected + trees. On the other hand, Las-infected + MJ-treated trees emitted lower proportions of MeSA than did Las-infected + Tween-treated trees. Because MeSA is a key D. citri attractant, this result suggests that MJ application could suppress MeSA emission from Las-infected trees, an approach that could be used to discourage psyllid colonization during

Xanthomonas citri pv. citri pathotypes

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Di Lorenzo, Flaviana; Silipo, Alba; Gersby, Lotte Bettina Andersen

2017-01-01

Xanthomonas citri pv. citri is the pathogen responsible for Asiatic citrus canker, one of the most serious citrus diseases worldwide. The lipopolysaccharide (LPS) molecule has been demonstrated to be involved in X. citri pv. citri virulence. Despite enormous progress in investigations of the mole...

Global genetic variation in the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae) and the endosymbiont Wolbachia: links between Iran and the USA detected.

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Lashkari, Mohammadreza; Manzari, Shahab; Sahragard, Ahad; Malagnini, Valeria; Boykin, Laura M; Hosseini, Reza

2014-07-01

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is one of the most serious pests of citrus in the world, because it transmits the pathogen that causes citrus greening disease. To determine genetic variation among geographic populations of D. citri, microsatellite markers, mitochondrial gene cytochrome oxidase I (mtCOI) and the Wolbachia-Diaphorina, wDi, gene wsp sequence data were used to characterize Iranian and Pakistani populations. Also, a Bayesian phylogenetic technique was utilized to elucidate the relationships among the sequences data in this study and all mtCOI and wsp sequence data available in GenBank and the Wolbachia database. Microsatellite markers revealed significant genetic differentiation among Iranian populations, as well as between Iranian and Pakistani populations (FST  = 0.0428, p citri populations in Iran, India, Saudi Arabia, Brazil, Mexico, Florida and Texas (USA) are similar. Wolbachia, wDi, wsp sequences were similar among Iranian populations, but different between Iranian and Pakistani populations. The South West Asia (SWA) group is the most likely source of the introduced Iranian populations of D. citri. This assertion is also supported by the sequence similarity of the Wolbachia, wDi, strains from the Florida, USA and Iranian D. citri. These results should be considered when looking for biological controls in either country. © 2013 Society of Chemical Industry.

Essential Oil Variation from Twenty Two Genotypes of Citrus in Brazil-Chemometric Approach and Repellency Against Diaphorina citri Kuwayama.

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Andrade, Moacir Dos Santos; Ribeiro, Leandro do Prado; Borgoni, Paulo Cesar; Silva, Maria Fátima das Graças Fernandes da; Forim, Moacir Rossi; Fernandes, João Batista; Vieira, Paulo Cezar; Vendramin, José Djair; Machado, Marcos Antônio

2016-06-22

The chemical composition of volatile oils from 22 genotypes of Citrus and related genera was poorly differentiated, but chemometric techniques have clarified the relationships between the 22 genotypes, and allowed us to understand their resistance to D. citri. The most convincing similarities include the synthesis of (Z)-β-ocimene and (E)-caryophyllene for all 11 genotypes of group A. Genotypes of group B are not uniformly characterized by essential oil compounds. When stimulated with odor sources of 22 genotypes in a Y-tube olfactometer D. citri preferentially entered the arm containing the volatile oils of Murraya paniculata, confirming orange jasmine as its best host. C. reticulata × C. sinensis was the least preferred genotype, and is characterized by the presence of phytol, (Z)-β-ocimene, and β-elemene, which were not found in the most preferred genotype. We speculate that these three compounds may act as a repellent, making these oils less attractive to D. citri.

Essential Oil Variation from Twenty Two Genotypes of Citrus in Brazil—Chemometric Approach and Repellency Against Diaphorina citri Kuwayama

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Moacir dos Santos Andrade

2016-06-01

Full Text Available The chemical composition of volatile oils from 22 genotypes of Citrus and related genera was poorly differentiated, but chemometric techniques have clarified the relationships between the 22 genotypes, and allowed us to understand their resistance to D. citri. The most convincing similarities include the synthesis of (Z-β-ocimene and (E-caryophyllene for all 11 genotypes of group A. Genotypes of group B are not uniformly characterized by essential oil compounds. When stimulated with odor sources of 22 genotypes in a Y-tube olfactometer D. citri preferentially entered the arm containing the volatile oils of Murraya paniculata, confirming orange jasmine as its best host. C. reticulata × C. sinensis was the least preferred genotype, and is characterized by the presence of phytol, (Z-β-ocimene, and β-elemene, which were not found in the most preferred genotype. We speculate that these three compounds may act as a repellent, making these oils less attractive to D. citri.

'Psyllid purple’: evidence of behavior-based utilization by the Asian citrus psyllid of a combination of short and long wavelengths

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The Asian citrus psyllid, Diaphorina citri, is the vector of huanglongbing, the most serious disease affecting citrus globally. In Florida alone, D. citri has resulted in billions of dollars of damage and has spread to all the citrus growing regions of North America. The visual behavior of D. citri ...

Behavioral, Ultrastructural and Chemical Studies on the Honeydew and Waxy Secretions by Nymphs and Adults of the Asian Citrus Psyllid Diaphorina citri (Hemiptera: Psyllidae)

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Ammar, El-Desouky; Alessandro, Rocco; Shatters Jr, Robert G.; Hall, David G.

2013-01-01

The Asian citrus psyllid (ACP), Diaphorina citri (Hemiptera: Psyllidae) is the primary vector of the bacterium causing citrus huanglongbing (citrus greening), the most serious disease of citrus worldwide. Psyllids and other hemipterans produce large amounts of honeydew, which has been used previously as an indicator of phloem sap composition and insect feeding or metabolism. Behavioral, ultrastructural and chemical studies on ACP, its honeydew and waxy secretions showed important differences between nymphs, males and females, and suggested some mechanisms by which the psyllids, especially nymphs and adult females, can minimize their contamination with honeydew excretions. The anal opening in ACP, near the posterior end of the abdomen, is on the ventral side in nymphs and on the dorsal side in adult males and females. Video recordings showed that adult males produce clear sticky droplets of honeydew gently deposited behind their body on the leaf surface, whereas adult females produce whitish honeydew pellets powerfully propelled away from the female body, probably to get their excretions away from eggs and newly hatched nymphs. ACP nymphs produce long ribbons or tubes of honeydew that frequently stay attached to the exuviae after molting, or drop when feeding on the lower side of citrus leaves. Furthermore, honeydew excretions of both nymphs and adult females are covered with a thin layer of whitish waxy material ultrastructurally composed of a convoluted network of long fine filaments or ribbons. This material is extruded from intricate arrays of wax pores in the circumanal ring (around the anus) that is found in nymphs and females but not in males of ACP or other psyllid species. Infrared microscopy and mass spectroscopy revealed that, in addition to various sugars, honeydew excretions of ACP nymphs and females are covered with a thin layer of wax similar in profile to ester waxes. PMID:23762268

Stylet morphometrics and citrus leaf vein structure in relation to feeding behavior of the Asian citrus psyllid Diaphorina citri, vector of citrus huanglongbing bacterium.

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El-Desouky Ammar

Full Text Available The Asian citrus psyllid (ACP, Diaphorina citri (Hemiptera: Psyllidae, is the primary vector of the phloem-limited bacterium Candidatus Liberibacter asiaticus (LAS associated with huanglongbing (HLB, citrus greening, considered the world's most serious disease of citrus. Stylet morphometrics of ACP nymphs and adults were studied in relation to citrus vein structure and to their putative (histologically verified feeding sites on Valencia orange leaves. ACP nymphs preferred to settle and feed on the lower (abaxial side of young leaves either on secondary veins or on the sides of the midrib, whereas adults preferred to settle and feed on the upper (adaxial or lower secondary veins of young or old leaves. Early instar nymphs can reach and probe the phloem probably because the distance to the phloem is considerably shorter in younger than in mature leaves, and is shorter from the sides of the midrib compared to that from the center. Additionally, the thick-walled 'fibrous ring' (sclerenchyma around the phloem, which may act as a barrier to ACP stylet penetration into the phloem, is more prominent in older than in younger leaves and in the center than on the sides of the midrib. The majority (80-90% of the salivary sheath termini produced by ACP nymphs and adults that reached a vascular bundle were associated with the phloem, whereas only 10-20% were associated with xylem vessels. Ultrastructural studies on ACP stylets and LAS-infected leaves suggested that the width of the maxillary food canal in first instar nymphs is wide enough for LAS bacteria to traverse during food ingestion (and LAS acquisition. However, the width of the maxillary salivary canal in these nymphs may not be wide enough to accommodate LAS bacteria during salivation (and LAS inoculation into host plants. This may explain the inability of early instar nymphs to transmit LAS/HLB in earlier reports.

Behavioral, ultrastructural and chemical studies on the honeydew and waxy secretions by nymphs and adults of the Asian citrus psyllid Diaphorina citri (Hemiptera: Psyllidae.

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El-Desouky Ammar

Full Text Available The Asian citrus psyllid (ACP, Diaphorina citri (Hemiptera: Psyllidae is the primary vector of the bacterium causing citrus huanglongbing (citrus greening, the most serious disease of citrus worldwide. Psyllids and other hemipterans produce large amounts of honeydew, which has been used previously as an indicator of phloem sap composition and insect feeding or metabolism. Behavioral, ultrastructural and chemical studies on ACP, its honeydew and waxy secretions showed important differences between nymphs, males and females, and suggested some mechanisms by which the psyllids, especially nymphs and adult females, can minimize their contamination with honeydew excretions. The anal opening in ACP, near the posterior end of the abdomen, is on the ventral side in nymphs and on the dorsal side in adult males and females. Video recordings showed that adult males produce clear sticky droplets of honeydew gently deposited behind their body on the leaf surface, whereas adult females produce whitish honeydew pellets powerfully propelled away from the female body, probably to get their excretions away from eggs and newly hatched nymphs. ACP nymphs produce long ribbons or tubes of honeydew that frequently stay attached to the exuviae after molting, or drop when feeding on the lower side of citrus leaves. Furthermore, honeydew excretions of both nymphs and adult females are covered with a thin layer of whitish waxy material ultrastructurally composed of a convoluted network of long fine filaments or ribbons. This material is extruded from intricate arrays of wax pores in the circumanal ring (around the anus that is found in nymphs and females but not in males of ACP or other psyllid species. Infrared microscopy and mass spectroscopy revealed that, in addition to various sugars, honeydew excretions of ACP nymphs and females are covered with a thin layer of wax similar in profile to ester waxes.

Metabolic interplay between the Asian citrus psyllid and its Profftella symbiont: An Achilles’ heel of the citrus greening insect vector

Science.gov (United States)

‘Candidatus Liberibacter asiaticus’ (CLas), the bacterial pathogen associated with citrus greening disease, is transmitted by Diaphorina citri, the Asian citrus psyllid. Interactions among D. citri and its microbial endosymbionts, including ‘Candidatus Profftella armatura’, are likely to impact tra...

Primer Registro de Tamarixia radiata (Waterston, 1922 (Hymenoptera: Eulophidae en Colombia Record of Tamarixia radiata (Waterston, 1922 (Hymenoptera: Eulophidae in Colombia

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Everth Emilio Ebratt Ravelo

2011-12-01

Full Text Available Tamarixia radiata es un ectoparasitoide idiobionte de Diaphorina citri Kuwayama (Hemiptera: Psyllidae; la especie fue descrita a partir de material recogido en la India. Ha sido introducida en muchas regiones intencional o accidentalmente para el control biológico de D. citri, incluyendo algunos países asiáticos. Se sabe de su presencia en el sur de África y las Américas. En Suramérica, se reportó en Brasil, Argentina y ahora en Colombia.Tamarixia radiata is an ectoparasitoid idiobionte of Diaphorina citri Kuwayama (Hemiptera: Psyllidae; the species was described from material collected in India. It has been introduced in many regions intentionally or accidentally, to biological control of D. citri, including some Asian countries. It is know about its presence in southern Africa and the Americas. In South America, it is present in Brazil, Argentina and now in Colombia.

Identification of an Extracellular Endoglucanase That Is Required for Full Virulence in Xanthomonas citri subsp. citri.

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Tian Xia

Full Text Available Xanthomonas citri subsp. citri causes citrus canker disease, which is characterized by the formation of water-soaked lesions, white or yellow spongy pustules and brown corky canker. In this work, we report the contribution of extracellular endoglucanase to canker development during infection. The ectopic expression of nine putative cellulases in Escherichia coli indicated that two endoglucanases, BglC3 and EngXCA, show carboxymethyl cellulase activity. Both bglC3 and engXCA genes were transcribed in X. citri subsp. citri, however, only BglC3 protein was detected outside the cell in western blot analysis. The deletion of bglC3 gene resulted in complete loss of extracellular carboxymethyl cellulase activity and delayed the onset of canker symptoms in both infiltration- and wound-inoculation assays. When growing in plant tissue, the cell density of bglC3 mutant was lower than that of the wild type. Our data demonstrated that BglC3 is an extracellular endoglucanase required for the full virulence of X. citri subsp. citri.

Differences in Stylet Sheath Occurrence and the Fibrous Ring (Sclerenchyma) between xCitroncirus Plants Relatively Resistant or Susceptible to Adults of the Asian Citrus Psyllid Diaphorina citri (Hemiptera: Liviidae)

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Ammar, El-Desouky; Richardson, Matthew L.; Abdo, Zaid; Hall, David G.; Shatters, Robert G.

2014-01-01

The Asian citrus psyllid (ACP, Diaphorina citri, Hemiptera: Liviidae), is the principal vector of the phloem-limited bacteria strongly associated with huanglongbing (HLB), the world’s most serious disease of citrus. Host plant resistance may provide an environmentally safe and sustainable method of controlling ACP and/or HLB. Two xCitroncirus accessions (hybrids of Poncirus trifoliata and Citrus spp.), that are relatively resistant (UN-3881) or relatively susceptible (Troyer-1459) to ACP adults with regard to adult longevity, were compared in relation to ACP feeding behavior and some structural features of the leaf midrib. The settling (putative feeding/probing) sites of ACP adults on various parts of the leaf were not influenced primarily by plant accession. However, fewer ACP stylet sheaths were found in the midrib and fewer stylet sheath termini reached the vascular bundle (phloem and/or xylem) in UN-3881 compared to Troyer-1459 plants. Furthermore, in midribs of UN-3881 leaves the fibrous ring (sclerenchyma) around the phloem was significantly wider (thicker) compared to that in midribs of Troyer-1459 leaves. Our data indicate that feeding and/or probing by ACP adults into the vascular bundle is less frequent in the more resistant (UN-3881) than in the more susceptible (Troyer-1459) accessions. Our results also suggest that the thickness of the fibrous ring may be a barrier to stylet penetration into the vascular bundle, which is important for successful ACP feeding on the phloem and for transmitting HLB-associated bacteria. These results may help in the development of citrus plants resistant to ACP, which in turn could halt or slow the spread of the HLB-associated bacteria by this vector. PMID:25343712

Differences in stylet sheath occurrence and the fibrous ring (sclerenchyma between xCitroncirus plants relatively resistant or susceptible to adults of the Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae.

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El-Desouky Ammar

Full Text Available The Asian citrus psyllid (ACP, Diaphorina citri, Hemiptera: Liviidae, is the principal vector of the phloem-limited bacteria strongly associated with huanglongbing (HLB, the world's most serious disease of citrus. Host plant resistance may provide an environmentally safe and sustainable method of controlling ACP and/or HLB. Two xCitroncirus accessions (hybrids of Poncirus trifoliata and Citrus spp., that are relatively resistant (UN-3881 or relatively susceptible (Troyer-1459 to ACP adults with regard to adult longevity, were compared in relation to ACP feeding behavior and some structural features of the leaf midrib. The settling (putative feeding/probing sites of ACP adults on various parts of the leaf were not influenced primarily by plant accession. However, fewer ACP stylet sheaths were found in the midrib and fewer stylet sheath termini reached the vascular bundle (phloem and/or xylem in UN-3881 compared to Troyer-1459 plants. Furthermore, in midribs of UN-3881 leaves the fibrous ring (sclerenchyma around the phloem was significantly wider (thicker compared to that in midribs of Troyer-1459 leaves. Our data indicate that feeding and/or probing by ACP adults into the vascular bundle is less frequent in the more resistant (UN-3881 than in the more susceptible (Troyer-1459 accessions. Our results also suggest that the thickness of the fibrous ring may be a barrier to stylet penetration into the vascular bundle, which is important for successful ACP feeding on the phloem and for transmitting HLB-associated bacteria. These results may help in the development of citrus plants resistant to ACP, which in turn could halt or slow the spread of the HLB-associated bacteria by this vector.

The type III protein secretion system contributes to Xanthomonas citri subsp. citri biofilm formation

KAUST Repository

Zimaro, Tamara

2014-04-18

Background: Several bacterial plant pathogens colonize their hosts through the secretion of effector proteins by a Type III protein secretion system (T3SS). The role of T3SS in bacterial pathogenesis is well established but whether this system is involved in multicellular processes, such as bacterial biofilm formation has not been elucidated. Here, the phytopathogen Xanthomonas citri subsp. citri (X. citri) was used as a model to gain further insights about the role of the T3SS in biofilm formation. Results: The capacity of biofilm formation of different X. citri T3SS mutants was compared to the wild type strain and it was observed that this secretion system was necessary for this process. Moreover, the T3SS mutants adhered proficiently to leaf surfaces but were impaired in leaf-associated growth. A proteomic study of biofilm cells showed that the lack of the T3SS causes changes in the expression of proteins involved in metabolic processes, energy generation, exopolysaccharide (EPS) production and bacterial motility as well as outer membrane proteins. Furthermore, EPS production and bacterial motility were also altered in the T3SS mutants. Conclusions: Our results indicate a novel role for T3SS in X. citri in the modulation of biofilm formation. Since this process increases X. citri virulence, this study reveals new functions of T3SS in pathogenesis. 2014 Zimaro et al.; licensee BioMed Central Ltd.

Expression of Xylella fastidiosa RpfF in citrus disrupts signaling in Xanthomonas citri subsp. citri and thereby its virulence.

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Caserta, R; Picchi, S C; Takita, M A; Tomaz, J P; Pereira, W E L; Machado, M A; Ionescu, M; Lindow, S; De Souza, A A

2014-11-01

Xylella fastidiosa and Xanthomonas citri subsp. citri, that cause citrus variegated chlorosis (CVC) and citrus canker diseases, respectively, utilize diffusible signal factor (DSF) for quorum sensing. DSF, produced by RpfF, are similar fatty acids in both organisms, although a different set of genes is regulated by DSF in each species. Because of this similarity, Xylella fastidiosa DSF might be recognized and affect the biology of Xanthomonas citri. Therefore, transgenic Citrus sinensis and Carrizo citrange plants overexpressing the Xylella fastidiosa rpfF were inoculated with Xanthomonas citri and changes in symptoms of citrus canker were observed. X. citri biofilms formed only at wound sites on transgenic leaves and were thicker; however, bacteria were unable to break through the tissue and form pustules elsewhere. Although abundant growth of X. citri occurred at wound sites on inoculated transgenic leaves, little growth was observed on unwounded tissue. Genes in the DFS-responsive core in X. citri were downregulated in bacteria isolated from transgenic leaves. DSF-dependent expression of engA was suppressed in cells exposed to xylem sap from transgenic plants. Thus, altered symptom development appears to be due to reduced expression of virulence genes because of the presence of antagonists of DSF signaling in X. citri in rpfF-expressing plants.

Oviposition behavior and survival of Tamarixia radiata (Hymenoptera: Eulophidae), an ectoparasitoid of the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae), on hosts exposed to an entomopathogenic fungus

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Antagonistic interactions between the nymphal parasitoid, Tamarixia radiata Waterston (Hymenoptera: Eulophidae), and the ARSEF 3581 isolate of the entomopathogenic fungus, Isaria fumosorosea Wize (Hypocreales: Cordycipitaceae) could disrupt biological control of the Asian citrus psyllid, Diaphorina ...

Metabolomic comparative analysis of the phloem sap of curry leaf tree (Bergera koenegii), orange jasmine (Murraya paniculata), and Valencia sweet orange (Citrus sinensis) supports their differential responses to Huanglongbing

OpenAIRE

Killiny, Nabil

2016-01-01

Orange jasmine, Murraya paniculata and curry leaf tree, Bergera koenegii are alternative hosts for Diaphorina citri, the vector of Candidatus Liberibacter asiaticus (CLas), the pathogen of huanglongbing (HLB) in citrus. D. citri feeds on the phloem sap where CLas grows. It has been shown that orange jasmine was a better host than curry leaf tree to D. citri. In addition, CLas can infect orange jasmine but not curry leaf tree. Here, we compared the phloem sap composition of these 2 plants to t...

Resistance of sweet orange Pera (Citrus sinensis) genotypes to Xanthomonas citri subsp. citri under field conditions

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Citrus canker control is based on protection measures and eradication of plants infected with Xanthomonas citri subsp. citri. Although these measures show satisfactory results, the use of resistant genotypes is an important alternative for citrus canker control. The aim of this study was to evaluate...

A “walker” tool to place Diaphorina citri (Hemiptera: Liviidae) adults at predetermined sites for bioassays of behavior in citrus (Sapindales: Rutacease) trees

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A walker tool was developed to assist placement of D. citri on citrus host trees in behavioral bioassays. The walker performs better than a commonly used paintbrush tool in the proportion of successful placements and in the reduction of jumps away from the citrus leaf, although it takes about two mi...

In Vitro and In Vivo Antibacterial Activity of Some Organic and Inorganic Salts Against Asiatic Citrus Canker Agent Xanthomonas Citri Subsp. Citri

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Vahideh Hasabi

2014-07-01

Full Text Available Asiatic citrus canker caused by Xanthomonas citri subsp. citri is becoming a disease of high economic impact, affecting all types of important citrus crops. In this study, the potential antibacterial activity of ten organic and inorganic salts on X. citri subsp. citri and on citrus canker disease development was evaluated. Among the salt compounds, copper, iron and zinc inorganic salts particularly zinc (with the highest diameter of inhibition, the lowest MIC and MBC values and the highest bacterial growth inhibitory effect had direct antibacterial activity and strongly reduced the development of canker disease and bacterial population of lime plants.

Annotation of the Asian citrus psyllid genome reveals a reduced innate immune system

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Citrus production worldwide is currently facing significant losses due to citrus greening disease, also known as huanglongbing. The citrus greening bacteria, Candidatus Liberibacter asiaticus (CLas), is a persistent propagative pathogen transmitted by the Asian citrus psyllid, Diaphorina citri Kuway...

Combining 'omics and microscopy to visualize interactions between the Asian citrus psyllid vector and the Huanglongbing pathogen Candidatus Liberibacter asiaticus in the insect gut

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Huanglongbing, or citrus greening disease, is an economically devastating bacterial disease of citrus. It is associated with infection by the gram-negative bacterium Candidatus Liberibacter asiaticus (CLas). CLas is transmitted by Diaphorina citri, the Asian citrus psyllid (ACP). For insect transmis...

Improved annotation of the insect vector of citrus greening disease: Biocuration by a diverse genomics community

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The Asian citrus psyllid (Diaphorina citri Kuwayama) is the insect vector of the bacterium Candidatus Liberibacter asiaticus (CLas), the pathogen associated with citrus Huanglongbing (HLB, citrus greening). HLB threatens citrus production worldwide. Suppression or reduction of the insect vector usin...

Ultrastructure of the salivary glands, alimentary canal and bacteria-like organisms in the Asian citrus psyllid, vector of citrus huanglongbing-disease bacteria

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Several psyllids (Hemiptera: Psylloidea) are known as vectors of some economically important viral and bacterial plant pathogens. The Asian citrus psyllid (ACP, Diaphorina citri, Hemiptera, Liviidae) is the principal vector of ‘Candidatus Liberibacter asiaticus’ (Las), the putative bacterial causal ...

Field validation of a system for autodissemination of an entomopathogenic fungus, Isaria fumosorosea, to control the Asian citrus psyllid on residential citrus

Science.gov (United States)

The citrus industries of California and Texas share a pressing problem with the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae) and huanglongbing (HLB) spreading in residential citrus near commercial groves. Insecticidal treatment of residential trees for the psyllid is problem...

The cys regulon of Xanthomonas citri

International Nuclear Information System (INIS)

Moutran, A.; Balan, A.

2012-01-01

Full text: In Escherichia coli, genes involved in metabolic pathway of sulfate and sulfonate compounds are clustered in a cys regulon, which includes three ABC transport system (operons: sbpcysWUA; ssuABC and tauABC), thirteen genes involved in the sulfur reduction (ssuDE; tauD and cysDNCHIJGK) and two regulatory proteins that belong to LysR transcription family: CysB and Cbl. Notably, a search and comparative analysis of these genes in the genomes of the citrus pathogen Xanthomonas citri and other phylogenetically related Xanthomonas species revealed the presence of genes involved with alkanesulfonate, sulfate ester and taurine, only in X. citri, suggesting that proteins from this regulon might be associated with pathogenicity in citrus. Using the molecular modeling associated with a system biology view, we modeled all the protein structures of the X. citri cys regulon as well as characterized the important residues forming the putative active sites. Comparison with orthologs from different microorganisms was made in order to get a phylogenetic relationships. We showed that proteins that are responsible for the affinity and specificity of the alkanesulfonate, sulfate and taurine transport systems conserved the residues involved in the sulfate coordination but are organized in different branches in evolution. Inside these phylogenetic branches, proteins involved in the sulfate transporter are highly conserved when compared to the others. Moreover, we identified that the taurine-binding protein (TauA) of the X. citri belongs to a different evolutionary branch from that one that described for E. coli. These differences were also noticed for components of the tau operon, including a putative new regulator. The function and mechanism of action of each protein is discussed in order to bring light for the sulfur assimilation processes and their importance for X. citri physiology. (author)

The cys regulon of Xanthomonas citri

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Moutran, A.; Balan, A. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil)

2012-07-01

Full text: In Escherichia coli, genes involved in metabolic pathway of sulfate and sulfonate compounds are clustered in a cys regulon, which includes three ABC transport system (operons: sbpcysWUA; ssuABC and tauABC), thirteen genes involved in the sulfur reduction (ssuDE; tauD and cysDNCHIJGK) and two regulatory proteins that belong to LysR transcription family: CysB and Cbl. Notably, a search and comparative analysis of these genes in the genomes of the citrus pathogen Xanthomonas citri and other phylogenetically related Xanthomonas species revealed the presence of genes involved with alkanesulfonate, sulfate ester and taurine, only in X. citri, suggesting that proteins from this regulon might be associated with pathogenicity in citrus. Using the molecular modeling associated with a system biology view, we modeled all the protein structures of the X. citri cys regulon as well as characterized the important residues forming the putative active sites. Comparison with orthologs from different microorganisms was made in order to get a phylogenetic relationships. We showed that proteins that are responsible for the affinity and specificity of the alkanesulfonate, sulfate and taurine transport systems conserved the residues involved in the sulfate coordination but are organized in different branches in evolution. Inside these phylogenetic branches, proteins involved in the sulfate transporter are highly conserved when compared to the others. Moreover, we identified that the taurine-binding protein (TauA) of the X. citri belongs to a different evolutionary branch from that one that described for E. coli. These differences were also noticed for components of the tau operon, including a putative new regulator. The function and mechanism of action of each protein is discussed in order to bring light for the sulfur assimilation processes and their importance for X. citri physiology. (author)

Correlation of electronic monitoring and stylet pathways elucidate the role of sclerenchymatous ring as a barrier to phloem feeding on citrus leaves by Asian citrus psyllid

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Asian citrus psyllid (ACP, Diaphorina. citri) feeding behaviors play a significant role in the transmission of the phloem-limited Candidatus Liberibacter asiaticus (CLas) bacterium that causes the economically devastating citrus greening disease. Recent studies have shown a fibrous ring of thick-wal...

Epifluorescence and stereomicroscopy of trichomes associated with resistant and susceptible host plant genotypes of the Asian citrus psyllid (Hemiptera: Liviidae)

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Epifluorescence, light and stereo-microscopy were used to characterize foliar trichomes associated with young flush leaves and stems of six plant genotypes that are hosts of the Asian citrus psyllid, Diaphorina citri, vector of the economically important citrus greening disease. These genotypes incl...

Factors affecting transmission rates of 'Candidatus Liberibacter asiaticus' by Asian citrus psyllid

Science.gov (United States)

The Asian citrus psyllid (ACP), Diaphorina citri, is an important pest because it transmits a bacterium ‘Candidatus Liberibacter asiaticus’ (CLas) responsible for a serious disease of citrus known as Asiatic huanglongbing (citrus greening disease). USDA-ARS researchers recently established a program...

Degradation products of citrus volatile organic compounds (VOCs) acting as phagostimulants that increase probing behavior of Asian citrus psyllid

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Volatile phytochemicals play a role in orientation by phytophagous insects. We studied antennal and behavioral responses of the Asian citrus psyllid, Diaphorina citri Kuwayama, vector of the citrus greening disease pathogen. Little or no response to citrus leaf volatiles was detected by electroanten...

Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri.

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Zhuo, Tao; Rou, Wei; Song, Xue; Guo, Jing; Fan, Xiaojing; Kamau, Gicharu Gibson; Zou, Huasong

2015-10-23

The carA and carB genes code the small and large subunits of carbamoyl-phosphate synthase (CPS) that responsible for arginine and pyrimidine production. The purpose of this work was to study the gene organization and expression pattern of carAB operon, and the biological functions of carA and carB genes in Xanthomonas citri subsp. citri. RT-PCR method was employed to identify the full length of carAB operon transcript in X. citri subsp. citri. The promoter of carAB operon was predicted and analyzed its activity by fusing a GUS reporter gene. The swimming motility was tested on 0.25% agar NY plates with 1% glucose. Biofilm was measured by cell adhesion to polyvinyl chloride 96-well plate. The results indicated that carAB operon was composed of five gene members carA-orf-carB-greA-rpfE. A single promoter was predicted from the nucleotide sequence upstream of carAB operon, and its sensitivity to glutamic acid, uracil and arginine was confirmed by fusing a GUS reporter gene. Deletion mutagenesis of carB gene resulted in reduced abilities in swimming on soft solid media and in forming biofilm on polystyrene microtiter plates. From these results, we concluded that carAB operon was involved in multiple biological processes in X. citri subsp. citri.

Ultraviolet irradiation mutagenesis and recombination in spiroplasma citri

Energy Technology Data Exchange (ETDEWEB)

Labarere, J.; Barroso, G. (Bordeaux-2 Univ., 33 (France))

1984-09-01

A method for obtaining and screening uv-induced mutants from Spiroplasma citri is described. Lethality response curves showed that S. citri is more sensitive to uv irradiation than are other microorganisms. The presence of a shoulder in the lethality response curve showed the existence of systems able to repair uv-induced DNA damages. Toxic-resistant mutants have been obtained. A uv fluence equal to 10 J/m/sup 2/ multiplied by 2.5x10/sup 3/ gave the spontaneous mutation frequency. Arsenic acid- and xylitol-resistant mutants were used to investigate transfer of genetic information in S. citri. After 90 min of incubation, the recombination frequency was 5x10/sup -5/.

The novel virulence-related gene nlxA in the lipopolysaccharide cluster of Xanthomonas citri ssp. citri is involved in the production of lipopolysaccharide and extracellular polysaccharide, motility, biofilm formation and stress resistance.

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Yan, Qing; Hu, Xiufang; Wang, Nian

2012-10-01

Lipopolysaccharide (LPS) is an important virulence factor of Xanthomonas citri ssp. citri, the causative agent of citrus canker disease. In this research, a novel gene, designated as nlxA (novel LPS cluster gene of X. citri ssp. citri), in the LPS cluster of X. citri ssp. citri 306, was characterized. Our results indicate that nlxA is required for O-polysaccharide biosynthesis by encoding a putative rhamnosyltransferase. This is supported by several lines of evidence: (i) NlxA shares 40.14% identity with WsaF, which acts as a rhamnosyltransferase; (ii) sodium dodecylsulphate-polyacrylamide gel electrophoresis analysis showed that four bands of the O-antigen part of LPS were missing in the LPS production of the nlxA mutant; this is also consistent with a previous report that the O-antigen moiety of LPS of X. citri ssp. citri is composed of a rhamnose homo-oligosaccharide; (iii) mutation of nlxA resulted in a significant reduction in the resistance of X. citri ssp. citri to different stresses, including sodium dodecylsulphate, polymyxin B, H(2)O(2), phenol, CuSO(4) and ZnSO(4). In addition, our results indicate that nlxA plays an important role in extracellular polysaccharide production, biofilm formation, stress resistance, motility on semi-solid plates, virulence and in planta growth in the host plant grapefruit. © 2012 THE AUTHORS. MOLECULAR PLANT PATHOLOGY © 2012 BSPP AND BLACKWELL PUBLISHING LTD.

Asian citrus psyllid RNAi pathway - RNAi evidence

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In silico analyses of the draft genome of Diaphorina citri, the Asian citrus psyllid, for genes within the Ribonucleic acid interference(RNAi), pathway was successful. The psyllid is the vector of the plant-infecting bacterium, Candidatus Liberibacter asiaticus (CLas), which is linked to citrus gree...

Evaluating citrus germplasm for huanglongbing (HLB) resistance: USDA-ARS Inoculation Program

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The Asian citrus psyllid (ACP), Diaphorina citri, is an important pest because it vectors bacteria responsible for a serious disease of citrus known as huanglongbing (citrus greening disease). USDA-ARS researchers recently established a program for screening citrus germplasm for resistance to the di...

The LOV protein of Xanthomonas citri subsp. citri plays a significant role in the counteraction of plant immune responses during citrus canker.

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Ivana Kraiselburd

Full Text Available Pathogens interaction with a host plant starts a set of immune responses that result in complex changes in gene expression and plant physiology. Light is an important modulator of plant defense response and recent studies have evidenced the novel influence of this environmental stimulus in the virulence of several bacterial pathogens. Xanthomonas citri subsp. citri is the bacterium responsible for citrus canker disease, which affects most citrus cultivars. The ability of this bacterium to colonize host plants is influenced by bacterial blue-light sensing through a LOV-domain protein and disease symptoms are considerably altered upon deletion of this protein. In this work we aimed to unravel the role of this photoreceptor during the bacterial counteraction of plant immune responses leading to citrus canker development. We performed a transcriptomic analysis in Citrus sinensis leaves inoculated with the wild type X. citri subsp. citri and with a mutant strain lacking the LOV protein by a cDNA microarray and evaluated the differentially regulated genes corresponding to specific biological processes. A down-regulation of photosynthesis-related genes (together with a corresponding decrease in photosynthesis rates was observed upon bacterial infection, this effect being more pronounced in plants infected with the lov-mutant bacterial strain. Infection with this strain was also accompanied with the up-regulation of several secondary metabolism- and defense response-related genes. Moreover, we found that relevant plant physiological alterations triggered by pathogen attack such as cell wall fortification and tissue disruption were amplified during the lov-mutant strain infection. These results suggest the participation of the LOV-domain protein from X. citri subsp. citri in the bacterial counteraction of host plant defense response, contributing in this way to disease development.

The LOV Protein of Xanthomonas citri subsp. citri Plays a Significant Role in the Counteraction of Plant Immune Responses during Citrus Canker

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Kraiselburd, Ivana; Daurelio, Lucas D.; Tondo, María Laura; Merelo, Paz; Cortadi, Adriana A.; Talón, Manuel; Tadeo, Francisco R.; Orellano, Elena G.

2013-01-01

Pathogens interaction with a host plant starts a set of immune responses that result in complex changes in gene expression and plant physiology. Light is an important modulator of plant defense response and recent studies have evidenced the novel influence of this environmental stimulus in the virulence of several bacterial pathogens. Xanthomonas citri subsp. citri is the bacterium responsible for citrus canker disease, which affects most citrus cultivars. The ability of this bacterium to colonize host plants is influenced by bacterial blue-light sensing through a LOV-domain protein and disease symptoms are considerably altered upon deletion of this protein. In this work we aimed to unravel the role of this photoreceptor during the bacterial counteraction of plant immune responses leading to citrus canker development. We performed a transcriptomic analysis in Citrus sinensis leaves inoculated with the wild type X. citri subsp. citri and with a mutant strain lacking the LOV protein by a cDNA microarray and evaluated the differentially regulated genes corresponding to specific biological processes. A down-regulation of photosynthesis-related genes (together with a corresponding decrease in photosynthesis rates) was observed upon bacterial infection, this effect being more pronounced in plants infected with the lov-mutant bacterial strain. Infection with this strain was also accompanied with the up-regulation of several secondary metabolism- and defense response-related genes. Moreover, we found that relevant plant physiological alterations triggered by pathogen attack such as cell wall fortification and tissue disruption were amplified during the lov-mutant strain infection. These results suggest the participation of the LOV-domain protein from X. citri subsp. citri in the bacterial counteraction of host plant defense response, contributing in this way to disease development. PMID:24260514

Effects of soil-applied imidacloprid on Asian citrus psyllid (Hemiptera: Psyllidae) feeding behavior

Science.gov (United States)

The Asian citrus psyllid, Diaphorina citri Kuwayama is one of the most important pests of citrus due to its status as a vector of Candidatus Liberibacter asiaticus, the bacterium associated with huanglongbing (HLB) disease. The use of insecticides for vector control is the primary method of managing...

Whole plant destructive screening for huanglongbing susceptibility with conetainer seedlings exposed to no-choice Asian citrus psyllid inoculation

Science.gov (United States)

Huanglongbing (HLB) is associated with Candidatus Liberibacter asiaticus (Las) and is vectored by the Asian Citrus Psyllid (ACP, Diaphorina citri). HLB is devastating the Florida citrus industry, with production reduced by 60 percent in the last 12 years, and HLB is considered the greatest threat to...

The quest for a non-vector psyllid: Natural variation in acquisition and transmission of the huanglongbing pathogen 'Candidatus Liberibacter asiaticus' by Asian citrus psyllid isofemale lines

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Genetic variability in insect vectors is valuable to study vector competence determinants and to select non-vector populations that may help reduce the spread of vector-borne pathogens. We collected and tested vector competency of 15 isofemale lines of Asian citrus psyllid (ACP) Diaphorina citri, v...

Incidence of 'Candidatus Liberibacter asiaticus' in a Florida population of Asian citrus psyllid

Science.gov (United States)

A study was conducted to assess the incidence of a bacterium ‘Candidatus Liberibacter asiaticus’ in a Florida population of Asian citrus psyllid (ACP), Diaphorina citri. The bacterium is the presumed causal agent of Asiatic huanglongbing, a serious citrus disease also known as citrus greening or yel...

Phagostimulants for the Asian citrus psyllid also elicit volatile release from citrus leaves

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Chemical cues that elicit orientation by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), are of great interest because it is the primary vector of the causal pathogen of citrus greening disease. We identified an optimal blend ratio of formic and acetic acids that stimulate...

A phagostimulant blend for the Asian citrus psyllid

Science.gov (United States)

Chemical cues that condition orientation by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), are of great interest because it is the primary vector of the causal pathogen of citrus greening disease. Previous work in our lab identified a blend of formic and acetic acids as s...

Evidence of behavior-based utilization by the Asian citrus psyllid of a combination of UV and green or yellow wavelengths.

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Thomson M Paris

Full Text Available The Asian citrus psyllid, Diaphorina citri, vectors huanglongbing (HLB, the most serious disease affecting citrus globally. D. citri and HLB have spread to the major citrus growing regions of North America causing billions of dollars of damage in Florida alone. The visual behavior of D. citri is not well characterized and more knowledge is needed to improve attractive traps for monitoring and control of the D. citri. Bioassays were conducted to evaluate attraction to light transmitted through different colored filters. The addition of ultra-violet light (< 400 nm enhanced attraction of D. citri to transparent visual targets made of green or yellow filters. However, attraction to blue targets was unaffected by UV light. This is the first study to demonstrate a phytophagous insect responding to a hue that is a combination of long and short wavelengths. Further testing is needed to determine how D. citri uses such discriminatory powers in the field. Our results further imply that D. citri utilize color vision, as the less intense yellow and green hues were chosen over white light. In summary, this research provides an increased understanding of D. citri visual behavior and can be used for the development of a more attractive D. citri trap than those currently available.

Insights into xanthomonas axonopodis pv. Citri biofilm through proteomics

KAUST Repository

Zimaro, Tamara

2013-08-07

Background: Xanthomonas axonopodis pv. Citri (X. a. pv. Citri) causes citrus canker that can result in defoliation and premature fruit drop with significant production losses worldwide. Biofilm formation is an important process in bacterial pathogens and several lines of evidence suggest that in X. a. pv. Citri this process is a requirement to achieve maximal virulence since it has a major role in host interactions. In this study, proteomics was used to gain further insights into the functions of biofilms. Results: In order to identify differentially expressed proteins, a comparative proteomic study using 2D difference gel electrophoresis was carried out on X. a. pv. Citri mature biofilm and planktonic cells. The biofilm proteome showed major variations in the composition of outer membrane proteins and receptor or transport proteins. Among them, several porins and TonB-dependent receptor were differentially regulated in the biofilm compared to the planktonic cells, indicating that these proteins may serve in maintaining specific membrane-associated functions including signaling and cellular homeostasis. In biofilms, UDP-glucose dehydrogenase with a major role in exopolysaccharide production and the non-fimbrial adhesin YapH involved in adherence were over-expressed, while a polynucleotide phosphorylase that was demonstrated to negatively control biofilm formation in E. coli was down-regulated. In addition, several proteins involved in protein synthesis, folding and stabilization were up-regulated in biofilms. Interestingly, some proteins related to energy production, such as ATP-synthase were down-regulated in biofilms. Moreover, a number of enzymes of the tricarboxylic acid cycle were differentially expressed. In addition, X. a. pv. Citri biofilms also showed down-regulation of several antioxidant enzymes. The respective gene expression patterns of several identified proteins in both X. a. pv. Citri mature biofilm and planktonic cells were evaluated by

Reaction of detached leaves of different varieties of sweet orange (Citrus sinensis L. Osbeck) to inoculation with Xanthomonas citri subsp. citri (ex Hasse) Gabriel et al.

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Asiatic citrus canker (ACC) (caused by Xanthomonas citri subsp citri, Xcc) is a major disease of citrus in wet tropical and subtropical production regions. Screening for resistance is important to breeding programs. The objective of this study was to evaluate a detached leaf method to compare nine d...

Thresholds for HLB vector control in infected commercial citrus and compatibility with biological control

OpenAIRE

Monzo, C.; Hendricks, K.; Roberts, P.; Stansly, P. A.

2014-01-01

Control of the HLB vector, Diaphorina citri Kuwayama, is considered a basic component for management this disease, even in a high HLB incidence scenario. Such control is mostly chemically oriented. However, over use of insecticides would increase costs and be incompatible with biological control. Establishment of economic thresholds for psyllid control under different price scenarios could optimize returns on investment.

Metabolic Interplay between the Asian Citrus Psyllid and Its Profftella Symbiont: An Achilles' Heel of the Citrus Greening Insect Vector.

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John S Ramsey

Full Text Available 'Candidatus Liberibacter asiaticus' (CLas, the bacterial pathogen associated with citrus greening disease, is transmitted by Diaphorina citri, the Asian citrus psyllid. Interactions among D. citri and its microbial endosymbionts, including 'Candidatus Profftella armatura', are likely to impact transmission of CLas. We used quantitative mass spectrometry to compare the proteomes of CLas(+ and CLas(- populations of D. citri, and found that proteins involved in polyketide biosynthesis by the endosymbiont Profftella were up-regulated in CLas(+ insects. Mass spectrometry analysis of the Profftella polyketide diaphorin in D. citri metabolite extracts revealed the presence of a novel diaphorin-related polyketide and the ratio of these two polyketides was changed in CLas(+ insects. Insect proteins differentially expressed between CLas(+ and CLas(- D. citri included defense and immunity proteins, proteins involved in energy storage and utilization, and proteins involved in endocytosis, cellular adhesion, and cytoskeletal remodeling which are associated with microbial invasion of host cells. Insight into the metabolic interdependence between the insect vector, its endosymbionts, and the citrus greening pathogen reveals novel opportunities for control of this disease, which is currently having a devastating impact on citrus production worldwide.

POTENTIAL DISTRIBUTION OF ORANGE JASMINE (Murraya paniculata IN MEXICO

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José López-Collado

2013-04-01

Full Text Available Orange jasmine (OJ is a common ornamental plant used as green hedge in public and private gardens in Mexico. It also hosts Huanglongbing, a worldwide citrus disease and its vector, Diaphorina citri. For risk analysis and management purpose is important to know its geographic distribution. The potential distribution of OJ was calculated in Mexico using a deductive approach. Based on temperature and precipitation requirements, a relative suitability index was computed by combining the normalized values of both variables. The distribution was overlapped with captures of D. citri to check their spatial similarity. The results showed that the potential of occurrence is high in the Pacific and Gulf of México coastal states, including the Yucatán peninsula, and the lowest values appeared in the north-western states. The OJ distribution overlaps with Huanglongbing occurrence and coincided with captures of D. citri for most of the suitable area but D. citri captures extended beyond the optimal OJ distribution values in the northern regions of México.

Identification of Immunity-Related Genes in Dialeurodes citri against Entomopathogenic Fungus Lecanicillium attenuatum by RNA-Seq Analysis.

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Shijiang Yu

Full Text Available Dialeurodes citri is a major pest in citrus producing areas, and large-scale outbreaks have occurred increasingly often in recent years. Lecanicillium attenuatum is an important entomopathogenic fungus that can parasitize and kill D. citri. We separated the fungus from corpses of D. citri larvae. However, the sound immune defense system of pests makes infection by an entomopathogenic fungus difficult. Here we used RNA sequencing technology (RNA-Seq to build a transcriptome database for D. citri and performed digital gene expression profiling to screen genes that act in the immune defense of D. citri larvae infected with a pathogenic fungus. De novo assembly generated 84,733 unigenes with mean length of 772 nt. All unigenes were searched against GO, Nr, Swiss-Prot, COG, and KEGG databases and a total of 28,190 (33.3% unigenes were annotated. We identified 129 immunity-related unigenes in transcriptome database that were related to pattern recognition receptors, information transduction factors and response factors. From the digital gene expression profile, we identified 441 unigenes that were differentially expressed in D. citri infected with L. attenuatum. Through calculated Log2Ratio values, we identified genes for which fold changes in expression were obvious, including cuticle protein, vitellogenin, cathepsin, prophenoloxidase, clip-domain serine protease, lysozyme, and others. Subsequent quantitative real-time polymerase chain reaction analysis verified the results. The identified genes may serve as target genes for microbial control of D. citri.

Identification of Immunity-Related Genes in Dialeurodes citri against Entomopathogenic Fungus Lecanicillium attenuatum by RNA-Seq Analysis.

Science.gov (United States)

Yu, Shijiang; Ding, Lili; Luo, Ren; Li, Xiaojiao; Yang, Juan; Liu, Haoqiang; Cong, Lin; Ran, Chun

2016-01-01

Dialeurodes citri is a major pest in citrus producing areas, and large-scale outbreaks have occurred increasingly often in recent years. Lecanicillium attenuatum is an important entomopathogenic fungus that can parasitize and kill D. citri. We separated the fungus from corpses of D. citri larvae. However, the sound immune defense system of pests makes infection by an entomopathogenic fungus difficult. Here we used RNA sequencing technology (RNA-Seq) to build a transcriptome database for D. citri and performed digital gene expression profiling to screen genes that act in the immune defense of D. citri larvae infected with a pathogenic fungus. De novo assembly generated 84,733 unigenes with mean length of 772 nt. All unigenes were searched against GO, Nr, Swiss-Prot, COG, and KEGG databases and a total of 28,190 (33.3%) unigenes were annotated. We identified 129 immunity-related unigenes in transcriptome database that were related to pattern recognition receptors, information transduction factors and response factors. From the digital gene expression profile, we identified 441 unigenes that were differentially expressed in D. citri infected with L. attenuatum. Through calculated Log2Ratio values, we identified genes for which fold changes in expression were obvious, including cuticle protein, vitellogenin, cathepsin, prophenoloxidase, clip-domain serine protease, lysozyme, and others. Subsequent quantitative real-time polymerase chain reaction analysis verified the results. The identified genes may serve as target genes for microbial control of D. citri.

New genes of Xanthomonas citri subsp. citri involved in pathogenesis and adaptation revealed by a transposon-based mutant library.

Science.gov (United States)

Laia, Marcelo L; Moreira, Leandro M; Dezajacomo, Juliana; Brigati, Joice B; Ferreira, Cristiano B; Ferro, Maria I T; Silva, Ana C R; Ferro, Jesus A; Oliveira, Julio C F

2009-01-16

Citrus canker is a disease caused by the phytopathogens Xanthomonas citri subsp. citri, Xanthomonas fuscans subsp. aurantifolli and Xanthomonas alfalfae subsp. citrumelonis. The first of the three species, which causes citrus bacterial canker type A, is the most widely spread and severe, attacking all citrus species. In Brazil, this species is the most important, being found in practically all areas where citrus canker has been detected. Like most phytobacterioses, there is no efficient way to control citrus canker. Considering the importance of the disease worldwide, investigation is needed to accurately detect which genes are related to the pathogen-host adaptation process and which are associated with pathogenesis. Through transposon insertion mutagenesis, 10,000 mutants of Xanthomonas citri subsp. citri strain 306 (Xcc) were obtained, and 3,300 were inoculated in Rangpur lime (Citrus limonia) leaves. Their ability to cause citrus canker was analyzed every 3 days until 21 days after inoculation; a set of 44 mutants showed altered virulence, with 8 presenting a complete loss of causing citrus canker symptoms. Sequencing of the insertion site in all 44 mutants revealed that 35 different ORFs were hit, since some ORFs were hit in more than one mutant, with mutants for the same ORF presenting the same phenotype. An analysis of these ORFs showed that some encoded genes were previously known as related to pathogenicity in phytobacteria and, more interestingly, revealed new genes never implicated with Xanthomonas pathogenicity before, including hypothetical ORFs. Among the 8 mutants with no canker symptoms are the hrpB4 and hrpX genes, two genes that belong to type III secretion system (TTSS), two hypothetical ORFS and, surprisingly, the htrA gene, a gene reported as involved with the virulence process in animal-pathogenic bacteria but not described as involved in phytobacteria virulence. Nucleic acid hybridization using labeled cDNA probes showed that some of the

A plant pathogenic bacterium exploits the tricarboxylic acid cycle metabolic pathway of its insect vector

Science.gov (United States)

Nehela, Yasser; Hijaz, Faraj; Vincent, Christopher I.

2018-01-01

ABSTRACT Huanglongbing in citrus is caused by a phloem-limited, uncultivable, gram-negative α-proteobacterium, Candidatus Liberibacter asiaticus (CLas). CLas is transmitted by the phloem-sucking insect, Diaphorina citri (Hemiptera: Liviidae), in a persistent, circulative, and propagative manner. In this study, we investigated the metabolomic and respiration rates changes in D. citri upon infection with CLas using gas chromatography-mass spectrometry (GC-MS) and gas exchange analysis. The level of glycine, L-serine, L-threonine, and gamma-amino butyric acid were higher in CLas-infected D. citri, while L-proline, L-aspartic acid, and L-pyroglutamic acid were lower in CLas-infected D. citri compared with the control. Citric acid was increased in CLas-infected D. citri, whereas malic and succinic acids were reduced. Interestingly, most of the reduced metabolites such as malate, succinate, aspartate, and L-proline are required for the growth of CLas. The increase in citric acid, serine, and glycine indicated that CLas induced glycolysis and the tricarboxylic acid cycle (TCA) in its vector. In agreement with the GC-MS results, the gene expression results also indicated that glycolysis and TCA were induced in CLas-infected D. citri and this was accompanied with an increases in respiration rate. Phosphoric acid and most of the sugar alcohols were higher in CLas-infected D. citri, indicating a response to the biotic stress or cell damage. Only slight increases in the levels of few sugars were observed in CLas-infected D. citri, which indicated that sugars are tightly regulated by D. citri. Our results indicated that CLas induces nutrient and energetic stress in its host insect. This study may provide some insights into the mechanism of colonization of CLas in its vector. PMID:28594267

Ectopic accumulation of linalool confers resistance to Xanthomonas citri subsp. citri in transgenic sweet orange plants.

