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Sample records for psyllids diaphorina citri

  1. Complete Genome Sequence of a Putative Densovirus of the Asian Citrus Psyllid, Diaphorina citri.

    Science.gov (United States)

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

    2016-07-28

    Here, we report the complete genome sequence of a putative densovirus of the Asian citrus psyllid, Diaphorina citri Diaphorina citri densovirus (DcDNV) was originally identified through metagenomics, and here, we obtained the complete nucleotide sequence using PCR-based approaches. Phylogenetic analysis places DcDNV between viruses of the Ambidensovirus and Iteradensovirus genera.

  2. Complete Genome Sequence of Diaphorina citri-associated C virus, a Novel Putative RNA Virus of the Asian Citrus Psyllid, Diaphorina citri.

    Science.gov (United States)

    Nouri, Shahideh; Salem, Nidà; Falk, Bryce W

    2016-07-21

    We present here the complete nucleotide sequence and genome organization of a novel putative RNA virus identified in field populations of the Asian citrus psyllid, Diaphorina citri, through sequencing of the transcriptome followed by reverse transcription-PCR (RT-PCR). We tentatively named this virus Diaphorina citri-associated C virus (DcACV). DcACV is an unclassified positive-sense RNA virus.

  3. Gender differences and effect of photophase on Asian citrus psyllid (Diaphorina citri Kuwayama) feeding behavior

    Science.gov (United States)

    Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), thought to be primarily a phloem-feeding insect, transmits the presumptive pathogen for Huanglongbing, ‘Candidatus Liberibacter asiaticus’. Because this bacterium is restricted to the phloem and bacterial transmission is the res...

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

    Science.gov (United States)

    Huanglongbing (HLB) is the most destructive disease of citrus worldwide. 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. Laboratory and field studies were cond...

  5. Correlation of an electrical penetration graph waveform with walking by Asian citrus psyllid, Diaphorina citri Kuwayama

    Science.gov (United States)

    Electrical penetration graph (EPG) monitoring is being used to study how feeding behavior of the Asian citrus psyllid (ACP) (Diaphorina citri Kuwayama) changes in response to insecticides, in an effort to improve the efficacy of widespread insecticide treatments against the insect in Florida citrus....

  6. Endosymbiont hunting in the metagenome of Asian citrus psyllid (Diaphorina citri) (7th Annual SFAF Meeting, 2012)

    Energy Technology Data Exchange (ETDEWEB)

    Saha, Surya [Cornell University

    2012-06-01

    Surya Saha on "Endosymbiont hunting in the metagenome of Asian citrus psyllid (Diaphorina citri)" at the 2012 Sequencing, Finishing, Analysis in the Future Meeting held June 5-7, 2012 in Santa Fe, New Mexico.

  7. Titers of 'Ca. Liberibacter asiaticus' in Murraya paniculata and Murraya-reared Diaphorina citri Are Much Lower than in Citrus and Citrus-reared Psyllids

    National Research Council Canada - National Science Library

    Walter, Abigail; Duan, Y; Hall, David G

    2012-01-01

    Huanglongbing, one of the most devastating diseases of citrus, is associated with the bacterium 'Candidatus Liberibacter asiaticus' vectored by the Asian citrus psyllid, Diaphorina citri, in North America...

  8. Repellent Activity of Botanical Oils against Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae).

    Science.gov (United States)

    Kuhns, Emily H; Martini, Xavier; Hoyte, Angel; Stelinski, Lukasz L

    2016-07-14

    The Asian citrus psyllid, Diaphorina citri Kuwayama, is the insect vector of the pathogen causing huanglongbing. We selected three botanical oils to evaluate behavioral activity against D. citri. In laboratory olfactometer assays, fir oil was repellent to D. citri females, while litsea and citronella oils elicited no response from D. citri females. In choice settling experiments, D. citri settled almost completely on control plants rather than on plants treated with fir oil at a 9.5 mg/day release rate. Therefore, we conducted field trials to determine if fir oil reduced D. citri densities in citrus groves. We found no repellency of D. citri from sweet orange resets that were treated with fir oil dispensers releasing 10.4 g/day/tree as compared with control plots. However, we found a two-week decrease in populations of D. citri as compared with controls when the deployment rate of these dispensers was doubled. Our results suggest that treatment of citrus with fir oil may have limited activity as a stand-alone management tool for D. citri and would require integration with other management practices.

  9. Repellent Activity of Botanical Oils against Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae

    Directory of Open Access Journals (Sweden)

    Emily H. Kuhns

    2016-07-01

    Full Text Available The Asian citrus psyllid, Diaphorina citri Kuwayama, is the insect vector of the pathogen causing huanglongbing. We selected three botanical oils to evaluate behavioral activity against D. citri. In laboratory olfactometer assays, fir oil was repellent to D. citri females, while litsea and citronella oils elicited no response from D. citri females. In choice settling experiments, D. citri settled almost completely on control plants rather than on plants treated with fir oil at a 9.5 mg/day release rate. Therefore, we conducted field trials to determine if fir oil reduced D. citri densities in citrus groves. We found no repellency of D. citri from sweet orange resets that were treated with fir oil dispensers releasing 10.4 g/day/tree as compared with control plots. However, we found a two-week decrease in populations of D. citri as compared with controls when the deployment rate of these dispensers was doubled. Our results suggest that treatment of citrus with fir oil may have limited activity as a stand-alone management tool for D. citri and would require integration with other management practices.

  10. Geographic Distribution of Habitat, Development, and Population Growth Rates of the Asian Citrus Psyllid, Diaphorina citri, in Mexico

    OpenAIRE

    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...

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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. PMID:28278188

  14. Toxicity of insecticidal soaps to the Asian citrus psyllid (Diaphorina citri) and two of its natural enemies

    Science.gov (United States)

    Insecticidal soaps (hereafter “soaps”) are labeled for use by homeowners and also can be used in citrus grown for the organic market. Soaps control some insect pests and therefore might be an alternative to conventional pesticides for control of the Asian citrus psyllid, Diaphorina citri Kuwayama (...

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

    Institute of Scientific and Technical Information of China (English)

    Raiinder S.Mann; Russell L.Rouseff; Jack Smoot; Nandikeswara Rao; Wendy L.Meyer; Stephen L.Lapointe; Paul S.Robbins

    2013-01-01

    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.cirri 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).

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

    Directory of Open Access Journals (Sweden)

    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.

  17. Molecular characterization of Wolbachia strains associated with the invasive Asian citrus psyllid Diaphorina citri in Brazil.

    Science.gov (United States)

    Guidolin, A S; Cônsoli, F L

    2013-02-01

    Wolbachia is a symbiont intensively studied due to its ability to interfere with their host's reproduction, and it has been recently proposed as an alternative tool to control insect pests or vectors of diseases. The Asian citrus psyllid Diaphorina citri is an important pest of citrus since it vectors the bacterium that causes the "Huanglongbing" disease in citrus. The frequency and diversity of Wolbachia associated with D. citri is unknown, limiting the utilization of Wolbachia as an alternative strategy for insect management. Thus, we aimed to determine the natural rate of infection, to characterize the Wolbachia strains associated with this psyllid by "multilocus sequencing typing" (MLST) and wsp analysis, and to verify the association of the symbiont to particular genotypes of the host. Analysis indicated Wolbachia infects 100 % of all specimens tested from all 15 sampled populations. MLST revealed the occurrence of five new sequence types (STs) of Wolbachia, while analysis based on the wsp sequences indicated only four different types of Wolbachia. ST-173 was predominant, while the remaining STs were population specific. Analysis of the host-symbiont relationship did not reveal any particular association of Wolbachia and haplotypes or a decrease in nucleotide diversity of D. citri in populations in which more than one ST was recorded. The consequences of the diversity of STs reported are still unknown, but the fact that Wolbachia infection is fixed and that there is one ST with a broad distribution highlights the use of this symbiont as an alternative strategy to control D. citri.

  18. Multimodal cues drive host-plant assessment in Asian citrus psyllid (Diaphorina citri).

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    Patt, Joseph M; Meikle, William G; Mafra-Neto, Agenor; Sétamou, Mamoudou; Mangan, Robert; Yang, Chenghai; Malik, Nasir; Adamczyk, John J