Science.gov (United States)

Shimada, Takehiko; Endo, Tomoko; Rodríguez, Ana; Fujii, Hiroshi; Goto, Shingo; Matsuura, Takakazu; Hojo, Yuko; Ikeda, Yoko; Mori, Izumi C; Fujikawa, Takashi; Peña, Leandro; Omura, Mitsuo

2017-05-01

In order to clarify whether high linalool content in citrus leaves alone induces strong field resistance to citrus canker caused by Xanthomonas citri subsp. citri (Xcc), and to assess whether this trait can be transferred to a citrus type highly sensitive to the bacterium, transgenic 'Hamlin' sweet orange (Citrus sinensis L. Osbeck) plants over-expressing a linalool synthase gene (CuSTS3-1) were generated. Transgenic lines (LIL) with the highest linalool content showed strong resistance to citrus canker when spray inoculated with the bacterium. In LIL plants inoculated by wounding (multiple-needle inoculation), the linalool level was correlated with the repression of the bacterial titer and up-regulation of defense-related genes. The exogenous application of salicylic acid, methyl jasmonate or linalool triggered responses similar to those constitutively induced in LIL plants. The linalool content in Ponkan mandarin leaves was significantly higher than that of leaves from six other representative citrus genotypes with different susceptibilities to Xcc. We propose that linalool-mediated resistance might be unique to citrus tissues accumulating large amounts of volatile organic compounds in oil cells. Linalool might act not only as a direct antibacterial agent, but also as a signal molecule involved in triggering a non-host resistance response against Xcc. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Structural and physiological analyses of the alkanesulphonate-binding protein (SsuA of the citrus pathogen Xanthomonas citri.

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Fabiano Tófoli de Araújo

Full Text Available BACKGROUND: The uptake of sulphur-containing compounds plays a pivotal role in the physiology of bacteria that live in aerobic soils where organosulfur compounds such as sulphonates and sulphate esters represent more than 95% of the available sulphur. Until now, no information has been available on the uptake of sulphonates by bacterial plant pathogens, particularly those of the Xanthomonas genus, which encompasses several pathogenic species. In the present study, we characterised the alkanesulphonate uptake system (Ssu of Xanthomonas axonopodis pv. citri 306 strain (X. citri, the etiological agent of citrus canker. METHODOLOGY/PRINCIPAL FINDINGS: A single operon-like gene cluster (ssuEDACB that encodes both the sulphur uptake system and enzymes involved in desulphurisation was detected in the genomes of X. citri and of the closely related species. We characterised X. citri SsuA protein, a periplasmic alkanesulphonate-binding protein that, together with SsuC and SsuB, defines the alkanesulphonate uptake system. The crystal structure of SsuA bound to MOPS, MES and HEPES, which is herein described for the first time, provides evidence for the importance of a conserved dipole in sulphate group coordination, identifies specific amino acids interacting with the sulphate group and shows the presence of a rather large binding pocket that explains the rather wide range of molecules recognised by the protein. Isolation of an isogenic ssuA-knockout derivative of the X. citri 306 strain showed that disruption of alkanesulphonate uptake affects both xanthan gum production and generation of canker lesions in sweet orange leaves. CONCLUSIONS/SIGNIFICANCE: The present study unravels unique structural and functional features of the X. citri SsuA protein and provides the first experimental evidence that an ABC uptake system affects the virulence of this phytopathogen.

The molybdate-binding protein (ModA) of the plant pathogen Xanthomonas axonopodis pv. citri.

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Balan, Andrea; Santacruz, Carolina P; Moutran, Alexandre; Ferreira, Rita C C; Medrano, Francisco J; Pérez, Carlos A; Ramos, Carlos H I; Ferreira, Luís C S

2006-12-01

The modABC operon of phytopathogen Xanthomonas axonopodis pv. citri (X. citri) encodes a putative ABC transporter involved in the uptake of the molybdate and tungstate anions. Sequence analyses showed high similarity values of ModA orthologs found in X. campestris pv. campestris (X. campestris) and Escherichia coli. The X. citri modA gene was cloned in pET28a and the recombinant protein, expressed in the E. coli BL21 (DE3) strain, purified by immobilized metal affinity chromatography. The purified protein remained soluble and specifically bound molybdate and tungstate with K(d) 0.29+/-0.12 microM and 0.58+/-0.14 microM, respectively. Additionally binding of molybdate drastically enhanced the thermal stability of the recombinant ModA as compared to the apoprotein. This is the first characterization of a ModA ortholog expressed by a phytopathogen and represents an important tool for functional, biochemical and structural analyses of molybdate transport in Xanthomonas species.

Xanthomonas citri ssp. citri requires the outer membrane porin OprB for maximal virulence and biofilm formation.

Science.gov (United States)

Ficarra, Florencia A; Grandellis, Carolina; Galván, Estela M; Ielpi, Luis; Feil, Regina; Lunn, John E; Gottig, Natalia; Ottado, Jorgelina

2017-06-01

Xanthomonas citri ssp. citri (Xcc) causes canker disease in citrus, and biofilm formation is critical for the disease cycle. OprB (Outer membrane protein B) has been shown previously to be more abundant in Xcc biofilms compared with the planktonic state. In this work, we showed that the loss of OprB in an oprB mutant abolishes bacterial biofilm formation and adherence to the host, and also compromises virulence and efficient epiphytic survival of the bacteria. Moreover, the oprB mutant is impaired in bacterial stress resistance. OprB belongs to a family of carbohydrate transport proteins, and the uptake of glucose is decreased in the mutant strain, indicating that OprB transports glucose. Loss of OprB leads to increased production of xanthan exopolysaccharide, and the carbohydrate intermediates of xanthan biosynthesis are also elevated in the mutant. The xanthan produced by the mutant has a higher viscosity and, unlike wild-type xanthan, completely lacks pyruvylation. Overall, these results suggest that Xcc reprogrammes its carbon metabolism when it senses a shortage of glucose input. The participation of OprB in the process of biofilm formation and virulence, as well as in metabolic changes to redirect the carbon flux, is discussed. Our results demonstrate the importance of environmental nutrient supply and glucose uptake via OprB for Xcc virulence. © 2016 BSPP AND JOHN WILEY & SONS LTD.

Quality Matters: Influences of Citrus Flush Physicochemical Characteristics on Population Dynamics of the Asian Citrus Psyllid (Hemiptera: Liviidae.

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Mamoudou Sétamou

Full Text Available Studies were conducted to relate the influence of the physical characteristics, leaf nutrient content and phloem sap amino acid concentration of citrus flush shoots on the densities of various Diaphorina citri life stages. Adult D. citri preferentially selected young shoots for feeding and numbers of D. citri immatures were positively correlated with flush shoot softness. Young flush shoots had higher concentrations of macro and micro nutrients relative to mature ones and this was associated with higher densities of all D. citri life stages. All D. citri life stages were positively correlated with higher nitrogen-carbon (N:C, nitrogen:sulfur (N:S and nitrogen:calcium (N:Ca ratios in leaf tissue, while densities of adults were negatively related to calcium, manganese and boron levels. Concentrations of total and essential amino acids were highest in phloem sap of young expanding flush shoots in both grapefruit and lemon, but dramatically declined as flush shoots matured. The sulfur-containing amino acids cystine, methionine and taurine occurred only in younger flush shoots. In contrast, cystathionine was only present in phloem sap of mature shoots. These results clearly indicate that young citrus flush shoots are a nutritionally richer diet relative to mature shoots, thus explaining their preference by D. citri for feeding and reproduction. Conversely, tissue hardness and the lower nutritional quality of mature flush shoots may limit oviposition and immature development. The data suggest that both physical characteristics and nutritional composition of flush shoots and their phloem sap are important factors regulating host colonization and behavior of D. citri, and this interaction can impact the dynamics and spread of HLB in citrus groves.

Quality Matters: Influences of Citrus Flush Physicochemical Characteristics on Population Dynamics of the Asian Citrus Psyllid (Hemiptera: Liviidae)

Science.gov (United States)

Simpson, Catherine R.; Alabi, Olufemi J.; Nelson, Shad D.; Telagamsetty, Srilakshmi; Jifon, John L.

2016-01-01

Studies were conducted to relate the influence of the physical characteristics, leaf nutrient content and phloem sap amino acid concentration of citrus flush shoots on the densities of various Diaphorina citri life stages. Adult D. citri preferentially selected young shoots for feeding and numbers of D. citri immatures were positively correlated with flush shoot softness. Young flush shoots had higher concentrations of macro and micro nutrients relative to mature ones and this was associated with higher densities of all D. citri life stages. All D. citri life stages were positively correlated with higher nitrogen-carbon (N:C), nitrogen:sulfur (N:S) and nitrogen:calcium (N:Ca) ratios in leaf tissue, while densities of adults were negatively related to calcium, manganese and boron levels. Concentrations of total and essential amino acids were highest in phloem sap of young expanding flush shoots in both grapefruit and lemon, but dramatically declined as flush shoots matured. The sulfur-containing amino acids cystine, methionine and taurine occurred only in younger flush shoots. In contrast, cystathionine was only present in phloem sap of mature shoots. These results clearly indicate that young citrus flush shoots are a nutritionally richer diet relative to mature shoots, thus explaining their preference by D. citri for feeding and reproduction. Conversely, tissue hardness and the lower nutritional quality of mature flush shoots may limit oviposition and immature development. The data suggest that both physical characteristics and nutritional composition of flush shoots and their phloem sap are important factors regulating host colonization and behavior of D. citri, and this interaction can impact the dynamics and spread of HLB in citrus groves. PMID:28030637

Acción inhibitoria de una cepa de Zymomonas mobilis mobilis aislada de caña de azúcar sobre Xanthomonas citri subsp. citri, agente causal de la cancrosis de los cítricos Inhibition of Xanthomonas citri subsp. citri, causal agent of citrus canker, by a strain of Zymomonas mobilis mobilis isolated from sugarcane

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María E. Romero

2008-06-01

Full Text Available Zymomonas mobilis mobilis (Zm produce factores antimicrobianos que actúan sobre un amplio espectro de microorganismos patógenos para el hombre, animales y plantas. Un problema importante a resolver en los tratamientos con antimicrobianos, es el desarrollo de resistencia a compuestos empleados actualmente, no siendo las bacterias fitopatógenas una excepción. En el presente trabajo se realizaron ensayos de antagonismo con células (pruebas de estrías cruzadas y sobrenadantes concentrados (Sc (por difusión en agar preparados a partir de cultivos de Zm (aislada de jugo de caña de azúcar producido en Tucumán, frente a la bacteria causal de la cancrosis: Xanthomonas citri subsp. citri. Se evaluaron aislamientos de Xcc sensibles (Xc y resistentes (Xcr a compuestos a base de cobre. Los resultados obtenidos mostraron que la bacteria testigo fue inhibida totalmente por las células de Zm, ejerciendo un efecto bactericida. En los ensayos de difusión en el agar se observó que tanto Xc, como Xcr fueron sensibles al Sc de Zm. Se sabe, por estudios anteriores, que los metabolitos de Zymomonas tienen un efecto deletéreo en la membrana celular de E. coli AB1133, inhibiéndose la respiración de la bacteria inmediatamente de agregado Sc (60 UA. En el presente trabajo se observó el mismo efecto, inhibición total de la respiración en Xc, luego del agregado del Sc (60 UA. Por lo observado, se deduce que el blanco de acción de los metabolitos antimicrobianos de Sc en Xc, sería el mismo que el de E. coli AB1133. Con los resultados obtenidos se considera de interés encarar el estudio de los compuestos de Zm para ser empleados en el control de enfermedades que afectan los cultivos de valor económico de la región, como es el caso de la cancrosis, como así también profundizar acerca de la acción de dichos metabolitos en la membrana de Xanthomonas citri subsp. citri.Zymomonas mobilis mobilis (Zm produces antimicrobial factors, which have an effect on

Evidence that ‘flying dragon’ trifoliate orange delays HLB symptom expression for four sweet orange cultivars, Tahiti lime and Okitsu mandarin

OpenAIRE

Stuchi, E. S.; Reiff, E. T.; Sempionato, O. R.; Parolin, L. G.; Toledo, D. A.

2014-01-01

Huanglongbing (HLB), caused by Candidatus Liberibacter asiaticus and vectored by Diaphorina citri, was first reported in 2004 in Brazil and is currently widespread in São Paulo State. Brazil is the world’s largest sweet orange producer and has 49,000 ha cultivated with ‘Tahiti’ lime acid lime. Mandarin cultivation represents 5.5% of total citrus production in the country. In 2001, three experiments were planted in the Citrus Experimental Station (EECB), Bebedouro, Northern São Paulo State, wh...

In vitro control of Alternaria citri using antifungal potentials of ...

African Journals Online (AJOL)

In vitro control of Alternaria citri using antifungal potentials of Trichoderma species. Asma Murtaza, Shazia Shafique, Tehmina Anjum, Sobiya Shafique. Abstract. The antifungal potential of five species of Trichoderma viz., Trichoderma viride, Trichoderma aureoviride, Trichoderma reesei, Trichoderma koningii and ...

Geïntegreerde bestrijding van citruswolluis Planococcus citri in roos

NARCIS (Netherlands)

Pijnakker, J.; Leman, A.; Hennekam, M.

2013-01-01

Citruswolluis, Planococcus citri (Risso), is een van de belangrijkste plagen geworden in de rozenteelt onder glas. Er is inmiddels veel praktijkervaring met de toepassing van biologische bestrijders om haarden uit te roeien. Voor de professionele tuinbouw is de effectiviteit van deze aanpak

First report in Ghana of #Xanthomonas citri# pv. #mangiferaeindicae# causing mango bacterial canker on #Mangifera indica#

OpenAIRE

Pruvost, Olivier; Boyer, Claudine; Vital, Karine; Gagnevin, Lionel; De Bruno Austin, L.; Rey, Jean-Yves

2011-01-01

Bacterial canker of mango (or bacterial black spot), caused by Xanthomonas citri pv. mangiferaeindicae, is an economically important disease in tropical and subtropical producing areas (1). X. citri pv. mangiferaeindicae can cause severe infection in a wide range of mango cultivars and induces raised, angular, black leaf lesions, sometimes with a chlorotic halo. Several months after infection, leaf lesions dry and turn light brown or ash gray. Severe leaf infection may result in abscission. F...

Molecular Effects of Irradiation (Cobalt-60 on the Control of Panonychus citri (Acari: Tetranychidae

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Ke Zhang

2015-11-01

Full Text Available The effective dose of irradiation to control pest mites in quarantine has been studied extensively, but the molecular mechanisms underlying the effects of the irradiation on mites are largely unknown. In this study, exposure to 400 Gy of γ rays had significant (p < 0.05 effects on the adult survival, fecundity and egg viability of Panonychus citri. The irradiation caused the degradation of the DNA of P. citri adults and damaged the plasma membrane system of the egg, which led to condensed nucleoli and gathered yolk. Additionally, the transcriptomes and gene expression profiles between irradiated and non-irradiated mites were compared, and three digital gene expression libraries were assembled and analyzed. The differentially expressed genes were putatively involved in apoptosis, cell death and the cell cycle. Finally, the expression profiles of some related genes were studied using quantitative real-time PCR. Our study provides valuable information on the changes in the transcriptome of irradiated P. citri, which will facilitate a better understanding of the molecular mechanisms that cause the sterility induced by irradiation.

A Plant Bacterial Pathogen Manipulates Its Insect Vector's Energy Metabolism

Science.gov (United States)

Hijaz, Faraj; Ebert, Timothy A.; Rogers, Michael E.

2016-01-01

ABSTRACT Insect-transmitted plant-pathogenic bacteria may alter their vectors' fitness, survival, behavior, and metabolism. Because these pathogens interact with their vectors on the cellular and organismal levels, potential changes at the biochemical level might occur. “Candidatus Liberibacter asiaticus” (CLas) is transmitted in a persistent, circulative, and propagative manner. The genome of CLas revealed the presence of an ATP translocase that mediates the uptake of ATP and other nucleotides from medium to achieve its biological processes, such as growth and multiplication. Here, we showed that the levels of ATP and many other nucleotides were significantly higher in CLas-infected than healthy psyllids. Gene expression analysis showed upregulation for ATP synthase subunits, while ATPase enzyme activity showed a decrease in ATPase activity. These results indicated that CLas stimulated Diaphorina citri to produce more ATP and many other energetic nucleotides, while it may inhibit their consumption by the insect. As a result of ATP accumulation, the adenylated energy charge (AEC) increased and the AMP/ATP and ADP/ATP ratios decreased in CLas-infected D. citri psyllids. Survival analysis confirmed a shorter life span for CLas-infected D. citri psyllids. In addition, electropenetrography showed a significant reduction in total nonprobing time, salivation time, and time from the last E2 (phloem ingestion) to the end of recording, indicating that CLas-infected psyllids were at a higher hunger level and they tended to forage more often. This increased feeding activity reflects the CLas-induced energetic stress. In conclusion, CLas alters the energy metabolism of its psyllid vector, D. citri, in order to secure its need for energetic nucleotides. IMPORTANCE Insect transmission of plant-pathogenic bacteria involves propagation and circulation of the bacteria within their vectors. The transmission process is complex and requires specific interactions at the molecular

[Identification of two varieties of Citri Fructus by fingerprint and chemometrics].

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Su, Jing-hua; Zhang, Chao; Sun, Lei; Gu, Bing-ren; Ma, Shuang-cheng

2015-06-01

Citri Fructus identification by fingerprint and chemometrics was investigated in this paper. Twenty-three Citri Fructus samples were collected which referred to two varieties as Cirtus wilsonii and C. medica recorded in Chinese Pharmacopoeia. HPLC chromatograms were obtained. The components were partly identified by reference substances, and then common pattern was established for chemometrics analysis. Similarity analysis, principal component analysis (PCA) , partial least squares-discriminant analysis (PLS-DA) and hierarchical cluster analysis heatmap were applied. The results indicated that C. wilsonii and C. medica could be ideally classified with common pattern contained twenty-five characteristic peaks. Besides, preliminary pattern recognition had verified the chemometrics analytical results. Absolute peak area (APA) was used for relevant quantitative analysis, results showed the differences between two varieties and it was valuable for further quality control as selection of characteristic components.

Compuestos volátiles de brotes de limón persa y limón mexicano asociados a síntomas del HLB (Huanglongbing).

OpenAIRE

Mendoza Peña, Estrella

2014-01-01

El Huanglongbing (HLB) es una enfermedad de los cítricos que se caracteriza por causar áreas moteadas en hojas, generar frutos deformes y decolorados, semillas abortivas, causar pérdida paulatina de la producción y provocar que la muerte de los árboles infectados sea inevitable. El agente asociado a esta enfermedad es la bacteria Candidatus Liberibacter ssp., y en México es dispersada por el vector Diaphorina citri Kuwayama (Hemiptera: Liviidae). En el presente estudio, mediante el método de ...

A model for Huanglongbing spread between citrus plants including delay times and human intervention

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Vilamiu, Raphael G. d'A.; Ternes, Sonia; Braga, Guilherme A.; Laranjeira, Francisco F.

2012-09-01

The objective of this work was to present a compartmental deterministic mathematical model for representing the dynamics of HLB disease in a citrus orchard, including delay in the disease's incubation phase in the plants, and a delay period on the nymphal stage of Diaphorina citri, the most important HLB insect vector in Brazil. Numerical simulations were performed to assess the possible impacts of human detection efficiency of symptomatic plants, as well as the influence of a long incubation period of HLB in the plant.

Application of droplet digital PCR for quantitative detection of Spiroplasma citri in comparison with real time PCR.

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Yogita Maheshwari

Full Text Available Droplet digital polymerase chain reaction (ddPCR is a method for performing digital PCR that is based on water-oil emulsion droplet technology. It is a unique approach to measure the absolute copy number of nucleic acid targets without the need of external standards. This study evaluated the applicability of ddPCR as a quantitative detection tool for the Spiroplasma citri, causal agent of citrus stubborn disease (CSD in citrus. Two sets of primers, SP1, based on the spiral in housekeeping gene, and a multicopy prophage gene, SpV1 ORF1, were used to evaluate ddPCR in comparison with real time (quantitative PCR (qPCR for S. citri detection in citrus tissues. Standard curve analyses on tenfold dilution series showed that both ddPCR and qPCR exhibited good linearity and efficiency. However, ddPCR had a tenfold greater sensitivity than qPCR and accurately quantified up to one copy of spiralin gene. Receiver operating characteristic analysis indicated that the ddPCR methodology was more robust for diagnosis of CSD and the area under the curve was significantly broader compared to qPCR. Field samples were used to validate ddPCR efficacy and demonstrated that it was equal or better than qPCR to detect S. citri infection in fruit columella due to a higher pathogen titer. The ddPCR assay detected both the S. citri spiralin and the SpV1 ORF1 targets quantitatively with high precision and accuracy compared to qPCR assay. The ddPCR was highly reproducible and repeatable for both the targets and showed higher resilience to PCR inhibitors in citrus tissue extract for the quantification of S. citri compare to qPCR.

Different transcriptional response to Xanthomonas citri subsp. citri between kumquat and sweet orange with contrasting canker tolerance.

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Xing-Zheng Fu

Full Text Available Citrus canker disease caused by Xanthomonas citri subsp. citri (Xcc is one of the most devastating biotic stresses affecting the citrus industry. Meiwa kumquat (Fortunella crassifolia is canker-resistant, while Newhall navel orange (Citrus sinensis Osbeck is canker-sensitive. To understand the molecular mechanisms underlying the differences in responses to Xcc, transcriptomic profiles of these two genotypes following Xcc attack were compared by using the Affymetrix citrus genome GeneChip. A total of 794 and 1324 differentially expressed genes (DEGs were identified as canker-responsive genes in Meiwa and Newhall, respectively. Of these, 230 genes were expressed in common between both genotypes, while 564 and 1094 genes were only significantly expressed in either Meiwa or Newhall. Gene ontology (GO annotation and Singular Enrichment Analysis (SEA of the DEGs showed that genes related to the cell wall and polysaccharide metabolism were induced for basic defense in both Meiwa and Newhall, such as chitinase, glucanase and thaumatin-like protein. Moreover, apart from inducing basic defense, Meiwa showed specially upregulated expression of several genes involved in the response to biotic stimulus, defense response, and cation binding as comparing with Newhall. And in Newhall, abundant photosynthesis-related genes were significantly down-regulated, which may be in order to ensure the basic defense. This study revealed different molecular responses to canker disease in Meiwa and Newhall, affording insight into the response to canker and providing valuable information for the identification of potential genes for engineering canker tolerance in the future.

Oral delivery of double-stranded RNAs and siRNAs induces RNAi effects in the potato/tomato psyllid, Bactericerca cockerelli.

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Hada Wuriyanghan

Full Text Available The potato/tomato psyllid, Bactericerca cockerelli (B. cockerelli, and the Asian citrus psyllid, Diaphorina citri (D. citri, are very important plant pests, but they are also vectors of phloem-limited bacteria that are associated with two devastating plant diseases. B. cockerelli is the vector of Candidatus Liberibacter psyllaurous (solanacearum, which is associated with zebra chip disease of potatoes, and D. citri is the vector of Ca. Liberibacter asiaticus, which is associated with the Huanglongbing (citrus greening disease that currently threatens the entire Florida citrus industry. Here we used EST sequence information from D. citri to identify potential targets for RNA interference in B. cockerelli. We targeted ubiquitously expressed and gut-abundant mRNAs via injection and oral acquisition of double-stranded RNAs and siRNAs and were able to induce mortality in recipient psyllids. We also showed knockdown of target mRNAs, and that oral acquisition resulted primarily in mRNA knockdown in the psyllid gut. Concurrent with gene knockdown was the accumulation of target specific ∼ 21 nucleotide siRNAs for an abundant mRNA for BC-Actin. These results showed that RNAi can be a powerful tool for gene function studies in psyllids, and give support for continued efforts for investigating RNAi approaches as possible tools for psyllid and plant disease control.

Metabolomic comparative analysis of the phloem sap of curry leaf tree (Bergera koenegii), orange jasmine (Murraya paniculata), and Valencia sweet orange (Citrus sinensis) supports their differential responses to Huanglongbing.

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Killiny, Nabil

2016-11-01

Orange jasmine, Murraya paniculata and curry leaf tree, Bergera koenegii are alternative hosts for Diaphorina citri, the vector of Candidatus Liberibacter asiaticus (CLas), the pathogen of huanglongbing (HLB) in citrus. D. citri feeds on the phloem sap where CLas grows. It has been shown that orange jasmine was a better host than curry leaf tree to D. citri. In addition, CLas can infect orange jasmine but not curry leaf tree. Here, we compared the phloem sap composition of these 2 plants to the main host, Valencia sweet orange, Citrus sinensis. Phloem sap was analyzed by gas chromatography-mass spectrometry after trimethylsilyl derivatization. Orange jasmine was the highest in proteinogenic, non-proteinogenic amino acids, organic acids, as well as total metabolites. Valencia was the highest in mono- and disaccharides, and sugar alcohols. Curry leaf tree was the lowest in most of the metabolites as well as total metabolites. Interestingly, malic acid was high in Valencia and orange jasmine but was not detected in the curry leaf. On the other hand, tartaric acid which can prevent the formation of malic acid in Krebs cycle was high in curry leaf. The nutrient inadequacy of the phloem sap in curry leaf tree, especially the amino acids could be the reason behind the longer life cycle and the low survival of D. citri and the limitation of CLas growth on this host. Information obtained from this study may help in cultivation of CLas and development of artificial diet for rearing of D. citri.

Sexual transmission of a plant pathogenic bacterium, Candidatus Liberibacter asiaticus, between conspecific insect vectors during mating.

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Rajinder S Mann

Full Text Available Candidatus Liberibacter asiaticus is a fastidious, phloem-inhabiting, gram-negative bacterium transmitted by Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae. The bacterium is the presumed causal agent of huanglongbing (HLB, one of the most destructive and economically important diseases of citrus. We investigated whether Las is transmitted between infected and uninfected D. citri adults during courtship. Our results indicate that Las was sexually transmitted from Las-infected male D. citri to uninfected females at a low rate (<4% during mating. Sexual transmission was not observed following mating of infected females and uninfected males or among adult pairs of the same sex. Las was detected in genitalia of both sexes and also in eggs of infected females. A latent period of 7 days or more was required to detect the bacterium in recipient females. Rod shaped as well as spherical structures resembling Las were observed in ovaries of Las-infected females with transmission electron microscopy, but were absent in ovaries from uninfected D. citri females. The size of the rod shaped structures varied from 0.39 to 0.67 µm in length and 0.19 to 0.39 µm in width. The spherical structures measured from 0.61 to 0.80 µm in diameter. This investigation provides convincing evidence that a plant pathogenic bacterium is sexually transmitted from male to female insects during courtship and established evidence that bacteria persist in reproductive organs. Moreover, these findings provide an alternative sexually horizontal mechanism for the spread of Las within populations of D. citri, even in the absence of infected host trees.

Pengaruh Minyak Atsiri Asal Kulit Jeruk Manis dan Besar terhadap Perkembangan Tungau Panonychus citri (Acarina: Tetranychidae

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Mizu lstianto

2003-12-01

Full Text Available P. citri (Acarina: Tetranychidae is one of the economically important citrus pests in Indonesia. However, the association of this pest with its host is not well understood. The objective of this research was to evaluate the effects of essential oil extracted from sweet orange and pumello fruit peels on the development of P. citri. The research was conducted in the laboratory of IP2TP Tlekung Malang, using a complete random design consisted of six treatments. Fifteen mites were used for each treatment. The treatments were concentrations 10, 20, 40, and 80 ppm of essential oil. Parafin was used as a positive control because it was a diluting solution for the essential oil. The control received no application. The result showed that the main content of essential oil extracted from sweet orange and pumello was limonene. The essential oils prolonged the life cycle and reduced the fecundity of P. citri. The effects of essential oil extracted from pumello were found to be more pronounced than were from sweet orange. These differences might be due to the differences in the composition of the volatile compounds other than limonene.

Using mass spectrometry for identification of ABC transporters from Xanthomonas citri and mutants expressed in different growth conditions

International Nuclear Information System (INIS)

Faria, J.N.; Balan, A.; Paes Leme, A.F.

2012-01-01

Full text: Xanthomonas citri is a phytopathogenic bacterium that infects citrus plants causing significant losses for the economy. In our group, we have focused on the identification and characterization of ABC transport proteins of this bacterium, in order to determinate their function for growth in vitro and in vivo, during infection. ABC transporters represent one of the largest families of proteins, which transport since small molecules as ions up to oligopeptides and sugars. In prokaryotic cells many works have reported the ABC transport function in pathogenesis, resistance, biofilm formation, infectivity and DNA repair, but until our knowledge, there is no data related to these transporters and X. citri. So, In order to determinate which transporters are expressed in X. citri, we started a proteomic analysis based on mono and bi-dimensional gels associated to mass spectrometry analyses. After growing X. citri and two different mutants deleted for ssuA and nitA genes in LB and minimum media, cellular extracts were obtained and used for preparation of mono and bi-dimensional gels. Seven bands covering the expected mass of ABC transporter components (20 kDa to 50 kDa) in SDS-PAGE were cut off the gel, treated with trypsin and submitted to the MS for protein identification. The results of 2D gels were good enough and will serve as a standard for development of similar experiments in large scale. (author)

Structural and functional analyses of the putrescine binding protein PotF from Xanthomonas citri

International Nuclear Information System (INIS)

Santana, L.D.F.; Balan, A.

2012-01-01

Full text: The focus of our group is to determinate the role of ABC transporters in the physiology and growth of Xanthomonas citri, a phytopathogenic bacteria that infects citrus plants causing significant losses for the economy. One of the ABC transporters identified in the X. citri genome and that was showed to be active during the infection in Citrus sinensis plants was the putrescine transporter. This transporter consists of two internal membrane proteins PotG and PotH that form a pore, a cytoplasmic protein that gives energy for the transport and the periplasmic-binding protein PotF, which is responsible for the affinity and specificity of the system. Its function is associated to the microbial carcinogenesis, biofilm formation, escape from phagolysosomes, bacteriocin production, toxin activity and protection from oxidative and acid stress. In this work, we show for the first time, the expression, purification, functional and structural analyses of the X. citri PotF protein. The PotF was expressed from Escherichia coli cells strain Arctic, as a 40 kDa soluble protein, after induction of IPTG for twenty four hours at thirteen deg C. Using immobilized metal affinity chromatography for purification, the protein was eluted in the fractions with 10-500 mM of imidazole. To test the folding and cability to bind putrescine, spectroscopic analyses were performed using circular dichroism and intrinsic fluorescence. The data showed that PotF suffers conformational changes in presence of ligands and in different pH, suggesting a possible interaction with the tested ligand. Moreover, based on bioinformatics studies and molecular modeling analyses, we showed that X. citri PotF is highly conserved when compared to orthologs present in other bacteria, including the residues that form the ligand-binding site. The production of PotF in a soluble and stable form will allow us to start the crystallization trials in attempt to solve its structure. (author)

Structural and functional analyses of the putrescine binding protein PotF from Xanthomonas citri

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Santana, L.D.F.; Balan, A. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil)

2012-07-01

Full text: The focus of our group is to determinate the role of ABC transporters in the physiology and growth of Xanthomonas citri, a phytopathogenic bacteria that infects citrus plants causing significant losses for the economy. One of the ABC transporters identified in the X. citri genome and that was showed to be active during the infection in Citrus sinensis plants was the putrescine transporter. This transporter consists of two internal membrane proteins PotG and PotH that form a pore, a cytoplasmic protein that gives energy for the transport and the periplasmic-binding protein PotF, which is responsible for the affinity and specificity of the system. Its function is associated to the microbial carcinogenesis, biofilm formation, escape from phagolysosomes, bacteriocin production, toxin activity and protection from oxidative and acid stress. In this work, we show for the first time, the expression, purification, functional and structural analyses of the X. citri PotF protein. The PotF was expressed from Escherichia coli cells strain Arctic, as a 40 kDa soluble protein, after induction of IPTG for twenty four hours at thirteen deg C. Using immobilized metal affinity chromatography for purification, the protein was eluted in the fractions with 10-500 mM of imidazole. To test the folding and cability to bind putrescine, spectroscopic analyses were performed using circular dichroism and intrinsic fluorescence. The data showed that PotF suffers conformational changes in presence of ligands and in different pH, suggesting a possible interaction with the tested ligand. Moreover, based on bioinformatics studies and molecular modeling analyses, we showed that X. citri PotF is highly conserved when compared to orthologs present in other bacteria, including the residues that form the ligand-binding site. The production of PotF in a soluble and stable form will allow us to start the crystallization trials in attempt to solve its structure. (author)

Infection of an Insect Vector with a Bacterial Plant Pathogen Increases Its Propensity for Dispersal

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Coy, Monique R.; Stelinski, Lukasz L.; Pelz-Stelinski, Kirsten S.

2015-01-01

The spread of vector-transmitted pathogens relies on complex interactions between host, vector and pathogen. In sessile plant pathosystems, the spread of a pathogen highly depends on the movement and mobility of the vector. However, questions remain as to whether and how pathogen-induced vector manipulations may affect the spread of a plant pathogen. Here we report for the first time that infection with a bacterial plant pathogen increases the probability of vector dispersal, and that such movement of vectors is likely manipulated by a bacterial plant pathogen. We investigated how Candidatus Liberibacter asiaticus (CLas) affects dispersal behavior, flight capacity, and the sexual attraction of its vector, the Asian citrus psyllid (Diaphorina citri Kuwayama). CLas is the putative causal agent of huanglongbing (HLB), which is a disease that threatens the viability of commercial citrus production worldwide. When D. citri developed on CLas-infected plants, short distance dispersal of male D. citri was greater compared to counterparts reared on uninfected plants. Flight by CLas-infected D. citri was initiated earlier and long flight events were more common than by uninfected psyllids, as measured by a flight mill apparatus. Additionally, CLas titers were higher among psyllids that performed long flights than psyllid that performed short flights. Finally, attractiveness of female D. citri that developed on infected plants to male conspecifics increased proportionally with increasing CLas bacterial titers measured within female psyllids. Our study indicates that the phytopathogen, CLas, may manipulate movement and mate selection behavior of their vectors, which is a possible evolved mechanism to promote their own spread. These results have global implications for both current HLB models of disease spread and control strategies. PMID:26083763

Infection of an Insect Vector with a Bacterial Plant Pathogen Increases Its Propensity for Dispersal.

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Xavier Martini

Full Text Available The spread of vector-transmitted pathogens relies on complex interactions between host, vector and pathogen. In sessile plant pathosystems, the spread of a pathogen highly depends on the movement and mobility of the vector. However, questions remain as to whether and how pathogen-induced vector manipulations may affect the spread of a plant pathogen. Here we report for the first time that infection with a bacterial plant pathogen increases the probability of vector dispersal, and that such movement of vectors is likely manipulated by a bacterial plant pathogen. We investigated how Candidatus Liberibacter asiaticus (CLas affects dispersal behavior, flight capacity, and the sexual attraction of its vector, the Asian citrus psyllid (Diaphorina citri Kuwayama. CLas is the putative causal agent of huanglongbing (HLB, which is a disease that threatens the viability of commercial citrus production worldwide. When D. citri developed on CLas-infected plants, short distance dispersal of male D. citri was greater compared to counterparts reared on uninfected plants. Flight by CLas-infected D. citri was initiated earlier and long flight events were more common than by uninfected psyllids, as measured by a flight mill apparatus. Additionally, CLas titers were higher among psyllids that performed long flights than psyllid that performed short flights. Finally, attractiveness of female D. citri that developed on infected plants to male conspecifics increased proportionally with increasing CLas bacterial titers measured within female psyllids. Our study indicates that the phytopathogen, CLas, may manipulate movement and mate selection behavior of their vectors, which is a possible evolved mechanism to promote their own spread. These results have global implications for both current HLB models of disease spread and control strategies.

Plant pathogen-induced volatiles attract parasitoids to increase parasitism of an insect vector

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Xavier eMartini

2014-05-01

Full Text Available Interactions between plant pathogens and arthropods have been predominantly studied through the prism of herbivorous arthropods. Currently, little is known about the effect of plant pathogens on the third trophic level. This question is particularly interesting in cases where pathogens manipulate host phenotype to increase vector attraction and presumably increase their own proliferation. Indeed, a predator or a parasitoid of a vector may take advantage of this manipulated phenotype to increase its foraging performance. We explored the case of a bacterial pathogen, Candidatus Liberibacter asiaticus (Las, which modifies the odors released by its host plant (citrus trees to attract its vector, the psyllid Diaphorina citri. We found that the specialist parasitoid of D. citri, Tamarixia radiata, was attracted more toward Las-infected than uninfected plants. We demonstrated that this attractiveness was due to the release of methyl salicylate. Parasitization of D. citri nymphs on Las-infected plants was higher than on uninfected controls. Also, parasitization was higher on uninfected plants baited with methyl salicylate than on non-baited controls. This is the first report of a parasitoid ‘eavesdropping’ on a plant volatile induced by bacterial pathogen infection, which also increases effectiveness of host seeking behavior of its herbivorous vector.

Involvement of Three Esterase Genes from Panonychus citri (McGregor in Fenpropathrin Resistance

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Xiao-Min Shen

2016-08-01

Full Text Available The citrus red mite, Panonychus citri (McGregor, is a major citrus pest with a worldwide distribution and an extensive record of pesticide resistance. However, the underlying molecular mechanism associated with fenpropathrin resistance in this species have not yet been reported. In this study, synergist triphenyl phosphate (TPP dramatically increased the toxicity of fenpropathrin, suggesting involvement of carboxylesterases (CarEs in the metabolic detoxification of this insecticide. The subsequent spatiotemporal expression pattern analysis of PcE1, PcE7 and PcE9 showed that three CarEs genes were all over-expressed after insecticide exposure and higher transcripts levels were observed in different field resistant strains of P. citri. Heterologous expression combined with 3-(4,5-dimethyl-thiazol-2-yl-2,5-diphenyltetra-zolium bromide (MTT cytotoxicity assay in Spodoptera frugiperda (Sf9 cells revealed that PcE1-, PcE7- or PcE9-expressing cells showed significantly higher cytoprotective capability than parental Sf9 cells against fenpropathrin, demonstrating that PcEs probably detoxify fenpropathrin. Moreover, gene silencing through the method of leaf-mediated dsRNA feeding followed by insecticide bioassay increased the mortalities of fenpropathrin-treated mites by 31% (PcE1, 27% (PcE7 and 22% (PcE9, respectively, after individual PcE gene dsRNA treatment. In conclusion, this study provides evidence that PcE1, PcE7 and PcE9 are functional genes mediated in fenpropathrin resistance in P. citri and enrich molecular understanding of CarEs during the resistance development of the mite.

Genetic diversity of Xanthomonas axonopodis pv. citri based on plasmid profile and pulsed field gel electrophoresis

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Carvalho Flávia Maria de Souza

2005-01-01

Full Text Available Xanthomonas axonopodis pv. citri strains that cause disease in citrus were investigated by pulsed field and plasmid profile analysis. For the first method, genomic DNA was digested by the rare-cutting enzymes Xba I and Vsp I. The strains evaluated were collected in seven different States of Brazil and in Argentina, Bolivia, Paraguay and Uruguay. Genetic variability was found among strains of X. axonopodis pv. citri from different geographical areas Argentina, Bolivia and Uruguay, with similarities varying from 0.62 to 0.83. However, the strains collected in Brazil, despite being from different States, have shown a genetic similarity ranging from 0.83 to 1.00. Cluster analysis showed a relationship between genomic similarity and geographical origin of the strains. Plasmids were observed in all strains, with a total of five different plasmids, with sizes between 57.7 and 83.0 kilobases. The 72.6 kb plasmid was the most frequent, present in 15 out of 22 strains, while the 68.1 kb plasmid was observed in two strains only. Although the plasmid diversity detected in the present study was not very great, the X. axonopodis pv. citri strains evaluated showed a considerable degree of diversity with regard to this extrachromosomal genetic element.

Induction of mutations in citrus for the development of resistance to Xanthomonas citri (Hasse) Dowson

International Nuclear Information System (INIS)

Zubrzycki, H.M.; Diamante de Zubrzycki, A.

1982-01-01

With the aim of obtaining resistance to Xanthomonas citri in citrus, mutagenic treatment was carried out with X-rays and gamma rays on buds and seedlings. As a prerequisite, attempts were made to determine some physiological and structural differences in leaves in order to evaluate and analyse the reaction to the bacterium in qualitative and quantitative terms. Citrus were found to present 3-4 annual sproutings, each having a different intensity. Grapefruit showed a greater initial increase and final length than orange, but lemon growth was greater than all the others. According to their growth, leaves clustered in three groups. Those in the middle part of the sprout showed the greatest daily increase, followed in decreasing order by those of the base and those of the apex. In orange cultivars with different degrees of reaction to Xanthomonas citri a direct association between daily growth of leaves and susceptibility was found. In fully developed leaves the number of stomata per surface unit presented an inverse ratio with susceptibility. It was observed that in a given genotype the stomata frequency per surface unit was higher in young leaves than in developed ones. Also a direct association between the leaf area and the number of stomata per leaf was found. Natural field infections were measured in leaves of developed plants. Infection in each of the 3-4 annual sproutings depends on environmental factors during the growth period and the predisposition period of the leaves. The available variation for reaction to Xanthomonas citri was higher in oranges than in grapefruit. The abscission of each leaf depends on its degree of infection, but the cultivars classed as more resistant are those requiring a smaller affected area to provoke the leaf abscission. However, abscission of a leaf is independent of the fate of the neighbouring ones. The existence of a continuous grading of infection damage within the orange cultivars suggests a genetic component for resistance

A Xanthomonas citri subsp citri hypothetical protein related to virulence contains a non-functional HD domain and is implicated in flagellar motility.

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Vieira, F C F; Gonçalves, A M; Mendoza, E F R; Ferreira, R M; Costa, M L M; Balbuena, T S; Sebinelli, H G; Ciancaglini, P; Pizauro Junior, J M; Ferro, J A

2017-08-31

Citrus canker, caused by the Gram-negative bacterium Xanthomonas citri subsp citri (Xac), severely affects most economically important citrus varieties worldwide. A previous study showed that disruption of the ORF XAC1201 from the Xac 306 strain by transposon Tn5 decreased bacterium virulence in the Rangpur lime host (Citrus limonia L. Osbeck). However, little is known regarding the possible function of the hypothetical protein XAC1201 and how it affects the virulence of Xac 306. Here, we confirmed that disruption of ORF XAC1201 reduces Xac 306 virulence in two different hosts, delaying the onset of typical symptoms. In silico analysis suggested that XAC1201 interacts with the flagellar proteins FliM and FliL, known to be an important factor for virulence. In fact, motility assays revealed that the XAC1201 mutant has a significant difference in motility compared to the wild-type Xac 306. Also, a 3-D structure model revealed modified cofactor binding sites and suggested that XAC1201 has a non-functional HD domain. This hypothesis was confirmed by enzymatic assays performed in purified, XAC1201 recombinant protein expressed in Escherichia coli, which revealed no significant activities previously associated with HD domains for the tested substrates. Thus, the role of the XAC1201 protein in Xac 306 virulence seems to be related to flagellar motility, although a non-classic role for the HD domain cannot be dismissed.

Responses of the Asian citrus psyllid to volatiles emitted by the flushing shoots of its rutaceous host plants.

Science.gov (United States)

Patt, J M; Sétamou, M

2010-04-01

Diaphorina citri Kuwayama (Hemiptera: Psyllidae) carries Candidatus liberibacter spp., the putative causal agents of Huanglongbing. D. citri reproduces and develops only on the flushing shoots of its rutaceous host plants. Here we examined whether D. citri is attracted to host plant odors and a mixture of synthetic terpenes. Tests conducted in a vertically oriented Y-tube olfactometer showed that both males and females preferentially entered the Y-tube arm containing the odor from the young shoots of Murraya paniculata (L.) Jack and Citrus limon L. Burm. f. cultivar Eureka. Only males exhibited a preference for the odor of C. sinensis L., whereas the odor of C. x paradisi MacFadyen cultivar Rio Red was not attractive to both sexes. The volatiles emitted by young shoots of grapefruit cultivar Rio Red, Meyer lemon (Citrus x limon L. Burm.f.), and M. paniculata were analyzed by gas chromatograph-mass spectrometry. The samples were comprised of monoterpenes, monoterpene esters, and sesquiterpenes. The number of compounds present varied from 2 to 17, whereas the total amount of sample collected over 6 h ranged from 5.6 to 119.8 ng. The quantitatively dominant constituents were (E)-beta-ocimene, linalool, linalyl acetate, and beta-caryophyllene. The attractiveness of a mixture of synthetic terpenes, modeled on the volatiles collected from M. paniculata, was evaluated in screened cages in a no-choice test. At three observation intervals, significantly more individuals were trapped on white targets scented with the mixture than on unscented targets. These results indicate the feasibility of developing D. citri attractants patterned on actual host plant volatiles.

Behavior of bacteriome symbionts during transovarial transmission and development of the Asian citrus psyllid.

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Hiroki Dan

Full Text Available The Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Liviidae is a serious pest worldwide, transmitting Candidatus Liberibacter spp. (Alphaproteobacteria, the causative agents of a devastating citrus disease known as huanglongbing or greening disease. In a symbiotic organ called the bacteriome, D. citri possesses an organelle-like defensive symbiont, Candidatus Profftella armatura (Betaproteobacteria, and a nutritional symbiont, Ca. Carsonella ruddii (Gammaproteobacteria. Drastically reduced symbiont genomes and metabolic complementarity among the symbionts and D. citri indicate their mutually indispensable association. Moreover, horizontal gene transfer between the Profftella and Liberibacter lineages suggests ecological and evolutionary interactions between the bacteriome symbiont and the HLB pathogen. Using fluorescence in situ hybridization, we examined the behavior of Profftella and Carsonella during transovarial transmission and the development of D. citri. In the bacteriomes of sexually-mature female adults, symbionts transformed from an extremely elongated tubular form into spherical or short-rod forms, which migrated toward the ovary. The symbionts then formed mosaic masses, which entered at the posterior pole of the vitellogenic oocytes. After anatrepsis, Carsonella and Profftella migrated to the central and peripheral parts of the mass, respectively. Following the appearance of host nuclei, the mass cellularized, segregating Carsonella and Profftella in the central syncytium and peripheral uninucleate bacteriocytes, respectively. Subsequently, the uninucleate bacteriocytes harboring Profftella assembled at the posterior pole, while the syncytium, containing Carsonella, sat on the anterior side facing the germ band initiating katatrepsis. During dorsal closure, the syncytium was divided into uninuclear bacteriocytes, which surrounded the mass of bacteriocytes containing Profftella. Once fully surrounded, the bacteriocyte mass

Comparative proteomic analysis of Xanthomonas citri ssp. citri periplasmic proteins reveals changes in cellular envelope metabolism during in vitro pathogenicity induction.

Science.gov (United States)

Artier, Juliana; da Silva Zandonadi, Flávia; de Souza Carvalho, Flávia Maria; Pauletti, Bianca Alves; Leme, Adriana Franco Paes; Carnielli, Carolina Moretto; Selistre-de-Araujo, Heloisa Sobreiro; Bertolini, Maria Célia; Ferro, Jesus Aparecido; Belasque Júnior, José; de Oliveira, Julio Cezar Franco; Novo-Mansur, Maria Teresa Marques

2018-01-01

Citrus canker is a plant disease caused by Gram-negative bacteria from the genus Xanthomonas. The most virulent species is Xanthomonas citri ssp. citri (XAC), which attacks a wide range of citrus hosts. Differential proteomic analysis of the periplasm-enriched fraction was performed for XAC cells grown in pathogenicity-inducing (XAM-M) and pathogenicity-non-inducing (nutrient broth) media using two-dimensional electrophoresis combined with liquid chromatography-tandem mass spectrometry. Amongst the 40 proteins identified, transglycosylase was detected in a highly abundant spot in XAC cells grown under inducing condition. Additional up-regulated proteins related to cellular envelope metabolism included glucose-1-phosphate thymidylyltransferase, dTDP-4-dehydrorhamnose-3,5-epimerase and peptidyl-prolyl cis-trans-isomerase. Phosphoglucomutase and superoxide dismutase proteins, known to be involved in pathogenicity in other Xanthomonas species or organisms, were also detected. Western blot and quantitative real-time polymerase chain reaction analyses for transglycosylase and superoxide dismutase confirmed that these proteins were up-regulated under inducing condition, consistent with the proteomic results. Multiple spots for the 60-kDa chaperonin and glyceraldehyde-3-phosphate dehydrogenase were identified, suggesting the presence of post-translational modifications. We propose that substantial alterations in cellular envelope metabolism occur during the XAC infectious process, which are related to several aspects, from defence against reactive oxygen species to exopolysaccharide synthesis. Our results provide new candidates for virulence-related proteins, whose abundance correlates with the induction of pathogenicity and virulence genes, such as hrpD6, hrpG, hrpB7, hpa1 and hrpX. The results present new potential targets against XAC to be investigated in further functional studies. © 2016 BSPP AND JOHN WILEY & SONS LTD.

Comparison of DNA extraction methods for detection of citrus huanglongbing in Colombia

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Jorge Evelio Ángel

2014-04-01

Full Text Available Four DNA citrus plant tissue extraction protocols and three methods of DNA extraction from vector psyllid Diaphorina citri Kuwayama (Hemiptera: Psyllidae were compared as part of the validation process and standardization for detection of huanglongbing (HLB. The comparison was done using several criterias such as integrity, purity and concentration. The best quality parameters presented in terms of extraction of DNA from plant midribs tissue of citrus, were cited by Murray and Thompson (1980 and Rodríguez et al. (2010, while for the DNA extraction from psyllid vectors of HLB, the best extraction method was suggested by Manjunath et al.(2008.

Molecular Detection of Spiroplasma Citri Associated with Stubborn Disease in Citrus Orchards in Syria

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Spiroplasma citri, a phloem-limited pathogen, causes citrus stubborn disease (CSD) and can be transmitted from plant to plant by several species of phloem-feeding leafhoppers. CSD is an important disorder in certain warm and arid citrus-growing areas, and its agent has been recorded from several Med...

Crystallization, data collection and data processing of maltose-binding protein (MalE) from the phytopathogen Xanthomonas axonopodis pv. citri

International Nuclear Information System (INIS)

Souza, C. S.; Ferreira, L. C. S.; Thomas, L.; Barbosa, J. A. R. G.; Balan, A.

2009-01-01

The Xanthomonas axonopodis pv. citri maltose-binding protein MalE has been crystallized at 293 K using the hanging-drop vapour-diffusion method. Maltose-binding protein is the periplasmic component of the ABC transporter responsible for the uptake of maltose/maltodextrins. The Xanthomonas axonopodis pv. citri maltose-binding protein MalE has been crystallized at 293 K using the hanging-drop vapour-diffusion method. The crystal belonged to the primitive hexagonal space group P6 1 22, with unit-cell parameters a = 123.59, b = 123.59, c = 304.20 Å, and contained two molecules in the asymetric unit. It diffracted to 2.24 Å resolution

Low and Mid-Latitude Ionospheric Irregularities Studies Using TEC and Radio Scintillation Data from the CITRIS Radio Beacon Receiver in Low-Earth-Orbit

Science.gov (United States)

Siefring, C. L.; Bernhardt, P. A.; Huba, J.; Krall, J.; Roddy, P. A.

2009-12-01

Unique data on ionospheric plasma irregularities from the Naval Research Laboratory (NRL) CITRIS (Scintillation and TEC Receiver in Space) instrument will be presented. CITRIS is a multi-band receiver that recorded TEC (Total Electron Content) and radio scintillations from Low-Earth Orbit (LEO) on STPSat1. The 555+/5 km altitude 35° inclination orbit covers low and mid-latitudes. The measurements require propagation from a transmitter to a receiver through the F-region plasma. CITRIS used both 1) satellite beacons in LEO, such as the NRL CERTO (Coherent Electromagnetic Radio TOmography) beacons and 2) the global network of ground-based DORIS (Doppler Orbitography and Radiopositioning Integrated by Satellite) beacons. The TEC measurements allow for tracking of ionospheric disturbances and irregularities while the measurements of scintillations can simultaneously characterize their effects. CITRIS was operated in a complementary fashion with the C/NOFS (Communication/Navigations Outages Forecasting System) satellite during most of its first year of operations. C/NOFS carries a three-frequency 150/400/1067 MHz CERTO beacon and is dedicated to the study of Spread-F. In the case of Spread-F, ionospheric irregularities start with large scale size density gradients (100s of km) and cascade through complex processes to short scale sizes (10s of meters). It is typically the 100m-1km scale features that harm communication and navigation systems through scintillations. A multi-sensor approach is needed to completely understand this complex system, such as, the combination of CITRIS remote radio sensing and C/NOFS in-situ data. Several types of irregularities have been studied including Spread-F and the newly discovered dawn-side depletions. Comparisons with the physics based SAMI3 model are being performed to help our understanding of the morphology of the irregularities.

Sclerenchymatous ring as a barrier to phloem feeding by Asian citrus psyllid: Evidence from electrical penetration graph and visualization of stylet pathways.

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Justin George

Full Text Available Asian citrus psyllid (Diaphorina citri feeding behaviors play a significant role in the transmission of the phloem-limited Candidatus Liberibacter asiaticus (CLas bacterium that causes the economically devastating citrus greening disease. Sustained phloem ingestion by D. citri on CLas infected plants is required for pathogen acquisition and transmission. Recent studies have shown a fibrous ring of thick-walled sclerenchyma around the phloem in mature, fully expanded citrus leaves that is more prominent on the abaxial compared with the adaxial side. The composition and thickness of this fibrous ring may have an important role in selection of feeding sites by D. citri based on leaf age and leaf surface, which in turn can affect pathogen acquisition and transmission. We measured feeding behavior using electrical penetration graph (EPG recordings of individual D. citri adults placed on abaxial or adaxial surfaces of young or mature Valencia orange leaves to study the role of the sclerenchymatous ring in modifying D. citri feeding behavior. Feeding sites on the same leaf tissues were then sectioned and examined by epifluorescence microscopy. The duration of phloem ingestion (E2 waveform by psyllids was significantly reduced on mature compared with young leaves, and on abaxial compared with adaxial leaf surfaces. The longest duration of phloem ingestion was observed from psyllids placed on the adaxial side of young leaves that had the least developed sclerenchyma. Bouts of phloem salivation (E1 waveform, however, were significantly longer on mature leaves compared with young leaves. D. citri adults made consecutive phloem feeding attempts (bouts on the abaxial side of mature leaves and those bouts resulted in unsuccessful or shorter periods of phloem ingestion. Adults also made more frequent and longer bouts of xylem ingestion on mature leaves compared with adult psyllids placed on young leaves. Epifluorescence microscopy showed that the fibrous ring

Sclerenchymatous ring as a barrier to phloem feeding by Asian citrus psyllid: Evidence from electrical penetration graph and visualization of stylet pathways

Science.gov (United States)

George, Justin; Ammar, El-Desouky; Hall, David G.

2017-01-01

Asian citrus psyllid (Diaphorina citri) feeding behaviors play a significant role in the transmission of the phloem-limited Candidatus Liberibacter asiaticus (CLas) bacterium that causes the economically devastating citrus greening disease. Sustained phloem ingestion by D. citri on CLas infected plants is required for pathogen acquisition and transmission. Recent studies have shown a fibrous ring of thick-walled sclerenchyma around the phloem in mature, fully expanded citrus leaves that is more prominent on the abaxial compared with the adaxial side. The composition and thickness of this fibrous ring may have an important role in selection of feeding sites by D. citri based on leaf age and leaf surface, which in turn can affect pathogen acquisition and transmission. We measured feeding behavior using electrical penetration graph (EPG) recordings of individual D. citri adults placed on abaxial or adaxial surfaces of young or mature Valencia orange leaves to study the role of the sclerenchymatous ring in modifying D. citri feeding behavior. Feeding sites on the same leaf tissues were then sectioned and examined by epifluorescence microscopy. The duration of phloem ingestion (E2 waveform) by psyllids was significantly reduced on mature compared with young leaves, and on abaxial compared with adaxial leaf surfaces. The longest duration of phloem ingestion was observed from psyllids placed on the adaxial side of young leaves that had the least developed sclerenchyma. Bouts of phloem salivation (E1 waveform), however, were significantly longer on mature leaves compared with young leaves. D. citri adults made consecutive phloem feeding attempts (bouts) on the abaxial side of mature leaves and those bouts resulted in unsuccessful or shorter periods of phloem ingestion. Adults also made more frequent and longer bouts of xylem ingestion on mature leaves compared with adult psyllids placed on young leaves. Epifluorescence microscopy showed that the fibrous ring in young

Xanthomonas citri MinC Oscillates from Pole to Pole to Ensure Proper Cell Division and Shape

NARCIS (Netherlands)

Soibelmann Glock Lorenzoni, André; Dantas, Giordanni; Bergsma, Tessa; Ferreira, Henrique; Scheffers, Dirk

2017-01-01

Xanthomonas citri (Xac) is the causal agent of citrus canker, a disease that affects citrus crops and causes economic impact worldwide. To further characterize cell division in this plant pathogen, we investigated the role of the protein MinC in cell division, chromosome segregation, and

The Scintillation and Tomography Receiver in Space (CITRIS) Instrument for Ionospheric Research

Science.gov (United States)

2008-01-01

be used by data assimila- tion models to improve ionospheric density specifica- tions. Current assimilation models have been devel- oped to...geomagnetic latitude where the layer tends to be higher than at mid - latitudes . The west layer is near a mid - latitude region at –35° geomagnetic...CITRIS relative to the CERTO radio beacon transmitter on the COSMIC FM5 satellite. The orbits cross at a latitude of –35.5° N and a longitude of

Induced release of a plant-defense volatile 'deceptively' attracts insect vectors to plants infected with a bacterial pathogen.

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Rajinder S Mann

Full Text Available Transmission of plant pathogens by insect vectors is a complex biological process involving interactions between the plant, insect, and pathogen. Pathogen-induced plant responses can include changes in volatile and nonvolatile secondary metabolites as well as major plant nutrients. Experiments were conducted to understand how a plant pathogenic bacterium, Candidatus Liberibacter asiaticus (Las, affects host preference behavior of its psyllid (Diaphorina citri Kuwayama vector. D. citri were attracted to volatiles from pathogen-infected plants more than to those from non-infected counterparts. Las-infected plants were more attractive to D. citri adults than non-infected plants initially; however after feeding, psyllids subsequently dispersed to non-infected rather than infected plants as their preferred settling point. Experiments with Las-infected and non-infected plants under complete darkness yielded similar results to those recorded under light. The behavior of psyllids in response to infected versus non-infected plants was not influenced by whether or not they were carriers of the pathogen. Quantification of volatile release from non-infected and infected plants supported the hypothesis that odorants mediate psyllid preference. Significantly more methyl salicylate, yet less methyl anthranilate and D-limonene, was released by infected than non-infected plants. Methyl salicylate was attractive to psyllids, while methyl anthranilate did not affect their behavior. Feeding on citrus by D. citri adults also induced release of methyl salicylate, suggesting that it may be a cue revealing location of conspecifics on host plants. Infected plants were characterized by lower levels of nitrogen, phosphorus, sulfur, zinc, and iron, as well as, higher levels of potassium and boron than non-infected plants. Collectively, our results suggest that host selection behavior of D. citri may be modified by bacterial infection of plants, which alters release of

Induced Release of a Plant-Defense Volatile ‘Deceptively’ Attracts Insect Vectors to Plants Infected with a Bacterial Pathogen

Science.gov (United States)

Mann, Rajinder S.; Ali, Jared G.; Hermann, Sara L.; Tiwari, Siddharth; Pelz-Stelinski, Kirsten S.; Alborn, Hans T.; Stelinski, Lukasz L.

2012-01-01

Transmission of plant pathogens by insect vectors is a complex biological process involving interactions between the plant, insect, and pathogen. Pathogen-induced plant responses can include changes in volatile and nonvolatile secondary metabolites as well as major plant nutrients. Experiments were conducted to understand how a plant pathogenic bacterium, Candidatus Liberibacter asiaticus (Las), affects host preference behavior of its psyllid (Diaphorina citri Kuwayama) vector. D. citri were attracted to volatiles from pathogen-infected plants more than to those from non-infected counterparts. Las-infected plants were more attractive to D. citri adults than non-infected plants initially; however after feeding, psyllids subsequently dispersed to non-infected rather than infected plants as their preferred settling point. Experiments with Las-infected and non-infected plants under complete darkness yielded similar results to those recorded under light. The behavior of psyllids in response to infected versus non-infected plants was not influenced by whether or not they were carriers of the pathogen. Quantification of volatile release from non-infected and infected plants supported the hypothesis that odorants mediate psyllid preference. Significantly more methyl salicylate, yet less methyl anthranilate and D-limonene, was released by infected than non-infected plants. Methyl salicylate was attractive to psyllids, while methyl anthranilate did not affect their behavior. Feeding on citrus by D. citri adults also induced release of methyl salicylate, suggesting that it may be a cue revealing location of conspecifics on host plants. Infected plants were characterized by lower levels of nitrogen, phosphorus, sulfur, zinc, and iron, as well as, higher levels of potassium and boron than non-infected plants. Collectively, our results suggest that host selection behavior of D. citri may be modified by bacterial infection of plants, which alters release of specific headspace

Citrus huanglongbing: validation of Real-Time PCR (qPCR for the detection of Candidatus Liberibacter asiaticus and Candidatus Liberibacter americanus in Colombia

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Jorge Evelio Ángel

2014-12-01

Full Text Available Citrus huanglongbing (HLB is the most destructive citrus disease. Two of the three known HLB-associated Candidatus Liberibacter species were recently found to be present in the Americas. In this study, eggs, nymphs and adults of Diaphorina citri Kuwayama (Hemiptera: Liviidae and suspect citrus plant materials were collected in 25 municipalities in the departments of Cundinamarca, Santander, Valle del Cauca, Meta and Quindio (Colombia. The detection sensitivity, specificity and assay performance of the 16S rDNA-based real-time PCR (qPCR were validated for the field survey of the disease in Colombia. The validation confirmed the reliability and robustness of the real-time PCR method for the detection of HLB bacteria in host citrus plant tissues and the vector D. citri. The diagnosis was performed for Candidatus Liberibacter asiaticus (Ca. L. asiaticus and for Candidatus Liberibacter americanus (Ca. L. americanus on 168 citrus plant material samples and 239 insect samples. Neither Ca. L. asiaticus nor Ca. L. americanus were detected in the host plants or insects vector, confirming the absence of the disease in the citrus-producing areas of Colombia.

Crystallization, data collection and phasing of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri

International Nuclear Information System (INIS)

Santacruz, C. P.; Balan, A.; Ferreira, L. C. S.; Barbosa, J. A. R. G.

2006-01-01

The molybdate-binding protein (ModA) from X. axonopodis pv. citri was crystallized with sodium molybdate in the presence of PEG or sulfate. The crystal diffracted to a maximum resolution of 1.7 Å and belongs to the orthorhombic space group C222 1, with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. Xanthomonas axonopodis pv. citri ModA protein is the ABC periplasmic binding component responsible for the capture of molybdate. The protein was crystallized with sodium molybdate using the hanging-drop vapour-diffusion method in the presence of PEG or sulfate. X-ray diffraction data were collected to a maximum resolution of 1.7 Å using synchrotron radiation. The crystal belongs to the orthorhombic space group C222 1 , with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. The crystal structure was solved by molecular-replacement methods and structure refinement is in progress

Crystallization, data collection and phasing of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri

Energy Technology Data Exchange (ETDEWEB)

Santacruz, C. P.; Balan, A.; Ferreira, L. C. S. [Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, São Paulo, SP (Brazil); Barbosa, J. A. R. G., E-mail: joao@lnls.br [Centro de Biologia Molecular e Estrutural (CeBiMe), Laboratório Nacional de Luz Síncrotron (LNLS), CP 6192, Campinas, SP 13084-971 (Brazil); Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, São Paulo, SP (Brazil)

2006-03-01

The molybdate-binding protein (ModA) from X. axonopodis pv. citri was crystallized with sodium molybdate in the presence of PEG or sulfate. The crystal diffracted to a maximum resolution of 1.7 Å and belongs to the orthorhombic space group C222{sub 1,} with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. Xanthomonas axonopodis pv. citri ModA protein is the ABC periplasmic binding component responsible for the capture of molybdate. The protein was crystallized with sodium molybdate using the hanging-drop vapour-diffusion method in the presence of PEG or sulfate. X-ray diffraction data were collected to a maximum resolution of 1.7 Å using synchrotron radiation. The crystal belongs to the orthorhombic space group C222{sub 1}, with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. The crystal structure was solved by molecular-replacement methods and structure refinement is in progress.

The dual nature of trehalose in citrus canker disease: a virulence factor for Xanthomonas citri subsp. citri and a trigger for plant defence responses

KAUST Repository

Piazza, A.

2015-03-14

Xanthomonas citri subsp. citri (Xcc) is a bacterial pathogen that causes citrus canker in susceptible Citrus spp. The Xcc genome contains genes encoding enzymes from three separate pathways of trehalose biosynthesis. Expression of genes encoding trehalose-6-phosphate synthase (otsA) and trehalose phosphatase (otsB) was highly induced during canker development, suggesting that the two-step pathway of trehalose biosynthesis via trehalose-6-phosphate has a function in pathogenesis. This pathway was eliminated from the bacterium by deletion of the otsA gene. The resulting XccΔotsA mutant produced less trehalose than the wild-type strain, was less resistant to salt and oxidative stresses, and was less able to colonize plant tissues. Gene expression and proteomic analyses of infected leaves showed that infection with XccΔotsA triggered only weak defence responses in the plant compared with infection with Xcc, and had less impact on the host plant\\'s metabolism than the wild-type strain. These results suggested that trehalose of bacterial origin, synthesized via the otsA-otsB pathway, in Xcc, plays a role in modifying the host plant\\'s metabolism to its own advantage but is also perceived by the plant as a sign of pathogen attack. Thus, trehalose biosynthesis has both positive and negative consequences for Xcc. On the one hand, it enables this bacterial pathogen to survive in the inhospitable environment of the leaf surface before infection and exploit the host plant\\'s resources after infection, but on the other hand, it is a tell-tale sign of the pathogen\\'s presence that triggers the plant to defend itself against infection. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Identification of Three Kinds of Citri Reticulatae Pericarpium Based on Deoxyribonucleic Acid Barcoding and High-performance Liquid Chromatography-diode Array Detection-electrospray Ionization/Mass Spectrometry/Mass Spectrometry Combined with Chemometric Analysis

Science.gov (United States)

Yu, Xiaoxue; Zhang, Yafeng; Wang, Dongmei; Jiang, Lin; Xu, Xinjun

2018-01-01

Background: Citri Reticulatae Pericarpium is the dried mature pericarp of Citrus reticulata Blanco which can be divided into “Chenpi” and “Guangchenpi.” “Guangchenpi” is the genuine Chinese medicinal material in Xinhui, Guangdong province; based on the greatest quality and least amount, it is most expensive among others. Hesperidin is used as the marker to identify Citri Reticulatae Pericarpium described in the Chinese Pharmacopoeia 2010. However, both “Chenpi” and “Guangchenpi” contain hesperidin so that it is impossible to differentiate them by measuring hesperidin. Objective: Our study aims to develop an efficient and accurate method to separate and identify “Guangchenpi” from other Citri Reticulatae Pericarpium. Materials and Methods: The genomic deoxyribonucleic acid (DNA) of all the materials was extracted and then the internal transcribed spacer 2 was amplified, sequenced, aligned, and analyzed. The secondary structures were created in terms of the database and website established by Jörg Schultz et al. High-performance liquid chromatography-diode array detection-electrospray Ionization/mass spectrometry (HPLC-DAD-ESI-MS)/MS coupled with chemometric analysis was applied to compare the differences in chemical profiles of the three kinds of Citri Reticulatae Pericarpium. Results: A total of 22 samples were classified into three groups. The results of DNA barcoding were in accordance with principal component analysis and hierarchical cluster analysis. Eight compounds were deduced from HPLC-DAD-ESI-MS/MS. Conclusions: This method is a reliable and effective tool to differentiate the three Citri Reticulatae Pericarpium. SUMMARY The internal transcribed spacer 2 regions and the secondary structure among three kinds of Citri Reticulatae Pericarpium varied considerablyAll the 22 samples were analyzed by high-performance liquid chromatography (HPLC) to obtain the chemical profilesPrincipal component analysis and hierarchical cluster analysis

PENDIDIKAN DAN PELATIHAN PENGENDALIAN TERPADU PENYAKIT CVPD (CITRUS VEIN PHLOEM DEGENERATION PADA PERTANAMAN JERUK DI DESA PELAGA, KECAMATAN PETANG, KABUPATEN BADUNG

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WAYAN ADIARTAYASA

2012-09-01

Full Text Available ABSTRACT Education and training on integrated pest management (IPM of CVPD (Citrus Vein Phloem Degeneration was held at Pelaga village, Badung regency on 22 September 2005. The activities aimed to improve knowledge of the farmers about CVPD and factors influencing the intensity, insect vector and integrated pest management of CVPD. The topics convered were disease symptons, and pathogen bacteria, Diaphorina citri are as vector insect infection mechanism, disease distribubution and integrated management of CVPD. The training was attended by 26 participants from local groups of Pelaga village. The method used in activities were lectures, demonstration and practical work in the citrus field. All participants enthusiastically took part and hoped to have the next intensive traning of citrus culture.

Modelling the effect of an alternative host population on the spread of citrus Huanglongbing

Science.gov (United States)

d'A. Vilamiu, Raphael G.; Ternes, Sonia; Laranjeira, Francisco F.; de C. Santos, Tâmara T.

2013-10-01

The objective of this work was to model the spread of citrus Huanglongbing (HLB) considering the presence of a population of alternative hosts (Murraya paniculata). We developed a compartmental deterministic mathematical model for representing the dynamics of HLB disease in a citrus orchard, including delays in the latency and incubation phases of the disease in the plants and a delay period on the nymphal stage of Diaphorina citri, the insect vector of HLB in Brazil. The results of numerical simulations indicate that alternative hosts should not play a crucial role on HLB dynamics considering a typical scenario for the Recôncavo Baiano region in Brazil . Also, the current policy of removing symptomatic plants every three months should not be expected to significantly hinder HLB spread.

Thermal hygrometric requirements for the rearing and release of Tamarixia radiata (Waterston (Hymenoptera, Eulophidae

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Mariuxi Lorena Gómez-Torres

2014-09-01

Full Text Available Thermal hygrometric requirements for the rearing and release of Tamarixia radiata (Waterston (Hymenoptera, Eulophidae. Tamarixia radiata is the main agent for the biological control of Diaphorina citri in Brazil with a parasitism rate ranging from 20 to 80%. This study investigated the influence of temperature on the development, fecundity and longevity of adults of T. radiata and the effect of relative humidity (RH on their parasitism capacity and survival rate in the pre-imaginal period. The effect of temperature was assessed in the range between 15 and 35 ± 1ºC, 70 ± 10% RH, and a 14-h photophase. The RH effect was evaluated in the range from 30 to 90 ± 10%, temperature at 25 ± 1ºC, and photophase of 14-h. At 25ºC, circa 166.7 nymphs were parasitized, the highest parasitism capacity observed compared to other treatments. The longest longevity of females was observed at 25ºC, although the rate did not differ in the 20-30ºC temperature range. The threshold temperature (TT was 7.2ºC, and 188.7 degrees-day were required for the development (egg-to-adult period. The parasitism rate and longevity were higher at 50 and 70% of RH. This shows that temperature and RH may affect the parasitism capacity of T. radiata on nymphs of D. citri, which can explain the great parasitism variation for D. citri observed in citrus groves in São Paulo State, Brazil.

Synthetic blends of volatile, phytopathogen-induced odorants can be used to manipulate vector behavior.

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Alexander eAksenov

2014-12-01

Full Text Available Volatile organic compounds (VOCs are emitted from all plants and these VOCs are important means of communication between plants and insects. It has been documented that pathogen infections alter VOC profiles rendering infected plants more attractive to specific vectors transmitting these pathogens than uninfected plants, thus potentially aiding in pathogen propagation. Mimicking these chemical cues might enable insect attraction away from the plant or disruption of host finding behavior of the vector. However, the practical implications have not been fully explored. We used citrus, Diaphorina citri and huanglongbing (HLB as a model host-vector-disease system because HLB threatens citrus production worldwide and is similar to other critical diseases of food crops, such as Zebra Chip affecting potato. We formulated a synthetic chemical blend using selected HLB-specific biomarker compounds, and tested the blend with the Attenu assay system for chemosensory proteins. The Attenu assay system is a procedure that identifies interactions between insect chemosensory proteins and their ligands. We found that an equimolar mixture of compounds mimicking the volatile profile of HLB-infected citrus bound chemosensory proteins. Further investigation of this blend in laboratory behavioral assays resulted in development of a synthetic lure that was more attractive to D. citri than natural citrus tree volatiles. This strategy could provide a new route to produce chemical lures for vector population control for a variety of plant and/or animal systems and it may result in the development of a practical lure for monitoring vectors of disease, such as D. citri.

Diel periodicity of pheromone release by females of Planococcus citri and Planococcus ficus and the temporal flight activity of their conspecific males

Science.gov (United States)

Levi-Zada, Anat; Fefer, Daniela; David, Maayan; Eliyahu, Miriam; Franco, José Carlos; Protasov, Alex; Dunkelblum, Ezra; Mendel, Zvi

2014-08-01

The diel periodicity of sex pheromone release was monitored in two mealybug species, Planococcus citri and Planococcus ficus (Hemiptera; Pseudococcidae), using sequential SPME/GCMS analysis. A maximal release of 2 ng/h pheromone by 9-12-day-old P. citri females occurred 1-2 h before the beginning of photophase. The highest release of pheromone by P. ficus females was 1-2 ng/2 h of 10-20-day-old females, approximately 2 h after the beginning of photophase. Mating resulted in termination of the pheromone release in both mealybug species. The temporal flight activity of the males was monitored in rearing chambers using pheromone baited delta traps. Males of both P. citri and P. ficus displayed the same flight pattern and began flying at 06:00 hours when the light was turned on, reaching a peak during the first and second hour of the photophase. Our results suggest that other biparental mealybug species display also diel periodicities of maximal pheromone release and response. Direct evaluation of the diel periodicity of the pheromone release by the automatic sequential analysis is convenient and will be very helpful in optimizing the airborne collection and identification of other unknown mealybug pheromones and to study the calling behavior of females. Considering this behavior pattern may help to develop more effective pheromone-based management strategies against mealybugs.

Crystallographic structure and substrate-binding interactions of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri.

Science.gov (United States)

Balan, Andrea; Santacruz-Pérez, Carolina; Moutran, Alexandre; Ferreira, Luís Carlos Souza; Neshich, Goran; Gonçalves Barbosa, João Alexandre Ribeiro

2008-02-01

In Xanthomonas axonopodis pv. citri (Xac or X. citri), the modA gene codes for a periplasmic protein (ModA) that is capable of binding molybdate and tungstate as part of the ABC-type transporter required for the uptake of micronutrients. In this study, we report the crystallographic structure of the Xac ModA protein with bound molybdate. The Xac ModA structure is similar to orthologs with known three-dimensional structures and consists of two nearly symmetrical domains separated by a hinge region where the oxyanion-binding site lies. Phylogenetic analysis of different ModA orthologs based on sequence alignments revealed three groups of molybdate-binding proteins: bacterial phytopathogens, enterobacteria and soil bacteria. Even though the ModA orthologs are segregated into different groups, the ligand-binding hydrogen bonds are mostly conserved, except for Archaeglobus fulgidus ModA. A detailed discussion of hydrophobic interactions in the active site is presented and two new residues, Ala38 and Ser151, are shown to be part of the ligand-binding pocket.

First records of parasitoids attacking the Asian citrus psyllid in Ecuador

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Diego E. Portalanza

Full Text Available ABSTRACT First records of parasitoids attacking the Asian citrus psyllid in Ecuador. The objective of the current study was to investigate the presence of natural enemies of Diaphorina citri (Hemiptera: Liviidae (the Asian citrus psyllid in Ecuador. Incidence of parasitoid Diaphorencyrtus aligarhensis (Hymenoptera: Encyrtidae was assessed between November 2015 and March 2016, in Letamendi, Febres-Cordero and Tarqui, urban districts of Guayaquil. Highest incidence of parasitism occurred in those regions and seasons of the year with the highest temperatures commensurate with increase of citrus plant shoots. Similar to their host, these parasitoids appear to have established in Ecuador by accident, and were not the result of purposeful introduction. This fortuitous introduction is a potentially helpful tool in controlling the Asian citrus psyllid, and potentially Huanglongbing.

PENDIDIKAN DAN PELATIHAN PEMBIBITAN JERUK BEBAS PENYAKIT CVPD (Citrus Vein Phloem Degeneration DI DESA KATUNG KINTAMANI

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Adiartayasa W

2012-10-01

Full Text Available The community service in the form of training about free orange seed of disease CVPD (Citrus Vein Phloem Degenaration was conducted in Katung village, district of Kintamani, Bangli regency on 10 August 2007. The training activity aimed to improve knowledge of farmers about CVPD disease, symptom, transmission, insect vector, and integrated pest management. The topics covered were disease symptoms, and pathogen bacteria, Diaphorina citri are as insect vector infection mechanism, election of free orange mains crops CVPD, disease distribution and integrated management of CVPD. The training was attended by 25 participants from local groups of Katung village. The methods used in this activity were lectures, demonstration and practical work in the citrus field. All participants enthusiastically took part and hoped to have the next intensive training of citrus culture.

Insetos Sugadores (Sternorrhyncha em Cultivo Orgânico de Tangerina cv. Poncã (Citrus reticulata Blanco: Diversidade, Constância, Freqüência e Flutuação Populacional

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William Costa Rodrigues

2009-08-01

Full Text Available O objetivo do trabalho foi estudar a diversidade, constância, freqüência e a flutuação populacional de insetos sugadores da subordem Sternorrhyncha associados à tangerina cv. Poncã sob o sistema de cultivo orgânico. Os levantamentos foram realizados entre outubro de 2002 e outubro de 2003. Dentre os aleirodídeos, Aleurothrixus floccosus Maskell destacou-se dos demais, com média populacional igual a 74,06%. Aleurotrachelus cruzi Cassino obteve média de 52,6%. Entre as cochonilhas destacaram-se Selenaspidus articulatus (Morgan (23,96% e Pinnaspis aspidistrae (Signoret (19,38%. O psilídeo Diaphorina citri Kuwayama obteve média de 52,6%, sendo a segunda espécie com maior média. Os pulgões Toxoptera citricida (Kirkaldy e Toxoptera aurantii (Boyer, somente ocorreram em período de brotação e floração, com média populacional de 7,6% e 6,25%, respectivamente. Quanto à diversidade das espécies o período com maior diversidade, ocorreu na 1ª quinzena de setembro de 2003 (H’= 0,964, coincidindo com a menor dominância (D= 0,200, e menor diversidade na 1º quinzena de maio de 2003 (H’= 0,657 e maior dominância (D= 0,522. As espécies constantes foram: A. floccosus (100%, A. cruzi (96%, Coccus viridis (Green (100%, P. aspidistrae (100% e D. citri (100%. As espécies mais freqüentes foram: A. floccosus (30,79% e D. citri (21,87%. Os resultados sugerem que as espécies verificadas possuem a dinâmica populacional em parte influenciada pela competição interespecífica, pois quando há um aumento populacional dos aleirodídeos, há um decréscimo da população das cochonilhas e vice-versa, além dos fatores ambientais, a predação e o parasitismo.Sucker Insects (Sternorrhyncha in Organic Cropping of Tangerine cv. Ponkan (Citrus reticulata Blanco: Diversity, Constancy, Frequency and Populational FluctuationAbstract. The aim of this work was to study the diversity, constancy, frequency and the population fluctuation of the sucker

Horizontal gene acquisition of Liberibacter plant pathogens from a bacteriome-confined endosymbiont of their psyllid vector.

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Atsushi Nakabachi

Full Text Available he Asian citrus psyllid Diaphorina citri is a notorious agricultural pest that transmits the phloem-inhabiting alphaproteobacterial 'Candidatus Liberibacter asiaticus' and allied plant pathogens, which cause the devastating citrus disease called Huanglongbing or greening disease. D. citri harbors two distinct bacterial mutualists in the symbiotic organ called bacteriome: the betaproteobacterium 'Candidatus Profftella armatura' in the syncytial cytoplasm at the center of the bacteriome, and the gammaproteobacterium 'Candidatus Carsonella ruddii' in uninucleate bacteriocytes. Here we report that a putative amino acid transporter LysE of Profftella forms a highly supported clade with proteins of L. asiaticus, L. americanus, and L. solanacearum. L. crescens, the most basal Liberibacter lineage currently known, lacked the corresponding gene. The Profftella-Liberibacter subclade of LysE formed a clade with proteins from betaproteobacteria of the order Burkholderiales, to which Profftella belongs. This phylogenetic pattern favors the hypothesis that the Liberibacter lineage acquired the gene from the Profftella lineage via horizontal gene transfer (HGT after L. crescens diverged from other Liberibacter lineages. K A/K S analyses further supported the hypothesis that the genes encoded in the Liberibacter genomes are functional. These findings highlight the possible evolutionary importance of HGT between plant pathogens and their insect vector's symbionts that are confined in the symbiotic organ and seemingly sequestered from external microbial populations.

Acquisition, Replication and Inoculation of Candidatus Liberibacter asiaticus following Various Acquisition Periods on Huanglongbing-Infected Citrus by Nymphs and Adults of the Asian Citrus Psyllid.

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El-Desouky Ammar

Full Text Available The Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae, is the primary vector of Candidatus Liberibacter asiaticus (Las implicated as causative agent of citrus huanglongbing (citrus greening, currently the most serious citrus disease worldwide. Las is transmitted by D. citri in a persistent-circulative manner, but the question of replication of this bacterium in its psyllid vector has not been resolved. Thus, we studied the effects of the acquisition access period (AAP by nymphs and adults of D. citri on Las acquisition, multiplication and inoculation/transmission. D. citri nymphs or adults (previously non-exposed to Las were caged on Las-infected citrus plants for an AAP of 1, 7 or 14 days. These 'Las-exposed' psyllids were then transferred weekly to healthy citrus or orange jasmine plants, and sampled via quantitative polymerase chain reaction (qPCR analysis 1-42 days post-first access to diseased plants (padp; all tested nymphs became adults 7-14 days padp. Our results indicate that following 1 or 7 day AAP as nymphs 49-59% of Las-exposed psyllids became Las-infected (qPCR-positive, whereas only 8-29% of the psyllids were infected following 1-14 day AAP as adults. Q-PCR analysis also indicated that Las titer in the Las-exposed psyllids (relative to that of the psyllid S20 ribosomal protein gene was: 1 significantly higher, and increasing at a faster rate, following Las acquisition as nymphs compared to that following Las acquisition as adults; 2 higher as post-acquisition time of psyllids on healthy plants increased reaching a peak at 14-28 days padp for nymphs and 21-35 days padp for adults, with Las titer decreasing or fluctuating after that; 3 higher with longer AAP on infected plants, especially with acquisition as adults. Our results strongly suggest that Las multiplies in both nymphs and adults of D. citri but attains much higher levels in a shorter period of time post-acquisition when acquired by nymphs than when acquired by

Characterization of the sulfate uptake and assimilation pathway from Xanthomonas citri - targets for bacterial growth inhibitors

Energy Technology Data Exchange (ETDEWEB)

Tambascia, C.; Balan, A. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil)

2012-07-01

Full text: Microorganisms require sulfur for growth and obtain it either for inorganic sulfate or organosulfur compounds. ATP-Binding Cassete (SulT family) or major facilitator superfamily-type (SulP) transporters are responsible for the sulfate transport into the cell. In Xanthomonas citri, the phytopathogenic bacterium that causes the canker citrus disease, there are no reports related to the importance of these transporters during in vitro or in vivo infection. We identified in X. citri genome all the genes that belong to the well-characterized cys regulon from Escherichia coli and Salmonella typhimurium, which includes three ABC transporters and all the enzymes necessary for sulfate oxide reduction to sulfide and cysteine. Once these genes have been shown to be extremely important for bacteria growth and development in different environments, we chose the sbpcysWUA and cysDNCHIJG operons, which encodes the ABC inorganic sulfate ABC transporter and all the enzymes necessary for conversion of sulfate in cysteine, respectively. As a step for crystallization trials and resolution of their tridimensional structures, the referred genes were amplified and cloned into the cloning vector pGEM T-easy. In addition, using bioinformatics tools and molecular modeling we characterized all the protein functions as well as built tridimensional models of their structure for determination of the active sites. The importance of each protein is discussed aiming the discovery of a good target for development of inhibitors that could block the bacterium growth. (author)

Characterization of the sulfate uptake and assimilation pathway from Xanthomonas citri - targets for bacterial growth inhibitors

International Nuclear Information System (INIS)

Tambascia, C.; Balan, A.

2012-01-01

Full text: Microorganisms require sulfur for growth and obtain it either for inorganic sulfate or organosulfur compounds. ATP-Binding Cassete (SulT family) or major facilitator superfamily-type (SulP) transporters are responsible for the sulfate transport into the cell. In Xanthomonas citri, the phytopathogenic bacterium that causes the canker citrus disease, there are no reports related to the importance of these transporters during in vitro or in vivo infection. We identified in X. citri genome all the genes that belong to the well-characterized cys regulon from Escherichia coli and Salmonella typhimurium, which includes three ABC transporters and all the enzymes necessary for sulfate oxide reduction to sulfide and cysteine. Once these genes have been shown to be extremely important for bacteria growth and development in different environments, we chose the sbpcysWUA and cysDNCHIJG operons, which encodes the ABC inorganic sulfate ABC transporter and all the enzymes necessary for conversion of sulfate in cysteine, respectively. As a step for crystallization trials and resolution of their tridimensional structures, the referred genes were amplified and cloned into the cloning vector pGEM T-easy. In addition, using bioinformatics tools and molecular modeling we characterized all the protein functions as well as built tridimensional models of their structure for determination of the active sites. The importance of each protein is discussed aiming the discovery of a good target for development of inhibitors that could block the bacterium growth. (author)

Chemical composition and fungicidal activity of the essential oil of Thymus vulgaris against Alternaria citri

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Erica A. Soto Mendívil

2006-01-01

Full Text Available Se analizó químicamente el aceite esencial de tomillo (Thymus vulgaris L. por Cromatografía de Gases/Espectroscopia de Masas y se evaluó su actividad fungicida. Los principales constituyentes fueron borneol (28.4%, timol (16.6%, carvacrol metil eter (9.6%, camfeno (6.9%, α-humulene (6.4% y carvacrol (5.0%. Las pruebas de actividad fungicida (in vitro indicaron que la concentración de 1000ppm del aceite esencial de tomillo fue efectivo para inhibir a Alternaria citri, cuando se adicionó al medio de cultivo agar papa dextrosa

Citrus huanglongbing in São Paulo State, Brazil: PCR detection of the 'Candidatus' Liberibacter species associated with the disease.

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do Carmo Teixeira, Diva; Luc Danet, Jean; Eveillard, Sandrine; Cristina Martins, Elaine; de Jesus Junior, Waldir Cintra; Takao Yamamoto, Pedro; Aparecido Lopes, Silvio; Beozzo Bassanezi, Renato; Juliano Ayres, Antonio; Saillard, Colette; Bové, Joseph Marie

2005-06-01

Symptoms of huanglongbing (HLB), one of the most serious diseases of citrus in Asia and Africa, have been noticed in March 2004 in the Araraquara region of São Paulo State, Brazil. HLB has not been reported previously from America. The causal HLB bacteria, Candidatus Liberibacter africanus in Africa and Candidatus Liberibacter asiaticus in Asia, can be detected in symptomatic citrus leaves by PCR amplification of their 16S rDNA with previously described primers. When this technique was applied to 43 symptomatic leaf samples from the Araraquara region, all PCR reactions were negative. This suggested that a new pathogen, not detected by the above primers, could be involved in HLB in the State of São Paulo. Indeed, by using universal primers for amplification of bacterial 16S rDNA, a new liberibacter species, Candidatus Liberibacter americanus, has recently been identified. Specific primers for PCR amplification of the 16S rDNA of Ca. L. americanus have been selected. Using these primers, the new liberibacter could be detected in 214 symptomatic leaf samples tested. The leaves of two additional samples were infected with Candidatus Liberibacter asiaticus, and two further samples contained both Ca. L. americanus and Ca. L. asiaticus. The samples came from 47 farms in 35 municipalities. The psyllid vector of Ca. L. asiaticus, Diaphorina citri, is established in South, Central, and North America (Florida and Texas). Ca. L. americanus could be detected by PCR in several batches of D. citri psyllids collected on symptomatic sweet orange trees infected with Ca. L. americanus, strongly suggesting that D. citri is the vector of Ca. L. americanus. The results reported here confirm the presence of HLB in the State of São Paulo. Ca. L. americanus is the most widely distributed pathogen.

Multiplexed lateral flow microarray assay for detection of citrus pathogens Xylella fastidiosa and Xanthomonas axonopodis pv citri

Energy Technology Data Exchange (ETDEWEB)

Cary,; Bruce, R [Santa Fe, NM; Stubben, Christopher J [Los Alamos, NM

2011-03-22

The invention provides highly sensitive and specific assays for the major citrus pathogens Xylella fastidiosa and Xanthomonas axonopodis, including a field deployable multiplexed assay capable of rapidly assaying for both pathogens simultaneously. The assays are directed at particular gene targets derived from pathogenic strains that specifically cause the major citrus diseases of citrus variegated chlorosis (Xylella fastidiosa 9a5c) and citrus canker (Xanthomonas axonopodis pv citri). The citrus pathogen assays of the invention offer femtomole sensitivity, excellent linear dynamic range, and rapid and specific detection.

Crystallization and preliminary X-ray diffraction analysis of XAC1151, a small heat-shock protein from Xanthomonas axonopodis pv. citri belonging to the α-crystallin family

Energy Technology Data Exchange (ETDEWEB)

Hilario, Eduardo; Teixeira, Elaine Cristina; Pedroso, Gisele Audrei; Bertolini, Maria Célia [Departamento de Bioquímica e Tecnologia Química, Instituto de Química, Universidade Estadual Paulista, Araraquara-SP (Brazil); Medrano, Francisco Javier, E-mail: fjmedrano@yahoo.com [Departamento de Cristalografia de Proteínas, Centro de Biologia Molecular Estrutural, Laboratório Nacional de Luz Síncrotron, Caixa Postal 6192, CEP 13084-971, Campinas-SP (Brazil); Departamento de Bioquímica e Tecnologia Química, Instituto de Química, Universidade Estadual Paulista, Araraquara-SP (Brazil)

2006-05-01

XAC1151, a small heat-shock protein from X. axonopodis pv. citri belonging to the α-crystallin family, was crystallized using the sitting-drop vapour-diffusion method in the presence of ammonium phosphate. X-ray diffraction data were collected to 1.65 Å resolution using a synchrotron-radiation source. The hspA gene (XAC1151) from Xanthomonas axonopodis pv. citri encodes a protein of 158 amino acids that belongs to the small heat-shock protein (sHSP) family of proteins. These proteins function as molecular chaperones by preventing protein aggregation. The protein was crystallized using the sitting-drop vapour-diffusion method in the presence of ammonium phosphate. X-ray diffraction data were collected to 1.65 Å resolution using a synchrotron-radiation source. The crystal belongs to the rhombohedral space group R3, with unit-cell parameters a = b = 128.7, c = 55.3 Å. The crystal structure was solved by molecular-replacement methods. Structure refinement is in progress.

Chemical Composition, Antioxidant and Antimicrobial Activity of Pericarpium Citri Reticulatae Essential Oil

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Bei Gao

2011-05-01

Full Text Available The chemical composition, antioxidant and antimicrobial activity of Pericarpium Citri Reticulatae (PCR essential oil obtained using an improved Clevenger type apparatus were studied. Among the five different PCRs examined the highest yield of essential oil was found in Chachi 2004 (harvested and stored in 2004 and the lowest in Chachi 2008 (harvested and stored in 2008. Fifty three different volatile compounds were determined, including terpenic hydrocarbons, alcohols, aldehydes, ketones and esters. D-limonene, one of terpenes, was the major constituent in PCR. The antioxidant capacity of PCR essential oil varied considerably with the duration of storage time, and the oil from Chachi 1994 has the strongest ferric-reducing antioxidant power. In addition, the essential oil possessed varying degrees of antimicrobial activity against Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, except Streptococcus faecalis, while had no effect on Gram-negative bacteria (Escherichia coli, Enterobacter cloacae.

Simultaneous quantification of coumarins, flavonoids and limonoids in Fructus Citri Sarcodactylis by high performance liquid chromatography coupled with diode array detector.

Science.gov (United States)

Chu, Jun; Li, Song-Lin; Yin, Zhi-Qi; Ye, Wen-Cai; Zhang, Qing-Wen

2012-07-01

A high performance liquid chromatography coupled with diode array detector (HPLC-DAD) method was developed for simultaneous quantification of eleven major bioactive components including six coumarins, three flavonoids and two limonoids in Fructus Citri Sarcodactylis. The analysis was performed on a Cosmosil 5 C(18)-MS-II column (4.6 mm × 250 mm, 5 μm) with water-acetonitrile gradient elution. The method was validated in terms of linearity, sensitivity, precision, stability and accuracy. It was found that the calibration curves for all analytes showed good linearity (R(2)>0.9993) within the test ranges. The overall limit of detection (LOD) and limit of quantification (LOQ) were less than 3.0 and 10.2 ng. The relative standard deviations (RSDs) for intra- and inter-day repeatability were not more than 4.99% and 4.92%, respectively. The sample was stable for at least 48 h. The spike recoveries of eleven components were 95.1-104.9%. The established method was successfully applied to determine eleven components in three samples from different locations. The results showed that the newly developed HPLC-DAD method was linear, sensitive, precise and accurate, and could be used for quality control of Fructus Citri Sarcodactylis. Copyright © 2012 Elsevier B.V. All rights reserved.

Sobre o condicionamento alimentar na cochonilha-branca, Planococcus citri (Risso (Hemiptera: pseudococcidae Regarding to host conditioning in citrus mealybug Planococcus citri (Risso (Hemiptera: pseudococcidae

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Lenira Viana Costa Santa-Cecília

2013-03-01

Full Text Available Estudos do comportamento alimentar mediante a técnica de "Electrical Penetration Graphs" (EPG das cochonilhas-farinhentas (Pseudococcidae provenientes de um hospedeiro de criação alternativo têm mostrado que esses insetos não atingem ou demoram cerca de 9 horas para alcançar a fase floemática. Por outro lado, aqueles provenientes do hospedeiro-fonte atingem a fase floemática mais rapidamente e apresentam maior frequência de alimentação nos vasos crivados. Esses resultados indicam a presença do fenômeno de condicionamento alimentar, ainda não demonstrado em cochonilhas. Assim, o presente trabalho teve como objetivo determinar a existência desse fenômeno em Planococcus citri (Risso (Hemiptera: Pseudococcidae. Foram realizados testes de livre escolha, monitoramento eletrônico (EPG e estudos de alguns parâmetros biológicos. Em todos os experimentos, o cafeeiro (Coffea arabica L., os citros (Citrus sinensis L. e abóbora (Cucurbita maxima L. foram utilizados como substratos de criação (fonte da cochonilha, sendo os tratamentos constituídos pela combinação entre os hospedeiros-fonte e os hospedeiros receptores (café e citros. O teste de escolha entre cafeeiro e citros nas primeiras 72 horas mostrou que as cochonilhas criadas em cafeeiro apresentaram preferência pelo cafeeiro; aquelas originadas dos citros mostraram uma tendência, embora não significativa, em selecionar os citros em relação ao cafeeiro e aquelas criadas em abóbora não mostraram preferência por nenhum dos hospedeiros. Os estudos do comportamento alimentar mediante o monitoramento eletrônico (EPG mostraram que a fase floemática, considerada como a fase de aceitação do hospedeiro, foi mais frequente em cafeeiro, seja com cochonilhas oriundas deste substrato, seja de citros. Aqueles insetos mantidos em abóbora e transferidos para o cafeeiro ou citros apresentaram excepcionalmente ou não apresentaram nenhuma fase floemática, respectivamente. A

Dynamical System and Nonlinear Regression for Estimate Host-Parasitoid Relationship

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Ileana Miranda Cabrera

2010-01-01

Full Text Available The complex relationships of a crop with the pest, its natural enemies, and the climate factors exist in all the ecosystems, but the mathematic models has studied only some components to know the relation cause-effect. The most studied system has been concerned with the relationship pest-natural enemies such as prey-predator or host-parasitoid. The present paper shows a dynamical system for studying the relationship host-parasitoid (Diaphorina citri, Tamarixia radiata and shows that a nonlinear model permits the estimation of the parasite nymphs using nymphs healthy as the known variable. The model showed the functional answer of the parasitoid, in which a point arrives that its density is not augmented although the number host increases, and it becomes necessary to intervene in the ecosystem. A simple algorithm is used to estimate the parasitoids level using the priori relationship between the host and the climate factors and then the nonlinear model.

The quest for a non-vector psyllid: Natural variation in acquisition and transmission of the huanglongbing pathogen ‘Candidatus Liberibacter asiaticus’ by Asian citrus psyllid isofemale lines

Science.gov (United States)

Ammar, El-Desouky; Hall, David G.; Hosseinzadeh, Saeed

2018-01-01

Genetic variability in insect vectors is valuable to study vector competence determinants and to select non-vector populations that may help reduce the spread of vector-borne pathogens. We collected and tested vector competency of 15 isofemale lines of Asian citrus psyllid, Diaphorina citri, vector of ‘Candidatus Liberibacter asiaticus’ (CLas). CLas is associated with huanglongbing (citrus greening), the most serious citrus disease worldwide. D. citri adults were collected from orange jasmine (Murraya paniculata) hedges in Florida, and individual pairs (females and males) were caged on healthy Murraya plants for egg laying. The progeny from each pair that tested CLas-negative by qPCR were maintained on Murraya plants and considered an isofemale line. Six acquisition tests on D. citri adults that were reared as nymphs on CLas-infected citrus, from various generations of each line, were conducted to assess their acquisition rates (percentage of qPCR-positive adults). Three lines with mean acquisition rates of 28 to 32%, were classified as ‘good’ acquirers and three other lines were classified as ‘poor’ acquirers, with only 5 to 8% acquisition rates. All lines were further tested for their ability to inoculate CLas by confining CLas-exposed psyllids for one week onto healthy citrus leaves (6–10 adults/leaf/week), and testing the leaves for CLas by qPCR. Mean inoculation rates were 19 to 28% for the three good acquirer lines and 0 to 3% for the three poor acquirer lines. Statistical analyses indicated positive correlations between CLas acquisition and inoculation rates, as well as between CLas titer in the psyllids and CLas acquisition or inoculation rates. Phenotypic and molecular characterization of one of the good and one of the poor acquirer lines revealed differences between them in color morphs and hemocyanin expression, but not the composition of bacterial endosymbionts. Understanding the genetic architecture of CLas transmission will enable the

Sobre o condicionamento alimentar na cochonilha-branca, Planococcus citri (Risso (Hemiptera: pseudococcidae

Directory of Open Access Journals (Sweden)

Lenira Viana Costa Santa-Cecília

2013-03-01

Full Text Available Estudos do comportamento alimentar mediante a técnica de "Electrical Penetration Graphs" (EPG das cochonilhas-farinhentas (Pseudococcidae provenientes de um hospedeiro de criação alternativo têm mostrado que esses insetos não atingem ou demoram cerca de 9 horas para alcançar a fase floemática. Por outro lado, aqueles provenientes do hospedeiro-fonte atingem a fase floemática mais rapidamente e apresentam maior frequência de alimentação nos vasos crivados. Esses resultados indicam a presença do fenômeno de condicionamento alimentar, ainda não demonstrado em cochonilhas. Assim, o presente trabalho teve como objetivo determinar a existência desse fenômeno em Planococcus citri (Risso (Hemiptera: Pseudococcidae. Foram realizados testes de livre escolha, monitoramento eletrônico (EPG e estudos de alguns parâmetros biológicos. Em todos os experimentos, o cafeeiro (Coffea arabica L., os citros (Citrus sinensis L. e abóbora (Cucurbita maxima L. foram utilizados como substratos de criação (fonte da cochonilha, sendo os tratamentos constituídos pela combinação entre os hospedeiros-fonte e os hospedeiros receptores (café e citros. O teste de escolha entre cafeeiro e citros nas primeiras 72 horas mostrou que as cochonilhas criadas em cafeeiro apresentaram preferência pelo cafeeiro; aquelas originadas dos citros mostraram uma tendência, embora não significativa, em selecionar os citros em relação ao cafeeiro e aquelas criadas em abóbora não mostraram preferência por nenhum dos hospedeiros. Os estudos do comportamento alimentar mediante o monitoramento eletrônico (EPG mostraram que a fase floemática, considerada como a fase de aceitação do hospedeiro, foi mais frequente em cafeeiro, seja com cochonilhas oriundas deste substrato, seja de citros. Aqueles insetos mantidos em abóbora e transferidos para o cafeeiro ou citros apresentaram excepcionalmente ou não apresentaram nenhuma fase floemática, respectivamente. A

Suppression of greasy spot disease caused by Mycosphaerella citri Whiteside on grapefruit trees in an organic orchard using an aqueous organic mixture of composted cornmeal, humic acid, molasses, and fish oil vs vegetable oil

Science.gov (United States)

Greasy spot disease of citrus, caused by the fungus Mycosphaerella citri Whiteside, afflicts citrus trees in all citrus-growing areas of the United States, eastern Mexico, Central America, and the Caribbean islands, causing premature defoliation, blemished fruit, and reduced tree vigor, yield, and f...

Sulfur volatiles in guava (Psidium guajava L.) leaves: possible defense mechanism.

Science.gov (United States)

Rouseff, Russell L; Onagbola, Ebenezer O; Smoot, John M; Stelinski, Lukasz L

2008-10-08

Volatiles from crushed and intact guava leaves (Psidium guajava L.) were collected using static headspace SPME and determined using GC-PFPD, pulsed flame photometric detection, and GC-MS. Leaf volatiles from four common citrus culitvars were examined similarly to determine the potential component(s) responsible for guava's protective effect against the Asian citrus psyllid (Diaphorina citri Kuwayama), which is the insect vector of Huanglongbing (HLB) or citrus greening disease. Seven sulfur volatiles were detected: hydrogen sulfide, sulfur dioxide, methanethiol, dimethyl sulfide (DMS), dimethyl disulfide (DMDS), methional, and dimethyl trisulfide (DMTS). Identifications were based on matching linear retention index values on ZB-5, DB-Wax, and PLOT columns and MS spectra in the case of DMDS and DMS. DMDS is an insect toxic, defensive volatile produced only by wounded guava but not citrus leaves and, thus, may be the component responsible for the protective effect of guava against the HLB vector. DMDS is formed immediately after crushing, becoming the major headspace volatile within 10 min. Forty-seven additional leaf volatiles were identified from LRI and MS data in the crushed guava leaf headspace.

Putative sex pheromone of the Asian citrus psyllid, Diaphorina citri, breaks down into an attractant.

Science.gov (United States)

Zanardi, Odimar Z; Volpe, Haroldo X L; Favaris, Arodi P; Silva, Weliton D; Luvizotto, Rejane A G; Magnani, Rodrigo F; Esperança, Victoria; Delfino, Jennifer Y; de Freitas, Renato; Miranda, Marcelo P; Parra, José Roberto P; Bento, José Mauricio S; Leal, Walter S

2018-01-11

Under laboratory conditions, mating activity in Asian citrus psyllid (ACP) started 4 days after emergence, peaked at day 7, and showed a clear window of activity starting 8 h into the photophase and extending through the first hour of the scotophase. We confirmed that ACP males are attracted to emanations from conspecific females. Traps loaded with a candidate compound enriched with female extract, lignoceryl acetate (24Ac), at various doses were active only after being deployed for several weeks in the field, suggesting that a degradation product, not the test compound, was the active ingredient(s). Lignocerol, a possible product of 24Ac degradation, was not active, whereas acetic acid, another possible degradation product, was found in the airborne volatile collections from lures matured under field conditions and detected in higher amounts in volatiles collected from females at the peak of mating activity than in male samples. Acetic acid elicited dose-dependent electroantennographic responses and attracted ACP males, but not females, in Y-type and 4-way olfactometers. Field tests showed that acetic acid-baited traps captured significantly more males than control traps. Surprisingly, captures of females in acetic acid-baited traps were also higher than in control traps, possibly because of physical stimuli emitted by captured males.

H NMR analyses of Citrus macrophylla subjected to Asian citrus psyllid (Diaphorina citri Kuwayama) feeding

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The Asian citrus psyllid (ACP) is a phloem feeding insect that can host and transmit the bacterium Candidatus Liberibacter asiaticus (CLas), which is the putative causative agent of the economically important citrus disease, Huanglongbing (HLB). ACP are widespread in Florida, and are spreading in Ca...

Structural features of PhoX, one of the phosphate-binding proteins from Pho regulon of Xanthomonas citri

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Pegos, Vanessa R.; Santos, Rodrigo M. L.; Medrano, Francisco J.

2017-01-01

In Escherichia coli, the ATP-Binding Cassette transporter for phosphate is encoded by the pstSCAB operon. PstS is the periplasmic component responsible for affinity and specificity of the system and has also been related to a regulatory role and chemotaxis during depletion of phosphate. Xanthomonas citri has two phosphate-binding proteins: PstS and PhoX, which are differentially expressed under phosphate limitation. In this work, we focused on PhoX characterization and comparison with PstS. The PhoX three-dimensional structure was solved in a closed conformation with a phosphate engulfed in the binding site pocket between two domains. Comparison between PhoX and PstS revealed that they originated from gene duplication, but despite their similarities they show significant differences in the region that interacts with the permeases. PMID:28542513

Lethal and Sublethal Effects of Insecticides Used on Citrus, on the Ectoparasitoid Tamarixia radiata.