    2011-12-01

    Asian citrus psyllid (Diaphorina citri) transmits the causal agent of Huanglongbing, a devastating disease of citrus trees. In this study we measured behavioral responses of D. citri to combinations of visual, olfactory, and gustatory stimuli in test arenas. Stimuli were presented to the psyllids in droplets or lines of an emulsified wax formulation in two different arena types in no-choice tests. First, when placed on a colored ring situated halfway between the center and perimeter of a petri dish, D. citri spent more time on yellow versus gray rings; however, this response disappeared when either gray or yellow wax droplets were applied. When the psyllids were presented with droplets scented with terpenes, the response to both scent and color was increased. The addition of a dilute (≍0.1 M) sucrose solution to the wax droplets increased the magnitude of D. citri responses. Next, groups of D. citri were placed on plastic laboratory film covering a sucrose solution, to mimic a leaf surface. Test stimuli were presented via two 'midribs' made from lines of emulsified wax formulation. Probing levels were measured as a function of color saturation and scent composition, and concentration. The test scents were based on qualitatively major volatiles emitted by Murraya paniculata (L.) Jack, Citrus aurantifolia (Christm.) Swingle, and C. sinensis (L.) Osbeck. The highest probing response was observed on the middle concentration (20-μl scent/10 ml wax formulation) of the C. aurantifolia-scented wax lines. Results indicate that there are interactive effects between the different sensory modalities in directing host-plant assessment behavior.

  19. 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

    Science.gov (United States)

    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...

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

    Directory of Open Access Journals (Sweden)

    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.

  1. Seasonal Population Dynamics of the Asian Citrus Psyllid, Diaphorina citri Kuwayama in Sarawak

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    Stephen L.C. Teck

    2011-01-01

    Full Text Available Problem statement: Effective control of phytophagous pests requires a thorough understanding of their seasonal population dynamics, dispersion behavior, natural enemy activity and climate. To date, although very little detail information had been published on the ecology of Diaphorina citri. The objective of this investigation was to test through field experiment the hypothesis that the major factors influencing local D. citri populations particularly their seasonal population dynamics in Sarawak are (a flushing cycles, (b climate and (c the impact of the primary parasitoids namely Tamarixia radiata and Diaphorencyrtus aligarhensis. Approach: Seasonal abundance D. citri was studied weekly from March 1998 to December 2000 in the 1-ha citrus honey mandarin (Citrus aurantium L. commercial orchard at Jemukan (1° 33'N, 110° 41'E, Kota Samarahan Division, Southwest Sarawak, in Malaysia. Results: Field studies on citrus trees showed that the D. citri population fluctuates throughout the year on citrus honey mandarin in Sarawak. Generations overlapped but adult and egg population peaks for a short period generally coincided with three annual flushing cycles, in August-September, February-March and June-July between March 1998 and December 2000. Conclusion: Psyllid population levels are positively related to the availability of new shoot flushes. Psyllid populations are adversely affected by weather conditions and parasitoids. Adult psyllid populations increased exponentially during periods of flush growth and migration and dispersal of the adults was also related to flushing cycles. Dispersal and colonization of new trees is greatest in September-October, at the onset of the rainy season.

  2. 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.

  3. Insecticide sprays, natural enemy assemblages and predation on Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae).

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    Monzo, C; Qureshi, J A; Stansly, P A

    2014-10-01

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama is considered a key citrus pest due to its role as vector of 'huanglongbing' (HLB) or citrus greening, probably the most economically damaging disease of citrus. Insecticidal control of the vector is still considered a cornerstone of HLB management to prevent infection and to reduce reinoculation of infected trees. The severity of HLB has driven implementation of intensive insecticide programs against ACP with unknown side effects on beneficial arthropod fauna in citrus agroecosystems. We evaluated effects of calendar sprays directed against this pest on natural enemy assemblages and used exclusion to estimate mortality they imposed on ACP populations in citrus groves. Predator exclusion techniques were used on nascent colonies of D. citri in replicated large untreated and sprayed plots of citrus during the four major flushing periods over 2 years. Population of spiders, arboreal ants and ladybeetles were independently assessed. Monthly sprays of recommended insecticides for control of ACP, adversely affected natural enemy populations resulting in reduced predation on ACP immature stages, especially during the critical late winter/early spring flush. Consequently, projected growth rates of the ACP population were greatest where natural enemies had been adversely affected by insecticides. Whereas, this result does not obviate the need for insecticidal control of ACP, it does indicate that even a selective regimen of sprays can impose as yet undetermined costs in terms of reduced biological control of this and probably other citrus pests.

  4. Complete Genome Sequence of the Largest Known Flavi-Like Virus, Diaphorina citri flavi-like virus, a Novel Virus of the Asian Citrus Psyllid, Diaphorina citri.