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Beloti, Vitor Hugo; Alves, Gustavo Rodrigues; Araújo, Diogo Feliciano Dias; Picoli, Mateus Manara; Moral, Rafael de Andrade; Demétrio, Clarice Garcia Borges; Yamamoto, Pedro Takao

2015-01-01

Huanglongbing (HLB) is a disease associated with the bacteria "Candidatus Liberibacter spp." and has been devastating citrus orchards around the world. Its management involves control of the insect vector, the Asian citrus psyllid Diaphorina citri Kuwayama. However, the indiscriminate use of chemicals has caused pest outbreaks and eliminated the natural enemies of the vector, such as the parasitoid Tamarixia radiata (Waterston), the main agent for biological control of D. citri. This study assessed the lethal and sublethal effects of insecticides recommended for integrated production of citrus on the parasitoid T. radiata. When adult parasitoids were exposed to residues of 25 insecticides, 20% of them, i.e., gamma-cyhalothrin, etofenprox, azadirachtin, tebufenozide and pyriproxyfen, were considered as harmless (Class 1), 12% as slightly harmful (Class 2), 12% as moderately harmful (Class 3) and 56% as harmful (Class 4), according to the classification proposed by the IOBC/WPRS. Afterward, 14 insecticides (5 harmless and 9 harmful) were sprayed on the parasitoid pupae. Of the 14 insecticides tested, only the organophosphates dimethoate and chlorpyrifos affected the parasitoid emergence. The effects of insecticides on the parasitism capacity of adults exposed to residues of azadirachtin, etofenprox, gamma-cyhalothrin, pyriproxyfen and tebufenozide (harmless) were also evaluated. Tebufenozide and gamma-cyhalothrin affected the parasitism of the F0 generation, but did not affect the emergence of the F1 and F2 generations. Therefore, for an effective IPM program, selective insecticides or harmful pesticides to adult parasitoids could be used in the field, provided that the adults do not occur naturally and the chemical applications do not coincide with parasitoid releases.

Lethal and Sublethal Effects of Insecticides Used on Citrus, on the Ectoparasitoid Tamarixia radiata.

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Vitor Hugo Beloti

Full Text Available Huanglongbing (HLB is a disease associated with the bacteria "Candidatus Liberibacter spp." and has been devastating citrus orchards around the world. Its management involves control of the insect vector, the Asian citrus psyllid Diaphorina citri Kuwayama. However, the indiscriminate use of chemicals has caused pest outbreaks and eliminated the natural enemies of the vector, such as the parasitoid Tamarixia radiata (Waterston, the main agent for biological control of D. citri. This study assessed the lethal and sublethal effects of insecticides recommended for integrated production of citrus on the parasitoid T. radiata. When adult parasitoids were exposed to residues of 25 insecticides, 20% of them, i.e., gamma-cyhalothrin, etofenprox, azadirachtin, tebufenozide and pyriproxyfen, were considered as harmless (Class 1, 12% as slightly harmful (Class 2, 12% as moderately harmful (Class 3 and 56% as harmful (Class 4, according to the classification proposed by the IOBC/WPRS. Afterward, 14 insecticides (5 harmless and 9 harmful were sprayed on the parasitoid pupae. Of the 14 insecticides tested, only the organophosphates dimethoate and chlorpyrifos affected the parasitoid emergence. The effects of insecticides on the parasitism capacity of adults exposed to residues of azadirachtin, etofenprox, gamma-cyhalothrin, pyriproxyfen and tebufenozide (harmless were also evaluated. Tebufenozide and gamma-cyhalothrin affected the parasitism of the F0 generation, but did not affect the emergence of the F1 and F2 generations. Therefore, for an effective IPM program, selective insecticides or harmful pesticides to adult parasitoids could be used in the field, provided that the adults do not occur naturally and the chemical applications do not coincide with parasitoid releases.

Formation of Stylet Sheaths in āere (in air from eight species of phytophagous hemipterans from six families (Suborders: Auchenorrhyncha and Sternorrhyncha.

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J Kent Morgan

Full Text Available Stylet sheath formation is a common feature among phytophagous hemipterans. These sheaths are considered essential to promote a successful feeding event. Stylet sheath compositions are largely unknown and their mode of solidification remains to be elucidated. This report demonstrates the formation and solidification of in āere (in air produced stylet sheaths by six hemipteran families: Diaphorina citri (Psyllidae, Asian citrus psyllid, Aphis nerii (Aphididae, oleander/milkweed aphid, Toxoptera citricida (Aphididae, brown citrus aphid, Aphis gossypii (Aphididae, cotton melon aphid, Bemisia tabaci biotype B (Aleyrodidae, whitefly, Homalodisca vitripennis (Cicadellidae, glassy-winged sharpshooter, Ferrisia virgata (Pseudococcidae, striped mealybug, and Protopulvinaria pyriformis (Coccidae, pyriform scale. Examination of in āere produced stylet sheaths by confocal and scanning electron microscopy shows a common morphology of an initial flange laid down on the surface of the membrane followed by continuous hollow core structures with sequentially stacked hardened bulbous droplets. Single and multi-branched sheaths were common, whereas mealybug and scale insects typically produced multi-branched sheaths. Micrographs of the in āere formed flanges indicate flange sealing upon stylet bundle extraction in D. citri and the aphids, while the B. tabaci whitefly and H. vitripennis glassy-winged sharpshooter flanges remain unsealed. Structural similarity of in āere sheaths are apparent in stylet sheaths formed in planta, in artificial diets, or in water. The use of 'Solvy', a dissolvable membrane, for intact stylet sheath isolation is reported. These observations illustrate for the first time this mode of stylet sheath synthesis adding to the understanding of stylet sheath formation in phytophagous hemipterans and providing tools for future use in structural and compositional analysis.

Contrasting amino acid profiles among permissive and non-permissive hosts of Candidatus Liberibacter asiaticus, putative causal agent of Huanglongbing.

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Mamoudou Sétamou

Full Text Available Huanglongbing is a devastating disease of citrus. In this study, a comprehensive profile of phloem sap amino acids (AA in four permissive host plants of Candidatus Liberibacter asiaticus (CLas and three non-permissive Rutaceae plants was conducted to gain a better understanding of host factors that may promote or suppress the bacterium. The AA profiles of Diaphorina citri nymphs and adults were similarly analyzed. A total of 38 unique AAs were detected in phloem sap of the various plants and D. citri samples, with phloem sap of young shoots containing more AAs and at higher concentrations than their mature counterparts. All AAs detected in phloem sap of non-permissive plants were also present in CLas -permissive hosts plus additional AAs in the latter class of plants. However, the relative composition of 18 commonly shared AAs varied between CLas -permissive hosts and non-permissive plants. Multivariate analysis with a partial least square discriminant methodology revealed a total of 12 AAs as major factors affecting CLas host status, of which seven were positively related to CLas tolerance/resistance and five positively associated with CLas susceptibility. Most of the AAs positively associated with CLas susceptibility were predominantly of the glutamate family, notably stressed-induced AAs such as arginine, GABA and proline. In contrast, AAs positively correlated with CLas tolerance/resistance were mainly of the serine family. Further analysis revealed that whereas the relative proportions of AAs positively associated with CLas susceptibility did not vary with host developmental stages, those associated with CLas tolerance/resistance increased with flush shoot maturity. Significantly, the proline-to-glycine ratio was determined to be an important discriminating factor for CLas permissivity with higher values characteristic of CLas -permissive hosts. This ratio could be exploited as a biomarker in HLB-resistance breeding programs.

Nucleotides, micro- and macro-nutrients, limonoids, flavonoids, and hydroxycinnamates composition in the phloem sap of sweet orange.

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Hijaz, Faraj; Manthey, John A; Van der Merwe, Deon; Killiny, Nabil

2016-06-02

Currently, the global citrus production is declining due to the spread of Huanglongbing (HLB). HLB, otherwise known as citrus greening, is caused by Candidatus Liberibacter asiaticus (CLas) and is transmitted by the Asian citrus psyllids (ACP), Diaphorina citri Kuwayama. ACP transmits CLas bacterium while feeding on the citrus phloem sap. Multiplication of CLas in the phloem of citrus indicates that the sap contains all the essential nutrients needed for CLas. In this study, we investigated the micro- and macro-nutrients, nucleotides, and others secondary metabolites of phloem sap from pineapple sweet orange. The micro- and macro-nutrients were analyzed using inductively coupled plasma-mass spectroscopy (ICP-MS) and inductively coupled plasma-optical emission spectroscopy (ICP-OES). Nucleotides and other secondary metabolites analysis was accomplished by reversed phase HPLC coupled with UV, fluorescence detection, or negative mode electrospray ionization mass spectrometry (ESI-MS). Calcium (89 mM) was the highest element followed by potassium (38.8 mM) and phosphorous (24 mM). Magnesium and sulfur were also abundant and their concentrations were 15 and 9 mM, respectively. The rest of the elements were found in low amounts (sap.

Comparison of Potato and Asian Citrus Psyllid Adult and Nymph Transcriptomes Identified Vector Transcripts with Potential Involvement in Circulative, Propagative Liberibacter Transmission

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Tonja W. Fisher

2014-11-01

Full Text Available The potato psyllid (PoP Bactericera cockerelli (Sulc and Asian citrus psyllid (ACP Diaphorina citri Kuwayama are the insect vectors of the fastidious plant pathogen, Candidatus Liberibacter solanacearum (CLso and Ca. L. asiaticus (CLas, respectively. CLso causes Zebra chip disease of potato and vein-greening in solanaceous species, whereas, CLas causes citrus greening disease. The reliance on insecticides for vector management to reduce pathogen transmission has increased interest in alternative approaches, including RNA interference to abate expression of genes essential for psyllid-mediated Ca. Liberibacter transmission. To identify genes with significantly altered expression at different life stages and conditions of CLso/CLas infection, cDNA libraries were constructed for CLso-infected and -uninfected PoP adults and nymphal instars. Illumina sequencing produced 199,081,451 reads that were assembled into 82,224 unique transcripts. PoP and the analogous transcripts from ACP adult and nymphs reported elsewhere were annotated, organized into functional gene groups using the Gene Ontology classification system, and analyzed for differential in silico expression. Expression profiles revealed vector life stage differences and differential gene expression associated with Liberibacter infection of the psyllid host, including invasion, immune system modulation, nutrition, and development.

Mass distribution and spatial organization of the linear bacterial motor of Spiroplasma citri R8A2.

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Trachtenberg, Shlomo; Andrews, S Brian; Leapman, Richard D

2003-03-01

In the simple, helical, wall-less bacterial genus Spiroplasma, chemotaxis and motility are effected by a linear, contractile motor arranged as a flat cytoskeletal ribbon attached to the inner side of the membrane along the shortest helical line. With scanning transmission electron microscopy and diffraction analysis, we determined the hierarchical and spatial organization of the cytoskeleton of Spiroplasma citri R8A2. The structural unit appears to be a fibril, approximately 5 nm wide, composed of dimers of a 59-kDa protein; each ribbon is assembled from seven fibril pairs. The functional unit of the intact ribbon is a pair of aligned fibrils, along which pairs of dimers form tetrameric ring-like repeats. On average, isolated and purified ribbons contain 14 fibrils or seven well-aligned fibril pairs, which are the same structures observed in the intact cell. Scanning transmission electron microscopy mass analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified cytoskeletons indicate that the 59-kDa protein is the only constituent of the ribbons.

The GC/MS Analysis of Volatile Components Extracted by Different Methods from Exocarpium Citri Grandis

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Zhisheng Xie

2013-01-01

Full Text Available Volatile components from Exocarpium Citri Grandis (ECG were, respectively, extracted by three methods, that is, steam distillation (SD, headspace solid-phase microextraction (HS-SPME, and solvent extraction (SE. A total of 81 compounds were identified by gas chromatography-mass spectrometry including 77 (SD, 56 (HS-SPME, and 48 (SE compounds, respectively. Despite of the extraction method, terpenes (39.98~57.81% were the main volatile components of ECG, mainly germacrene-D, limonene, 2,6,8,10,14-hexadecapentaene, 2,6,11,15-tetramethyl-, (E,E,E-, and trans-caryophyllene. Comparison was made among the three methods in terms of extraction profile and property. SD relatively gave an entire profile of volatile in ECG by long-time extraction; SE enabled the analysis of low volatility and high molecular weight compounds but lost some volatiles components; HS-SPME generated satisfactory extraction efficiency and gave similar results to those of SD at analytical level when consuming less sample amount, shorter extraction time, and simpler procedure. Although SD and SE were treated as traditionally preparative extractive techniques for volatiles in both small batches and large scale, HS-SPME coupled with GC/MS could be useful and appropriative for the rapid extraction and qualitative analysis of volatile components from medicinal plants at analytical level.

Expression, purification, crystallization and preliminary X-ray analysis of YaeQ (XAC2396) from Xanthomonas axonopodis pv. citri

International Nuclear Information System (INIS)

Guzzo, Cristiane R.; Nagem, Ronaldo A. P.; Galvão-Botton, Leonor M. P.; Guimarães, Beatriz G.; Medrano, Francisco J.; Barbosa, João A. R. G.; Farah, Chuck S.

2005-01-01

The first crystallographic study of a member of the YaeQ family of proteins, which are conserved in a small group of Gram-negative bacteria, most of which are animal or plant pathogens, is reported. Diffraction data were collected to 1.9 Å resolution and an interpretable electron-density map was obtained. Xanthomonas axonopodis pv. citri YaeQ (XAC2396) is a member of a family of bacterial proteins conserved in several Gram-negative pathogens. Here, the cloning, expression, purification and crystallization of the 182-residue (20.6 kDa) YaeQ protein are described. Recombinant YaeQ containing selenomethionine was crystallized in space group P2 1 and crystals diffracted to 1.9 Å resolution at a synchrotron source. The unit-cell parameters are a = 39.75, b = 91.88, c = 48.03 Å, β = 108.37°. The calculated Matthews coefficient suggests the presence of two YaeQ molecules in the asymmetric unit. Initial experimental phases were calculated by the multiple-wavelength anomalous dispersion technique and an interpretable electron-density map was obtained

Mosquitocidal carbamates with low toxicity to agricultural pests: an advantageous property for insecticide resistance management.

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Swale, Daniel R; Carlier, Paul R; Hartsel, Joshua A; Ma, Ming; Bloomquist, Jeffrey R

2015-08-01

Insecticide resistance in the malaria mosquito Anopheles gambiae is well documented, and widespread agricultural use of pyrethroids may exacerbate development of resistance when pyrethroids are used in vector control. We have developed carbamate anticholinesterases that possess a high degree of An. gambiae:human selectivity for enzyme inhibition. The purpose of this study was to assess the spectrum of activity of these carbamates against other mosquitoes and agricultural pests. Experimental carbamates were potent inhibitors of mosquito acetylcholinesterases, with IC50 values in the nanomolar range. Similar potencies were observed for Musca domestica and Drosophila melanogaster enzymes. Although meta-substituted carbamates were potent inhibitors, two ortho-substituted carbamates displayed poor enzyme inhibition (IC50 ≥ 10(-6)  M) in honey bee (Apis mellifera), Asian citrus psyllid (Diaphorina citri) and lepidopteran agricultural pests (Plutella xylostella and Ostrinia nubilalis). Enzyme inhibition results were confirmed by toxicity studies in caterpillars, where the new carbamates were 2-3-fold less toxic than propoxur and up to tenfold less active than bendiocarb, indicating little utility of these compounds for crop protection. The experimental carbamates were broadly active against mosquito species but not agricultural pests, which should mitigate selection for mosquito insecticide resistance by reducing agricultural uses of these compounds. © 2014 Society of Chemical Industry. © 2014 Society of Chemical Industry.

Metalized polyethylene mulch to repel Asian citrus psyllid, slow spread of huanglongbing and improve growth of new citrus plantings.

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Croxton, Scott D; Stansly, Philip A

2014-02-01

Greening or huanglongbing (HLB) is a debilitating disease of citrus caused by Candidatus Liberibactor asiaticus and transmitted by the Asian citrus psyllid (ACP), Diaphorina citri. HLB now occurs worldwide in all major citrus growing regions except the Mediterranean and Australia. Management relies principally on insecticidal control of the ACP vector, but is insufficient, even for young trees which are most susceptible to the disease. We tested the ability of metalized polyethylene mulch to repel adult ACP as well as effects on incidence of HLB and early tree growth. Metalized mulch significantly reduced ACP populations and HLB incidence compared to whiteface mulch or bare ground. In addition, metalized mulch, together with the associated drip irrigation and fertigation system, increased soil moisture, reduced weed pressure, and increased tree growth rate. Metalized mulch slows spread of ACP and therefore HLB pressure on young citrus trees. Metalized mulch can thereby augment current control measures for young trees based primarily on systemic insecticides. Additional costs could be compensated for by increased tree growth rate which would shorten time to crop profitability. These advantages make a compelling case for large-scale trials using metalized mulch in young citrus plantings threatened by HLB. © 2013 Society of Chemical Industry.

Semiochemical and Vibrational Cues and Signals Mediating Mate Finding and Courtship in Psylloidea (Hemiptera: A Synthesis

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Umar K. Lubanga

2014-07-01

Full Text Available Mate finding and courtship involve complex interactions that require close coordination between individuals of the opposite gender. Well-organized signalling systems, sometimes involving a combination of signal modalities, are required to convey species-specific and individual information to members of the opposite gender. Previous studies of psyllids have focused on single-signal modalities and have largely ignored the potentially interdependent nature of different types of signals. Several studies have shown that semiochemicals play a role in psyllid mate finding. However, long-range semiochemical sex attractants, such as the highly volatile sex pheromones used by many Lepidoptera (molecular weights <300, are yet to be identified. The compounds identified thus far, namely 13-methylheptacosane (from Cacopsylla pyricola and dodecanoic acid (from Diaphorina citri, seem to have short range activity or no activity under field conditions. The possible role played by cuticular hydrocarbons in psyllid courtship remains largely ignored. Conversely, many psyllid species rely on vibrational signals for mate finding and mate assessment during courtship. This apparent disproportional reliance on vibrational rather than semiochemical signals suggests that vibrational signals have been more influential in sexual selection in psyllids. However, male fitness, female choice and benefits accrued from selecting fitter males remain poorly understood.

Changes in Variable Number of Tandem Repeats in 'Candidatus Liberibacter asiaticus' through Insect Transmission.

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Hiroshi Katoh

Full Text Available Citrus greening (huanglongbing is the most destructive citrus disease worldwide. The disease is associated with three species of 'Candidatus Liberibacter' among which 'Ca. Liberibacter asiaticus' has the widest distribution. 'Ca. L. asiaticus' is commonly transmitted by a phloem-feeding insect vector, the Asian citrus psyllid Diaphorina citri. A previous study showed that isolates of 'Ca. L. asiaticus' were clearly differentiated by variable number of tandem repeat (VNTR profiles at four loci in the genome. In this study, the VNTR analysis was further validated by assessing the stability of these repeats after multiplication of the pathogen upon host-to-host transmission using a 'Ca. L. asiaticus' strain from Japan. The results showed that some tandem repeats showed detectable changes after insect transmission. To our knowledge, this is the first report to demonstrate that the repeat numbers VNTR 002 and 077 of 'Ca. L. asiaticus' change through psyllid transmission. VNTRs in the recipient plant were apparently unrelated to the growing phase of the vector. In contrast, changes in the number of tandem repeats increased with longer acquisition and inoculation access periods, whereas changes were not observed through psyllid transmission after relatively short acquisition and inoculation access periods, up to 20 and 19 days, respectively.

Costs and benefits of insecticide and foliar nutrient applications to huanglongbing-infected citrus trees.

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Tansey, James A; Vanaclocha, Pilar; Monzo, Cesar; Jones, Moneen; Stansly, Philip A

2017-05-01

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae), vectors Candidatus Liberibacter asiaticus, which causes huanglongbing (HLB). In Florida, HLB incidence is approaching 100% statewide. Yields have decreased and production costs have increased since 2005. Despite this, some growers are maintaining a level of production and attribute this in part to aggressive psyllid control and foliar nutrition sprays. However, the value of these practices is debated. A replicated field study was initiated in 2008 in a commercial block of 'Valencia' sweet orange trees to evaluate individual and combined effects of foliar nutrition and ACP control. Results from 2012-2016 are presented. Insecticides consistently reduced ACP populations. However, neither insecticide nor nutrition applications significantly influenced HLB incidence or PCR copy number in mature trees. In reset trees, infection continued to build and reached 100% in all treatments. Greatest yields (kg fruit ha -1 ) and production (kg solids ha -1 ) were obtained from trees receiving both insecticides and foliar nutrition. All treatments resulted in production and financial gains relative to controls. However, material and application costs associated with the nutrition component offset these gains, resulting in lesser benefits than insecticides applied alone. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Evaluation of systemic neonicotinoid insecticides for the management of the Asian citrus psyllid Diaphorina citri on containerized citrus.

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Byrne, Frank J; Daugherty, Matthew P; Grafton-Cardwell, Elizabeth E; Bethke, James A; Morse, Joseph G

2017-03-01

Studies were conducted to evaluate uptake and retention of three systemic neonicotinoid insecticides, dinotefuran, imidacloprid and thiamethoxam, in potted citrus nursery plants treated at standard label rates. Infestation of these plants placed at a field site with moderate levels of Asian citrus psyllid (ACP) was monitored for 14 weeks following treatments, and insecticide residues in leaf tissue were quantified using enzyme-linked immunosorbent assay (ELISA). Bioassays were conducted using leaves harvested on various dates post-treatment to compare the efficacies of residues against adult ACP. Residues of the three neonicotinoids were detected in leaf tissues within 1 week after treatment. Peak concentrations established at 1 week for imidacloprid and dinotefuran and at 2 weeks for thiamethoxam. Imidacloprid and thiamethoxam outperformed the control and dinotefuran treatments at protecting trees from infestations by ACP eggs and nymphs. For a given insecticide concentration in leaf tissue, thiamethoxam induced the highest mortality of the three insecticides, and dinotefuran was the least toxic. If the time needed to achieve effective thresholds of a systemic neonicotinoid is known, treatments at production facilities could be scheduled that would minimize unnecessary post-treatment holding periods and ensure maximum retention of effective concentrations after the plants have shipped to retail outlets. The rapid uptake of the insecticides and retention at effective concentrations in containerized citrus suggest that the current 30 day post-treatment shipping restriction from production facilities to retail outlets outside of quarantine could be shortened to 14 days. Thiamethoxam should be added to the list of approved nursery treatments. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Multi-criteria optimization for ultrasonic-assisted extraction of antioxidants from Pericarpium Citri Reticulatae using response surface methodology, an activity-based approach.

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Zeng, Shanshan; Wang, Lu; Zhang, Lei; Qu, Haibin; Gong, Xingchu

2013-06-01

An activity-based approach to optimize the ultrasonic-assisted extraction of antioxidants from Pericarpium Citri Reticulatae (Chenpi in Chinese) was developed. Response surface optimization based on a quantitative composition-activity relationship model showed the relationships among product chemical composition, antioxidant activity of extract, and parameters of extraction process. Three parameters of ultrasonic-assisted extraction, including the ethanol/water ratio, Chenpi amount, and alkaline amount, were investigated to give optimum extraction conditions for antioxidants of Chenpi: ethanol/water 70:30 v/v, Chenpi amount of 10 g, and alkaline amount of 28 mg. The experimental antioxidant yield under the optimum conditions was found to be 196.5 mg/g Chenpi, and the antioxidant activity was 2023.8 μmol Trolox equivalents/g of the Chenpi powder. The results agreed well with the second-order polynomial regression model. This presented approach promised great application potentials in both food and pharmaceutical industries. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Disease Development and Symptom Expression of Xanthomonas axonopodis pv. citri in Various Citrus Plant Tissues.

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Vernière, C J; Gottwald, T R; Pruvost, O

2003-07-01

ABSTRACT Experimental inoculations of Xanthomonas axonopodis pv. citri in different tissues of Tahiti lime and Pineapple sweet orange were conducted monthly under natural conditions on Réunion Island. The interactions between a set of environmental and epidemic variables associated with disease expression and 184 different factor combinations were investigated to determine the parameters needed to explain Asiatic citrus canker (ACC) disease expression. Area under the disease progress curve (AUDPC), inoculation date (Id), fruit and leaf age ratings (FAR and LAR), and number of days during the first 2 weeks postinoculation for which the temperature was less than 14 degrees C (T(min)) or more than 28 degrees C (T(max)) were retained by principal component analysis and canonical correlation analysis as the most meaningful epidemic and environmental variables, respectively. AUDPC as the strongest dependent variable and combinations of the environmental variables as independent variables were used in multiple regression analyses. Tissue age rating at the time of infection was a good predictor for disease resulting from spray inoculation on fruits and leaves and also on fruits following a wound inoculation. Temperature, as expressed by T(min) or T(max), was also a significant factor in determining disease development described by AUDPC. Mature green stems were highly susceptible after wounding, similarly to leaves, but buds and leaf scars expressed the lowest susceptibility. These variations in disease expression according to the tissues will have different impacts on ACC epidemiology.

Nucleotides, micro- and macro-nutrients, limonoids, flavonoids, and hydroxycinnamates composition in the phloem sap of sweet orange

Science.gov (United States)

Hijaz, Faraj; Manthey, John A.; Van der Merwe, Deon; Killiny, Nabil

2016-01-01

ABSTRACT Currently, the global citrus production is declining due to the spread of Huanglongbing (HLB). HLB, otherwise known as citrus greening, is caused by Candidatus Liberibacter asiaticus (CLas) and is transmitted by the Asian citrus psyllids (ACP), Diaphorina citri Kuwayama. ACP transmits CLas bacterium while feeding on the citrus phloem sap. Multiplication of CLas in the phloem of citrus indicates that the sap contains all the essential nutrients needed for CLas. In this study, we investigated the micro- and macro-nutrients, nucleotides, and others secondary metabolites of phloem sap from pineapple sweet orange. The micro- and macro-nutrients were analyzed using inductively coupled plasma-mass spectroscopy (ICP-MS) and inductively coupled plasma-optical emission spectroscopy (ICP-OES). Nucleotides and other secondary metabolites analysis was accomplished by reversed phase HPLC coupled with UV, fluorescence detection, or negative mode electrospray ionization mass spectrometry (ESI-MS). Calcium (89 mM) was the highest element followed by potassium (38.8 mM) and phosphorous (24 mM). Magnesium and sulfur were also abundant and their concentrations were 15 and 9 mM, respectively. The rest of the elements were found in low amounts (flavonoids. In addition, several hydroxycinnamates were detected. The results of this study will increase our knowledge about the nature and the chemical composition of citrus phloem sap. PMID:27171979

PHLOEM PROMOTERS IN TRANSGENIC SWEET ORANGE ARE DIFFERENTIALLY TRIGGERED BY Candidatus Liberibacter asiaticus

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LUZIA YURIKO MIYATA

Full Text Available ABSTRACT The use of promoters preferentially expressed in specific plant tissues is a desirable strategy to search for resistance for pathogens that colonize these tissues. The bacterium Candidatus Liberibacter asiaticus (Las, associated with huanglongbing disease (HLB of citrus, colonizes phloem vessels. Some promoters, besides conferring tissue-specific expression, can also respond to the presence of the pathogen. The objective of the present study was to verify if the presence of Las could modulate the activation of the phloem-specific promoters AtPP2 (Arabidopsis thaliana phloem protein 2, AtSUC2 (A. thaliana sucrose transporter 2 and CsPP2 ( pCitrus phloemrotein 2, known to be expressed in Citrus sinensis phloem. ‘Hamlin’ sweet orange plants (Citrus sinensis L. Osbeck transformed with the uidA (GUS reporter gene under the control of AtPP2, AtSUC2 and CsPP2 promoters were infected to evaluate the interdependence between transgene expression and the concentration of Las. Plants were inoculated with Las by Diaphorina citri and eighteen months later, bacterial concentration and uidA expression were determined by qPCR and RT-qPCR, respectively. Reporter gene expression driven by AtSUC2 promoter was strongly and positively correlated with Las concentration. Therefore, this promoter combines desirable features of both tissue-specificity and pathogen-inducibility for the production of transgenic plants tolerant to Las.

Fortuitous establishment of Ageniaspis citricola (Hymenoptera: Encyrtidae) in Jamaica on the citrus leafminer (Lepidoptera: Gracillariidae)

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Hoy, M.A.; Jeyaprakash, A.; Clarke-Harris, D.

2007-01-01

These data indicate that the population of A. citricola in Jamaica probably originated from the Australian (Thailand), rather than from the Taiwan, population. This is consistent with what is currently known about the origin of the established Ageniaspis population in Florida (Alvarez 2000). It is not known when, or how, A. citricola arrived in Jamaica, although the CLM was detected there in 1994. The fortuitous establishment of A. citricolaon the CLM in Jamaica is not the only such establishment of a natural enemy discovered during this 2004 survey of citrus. The parasitoid Lipolexis oregmae Gahan (Hymenoptera: Aphidiidae) was found attacking the brown citrus aphid, Toxoptera citricida Kirkaldy (Hemiptera: Aphididae) (Hoy et al., unpublished data), and the eulophid parasitoid Tamarixia radiata Waterston was found attacking the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae). The fact that 3 natural enemies of 3 invasive citrus pests were found in Jamaica, none of which were purposefully imported and released, suggests that pest-infested citrus trees were imported into Jamaica without going through appropriate quarantine procedures. Because each pest arrived at different times, the parasitoids probably arrived at different times, as well. This indicates that an analysis is needed to identify the critical control points within those services in Jamaica that support border protection, and that procedures may require strengthening. (author)

Amplification of DNA of Xanthomonas axonopodis pv. citri from historic citrus canker herbarium specimens.

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Li, Wenbin; Brlansky, Ronald H; Hartung, John S

2006-05-01

Herbaria are important resources for the study of the origins and dispersal of plant pathogens, particularly bacterial plant pathogens that incite local lesions in which large numbers of pathogen genomes are concentrated. Xanthomonas axonopodis pv. citri (Xac), the causal agent of citrus bacterial canker disease, is a notable example of such a pathogen. The appearance of novel strains of the pathogen in Florida and elsewhere make it increasingly important to understand the relationships among strains of this pathogen. USDA-ARS at Beltsville, Maryland maintains approximately 700 herbarium specimens with citrus canker disease lesions up to 90 years old, originally collected from all over the world, and so is an important resource for phytogeographic studies of this bacterium. Unfortunately, DNA in herbarium specimens is degraded and may contain high levels of inhibitors of PCR. In this study, we compared a total of 23 DNA isolation techniques in combination with 31 novel primer pairs in order to develop an efficient protocol for the analysis of Xac DNA in herbarium specimens. We identified the most reliable extraction method, identified in terms of successful amplification by our panel of 31 primer pairs. We also identified the most robust primer pairs, identified as successful in the largest number of extracts prepared by different methods. We amplified Xac genomic sequences up to 542 bp long from herbarium samples up to 89 years old. Primers varied in effectiveness, with some primer pairs amplifying Xac DNA from a 1/10,000 dilution of extract from a single lesion from a citrus canker herbarium specimen. Our methodology will be useful to identify pathogens and perform molecular analyses of bacterial and possibly fungal genomes from herbarium specimens.

The weathervane model, a functional and structural organization of the two-component alkanesulfonate oxidoreductase SsuD from Xanthomonas citri

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Pegos, V.R.; Oliveira, P.S.L.; Balan, A.

2012-01-01

Full text: In Xanthomonas citri, the phytopathogen responsible for the canker citrus disease, we identified in the ssuABCDE operon, genes encoding the alkanesulfonate ABC transporter as well as the two enzymes responsible for oxido reduction of the respective substrates. SsuD and SsuE proteins represent a two-component system that can be assigned to the group of FMNH 2 -dependent monooxygenases. How- ever, despite of the biochemical information about SsuD and SsuE orthologs from Escherichia coli, there is no structural information of how the two proteins work together. In this work, we used ultracentrifugation, SAXS data and molecular modeling to construct a structural/functional model, which consists of eight molecules organized in a weathervane shape. Through this model, SsuD ligand-binding site for NADPH 2 and FMN substrates is clearly exposed, in a way that might allow the protein-protein interactions with SsuE. Moreover, based on molecular dynamics simulations of SsuD in apo state, docked with NADPH 2 , FMN or both substrates, we characterized the residues of the pocket, the mechanism of substrate interaction and transfer of electrons from NADPH 2 to FMN. This is the first report that links functional and biochemical data with structural analyses. (author)

Compatibility and efficacy of Isaria fumosorosea with horticultural oils for mitigation of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae)

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Horticultural oils are an important component of integrated management programs of several phytophagous arthropods, and pathogens affecting fruit, ornamental and vegetables in greenhouse and field production systems. Although effective against the target pest, their incompatibility with biological c...

Efficacy of an autodisseminator of an entomopathogenic fungus, Isaria fumosorosea, to suppress Asian citrus psyllid, Diaphorina citri, under greenhouse conditions

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The Asian citrus psyllid (ACP), transmits the causative agents of citrus greening disease or huanglongbing (HLB), the most devastating disease of citrus trees in the world today. ACP dwelling in noncommercial citrus (neighborhoods, commercial landscapes, etc.) can stymie area-wide management program...

EFFECTIVENESS OF HUANGLONGBING VECTOR (DIAPHORINA CITRI KUWAYAMA CONTROL IN CITRUS GROWER GROUP BASED IN SAMBAS REGENCY OF WEST KALIMANTAN, INDONESIA

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Supriyanto A.

2017-12-01

Full Text Available The purpose of this study was to determine the effectiveness of Huanglongbing vector control based on Citrus Grower Group recommendation. Studies have been carried out in 2010 in Tebas Sungai village, Sambas district, with 11 tangerine groves owned by growers in the Citrus grower Association of Sambas district. The tangerine grove that been used are, one grower's orchard as a demonstration plot in a particular citrus grower group (orchard I; five other citrus orchards with different ownership at the same citrus grower Group (orchard II, as well as five other citrus orchard with different ownership which each of them spreads over five different citrus grower groups outside the farm demonstration plots (orchard III. The recommendation technology for controlling Huanglongbing vector which applied in this experiment, included bark painting by systemic insecticide of imidacloprid for two each 1.5-month and spray using contact insecticide with dimethoate to the plant crown which application time been alternated after bark painting application. The effectiveness of technology implementation is measured by a decrease psyllid populations found in citrus samples in adult stage, nymphs and eggs that were observed at regular intervals every two weeks during the flushing to the 14th week after the first treatment. The results showed that recommended treatment technology were absolutely proven to reduce Huanglongbing vector population in significant, namely in the orchard I, II, and III respectively at 95.3%, 84.7%, and 72% for stage adult; 97.3 %, 80%, and 100% for stage nymphs; and 98.5%, 100% and 100% for the egg stage.

Silencing abnormal wing disc gene of the Asian citrus psyllid, Diaphorina citri disrupts adult wing development and increases nymph mortality.

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Ibrahim El-Shesheny

Full Text Available Huanglongbing (HLB causes considerable economic losses to citrus industries worldwide. Its management depends on controlling of the Asian citrus Psyllid (ACP, the vector of the bacterium, Candidatus Liberibacter asiaticus (CLas, the causal agent of HLB. Silencing genes by RNA interference (RNAi is a promising tool to explore gene functions as well as control pests. In the current study, abnormal wing disc (awd gene associated with wing development in insects is used to interfere with the flight of psyllids. Our study showed that transcription of awd is development-dependent and the highest level was found in the last instar (5(th of the nymphal stage. Micro-application (topical application of dsRNA to 5(th instar of nymphs caused significant nymphal mortality and adult wing-malformation. These adverse effects in ACP were positively correlated with the amounts of dsRNA used. A qRT-PCR analysis confirmed the dsRNA-mediated transcriptional down-regulation of the awd gene. Significant down-regulation was required to induce a wing-malformed phenotype. No effect was found when dsRNA-gfp was used, indicating the specific effect of dsRNA-awd. Our findings suggest a role for awd in ACP wing development and metamorphosis. awd could serve as a potential target for insect management either via direct application of dsRNA or by producing transgenic plants expressing dsRNA-awd. These strategies will help to mitigate HLB by controlling ACP.

3-Nitropropionic acid production by the endophytic Diaporthe citri: Molecular taxonomy, chemical characterization, and quantification under pH variation.

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Polonio, Julio Cesar; Ribeiro, Marcos Alessandro Dos Santos; Rhoden, Sandro Augusto; Sarragiotto, Maria Helena; Azevedo, João Lúcio; Pamphile, João Alencar

2016-12-01

3-nitropropionic acid (3-NPA) is a nitrogenated compound produced by plants and fungi and has been associated with poisoning episodes in humans, animals, and to induction of Huntington disease symptoms in rats. The production of 3-NPA by endophytes has been reported, but the function and biosynthesis are not well-defined. The specie of endophytic strain G-01 was confirmed as Diaporthe citri using a multilocus sequence analysis, and was verified different concentrations of 3-NPA produced at different initial pHs by these strain. The chemical analysis indicated that 3-NPA was the majority compound present in the crude extracts. The better extraction condition was at an initial pH of 7.0 for 22 d, yielding about 80 % of 3-NPA per mg of extract. It was observed that the concentration of 3-NPA increased after the initial consumption of reduction sugars, indicating that the compound is produced after the high energetic production phase of the fungus. These and other studies demonstrate the production of this compound by plants and endophytic fungi, indicating that 3-NPA may be involved in defence and nutrition systems of endophytes and host plants, and they also might participate in the biogeochemical nitrogen cycle. Copyright © 2016 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Immunogenesity of spesific protein molecular weight 16 KDa (PS16 leaf of siam citrus infected by citrus vein phloem degeneration (CVPD disease

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Made Sritamin

2012-02-01

Full Text Available Citrus Vein Phloem degeneration (CVPD is an important citrus disese, which damaged citrus plantation and causing decrease of citrus production. In Indonesia, the CVPD disease caused by Liberobacter asiaticum bactery and the disease spread out by vectir insect Diaphorina citri and using infected bud in wood grafting. In infected citrus plant, two specific protein molecules with molecular weigt 16 kDa and 66 kDa are found. These protein molecules are not found in healthy citrus plant. The immunogenicity of PS16 accumulated on leaf of citrus plant infected by CVPD is known yet. The research material were leaves of citrus plant infected CVPD, leaves of healthy citrus plant and reagent used these research are for isolation of the total protein leaf of citrus plant, SDS-PAGE electroforesis, electroelution of PS16, ELISA Methods, Dot-Blot Method, anti-PS16 as aprimery antibody and secondary antibody is anti-Rabbit IgG Conjugated AP. The result of the research showed that of PS16 accumulated on leaf of citrus plant infected CVPD has immunogenic character. It is indicated by increase of the titer anti-PS16 after first immunization ang 2nd booster by indirect ELISA method and can be used to induce antibody (anti-PS16 and so showed that positive reaction between PS16 with anti-PS16. It is indicated by purples dark blue on cellulose membrane by Dot Blot method.

Detection and molecular characterization of Candidatus liberibacter spp. causing huanglongbing (HLB) in indigenous citrus cultivars in Pakistan

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Zafarullah, A.; Saleem, F.

2016-01-01

Citrus greening or huanglongbing (HLB) is one of major devastating citrus diseases all over the world. This disease is caused by fastidious ?-proteobacterium, Candidatus liberibacter spp. and is transmitted by grafting as well as psyllids Diaphorina citri or Trioza erytreae. The objective of this study was to identify and characterize the huanglongbing (HLB) infectious pathogen in commercial (Kinnow and sweet oranges) varieties by using molecular markers such as 16S rRNA, 16S/23S rRNA and outer membrane protein fragments from symptomatic leaves of assorted citrus varieties. DNA extracted from forty different citrus (including mandarin and sweet oranges) varieties having HLB-symptomatic plants from different orchards of Pakistan. Gene-specific primers for 16SrDNA, 16S/23SrDNA and outer membrane protein (OMP) gene regions were used for identification of Ca. liberibacter spp. An amplified fragment of 1174 bp from 16SrDNA, 900 bp of 16S/23S rRNA and 600 bp was observed for OMP gene fragments of Asian isolates. The resulted fragments were TA cloned and sequenced from both strands. The infectious bacterium was identified as Candidatus liberibacter asiaticus and was found in 17 samples (42%). The seasonal variation on prevalence of Candidatus liberibacter asiaticus in citrus varieties was well observed. It declined during spring season due to unfavourable temperature and humidity for Candidatus liberibacter asiaticus because disease symptoms showed mostly at low humidity and warm temperature (up to 35 degree C). (author)

Disruption of Vector Host Preference with Plant Volatiles May Reduce Spread of Insect-Transmitted Plant Pathogens.

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Martini, Xavier; Willett, Denis S; Kuhns, Emily H; Stelinski, Lukasz L

2016-05-01

Plant pathogens can manipulate the odor of their host; the odor of an infected plant is often attractive to the plant pathogen vector. It has been suggested that this odor-mediated manipulation attracts vectors and may contribute to spread of disease; however, this requires further broad demonstration among vector-pathogen systems. In addition, disruption of this indirect chemical communication between the pathogen and the vector has not been attempted. We present a model that demonstrates how a phytophathogen (Candidatus Liberibacter asiaticus) can increase its spread by indirectly manipulating the behavior of its vector (Asian citrus psyllid, Diaphorina citri Kuwayama). The model indicates that when vectors are attracted to pathogen-infected hosts, the proportion of infected vectors increases, as well as, the proportion of infected hosts. Additionally, the peak of infected host populations occurs earlier as compared with controls. These changes in disease dynamics were more important during scenarios with higher vector mortality. Subsequently, we conducted a series of experiments to disrupt the behavior of the Asian citrus psyllid. To do so, we exposed the vector to methyl salicylate, the major compound released following host infection with the pathogen. We observed that during exposure or after pre-exposure to methyl salicylate, the host preference can be altered; indeed, the Asian citrus psyllids were unable to select infected hosts over uninfected counterparts. We suggest mechanisms to explain these interactions and potential applications of disrupting herbivore host preference with plant volatiles for sustainable management of insect vectors.

Epigenetic regulation of facultative heterochromatinisation in Planococcus citri via the Me(3)K9H3-HP1-Me(3)K20H4 pathway.

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Bongiorni, Silvia; Pasqualini, Barbara; Taranta, Monia; Singh, Prim B; Prantera, Giorgio

2007-03-15

Using RNA interference (RNAi) we have conducted a functional analysis of the HP1-like chromobox gene pchet2 during embryogenesis of the mealybug Planococcus citri. Knocking down pchet2 expression results in decondensation of the male-specific chromocenter that normally arises from the developmentally-regulated facultative heterochromatinisation of the paternal chromosome complement. Together with the disappearance of the chromocenter the staining levels of two associated histone modifications, tri-methylated lysine 9 of histone H3 [Me(3)K9H3] and tri-methylated lysine 20 of histone H4 [Me(3)K20H4], are reduced to undetectable levels. Embryos treated with double-stranded RNA (dsRNA) targeting pchet2 also exhibit chromosome abnormalities, such as aberrant chromosome condensation, and also the presence of metaphases that contain 'lagging' chromosomes. We conclude that PCHET2 regulates chromosome behavior during metaphase and is a crucial component of a Me(3)K9H3-HP1-Me(3)K20H4 pathway involved in the facultative heterochromatinisation of the (imprinted) paternal chromosome set.

Characterization of Chemical Composition of Pericarpium Citri Reticulatae Volatile Oil by Comprehensive Two-Dimensional Gas Chromatography with High-Resolution Time-of-Flight Mass Spectrometry

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Kunming Qin

2013-01-01

Full Text Available Pericarpium Citri Reticulatae (Chenpi in Chinese has been widely used as an herbal medicine in Korea, China, and Japan. Chenpi extracts are used to treat indigestion and inflammatory syndromes of the respiratory tract such as bronchitis and asthma. This thesis will analyze chemical compositions of Chenpi volatile oil, which was performed by comprehensive two-dimensional gas chromatography with high-resolution time-of-flight mass spectrometry (GC × GC-HR-TOFMS. One hundred and sixty-seven components were tentatively identified, and terpene compounds are the main components of Chenpi volatile oil, a significant larger number than in previous studies. The majority of the eluted compounds, which were identified, were well separated as a result of high-resolution capability of the GC × GC method, which significantly reduces, the coelution. β-Elemene is tentatively qualified by means of GC × GC in tandem with high-resolution TOFMS detection, which plays an important role in enhancing the effects of many anticancer drugs and in reducing the side effects of chemotherapy. This study suggests that GC × GC-HR-TOFMS is suitable for routine characterization of chemical composition of volatile oil in herbal medicines.

Amostragem, caracterização de sintomas e escala diagramática da mancha graxa dos citros (Mycosphaerella citri no Recôncavo Baiano Sampling, characterization of citrus greasy spot (Mycosphaerella citri symptoms in Recôncavo Baiano and development of a diagrammatic scale for assessment of severity

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Suely Xavier de Brito Silva

2009-06-01

Full Text Available O Recôncavo da Bahia é uma região de clima propício à mancha graxa dos citros (Mycosphaerella citri, doença nunca antes estudada no Brasil. Os objetivos deste trabalho foram definir o tamanho mínimo de amostra para quantificar a incidência, caracterizar os sintomas da doença e elaborar uma escala diagramática para a avaliação de sua severidade. A partir de uma amostragem piloto realizada por avaliação de cinco folhas por quadrante, quatro quadrantes por planta e 30 plantas em cada um dos 10 pomares visitados no município de Cruz das Almas, foi determinado que o tamanho mínimo da amostra em quadrantes e folhas seria de quatro e 16 plantas, respectivamente. De 320 folhas coletadas de diferentes plantas em 11 pomares, foi constatada uma grande quantidade de lesões (média de 131 por folha sintomática, em sua maioria diminutas (média de 0,014cm². A severidade variou de 0,15% a 35,85%, com média de 7,3%. Com base na severidade real em campo, foi elaborada uma escala diagramática com seis níveis: 1%, 2%, 5%, 9%, 18% e 36% de área foliar lesionada. Cinqüenta imagens de folhas foram submetidas a avaliadores por duas vezes, os quais estimaram a severidade com e sem a utilização da escala, constatando-se um discreto benefício no grau de precisão e de acurácia das estimativas com o uso da escala.Weather conditions make the Recôncavo Region in the State of Bahia, a favorable region to the occurrence of citrus greasy spot (Mycosphaerella citri, a disease that hadn't been studied before in Brazil. Thus, this research aimed to develop a sampling method to quantify its incidence, to characterize its symptoms, and to develop a diagrammatic scale for assessing disease severity. The minimum sample size to estimate greasy spot incidence in leaves (16 plants was calculated based on a prospective sampling of five leaves per quadrant, four quadrants per tree, and 30 trees per grove, in ten groves. From 320 mature leaves collected in

Phenology of Asian citrus psyllid (Hemiptera: Liviidae) and associated parasitoids on two species of Citrus, kinnow mandarin and sweet orange, in Punjab Pakistan.

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Khan, Shouket Zaman; Arif, Muhammad Jalal; Hoddle, Christina D; Hoddle, Mark S

2014-10-01

The population phenology of Asian citrus psyllid, Diaphorina citri Kuwayama, was monitored weekly for 110 wk on two species of Citrus, kinnow mandarin and sweet orange, at two different research sites in Faisalabad, Punjab Pakistan. Citrus flush growth patterns were monitored and natural enemy surveys were conducted weekly. Flush patterns were similar for kinnow and sweet orange. However, flush on sweet orange was consistently more heavily infested with Asian citrus psyllid than kinnow flush; densities of Asian citrus psyllid eggs, nymphs, and adults were higher on sweet orange when compared with kinnow. When measured in terms of mean cumulative insect or Asian citrus psyllid days, eggs, nymphs, and adults were significantly higher on sweet orange than kinnow. Two parasitoids were recorded attacking Asian citrus psyllid nymphs, Tamarixia radiata (Waterston) and Diaphorencyrtus aligarhensis (Shafee, Alam and Agarwal). The dominant parasitoid species attacking Asian citrus psyllid nymphs on kinnow and sweet orange was T. radiata, with parasitism averaging 26%. D. aligarhensis parasitism averaged 17%. Generalist predators such as coccinellids and chrysopids were collected infrequently and were likely not important natural enemies at these study sites. Immature spiders, in particular, salticids and yellow sac spiders, were common and may be important predators of all Asian citrus psyllid life stages. Low year round Asian citrus psyllid densities on kinnow and possibly high summer temperatures, may, in part, contribute to the success of this cultivar in Punjab where Candidatus Liberibacter asiaticus, the putative causative agent of huanglongbing, a debilitating citrus disease, is widespread and vectored by Asian citrus psyllid.

Evidence for gene flow between two sympatric mealybug species (Insecta; Coccoidea; Pseudococcidae.

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Hofit Kol-Maimon

Full Text Available Occurrence of inter-species hybrids in natural populations might be evidence of gene flow between species. In the present study we found evidence of gene flow between two sympatric, genetically related scale insect species--the citrus mealybug Planococcus citri (Risso and the vine mealybug Planococcus ficus (Signoret. These species can be distinguished by morphological, behavioral, and molecular traits. We employed the sex pheromones of the two respective species to study their different patterns of male attraction. We also used nuclear ITS2 (internal transcribed spacer 2 and mitochondrial COI (Cytochrome c oxidase sub unit 1 DNA sequences to characterize populations of the two species, in order to demonstrate the outcome of a possible gene flow between feral populations of the two species. Our results showed attraction to P. ficus pheromones of all tested populations of P. citri males but not vice versa. Furthermore, ITS2 sequences revealed the presence of 'hybrid females' among P. citri populations but not among those of P. ficus. 'hybrid females' from P. citri populations identified as P. citri females according to COI sequences. We offer two hypotheses for these results. 1 The occurrence of phenotypic and genotypic traits of P. ficus in P. citri populations may be attributed to both ancient and contemporary gene flow between their populations; and 2 we cannot rule out that an ancient sympatric speciation by which P. ficus emerged from P. citri might have led to the present situation of shared traits between these species. In light of these findings we also discuss the origin of the studied species and the importance of the pherotype phenomenon as a tool with which to study genetic relationships between congener scale insects.

[Study and comparison on HPLC fingerprints of flavonoids of frequently used Chinese materia medica in citrus].

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Chen, Yonggang; Lin, Li

2011-10-01

To establish the HPLC fingerprint of flavonoids of the six clinical frequently used Chinese materia medica for regulating Qi flow,such as Citri grandis, C. grands, Citri Reticulatae Pericarpium, Citri Reticulatae Pericarpium Viride, Aurantii Fructus, and Aurantii Fructus Immaturus from Citrus, and analysis differences in the fingerprints to provide scientific basis for profile-effect research and clinical reasonable use. HPLC was performed on a C18 column with methanol-water (with acetic acid), to establish HPLC fingerprints of the six kinds of medicinal herbs on the same chromatograph condition. The six frequently used Chinese materia medica were divided into naringin type and hesperidin type according to the method of phytochemotaxonomy. Based on the retention time of chromatograph peaks, C. grandis and C. grands had fifteen common peaks; Citri Reticulatae Pericarpium, Citri Reticulatae Pericarpium Viride, Aurantii Fructus and Aurantii Fructus Immaturus had ten common peaks. All herbs had five common peaks. Compared with mutual model, the holistic similarity of chromatograms of C. grandis and C. grands was in the range of 0.9285 - 0.9962. The degree of similarity was high. For Citri Reticulatae Pericarpium, Citri Reticulatae Pericarpium Viride and Aurantii Fructus Immaturus, it was in the range of 0.9221 - 0.9973 and high. But the similarity of Aurantii Fructus was only in 0.4547 - 0.7733 with the mutual model. The established fingerprints of flavonoids of the six common traditional Chinese medicines can be used to compare the differences intuitively. Meanwhile, the peak height and peak areas of characteristic peaks are different remarkably, but whether it is connected with the different function of regulating Qi flow of the six medical materials in clinical use, is still needed to be studied.

Compatibility of Isaria fumosorosea (Hypocreales: Cordycipitaceae Blastospores with Agricultural Chemicals Used for Management of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae

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Pasco B. Avery

2013-11-01

Full Text Available Biorational insecticides are being increasingly emphasized for inclusion in integrated pest management programs for invasive insects. The entomopathogenic fungus, Isaria fumosorosea, can be used to help manage the Asian citrus psyllid with minimal impact on beneficial arthropods, but its effectiveness may be compromised by agrochemicals used to control concurrent arthropod pests and diseases. We evaluated the compatibility of I. fumosorosea blastospores with a range of spray oils and copper-based fungicides registered for use in citrus groves. Results of laboratory and greenhouse tests showed a range of responses of the fungus to the different materials, including compatibility and incompatibility. Overall, I. fumosorosea growth in vitro was reduced least by petroleum-based materials and most by botanical oils and borax, and some of the copper-based fungicides, suggesting that tank mixing of I. fumosorosea with these latter products should be avoided. However, equivalent negative effects of test materials on fungal pathogenicity were not always observed in tests with adult psyllids. We hypothesize that some oils enhanced adherence of blastospores to the insect cuticle, overcoming negative impacts on germination. Our data show that care should be taken in selecting appropriate agrochemicals for tank-mixing with commercial formulations of entomopathogenic fungi for management of citrus pests. The prospects of using I. fumosorosea for managing the invasive Asian citrus psyllid and other citrus pests are discussed.

[Rapid Identification of Epicarpium Citri Grandis via Infrared Spectroscopy and Fluorescence Spectrum Imaging Technology Combined with Neural Network].