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    Matsumura, Emilyn E; Nerva, Luca; Nigg, Jared C; Falk, Bryce W; Nouri, Shahideh

    2016-09-08

    A novel flavi-like virus tentatively named Diaphorina citri flavi-like virus (DcFLV) was identified in field populations of Diaphorina citri through small RNA and transcriptome sequencing followed by reverse transcription (RT)-PCR. We report here the complete nucleotide sequence and genome organization of DcFLV, the largest flavi-like virus identified to date.

  5. Complete Genome Sequence of the Largest Known Flavi-Like Virus, Diaphorina citri flavi-like virus, a Novel Virus of the Asian Citrus Psyllid, Diaphorina citri

    Science.gov (United States)

    Matsumura, Emilyn E.; Nerva, Luca; Nigg, Jared C.; Falk, Bryce W.

    2016-01-01

    A novel flavi-like virus tentatively named Diaphorina citri flavi-like virus (DcFLV) was identified in field populations of Diaphorina citri through small RNA and transcriptome sequencing followed by reverse transcription (RT)-PCR. We report here the complete nucleotide sequence and genome organization of DcFLV, the largest flavi-like virus identified to date. PMID:27609921

  6. 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

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

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    Iliano V Coutinho-Abreu

    Full Text Available BACKGROUND: 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. RESULTS: 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%. CONCLUSION: 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.

  8. Influence of Host Plant Species and Flush Growth Stage on the Asian Citrus Psyllid, Diaphorina citri Kuwayama

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    Stephen L.C. Teck

    2011-01-01

    Full Text Available Problem statement: In order to establish the important host range of D. citri, the performance of this psyllid on different rutacous host plants was assessed. Apparent regional differences in host preferences of D. citri as noted above and the need for empirical knowledge of its host preferences in Malaysia where it is lacking. Approach: Experiments on host plant preference between citrus, jasmine orange and curry leaf were conducted over a period of 14 days in the field cage in October 1998 at the agricultural research centre, Semonggok, 15 km from Kuching, Kuching Division, Sarawak in Malaysia. Results: A comparative life cycle study of the Asian Citrus Psyllid Diaphorina citri Kuwayama on three different Rutaceous plant hosts namely citrus, jasmine orange and curry leaf plants was conducted inside the temperature controlled growth chamber maintain at 26 ± 1°C temperature, 75-80% relative humidity with artificial illumination intensity of 10,000 lux equipped with a photoperiod of 14:10 h light: dark regime to permit normal growth. Conclusion: The screenhouse studies confirmed that D. citri can colonise and breed on citrus, jasmine orange and curry leaf plant and that jasmine orange is the preferred host. It is also confirmed that females prefer to oviposit on immature flush 3-10 mm in length. In growth chamber experiments the life cycle of the psyllid was 18.5 days on jasmine orange, 19.0 days on citrus and 23.0 days on curry leaf.

  9. Titers of 'Ca. Liberibacter asiaticus' in Murraya paniculata and Murraya-reared Diaphorina citri are much lower than in citrus and citrus-reared psyllids

    Science.gov (United States)

    Huanglongbing, one of the most devastating diseases of citrus, is associated with the bacterium Candidatus Liberibacter asiaticus vectored by the Asian citrus psyllid, Diaphorina citri, in North America. Murraya paniculata is a common ornamental plant that is an alternate host of Ca. L. asiaticus an...

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

    Science.gov (United States)

    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...

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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

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

    Directory of Open Access Journals (Sweden)

    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

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    Galdeano, Diogo Manzano; Breton, Michèle Claire; Lopes, João Roberto Spotti; Falk, Bryce W.

    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. PMID:28282380

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

    Directory of Open Access Journals (Sweden)

    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.

  17. Nested-quantitative PCR approach with improved sensitivity for the detection of low titer levels of Candidatus Liberibacter asiaticus in the Asian citrus psyllid, Diaphorina citri Kuwayama.