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Pan, Sha-sha; Huang, Fu-rong; Xiao, Chi; Xian, Rui-yi; Ma, Zhi-guo

2015-10-01

To explore rapid reliable methods for detection of Epicarpium citri grandis (ECG), the experiment using Fourier Transform Attenuated Total Reflection Infrared Spectroscopy (FTIR/ATR) and Fluorescence Spectrum Imaging Technology combined with Multilayer Perceptron (MLP) Neural Network pattern recognition, for the identification of ECG, and the two methods are compared. Infrared spectra and fluorescence spectral images of 118 samples, 81 ECG and 37 other kinds of ECG, are collected. According to the differences in tspectrum, the spectra data in the 550-1 800 cm(-1) wavenumber range and 400-720 nm wavelength are regarded as the study objects of discriminant analysis. Then principal component analysis (PCA) is applied to reduce the dimension of spectroscopic data of ECG and MLP Neural Network is used in combination to classify them. During the experiment were compared the effects of different methods of data preprocessing on the model: multiplicative scatter correction (MSC), standard normal variable correction (SNV), first-order derivative(FD), second-order derivative(SD) and Savitzky-Golay (SG). The results showed that: after the infrared spectra data via the Savitzky-Golay (SG) pretreatment through the MLP Neural Network with the hidden layer function as sigmoid, we can get the best discrimination of ECG, the correct percent of training set and testing set are both 100%. Using fluorescence spectral imaging technology, corrected by the multiple scattering (MSC) results in the pretreatment is the most ideal. After data preprocessing, the three layers of the MLP Neural Network of the hidden layer function as sigmoid function can get 100% correct percent of training set and 96.7% correct percent of testing set. It was shown that the FTIR/ATR and fluorescent spectral imaging technology combined with MLP Neural Network can be used for the identification study of ECG and has the advantages of rapid, reliable effect.

UFLC-Q-TOF-MS/MS-Based Screening and Identification of Flavonoids and Derived Metabolites in Human Urine after Oral Administration of Exocarpium Citri Grandis Extract

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Xuan Zeng

2018-04-01

Full Text Available Exocarpium Citri grandis (ECG is an important Traditional Chinese Medicine (TCM for the treatment of cough and phlegm, and the flavonoids contained were considered the main effective components. To date, the systematic chemical profiling of these flavonoids and derived in vivo metabolites in human have not been well investigated. ECG was extracted using boiling water and then provided to volunteers for oral administration. Following the ingestion, urine samples were collected from volunteers over 48 h. The extract and urine samples were analyzed using ultra-fast liquid chromatography/quadrupole-time-of-flight tandem mass spectrometry (UFLC-Q-TOF-MS/MS system to screen and identify flavonoids and derived in vivo metabolites. A total of 18 flavonoids were identified in the ECG extract, and 20 metabolites, mainly glucuronide and sulfate conjugates, were screened in urine samples collected post consumption. The overall excretion of naringenin metabolites corresponded to 5.45% of intake and occurred mainly within 4–12 h after the ingestion. Meanwhile, another 29 phenolic catabolites were detected in urine. Obtained data revealed that flavonoids were abundant in the ECG extract, and these components underwent extensive phase II metabolism in humans. These results provided valuable information for further study of the pharmacology and mechanism of action of ECG.

Aspectos biológicos da fase adulta de Chrysoperla externa (Hagen, 1861 (Neuroptera: Chrysopidae oriunda de larvas alimentadas com Planococcus citri (Risso, 1813 (Hemiptera: Pseudococcidae Biological aspects of the adult stage of Chrysoperla externa (Hagen, 1861 (Neuroptera: Chrysopidae originating from the larvae fed Planococcus citri (Risso, 1813 (Hemiptera: Pseudococcidae

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Gerane Celly Dias Bezerra

2006-08-01

Full Text Available Objetivou-se estudar alguns aspectos biológicos da fase adulta de Chrysoperla externa (Hagen quando as larvas desse predador foram alimentadas com ninfas e fêmeas adultas de Planococcus citri (Risso. O experimento foi conduzido em câmara climatizada regulada a 25 ± 1 ºC, 70 ± 10% de UR e fotofase de 12 horas. Os adultos de C. externa cujas larvas foram alimentadas com ninfas nos três ínstares e fêmeas adultas da cochonilha, foram separados por sexo e 25 casais foram individualizados em gaiolas determinando-se a razão sexual, período de pré-oviposição, capacidade de oviposição diária e total durante 40 dias consecutivos, longevidade, período embrionário e viabilidade de ovos. Constatou-se que as ninfas nos três ínstares e fêmeas adultas fornecidas como alimento, não afetaram a razão sexual, período de pré-oviposição, período embrionário e longevidade dos adultos de C. externa. A produção diária e total de ovos por C. externa foi afetada pelo tipo de alimento fornecido ao predador durante a fase de larva, observando-se uma maior produção quando os casais foram alimentados na fase de larva com fêmeas adultas da cochonilha. Analisando-se a produção de ovos, observou-se uma maior concentração de oviposição nos primeiros dez dias do período reprodutivo, decrescendo nas avaliações seguintes. Foram obtidas porcentagens maiores de ovos viáveis e menores de inviáveis e inférteis, porém, não diferiram em função do alimento fornecido ao predador na fase de larva.It was aimed to study some biological aspects in the adult stage of Chrysoperla externa (Hagen when the larvae of predator were fed both nymphs and adult females of Planococcus citri (Risso. The experiment was conducted in climate chamber at 25 ± 1°C, 70 ± 10% of RH and 12-h photophase. The adults of C. externa whose larvae were fed nymphs at the three instars and adult females of mealybug were separated by sex and 25 couples were individualized

Evaluation of resistance to asiatic citrus canker among selections of pera sweet orange (Citrus sinensis)

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Asiatic citrus canker (ACC, caused by the bacterium Xanthomonas citri subsp. citri) is a destructive disease of citrus in Brazil and in several other citrus-producing countries. ACC management is problematic, and bactericides such as copper can be reasonably efficacious but do not completely control...

Resistance evaluation of Pera (Citrus sinensis) genotypes to citrus canker in greenhouse conditions

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Citrus canker, caused by the bacterium Xanthomonas citri subsp. citri results in serious yield losses and phytoregulation penalties. The use of resistant genotypes is recognized as an important tool to facilitate control of the pathogen. Studies have show that artificial inoculation results in typic...

78 FR 8435 - Importation of Fresh Citrus Fruit From Uruguay, Including Citrus Hybrids and Fortunella

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2013-02-06

...] australis, causal agent of sweet orange scab); and a pathogen (Xanthomonas citri subsp. citri, causal agent... oranges (Citrus sinensis (L.) Osbeck), lemons (C. limon (L.) Burm. f.), four species of mandarins (C... of the reading room). The PRA, titled ``Importation of Fresh Citrus Fruit, including Sweet Orange...

A plant natriuretic peptide-like molecule of the pathogen Xanthomonas axonopodis pv. citri causes rapid changes in the proteome of its citrus host

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Ottado Jorgelina

2010-03-01

Full Text Available Abstract Background Plant natriuretic peptides (PNPs belong to a novel class of peptidic signaling molecules that share some structural similarity to the N-terminal domain of expansins and affect physiological processes such as water and ion homeostasis at nano-molar concentrations. The citrus pathogen Xanthomonas axonopodis pv. citri possesses a PNP-like peptide (XacPNP uniquely present in this bacteria. Previously we observed that the expression of XacPNP is induced upon infection and that lesions produced in leaves infected with a XacPNP deletion mutant were more necrotic and lead to earlier bacterial cell death, suggesting that the plant-like bacterial PNP enables the plant pathogen to modify host responses in order to create conditions favorable to its own survival. Results Here we measured chlorophyll fluorescence parameters and water potential of citrus leaves infiltrated with recombinant purified XacPNP and demonstrate that the peptide improves the physiological conditions of the tissue. Importantly, the proteomic analysis revealed that these responses are mirrored by rapid changes in the host proteome that include the up-regulation of Rubisco activase, ATP synthase CF1 α subunit, maturase K, and α- and β-tubulin. Conclusions We demonstrate that XacPNP induces changes in host photosynthesis at the level of protein expression and in photosynthetic efficiency in particular. Our findings suggest that the biotrophic pathogen can use the plant-like hormone to modulate the host cellular environment and in particular host metabolism and that such modulations weaken host defence.

Identificação de híbridos de citros resistentes à mancha-marrom-de-alternária por meio de fAFLP e testes de patogenicidade Alternaria brown spot resistant citrus hybrid identification by means of fAFLP and pathogenicity tests

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Edvan Alves Chagas

2007-07-01

Full Text Available O objetivo deste trabalho foi identificar híbridos, oriundos de hibridações controladas entre 'Folha Murcha' x 'Ponkan' e testá-los quanto à resistência a Alternaria alternata f. sp. citri. As plântulas foram obtidas via cultura in vitro de embriões. Utilizou-se o marcador molecular fAFLP para identificação dos híbridos e, em seguida, realizou-se o teste de patogenicidade nos híbridos com isolados de Alternaria alternata f. sp. citri, em condições de laboratório. Os pares de primers EcoRI AAG - MseI CAG e EcoRI ACC - MseI CAA foram os mais eficientes na identificação dos híbridos, os quais identificaram 48,5% de híbridos. Os híbridos F64, F108, F111, F113, F131 e F139 são potencialmente resistentes a Alternaria alternata f. sp. citri.The objective of this work was to identify hybrids obtained from controlled crossings between 'Folha Murcha' x 'Ponkan', and to check their resistance to Alternaria alternata f. sp. citri. The seedlings were obtained by in vitro embryo culture. The fAFLP marker technique was used to identify the hybrids, then laboratory pathogenicity test of the hybrids was accomplished with Alternaria alternata f. sp. citri isolates. The pairs of primers EcoRI AAG - Msel CAG and EcoRI ACC - Msel CAA were the most efficient for hybrids identification, and distinguished 48.5% of hybrids. Hybrids F64, F108, F111, F113, F131 and F139 showed potential resistance to the Alternaria alternata f. sp. citri.

Chemicalome and metabolome profiling of polymethoxylated flavonoids in Citri Reticulatae Pericarpium based on an integrated strategy combining background subtraction and modified mass defect filter in a Microsoft Excel Platform.

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Zeng, Su-Ling; Duan, Li; Chen, Bai-Zhong; Li, Ping; Liu, E-Hu

2017-07-28

Detection of metabolites in complex biological matrixes is a great challenge because of the background noise and endogenous components. Herein, we proposed an integrated strategy that combined background subtraction program and modified mass defect filter (MMDF) data mining in a Microsoft Excel platform for chemicalome and metabolome profiling of the polymethoxylated flavonoids (PMFs) in Citri Reticulatae Pericarpium (CRP). The exogenously-sourced ions were firstly filtered out by the developed Visual Basic for Applications (VBA) program incorporated in the Microsoft Office. The novel MMDF strategy was proposed for detecting both target and untarget constituents and metabolites based on narrow, well-defined mass defect ranges. The approach was validated to be powerful, and potentially useful for the metabolite identification of both single compound and homologous compound mixture. We successfully identified 30 and 31 metabolites from rat biosamples after oral administration of nobiletin and tangeretin, respectively. A total of 56 PMFs compounds were chemically characterized and 125 metabolites were captured. This work demonstrated the feasibility of the integrated approach for reliable characterization of the constituents and metabolites in herbal medicines. Copyright © 2017 Elsevier B.V. All rights reserved.

Asian Citrus Psyllid Expression Profiles Suggest Candidatus Liberibacter Asiaticus-Mediated Alteration of Adult Nutrition and Metabolism, and of Nymphal Development and Immunity.

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Meenal Vyas

Full Text Available The Asian citrus psyllid (ACP Diaphorina citri Kuwayama (Hemiptera: Psyllidae is the insect vector of the fastidious bacterium Candidatus Liberibacter asiaticus (CLas, the causal agent of citrus greening disease, or Huanglongbing (HLB. The widespread invasiveness of the psyllid vector and HLB in citrus trees worldwide has underscored the need for non-traditional approaches to manage the disease. One tenable solution is through the deployment of RNA interference technology to silence protein-protein interactions essential for ACP-mediated CLas invasion and transmission. To identify psyllid interactor-bacterial effector combinations associated with psyllid-CLas interactions, cDNA libraries were constructed from CLas-infected and CLas-free ACP adults and nymphs, and analyzed for differential expression. Library assemblies comprised 24,039,255 reads and yielded 45,976 consensus contigs. They were annotated (UniProt, classified using Gene Ontology, and subjected to in silico expression analyses using the Transcriptome Computational Workbench (TCW (http://www.sohomoptera.org/ACPPoP/. Functional-biological pathway interpretations were carried out using the Kyoto Encyclopedia of Genes and Genomes databases. Differentially expressed contigs in adults and/or nymphs represented genes and/or metabolic/pathogenesis pathways involved in adhesion, biofilm formation, development-related, immunity, nutrition, stress, and virulence. Notably, contigs involved in gene silencing and transposon-related responses were documented in a psyllid for the first time. This is the first comparative transcriptomic analysis of ACP adults and nymphs infected and uninfected with CLas. The results provide key initial insights into host-parasite interactions involving CLas effectors that contribute to invasion-virulence, and to host nutritional exploitation and immune-related responses that appear to be essential for successful ACP-mediated circulative, propagative CLas

Study of entomophatogenic fungus to control vector insect of citrus tristeza virus on citrus

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Dwiastuti M.E.

2017-08-01

Full Text Available Citrus Tristeza Virus (CTV disease is a silent killer, which threatens to decrease productivity, quality and even death of citrus plants and the erosion of genetic resources. Spreading in the field very quickly by the intermediate insect vector pest, aphid (Toxoptera citricida, T. Aurantii and A. Gosypii. The microbes studied for potential biopesticide candidates are: Beauveria bassiana and Hirsutella citriformis, and Metarhizium anisopliae (Metch Sorokin previously reported to control Diaphorina citri pests resulting effectiveness of > 25% and was able to suppress yield loss up to 10%. The objectives of the study examined the effectiveness of entomopathogen in controlling the pest of CTV vector, Toxoptera citricida, in the laboratory and screen house, to findout the physiological, biochemical and molecular physiology of entomopathogen. The results showed that the best entomopathogen suspension concentration was B.bassiana 106 followed by H. citriformis 106 and M. anisopliae 106. Entomopatogen B. bassiana and H. citriformis effectively controled the CTV vector pest in the laboratory. In the semi-field experiments at the screen house, the most effective result was H.citriformis 106 and the combination of H.citriformis 106 + B.bassiana 106, killing up to 50% and 100% on day 7th H.citriformis had the most physiological character, was able to develop optimally at a temperature of 20-400C and humidity between 60-80%. The biochemical character of the entomopathogenic fungus B.bassiana contained cellulase enzyme and phosphate solvent and IAA hormone, at most compared to the others. H.citriformis had not been found to contain enzymes and hormones. The molecular biochemical characterization of entomopathogenic fungi using FS1 and NS2 primers more clearly distinguished isolates and entomopathogenic species.

Male Psyllids Differentially Learn in the Context of Copulation

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Dara G. Stockton

2017-02-01

Full Text Available In the Asian citrus psyllid, Diaphorina citri Kuwayama, stimulatory cuticular hydrocarbons act as sex pheromone attractants. Male psyllids locate aggregations of females using those olfactory cues, as well as vibrational communication on the plant surface. Although previous research has indicated that learning plays a role in modulating female reproductive behaviors in psyllids, it is unknown whether males similarly use learning to increase the likelihood of copulatory success. We used an olfactometer-based bio-assay to study the effects of experience on male response to female odor. First, we compared male attraction to female odor in virgin and previously mated males. Second, we tested the effect of several modes of experience with a novel odor, vanillin, to determine whether mating, feeding, or general environmental exposure elicited a learned response. We found that male attraction to female odor significantly increased after mating experience. In addition, we found that males learn about odor specifically in the context of mating, rather than feeding or general exposure. Electrophysiological measurements of antennal response to odorants confirmed that mating status did not affect the sensitivity of the peripheral nervous system to volatile stimuli implicating learning at the level of the central nervous system. These results suggest that male response to female odor is not an entirely innate behavior. Males may require mating experience with female conspecifics to develop attraction to those olfactory cues produced by the female and in association with the female’s habitat. This adaptive plasticity may allow males to detect females in an ever-changing environment and promote diversification and further specialization on different host genotypes.

Combining 'omics and microscopy to visualize interactions between the Asian citrus psyllid vector and the Huanglongbing pathogen Candidatus Liberibacter asiaticus in the insect gut.

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Angela Kruse

Full Text Available Huanglongbing, or citrus greening disease, is an economically devastating bacterial disease of citrus. It is associated with infection by the gram-negative bacterium Candidatus Liberibacter asiaticus (CLas. CLas is transmitted by Diaphorina citri, the Asian citrus psyllid (ACP. For insect transmission to occur, CLas must be ingested during feeding on infected phloem sap and cross the gut barrier to gain entry into the insect vector. To investigate the effects of CLas exposure at the gut-pathogen interface, we performed RNAseq and mass spectrometry-based proteomics to analyze the transcriptome and proteome, respectively, of ACP gut tissue. CLas exposure resulted in changes in pathways involving the TCA cycle, iron metabolism, insecticide resistance and the insect's immune system. We identified 83 long non-coding RNAs that are responsive to CLas, two of which appear to be specific to the ACP. Proteomics analysis also enabled us to determine that Wolbachia, a symbiont of the ACP, undergoes proteome regulation when CLas is present. Fluorescent in situ hybridization (FISH confirmed that Wolbachia and CLas inhabit the same ACP gut cells, but do not co-localize within those cells. Wolbachia cells are prevalent throughout the gut epithelial cell cytoplasm, and Wolbachia titer is more variable in the guts of CLas exposed insects. CLas is detected on the luminal membrane, in puncta within the gut epithelial cell cytoplasm, along actin filaments in the gut visceral muscles, and rarely, in association with gut cell nuclei. Our study provides a snapshot of how the psyllid gut copes with CLas exposure and provides information on pathways and proteins for targeted disruption of CLas-vector interactions at the gut interface.

The phloem-sap feeding mealybug (Ferrisia virgata carries 'Candidatus Liberibacter asiaticus' populations that do not cause disease in host plants.

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Marco Pitino

Full Text Available 'Candidatus Liberibacter asiaticus' (Las is the primary causal agent of huanglongbing (HLB, the most devastating disease of citrus worldwide. There are three known insect vectors of the HLB-associated bacteria, and all are members of the Hemiptera: Diaphorina citri (Psyllidae, Trioza erytreae (Triozidae, and Cacopsylla (Psylla citrisuga (Psyllidae. In this study, we found that another hemipteran, the striped mealybug Ferrisia virgata (Cockerell (Hemiptera: Pseudococcidae, was able to acquire and retain Las bacteria. The bacterial titers were positively correlated with the feeding acquisition time on Las-infected leaf discs, with a two-weeks feeding period resulting in Ct values ranging from 23.1 to 36.1 (8.24 × 10(7 to 1.07 × 10(4 Las cells per mealybug. We further discovered that the prophage/phage populations of Las in the mealybugs were different from those of Las in psyllids based on Las prophage-specific molecular markers: infected psyllids harbored the Las populations with prophage/phage FP1 and FP2, while infected mealybugs carried the Las populations with the iFP3 being the dominant prophage/phage. As in the psyllids, Las bacteria were shown to move through the insect gut wall to the salivary glands after being ingested by the mealybug based on a time-course quantitative polymerase chain reaction (qPCR assay of the dissected digestive systems. However, Las populations transmitted by the mealybugs did not cause disease in host plants. This is the first evidence of genetic difference among Las populations harbored by different insect vectors and difference among Las populations with respect to whether or not they cause disease in host plants.

A specific interdomain interaction preserves the structural and binding properties of the ModA protein from the phytopathogen Xanthomonas citri domain interaction and transport in ModA.

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Santacruz-Perez, Carolina; Pegos, Vanessa Rodrigues; Honorato, Rodrigo V; Verli, Hugo; Lindahl, Erik; Barbosa, João Alexandre Ribeiro Gonçalves; Balan, Andrea

2013-11-01

The periplasmic-binding proteins in ATP-binding cassette systems (ABC Transporters) are responsible for the capture and delivery of ligands to their specific transporters, triggering a series of ATP-driven conformational changes that leads to the transport of the ligand. Structurally consisting of two lobes, the proteins change conformation after interaction with the ligand. The structure of the molybdate-binding protein (ModA) from Xanthomonas citri, bound to molybdate, was previously solved by our group and an interdomain interaction, mediated by a salt bridge between K127 and D59, apparently supports the binding properties and keeps the domains closed. To determinate the importance of this interaction, we built two ModA mutants, K127S and D59A, and analysed their functional and structural properties. Based on a set of spectroscopic experiments, crystallisation trials, structure determination and molecular dynamics (MD) simulations, we showed that the salt bridge is essential to maintain the structure and binding properties. Additionally, the MD simulations revealed that this mutant adopted a more compact structure that packed down the ligand-binding pocket. From the closed bound to open structure, the positioning of the helices forming the dipole and the salt bridge are essential to induce an intermediate state. Copyright © 2013 Elsevier Inc. All rights reserved.

Analysis of transcriptome differences between resistant and susceptible strains of the citrus red mite Panonychus citri (Acari: Tetranychidae.

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Bin Liu

Full Text Available BACKGROUND: The citrus red mite is a worldwide citrus pest and a common sensitizing allergen of asthma and rhinitis. It has developed strong resistance to many registered acaricides, However, the molecular mechanisms of resistance remain unknown. we therefore used next generation sequencing technology to investigate the global transcriptomes between resistant strains and susceptible strains. RESULTS: We obtained 34,159, 30,466 and 32,217 unigenes by assembling the SS reads, RS reads and SS&RS reads respectively. There are total 17,581 annotated unigenes from SS&RS reads by BLAST searching databases of nr, the Clusters of Orthologous Groups (COGs and Kyoto Encyclopedia of Genes and Genomes (KEGG with an E-value ≤ 1e-5, in which 7,075 unigenes were annotated in the COG database, 12, 712 unigenes were found in the KEGG database and 3,812 unigenes were assigned to Gene ontology (GO. Moreover, 2,701 unigenes were judged to be the differentially expressed genes (DEGs based on the uniquely mapped reads. There are 219 pathways in all annotated unigenes and 198 pathways in DEGs that mapped to the KEGG database. We identified 211 metabolism genes and target genes related to general insecticide resistance such as P450 and Cytochrome b, and further compared their differences between RS and SS. Meanwhile, we identified 105 and 194 genes related to growth and reproduction, respectively, based on the mode of action of Hexythiazox. After further analyses, we found variation in sequences but not in gene expression related to mite growth and reproduction between different strains. CONCLUSION: To our knowledge, this is the first comparative transcriptome study to discover candidate genes involved in phytophagous mite resistance. This study identified differential unigenes related to general pesticide resistance and organism growth and reproduction in P. citri. The assembled, annotated transcriptomes provide a valuable genomic resource for further understanding

Simultaneous Quantification of Seven Bioactive Flavonoids in Citri Reticulatae Pericarpium by Ultra-Fast Liquid Chromatography Coupled with Tandem Mass Spectrometry.

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Zhao, Lian-Hua; Zhao, Hong-Zheng; Zhao, Xue; Kong, Wei-Jun; Hu, Yi-Chen; Yang, Shi-Hai; Yang, Mei-Hua

2016-05-01

Citri Reticulatae Pericarpium (CRP) is a commonly-used traditional Chinese medicine with flavonoids as the major bioactive components. Nevertheless, the contents of the flavonoids in CRP of different sources may significantly vary affecting their therapeutic effects. Thus, the setting up of a reliable and comprehensive quality assessment method for flavonoids in CRP is necessary. To set up a rapid and sensitive ultra-fast liquid chromatography coupled with tandem mass spectrometry (UFLC-MS/MS) method for simultaneous quantification of seven bioactive flavonoids in CRP. A UFLC-MS/MS method coupled to ultrasound-assisted extraction was developed for simultaneous separation and quantification of seven flavonoids including hesperidin, neohesperidin, naringin, narirutin, tangeretin, nobiletin and sinensetin in 16 batches of CRP samples from different sources in China. The established method showed good linearity for all analytes with correlation coefficient (R) over 0.9980, together with satisfactory accuracy, precision and reproducibility. Furthermore, the recoveries at the three spiked levels were higher than 89.71% with relative standard deviations (RSDs) lower than 5.19%. The results indicated that the contents of seven bioactive flavonoids in CRP varied significantly among different sources. Among the samples under study, hesperidin showed the highest contents in 16 samples ranged from 27.50 to 86.30 mg/g, the contents of hesperidin in CRP-15 and CRP-9 were 27.50 and 86.30 mg/g, respectively, while, the amount of narirutin was too low to be measured in some samples. This study revealed that the developed UFLC-MS/MS method was simple, sensitive and reliable for simultaneous quantification of multi-components in CRP with potential perspective for quality control of complex matrices. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

A plant natriuretic peptide-like molecule of the pathogen Xanthomonas axonopodis pv. citri causes rapid changes in the proteome of its citrus host

KAUST Repository

Garavaglia, Betiana S; Thomas, Ludivine; Zimaro, Tamara; Gottig, Natalia; Daurelio, Lucas D; Ndimba, Bongani; Orellano, Elena G; Ottado, Jorgelina; Gehring, Christoph A

2010-01-01

Background: Plant natriuretic peptides (PNPs) belong to a novel class of peptidic signaling molecules that share some structural similarity to the N-terminal domain of expansins and affect physiological processes such as water and ion homeostasis at nano-molar concentrations. The citrus pathogen Xanthomonas axonopodis pv. citri possesses a PNP-like peptide (XacPNP) uniquely present in this bacteria. Previously we observed that the expression of XacPNP is induced upon infection and that lesions produced in leaves infected with a XacPNP deletion mutant were more necrotic and lead to earlier bacterial cell death, suggesting that the plant-like bacterial PNP enables the plant pathogen to modify host responses in order to create conditions favorable to its own survival.Results: Here we measured chlorophyll fluorescence parameters and water potential of citrus leaves infiltrated with recombinant purified XacPNP and demonstrate that the peptide improves the physiological conditions of the tissue. Importantly, the proteomic analysis revealed that these responses are mirrored by rapid changes in the host proteome that include the up-regulation of Rubisco activase, ATP synthase CF1 ? subunit, maturase K, and ?- and ?-tubulin.Conclusions: We demonstrate that XacPNP induces changes in host photosynthesis at the level of protein expression and in photosynthetic efficiency in particular. Our findings suggest that the biotrophic pathogen can use the plant-like hormone to modulate the host cellular environment and in particular host metabolism and that such modulations weaken host defence. 2010 Garavaglia et al; licensee BioMed Central Ltd.

A plant natriuretic peptide-like molecule of the pathogen Xanthomonas axonopodis pv. citri causes rapid changes in the proteome of its citrus host

KAUST Repository

Garavaglia, Betiana S

2010-03-21

Background: Plant natriuretic peptides (PNPs) belong to a novel class of peptidic signaling molecules that share some structural similarity to the N-terminal domain of expansins and affect physiological processes such as water and ion homeostasis at nano-molar concentrations. The citrus pathogen Xanthomonas axonopodis pv. citri possesses a PNP-like peptide (XacPNP) uniquely present in this bacteria. Previously we observed that the expression of XacPNP is induced upon infection and that lesions produced in leaves infected with a XacPNP deletion mutant were more necrotic and lead to earlier bacterial cell death, suggesting that the plant-like bacterial PNP enables the plant pathogen to modify host responses in order to create conditions favorable to its own survival.Results: Here we measured chlorophyll fluorescence parameters and water potential of citrus leaves infiltrated with recombinant purified XacPNP and demonstrate that the peptide improves the physiological conditions of the tissue. Importantly, the proteomic analysis revealed that these responses are mirrored by rapid changes in the host proteome that include the up-regulation of Rubisco activase, ATP synthase CF1 ? subunit, maturase K, and ?- and ?-tubulin.Conclusions: We demonstrate that XacPNP induces changes in host photosynthesis at the level of protein expression and in photosynthetic efficiency in particular. Our findings suggest that the biotrophic pathogen can use the plant-like hormone to modulate the host cellular environment and in particular host metabolism and that such modulations weaken host defence. 2010 Garavaglia et al; licensee BioMed Central Ltd.

Citri Reticulatae Pericarpium (Chenpi): Botany, ethnopharmacology, phytochemistry, and pharmacology of a frequently used traditional Chinese medicine.

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Yu, Xin; Sun, Shuang; Guo, Yuyan; Liu, Yan; Yang, Dayu; Li, Guoyu; Lü, Shaowa

2018-06-28

Citri Reticulatae Pericarpium (Rutaceae, CRP), commonly called as Chenpi () in Chinese, is most frequently used as a qi-regulating drug in thousands of Chinese medicine prescriptions. CRP is found mainly in major citrus-producing areas such as the Guangdong, Guangxi, Sichuan, Fujian, and Zhejiang Provinces of China. Since thousands of years in China, CRP has been used widely in clinical practice to treat nausea, vomiting, indigestion, anepithymia, diarrhea, cough, expectoration, and so on. Currently, CRP is listed in the Pharmacopoeia of the People's Republic of China. The present paper reviews the botany, ethnopharmacology, phytochemistry, pharmacology, quality control, and toxicology of CRP. Information on CRP was gathered from various sources including the books on traditional Chinese herbal medicine; scientific databases including Elsevier, PubMed, and ScienceDirect; Baidu Scholar; CNKI; and others and from different professional websites. Approximately 140 chemical compounds have been isolated and identified from CRP. Among them, volatile oils and flavonoids are generally considered as the main bioactive and characteristic ingredients. CRP possesses wide pharmacological effects such as having a beneficial effect on the cardiovascular, digestive, and respiratory systems, antitumor, antioxidant, and anti-inflammatory properties; and a protective effect on the liver and nerve. Moreover, hesperidin is chosen as an indicator in the quantitative determination of CRP, and the quantity of aflatoxin in CRP must not exceed the standard limit mentioned in the pharmacopoeia. In brief, CRP has a warming nature, and hence, it can be used in harmony with a lot of medicines. CRP not only exhibits its effects individually but also aids other medicines exhibit a better effect. CRP can be consumed with tea, food, alcohol, and medicine. Irrespective of the form it is being consumed, CRP not only shows a synergistic effect but also has strengths on its own. Modern pharmacological

The effect of pre spring spray to reduce of citrus important pests.

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Hashemi, B; Damavandian, M R; Shoushtaril, R Vafaei; Tafaghodynia, B

2008-10-01

The importance of pre spring spray against citrus aphids, Pulvinaria aurantii Cockerell and Panonychus citri McGregor that are the most important pest of citrus during spring was tested. In this research, 150 trees ten years old sweet orange (Thomson navel on Citrus aurantium (root stocks)) in a citrus orchard approximately three hectares sampled. The experiment was laid out in a totally randomized (one-way) design replicated five times. According to the results, the pre spring spray do not effect on population density of citrus aphids and P. aurantii during March, April, May and June. However, the P. citri population decreased. Therefore, it seems the pre spring spray in citrus orchards is not necessary, but if P. citri is observed, the pre spring spray should be recommended.

Exogenous application of methyl jasmonate and salicylic acid on citrus foliage: Effecs on foliar volatiles and aggregation behavior of Asian citrus psyllid (Diaphorina citri)

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Methyl jasmonate (MeJA) and salicylic acid (SA) are well-known activators of chemical defenses in plants. The SA pathway is involved in citrus response to infection by Candidatus Liberibacter asiaticus (CLas); less is known about the role of jasmonates in citrus defense response. We examined the eff...

Use of micro-CT to elucidate details of the anatomy and feeding of the Asian citrus psyllid Diaphorina citri Kuwayama, 1908 (Insecta: Hemiptera, Liviidae)

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Huanglongbing (HLB), also known as citrus greening disease, is caused by plant-infecting bacteria. The most prominent pathogen within the Americas: United States of America, Mexico, and Brazil, is Candidatus Liberibacter asiaticus, which affects plants of the Family: Rutaceae, in particularly citrus...

Toxicity of bifenazate and its principal active metabolite, diazene, to Tetranychus urticae and Panonychus citri and their relative toxicity to the predaceous mites, Phytoseiulus persimilis and Neoseiulus californicus.

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Ochiai, Noriaki; Mizuno, Masayuki; Mimori, Norihiko; Miyake, Toshihiko; Dekeyser, Mark; Canlas, Liza Jara; Takeda, Makio

2007-01-01

Bifenazate is a novel carbazate acaricide discovered by Uniroyal Chemical (now Chemtura Corporation) for the control of phytophagous mites infesting agricultural and ornamental crops. Its acaricidal activity and that of its principal active metabolite, diazene, were characterized. Bifenazate and diazene had high toxicity and specificity both orally and topically to all life stages of Tetranychus urticae and Panonychus citri. Acute poisoning was observed with no temperature dependency. No cross-resistance was found to mites resistant to several other classes of acaricides, such as tebufenpyrad, etoxazole, fenbutatin oxide and dicofol. Bifenazate remained effective for a long time with only about a 10% loss of efficacy on T. urticae after 1 month of application in the field. All stages of development of the predatory mites, Phytoseiulus persimilis and Neoseiulus californicus, survived treatment by both bifenazate and diazene. When adult females of the two predatory mite species were treated with either bifenazate or diazene, they showed a normal level of fecundity and predatory activity in the laboratory, effectively suppressing spider mite population growth. Even when the predators were fed spider mite eggs that had been treated previously with bifenazate, they survived. These findings indicate that bifenazate is a very useful acaricide giving high efficacy, long-lasting activity and excellent selectivity for spider mites. It is, therefore, concluded that bifenazate is an ideal compound for controlling these pest mites.

Sampling Methods for Detection and Monitoring of the Asian Citrus Psyllid (Hemiptera: Psyllidae).

Science.gov (United States)

Monzo, C; Arevalo, H A; Jones, M M; Vanaclocha, P; Croxton, S D; Qureshi, J A; Stansly, P A

2015-06-01

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama is a key pest of citrus due to its role as vector of citrus greening disease or "huanglongbing." ACP monitoring is considered an indispensable tool for management of vector and disease. In the present study, datasets collected between 2009 and 2013 from 245 citrus blocks were used to evaluate precision, sensitivity for detection, and efficiency of five sampling methods. The number of samples needed to reach a 0.25 standard error-mean ratio was estimated using Taylor's power law and used to compare precision among sampling methods. Comparison of detection sensitivity and time expenditure (cost) between stem-tap and other sampling methodologies conducted consecutively at the same location were also assessed. Stem-tap sampling was the most efficient sampling method when ACP densities were moderate to high and served as the basis for comparison with all other methods. Protocols that grouped trees near randomly selected locations across the block were more efficient than sampling trees at random across the block. Sweep net sampling was similar to stem-taps in number of captures per sampled unit, but less precise at any ACP density. Yellow sticky traps were 14 times more sensitive than stem-taps but much more time consuming and thus less efficient except at very low population densities. Visual sampling was efficient for detecting and monitoring ACP at low densities. Suction sampling was time consuming and taxing but the most sensitive of all methods for detection of sparse populations. This information can be used to optimize ACP monitoring efforts. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Mechanism of action and selective toxicity of ascamycin, a nucleoside antibiotic.

OpenAIRE

Osada, H; Isono, K

1985-01-01

An unidentified Streptomyces sp. produces two nucleoside antibiotics, ascamycin and its dealanyl derivative. In contrast to the broad antibacterial activity of dealanylascamycin against various gram-negative and gram-positive bacteria, ascamycin showed selective toxicity against Xanthomonas citri and X. oryzae. Both ascamycin and dealanylascamycin inhibited the protein synthesis of X. citri, but only dealanylascamycin inhibited that of Escherichia coli. In cell-free systems from E. coli and X...

Effect of whole yeast cell product supplementation (CitriStim®) on immune responses and cecal microflora species in pullet and layer chickens during an experimental coccidial challenge.

Science.gov (United States)

Markazi, Ashley D; Perez, Victor; Sifri, Mamduh; Shanmugasundaram, Revathi; Selvaraj, Ramesh K

2017-07-01

Three separate experiments were conducted to study the effects of whole yeast cell product supplementation in pullets and layer hens. Body weight gain, fecal and intestinal coccidial oocyst counts, cecal microflora species, cytokine mRNA amounts, and CD4+ and CD8+ T-cell populations in the cecal tonsils were analyzed following an experimental coccidial infection. In Experiment I, day-old Leghorn layer chicks were fed 3 experimental diets with 0, 0.1, or 0.2% whole yeast cell product (CitriStim®, ADM, Decatur, IL). At 21 d of age, birds were challenged with 1 × 105 live coccidial oocysts. Supplementation with whole yeast cell product decreased the fecal coccidial oocyst count at 7 (P = 0.05) and 8 (P product and challenged with 1 × 105 live coccidial oocysts on d 25 of whole yeast cell product feeding. Supplementation with whole yeast cell product decreased the coccidial oocyst count in the intestinal content (P product increased relative proportion of Lactobacillus (P product decreased CD8+ T cell percentages (P product and challenged with 1 × 105 live coccidial oocysts on d 66 of whole yeast cell product feeding. At 5 d post-coccidial challenge, whole yeast cell product supplementation down-regulated (P = 0.01) IL-10 mRNA amount. It could be concluded that supplementing whole yeast cell product can help minimize coccidial infection in both growing pullets and layer chickens. © 2017 Poultry Science Association Inc.

Rapid establishment of a regular distribution of adult tropical Drosophila parasitoids in a multi-patch environment by patch defence behaviour.

Science.gov (United States)

de Jong, Peter W; Hemerik, Lia; Gort, Gerrit; van Alphen, Jacques J M

2011-01-01

Females of the larval parasitoid of Drosophila, Asobara citri, from sub-Saharan Africa, defend patches with hosts by fighting and chasing conspecific females upon encounter. Females of the closely related, palearctic species Asobara tabida do not defend patches and often search simultaneously in the same patch. The effect of patch defence by A. citri females on their distribution in a multi-patch environment was investigated, and their distributions were compared with those of A. tabida. For both species 20 females were released from two release-points in replicate experiments. Females of A. citri quickly reached a regular distribution across 16 patches, with a small variance/mean ratio per patch. Conversely, A. tabida females initially showed a clumped distribution, and after gradual dispersion, a more Poisson-like distribution across patches resulted (variance/mean ratio was closer to 1 and higher than for A. citri). The dispersion of A. tabida was most probably an effect of exploitation: these parasitoids increasingly made shorter visits to already exploited patches. We briefly discuss hypotheses on the adaptive significance of patch defence behaviour or its absence in the light of differences in the natural history of both parasitoid species, notably the spatial distribution of their hosts.

Whitefly Pest Species (Homoptera: Aleyrodidae) on Citrus Trees

OpenAIRE

Katja Žanić; Sonja Kačić; Miro Katalinić

2000-01-01

Today, the Citrus whitefly, Dialeurodes citri (Ashmead), is a very important pest on all Citrus species throughout the citrus growing areas in Croatia. It causes direct damage by sucking the plant juice from the leaves. Furthermore, immatures excrete honeydew that stimulates sooy mold. The presence of sooty mold on contaminated leaves interferes with the photosynthesis of plants. Citrus fruits coated by sooty mold lose its market value. Because Dialeurodes citri is poorly known in Croatia, th...

Three Phoma spp. synthesised novel silver nanoparticles that possess excellent antimicrobial efficacy.

Science.gov (United States)

Rai, Mahendra; Ingle, Avinash P; Gade, Aniket K; Duarte, Marta Cristina Teixeira; Duran, Nelson

2015-10-01

The authors report extracellular mycosynthesis of silver nanoparticles (AgNPs) by Phoma capsulatum, Phoma putaminum and Phoma citri. The AgNPs thus synthesised were characterised by UV-visible spectrophotometer, Fourier transform infrared spectroscopy, Nanosight LM20 and transmission electron microscopy, which confirmed the synthesis of mostly spherical and polydisperse nanoparticles capped with proteins. The size of AgNPs was found in the range of 10-80 , 5-80 and 5-90 nm with an average size of 31.85, 25.43 and 23.29 nm by P. capsulatum, P. putaminum and P. citri, respectively. Further, potential antimicrobial activity was reported against Aspergillus niger, Candida albicans, Salmonella choleraesuis, Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli. The lowest minimal inhibitory concentration (MIC) (0.85 µg/ml) was reported for AgNPs synthesised from P. citri against S. choleraesuis. However, AgNPs synthesised from P. capsulatum showed the highest MIC (10.62 µg/ml) against S. choleraesuis, P. aeruginosa and E. coli (clinical isolate). The same MIC values (10.62 µg/ml) were also reported against P. aeruginosa and both clinical and standard isolates of E. coli for AgNPs synthesised from P. citri. It was also observed that all the silver nanoparticles showed remarkable antifungal and antibacterial activity against these tested pathogens as compared with the commercially available antifungal and antibacterial agents.

The effect of nutritional spray programs applied to mitigate symptoms of Huanglongbing on fruit drop caused by HLB and citrus canker on ‘Hamlin’ orange trees

OpenAIRE

Roberts, P. D.; Rouse, R. E.; Teems, S. S.; Sytsma, R. E.; Shobert, Z.

2014-01-01

Huanglongbing (HLB) was detected in Florida in 2005 and has reached 100% incidence in certain citrus plantings in southwest Florida. The putative causal agent of HLB in Florida is the bacterium Candidatus Liberibacter asiaticus (CLa).  Citrus canker caused by the bacterium Xanthomonas citri subsp. citri is endemic in Florida.  In 2011 and 2012, fruit drop on young ‘Hamlin’ trees with symptoms of HLB and/or citrus canker was particularly severe, with more than 90% fruit drop recorded. Nutritio...

An Ecoinformatics Approach to Field-Scale Evaluation of Insecticide Effects in California Citrus: Are Citrus Thrips and Citrus Red Mite Induced Pests?

Science.gov (United States)

Livingston, George; Hack, Lindsey; Steinmann, Kimberly P; Grafton-Cardwell, Elizabeth E; Rosenheim, Jay A

2018-05-28

Experimental approaches to studying the consequences of pesticide use, including impacts on beneficial insects, are vital; however, they can be limited in scale and realism. We show that an ecoinformatics approach that leverages existing data on pesticides, pests, and beneficials across multiple fields can provide complementary insights. We do this using a multi-year dataset (2002-2013) on pesticide applications and density estimates of two pests, citrus thrips (Scirtothrips citri (Moulton [Thysanoptera: Thripidae])) and citrus red mites (Panonychus citri McGregor [Acari: Tetranychidae]), and a natural enemy (Euseius spp. predatory mites) collected from citrus groves in the San Joaquin Valley of California. Using correlative analyses, we investigated the long-term consequences of pesticide use on S. citri and P. citri population densities to evaluate the hypothesis that the pest status of these species is largely due to the disruption of natural biological control-i.e., these are induced pests. We also evaluated short-term pesticide efficacy (suppression of citrus thrips and citrus red mite populations immediately post-application) and asked if it was correlated with the suppression of Euseius predator populations. Although the short-term efficacy of different pesticides varied significantly, our dataset does not suggest that the use of citrus pesticides suppressed Euseius densities or worsened pest problems. We also find that there is no general trade-off between pesticide efficacy and pesticide risk to Eusieus, such that highly effective and minimally disruptive compounds were available to citrus growers during the studied time period.

Isolation of radiation resistant fungal strains from highly radioactive field

International Nuclear Information System (INIS)

Adam, Y.M.; Aziz, N.H.; Attaby, H.S.H.

1995-01-01

This study examined the radiation resistance of fungal flora isolated from the hot-lab around the radiation sources, cobalt 137 and radium 226 . The predominant mould species were: Aspergillus flavus, A. Niger, penicillium chrysogenum, cladosporium herbarum, fusarium oxysporum and alternaria citri. The D 10 values of F. Oxysporum; 2.00 KGy, A. Flavus; 1.40 KGy, P. chrysogenum; 1.15 KGy, and A. citri; 0.95 KGy, are about 1.67, 3.10, 1.92 and 1.36 folds as the D 1 0 values of the same isolates recovered from soil

Economic injury levels for Asian citrus psyllid control in process oranges from mature trees with high incidence of huanglongbing.

Directory of Open Access Journals (Sweden)

Cesar Monzo

Full Text Available The Asian citrus psyllid (ACP, Diaphorina citri Kuwayama, is the key pest of citrus wherever it occurs due to its role as vector of huanglongbing (HLB also known as citrus greening disease. Insecticidal vector control is considered to be the primary strategy for HLB management and is typically intense owing to the severity of this disease. While this approach slows spread and also decreases severity of HLB once the disease is established, economic viability of increasingly frequent sprays is uncertain. Lacking until now were studies evaluating the optimum frequency of insecticide applications to mature trees during the growing season under conditions of high HLB incidence. We related different degrees of insecticide control with ACP abundance and ultimately, with HLB-associated yield losses in two four-year replicated experiments conducted in commercial groves of mature orange trees under high HLB incidence. Decisions on insecticide applications directed at ACP were made by project managers and confined to designated plots according to experimental design. All operational costs as well as production benefits were taken into account for economic analysis. The relationship between management costs, ACP abundance and HLB-associated economic losses based on current prices for process oranges was used to determine the optimum frequency and timing for insecticide applications during the growing season. Trees under the most intensive insecticidal control harbored fewest ACP resulting in greatest yields. The relationship between vector densities and yield loss was significant but differed between the two test orchards, possibly due to varying initial HLB infection levels, ACP populations or cultivar response. Based on these relationships, treatment thresholds during the growing season were obtained as a function of application costs, juice market prices and ACP densities. A conservative threshold for mature trees with high incidence of HLB would help

A eukaryotic-acquired gene by a biotrophic phytopathogen allows prolonged survival on the host by counteracting the shut-down of plant photosynthesis.

Science.gov (United States)

Garavaglia, Betiana S; Thomas, Ludivine; Gottig, Natalia; Dunger, Germán; Garofalo, Cecilia G; Daurelio, Lucas D; Ndimba, Bongani; Orellano, Elena G; Gehring, Chris; Ottado, Jorgelina

2010-01-28

Xanthomonas citri pv. citri, the bacteria responsible for citrus canker posses a biological active plant natriuretic peptide (PNP)-like protein, not present in any other bacteria. PNPs are a class of extracellular, systemically mobile peptides that elicit a number of plant responses important in homeostasis and growth. Previously, we showed that a Xanthomonas citri pv. citri mutant lacking the PNP-like protein XacPNP produced more necrotic lesions in citrus leaves than wild type infections and suggested a role for XacPNP in the regulation of host homeostasis. Here we have analyzed the proteome modifications observed in citrus leaves infected with the wild type and XacPNP deletion mutant bacteria. While both of them cause down-regulation of enzymes related to photosynthesis as well as chloroplastic ribosomal proteins, proteins related to defense responses are up-regulated. However, leaves infiltrated with the XacPNP deletion mutant show a more pronounced decrease in photosynthetic proteins while no reduction in defense related proteins as compared to the wild-type pathogen. This suggests that XacPNP serves the pathogen to maintain host photosynthetic efficiency during pathogenesis. The results from the proteomics analyses are consistent with our chlorophyll fluorescence data and transcript analyses of defense genes that show a more marked reduction in photosynthesis in the mutant but no difference in the induction of genes diagnostic for biotic-stress responses. We therefore conclude that XacPNP counteracts the shut-down of host photosynthesis during infection and in that way maintains the tissue in better conditions, suggesting that the pathogen has adapted a host gene to modify its natural host and render it a better reservoir for prolonged bacterial survival and thus for further colonization.

ORF Alignment: NC_003919 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Xanthomonas axonopodis pv. citri str. 306] ... Length = 97 ... Query: 99 ... VARAMQYIDTHLYEPVSVAHLAQAACMSRFHFARVFRDAVGASPM...EYLRLRRLERAQAML 158 ... VARAMQYIDTHLYEPVSVAHLAQAACMSRFHFARVFRDAVGASPME...YLRLRRLERAQAML Sbjct: 1 ... VARAMQYIDTHLYEPVSVAHLAQAACMSRFHFARVFRDAVGASPMEYLRLRRLERAQAML 60 ...

ORF Alignment: NC_003919 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available as ... axonopodis pv. citri str. 306] ... Length = 221 ... Query: 40 ... VLATDAHGYLENQQLPDSVQLVPPPPQ...DGSAALANDEAVAKAMLALRGTPRWELAAQDAV 99 ... VLATDAHGYLENQQLPDSVQLVPPPPQDGSAAL...ANDEAVAKAMLALRGTPRWELAAQDAV Sbjct: 3 ... VLATDAHGYLENQQLPDSVQLVPPPPQDGSAALANDEAVAKAMLALRGTPRWELAAQDAV 62 ... Quer

ORF Alignment: NC_003919 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available AC4077 ... [Xanthomonas axonopodis pv. citri str. 306] ... Length = 150 ... Query: 744 VRPVRPEDDA...LFRAFFARVTDEDLRLRFFQSVKHFSHEFIARLTQLDYARSIALVAIDPR 803 ... VRPVRPEDDALFRAF...FARVTDEDLRLRFFQSVKHFSHEFIARLTQLDYARSIALVAIDPR Sbjct: 1 ... VRPVRPEDDALFRAFFARVTDEDLRLRFFQSVKHFSHEFIARLTQLDYARS

ORF Alignment: NC_003919 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... pv. citri str. 306] ... Length = 255 ... Query: 6 ... KKTALVTGASSGMGKAIAHRLIQDGYQVYVAARNVDRMADLALLGAHPLC...MDVSRDDDIV 65 ... KKTALVTGASSGMGKAIAHRLIQDGYQVYVAARNVDRMADLALLGAHPLC...MDVSRDDDIV Sbjct: 1 ... KKTALVTGASSGMGKAIAHRLIQDGYQVYVAARNVDRMADLALLGAHPLCMDVSRDDDIV 60 ... Query: 126 KRAGHIVN

ORF Alignment: NC_003919 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available tein ... B [Xanthomonas axonopodis pv. citri str. 306] ... Length = 101 ... Query: 9 ... TSHRPYLLRALVEWIND...NGMTPHVLVDAGLPGVQVPASAVKDGRVVLNIAERAVVGLQVD 68 ... TSHRPYLLRALVEWIND...NGMTPHVLVDAGLPGVQVPASAVKDGRVVLNIAERAVVGLQVD Sbjct: 1 ... TSHRPYLLRALVEWINDNGMTPHVLVDAGLPGVQVPASAVKDGRVVLNIAERAVVGLQVD 60 ...

ORF Alignment: NC_006834 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available tein ... B [Xanthomonas axonopodis pv. citri str. 306] ... Length = 101 ... Query: 9 ... TSHRPYLLRALVEWIND...NGMTPHVLVDAGLPGVQVPASAVKDGRVVLNIAERAVVGLQVD 68 ... TSHRPYLLRALVEWIND...NGMTPHVLVDAGLPGVQVPASAVKDGRVVLNIAERAVVGLQVD Sbjct: 1 ... TSHRPYLLRALVEWINDNGMTPHVLVDAGLPGVQVPASAVKDGRVVLNIAERAVVGLQVD 60 ...

A eukaryotic-acquired gene by a biotrophic phytopathogen allows prolonged survival on the host by counteracting the shut-down of plant photosynthesis

KAUST Repository

Garavaglia, Betiana S.

2010-01-28

Xanthomonas citri pv. citri, the bacteria responsible for citrus canker posses a biological active plant natriuretic peptide (PNP)-like protein, not present in any other bacteria. PNPs are a class of extracellular, systemically mobile peptides that elicit a number of plant responses important in homeostasis and growth. Previously, we showed that a Xanthomonas citri pv. citri mutant lacking the PNP-like protein XacPNP produced more necrotic lesions in citrus leaves than wild type infections and suggested a role for XacPNP in the regulation of host homeostasis. Here we have analyzed the proteome modifications observed in citrus leaves infected with the wild type and XacPNP deletion mutant bacteria. While both of them cause downregulation of enzymes related to photosynthesis as well as chloroplastic ribosomal proteins, proteins related to defense responses are up-regulated. However, leaves infiltrated with the XacPNP deletion mutant show a more pronounced decrease in photosynthetic proteins while no reduction in defense related proteins as compared to the wild-type pathogen. This suggests that XacPNP serves the pathogen to maintain host photosynthetic efficiency during pathogenesis. The results from the proteomics analyses are consistent with our chlorophyll fluorescence data and transcript analyses of defense genes that show a more marked reduction in photosynthesis in the mutant but no difference in the induction of genes diagnostic for biotic-stress responses. We therefore conclude that XacPNP counteracts the shut-down of host photosynthesis during infection and in that way maintains the tissue in better conditions, suggesting that the pathogen has adapted a host gene to modify its natural host and render it a better reservoir for prolonged bacterial survival and thus for further colonization. 2010 Garavaglia et al.

A eukaryotic-acquired gene by a biotrophic phytopathogen allows prolonged survival on the host by counteracting the shut-down of plant photosynthesis.

Directory of Open Access Journals (Sweden)

Betiana S Garavaglia

Full Text Available Xanthomonas citri pv. citri, the bacteria responsible for citrus canker posses a biological active plant natriuretic peptide (PNP-like protein, not present in any other bacteria. PNPs are a class of extracellular, systemically mobile peptides that elicit a number of plant responses important in homeostasis and growth. Previously, we showed that a Xanthomonas citri pv. citri mutant lacking the PNP-like protein XacPNP produced more necrotic lesions in citrus leaves than wild type infections and suggested a role for XacPNP in the regulation of host homeostasis. Here we have analyzed the proteome modifications observed in citrus leaves infected with the wild type and XacPNP deletion mutant bacteria. While both of them cause down-regulation of enzymes related to photosynthesis as well as chloroplastic ribosomal proteins, proteins related to defense responses are up-regulated. However, leaves infiltrated with the XacPNP deletion mutant show a more pronounced decrease in photosynthetic proteins while no reduction in defense related proteins as compared to the wild-type pathogen. This suggests that XacPNP serves the pathogen to maintain host photosynthetic efficiency during pathogenesis. The results from the proteomics analyses are consistent with our chlorophyll fluorescence data and transcript analyses of defense genes that show a more marked reduction in photosynthesis in the mutant but no difference in the induction of genes diagnostic for biotic-stress responses. We therefore conclude that XacPNP counteracts the shut-down of host photosynthesis during infection and in that way maintains the tissue in better conditions, suggesting that the pathogen has adapted a host gene to modify its natural host and render it a better reservoir for prolonged bacterial survival and thus for further colonization.

A eukaryotic-acquired gene by a biotrophic phytopathogen allows prolonged survival on the host by counteracting the shut-down of plant photosynthesis

KAUST Repository

Garavaglia, Betiana S.; Thomas, Ludivine; Gottig, Natalia; Dunger, Germá n; Garofalo, Cecilia G.; Daurelio, Lucas D.; Ndimba, Bongani; Orellano, Elena G.; Gehring, Christoph A; Ottado, Jorgelina

2010-01-01

Xanthomonas citri pv. citri, the bacteria responsible for citrus canker posses a biological active plant natriuretic peptide (PNP)-like protein, not present in any other bacteria. PNPs are a class of extracellular, systemically mobile peptides that elicit a number of plant responses important in homeostasis and growth. Previously, we showed that a Xanthomonas citri pv. citri mutant lacking the PNP-like protein XacPNP produced more necrotic lesions in citrus leaves than wild type infections and suggested a role for XacPNP in the regulation of host homeostasis. Here we have analyzed the proteome modifications observed in citrus leaves infected with the wild type and XacPNP deletion mutant bacteria. While both of them cause downregulation of enzymes related to photosynthesis as well as chloroplastic ribosomal proteins, proteins related to defense responses are up-regulated. However, leaves infiltrated with the XacPNP deletion mutant show a more pronounced decrease in photosynthetic proteins while no reduction in defense related proteins as compared to the wild-type pathogen. This suggests that XacPNP serves the pathogen to maintain host photosynthetic efficiency during pathogenesis. The results from the proteomics analyses are consistent with our chlorophyll fluorescence data and transcript analyses of defense genes that show a more marked reduction in photosynthesis in the mutant but no difference in the induction of genes diagnostic for biotic-stress responses. We therefore conclude that XacPNP counteracts the shut-down of host photosynthesis during infection and in that way maintains the tissue in better conditions, suggesting that the pathogen has adapted a host gene to modify its natural host and render it a better reservoir for prolonged bacterial survival and thus for further colonization. 2010 Garavaglia et al.

77 FR 59709 - Citrus Greening and Asian Citrus Psyllid; Quarantine and Interstate Movement Regulations

Science.gov (United States)

2012-10-01

... occur. The pathogen can also be transmitted by two insect vectors in the family Psyllidae: Diaphorina... California due to the presence of Asian citrus psyllid (ACP), a vector of the bacterial pathogen that causes... INFORMATION: Background Citrus greening, also known as Huanglongbing disease of citrus, is considered to be...

Exogenous application of the plant signalers methyl jasmonate & salicylic acid induces changes in volatile emissions from citrus foliage & influences the aggregation behavior of ACP (Diaphorina citri), vector of Huanglongbing

Science.gov (United States)

Huanglongbing or citrus greening is a destructive disease that threatens citrus production worldwide; it is putatively caused by the phloem-limited bacterium Candidatus Liberibacter asiaticus (Las). Currently the disease is untreatable and control efforts focus on intensive insecticide use to contro...

ORF Alignment: NC_003919 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... axonopodis pv. citri str. 306] ... Length = 131 ... Query: 1 ... MNVLIREATEGDAAAIELLTMVAFMRAERSRHDEHQVIAGLRQDGALALSLVADHD...GYVV 60 ... MNVLIREATEGDAAAIELLTMVAFMRAERSRHDEHQVIAGLRQDGALALSLVADHD...GYVV Sbjct: 1 ... MNVLIREATEGDAAAIELLTMVAFMRAERSRHDEHQVIAGLRQDGALALSLVADHDGYVV 60 ... Query: 121 AFFRRLGFAVE 131 ... AFFRRLGFAVE Sbjct: 121 AFFRRLGFAVE 131

ORF Alignment: NC_003919 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ter ... [Xanthomonas axonopodis pv. citri str. 306] ... Length = 125 ... Query: 66 ... ELTVGDVMISRSQMVSLPVEARFID...LMKQVVESGHSRFPVHGENKDEVLGILLAKDLLRG 125 ... ELTVGDVMISRSQMVSLPVEARFID...LMKQVVESGHSRFPVHGENKDEVLGILLAKDLLRG Sbjct: 1 ... ELTVGDVMISRSQMVSLPVEARFIDLMKQVVESGHSRFPVHGENKDEVLGILLAKDLLRG 60 ... Query: 186 EQIVG 190 ... EQIVG Sbjct: 121 EQIVG 125

ORF Alignment: NC_006834 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ter ... [Xanthomonas axonopodis pv. citri str. 306] ... Length = 125 ... Query: 72 ... ELTVGDVMISRSQMVSLPVEARFID...LMKQVVESGHSRFPVHGENKDEVLGILLAKDLLRG 131 ... ELTVGDVMISRSQMVSLPVEARFID...LMKQVVESGHSRFPVHGENKDEVLGILLAKDLLRG Sbjct: 1 ... ELTVGDVMISRSQMVSLPVEARFIDLMKQVVESGHSRFPVHGENKDEVLGILLAKDLLRG 60 ... Query: 192 EQIVG 196 ... EQIVG Sbjct: 121 EQIVG 125

Vulnerability and behavioral response to ultraviolet radiation in the components of a foliar mite prey-predator system

Science.gov (United States)

Tachi, Fuyuki; Osakabe, Masahiro

2012-12-01

Ambient ultraviolet-B (UVB) radiation impacts plant-dwelling arthropods including herbivorous and predatory mites. However, the effects of UVB on prey-predator systems, such as that between the herbivorous spider mite and predatory phytoseiid mite, are poorly understood. A comparative study was conducted to determine the vulnerability and behavioral responses of these mites to ultraviolet (UV) radiation. First, we analyzed dose-response (cumulative irradiance-mortality) curves for the eggs of phytoseiid mites ( Neoseiulus californicus, Neoseiulus womersleyi, and Phytoseiulus persimilis) and the spider mite ( Tetranychus urticae) to UVB radiation from a UV lamp. This indicated that the phytoseiid mites were more vulnerable than the spider mite, although P. persimilis was slightly more tolerant than the other two phytoseiid mites. Second, we compared the avoidance behavior of adult female N. californicus and two spider mite species ( T. urticae, a lower leaf surface user; Panonychus citri, an upper leaf surface user) in response to solar UV and visible light. N. californicus actively avoided both types of radiation, whereas P. citri showed only minimal avoidance behavior. T. urticae actively avoided UV as well as N. californicus but exhibited a slow response to visible light as well as P. citri. Such variation in vulnerability and avoidance behavior accounts for differences in the species adaptations to solar UVB radiation. This may be the primary factor determining habitat use among these mites on host plant leaves, subsequently affecting accessibility by predators and also intraguild competition.

ORF Alignment: NC_003919 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_003919 gi|21240843 >1p6rA 5 81 4 80 8e-10 ... gb|AAM34961.1| methicillin resistanc...e protein [Xanthomonas axonopodis pv. citri ... str. 306] ref|NP_640425.1| methicillin resistance ...

ORF Alignment: NC_003921 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_003921 gi|21264213 >1ub4C 1 63 1 63 4e-10 ... gb|AAM39232.1| plasmid stable inheritance... protein I [Xanthomonas axonopodis pv. ... citri str. 306] ref|NP_644714.1| plasmid stable ... inheritance

ORF Alignment: NC_003921 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_003921 gi|21264212 >1m1fA 1 109 2 110 1e-34 ... gb|AAM39231.1| plasmid stable inheritance... protein K [Xanthomonas axonopodis pv. ... citri str. 306] ref|NP_644713.1| plasmid stable ... inheritance

The type III protein secretion system contributes to Xanthomonas citri subsp. citri biofilm formation

KAUST Repository

Zimaro, Tamara; Thomas, Ludivine; Marondedze, Claudius; Sgro, Germá n G; Garofalo, Cecilia G; Ficarra, Florencia A; Gehring, Christoph A; Ottado, Jorgelina; Gottig, Natalia

2014-01-01

Background: Several bacterial plant pathogens colonize their hosts through the secretion of effector proteins by a Type III protein secretion system (T3SS). The role of T3SS in bacterial pathogenesis is well established but whether this system

Overexpression of a modified plant thionin enhances disease resistance to citrus canker and huanglongbing (HLB, citrus greening)

Science.gov (United States)

Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the United States citrus industry. Citrus canker is also an economically important disease associated with a bacterial pathogen (Xanthomonas citri). In this study, we characterized e...

NCBI nr-aa BLAST: CBRC-TNIG-22-0185 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-TNIG-22-0185 ref|NP_642362.1| putative manganese transport protein MntH [Xanth...omonas axonopodis pv. citri str. 306] sp|Q8PKX1|MNTH_XANAC Probable manganese transport protein mntH gb|AAM36898.1| manga

A TALE of transposition: Tn3-like transposons play a major role in the spread of pathogenicity determinants of Xanthomonas citri and other xanthomonads.

Science.gov (United States)

Ferreira, Rafael Marini; de Oliveira, Amanda Carolina P; Moreira, Leandro M; Belasque, José; Gourbeyre, Edith; Siguier, Patricia; Ferro, Maria Inês T; Ferro, Jesus A; Chandler, Michael; Varani, Alessandro M

2015-02-17

Members of the genus Xanthomonas are among the most important phytopathogens. A key feature of Xanthomonas pathogenesis is the translocation of type III secretion system (T3SS) effector proteins (T3SEs) into the plant target cells via a T3SS. Several T3SEs and a murein lytic transglycosylase gene (mlt, required for citrus canker symptoms) are found associated with three transposition-related genes in Xanthomonas citri plasmid pXAC64. These are flanked by short inverted repeats (IRs). The region was identified as a transposon, TnXax1, with typical Tn3 family features, including a transposase and two recombination genes. Two 14-bp palindromic sequences within a 193-bp potential resolution site occur between the recombination genes. Additional derivatives carrying different T3SEs and other passenger genes occur in different Xanthomonas species. The T3SEs include transcription activator-like effectors (TALEs). Certain TALEs are flanked by the same IRs as found in TnXax1 to form mobile insertion cassettes (MICs), suggesting that they may be transmitted horizontally. A significant number of MICs carrying other passenger genes (including a number of TALE genes) were also identified, flanked by the same TnXax1 IRs and delimited by 5-bp target site duplications. We conclude that a large fraction of T3SEs, including individual TALEs and potential pathogenicity determinants, have spread by transposition and that TnXax1, which exhibits all of the essential characteristics of a functional transposon, may be involved in driving MIC transposition. We also propose that TALE genes may diversify by fork slippage during the replicative Tn3 family transposition. These mechanisms may play a crucial role in the emergence of Xanthomonas pathogenicity. Xanthomonas genomes carry many insertion sequences (IS) and transposons, which play an important role in their evolution and architecture. This study reveals a key relationship between transposons and pathogenicity determinants in

Survey of current crop management practices in a mixed-ricefield landscape, Mekong Delta, Vietnam - potential of habitat manipulation for improved control of citrus leafminer and citrus red mite

NARCIS (Netherlands)

Mele, van P.; Lenteren, van J.C.

2002-01-01

In the Mekong Delta, Vietnam, the citrus leafminer Phyllocnistis citrella (CLM) and the citrus red mite Panonychus citri are major pests in both sweet orange (Citrus sinensis) and Tieu mandarin (C. reticulata). Survey data indicate that these pest problems might be aggravated after farmers have

Resistance to pathogens in terpene down-regulated orange fruits inversely correlates with the accumulation of D-limonene in peel oil glands.

Science.gov (United States)

Rodríguez, Ana; Shimada, Takehiko; Cervera, Magdalena; Redondo, Ana; Alquézar, Berta; Rodrigo, María Jesús; Zacarías, Lorenzo; Palou, Lluís; López, María M; Peña, Leandro

2015-01-01

Volatile organic compounds (VOCs) are secondary metabolites acting as a language for the communication of plants with the environment. In orange fruits, the monoterpene D-limonene accumulates at very high levels in oil glands from the peel. Drastic down-regulation of D-limonene synthase gene expression in the peel of transgenic oranges harboring a D-limonene synthase transgene in antisense (AS) configuration altered the monoterpene profile in oil glands, mainly resulting in reduced accumulation of D-limonene. This led to fruit resistance against Penicillium digitatum (Pd), Xanthomonas citri subsp. citri (Xcc) and other specialized pathogens. Here, we analyze resistance to pathogens in independent AS and empty vector (EV) lines, which have low, medium or high D-limonene concentrations and show that the level of resistance is inversely related to the accumulation of D-limonene in orange peels, thus explaining the need of high D-limonene accumulation in mature oranges in nature for the efficient attraction of specialized microorganism frugivores.

Browse Title Index

African Journals Online (AJOL)

Items 4701 - 4750 of 11090 ... ... Evaluating the effect of some botanical insecticides on the citrus mealybug Planococcus citri (Risso) (Hemiptera: Pseudococcidae), Abstract PDF ... Vol 6, No 22 (2007), Evaluation of in vitro antiviral activity of a brown alga (Cystoseira myrica) from the Persian Gulf against herpes simplex virus ...

Identifying the impacts of climate change on key pests and diseases of plant and animal industries

International Nuclear Information System (INIS)

Luck, Jo; Aurambout, Jean-Philippe; Finlay, Kyla; Azuloas, Joe; Constable, Fiona; Rijswijk, Bonny Rowles-Van

2007-01-01

temperature data, coupled to plant physiology data and pest growth and population data for the Asian citrus psyllid (Diaphorina citri), vector of citrus greening disease, to determine the potential distribution and abundance of the insect in relation to climate change. This model will be built on to provide a more accurate prediction of the effects of climate change on plant and animal biosecurity and to develop contingency plans for government and industry to respond to and minimise the risks

Modeling the incidence of citrus canker in leaves of the sweet orange variety ‘Pera’

Directory of Open Access Journals (Sweden)

Danielle da Silva Pompeu

2017-01-01

Full Text Available Citrus canker, caused by the bacterium Xanthomonas citri subsp. citri, is one of the most important diseases of citrus. The use of resistant genotypes plays an important role in the management and control of the disease and is the most environmentally sustainable approach to disease control. Citrus canker incidence was recorded in an experiment on nine genotypes of the sweet orange variety ‘Pera’ grafted on four rootstocks. The experiment was started in 2010 and the incidence of citrus canker on the leaves was recorded on a quarterly basis. The incidence data from the experiment were analyzed using a zero-inflated Beta regression model (RBIZ, which is the appropriate method to describe data with large numbers of zeros. Based on the residual analysis, the data fit the model well. The discrete component of the explanatory variable, rootstock, was not significant as a factor affecting the onset of disease, in contrast with the continuous component, genotype, which was significant in explaining the incidence of citrus canker.

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that the bacterium was Xanthomonas campestris pv citri A and the disease was a form of citrus canker. The disease incidence ... declined from 88.3% to 1.7% in six years (2009-2014) without the application of any control measure. It was conjectured that .... on yeast nutrient agar in plates using a heat- sterilized inoculation ...

Preclinical Mouse Models of Neurofibromatosis

Science.gov (United States)

2005-11-01

Genet. 20, 207–211.347 A R T I C L EPlaat, B.E., Molenaar , W.M., Mastik, M.F., Hoekstra, H.J., te Meerman, G.J., and van den Berg, E. (1999). Computer...predict tumor recurrence. Am. J. Surg. Pathol. 28: 101–106. Shelly, M., Mosesson, Y., Citri, A., Lavi, S., Zwang, Y., Mela- med- Book , N., Aroeti, B., and

Ectopic Expression of Xylella fastidiosa rpfF Conferring Production of Diffusible Signal Factor in Transgenic Tobacco and Citrus Alters Pathogen Behavior and Reduces Disease Severity.

Science.gov (United States)

Caserta, R; Souza-Neto, R R; Takita, M A; Lindow, S E; De Souza, A A

2017-11-01

The pathogenicity of Xylella fastidiosa is associated with its ability to colonize the xylem of host plants. Expression of genes contributing to xylem colonization are suppressed, while those necessary for insect vector acquisition are increased with increasing concentrations of diffusible signal factor (DSF), whose production is dependent on RpfF. We previously demonstrated that transgenic citrus plants ectopically expressing rpfF from a citrus strain of X. fastidiosa subsp. pauca exhibited less susceptibility to Xanthomonas citri subsp. citri, another pathogen whose virulence is modulated by DSF accumulation. Here, we demonstrate that ectopic expression of rpfF in both transgenic tobacco and sweet orange also confers a reduction in disease severity incited by X. fastidiosa and reduces its colonization of those plants. Decreased disease severity in the transgenic plants was generally associated with increased expression of genes conferring adhesiveness to the pathogen and decreased expression of genes necessary for active motility, accounting for the reduced population sizes achieved in the plants, apparently by limiting pathogen dispersal through the plant. Plant-derived DSF signal molecules in a host plant can, therefore, be exploited to interfere with more than one pathogen whose virulence is controlled by DSF signaling.

Cyber Security: A Crisis of Prioritization

Science.gov (United States)

2005-02-01

Society (CITRIS) and Professor University of California, Berkeley J. Carter Beese , Jr. President Riggs Capital Partners Pedro Celis, Ph.D. Software...Science University of North Carolina at Chapel Hill William J. Hannigan President AT&T Jonathan C. Javitt, M.D., M.P.H. Senior Fellow Potomac Institute...CHAIR F. Thomson Leighton MEMBERS J. Carter Beese , Jr. Patricia Thomas Evans Luis E. Fiallo Harold Mortazavian David A. Patterson Alice G

Coffea arabica L., a new host plant for Acetobacter diazotrophicus, and isolation of other nitrogen-fixing acetobacteria.

OpenAIRE

Jimenez-Salgado, T; Fuentes-Ramirez, L E; Tapia-Hernandez, A; Mascarua-Esparza, M A; Martinez-Romero, E; Caballero-Mellado, J

1997-01-01

Acetobacter diazotrophicus was isolated from coffee plant tissues and from rhizosphere soils. Isolation frequencies ranged from 15 to 40% and were dependent on soil pH. Attempts to isolate this bacterial species from coffee fruit, from inside vesicular-arbuscular mycorrhizal fungi spores, or from mealybugs (Planococcus citri) associated with coffee plants were not successful. Other acid-producing diazotrophic bacteria were recovered with frequencies of 20% from the coffee rhizosphere. These N...

Computational Science: Ensuring America’s Competitiveness

Science.gov (United States)

2005-06-01

Interest of Society (CITRIS) and Professor University of California, Berkeley J. Carter Beese , Jr. President Riggs Capital Partners Pedro Celis, Ph.D...and Library Science University of North Carolina at Chapel Hill William J. Hannigan President AT&T Jonathan C. Javitt, M.D., M.P.H. Senior Fellow...Vannier, Ph.D., Professor of Radiology, University of Chicago • Jonathan C. Silverstein, M.D., M.S., FACS, Assistant Professor of Surgery, University of

Artificial Neural Network Analysis of Xinhui Pericarpium Citri ...

African Journals Online (AJOL)

Methods: Artificial neural networks (ANN) models, including general regression neural network (GRNN) and multi-layer ... N-hexane (HPLC grade) was purchased from. Fisher Scientific. ..... Simultaneous Quantification of Seven Flavonoids in.

Inhibition of Xanthomonas citri subsp. citri, causal agent of citrus canker, by a strain of Zymomonas mobilis mobilis isolated from sugarcane

OpenAIRE

Romero, María E.; Ramallo, C. Jacqueline; Ploper, L. Daniel

2008-01-01

Zymomonas mobilis mobilis (Zm) produce factores antimicrobianos que actúan sobre un amplio espectro de microorganismos patógenos para el hombre, animales y plantas. Un problema importante a resolver en los tratamientos con antimicrobianos, es el desarrollo de resistencia a compuestos empleados actualmente, no siendo las bacterias fitopatógenas una excepción. En el presente trabajo se realizaron ensayos de antagonismo con células (pruebas de estrías cruzadas) y sobrenadantes concentrados (Sc) ...

Influence of gamma radiation on antibodies fixation on polystyrene. Application to ELISA enzyme immunoassay

International Nuclear Information System (INIS)

Esterlin, S.

1986-01-01

This thesis is divided into two parts: the first part includes a description of the ELISA test, a comparative analysis of the main supports (polystyrene microtitration trays) used for this technique and an evaluation of gamma radiation effects on the quality of the supports. The study was carried out with the following antigens: Spiroplasma citri R8A2, a tymovirus and Tristeza virus. The second part dealt with the effects of gamma radiations on antibodies and antibody-support system (interaction immobilized immunoglobulins-plastic support) [fr

Molecular and Morphological Identification of Mealybug Species (Hemiptera: Pseudococcidae) in Brazilian Vineyards

Science.gov (United States)

Pacheco da Silva, Vitor C.; Bertin, Aline; Blin, Aurélie; Germain, Jean-François; Bernardi, Daniel; Rignol, Guylène; Botton, Marcos; Malausa, Thibaut

2014-01-01

Mealybugs (Hemiptera: Pseudococcidae) are pests constraining the international trade of Brazilian table grapes. They damage grapes by transmitting viruses and toxins, causing defoliation, chlorosis, and vigor losses and favoring the development of sooty mold. Difficulties in mealybug identification remain an obstacle to the adequate management of these pests. In this study, our primary aim was to identify the principal mealybug species infesting the major table grape-producing regions in Brazil, by morphological and molecular characterization. Our secondary aim was to develop a rapid identification kit based on species-specific Polymerase Chain Reactions, to facilitate the routine identification of the most common pest species. We surveyed 40 sites infested with mealybugs and identified 17 species: Dysmicoccus brevipes (Cockerell), Dysmicoccus sylvarum Williams and Granara de Willink, Dysmicoccus texensis (Tinsley), Ferrisia cristinae Kaydan and Gullan, Ferrisia meridionalis Williams, Ferrisia terani Williams and Granara de Willink, Phenacoccus baccharidis Williams, Phenacoccus parvus Morrison, Phenacoccus solenopsis Tinsley, Planococcus citri (Risso), Pseudococcus viburni (Signoret), Pseudococcus cryptus Hempel, four taxa closely related each of to Pseudococcus viburni, Pseudococcus sociabilis Hambleton, Pseudococcus maritimus (Ehrhorn) and Pseudococcus meridionalis Prado, and one specimen from the genus Pseudococcus Westwood. The PCR method developed effectively identified five mealybug species of economic interest on grape in Brazil: D. brevipes, Pl. citri, Ps. viburni, Ph. solenopsis and Planococcus ficus (Signoret). Nevertheless, it is not possible to assure that this procedure is reliable for taxa that have not been sampled already and might be very closely related to the target species. PMID:25062012

Paternal inheritance in mealybugs (Hemiptera: Coccoidea: Pseudococcidae)

Science.gov (United States)

Kol-Maimon, Hofit; Mendel, Zvi; Franco, José Carlos; Ghanim, Murad

2014-10-01

Mealybugs have a haplodiploid reproduction system, with paternal genome elimination (PGE); the males are diploid soon after fertilization, but during embryogenesis, the male paternal set of chromosomes becomes heterochromatic (HC) and therefore inactive. Previous studies have suggested that paternal genes can be passed on from mealybug males to their sons, but not necessarily by any son, to the next generation. We employed crosses between two mealybug species— Planococcus ficus (Signoret) and Planococcus citri (Risso)—and between two populations of P. ficus, which differ in their mode of pheromone attraction, in order to demonstrate paternal inheritance from males to F2 through F1 male hybrids. Two traits were monitored through three generations: mode of male pheromone attraction (pherotype) and sequences of the internal transcribed spacer 2 (ITS2) gene segment (genotype). Our results demonstrate that paternal inheritance in mealybugs can occur from males to their F2 offspring, through F1 males (paternal line). F2 backcrossed hybrid males expressed paternal pherotypes and ITS2 genotypes although their mother originated through a maternal population. Further results revealed other, hitherto unknown, aspects of inheritance in mealybugs, such as that hybridization between the two species caused absence of paternal traits in F2 hybrid females produced by F1 hybrid females. Furthermore, hybridization between the two species raised the question of whether unattracted males have any role in the interactions between P. ficus and P. citri.

Antibiotics from bacillus subtilis AECL69 8. isolation and purification of a complex of antibacterial antibiotics x

International Nuclear Information System (INIS)

Ahmad, M.S.; Malik, M.A.; Shaukat, G.A.

1996-01-01

A bacterial strain bacillus subtilis AECL69 produces two anti bacterial antibiotics in a specified complex or synthetic medium. One of the antibiotics is characteristically active against Xanthomonas citri. Procedures have been described to isolate and purify a complex of xanthmonas antibiotics from the fermented complex broths, and from the fermented synthetic medium as well. Paper chromatography coupled with bioautography has shown that the complex of xanthomonas antibiotics has at least three components. The three components were indicated irrespective of the fact whether it was isolated from the fermented complex or synthetic broth. (author)

Controle químico do cancro cítrico em plantas jovens sob manejo convencional e orgânico Quemical control of citrus canker in young plants managed in conventional and organic systems

Directory of Open Access Journals (Sweden)

Otto Carlos Koller

2006-08-01

Full Text Available O objetivo deste trabalho foi avaliar o desenvolvimento de porta-enxertos de Poncirus trifoliata Raf. e de enxertos de laranjeira "Valência" (Citrus sinensis Osb., os índices de ataque de cancro cítrico, causado pela bactéria Xanthomonas citri pv. citri, artificialmente inoculada, e o controle dessa moléstia, com pulverizações cúpricas em sistemas de cultivo, convencional e orgânico, no Centro de Formação da EMATER, situado no município de Montenegro, Estado do Rio Grande do Sul. Foram testadas pulverizações cúpricas a cada 7, 14 e 21 dias, com concentrações de 0,15 e 0,30% de cobre metálico, utilizando calda bordalesa no sistema orgânico e oxicloreto de cobre no sistema convencional. Foram avaliados o crescimento do diâmetro do caule das plantas, a produção de matéria seca da parte aérea dos porta-enxertos, o número de folhas com lesões de cancro cítrico e o número de lesões presentes por folha atacada. Verificou-se que em ambos os sistemas de cultivo, no convencional e no orgânico, o desenvolvimento dos porta-enxertos foi semelhante, mas os enxertos cresceram mais no sistema convencional. Com elevada presença de fontes de inóculo, os tratamentos cúpricos não controlaram o cancro cítrico nos porta-enxertos. Já nos enxertos, com baixa presença de fontes de inóculo, tanto a calda bordalesa como o oxicloreto de cobre controlaram a doença, com melhor resposta na concentração de 0,3% de cobre metálico, aplicada em intervalos de 14 dias. Para melhorar o controle do cancro cítrico, com calda bordalesa, em sistemas orgânicos de cultivo, devem ser desenvolvidas medidas eficazes de controle da larva-minadora.The objective of the present work was to evaluate the development of the citrus rootstock Poncirus trifoliata, severity of citrus canker, caused by Xanthomonas citri pv. citri and the control of this disease with copper sprays, in conventional and organic management systems, at the Centro de Formação da

Insights into xanthomonas axonopodis pv. Citri biofilm through proteomics

KAUST Repository

Zimaro, Tamara; Thomas, Ludivine; Marondedze, Claudius; Garavaglia, Betiana S; Gehring, Christoph A; Ottado, Jorgelina; Gottig, Natalia

2013-01-01

in adherence were over-expressed, while a polynucleotide phosphorylase that was demonstrated to negatively control biofilm formation in E. coli was down-regulated. In addition, several proteins involved in protein synthesis, folding and stabilization were up

TAL effectors target the C-terminal domain of RNA polymerase II (CTD by inhibiting the prolyl-isomerase activity of a CTD-associated cyclophilin.

Directory of Open Access Journals (Sweden)

Mariane Noronha Domingues

Full Text Available Transcriptional activator-like (TAL effectors of plant pathogenic bacteria function as transcription factors in plant cells. However, how TAL effectors control transcription in the host is presently unknown. Previously, we showed that TAL effectors of the citrus canker pathogen Xanthomonas citri, named PthAs, targeted the citrus protein complex comprising the thioredoxin CsTdx, ubiquitin-conjugating enzymes CsUev/Ubc13 and cyclophilin CsCyp. Here we show that CsCyp complements the function of Cpr1 and Ess1, two yeast cyclophilins that regulate transcription by the isomerization of proline residues of the regulatory C-terminal domain (CTD of RNA polymerase II. We also demonstrate that CsCyp, CsTdx, CsUev and four PthA variants interact with the citrus CTD and that CsCyp co-immunoprecipitate with the CTD in citrus cell extracts and with PthA2 transiently expressed in sweet orange epicotyls. The interactions of CsCyp with the CTD and PthA2 were inhibited by cyclosporin A (CsA, a cyclophilin inhibitor. Moreover, we present evidence that PthA2 inhibits the peptidyl-prolyl cis-trans isomerase (PPIase activity of CsCyp in a similar fashion as CsA, and that silencing of CsCyp, as well as treatments with CsA, enhance canker lesions in X. citri-infected leaves. Given that CsCyp appears to function as a negative regulator of cell growth and that Ess1 negatively regulates transcription elongation in yeast, we propose that PthAs activate host transcription by inhibiting the PPIase activity of CsCyp on the CTD.

Molecular and morphological identification of mealybug species (Hemiptera: Pseudococcidae in Brazilian vineyards.

Directory of Open Access Journals (Sweden)

Vitor C Pacheco da Silva

Full Text Available Mealybugs (Hemiptera: Pseudococcidae are pests constraining the international trade of Brazilian table grapes. They damage grapes by transmitting viruses and toxins, causing defoliation, chlorosis, and vigor losses and favoring the development of sooty mold. Difficulties in mealybug identification remain an obstacle to the adequate management of these pests. In this study, our primary aim was to identify the principal mealybug species infesting the major table grape-producing regions in Brazil, by morphological and molecular characterization. Our secondary aim was to develop a rapid identification kit based on species-specific Polymerase Chain Reactions, to facilitate the routine identification of the most common pest species. We surveyed 40 sites infested with mealybugs and identified 17 species: Dysmicoccus brevipes (Cockerell, Dysmicoccus sylvarum Williams and Granara de Willink, Dysmicoccus texensis (Tinsley, Ferrisia cristinae Kaydan and Gullan, Ferrisia meridionalis Williams, Ferrisia terani Williams and Granara de Willink, Phenacoccus baccharidis Williams, Phenacoccus parvus Morrison, Phenacoccus solenopsis Tinsley, Planococcus citri (Risso, Pseudococcus viburni (Signoret, Pseudococcus cryptus Hempel, four taxa closely related each of to Pseudococcus viburni, Pseudococcus sociabilis Hambleton, Pseudococcus maritimus (Ehrhorn and Pseudococcus meridionalis Prado, and one specimen from the genus Pseudococcus Westwood. The PCR method developed effectively identified five mealybug species of economic interest on grape in Brazil: D. brevipes, Pl. citri, Ps. viburni, Ph. solenopsis and Planococcus ficus (Signoret. Nevertheless, it is not possible to assure that this procedure is reliable for taxa that have not been sampled already and might be very closely related to the target species.

RESPOSTA FUNCIONAL DA JOANINHA CRYPTOLAEMUS PREDANDO COCHONILHA BRANCA EM DIFERENTES TEMPERATURAS E SUBSTRATOS VEGETAIS

Directory of Open Access Journals (Sweden)

SERGIO ANTONIO DE BORTOLI

2014-01-01

Full Text Available This work was carried out to evaluate the functional response of Cryptolaemus montrouzieri Mulsant, 1850 (Coleoptera: Coccinellidae fed with Planococcus citri Risso, 1813 (Hemiptera: Pseudococcidae reared on a pumpkin hybrid (Cucurbita maxima x Cucurbita moscata (Cucurbitaceae, seedlings of Rangpur lime (Citrus limonia Rutaceae and potato (Solanum tuberosum (Solanaceae at two temperatures. The predation rate of C. montrouzieri was measured using Petri dishes of 15 cm diameter with 1, 2, 4, 8, 16 and 24 adults of P. Citri. One third instar larva, one fourfh instar and one newly emerged adult (without differentiation of sex of C. montrouzieri were added to each plate. The study was conducted in climatic chambers at temperatures of 25 and 30 º C and photophase of 12 hours. The predation rate was evaluated after 24 hours of prey exposition to the predator, by counting the number of preys trapped in the different treatments and control. The statistical design was completely randomized with four treatments x 6 subplots with 7 repetitions, the two temperatures. The values obtained were subjected to analysis of variance, to relate the number of scales preyed by larvae and adults of C. montrouzieri set up in different substrates. The amount of prey consumed by larvae and adults of the predator increased with increasing the prey density until it reaches a plateau, characterizing functional response type II. In general, the number of scales preyed by larvae and adults of C. montrouzieri was higher on potato and under temperature of 30 °C.

Studies on chemical protectors against radiation, 26

International Nuclear Information System (INIS)

Sato, Yushi; Ohta, Setsuko; Sakurai, Nobuko; Shinoda, Masato

1989-01-01

The protective potency against skin injury on mice induced by X-irradiation was studied by use of 72 extracts of crude drugs. The protective potency was determined according to the degrees of skin injury after irradiation of 1100 R, 30 kVp soft X-ray. As a result of this study, 16 kinds of crude drugs such as Rosae Fructus, Aloe arboresces (Herba), Citri Leiocarpae Exocarpium, Schizonepetae Spica, Evodiae Fructus, Bupleuri Radix, Corni Fructus, Perillae Herba, Anemarrhenae Rhizoma, Menthae Herba, Trapae Fructus, Angelicae Dahuricae Radix, Sinomeni Caulis et Rhizoma, Ephedrae Herba. Acer nikoense (Cortex), Forsythiae Fructus, revealed protective potencies on skin injury. (author)

The dual nature of trehalose in citrus canker disease: a virulence factor for Xanthomonas citri subsp. citri and a trigger for plant defence responses

KAUST Repository

Piazza, A.; Zimaro, T.; Garavaglia, B. S.; Ficarra, F. A.; Thomas, L.; Marondedze, C.; Feil, R.; Lunn, J. E.; Gehring, Christoph A; Ottado, J.; Gottig, N.

2015-01-01

and proteomic analyses of infected leaves showed that infection with XccΔotsA triggered only weak defence responses in the plant compared with infection with Xcc, and had less impact on the host plant's metabolism than the wild-type strain. These results

Ecological genomics in Xanthomonas: the nature of genetic adaptation with homologous recombination and host shifts

KAUST Repository

Huang, Chao-Li; Pu, Pei-Hua; Huang, Hao-Jen; Sung, Huang-Mo; Liaw, Hung-Jiun; Chen, Yi-Min; Chen, Chien-Ming; Huang, Ming-Ban; Osada, Naoki; Gojobori, Takashi; Pai, Tun-Wen; Chen, Yu-Tin; Hwang, Chi-Chuan; Chiang, Tzen-Yuh

2015-01-01

Background: Comparative genomics provides insights into the diversification of bacterial species. Bacterial speciation usually takes place with lasting homologous recombination, which not only acts as a cohering force between diverging lineages but brings advantageous alleles favored by natural selection, and results in ecologically distinct species, e.g., frequent host shift in Xanthomonas pathogenic to various plants. Results: Using whole-genome sequences, we examined the genetic divergence in Xanthomonas campestris that infected Brassicaceae, and X. citri, pathogenic to a wider host range. Genetic differentiation between two incipient races of X. citri pv. mangiferaeindicae was attributable to a DNA fragment introduced by phages. In contrast to most portions of the genome that had nearly equivalent levels of genetic divergence between subspecies as a result of the accumulation of point mutations, 10% of the core genome involving with homologous recombination contributed to the diversification in Xanthomonas, as revealed by the correlation between homologous recombination and genomic divergence. Interestingly, 179 genes were under positive selection; 98 (54.7%) of these genes were involved in homologous recombination, indicating that foreign genetic fragments may have caused the adaptive diversification, especially in lineages with nutritional transitions. Homologous recombination may have provided genetic materials for the natural selection, and host shifts likely triggered ecological adaptation in Xanthomonas. To a certain extent, we observed positive selection nevertheless contributed to ecological divergence beyond host shifting. Conclusion: Altogether, mediated with lasting gene flow, species formation in Xanthomonas was likely governed by natural selection that played a key role in helping the deviating populations to explore novel niches (hosts) or respond to environmental cues, subsequently triggering species diversification. © Huang et al.

Ecological genomics in Xanthomonas: the nature of genetic adaptation with homologous recombination and host shifts

KAUST Repository

Huang, Chao-Li

2015-03-15

Background: Comparative genomics provides insights into the diversification of bacterial species. Bacterial speciation usually takes place with lasting homologous recombination, which not only acts as a cohering force between diverging lineages but brings advantageous alleles favored by natural selection, and results in ecologically distinct species, e.g., frequent host shift in Xanthomonas pathogenic to various plants. Results: Using whole-genome sequences, we examined the genetic divergence in Xanthomonas campestris that infected Brassicaceae, and X. citri, pathogenic to a wider host range. Genetic differentiation between two incipient races of X. citri pv. mangiferaeindicae was attributable to a DNA fragment introduced by phages. In contrast to most portions of the genome that had nearly equivalent levels of genetic divergence between subspecies as a result of the accumulation of point mutations, 10% of the core genome involving with homologous recombination contributed to the diversification in Xanthomonas, as revealed by the correlation between homologous recombination and genomic divergence. Interestingly, 179 genes were under positive selection; 98 (54.7%) of these genes were involved in homologous recombination, indicating that foreign genetic fragments may have caused the adaptive diversification, especially in lineages with nutritional transitions. Homologous recombination may have provided genetic materials for the natural selection, and host shifts likely triggered ecological adaptation in Xanthomonas. To a certain extent, we observed positive selection nevertheless contributed to ecological divergence beyond host shifting. Conclusion: Altogether, mediated with lasting gene flow, species formation in Xanthomonas was likely governed by natural selection that played a key role in helping the deviating populations to explore novel niches (hosts) or respond to environmental cues, subsequently triggering species diversification. © Huang et al.

In vitro control of Alternaria citri using antifungal potentials of ...

African Journals Online (AJOL)

msc06-08

2012-05-24

May 24, 2012 ... black-pod disease (Phytophthora palmivora) of cacao. Biol. Control,. 50:143-149. Hanhong B (2011). Trichoderma species as abiotic and biotic stress · quenchers in plants. Res. J. Biotechnol., 6(3): 73-79. Harman GE (2000). Myths and dogmas of biocontrol: Changes in perceptions derived from research ...

Unaspis lansivora sp. n. (Hemiptera: Diaspididae), a new pest of Lansium domesticum (Meliaceae), and a key to Unaspis species.

Science.gov (United States)

Watson, Gillian W

2015-01-13

Since 2004, an undescribed species of Unaspis (Hemiptera: Diaspididae) has become a damaging pest on Lansium domesticum Corrêa in the Philippines. Its attack on the leaves causes premature senescence and defoliation, resulting in the production of few, underdeveloped, sour fruit and sometimes killing the trees. The scale was misidentified initially as Lepidosaphes ulmi (Linnaeus) and then as Unaspis citri (Comstock), but further study indicated that it was an undescribed species of potential plant quarantine significance. The pest is described as U. lansivora sp. n. and an identification key to all 19 species of Unaspis is provided. Its distribution, host range and prospects for its biological control are discussed.

Decay and acceptability of mangos treated with combinations of hot water, imazalil, and gamma radiation

International Nuclear Information System (INIS)

Spalding, D.H.; Reeder, W.F.

1986-01-01

Combination treatments with radiation at 200 or 750 Gy and hot water (53 C) or hot 0.1% a.i. imazalil (53 C) for 3 min were more effective than single treatments for control of anthracnose and stem-end rot of Tommy Atkins mangos caused by Colletotrichum gloeosporioides and Diplodia natalensis or Phomopsis citri, respectively. Irradiation at 750 Gy inhibited development of ripe skin color and caused some browning and pitting of the skin. Effects of radiation on skin color and injury were partially offset when heat treatment preceded irradiation. Individual wrapping of mangos in shrink film resulted in increased decay and breakdown. (author)

Decay and acceptability of mangos treated with combinations of hot water, imazalil, and gamma radiation

Energy Technology Data Exchange (ETDEWEB)

Spalding, D. H.; Reeder, W. F.

1986-12-15

Combination treatments with radiation at 200 or 750 Gy and hot water (53 C) or hot 0.1% a.i. imazalil (53 C) for 3 min were more effective than single treatments for control of anthracnose and stem-end rot of Tommy Atkins mangos caused by Colletotrichum gloeosporioides and Diplodia natalensis or Phomopsis citri, respectively. Irradiation at 750 Gy inhibited development of ripe skin color and caused some browning and pitting of the skin. Effects of radiation on skin color and injury were partially offset when heat treatment preceded irradiation. Individual wrapping of mangos in shrink film resulted in increased decay and breakdown. (author)

Genomic survey of pathogenicity determinants and VNTR markers in the cassava bacterial pathogen Xanthomonas axonopodis pv. Manihotis strain CIO151.

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Arrieta-Ortiz, Mario L; Rodríguez-R, Luis M; Pérez-Quintero, Álvaro L; Poulin, Lucie; Díaz, Ana C; Arias Rojas, Nathalia; Trujillo, Cesar; Restrepo Benavides, Mariana; Bart, Rebecca; Boch, Jens; Boureau, Tristan; Darrasse, Armelle; David, Perrine; Dugé de Bernonville, Thomas; Fontanilla, Paula; Gagnevin, Lionel; Guérin, Fabien; Jacques, Marie-Agnès; Lauber, Emmanuelle; Lefeuvre, Pierre; Medina, Cesar; Medina, Edgar; Montenegro, Nathaly; Muñoz Bodnar, Alejandra; Noël, Laurent D; Ortiz Quiñones, Juan F; Osorio, Daniela; Pardo, Carolina; Patil, Prabhu B; Poussier, Stéphane; Pruvost, Olivier; Robène-Soustrade, Isabelle; Ryan, Robert P; Tabima, Javier; Urrego Morales, Oscar G; Vernière, Christian; Carrere, Sébastien; Verdier, Valérie; Szurek, Boris; Restrepo, Silvia; López, Camilo; Koebnik, Ralf; Bernal, Adriana

2013-01-01

Xanthomonas axonopodis pv. manihotis (Xam) is the causal agent of bacterial blight of cassava, which is among the main components of human diet in Africa and South America. Current information about the molecular pathogenicity factors involved in the infection process of this organism is limited. Previous studies in other bacteria in this genus suggest that advanced draft genome sequences are valuable resources for molecular studies on their interaction with plants and could provide valuable tools for diagnostics and detection. Here we have generated the first manually annotated high-quality draft genome sequence of Xam strain CIO151. Its genomic structure is similar to that of other xanthomonads, especially Xanthomonas euvesicatoria and Xanthomonas citri pv. citri species. Several putative pathogenicity factors were identified, including type III effectors, cell wall-degrading enzymes and clusters encoding protein secretion systems. Specific characteristics in this genome include changes in the xanthomonadin cluster that could explain the lack of typical yellow color in all strains of this pathovar and the presence of 50 regions in the genome with atypical nucleotide composition. The genome sequence was used to predict and evaluate 22 variable number of tandem repeat (VNTR) loci that were subsequently demonstrated as polymorphic in representative Xam strains. Our results demonstrate that Xanthomonas axonopodis pv. manihotis strain CIO151 possesses ten clusters of pathogenicity factors conserved within the genus Xanthomonas. We report 126 genes that are potentially unique to Xam, as well as potential horizontal transfer events in the history of the genome. The relation of these regions with virulence and pathogenicity could explain several aspects of the biology of this pathogen, including its ability to colonize both vascular and non-vascular tissues of cassava plants. A set of 16 robust, polymorphic VNTR loci will be useful to develop a multi-locus VNTR analysis

DEVELOPMENT OF TECHNOLOGICAL SOLUTIONS FOR THE BEVERAGE INDUSTRY PREVENTIVE ORIENTATION

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N. V. Nepovinnykh

2014-01-01

Full Text Available Summary. Market demand for foods with low fat content has been expanding, but for low-fat foods good organoleptic properties need new food ingredients and solutions. Food hydrocolloids, polysaccharides and milk proteins, now are widely used in various industries, including the dairy industry, performing various functions: to thicken aqueous solutions, foaming and stabilizing foams and others. We studied the functional and technological properties and developed technology of new types of oxygencontaining beverages (smoothies on the basis of cheese whey, natural fruit and berry juices and purees and dietary fibres «Citri-Fi», including non-starch polysaccharides (guar gum and xanthan gum, contributing to the formation of a specific texture and drink as stabilizers oxygen foam. When creating new kinds of smoothies performed construction flavored beverage profile, the definition of rational parameters of preparation and entering dietary fibres; selection of the optimal concentration of dietary fiber for the formation of the desired consistency (texture drink; study of quality and safety of new types of drinks and justification expiration dates. Based on the studies found that the use as stabilizers structure oxygen smoothie dietary fibres «Citri-Fi» in concentrations of 0.8 – 1 % and non-starch polysaccharides at concentrations of 0.1 - 0.3 % contributes to the production of foams drinks with a sufficiently high magnification. Increasing the dose of dietary fibres in making smoothies viscosity increases and weighting system, drink bad whipped, there is separation of the product into phases, the system becomes thermodynamically unstable. Production technology and recipes for new types of smoothies with dietary fibres. Preventive orientation developed oxygenated drinks allows to include them in the diet of patients with chronic heart failure, which is confirmed by appropriate investigations.

Inducible expression of Bs2 R gene from Capsicum chacoense in sweet orange (Citrus sinensis L. Osbeck) confers enhanced resistance to citrus canker disease.

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Sendín, Lorena Noelia; Orce, Ingrid Georgina; Gómez, Rocío Liliana; Enrique, Ramón; Grellet Bournonville, Carlos Froilán; Noguera, Aldo Sergio; Vojnov, Adrián Alberto; Marano, María Rosa; Castagnaro, Atilio Pedro; Filippone, María Paula

2017-04-01

Transgenic expression of the pepper Bs2 gene confers resistance to Xanthomonas campestris pv. vesicatoria (Xcv) pathogenic strains which contain the avrBs2 avirulence gene in susceptible pepper and tomato varieties. The avrBs2 gene is highly conserved among members of the Xanthomonas genus, and the avrBs2 of Xcv shares 96% homology with the avrBs2 of Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker disease. A previous study showed that the transient expression of pepper Bs2 in lemon leaves reduced canker formation and induced plant defence mechanisms. In this work, the effect of the stable expression of Bs2 gene on citrus canker resistance was evaluated in transgenic plants of Citrus sinensis cv. Pineapple. Interestingly, Agrobacterium-mediated transformation of epicotyls was unsuccessful when a constitutive promoter (2× CaMV 35S) was used in the plasmid construction, but seven transgenic lines were obtained with a genetic construction harbouring Bs2 under the control of a pathogen-inducible promoter, from glutathione S-transferase gene from potato. A reduction of disease symptoms of up to 70% was observed in transgenic lines expressing Bs2 with respect to non-transformed control plants. This reduction was directly dependent on the Xcc avrBs2 gene since no effect was observed when a mutant strain of Xcc with a disruption in avrBs2 gene was used for inoculations. Additionally, a canker symptom reduction was correlated with levels of the Bs2 expression in transgenic plants, as assessed by real-time qPCR, and accompanied by the production of reactive oxygen species. These results indicate that the pepper Bs2 resistance gene is also functional in a family other than the Solanaceae, and could be considered for canker control.

DNA polymorphisms and biocontrol of Bacillus antagonistic to citrus bacterial canker with indication of the interference of phyllosphere biofilms.

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Tzu-Pi Huang

Full Text Available Citrus bacterial canker caused by Xanthomonas axonopodis pv. citri is a devastating disease resulting in significant crop losses in various citrus cultivars worldwide. A biocontrol agent has not been recommended for this disease. To explore the potential of bacilli native to Taiwan to control this disease, Bacillus species with a broad spectrum of antagonistic activity against various phytopathogens were isolated from plant potting mixes, organic compost and the rhizosphere soil. Seven strains TKS1-1, OF3-16, SP4-17, HSP1, WG6-14, TLB7-7, and WP8-12 showing superior antagonistic activity were chosen for biopesticide development. The genetic identity based on 16S rDNA sequences indicated that all seven native strains were close relatives of the B. subtilis group and appeared to be discrete from the B. cereus group. DNA polymorphisms in strains WG6-14, SP4-17, TKS1-1, and WP8-12, as revealed by repetitive sequence-based PCR with the BOXA1R primers were similar to each other, but different from those of the respective Bacillus type strains. However, molecular typing of the strains using either tDNA-intergenic spacer regions or 16S-23S intergenic transcribed spacer regions was unable to differentiate the strains at the species level. Strains TKS1-1 and WG6-14 attenuated symptom development of citrus bacterial canker, which was found to be correlated with a reduction in colonization and biofilm formation by X. axonopodis pv. citri on leaf surfaces. The application of a Bacillus strain TKS1-1 endospore formulation to the leaf surfaces of citrus reduced the incidence of citrus bacterial canker and could prevent development of the disease.

Detection of mechanical and disease stresses in citrus plants by fluorescence spectroscopy

Science.gov (United States)

Belasque, J., Jr.; Gasparoto, M. C. G.; Marcassa, L. G.

2008-04-01

We have investigated the detection of mechanical and disease stresses in citrus plants (Citrus limonia [L.] Osbeck) using laser-induced fluorescence spectroscopy. Due to its economic importance we have chosen to investigate the citrus canker disease, which is caused by the Xanthomonas axonopodis pv. citri bacteria. Mechanical stress was also studied because it plays an important role in the plant's infection by such bacteria. A laser-induced fluorescence spectroscopy system, composed of a spectrometer and a 532 nm10 mW excitation laser was used to perform fluorescence spectroscopy. The ratio of two chlorophyll fluorescence bands allows us to detect and discriminate between mechanical and disease stresses. This ability to discriminate may have an important application in the field to detect citrus canker infected trees.

Effect of Gamma Radiation on Chemical Composition and Antimicrobial Activity of Fennel and Geranium Volatile Oils

International Nuclear Information System (INIS)

Abo-Seoud, M. A.; Helal, I. M. M.; Sarhan, M. M.; Galal, A. M. M.

2004-01-01

Essential oils of fennel and geranium were gamma irradiated with doses of 0,10,20,30 and 40 kGy. The studied oils were tested for their antimicrobial activities against some pathogenic microorganisms (Alternaria alternata, Aspergillus niger, A. flavus, Fusarium oxysporium, Trichoderma viride and Pseudomonas citri). Both oils were used in four concentrations of 500, 1000, 2000 and 4000 ppm. The investigated oils showed different inhibition effects against the tested microorganisms. Gamma irradiated oils increased the antimicrobial activity with different magnitudes. Generally, increasing oil concentration increased antimicrobial activity of the used oils and that of 4000 ppm was the most effective one. The essential oils were analyzed by G.C. to evaluate the effect of gamma irradiation on the oil components. (authors)

Genome editing of the disease susceptibility gene CsLOB1 in citrus confers resistance to citrus canker.

Science.gov (United States)

Jia, Hongge; Zhang, Yunzeng; Orbović, Vladimir; Xu, Jin; White, Frank F; Jones, Jeffrey B; Wang, Nian

2017-07-01

Citrus is a highly valued tree crop worldwide, while, at the same time, citrus production faces many biotic challenges, including bacterial canker and Huanglongbing (HLB). Breeding for disease-resistant varieties is the most efficient and sustainable approach to control plant diseases. Traditional breeding of citrus varieties is challenging due to multiple limitations, including polyploidy, polyembryony, extended juvenility and long crossing cycles. Targeted genome editing technology has the potential to shorten varietal development for some traits, including disease resistance. Here, we used CRISPR/Cas9/sgRNA technology to modify the canker susceptibility gene CsLOB1 in Duncan grapefruit. Six independent lines, D LOB 2, D LOB 3, D LOB 9, D LOB 10, D LOB 11 and D LOB 12, were generated. Targeted next-generation sequencing of the six lines showed the mutation rate was 31.58%, 23.80%, 89.36%, 88.79%, 46.91% and 51.12% for D LOB 2, D LOB 3, D LOB 9, D LOB 10, D LOB 11 and D LOB 12, respectively, of the cells in each line. D LOB 2 and D LOB 3 showed canker symptoms similar to wild-type grapefruit, when inoculated with the pathogen Xanthomonas citri subsp. citri (Xcc). No canker symptoms were observed on D LOB 9, D LOB 10, D LOB 11 and D LOB 12 at 4 days postinoculation (DPI) with Xcc. Pustules caused by Xcc were observed on D LOB 9, D LOB 10, D LOB 11 and D LOB 12 in later stages, which were much reduced compared to that on wild-type grapefruit. The pustules on D LOB 9 and D LOB 10 did not develop into typical canker symptoms. No side effects and off-target mutations were detected in the mutated plants. This study indicates that genome editing using CRISPR technology will provide a promising pathway to generate disease-resistant citrus varieties. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

The post-transcriptional regulator rsmA/csrA activates T3SS by stabilizing the 5' UTR of hrpG, the master regulator of hrp/hrc genes, in Xanthomonas.