    Science.gov (United States)

    Coy, M R; Hoffmann, M; Kingdom Gibbard, H N; Kuhns, E H; Pelz-Stelinski, K S; Stelinski, L L

    2014-07-01

    Candidatus Liberibacter asiaticus (CLas) is a phloem-limited bacterium transmitted by the Asian citrus psyllid, Diaphorina citri, and the presumptive causal agent of citrus greening disease. The current method of detection for CLas within plant and insect samples is by a presence/absence qPCR assay using the CLas 16S rDNA gene target. Although qPCR is highly sensitive, low bacterial titers or suboptimal qPCR conditions can result in false-negatives. Using a nested qPCR assay, we determined the false-negative rate of the 16S presence/absence qPCR assay was greater than 50%. Studies to determine the performance parameters of the qPCR assays for CLas 16S and Wingless (Wg), the D. citri endogenous gene, using plasmid and psyllid DNA, revealed suboptimal and variable performance of the 16S assay in psyllid samples. Average efficiencies and sensitivity limits of the plasmid assays were 99.0% and 2.7 copies of template for Wg, respectively, and 98.5% and 2.2-22.1 copies for 16S, respectively. Variability in efficiency was significantly greater in psyllid samples for both gene targets compared to the corresponding plasmid assays, and efficiencies as low as 76% were obtained for 16S. A secondary structure analysis revealed the formation of two stem-loop structures that block the forward and probe binding sites in the 16S template, which could hinder amplification. In summary, our results suggest that suboptimal qPCR efficiency is not uncommon for the 16S presence/absence qPCR assay, which combined with lowCLas titers in some samples, could contribute significantly to the under-reporting of CLas infection in psyllid and plant samples.

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

    Science.gov (United States)

    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...

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

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

    2017-01-23

    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, LD50 values were determined and ranged from 0.10 to 0.53 ng/μl for imidacloprid, fenpropathrin, dimethoate and spinetoram. LD50 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  3. The complete mitochondrial genome sequence of Diaphorina citri (Hemiptera: Psyllidae)

    Science.gov (United States)

    The first complete mitochondrial genome (mitogenome) sequence of Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae), from Guangzhou, China is presented. The circular mitogenome is 14,996 bp in length with an A+T content of 74.5%, and contains 13 protein-coding genes (PCGs), 22 tRNA genes ...

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

    Science.gov (United States)

    Diaphorina citri, the Asian citrus psyllid,(Hemiptera) is the insect vector of Candidatus Liberibacter asiaticus, the presumed cause of citrus greening disease, known as huanglongbing. Sequencing of the Diaphorina citri metagenome has been initiated to gain better understanding of the biology of D. ...

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

    Science.gov (United States)

    Ammar, El-Desouky; Hall, David G; Shatters, Robert G

    2013-01-01

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  7. Advances in Biological Control of Citrus Psyllid Diaphorina citri, a Vector Insect of Citrus Huanglongbing Disease%黄龙病媒介昆虫柑橘木虱生物防治新进展

    Institute of Scientific and Technical Information of China (English)

    代晓彦; 任素丽; 周雅婷; 任顺祥; 邱宝利

    2014-01-01

    The citrus psyllid, Diaphorina citri Kuwayama, is the primary vector of citrus Huanglongbing disease, Candidatus Liberibacter asiaticus, which has the huge potential threats to the development of citrus industry in China. Based on the comprehensive understanding of D. citri control technologies, we reviewed the latest advances in biological control of D. citri and forecasted its future prospects. The main biological control resources of citrus psyllid currently are natural enemy insects and entomopathogenic fungi, among which predatory ladybeetles, lacewings, spiders and hover flies are the dominant species of predator, while Tamarixia radiata and Diaphorencyrtus aligarhensis are the two dominant parasitoids in China. Several representative entomopathogenic fungi include Lecanicillium lecanii, Isaria fumosorosea, Beauveria bassiana and Hirsutella citriformis have been reported with high virulence to citrus psyllid. This current review is expected to provide technical support and theoretical reference for efficient and sustainable control of citrus psyllid in the future.%柑橘木虱Diaphorina citri Kuwayama是柑橘黄龙病的重要媒介昆虫,对我国柑橘产业的健康发展潜在着巨大的威胁。本文在综合国内柑橘木虱防控技术的基础上,对近年来柑橘木虱生物防治的最新研究进展进行了综述,并且对其生物防治的发展前景给予了展望。目前应用于柑橘木虱生物防治的天敌资源主要是天敌昆虫及昆虫病原真菌,其中捕食性天敌主要包括瓢虫、食蚜蝇、草蛉和蜘蛛,寄生性天敌昆虫主要包括啮小蜂和跳小峰;昆虫病原真菌主要包括蜡蚧轮枝菌、拟青霉、白僵菌、桔形被毛孢等。期望通过对柑橘木虱生物防治工作的综述,为我国今后柑橘木虱的科学、高效防控及其可持续治理提供参考,不断推动我国柑橘产业的健康发展。

  8. 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.