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Maxuel O Andrade

2014-02-01

Full Text Available The RsmA/CsrA family of the post-transcriptional regulators of bacteria is involved in the regulation of many cellular processes, including pathogenesis. In this study, we demonstrated that rsmA not only is required for the full virulence of the phytopathogenic bacterium Xanthomonas citri subsp. citri (XCC but also contributes to triggering the hypersensitive response (HR in non-host plants. Deletion of rsmA resulted in significantly reduced virulence in the host plant sweet orange and a delayed and weakened HR in the non-host plant Nicotiana benthamiana. Microarray, quantitative reverse-transcription PCR, western-blotting, and GUS assays indicated that RsmA regulates the expression of the type 3 secretion system (T3SS at both transcriptional and post-transcriptional levels. The regulation of T3SS by RsmA is a universal phenomenon in T3SS-containing bacteria, but the specific mechanism seems to depend on the interaction between a particular bacterium and its hosts. For Xanthomonads, the mechanism by which RsmA activates T3SS remains unknown. Here, we show that RsmA activates the expression of T3SS-encoding hrp/hrc genes by directly binding to the 5' untranslated region (UTR of hrpG, the master regulator of the hrp/hrc genes in XCC. RsmA stabilizes hrpG mRNA, leading to increased accumulation of HrpG proteins and subsequently, the activation of hrp/hrc genes. The activation of the hrp/hrc genes by RsmA via HrpG was further supported by the observation that ectopic overexpression of hrpG in an rsmA mutant restored its ability to cause disease in host plants and trigger HR in non-host plants. RsmA also stabilizes the transcripts of another T3SS-associated hrpD operon by directly binding to the 5' UTR region. Taken together, these data revealed that RsmA primarily activates T3SS by acting as a positive regulator of hrpG and that this regulation is critical to the pathogenicity of XCC.

Especificidade alimentar: em busca de um caráter taxonômico para a diferenciação de duas espécies crípticas de cochonilhas do gênero Planococcus (Hemíptera: Pseudococcidae

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Ana Luiza Viana de Sousa

2012-09-01

Full Text Available O objetivo deste trabalho foi detectar uma possível especificidade alimentar de duas espécies crípticas de cochonilhas do gênero Planococcus, refletida em seu desenvolvimento em frutos de cacaueiro, cafeeiro e citros. A cochonilha Planococcus minor (Maskell, obtida de frutos de cacau (Theobroma cacao L. cv. Comum, e Planococcus citri (Risso, de lavoura de café (Coffea arabica L. cv. Mundo Novo e de mudas de citros (Citrus sinensis L. Osbeck cv. Bahia, foram criadas em abóboras (Cucurbita maxima L., em laboratório. Rosetas com frutos de café foram mantidas sobre uma lâmina de 5 mm de ágar-água em placas de Petri, vedadas com filme plástico de polietileno. Em frutos de citros e cacau foram utilizadas gaiolas plásticas cilíndricas (1,5 cm x 3,0 cm, vedadas com voile na parte superior, as quais foram fixadas nos frutos por meio de um elástico. Os bioensaios foram conduzidos em câmaras climatizadas a 25 ± 1ºC, 70 ± 10% UR e 12 horas de fotofase, utilizando-se de 30 repetições. Para as cochonilhas provenientes de cafeeiro, o substrato café proporcionou o maior período ninfal de fêmeas (20,8 dias e maior longevidade (31,7 dias. Para fêmeas oriundas de cacau, o substrato cacau promoveu o menor período ninfal (21,1 dias e maior longevidade (25,0 dias. Para aquelas oriundas de citros, o substrato cacau promoveu o menor período ninfal (18,4 dias, e o substrato citros, a maior longevidade (32,0 dias. As maiores porcentagens de mortalidade (50% foram obtidas das ninfas oriundas de frutos de cacau e citros, criadas em café e cacau, e as menores foram das ninfas oriundas de frutos de café, independentemente do substrato em que foram criadas. A cochonilha P. minor mostra uma associação mais estreita com o cacau e, eventualmente, café, em relação ao citros, o que explicaria sua maior ocorrência em cacau. No entanto, P. citri não evidencia nenhuma especificidade para os três substratos testados.

Nanofilms of hyaluronan/chitosan assembled layer-by-layer: An antibacterial surface for Xylella fastidiosa.

Science.gov (United States)

Hernández-Montelongo, Jacobo; Nascimento, Vicente F; Murillo, Duber; Taketa, Thiago B; Sahoo, Prasana; de Souza, Alessandra A; Beppu, Marisa M; Cotta, Monica A

2016-01-20

In this work, nanofilms of hyaluronan/chitosan (HA/CHI) assembled layer by layer were synthesized; their application as a potential antimicrobial material was demonstrated for the phytopathogen Xylella fastidiosa, a gram-negative bacterium, here used as a model. For the synthesis, the influence of pH and ionic strength of these natural polymer stem-solutions on final characteristics of the HA/CHI nanofilms was studied in detail. The antibacterial effect was evaluated using widefield fluorescence microscopy. These results were correlated with the chemical properties of the nanofilms, studied by FTIR and Raman spectroscopy, as well as with their morphology and surface properties characterized using SEM and AFM. The present findings can be extended to design and optimize HA/CHI nanofilms with enhanced antimicrobial behavior for other type of phytopathogenic gram-negative bacteria species, such as Xanthomonas citri, Xanthomas campestri and Ralstonia solanacearum. Copyright © 2015 Elsevier Ltd. All rights reserved.

No evidence demonstrating hepatotoxicity associated with hydroxycitric acid

Institute of Scientific and Technical Information of China (English)

Sidney J Stohs; Harry G Preuss; Sunny E Ohia; Gilbert R Kaats; Carl L Keen; Lonnie D Williams; George A Burdock

2009-01-01

Although a number of cases of hepatotoxicity are associated with the use of Hydroxycut weight management products,it has been alleged that their effects are primarily due to the presence of hydroxycitric acid (HCA,as Super CitriMax) in the formulations.However,while these products contain up to 20 different ingredients,some do not contain HCA.Case studies reported to date have not considered in depth the literature on the numerous animal and human studies that have been conducted on the safety and efficacy of HCA.No HCAassociated hepatotoxicity or treatment-related adverse effects have been reported in these studies,and thus it is premature to make the assumptions presented in the recent case studies regarding Hydroxycut.If it is established in well controlled studies that the use of these formulations with and/or without HCA can result in the occurrence or progression of hepatotoxicity,additional studies should be conducted to characterize the causative factor(s).

Bacterial diversity analysis of Huanglongbing pathogen-infected citrus, using PhyloChip and 16S rRNA gene clone library sequencing

Energy Technology Data Exchange (ETDEWEB)

Shankar Sagaram, U.; DeAngelis, K.M.; Trivedi, P.; Andersen, G.L.; Lu, S.-E.; Wang, N.

2009-03-01

host plants (5,27,40). Tatineni and colleagues discovered that the HLB bacteria were unevenly distributed in phloem of bark tissue, vascular tissue of the leaf midrib, roots, and different floral and fruit parts (43). Unsuccessful attempts in culturing the pathogen are notably hampering efforts to understand its biology and pathogenesis mechanism. Using a modified Koch's Postulates approach, Jagoueix and colleagues were able to re-infect periwinkle plants from a mixed microbial community harvested from HLB diseased plants (25). Emergence of the disease in otherwise healthy plants led to the conclusion that HLB was associated with Candidatus Liberibacter sp. based on its 16S rDNA sequence (18,25). Currently, three species of the pathogen are recognized from trees with HLB disease based on 16S rDNA sequence: Ca. Liberibacter asiaticus (Las), Ca. Liberibacter africanus (Laf), and Ca. Liberibacter americanus (Lam); Las is the most prevalent species among HLB diseased trees (5,12,18,25,44). Las is naturally transmitted to citrus by the psyllid, Diaphorina citri Kuwayama, and can be artificially transmitted by grafting from citrus to citrus and dodder (Cuscuta campestris) to periwinkle (Catharanthus roseus) or tobacco (Nicotiana tabacum Xanthi) (5). Based on current research regarding the associations of Liberibacter in planta there is not enough evidence to implicate Liberibacter as the definitive causal agent of HLB disease due to its resistance to cultivation in vitro. It is possible that HLB disease may be the result of complex etiology where Liberibacter interacts with other endophytic bacteria. However, there is not enough evidence regarding its association(s) in planta to make this conclusion, nor is it known whether associated microbial communities play a role in expression of pathogenic traits. The main objective of the study was to test the hypothesis that other bacteria besides Ca. Liberibacter spp. are associated with citrus greening disease. The differences

Antalya İlinde Portakal Bahçelerinde Gözlenen Önemli Zararlı ve Yararlı Böcek Popülasyonları

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Vildan GÖL

2016-06-01

Full Text Available Bu çalışmada, 2014 yılında, Antalya ilinde bulunan iki portakal bahçesinde önemli zararlı ve yararlı böcek popülasyonları belirlenmiştir. Bahçelerde ağırlıklı olarak portakal bulunması nedeniyle örneklemeler de bu tür üzerinde yapılmıştır. Çalışma süresince zararlılara karşı herhangi bir mücadele yöntemi uygulanmamıştır. Çalışmada örneklerin toplanması için gözle kontrol yöntemi, yaprak, sürgün, meyve örnekleme yöntemi, darbe ve tuzakla yakalama yöntemi kullanılmıştır. Çalışma sonucunda zararlı türler olarak Tetranychus urticae (Koch (Acarina: Tetranychidae, Empoasca decipiens (Paoli, Asymmetrasca decedens (Paoli (Hemiptera: Cicadellidae, Dialeurodes citri (Ashmead, Aleurothrixus floccosus (Maskell (Hem.: Aleyrodidae, Aphis craccivora (Koch (Hem.: Aphididae, Icerya purchasi Maskell (Hem.: Margarodidae, Coccus hesperidum (Linnaeus, Ceroplastes floridensis (Comstock (Hem.: Coccidae, Aonidiella aurantii (Maskell, Aonidiella citrina (Coquillet, Parlatoria pergandii (Comstock (Hem.: Diaspididae, Planococcus citri (Risso (Hem.: Pseudococcidae, Ceratitis capitata (Wiedemann (Diptera: Tephritidae ve Phyllocnistis citrella (Stainton (Lepidoptera: Gracillaridae saptanmıştır. Bunlardan E. decipiens (Paoli, A. decedens (Paoli, A. floccosus (Maskell, A. aurantii (Maskell ve Parlatoria pergandii (Comstock en yaygın türler olarak belirlenmiştir. Ayrıca 2 takıma bağlı 7 avcı ve asalak tür saptanmıştır. Bu türler: Chrysoperla carnea (Stephens (Neuroptera: Chrysopidae, Coccinella septempunctata L., Chilocorus bipustulatus L., Rhyzobius lophantae (Blaisdell, Oenopia conglobata (L. (Coleoptera: Coccinellidae, Aphytis melinus DeBach ve Comperiella bifasciata (Coleoptera: Aphelinidae’ dır. Çalışmada ayrıca zararlı iki cicadellid türü ile A. floccosus, A. aurantii, P. pergandii, C. capitata‘nın ve doğal düşmanlardan C. carnea’nın, coccinellidlerin, A. melinus ve E. citrina�

Eficiência e custo do controle químico da mancha de alternaria em tangor murcote Efficiency and cost of chemical control of alternaria brown spot

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Adimara Bentivoglio Colturato

2009-09-01

Full Text Available A mancha de alternaria, causada por Alternaria alternata f. sp. citri, afeta tangelos Minneola, tangerinas Dancy, tangores Murcote e, menos freqüentemente, tangelos Orlando, tangerinas Novas, Lees e Sunburst. Esta doença causa desfolha grave, queda de frutos e manchas nas frutas. O objetivo deste trabalho foi estabelecer o melhor fungicida e a melhor dose para o controle da mancha marrom de alternaria. O delineamento experimental foi de parcelas subdivididas em blocos, com 10 tratamentos principais e 3 doses (subparcelas, com 5 repetições. Foram feitas 5 aplicações, com intervalo de 15 dias. Os tratamentos foram: azoxystrobin, pyraclostrobin, trifloxystrobin, trifloxystrobin + tebuconazole (2 aplicações seguido de 3 aplicações de mancozeb, difenoconazole, trifloxystrobin + propiconazole, iprodione, trifloxystrobin + propineb (2 aplicações seguido de 3 aplicações de oxicloreto de cobre, oxicloreto de cobre + óleo e testemunha. Simultaneamente foram feitas avaliações de incidência e número de lesões por folha. Ao surgimento dos frutos foram avaliadas a incidência em frutos e a produtividade em Kg/ha. Todos os tratamentos foram superiores à testemunha quanto a produtividade. Entre os produtos utilizados o tratamento com trifloxystrobin + propiconazole foi rentável comparando-se custo e produtividade.Alternaria brown spot, caused by Alternaria alternata sp. citri, attacks with more intensity the Tangelos Minneola, tangerine Dancy, and Murcotts, and with less intensity the tangelos Orlando and the tangerinas Novas, Lees and Sunburst. This disease causes severe defoliation and drop or necrotic spots in the fruits. The aim of this work was to evaluated the chemical control of brown spot, and to define the most appropriate dosage of fungicide to control it. The experimental design was split-spot, with ten treatments and 3 doses of fungicides, with five replicates, the fungicides were: azoxystrobin, pyraclostrobin, trifloxystrobin

Molecular evidence for ongoing complementarity and horizontal gene transfer in endosymbiotic systems of mealybugs

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Sergio eLópez-Madrigal

2014-08-01

Full Text Available Intracellular bacterial supply of essential amino acids is common among sap-feeding insects, thus complementing the scarcity of nitrogenous compounds in plant phloem. This is also the role of the two mealybug endosymbiotic systems whose genomes have been sequenced. In the nested endosymbiotic system from Planococcus citri (Pseudococcinae, Candidatus Tremblaya princeps and Candidatus Moranella endobia cooperate to synthesize essential amino acids, while in Phenacoccus avenae (Phenacoccinae this function is performed by its single endosymbiont Candidatus Tremblaya phenacola. However, little is known regarding the evolution of essential amino acid supplementation strategies in other mealybug systems. To address this knowledge gap, we screened for the presence of six selected loci involved in essential amino acid biosynthesis in five additional mealybug species. We found evidence of ongoing complementarity among endosymbionts from insects of subfamily Pseudococcinae, as well as horizontal gene transfer affecting endosymbionts from insects of family Phenacoccinae, providing a more comprehensive picture of the evolutionary history of these endosymbiotic systems. Additionally, we report two diagnostic motifs to help identify invasive mealybug species.

Permanent Genetic Resources added to Molecular Ecology Resources database 1 January 2009-30 April 2009.

Science.gov (United States)

Abercrombie, L G; Anderson, C M; Baldwin, B G; Bang, I C; Beldade, R; Bernardi, G; Boubou, A; Branca, A; Bretagnolle, F; Bruford, M W; Buonamici, A; Burnett, R K; Canal, D; Cárdenas, H; Caullet, C; Chen, S Y; Chun, Y J; Cossu, C; Crane, C F; Cros-Arteil, S; Cudney-Bueno, R; Danti, R; Dávila, J A; Della Rocca, G; Dobata, S; Dunkle, L D; Dupas, S; Faure, N; Ferrero, M E; Fumanal, B; Gigot, G; González, I; Goodwin, S B; Groth, D; Hardesty, B D; Hasegawa, E; Hoffman, E A; Hou, M L; Jamsari, A F J; Ji, H J; Johnson, D H; Joseph, L; Justy, F; Kang, E J; Kaufmann, B; Kim, K S; Kim, W J; Koehler, A V; Laitung, B; Latch, P; Liu, Y D; Manjerovic, M B; Martel, E; Metcalfe, S S; Miller, J N; Midgley, J J; Migeon, A; Moore, A J; Moore, W L; Morris, V R F; Navajas, M; Navia, D; Neel, M C; De Nova, P J G; Olivieri, I; Omura, T; Othman, A S; Oudot-Canaff, J; Panthee, D R; Parkinson, C L; Patimah, I; Pérez-Galindo, C A; Pettengill, J B; Pfautsch, S; Piola, F; Potti, J; Poulin, R; Raimondi, P T; Rinehart, T A; Ruzainah, A; Sarver, S K; Scheffler, B E; Schneider, A R R; Silvain, J F; Siti Azizah, M N; Springer, Y P; Stewart, C N; Sun, W; Tiedemann, R; Tsuji, K; Trigiano, R N; Vendramin, G G; Wadl, P A; Wang, L; Wang, X; Watanabe, K; Waterman, J M; Weisser, W W; Westcott, D A; Wiesner, K R; Xu, X F; Yaegashi, S; Yuan, J S

2009-09-01

This article documents the addition of 283 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Agalinis acuta; Ambrosia artemisiifolia; Berula erecta; Casuarius casuarius; Cercospora zeae-maydis; Chorthippus parallelus; Conyza canadensis; Cotesia sesamiae; Epinephelus acanthistius; Ficedula hypoleuca; Grindelia hirsutula; Guadua angustifolia; Leucadendron rubrum; Maritrema novaezealandensis; Meretrix meretrix; Nilaparvata lugens; Oxyeleotris marmoratus; Phoxinus neogaeus; Pristomyrmex punctatus; Pseudobagrus brevicorpus; Seiridium cardinale; Stenopsyche marmorata; Tetranychus evansi and Xerus inauris. These loci were cross-tested on the following species: Agalinis decemloba; Agalinis tenella; Agalinis obtusifolia; Agalinis setacea; Agalinis skinneriana; Cercospora zeina; Cercospora kikuchii; Cercospora sorghi; Mycosphaerella graminicola; Setosphaeria turcica; Magnaporthe oryzae; Cotesia flavipes; Cotesia marginiventris; Grindelia Xpaludosa; Grindelia chiloensis; Grindelia fastigiata; Grindelia lanceolata; Grindelia squarrosa; Leucadendron coniferum; Leucadendron salicifolium; Leucadendron tinctum; Leucadendron meridianum; Laodelphax striatellus; Sogatella furcifera; Phoxinus eos; Phoxinus rigidus; Phoxinus brevispinosus; Phoxinus bicolor; Tetranychus urticae; Tetranychus turkestani; Tetranychus ludeni; Tetranychus neocaledonicus; Tetranychus amicus; Amphitetranychus viennensis; Eotetranychus rubiphilus; Eotetranychus tiliarium; Oligonychus perseae; Panonychus citri; Bryobia rubrioculus; Schizonobia bundi; Petrobia harti; Xerus princeps; Spermophilus tridecemlineatus and Sciurus carolinensis. © 2009 Blackwell Publishing Ltd.

Litter decomposing fungi in sal (Shorea robusta forests of central India

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RAM KEERTI VERMA

2011-11-01

Full Text Available Soni KK, Pyasi A, Verma RK. 2011. Litter decomposing fungi in sal (Shorea robusta forests of central India. Nusantara Bioscience 3: 136-144. The present study aim on isolation and identification of fungi associated with decomposition of litter of sal forest in central India. Season wise successional changes in litter mycoflora were determined for four main seasons of the year namely, March-May, June-August, September-November and December-February. Fungi like Aspergillus flavus, A. niger and Rhizopus stolonifer were associated with litter decomposition throughout the year, while Aspergillus fumigatus, Cladosporium cladosporioides, C. oxysporum, Curvularia indica, and C. lunata were recorded in three seasons. Some fungi including ectomycorrhiza forming occur only in the rainy season (June-August these are Astraeus hygrometricus, Boletus fallax, Calvatia elata, Colletotrichum dematium, Corticium rolfsii, Mycena roseus, Periconia minutissima, Russula emetica, Scleroderma bovista, S. geaster, S. verrucosum, Scopulariopsis alba and four sterile fungi. Fungi like Alternaria citri, Gleocladium virens, Helicosporium phragmitis and Pithomyces cortarum were rarely recorded only in one season.

Crystal Structure and Regulation of the Citrus Pol III Repressor MAF1 by Auxin and Phosphorylation.

Science.gov (United States)

Soprano, Adriana Santos; Giuseppe, Priscila Oliveira de; Shimo, Hugo Massayoshi; Lima, Tatiani Brenelli; Batista, Fernanda Aparecida Heleno; Righetto, Germanna Lima; Pereira, José Geraldo de Carvalho; Granato, Daniela Campos; Nascimento, Andrey Fabricio Ziem; Gozzo, Fabio Cesar; de Oliveira, Paulo Sérgio Lopes; Figueira, Ana Carolina Migliorini; Smetana, Juliana Helena Costa; Paes Leme, Adriana Franco; Murakami, Mario Tyago; Benedetti, Celso Eduardo

2017-09-05

MAF1 is the main RNA polymerase (Pol) III repressor that controls cell growth in eukaryotes. The Citrus ortholog, CsMAF1, was shown to restrict cell growth in citrus canker disease but its role in plant development and disease is still unclear. We solved the crystal structure of the globular core of CsMAF1, which reveals additional structural elements compared with the previously available structure of hMAF1, and explored the dynamics of its flexible regions not present in the structure. CsMAF1 accumulated in the nucleolus upon leaf excision, and this translocation was inhibited by auxin and by mutation of the PKA phosphorylation site, S45, to aspartate. Additionally, mTOR phosphorylated recombinant CsMAF1 and the mTOR inhibitor AZD8055 blocked canker formation in normal but not CsMAF1-silenced plants. These results indicate that the role of TOR on cell growth induced by Xanthomonas citri depends on CsMAF1 and that auxin controls CsMAF1 interaction with Pol III in citrus. Copyright © 2017 Elsevier Ltd. All rights reserved.

Differential effects of exposure to ambient vanilla and citris aromas on mood, arousal and food choice

NARCIS (Netherlands)

Wijk, de R.A.; Zijlstra, S.

2012-01-01

Background Aromas have been associated with physiological, psychological affective and behavioral effects. We tested whether effects of low-level exposure to two ambient food-related aromas (citrus and vanilla) could be measured with small numbers of subjects, low-cost physiological sensors and

Effect Of GAMMA Radiation On Antimicrobial Activity And Chemical Constituents Of Marjoram (Majorana Hortensis Essential Oil

International Nuclear Information System (INIS)

GALAL, Y.G.M.; EL-GHANDOUR, I.A.; ABOU SEER, A.M.M.; DESOUKY, E.M.; ARAFA, R.A.

2010-01-01

Field experiment was set up to study the response of marjoram plant grown on sandy soil and inoculated with Bradyrhizobium sp. and/or B. polymixa in combination with organic fertilizers. The extracted oil was irradiated with gamma irradiation at doses of 10, 20, 30, 40 and 50 kGy. The chemical constituents of the essential oils were analyzed by GC-Ms techniques after exposure to gamma radiation. The gamma irradiated essential oils of marjoram were tested for their antimicrobial activities against some pathogenic microorganisms i.e. Klebsiella pneumoniae, Bacillus cereus, Staphylococcus aureus, Salmonella enteritidis, Pseudomonas citri, Fusarium oxysporum, Aspergillus niger, Trichoderma viride and Aspergillus flavus. Extracted oil was the best when soil was treated with faba bean straw and inoculated with B. polymixa + Bradyrhizobium sp. Similarly, the extracted oil from plant cultivated in soil treated with sheep manure in combination with B. polymixa recorded the highest value. Results also revealed that gamma irradiation doses increased the antimicrobial activity with different magnitudes. The essential oil extracted from herb exposed to 30 kGy was found to be the most active antimicrobial with slight increases in the main components.

Some biological activities of pigments extracted from Micrococcus roseus (PTCC 1411) and Rhodotorula glutinis (PTCC 5257).

Science.gov (United States)

Rostami, Hossein; Hamedi, Hassan; Yolmeh, Mahmoud

2016-12-01

The importance of replacing synthetic pigments with natural types is increasing day by day in the food industry due to the harmful effects of some synthetic pigments. Microorganisms are a major source of natural pigments, which nowadays have attracted the attention of researchers. In this study, carotenoid pigments were produced by Micrococcus roseus and Rhodotorula glutinis, and some of their biological properties such as antimicrobial, antioxidant, anticancer, and anti-inflammatory activities were evaluated. Given the results, bacteria, especially gram-positive bacteria, had higher sensitivity to the pigments extracted from M. roseus (PEM) and R. glutinis (PER) compared to molds so that Bacillus cereus and Alternaria citri had the highest and the lowest sensitivity, respectively. PER showed a higher antioxidant activity compared with PEM in the various methods of measuring antioxidant activity. In vitro and in vivo anti-tumor-promoting activities of PER were measured significantly more than PEM (P <0.05). Both pigment extracts remarkably inhibited the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation, so that ID 50 (50% inhibitory dose) of PEM and PER were 0.22 and 0.09 mg/ear, respectively. © The Author(s) 2016.

Antimicrobial activities of pyridinium-tailored pyrazoles bearing 1,3,4-oxadiazole scaffolds

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Lei Zhou

2017-11-01

Full Text Available Herein, a series of pyridinium-tailored 5-trifluoromethylpyrazoles containing 1,3,4-oxadiazole moieties were constructed through coupling key pharmaceutical fragments of pyridinium, pyrazole, and 1,3,4-oxadiazole scaffolds in single molecular architecture. Antimicrobial results suggested that this kind of compounds exhibited significant activities against three types of pathogenic bacteria and six fungal strains in vitro. The minimal EC50 values of designed compounds against Xanthomonas oryzae pv. oryzae, Ralstonia solanacearum, and Xanthomonas axonopodis pv. citri could reach to 0.467, 1.04, and 0.600 μg/mL, respectively, through tuning and optimizing N-substituents, bridging atom, and alkyl length of the tailor. Antifungal assays revealed that all title molecules possessed considerable activity against Botrytis cinerea with the minimal EC50 value up to 2.71 μg/mL; and compounds I-8, I-10, I-12, II-12, and IV-12 showed the strongest growth suppression toward Rhizoctonia solani with EC50 values ranging from 10.2 to 24.0 μg/mL. Given the above results, this kind of compounds could serve as new lead compounds in the research of antimicrobial chemotherapy.

Application of droplet digital PCR for quantitative detection of Spiroplasma citri in comparison with real time PCR

Science.gov (United States)

Droplet digital Polymerase chain reaction (ddPCR) is a unique approach to measure the absolute copy number of nucleic acid targets without the need of external standards. It is a promising DNA quantification technology for medical diagnostics but there are only a few reports of its use for plant pat...

Multiple gene genealogies and phenotypic characters differentiate several novel species of Mycosphaerella and related anamorphs on banana.

Science.gov (United States)

Arzanlou, M; Groenewald, J Z; Fullerton, R A; Abeln, E C A; Carlier, J; Zapater, M-F; Buddenhagen, I W; Viljoen, A; Crous, P W

2008-06-01

Three species of Mycosphaerella, namely M. eumusae, M. fijiensis, and M. musicola are involved in the Sigatoka disease complex of bananas. Besides these three primary pathogens, several additional species of Mycosphaerella or their anamorphs have been described from Musa. However, very little is known about these taxa, and for the majority of these species no culture or DNA is available for study. In the present study, we collected a global set of Mycosphaerella strains from banana, and compared them by means of morphology and a multi-gene nucleotide sequence data set. The phylogeny inferred from the ITS region and the combined data set containing partial gene sequences of the actin gene, the small subunit mitochondrial ribosomal DNA and the histone H3 gene revealed a rich diversity of Mycosphaerella species on Musa. Integration of morphological and molecular data sets confirmed more than 20 species of Mycosphaerella (incl. anamorphs) to occur on banana. This study reconfirmed the previously described presence of Cercospora apii, M. citri and M. thailandica, and also identified Mycosphaerella communis, M. lateralis and Passalora loranthi on this host. Moreover, eight new species identified from Musa are described, namely Dissoconium musae, Mycosphaerella mozambica, Pseudocercospora assamensis, P. indonesiana, P. longispora, Stenella musae, S. musicola, and S. queenslandica.

Shedding light on the role of photosynthesis in pathogen colonization and host defense

KAUST Repository

Garavaglia, Betiana S.; Thomas, Ludivine; Gottig, Natalia; Zimaro, Tamara; Garofalo, Cecilia G.; Gehring, Christoph A; Ottado, Jorgelina

2010-01-01

The role of photosynthesis in plant defense is a fundamental question awaiting further molecular and physiological elucidation. To this end we investigated host responses to infection with the bacterial pathogen Xanthomonas axonopodis pv. citri, the pathogen responsible for citrus canker. This pathogen encodes a plant-like natriuretic peptide (XacPNP) that is expressed specifically during the infection process and prevents deterioration of the physiological condition of the infected tissue. Proteomic assays of citrus leaves infected with a XacPNP deletion mutant (DeltaXacPNP) resulted in a major reduction in photosynthetic proteins such as Rubisco, Rubisco activase and ATP synthase as a compared with infection with wild type bacteria. In contrast, infiltration of citrus leaves with recombinant XacPNP caused an increase in these host proteins and a concomitant increase in photosynthetic efficiency as measured by chlorophyll fluorescence assays. Reversion of the reduction in photosynthetic efficiency in citrus leaves infected with DeltaXacPNP was achieved by the application of XacPNP or Citrus sinensis PNP lending support to a case of molecular mimicry. Finally, given that DeltaXacPNP infection is less successful than infection with the wild type, it appears that reducing photosynthesis is an effective plant defense mechanism against biotrophic pathogens.

Genetic diversity of citrus bacterial canker pathogens preserved in herbarium specimens.

Science.gov (United States)

Li, Wenbin; Song, Qijian; Brlansky, Ronald H; Hartung, John S

2007-11-20

Citrus bacterial canker (CBC) caused by Xanthomonas axonopodis pv. citri (Xac) was first documented in India and Java in the mid 19th century. Since that time, the known distribution of the disease has steadily increased. Concurrent with the dispersion of the pathogen, the diversity of described strains continues to increase, with novel strains appearing in Saudi Arabia, Iran, and Florida in the last decade. Herbarium specimens of infected plants provide an historical record documenting both the geographic distribution and genetic diversity of the pathogen in the past. However, no method was available to assess the genetic diversity within these herbarium samples. We have developed a method, insertion event scanning (IES), and applied the method to characterize the diversity present within CBC populations documented as herbarium specimens over the past century. IES is based on the specific amplification of junction fragments that define insertion events. The potential for IES in current forensic applications is demonstrated by finding an exact match of pathogen genotypes preserved in herbarium specimens from Japan and Florida, demonstrating the source of the original outbreak of citrus canker in Florida in 1911. IES is a very sensitive technique for differentiating bacterial strains and can be applied to any of the several hundred bacteria for which full genomic sequence data are available.

Suspension of Egg Hatching Caused by High Humidity and Submergence in Spider Mites.

Science.gov (United States)

Ubara, Masashi; Osakabe, Masahiro

2015-08-01

We tested the effects of high humidity and submergence on egg hatching of spider mites. In both the high humidity and submergence treatments, many Tetranychus and Panonychus eggs did not hatch until after the hatching peak of the lower humidity or unsubmerged controls. However, after humidity decreased or water was drained, many eggs hatched within 1-3 h. This was observed regardless of when high humidity or submergence treatments were implemented: either immediately after oviposition or immediately before hatching was due. Normal eyespot formation was observed in most eggs in the high humidity and submergence treatments, which indicates that spider mite embryos develop even when eggs are underwater. Therefore, delays in hatching are not caused by delayed embryonic development. A delay in hatching was always observed in Panonychus citri (McGregor) but was more variable in Tetranychus urticae Koch and Tetranychus kanzawai Kishida. The high humidity and submergence treatments affected but did not suppress larval development in these species. In contrast, many Oligonychus eggs died following the high humidity treatments. In Tetranychus and Panonychus spider mites, suspension of egg hatching may mitigate the adverse effects of rainfall. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Shedding light on the role of photosynthesis in pathogen colonization and host defense

KAUST Repository

Garavaglia, Betiana S.

2010-09-01

The role of photosynthesis in plant defense is a fundamental question awaiting further molecular and physiological elucidation. To this end we investigated host responses to infection with the bacterial pathogen Xanthomonas axonopodis pv. citri, the pathogen responsible for citrus canker. This pathogen encodes a plant-like natriuretic peptide (XacPNP) that is expressed specifically during the infection process and prevents deterioration of the physiological condition of the infected tissue. Proteomic assays of citrus leaves infected with a XacPNP deletion mutant (DeltaXacPNP) resulted in a major reduction in photosynthetic proteins such as Rubisco, Rubisco activase and ATP synthase as a compared with infection with wild type bacteria. In contrast, infiltration of citrus leaves with recombinant XacPNP caused an increase in these host proteins and a concomitant increase in photosynthetic efficiency as measured by chlorophyll fluorescence assays. Reversion of the reduction in photosynthetic efficiency in citrus leaves infected with DeltaXacPNP was achieved by the application of XacPNP or Citrus sinensis PNP lending support to a case of molecular mimicry. Finally, given that DeltaXacPNP infection is less successful than infection with the wild type, it appears that reducing photosynthesis is an effective plant defense mechanism against biotrophic pathogens.

Evaluation of Antimicrobial Activity of Ginger (Zingiber officinale) Essential Oil Treated by Gamma Radiation

International Nuclear Information System (INIS)

Abdeldaiem, M.H.; Helal, I.M.

2009-01-01

The essential oil of ginger (Zingiber officinale) was irradiated at doses of 0,5, 10 and 15 kGy. The irradiated as well as non-irradiated oil were tested for their antibacterial activity against one strain of food poisoning bacteria (Bacillus cereus), one strain of the indicator microorganisms (Escherichia coli), two strains of human pathogenic bacteria (Klebsiella pneumonia, and Pseudomonas aeruginosa), one strain of food spoilage bacteria (Bacillus subtilis) and one strain of plant pathogenic bacteria(Pseudomonas citri). In addition, the studied oils were tested for their antifungal activities against five strains of plant pathogenic fungi (Aspergillus niger, Aspergillus flavus, Fusarium oxysporium, Trichoderma viride and Alternaria alternata) to investigate the effect of gamma irradiation on the antimicrobial activity of the tested oil. The obtained results showed that all tested oils exhibited an inhibition effect for the growth of the microorganisms under investigation. While, Trichoderma viride appeared to be the most resistant fungus with non-irradiated and irradiated ginger essential oil treatments. The ginger essential oil irradiated at dose of 10 kGy appeared to be the best one for inhibition the growth of most tested bacteria and fungi under investigation. Also, changes in chemical constituents of ginger essential oil as affected by gamma radiation were investigated by Gas Chromatography

A Phosphorylation Switch on Lon Protease Regulates Bacterial Type III Secretion System in Host

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Xiaofeng Zhou

2018-01-01

Full Text Available Most pathogenic bacteria deliver virulence factors into host cytosol through type III secretion systems (T3SS to perturb host immune responses. The expression of T3SS is often repressed in rich medium but is specifically induced in the host environment. The molecular mechanisms underlying host-specific induction of T3SS expression is not completely understood. Here we demonstrate in Xanthomonas citri that host-induced phosphorylation of the ATP-dependent protease Lon stabilizes HrpG, the master regulator of T3SS, conferring bacterial virulence. Ser/Thr/Tyr phosphoproteome analysis revealed that phosphorylation of Lon at serine 654 occurs in the citrus host. In rich medium, Lon represses T3SS by degradation of HrpG via recognition of its N terminus. Genetic and biochemical data indicate that phosphorylation at serine 654 deactivates Lon proteolytic activity and attenuates HrpG proteolysis. Substitution of alanine for Lon serine 654 resulted in repression of T3SS gene expression in the citrus host through robust degradation of HrpG and reduced bacterial virulence. Our work reveals a novel mechanism for distinct regulation of bacterial T3SS in different environments. Additionally, our data provide new insight into the role of protein posttranslational modification in the regulation of bacterial virulence.

Produção de frutos e incidência de cancro cítrico em laranjeiras "Monte Parnaso" enxertadas sobre sete porta-enxertos Fruit production and incidence of citrus canker lesions on "Monte Parnaso" navel orange budded on seven rootstocks

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Bernadete Reis

2008-06-01

Full Text Available Porta-enxertos de citros podem exercer influência sobre a produção de frutos e suscetibilidade das plantas ao cancro cítrico. Este trabalho objetivou selecionar porta-enxertos que induzam elevado índice de produtividade e menor suscetibilidade das copas de laranjeiras-de-umbigo "Monte Parnaso"(Citrus sinensis à bactéria Xanthomonas axonopodis pv. citri, causadora do cancro cítrico (CC. O experimento foi desenvolvido em um pomar experimental com 10 anos de idade, plantado em espaçamento de 2,5 x 6,0m, no município de Butiá, no Estado do Rio Grande do Sul. O delineamento experimental foi em blocos ao acaso, com sete tratamentos e quatro repetições, usando duas plantas por parcela. Os tratamentos constaram dos seguintes porta-enxertos: limoeiros "Cravo" (C. limonia e "Volkameriano" (C. volkameriana., trifoliata "Flying Dragon" (Poncirus trifoliata., laranjeira "Caipira" (C. sinensis, citrangeiro "Troyer" (C. sinensis x P. trifoliata, tangerineira "Sunki" (C. sunki. e citrumeleiro "Swingle" (C. paradisi x P. trifoliata. Os parâmetros avaliados foram: incidência de CC em folhas e frutos em toda a planta e em ramos previamente selecionados, em diversas épocas. Verificou-se que as plantas enxertadas sobre citrumeleiro "Swingle" apresentaram o maior índice de produtividade e, à semelhança do "Flying Dragon", a menor incidência de CC, tanto nas folhas como nos frutos. Além disso, verificou-se também que as plantas enxertadas sobre os limoeiros "Cravo" e "Volkameriana", apesar de produzirem as mais elevadas cargas de frutos, foram as mais suscetíveis ao CC.Citrus rootstocks can exert some influences on fruit production and susceptibility of the plants to citrus canker (CC. This research was performed aiming to select rootstocks which induce higher productivity index and lower susceptibility of 'Monte Parnaso' navel orange (Citrus sinensis trees to the bacteria Xanthomonas axonopodis pv. citri. The research was carried out in an

Volatile antimicrobials from Muscodor crispans, a novel endophytic fungus.

Science.gov (United States)

Mitchell, Angela M; Strobel, Gary A; Moore, Emily; Robison, Richard; Sears, Joe

2010-01-01

Muscodor crispans is a recently described novel endophytic fungus of Ananas ananassoides (wild pineapple) growing in the Bolivian Amazon Basin. The fungus produces a mixture of volatile organic compounds (VOCs); some of the major components of this mixture, as determined by GC/MS, are propanoic acid, 2-methyl-, methyl ester; propanoic acid, 2-methyl-; 1-butanol, 3-methyl-;1-butanol, 3-methyl-, acetate; propanoic acid, 2-methyl-, 2-methylbutyl ester; and ethanol. The fungus does not, however, produce naphthalene or azulene derivatives as has been observed with many other members of the genus Muscodor. The mixture of VOCs produced by M. crispans cultures possesses antibiotic properties, as does an artificial mixture of a majority of the components. The VOCs of the fungus are effective against a wide range of plant pathogens, including the fungi Pythium ultimum, Phytophthora cinnamomi, Sclerotinia sclerotiorum and Mycosphaerella fijiensis (the black sigatoka pathogen of bananas), and the serious bacterial pathogen of citrus, Xanthomonas axonopodis pv. citri. In addition, the VOCs of M. crispans killed several human pathogens, including Yersinia pestis, Mycobacterium tuberculosis and Staphylococcus aureus. Artificial mixtures of the fungal VOCs were both inhibitory and lethal to a number of human and plant pathogens, including three drug-resistant strains of Mycobacterium tuberculosis. The gaseous products of Muscodor crispans potentially could prove to be beneficial in the fields of medicine, agriculture, and industry.

Prunus cerasifera Ehrh. fabricated ZnO nano falcates and its photocatalytic and dose dependent in vitro bio-activity

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Jaffri Shaan Bibi

2018-03-01

Full Text Available Zinc oxide nano falcates of sickle shape have been synthesized from Prunus cerasifera pomological extract as a reducing cum stabilizing agent via novel, biomimetic and non-toxic route. Zinc oxide nano falcates were analyzed via ultraviolet spectroscopy, Fourier transform infrared analysis, X-ray powder diffraction, scanning electron microscopy and atomic force microscopy. Highly stable zinc oxide nano falcates synthesized at 200°C and 400°C calcination temperatures expressed intense UV-vis peak at 398 nm. Phenolic and amino groups were revealed by FTIR in pomological extract. Wurtzite crystalline structure of zinc oxide nano falcates was confirmed by XRD with average crystal size of 4.93 nm. SEM sizes ranged between 72.11-120 nm and 56.57-107.70 nm, respectively and shown higher polydispersity levels for two calcination temperatures. Augmented photocatalytic degradation of methyl red and bromophenol blue under direct solar irradiance shown pseudo first order kinetics (R2= 0.99 and 0.96. Furthermore, biomedical and agriculturally important pathogenic strains i.e., Xanthomanas axonopodis pv. citri and Pseudomonas syringae, Aspergillus niger, Aspergillus flavus, Aspergillus fumigatus, Aspergillus terreus, Penicillium chrysogenum, Fusarium solani and Lasiodiplodia theobromae were remarkably inhibited. Enhanced photocatalytic and antimicrobial activity reveals zinc oxide nano falcates promising prospects in nano bioremediation of polluted water and conversion into green nano pesticides.

Complete Sequence of p07-406, a 24,179-base-pair plasmid harboring the blaVIM-7 metallo-beta-lactamase gene in a Pseudomonas aeruginosa isolate from the United States.

Science.gov (United States)

Li, Hongyang; Toleman, Mark A; Bennett, Peter M; Jones, Ronald N; Walsh, Timothy R

2008-09-01

An outbreak involving a Pseudomonas aeruginosa strain that was resistant to all tested antimicrobials except polymyxin B occurred in a hospital in Houston, TX. Previous studies on this strain showed that it possesses a novel mobile metallo-beta-lactamase (MBL) gene, designated bla(VIM-7), located on a plasmid (p07-406). Here, we report the complete sequence, annotation, and functional characterization of this plasmid. p07-406 is 24,179 bp in length, and 29 open reading frames were identified related to known or putatively recognized proteins. Analysis of this plasmid showed it to be comprised of four distinct regions: (i) a region of 5,200 bp having a Tn501-like mercuric resistance (mer) transposon upstream of the replication region; (ii) a Tn3-like transposon carrying a truncated integron with a bla(VIM-7) gene and an insertion sequence inserted at the other end of this transposon; (iii) a region of four genes, upstream of the Tn3-like transposon, possessing very high similarity to plasmid pXcB from Xanthomonas campestris pv. citri commonly associated with plants; (iv) a backbone sequence similar to the backbone structure of the IncP group plasmid Rms149, pB10, and R751. This is the first plasmid to be sequenced carrying an MBL gene and highlights the amelioration of DNA segments from disparate origins, most noticeably from plant pathogens.

Effects of herbal mixture extracts on obesity in rats fed a high-fat diet

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Mei-Yin Chien

2016-07-01

Full Text Available The aim of this study was to investigate and compare the effects of three herbal mixture extracts on obesity induced by high-fat diet (HFD in rats. The prescriptions—Pericarpium citri reticulatae and Fructus crataegi—were used as matrix components and mixed with Ampelopsis grossedentata, Salvia miltiorrhiza, and epigallocatechin-3-gallate (EGCG to form T1, T2, and T3 complexes, respectively. Results revealed that HFD feeding significantly increased body weight gain, fat deposition, plasma lipid profiles, hepatic lipid accumulation, and hepatic vacuoles formation, but decreased plasma levels of adiponectin in rats. Only the T1 complex showed the tendency, although not significantly so, for decreased HFD-induced body weight gain. T1 and T3 complexes significantly reduced HFD-induced fat deposition, and plasma levels of triglyceride, total cholesterol, and low-density lipoprotein cholesterol. Only the T1 complex significantly increased HFD-reduced adiponectin levels in plasma, but decreased HFD-increased triglyceride content in liver tissues. All complexes effectively inhibited HFD-induced vacuoles formation. The content of dihydromyricetin, salvianolic acid B, and EGCG in T1, T2, and T3 complexes was 18.25 ± 0.07%, 22.20 ± 0.10%, and 18.86 ± 0.04%, respectively. In summary, we demonstrated that herbal mixture extracts, especially T1 complex, exhibit antiobesity activity in HFD-fed rats.

Effect of Gamma Irradiation on Chemical Constituents, Antimicrobials and Antioxidants of Thyme and Cinnamon Volatile Oils

International Nuclear Information System (INIS)

Anwar, M.M.; Nasr, E.H.; Ali, S.E.

2015-01-01

This study was carried out to investigate the effect of gamma irradiation at doses 10 or 20 kGy on the chemical constituents of essential oils extracted from irradiated thyme and cinnamon, and study the effect of these essential oils on the oxidative stability of soybean oil. In addition, this study was carried out to evaluate the efficacy of extracted essential oils from irradiated thyme and cinnamon as Antimicrobial agents.The results showed that the major components for essential oils of thyme were 77.03% thymol and 15.34 % eugenol acetate whereas the major components for essential oils of cinnamon were 66.14% cinnamaldehyde and 11.18% cinnamyl cinnamate. The study revealed that the essential oils extracted from irradiated cinnamon at 20 kGy showed the best antioxidant activities while gamma irradiation reduced the antioxidant activities of thyme essential oils. Essential oils extracted from irradiated and non-irradiated thyme and cinnamon were tested for the antibacterial activities against eight strains of Salmonella spp., Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas citri, Pseudomonas aeruginosa, Bacillus cereus and Bacillus subtilis and also tested for their antifungal activities against four strains of Penicillium spp., Rhizopus spp., Alternaria alternata and Aspergillus niger. The results showed that all tested oils exhibited an inhibition effect for the growth of the microorganisms under investigation and gamma irradiation at dose 20 kGy had the high effect.

Coffea arabica L., a new host plant for Acetobacter diazotrophicus, and isolation of other nitrogen-fixing acetobacteria.

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Jimenez-Salgado, T; Fuentes-Ramirez, L E; Tapia-Hernandez, A; Mascarua-Esparza, M A; Martinez-Romero, E; Caballero-Mellado, J

1997-09-01

Acetobacter diazotrophicus was isolated from coffee plant tissues and from rhizosphere soils. Isolation frequencies ranged from 15 to 40% and were dependent on soil pH. Attempts to isolate this bacterial species from coffee fruit, from inside vesicular-arbuscular mycorrhizal fungi spores, or from mealybugs (Planococcus citri) associated with coffee plants were not successful. Other acid-producing diazotrophic bacteria were recovered with frequencies of 20% from the coffee rhizosphere. These N2-fixing isolates had some features in common with the genus Acetobacter but should not be assigned to the species Acetobacter diazotrophicus because they differed from A. diazotrophicus in morphological and biochemical traits and were largely divergent in electrophoretic mobility patterns of metabolic enzymes at coefficients of genetic distance as high as 0.950. In addition, these N2-fixing acetobacteria differed in the small-subunit rRNA restriction fragment length polymorphism patterns obtained with EcoRI, and they exhibited very low DNA-DNA homology levels, ranging from 11 to 15% with the A. diazotrophicus reference strain PAI 5T. Thus, some of the diazotrophic acetobacteria recovered from the rhizosphere of coffee plants may be regarded as N2-fixing species of the genus Acetobacter other than A. diazotrophicus. Endophytic diazotrophic bacteria may be more prevalent than previously thought, and perhaps there are many more potentially beneficial N2-fixing bacteria which can be isolated from other agronomically important crops.

The economic importance of acaricides in the control of phytophagous mites and an update on recent acaricide mode of action research.

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Van Leeuwen, Thomas; Tirry, Luc; Yamamoto, Atsushi; Nauen, Ralf; Dermauw, Wannes

2015-06-01

Acaricides are one of the cornerstones of an efficient control program for phytophagous mites. An analysis of the global acaricide market reveals that spider mites such as Tetranychus urticae, Panonychus citri and Panonychus ulmi are by far the most economically important species, representing more than 80% of the market. Other relevant mite groups are false spider mites (mainly Brevipalpus), rust and gall mites and tarsonemid mites. Acaricides are most frequently used in vegetables and fruits (74% of the market), including grape vines and citrus. However, their use is increasing in major crops where spider mites are becoming more important, such as soybean, cotton and corn. As revealed by a detailed case study of the Japanese market, major shifts in acaricide use are partially driven by resistance development and the commercial availability of compounds with novel mode of action. The importance of the latter cannot be underestimated, although some compounds are successfully used for more than 30 years. A review of recent developments in mode of action research is presented, as such knowledge is important for devising resistance management programs. This includes spirocyclic keto-enols as inhibitors of acetyl-CoA carboxylase, the carbazate bifenazate as a mitochondrial complex III inhibitor, a novel class of complex II inhibitors, and the mite growth inhibitors hexythiazox, clofentezine and etoxazole that interact with chitin synthase I. Copyright © 2014 Elsevier Inc. All rights reserved.

Genetic similarity among Agistemus pallinii Matioli et al (Acari: Stigmaeidae) found in citrus orchards in Vicosa, Minas Gerais state, Brazil

International Nuclear Information System (INIS)

Matioli, Andre L.; Pallini, Angelo; Tavares, Mara G.

2009-01-01

Stigmaeidae are very important predators of mite and insect pests on several crops in Brazil. It is considered the second most important family of predatory mites in citrus orchards in Brazil. However, their identification, especially that of the members of the genus Agistemus, is rather difficult based only on morphology. Hence, this study describes the use of random amplified polymorphic DNA (RAPD-PCR) markers to determine the genetic similarity of an Agistemus pallinii Matioli et al population found in 2004 in a citrus orchard in Vicosa, Minas Gerais State, Brazil, preying on Panonychus citri (McGregor). Amplifications were performed with 12 random primers (OPAA8, OPAA19, OPAB1, OPAB5, OPAB18, OPAC9, OPAC17, OPAC19, OPAD10, OPAE9, OPAE12 and OPAE17), which generated 119 bands, with 53.8% polymorphism. The coefficients of genetic similarity among the individuals ranged from 0.68 to 0.99, indicating a high genetic similarity among them. The 3D projection analysis clustered the majority of individuals confirming their high similarity. Though individuals of A. pallinii are minute (± 360 μm long), the PCR-RAPD technique can still be used for their identification, complementing morphological analyses or for comparison of populations collected in different geographic regions. This is the fi rst molecular study carried out with stigmaeid mites. (author)

Small RNA profiling reveals phosphorus deficiency as a contributing factor in symptom expression for citrus huanglongbing disease.

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Zhao, Hongwei; Sun, Ruobai; Albrecht, Ute; Padmanabhan, Chellappan; Wang, Airong; Coffey, Michael D; Girke, Thomas; Wang, Zonghua; Close, Timothy J; Roose, Mikeal; Yokomi, Raymond K; Folimonova, Svetlana; Vidalakis, Georgios; Rouse, Robert; Bowman, Kim D; Jin, Hailing

2013-03-01

Huanglongbing (HLB) is a devastating citrus disease that is associated with bacteria of the genus 'Candidatus Liberibacter' (Ca. L.). Powerful diagnostic tools and management strategies are desired to control HLB. Host small RNAs (sRNA) play a vital role in regulating host responses to pathogen infection and are used as early diagnostic markers for many human diseases, including cancers. To determine whether citrus sRNAs regulate host responses to HLB, sRNAs were profiled from Citrus sinensis 10 and 14 weeks post grafting with Ca. L. asiaticus (Las)-positive or healthy tissue. Ten new microRNAs (miRNAs), 76 conserved miRNAs, and many small interfering RNAs (siRNAs) were discovered. Several miRNAs and siRNAs were highly induced by Las infection, and can be potentially developed into early diagnosis markers of HLB. miR399, which is induced by phosphorus starvation in other plant species, was induced specifically by infection of Las but not Spiroplasma citri that causes citrus stubborn-a disease with symptoms similar to HLB. We found a 35% reduction of phosphorus in Las-positive citrus trees compared to healthy trees. Applying phosphorus oxyanion solutions to HLB-positive sweet orange trees reduced HLB symptom severity and significantly improved fruit production during a 3-year field trial in south-west Florida. Our molecular, physiological, and field data suggest that phosphorus deficiency is linked to HLB disease symptomology.