  9. Sequencing and annotation of the Wolbachia endosymbiont of Diaphorina citri by the CG-HLB genome resources group reveals candidate sources of interaction with the insect host

    Science.gov (United States)

    Diaphorina citri (Hemiptera: Psyllidae), the Asian citrus psyllid, is the vector of Ca. Liberibacter asiaticus, the causal agent of huanglongbing. The D. citri metagenome has been completed to gain a better understanding of the biology of this organism and the potential roles of other psyllid bacter...

  10. Within-orchard edge effects of the azimuth of the sun on Diaphorina citri adults in mature orchards

    Science.gov (United States)

    Huanglongbing (HLB) has been considered the most devastating disease of citrus. The bacterium and vector associated with HLB in Florida are ‘Candidatus Liberibacter asiaticus’ and Diaphorina citri (Asian citrus psyllid), respectively. D. citri is positively phototropic, and higher populations have b...

  11. An improved method of DNA extraction from Diaphorina citri for HLB detection

    Science.gov (United States)

    Huanglongbing (HLB) is a devastating disease of citrus that is transmitted by two citrus psyllids. Diaphorina citri transmits Candidatus Liberibacter asiaticus (Las) and Ca. L. americanus (Lam), and Trioza erytreae transmits Ca. L. africanus (Laf). Ca. Liberibacter species can be detected in DNA ex...

  12. Population ecology and phenology of Diaphorina citri (Hemiptera: Psyllidae) in two Florida citrus groves

    Science.gov (United States)

    Studies were conducted during 2005 and 2006 to assess population densities and phenology of the psyllid Diaphorina citri Kuwayama in citrus within east central Florida. Young, irrigated grapefruit and mature, non-irrigated orange trees were sampled weekly for eggs, nymphs and adults on flush shoots;...

  13. Effects of temperature, photoperiod, and rainfall on morphometric variation of Diaphorina citri (Hemiptera: Liviidae)

    Science.gov (United States)

    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 artifi...

  14. Low incidence of Candidatus Liberibacter asiaticus in Diaphorina citri and its host plant Murraya paniculata

    Science.gov (United States)

    Huanglongbing (HLB) is one of the most devastating diseases of citrus worldwide. Candidatus Liberibacter asiaticus is the prevalent species of three HLB-associated Liberibacter species, which is vectored by the psyllid Diaphorina citri. The vector and the bacteria have host plants outside Citrus, ...

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

    Science.gov (United States)

    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.

  16. First record of Diaphorina citri (Hemiptera: Psyllidae) in Ecuador infesting urban citrus and orange jasmine trees.

    Science.gov (United States)

    Cornejo, J F; Chica, E J

    2014-01-01

    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.

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

    Science.gov (United States)

    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...

  18. Brachygastra mellifica (Hymenoptera: Vespidae): Predation preference and feeding behavior on Diaphorina citri (Hemiptera: Psyllidae) in Mexico

    OpenAIRE

    Reyes-Rosas, M. A.; Loera-Gallardo, J.; López-Arroyo, J. I.; Buck, M.

    2014-01-01

    In previous field studies in Northern Mexico, we found the wasp Brachygastra mellifica (Say 1837) (Hymenoptera: Vespidae) preying voraciously the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae), the vector of the bacteria Ca. Liberibacter spp., the putative agent of Huanglongbing, one of the most devastating citrus disease in the world.   As in Mexico, the ACP management considers the use of pest biological control, the availability of potential agents for the con...

  19. Presencia de Diaphorina citri (Hemiptera: Psyllidae en el Noroeste Argentino (NOA Record of Diaphorina citri (Hemiptera: Psyllidae in North West Argentina

    Directory of Open Access Journals (Sweden)

    Lucrecia Augier

    2006-12-01

    Full Text Available Se amplía la distribución de Diaphorina citri Kuwayama (Hemiptera: Psyllidae para el Noroeste Argentino.The distribution of Diaphorina citri Kuwayama (Hemiptera: Psyllidae is expanded to North West Argentina.