Genetic similarity among Agistemus pallinii Matioli et al (Acari: Stigmaeidae) found in citrus orchards in Vicosa, Minas Gerais state, Brazil

Energy Technology Data Exchange (ETDEWEB)

Matioli, Andre L. [Instituto Biologico, Campinas, SP (Brazil). Lab. de Entomologia Economica], e-mail: almatioli@biologico.sp.gov.br; Pallini, Angelo [Universidade Federal de Vicosa (UFV), MG (Brazil). Dept. de Biologia Animal], e-mail: pallini@ufv.br; Tavares, Mara G. [Universidade Federal de Vicosa (UFV), MG (Brazil). Dept. de Biologia Geral], e-mail: mtavares@ufv.br

2009-03-15

Stigmaeidae are very important predators of mite and insect pests on several crops in Brazil. It is considered the second most important family of predatory mites in citrus orchards in Brazil. However, their identification, especially that of the members of the genus Agistemus, is rather difficult based only on morphology. Hence, this study describes the use of random amplified polymorphic DNA (RAPD-PCR) markers to determine the genetic similarity of an Agistemus pallinii Matioli et al population found in 2004 in a citrus orchard in Vicosa, Minas Gerais State, Brazil, preying on Panonychus citri (McGregor). Amplifications were performed with 12 random primers (OPAA8, OPAA19, OPAB1, OPAB5, OPAB18, OPAC9, OPAC17, OPAC19, OPAD10, OPAE9, OPAE12 and OPAE17), which generated 119 bands, with 53.8% polymorphism. The coefficients of genetic similarity among the individuals ranged from 0.68 to 0.99, indicating a high genetic similarity among them. The 3D projection analysis clustered the majority of individuals confirming their high similarity. Though individuals of A. pallinii are minute ({+-} 360 {mu}m long), the PCR-RAPD technique can still be used for their identification, complementing morphological analyses or for comparison of populations collected in different geographic regions. This is the fi rst molecular study carried out with stigmaeid mites. (author)

Antimicrobial constituents of the leaves of Mikania micrantha H. B. K.

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Yan Li

Full Text Available BACKGROUND: To isolate plant-derived compounds with antimicrobial activity from the leaves of Mikania micrantha, to determine the compounds configuration, and to evaluate their antimicrobial activity against eight plant pathogenic fungi (Exserohilum turcicum, Colletotrichum lagenarium, Pseudoperonispora cubensis, Botrytis cirerea, Rhizoctonia solani, Phytophthora parasitica, Fusarium solani, and Pythium aphanidermatum, and four plant pathogenic bacteria (gram negative bacteria: Ralstonia dolaanacearum, Xanthomonas oryzae pv. Oryzae, Xanthomonas Campestris pv. Vesicatoria, and Xanthomonas campestris pv. Citri, and four bacteria (gram positive bacteria: Staphyloccocus aureus, Bacillus subtilis, Micrococcus luteus, and Bacillus cereus. METHODS AND RESULTS: Antimicrobial constituents of the leaves of M. micrantha were isolated using bioactivity- guided fractionation. The antifungal activity of the isolated compounds was evaluated by the inhibit hypha growth method and inhibit spore germination method. Characterization of antibacterial activity was carried out using the minimum inhibitory concentrations (MICs and the minimum bactericidal concentrations (MBCs. MIC and MBC were determined by the broth microdilution method. Six compounds - deoxymikanolide, scandenolide, dihydroscandenolide, mikanolide, dihydromikanolide, and m - methoxy benzoic acid - have been isolated from leaves of Mikania micrantha H. B. K. Deoxymikanolide, scandenolide, and dihydroscandenolide were new compounds. The result of bioassay showed that all of isolated compounds were effective against tested strains and deoxymikanolide showed the strongest activity. CONCLUSIONS AND SIGNIFICANCE: The leaves of M. micrantha may be a promising source in the search for new antimicrobial drugs due to its efficacy and the broadest range. Meanwhile, adverse impact of M. micrantha will be eliminated.

Molecular characterization of three heat shock protein 70 genes and their expression profiles under thermal stress in the citrus red mite.

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Yang, Li-Hong; Jiang, Hong-Bo; Liu, Yong-Hua; Dou, Wei; Wang, Jin-Jun

2012-04-01

Three heat shock protein 70 family transcripts, named PcHsp70-1, PcHsp70-2 and PcHsp70-3, were isolated from the citrus red mite, Panonychus citri. PcHsp70-1, PcHsp70-2, and PcHsp70-3 contained an open reading frame of 1977, 1968, and 2028 nucleotides that encoded 658, 655 and 675 amino acid residues, respectively. Comparison of deduced amino acid sequences of PcHsp70-1 and PcHsp70-2 showed 86.34% identity, while the amino acid sequence of PcHsp70-3 was only 57.39 and 58.75% identical to that of PcHsp70-1 and PcHsp70-2, respectively. Sequences and phylogenetic analyses suggested that PcHsp70-1 and PcHsp70-2 were cytosolic Hsps, whereas PcHsp70-3 was located in ER (endoplasmic reticulum). To accurately validate mRNA expression profiles of the three Hsp70s under thermal stress conditions, seven housekeeping genes were evaluated. Alpha-tubulin and RpII were selected as optimal endogenous references for cold shock and heat shock conditions, respectively. Real-time quantitative RT-PCR revealed that only the mRNA expression of PcHsp70-2 was up-regulated under heat shocks, and all of the three Hsp70s were constitutively expressed under cold shocks. The results suggest that the three Hsp70s were more critical to coping with heat than cold shocks.

Sex allocation in a species with paternal genome elimination : The roles of crowding and female age in the mealybug Planococcus citri

NARCIS (Netherlands)

Ross, Laura; Langenhof, Minke B. W.; Pen, Ido; Beukeboom, Leo W.; West, Stuart A.; Shuker, David M.

Background: In species with paternal genome elimination, both sexes are diploid. However, in males the chromosomes inherited from the father are deactivated during early development and eliminated from the germ line. Sex allocation theory predicts that, all else being equal, females should bias

1H HRMAS NMR spectroscopy and chemometrics for evaluation of metabolic changes in citrus sinensis Caused by Xanthomonas axonopodis pv. citri

International Nuclear Information System (INIS)

Silva, Lorena M.A.; Alves Filho, Elenilson G.; Choze, Rafael; Liao, Luciano M.; Alcantara, Glaucia B.

2012-01-01

Xanthomonas axonopodis (Xac) bacterium causes one of the most feared and untreatable diseases in citriculture: citrus canker. To understand the response mechanisms of orange trees when attacked by Xac, leaves and fruits of Citrus sinensis were directly evaluated by HRMAS NMR (high resolution magic angle spinning nuclear magnetic resonance) spectroscopy. This technique allows the analysis of samples without laborious pre-treatments and also allows access to important information about chemical composition of samples. The orange tree leaves and fruit peels investigated in this study demonstrated the biochemical changes caused by Xac. Aided by chemometric analysis, the HRMAS NMR results show relevant changes in amino acids, carbohydrates, organic acids and terpenoids content. (author)

Disentangling mite predator-prey relationships by multiplex PCR.

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Pérez-Sayas, Consuelo; Pina, Tatiana; Gómez-Martínez, María A; Camañes, Gemma; Ibáñez-Gual, María V; Jaques, Josep A; Hurtado, Mónica A

2015-11-01

Gut content analysis using molecular techniques can help elucidate predator-prey relationships in situations in which other methodologies are not feasible, such as in the case of trophic interactions between minute species such as mites. We designed species-specific primers for a mite community occurring in Spanish citrus orchards comprising two herbivores, the Tetranychidae Tetranychus urticae and Panonychus citri, and six predatory mites belonging to the Phytoseiidae family; these predatory mites are considered to be these herbivores' main biological control agents. These primers were successfully multiplexed in a single PCR to test the range of predators feeding on each of the two prey species. We estimated prey DNA detectability success over time (DS50), which depended on the predator-prey combination and ranged from 0.2 to 18 h. These values were further used to weight prey detection in field samples to disentangle the predatory role played by the most abundant predators (i.e. Euseius stipulatus and Phytoseiulus persimilis). The corrected predation value for E. stipulatus was significantly higher than for P. persimilis. However, because this 1.5-fold difference was less than that observed regarding their sevenfold difference in abundance, we conclude that P. persimilis is the most effective predator in the system; it preyed on tetranychids almost five times more frequently than E. stipulatus did. The present results demonstrate that molecular tools are appropriate to unravel predator-prey interactions in tiny species such as mites, which include important agricultural pests and their predators. © 2015 John Wiley & Sons Ltd.

Effects of Euseius stipulatus on establishment and efficacy in spider mite suppression of Neoseiulus californicus and Phytoseiulus persimilis in clementine.

Science.gov (United States)

Abad-Moyano, Raquel; Urbaneja, Alberto; Hoffmann, Daniela; Schausberger, Peter

2010-04-01

The two-spotted spider mite, Tetranychus urticae, is one of the most problematic phytophagous pests in Spanish clementine orchards. The most abundant predatory mites in this ecosystem are Euseius stipulatus, Phytoseiulus persimilis and Neoseiulus californicus. Euseius stipulatus is dominant but poorly adapted to utilize T. urticae as prey. It mainly persists on pollen and citrus red mite, Panonychus citri. A recent study suggested that the more efficacious T. urticae predators P. persimilis and N. californicus are negatively affected by lethal and non-lethal intraguild interactions with E. stipulatus. Here, we investigated the potential of N. californicus and P. persimilis to colonize and thrive on young clementine trees infested by T. urticae in presence and absence of E. stipulatus. Presence of E. stipulatus interfered with establishment and abundance of P. persimilis and negatively affected the efficacy of N. californicus in T. urticae suppression. In contrast, the abundance of E. stipulatus was not affected by introduction of a second predator. Trait-mediated effects of E. stipulatus changing P. persimilis and N. californicus behavior and/or life history were the most likely explanations for these outcomes. We conclude that superiority of E. stipulatus in intraguild interactions may indeed contribute to the currently observed predator species composition and abundance, rendering natural control of T. urticae in Spanish clementine orchards unsatisfactory. Nonetheless, stronger reduction of T. urticae and/or plant damage in the predator combination treatments as compared to E. stipulatus alone indicates the possibility to improve T. urticae control via repeated releases of N. californicus and/or P. persimilis.

Multilocus Sequence Typing Reveals Relevant Genetic Variation and Different Evolutionary Dynamics among Strains of Xanthomonas arboricola pv. juglandis

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Marco Scortichini

2010-11-01

Full Text Available Forty-five Xanthomonas arboricola pv. juglandis (Xaj strains originating from Juglans regia cultivation in different countries were molecularly typed by means of MultiLocus Sequence Typing (MLST, using acnB, gapA, gyrB and rpoD gene fragments. A total of 2.5 kilobases was used to infer the phylogenetic relationship among the strains and possible recombination events. Haplotype diversity, linkage disequilibrium analysis, selection tests, gene flow estimates and codon adaptation index were also assessed. The dendrograms built by maximum likelihood with concatenated nucleotide and amino acid sequences revealed two major and two minor phylotypes. The same haplotype was found in strains originating from different continents, and different haplotypes were found in strains isolated in the same year from the same location. A recombination breakpoint was detected within the rpoD gene fragment. At the pathovar level, the Xaj populations studied here are clonal and under neutral selection. However, four Xaj strains isolated from walnut fruits with apical necrosis are under diversifying selection, suggesting a possible new adaptation. Gene flow estimates do not support the hypothesis of geographic isolation of the strains, even though the genetic diversity between the strains increases as the geographic distance between them increases. A triplet deletion, causing the absence of valine, was found in the rpoD fragment of all 45 Xaj strains when compared with X. axonopodis pv. citri strain 306. The codon adaptation index was high in all four genes studied, indicating a relevant metabolic activity.

Profiling and analysis of multiple constituents in Baizhu Shaoyao San before and after processing by stir-frying using UHPLC/Q-TOF-MS/MS coupled with multivariate statistical analysis.

Science.gov (United States)

Xu, Yangyang; Cai, Hao; Cao, Gang; Duan, Yu; Pei, Ke; Tu, Sicong; Zhou, Jia; Xie, Li; Sun, Dongdong; Zhao, Jiayu; Liu, Jing; Wang, Xiaoqi; Shen, Lin

2018-04-15

Baizhu Shaoyao San (BSS) is a famous traditional Chinese medicinal formula widely used for the treatment of painful diarrhea, intestinal inflammation, and diarrhea-predominant irritable bowel syndrome. According to clinical medication, three medicinal herbs (Atractylodis Macrocephalae Rhizoma, Paeoniae Radix Alba, and Citri Reticulatae Pericarpium) included in BSS must be processed using some specific methods of stir-frying. On the basis of the classical theories of traditional Chinese medicine, the therapeutic effects of BSS would be significantly enhanced after processing. Generally, the changes of curative effects mainly result from the variations of inside chemical basis caused by the processing procedure. To find out the corresponding changes of chemical compositions in BSS after processing and to elucidate the material basis of the changed curative effects, an optimized ultra-high-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry in positive and negative ion modes coupled with multivariate statistical analyses were developed. As a result, a total of 186 compounds were ultimately identified in crude and processed BSS, in which 62 marker compounds with significant differences between crude and processed BSS were found by principal component analysis and t-test. Compared with crude BSS, the contents of 23 compounds were remarkably decreased and the contents of 39 compounds showed notable increase in processed BSS. The transformation mechanisms of some changed compounds were appropriately inferred from the results. Furthermore, compounds with extremely significant differences might strengthen the effects of the whole herbal formula. Copyright © 2018 Elsevier B.V. All rights reserved.

Genomics-enabled analysis of the emergent disease cotton bacterial blight.

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Anne Z Phillips

2017-09-01

Full Text Available Cotton bacterial blight (CBB, an important disease of (Gossypium hirsutum in the early 20th century, had been controlled by resistant germplasm for over half a century. Recently, CBB re-emerged as an agronomic problem in the United States. Here, we report analysis of cotton variety planting statistics that indicate a steady increase in the percentage of susceptible cotton varieties grown each year since 2009. Phylogenetic analysis revealed that strains from the current outbreak cluster with race 18 Xanthomonas citri pv. malvacearum (Xcm strains. Illumina based draft genomes were generated for thirteen Xcm isolates and analyzed along with 4 previously published Xcm genomes. These genomes encode 24 conserved and nine variable type three effectors. Strains in the race 18 clade contain 3 to 5 more effectors than other Xcm strains. SMRT sequencing of two geographically and temporally diverse strains of Xcm yielded circular chromosomes and accompanying plasmids. These genomes encode eight and thirteen distinct transcription activator-like effector genes. RNA-sequencing revealed 52 genes induced within two cotton cultivars by both tested Xcm strains. This gene list includes a homeologous pair of genes, with homology to the known susceptibility gene, MLO. In contrast, the two strains of Xcm induce different clade III SWEET sugar transporters. Subsequent genome wide analysis revealed patterns in the overall expression of homeologous gene pairs in cotton after inoculation by Xcm. These data reveal important insights into the Xcm-G. hirsutum disease complex and strategies for future development of resistant cultivars.

Genomes-based phylogeny of the genus Xanthomonas

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Rodriguez-R Luis M

2012-03-01

Full Text Available Abstract Background The genus Xanthomonas comprises several plant pathogenic bacteria affecting a wide range of hosts. Despite the economic, industrial and biological importance of Xanthomonas, the classification and phylogenetic relationships within the genus are still under active debate. Some of the relationships between pathovars and species have not been thoroughly clarified, with old pathovars becoming new species. A change in the genus name has been recently suggested for Xanthomonas albilineans, an early branching species currently located in this genus, but a thorough phylogenomic reconstruction would aid in solving these and other discrepancies in this genus. Results Here we report the results of the genome-wide analysis of DNA sequences from 989 orthologous groups from 17 Xanthomonas spp. genomes available to date, representing all major lineages within the genus. The phylogenetic and computational analyses used in this study have been automated in a Perl package designated Unus, which provides a framework for phylogenomic analyses which can be applied to other datasets at the genomic level. Unus can also be easily incorporated into other phylogenomic pipelines. Conclusions Our phylogeny agrees with previous phylogenetic topologies on the genus, but revealed that the genomes of Xanthomonas citri and Xanthomonas fuscans belong to the same species, and that of Xanthomonas albilineans is basal to the joint clade of Xanthomonas and Xylella fastidiosa. Genome reduction was identified in the species Xanthomonas vasicola in addition to the previously identified reduction in Xanthomonas albilineans. Lateral gene transfer was also observed in two gene clusters.

Study of the distribution patterns of the constituent herbs in classical Chinese medicine prescriptions treating respiratory disease by data mining methods.

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Fu, Xian-Jun; Song, Xu-Xia; Wei, Lin-Bo; Wang, Zhen-Guo

2013-08-01

To provide the distribution pattern and compatibility laws of the constituent herbs in prescriptions, for doctor's convenience to make decision in choosing correct herbs and prescriptions for treating respiratory disease. Classical prescriptions treating respiratory disease were selected from authoritative prescription books. Data mining methods (frequent itemsets and association rules) were used to analyze the regular patterns and compatibility laws of the constituent herbs in the selected prescriptions. A total of 562 prescriptions were selected to be studied. The result exhibited that, Radix glycyrrhizae was the most frequently used in 47.2% prescriptions, other frequently used were Semen armeniacae amarum, Fructus schisandrae Chinese, Herba ephedrae, and Radix ginseng. Herbal ephedrae was always coupled with Semen armeniacae amarum with the confidence of 73.3%, and many herbs were always accompanied by Radix glycyrrhizae with high confidence. More over, Fructus schisandrae Chinese, Herba ephedrae and Rhizoma pinelliae was most commonly used to treat cough, dyspnoea and associated sputum respectively besides Radix glycyrrhizae and Semen armeniacae amarum. The prescriptions treating dyspnoea often used double herb group of Herba ephedrae & Radix glycyrrhizae, while prescriptions treating sputum often used double herb group of Rhizoma pinelliae & Radix glycyrrhizae and Rhizoma pinelliae & Semen armeniacae amarum, triple herb groups of Rhizoma pinelliae & Semen armeniacae amarum & Radix glycyrrhizae and Pericarpium citri reticulatae & Rhizoma pinelliae & Radix glycyrrhizae. The prescriptions treating respiratory disease showed common compatibility laws in using herbs and special compatibility laws for treating different respiratory symptoms. These principle patterns and special compatibility laws reported here could be useful for doctors to choose correct herbs and prescriptions in treating respiratory disease.

Effects of Radiation on Germ Cells of Insects: Dominant Lethals, Gamete Inactivation and Gonial-Cell Killing; Effets des rayonnements sur les cellules germinales des insectes: letalite dominante, inactivation des gametes et destruction des cellules des gonades; Vozdejstvie radiatsii na polovye kletki nasekomykh: dominantnye letali, inaktivatsiya gamet i umershchvlenie polovykh kletok; Efectos de las radiaciones sobre las celulas germinales de los insectos: elementos letales dominantes, inactivacion de los gametos y exterminacion de las celulas gonadicas

Energy Technology Data Exchange (ETDEWEB)

Von Borstel, R. C. [Biology Division, Oak Ridge National Laboratory, Oak Ridge, TN (United States)

1963-09-15

Radiations and chemical mutagens kill cells in numerous ways: by one of several kinds of induced dominant lethality, by a direct inactivation of function as with sperm, and by genetically undefinable types of death which may or may not be related to dominant lethality per se. Also, chemical mutagens appear to exert a curious enhancement of the fertilizing capacity of sperm. The different stages of oflgenesis and spermatogenesis respond with unequal sensitivity to radiation, and individual cells pass through stages conferring as much as a 50-fold difference in sensitivity. Where species of Diptera, Hymenoptera and Coleoptera can be compared, a striking similarity of response to radiation can be observed, both to stage sensitivity and degree of response with dose. The silkworm, Bombyx mori (Lepidoptera), seems to be similar in most respects to representatives of the other orders in response of germ cells to radiation, but differs sharply in types of dominant lethality induced. Species having atypical genetic mechanisms (e.g.,the lecanoid system of Planococcus citri (Hemipt. : Coccidae) are special cases, and their responses to radiation are considerably modified from those of other species. For insect population control by the irradiation-of-male method, dominant lethality is as advantageous in species where matings are multiple as in species where mating occurs once. Sperm inactivation and gonial killing can be regarded as instances of true sterility and are maximally effective only in species where mating occurs once. For most efficient control, doses should be chosen which would induce maximum dominant lethality, minimum sperm inactivation and complete killing of gonial cells. These parameters are simple to determine by gamete viability measurements, irradiated and unirradiated population competition experiments and histological examination of gonia. (author) [French] Les rayonnements et les agents chimiques de mutation detruisent les cellules de nombreuses facons

Mycoplasmas, plants, insect vectors: a matrimonial triangle.

Science.gov (United States)

Garnier, M; Foissac, X; Gaurivaud, P; Laigret, F; Renaudin, J; Saillard, C; Bové, J M

2001-10-01

Plant pathogenic mycoplasmas were discovered by electron microscopy, in 1967, long after the discovery and culture in 1898 of the first pathogenic mycoplasma of animal origin, Mycoplasma mycoides. Mycoplasmas are Eubacteria of the class Mollicutes, a group of organisms phylogenetically related to Gram-positive bacteria. Their more characteristic features reside in the small size of their genomes, the low guanine (G) plus cytosine (C) content of their genomic DNA and the lack of a cell wall. Plant pathogenic mycoplasmas are responsible for several hundred diseases and belong to two groups: the phytoplasmas and the spiroplasmas. The phytoplasmas (previously called MLOs, for mycoplasma like organisms) were discovered first; they are pleiomorphic, and have so far resisted in vitro cultivation. Phytoplasmas represent the largest group of plant pathogenic Mollicutes. Only three plant pathogenic spiroplasmas are known today. Spiroplasma citri, the agent of citrus stubborn was discovered and cultured in 1970 and shown to be helical and motile. S. kunkelii is the causal agent of corn stunt. S. phoeniceum, responsible for periwinkle yellows, was discovered in Syria. There are many other spiroplasmas associated with insects and ticks. Plant pathogenic mycoplasmas are restricted to the phloem sieve tubes in which circulates the photosynthetically-enriched sap, the food for many phloem-feeding insects (aphids, leafhoppers, psyllids, etc.). Interestingly, phytopathogenic mycoplasmas are very specifically transmitted by leafhoppers or psyllid species. In this paper, the most recent knowledge on phytopathogenic mycoplasmas in relation with their insect and plant habitats is presented as well as the experiments carried out to control plant mycoplasma diseases, by expression of mycoplasma-directed-antibodies in plants (plantibodies).

Bioefficacy of a novel calcium-potassium salt of (-)-hydroxycitric acid

International Nuclear Information System (INIS)

Downs, Bernard W.; Bagchi, Manashi; Subbaraju, Gottumukkala V.; Shara, Michael A.; Preuss, Harry G.; Bagchi, Debasis

2005-01-01

Obesity is associated with cardiovascular disease, diabetes and certain forms of cancer. Popular strategies on weight loss often fail to address many key factors such as fat mass, muscle density, bone density, water mass, their inter-relationships and impact on energy production, body composition, and overall health and well-being. (-)-Hydroxycitric acid (HCA), a natural plant extract from the dried fruit rind of Garcinia cambogia, has been reported to promote body fat loss in humans without stimulating the central nervous system. The level of effectiveness of G. cambogia extract is typically attributed solely to HCA. However, other components by their presence or absence may significantly contribute to its therapeutic effectiveness. Typically, HCA used in dietary weight loss supplement is bound to calcium, which results in a poorly soluble ( 2+ /K + bound (-)-HCA salt (HCA-SX or Super CitriMax) make it completely water soluble as well as bioavailable. An efficacious dosage of HCA-SX (4500 mg/day t.i.d.) provides a good source of Ca 2+ (495 mg, 49.5% of RDI) and K + (720 mg, 15% of RDI). Ca 2+ ions are involved in weight management by increasing lipid metabolism, enhancing thermogenesis, and increasing bone density. K + , on the other hand, increases energy, reduces hypertension, increases muscle strength and regulates arrhythmias. Both Ca and K act as buffers in pH homeostasis. HCA-SX has been shown to increase serotonin availability, reduce appetite, increase fat oxidation, improve blood lipid levels, reduce body weight, and modulate a number of obesity regulatory genes without affecting the mitochondrial and nuclear proteins required for normal biochemical and physiological functions

Ectopic expression of MdSPDS1 in sweet orange (Citrus sinensis Osbeck) reduces canker susceptibility: involvement of H₂O₂ production and transcriptional alteration.

Science.gov (United States)

Fu, Xing-Zheng; Chen, Chuan-Wu; Wang, Yin; Liu, Ji-Hong; Moriguchi, Takaya

2011-03-28

Enormous work has shown that polyamines are involved in a variety of physiological processes, but information is scarce on the potential of modifying disease response through genetic transformation of a polyamine biosynthetic gene. In the present work, an apple spermidine synthase gene (MdSPDS1) was introduced into sweet orange (Citrus sinensis Osbeck 'Anliucheng') via Agrobacterium-mediated transformation of embryogenic calluses. Two transgenic lines (TG4 and TG9) varied in the transgene expression and cellular endogenous polyamine contents. Pinprick inoculation demonstrated that the transgenic lines were less susceptible to Xanthomonas axonopodis pv. citri (Xac), the causal agent of citrus canker, than the wild type plants (WT). In addition, our data showed that upon Xac attack TG9 had significantly higher free spermine (Spm) and polyamine oxidase (PAO) activity when compared with the WT, concurrent with an apparent hypersensitive response and the accumulation of more H₂O₂. Pretreatment of TG9 leaves with guazatine acetate, an inhibitor of PAO, repressed PAO activity and reduced H₂O₂ accumulation, leading to more conspicuous disease symptoms than the controls when both were challenged with Xac. Moreover, mRNA levels of most of the defense-related genes involved in synthesis of pathogenesis-related protein and jasmonic acid were upregulated in TG9 than in the WT regardless of Xac infection. Our results demonstrated that overexpression of the MdSPDS1 gene prominently lowered the sensitivity of the transgenic plants to canker. This may be, at least partially, correlated with the generation of more H₂O₂ due to increased production of polyamines and enhanced PAO-mediated catabolism, triggering hypersensitive response or activation of defense-related genes.

Priming by Hexanoic acid induce activation of mevalonic and linolenic pathways and promotes the emission of plant volatiles.

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Eugenio eLlorens

2016-04-01

Full Text Available Hexanoic acid is a short natural monocarboxylic acid present in some fruits and plants. Previous studies reported that soil drench application of this acid induces effective resistance in tomato plants against Botrytis cinerea and Pseudomonas syringae and in citrus against Alternaria alternata and Xanthomonas citri. In this work, we performed an in deep study of the metabolic changes produced in citrus by the application of hexanoic acid in response to the challenge pathogen Alternaria alternata, focusing on the response of the plant. Moreover, we used 13C labeled hexanoic to analyze its behavior inside the plants. Finally, we studied the volatile emission of the treated plants after the challenge inoculation. Drench application of 13C labeled hexanoic demonstrated that this molecule stays in the roots and is not mobilized to the leaves, suggesting long distance induction of resistance. Moreover, the study of the metabolic profile showed an alteration of more than two hundred molecules differentially induced by the application of the compound and the inoculation with the fungus. Bioinformatics analysis of data showed that most of these altered molecules could be related with the mevalonic and linolenic pathways suggesting the implication of these pathways in the induced resistance mediated by hexanoic acid. Finally, the application of this compound showed an enhancement of the emission of 17 volatile metabolites. Taken together, this study indicates that after the application of hexanoic acid this compound remains in the roots, provoking molecular changes that may trigger the defensive response in the rest of the plant mediated by changes in the mevalonic and linolenic pathways and enhancing the emission of volatile compounds, suggesting for the first time the implication of mevalonic pathway in response to hexanoic application.

The TAL effector PthA4 interacts with nuclear factors involved in RNA-dependent processes including a HMG protein that selectively binds poly(U RNA.

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Tiago Antonio de Souza

Full Text Available Plant pathogenic bacteria utilize an array of effector proteins to cause disease. Among them, transcriptional activator-like (TAL effectors are unusual in the sense that they modulate transcription in the host. Although target genes and DNA specificity of TAL effectors have been elucidated, how TAL proteins control host transcription is poorly understood. Previously, we showed that the Xanthomonas citri TAL effectors, PthAs 2 and 3, preferentially targeted a citrus protein complex associated with transcription control and DNA repair. To extend our knowledge on the mode of action of PthAs, we have identified new protein targets of the PthA4 variant, required to elicit canker on citrus. Here we show that all the PthA4-interacting proteins are DNA and/or RNA-binding factors implicated in chromatin remodeling and repair, gene regulation and mRNA stabilization/modification. The majority of these proteins, including a structural maintenance of chromosomes protein (CsSMC, a translin-associated factor X (CsTRAX, a VirE2-interacting protein (CsVIP2, a high mobility group (CsHMG and two poly(A-binding proteins (CsPABP1 and 2, interacted with each other, suggesting that they assemble into a multiprotein complex. CsHMG was shown to bind DNA and to interact with the invariable leucine-rich repeat region of PthAs. Surprisingly, both CsHMG and PthA4 interacted with PABP1 and 2 and showed selective binding to poly(U RNA, a property that is novel among HMGs and TAL effectors. Given that homologs of CsHMG, CsPABP1, CsPABP2, CsSMC and CsTRAX in other organisms assemble into protein complexes to regulate mRNA stability and translation, we suggest a novel role of TAL effectors in mRNA processing and translational control.

Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1.

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Zhang, Shujian; Chakrabarty, Pranjib K; Fleites, Laura A; Rayside, Patricia A; Hopkins, Donald L; Gabriel, Dean W

2015-01-01

Xylella fastidiosa (X. fastidiosa) infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD) of grapevines. X. fastidiosa biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703) and a serine protease (PD0956); two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928), and at least one relatively short, hemagglutinin-like protein (PD0986). Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of E. coli BL21(DE3) overexpressing PD1703 exhibited a hypersensitive response (HR) in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to Xanthomonas citri, E. coli and X. fastidiosa EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot) and a PD0986 (hemagglutinin) were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on V. vinifera vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of X. fastidiosa populations in grapevines or elderberry.

Effect of insecticides on mealybug destroyer (Coleoptera: Coccinellidae) and parasitoid Leptomastix dactylopii (Hymenoptera: Encyrtidae), natural enemies of citrus mealybug (Homoptera: Pseudococcidae).

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Cloyd, Raymond A; Dickinson, Amy

2006-10-01

In this study, we measured, under laboratory conditions, the direct and indirect effects of insecticides on mealybug destroyer, Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae), and parasitoid Leptomastix dactylopii Howard (Hymenoptera: Encyrtidae), natural enemies of citrus mealybug, Planococcus citri (Risso) (Homoptera: Pseudococcidae). The adult stages of both natural enemies were exposed to sprays of the insecticides buprofezin, pyriproxyfen, flonicamid, acetamiprid, dinotefuran, and clothianidin at label-recommended rates to assess direct mortality after 24, 48, and 72 h, respectively. The effects of the insecticides on L. dactylopii parasitization rate and percentage of parasitoid emergence also were monitored using the label and 4x the recommended label rate. Dinotefuran was extremely detrimental to the adult parasitoid at the label rate with 100% mortality after 24 h. Buprofezin, pyriproxyfen, and flonicamid were not harmful to L. dactylopii when applied at the label rate. At 4x the recommended label rate, dinotefuran, acetamiprid, and clothianidin were all harmful to the parasitoid with 100% mortality 72 h after application. Both buprofezin and flonicamid were not toxic to L. dactylopii with 100% adult survival after 72 h. Pyriproxyfen and flonicamid, at both the label and 4x the recommended label rate, did not negatively affect L. dactylopii parasitization rate or percentage of parasitoid emergence. Acetamiprid, dinotefuran, and clothianidin were toxic to C. montrouzieri adults with 100% mortality after 48 h, whereas buprofezin, pyriproxyfen, and flonicamid demonstrated minimal (10-20% mortality after 48 h) harmful effects to the predator. Based on the results from our study, the indirect effects of the insect growth regulator (IGR) buprofezin were not decisive; however, the IGR pyriproxyfen and the insecticide flonicamid were not directly or indirectly harmful to the predator C. montrouzieri and parastioid L. dactylopii, indicating that

Bioefficacy of a novel calcium-potassium salt of (-)-hydroxycitric acid

Energy Technology Data Exchange (ETDEWEB)

Downs, Bernard W. [InterHealth Research Center, Benicia, CA (United States); Bagchi, Manashi [InterHealth Research Center, Benicia, CA (United States); Subbaraju, Gottumukkala V. [Laila Impex Research Center, Vijayawada (India); Shara, Michael A. [Department of Pharmacy Sciences, Creighton University Medical Center, 2500 California Plaza, Omaha, 68178 NE (United States); Preuss, Harry G. [Department of Physiology and Biophysics, Georgetown University Medical Center, Washington, DC (United States); Bagchi, Debasis [InterHealth Research Center, Benicia, CA (United States) and Department of Pharmacy Sciences, Creighton University Medical Center, 2500 California Plaza, Omaha, 68178 NE (United States)]. E-mail: debsis@creighton.edu

2005-11-11

Obesity is associated with cardiovascular disease, diabetes and certain forms of cancer. Popular strategies on weight loss often fail to address many key factors such as fat mass, muscle density, bone density, water mass, their inter-relationships and impact on energy production, body composition, and overall health and well-being. (-)-Hydroxycitric acid (HCA), a natural plant extract from the dried fruit rind of Garcinia cambogia, has been reported to promote body fat loss in humans without stimulating the central nervous system. The level of effectiveness of G. cambogia extract is typically attributed solely to HCA. However, other components by their presence or absence may significantly contribute to its therapeutic effectiveness. Typically, HCA used in dietary weight loss supplement is bound to calcium, which results in a poorly soluble (<50%) and less bioavailable form. Conversely, the structural characteristics of a novel Ca{sup 2+}/K{sup +} bound (-)-HCA salt (HCA-SX or Super CitriMax) make it completely water soluble as well as bioavailable. An efficacious dosage of HCA-SX (4500 mg/day t.i.d.) provides a good source of Ca{sup 2+} (495 mg, 49.5% of RDI) and K{sup +} (720 mg, 15% of RDI). Ca{sup 2+} ions are involved in weight management by increasing lipid metabolism, enhancing thermogenesis, and increasing bone density. K{sup +}, on the other hand, increases energy, reduces hypertension, increases muscle strength and regulates arrhythmias. Both Ca and K act as buffers in pH homeostasis. HCA-SX has been shown to increase serotonin availability, reduce appetite, increase fat oxidation, improve blood lipid levels, reduce body weight, and modulate a number of obesity regulatory genes without affecting the mitochondrial and nuclear proteins required for normal biochemical and physiological functions.

Ectopic expression of MdSPDS1 in sweet orange (Citrus sinensis Osbeck reduces canker susceptibility: involvement of H2O2 production and transcriptional alteration

Directory of Open Access Journals (Sweden)

Wang Yin

2011-03-01

Full Text Available Abstract Background Enormous work has shown that polyamines are involved in a variety of physiological processes, but information is scarce on the potential of modifying disease response through genetic transformation of a polyamine biosynthetic gene. Results In the present work, an apple spermidine synthase gene (MdSPDS1 was introduced into sweet orange (Citrus sinensis Osbeck 'Anliucheng' via Agrobacterium-mediated transformation of embryogenic calluses. Two transgenic lines (TG4 and TG9 varied in the transgene expression and cellular endogenous polyamine contents. Pinprick inoculation demonstrated that the transgenic lines were less susceptible to Xanthomonas axonopodis pv. citri (Xac, the causal agent of citrus canker, than the wild type plants (WT. In addition, our data showed that upon Xac attack TG9 had significantly higher free spermine (Spm and polyamine oxidase (PAO activity when compared with the WT, concurrent with an apparent hypersensitive response and the accumulation of more H2O2. Pretreatment of TG9 leaves with guazatine acetate, an inhibitor of PAO, repressed PAO activity and reduced H2O2 accumulation, leading to more conspicuous disease symptoms than the controls when both were challenged with Xac. Moreover, mRNA levels of most of the defense-related genes involved in synthesis of pathogenesis-related protein and jasmonic acid were upregulated in TG9 than in the WT regardless of Xac infection. Conclusion Our results demonstrated that overexpression of the MdSPDS1 gene prominently lowered the sensitivity of the transgenic plants to canker. This may be, at least partially, correlated with the generation of more H2O2 due to increased production of polyamines and enhanced PAO-mediated catabolism, triggering hypersensitive response or activation of defense-related genes.

Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1.

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Shujian Zhang

Full Text Available Xylella fastidiosa (X. fastidiosa infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD of grapevines. X. fastidiosa biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703 and a serine protease (PD0956; two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928, and at least one relatively short, hemagglutinin-like protein (PD0986. Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of E. coli BL21(DE3 overexpressing PD1703 exhibited a hypersensitive response (HR in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to Xanthomonas citri, E. coli and X. fastidiosa EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot and a PD0986 (hemagglutinin were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on V. vinifera vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of X. fastidiosa populations in grapevines or elderberry.

Cercosporoid fungi (Mycosphaerellaceae) 2. Species on monocots (Acoraceae to Xyridaceae, excluding Poaceae).

Science.gov (United States)

Braun, Uwe; Crous, Pedro W; Nakashima, Chiharu

2014-12-01

Cercosporoid fungi (formerly Cercospora s. lat.) represent one of the largest groups of hyphomycetes belonging to the Mycosphaerellaceae (Ascomycota). They include asexual morphs, asexual holomorphs, or species with mycosphaerella-like sexual morphs. Most of them are leaf-spotting plant pathogens with special phytopathological relevance. In the first part of a new monographic work, cercosporoid hyphomycetes occurring on other fungi (fungicolous species), on ferns (pteridophytes) and gymnosperms were treated. This second part deals with cercosporoid fungi on monocots (Liliopsida; Equisetopsida, Magnoliidae, Lilianae), which covers species occurring on host plants belonging to families arranged in alphabetical order from Acoraceae to Xyridaceae, excluding Poaceae (cereals and grasses) which requires a separate treatment. The species are described and illustrated in alphabetical order under the particular cercosporoid genera, supplemented by keys to the species concerned. A detailed introduction, a survey of currently recognised cercosporoid genera, a key to the genera concerned, and a discussion of taxonomically relevant characters were published in the first part of this series. Neopseudocercospora, an additional recently introduced cercosporoid genus, is briefly discussed. The following taxonomic novelties are introduced: Cercospora alpiniigena sp. nov., C. neomaricae sp. nov., Corynespora palmicola comb. nov., Exosporium miyakei comb. nov., E. petersii comb. nov., Neopseudocercospora zambiensis comb. nov., Passalora caladiicola comb. nov., P. streptopi comb. nov., P. togashiana comb. nov., P. tranzschelii var. chinensis var. nov., Pseudocercospora beaucarneae comb. nov., P. constrictoflexuosa comb. et stat. nov., P. curcumicola sp. nov., P. dispori comb. nov., P. smilacicola sp. nov., P. urariigena nom. nov., Zasmidium agavicola comb. nov., Z. cercestidis-afzelii comb. nov., Z. citri-griseum comb. nov., Z. cyrtopodii comb. nov., Z. gahnae comb. nov., Z. indicum

The signal peptide-like segment of hpaXm is required for its association to the cell wall in transgenic tobacco plants.

Science.gov (United States)

Li, Le; Miao, Weiguo; Liu, Wenbo; Zhang, Shujian

2017-01-01

Harpins, encoded by hrp (hypersensitive response and pathogenicity) genes of Gram-negative plant pathogens, are elicitors of hypersensitive response (HR). HpaXm is a novel harpin-like protein described from cotton leaf blight bacteria, Xanthomonas citri subsp. malvacearum-a synonym of X. campestris pv. malvacearum (Smith 1901-1978). A putative signal peptide (1-MNSLNTQIGANSSFL-15) of hpaXm was predicted in the nitroxyl-terminal (N-terminal)by SignalP (SignalP 3.0 server). Here, we explored the function of the N-terminal leader peptide like segment of hpaXm using transgenic tobacco (Nicotiana tabacum cv. Xanthi nc.). Transgenic tobacco lines expressing the full-length hpaXm and the signal peptide-like segment-deleted mutant hpaXmΔLP were developed using transformation mediated by Agrobacterium tumefaciens. The target genes were confirmed integrated into the tobacco genomes and expressed normally. Using immune colloidal-gold detection technique, hpaXm protein was found to be transferred to the cytoplasm, the cell membrane, and organelles such as chloroplasts, mitochondria, and nucleus, as well as the cell wall. However, the deletion mutant hpaXmΔLP expressed in transgenic tobacco was found unable to cross the membrane to reach the cell wall. Additionally, soluble proteins extracted from plants transformed with hpaXm and hpaXmΔLP were bio-active. Defensive micro-HR induced by the transgene expression of hpaXm and hpaXmΔLP were observed on transgenic tobacco leaves. Disease resistance bioassays to tobacco mosaic virus (TMV) showed that tobacco plants transformed with hpaXm and with hpaXmΔLP exhibited enhanced resistance to TMV. In summary, the N-terminal signal peptide-like segment (1-45 bp) in hpaXm sequence is not necessary for transgene expression, bioactivity of hpaXm and resistance to TMV in transgenic tobacco, but is required for the protein to be translocated to the cell wall.

Isolation and identification of citrus psorosis virus Egyptian isolate (CPsV-EG).

Science.gov (United States)

Ghazal, S A; El-Dougdoug, Kh A; Mousa, A A; Fahmy, H; Sofy, A R

2008-01-01

Citrus psorosis ophiovirus (CPsV), is considered to be of the most serious and deter mental virus pathogen's citrus species trees in Egypt. CPsV-EG was isolated from infected citrus grapefruit (C. paradisi Macf.) at Agric. Res. Centre (ARC). The grapefruit which used for CPsV-EG isolate was found to be free from CTV, CEVd and Spiroplasma citri where as gave -ve results with DTBIA, tissue print hybridization and Diene's stain respectively. CPsV-EG was detected on the basis of biological indexing by graft inoculation which gave oak leaf pattern (OLP) on Dweet tangor and serological assay by DAS-ELISA using Mab specific CPsV. CPsV-EG was reacted with variable responses on 16 host plants belonging to 6 families. Only 8 host plants are susceptible and showed visible external symptoms which appeared as local, systemic and local followed by systemic infections. CPsV-EG isolate was transmitted from infected citrus to citrus by syringe and grafting and herbaceous plants by forefinger inoculation and syringe. The woody indicators and rootstocks were differed in response to CPsV-EG isolate which appeared as no-response, response, sensitivity and hypersensitivity. The serological characters represented as the antigenic determinants of CPsV-EG isolate related to monoclonal antibodies specific CPsV strain where as appeared precipitation reaction by DAS-ELISA and DTBIA. The partial fragment of RNA3 (coat protein gene) of CPsV-EG (-1140bp and -571bp) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from grapefruit tissues using two sets primers specific CPsV (CPV3 and CPV4) and (PS66 and PS65) respectively. The virus under study was identified as CPsV-EG isolate according to biological, serological and molecular characters.

The role of horizontal transfer in the evolution of a highly variable lipopolysaccharide biosynthesis locus in xanthomonads that infect rice, citrus and crucifers

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Bogdanove Adam J

2007-12-01

Full Text Available Abstract Background Lipopolysaccharide (LPS is a pathogen associated molecular pattern (PAMP of animal and plant pathogenic bacteria. Variation at the interstrain level is common in LPS biosynthetic gene clusters of animal pathogenic bacteria. This variation has been proposed to play a role in evading the host immune system. Even though LPS is a modulator of plant defense responses, reports of interstrain variation in LPS gene clusters of plant pathogenic bacteria are rare. Results In this study we report the complete sequence of a variant 19.9 kb LPS locus present in the BXO8 strain of Xanthomonas oryzae pv. oryzae (Xoo, the bacterial blight pathogen of rice. This region is completely different in size, number and organization of genes from the LPS locus present in most other strains of Xoo from India and Asia. Surprisingly, except for one ORF, all the other ORFs at the BXO8 LPS locus are orthologous to the genes present at this locus in a sequenced strain of X. axonopodis pv. citri (Xac; a pathogen of citrus plants. One end of the BXO8 LPS gene cluster, comprised of ten genes, is also present in the related rice pathogen, X. oryzae pv. oryzicola (Xoc. In Xoc, the remainder of the LPS gene cluster, consisting of seven genes, is novel and unrelated to LPS gene clusters of any of the sequenced xanthomonads. We also report substantial interstrain variation suggestive of very recent horizontal gene transfer (HGT at the LPS biosynthetic locus of Xanthomonas campestris pv. campestris (Xcc, the black rot pathogen of crucifers. Conclusion Our analyses indicate that HGT has altered the LPS locus during the evolution of Xanthomonas oryzae pathovars and suggest that the ancestor of all Xanthomonas oryzae pathovars had an Xac type of LPS gene cluster. Our finding of interstrain variation in two major xanthomonad pathogens infecting different hosts suggests that the LPS locus in plant pathogenic bacteria, as in animal pathogens, is under intense

Transformation of sweet orange [Citrus sinensis (L.) Osbeck] with pthA-nls for acquiring resistance to citrus canker disease.

Science.gov (United States)

Yang, Li; Hu, Chunhua; Li, Na; Zhang, Jiayin; Yan, Jiawen; Deng, Ziniu

2011-01-01

The COOH terminal of pthA encoding three nuclear localizing signals (NLS) was amplified by polymerase chain reaction (PCR) from the plasmid of Xanthomonas axonopodis pv. citri, the pathogen of citrus canker disease. Then the sense and antisense strands of the nls were cloned into pBI121 vector. pthA-nls driven by the CaMV35 s promoter was transferred into sweet orange via Agrobacterium -mediated transformation. Successful integration was confirmed by PCR and Southern blotting, and 12 sense-nls (nls (+)) and 9 antisense-nls (nls (-)) transgenic clones were obtained. The expression of nls fragment was analyzed by RT-PCR, Real time q-PCR and Western blotting, in which the specific NLS protein was detected only in nls (+) transgenic clones. In an in vitro assay, when pin-puncture inoculation was performed with 2.5 × 10(7) cfu/ml of bacterial solution, the nls (+) transgenic clones showed no typical lesion development, while typical symptoms were observed in the wild types and the nls (-) transgenic clones. In vivo assay results indicated that the nls (+) transgenic clones showed less disease incidence, in comparison with the wild types and the nls (-) transgenic clones, when pin-puncture inoculation was performed with 10(4)-10(5) cfu/ml. The minimum disease incidence was 23.3% for 'Sucarri' sweet orange and 33.3% for 'Bingtang' sweet orange. When 10(4)-10(7) cfu/ml of pathogen was spray inoculated, the nls (+) transgenic clones did not show any symptom, and even the concentration raised to 10(9) cfu/ml, the disease incidence was 20-80%, while the wild types and the nls (-) transgenic clones had 100% disease development with whatever concentration of inoculum. Two transgenic clones were confirmed to be resistant to citrus canker disease in the repeated inoculation. The results suggested that the transformation of nls sense strands may offer an effective way to acquire resistance to citrus canker disease.

Investigation of a Quadruplex-Forming Repeat Sequence Highly Enriched in Xanthomonas and Nostoc sp.

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Charlotte Rehm

Full Text Available In prokaryotes simple sequence repeats (SSRs with unit sizes of 1-5 nucleotides (nt are causative for phase and antigenic variation. Although an increased abundance of heptameric repeats was noticed in bacteria, reports about SSRs of 6-9 nt are rare. In particular G-rich repeat sequences with the propensity to fold into G-quadruplex (G4 structures have received little attention. In silico analysis of prokaryotic genomes show putative G4 forming sequences to be abundant. This report focuses on a surprisingly enriched G-rich repeat of the type GGGNATC in Xanthomonas and cyanobacteria such as Nostoc. We studied in detail the genomes of Xanthomonas campestris pv. campestris ATCC 33913 (Xcc, Xanthomonas axonopodis pv. citri str. 306 (Xac, and Nostoc sp. strain PCC7120 (Ana. In all three organisms repeats are spread all over the genome with an over-representation in non-coding regions. Extensive variation of the number of repetitive units was observed with repeat numbers ranging from two up to 26 units. However a clear preference for four units was detected. The strong bias for four units coincides with the requirement of four consecutive G-tracts for G4 formation. Evidence for G4 formation of the consensus repeat sequences was found in biophysical studies utilizing CD spectroscopy. The G-rich repeats are preferably located between aligned open reading frames (ORFs and are under-represented in coding regions or between divergent ORFs. The G-rich repeats are preferentially located within a distance of 50 bp upstream of an ORF on the anti-sense strand or within 50 bp from the stop codon on the sense strand. Analysis of whole transcriptome sequence data showed that the majority of repeat sequences are transcribed. The genetic loci in the vicinity of repeat regions show increased genomic stability. In conclusion, we introduce and characterize a special class of highly abundant and wide-spread quadruplex-forming repeat sequences in bacteria.

Investigation of a Quadruplex-Forming Repeat Sequence Highly Enriched in Xanthomonas and Nostoc sp.

Science.gov (United States)

Rehm, Charlotte; Wurmthaler, Lena A; Li, Yuanhao; Frickey, Tancred; Hartig, Jörg S

2015-01-01

In prokaryotes simple sequence repeats (SSRs) with unit sizes of 1-5 nucleotides (nt) are causative for phase and antigenic variation. Although an increased abundance of heptameric repeats was noticed in bacteria, reports about SSRs of 6-9 nt are rare. In particular G-rich repeat sequences with the propensity to fold into G-quadruplex (G4) structures have received little attention. In silico analysis of prokaryotic genomes show putative G4 forming sequences to be abundant. This report focuses on a surprisingly enriched G-rich repeat of the type GGGNATC in Xanthomonas and cyanobacteria such as Nostoc. We studied in detail the genomes of Xanthomonas campestris pv. campestris ATCC 33913 (Xcc), Xanthomonas axonopodis pv. citri str. 306 (Xac), and Nostoc sp. strain PCC7120 (Ana). In all three organisms repeats are spread all over the genome with an over-representation in non-coding regions. Extensive variation of the number of repetitive units was observed with repeat numbers ranging from two up to 26 units. However a clear preference for four units was detected. The strong bias for four units coincides with the requirement of four consecutive G-tracts for G4 formation. Evidence for G4 formation of the consensus repeat sequences was found in biophysical studies utilizing CD spectroscopy. The G-rich repeats are preferably located between aligned open reading frames (ORFs) and are under-represented in coding regions or between divergent ORFs. The G-rich repeats are preferentially located within a distance of 50 bp upstream of an ORF on the anti-sense strand or within 50 bp from the stop codon on the sense strand. Analysis of whole transcriptome sequence data showed that the majority of repeat sequences are transcribed. The genetic loci in the vicinity of repeat regions show increased genomic stability. In conclusion, we introduce and characterize a special class of highly abundant and wide-spread quadruplex-forming repeat sequences in bacteria.

Bio-based synthesis of silver nanoparticles from orange waste: effects of distinct biomolecule coatings on size, morphology, and antimicrobial activity.

Science.gov (United States)

de Barros, Caio Henrique Nasi; Cruz, Guilherme Crispim Faria; Mayrink, Willian; Tasic, Ljubica

2018-01-01

Despite the numerous reports on biological syntheses of silver nanoparticles (AgNPs), little is known about the composition of their capping agents, protein corona of plant extract-mediated synthesis, and their influence on the properties of AgNPs. Here, orange ( Citrus sinensis ) waste was utilized as a source of an extract for AgNP synthesis (the protein corona composition of which was elucidated), and also as a starting material for hesperidin and nanocellulose extraction, which were used for bio-based AgNP synthesis. A comparison of the results using the two methods of synthesis is presented. AgNPs were synthesized using orange ( C. sinensis ) peel extract (Or-AgNPs) in a biological route, and using hesperidin (Hsd-AgNPs) and nanocellulose (extracted from oranges) in a green chemical route. Characterization of nanoparticles was carried out using zeta potential and hydrodynamic size measurements, transmission electron microscopy, and X-ray diffraction. Elucidation of proteins from protein corona was performed via ultra performance liquid chromatography-tandem mass spectrometer experiments. Antimicrobial activity was assessed via minimum inhibitory concentration assays against Xanthomonas axonopodis pv. citri ( Xac ), the bacterium that causes citric canker in oranges. Or-AgNPs were not completely uniform in morphology, having a size of 48.1±20.5 nm and a zeta potential of -19.0±0.4 mV. Stabilization was performed mainly by three proteins, which were identified by tandem mass spectrometry (MS/MS) experiments. Hsd-AgNPs were smaller (25.4±12.5 nm) and had uniform morphology. Nanocellulose provided a strong steric and electrostatic (-28.2±1.0 mV) stabilization to the nanoparticles. Both AgNPs presented roughly the same activity against Xac , with the minimum inhibitory concentration range between 22 and 24 μg mL -1 . Despite the fact that different capping biomolecules on AgNPs had an influence on morphology, size, and stability of AgNPs, the antibacterial