  20. Temporal response and attraction of Diaphorina citri to visual stimuli

    Science.gov (United States)

    As the vector of the global disease of citrus greening or huanglongbing, Asian citrus pysllids, Diaphorina citri Kuwayama (Hemiptera; Liviidae) are the greatest threat to the worldwide citrus industry. Critical to management of D. citri and huanglongbing, is optimization of surveillance methodologie...

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    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...

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

    Science.gov (United States)

    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...

  4. Entomophagous insects associated to Diaphorina citri (Hemiptera: Psyllidae) in citrus orchards with different weed management systems in Papantla, Veracruz, Mexico

    OpenAIRE

    Ortega-Arenas, L. D.; López-López, R.; Lomelí-Flores, J. R.; Cedillo-Portugal, E.; Gómez-Tovar, L.; Salazar-Cruz, J.; Villegas-Monter, A.

    2014-01-01

    Huanglongbing (HLB), one of the most destructive diseases of citrus worldwide, is threatening the survival of the citrus industry in Mexico. Diaphorina citri is the primary vector of HLB; thus, control of the vector it’s vital for disease management. This study was carried out to evaluate the influence of different management systems on the population psyllid density and entomophagous insects associated in orange orchards (Citrus sinensis cv. Valencia) in Papantla, Veracruz, Mexico. Five orch...

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  7. Biology of Diaphorina citri (Homoptera: Psyllidae) on four host plants.

    Science.gov (United States)

    Tsai, J H; Liu, Y H

    2000-12-01

    The biology of the citrus psyllid Diaphorina citri Kuwayama was studied at 25 degrees C on four commonly grown citrus and related plants [rough lemon, Citrus jambhiri Lush; sour orange, C aurantium L.; grapefruit, C. paradisi Macfadyen; and orange jessamine, Murraya paniculata (L.) Jack] in the laboratory. The biological characteristics of each life stage are described. The average egg incubation periods on orange jessamine, grapefruit, rough lemon, and sour orange varied very little (4.1-4.2 d). The average nymphal developmental periods on these four host plants were essentially the same except the fifth stadium. Survival of immatures on orange jessamine, grapefruit, rough lemon, and sour orange was 75.4, 84.6, 78.3, and 68.6%, respectively. Female adults lived an average of 39.7, 39.7, 47.6, and 43.7 d on these respective host plants. The average number of eggs laid per female on grapefruit (858 eggs) was significantly more than those on other hosts (P < 0.05). The intrinsic rate of natural increase (r(m)) for D. citri on grapefruit was highest. Jackknife estimates of r(m) varied from 0.188 on grapefruit to 0.162 on orange jessamine and rough lemon. The mean population generation time on these hosts ranged from 31.6 to 34.1 d. The continuous flushes produced by orange jessamine could play an important role in maintaining high populations of this vector when the new flushes are not available in the commercial citrus groves.

  8. Vibrational communication between the sexes in Diaphorina citri (Hemiptera: Psyllidae)

    Science.gov (United States)

    We examined the substrate-borne vibrational signals used in communication between the sexes in Diaphorina citri Kuwayama (Hemiptera: Psyllidae), a vector of huanglongbing (an economically devastating disease of citrus), in an anechoic chamber and an olfactometer. Males and females both primarily pro...

  9. 柑橘木虱两地理种群的内共生菌群落组成及传菌能力%The endosymbiotic microbiota of two geographical populations of the Asian citrus psyllid,Diaphorina citri,in China and its ability of pathogen transmission

    Institute of Scientific and Technical Information of China (English)

    马晓芳; 张学潮; 徐海君

    2012-01-01

    柑橘黄龙病(Huanglongbing,HLB)是经柑橘木虱Diaphorina citri传播的最主要柑橘病害之一,危害严重时能对柑橘产业造成毁灭性的破坏.为了鉴定福建和海南2个地理种群柑橘木虱的内共生菌群落组成,本研究对16SrRNA部分保守序列进行PCR扩增,并利用特异性引物对不同内共生菌进行了感染率检测;另外,还通过人工接虫的方法,探索柑橘木虱成虫在带黄龙病菌蕉柑Citrus reticulata cv.Tankan 上的获菌能力,以及带菌柑橘木虱成虫对黄岩蜜橘C.reticulata cv.Subcompress的传菌能力.研究发现,这2个地理种群的柑橘木虱含有相同的内共生菌组成,包括α-Proteobacteria,Wolbachia spp.,γ-Proteobacteria,mycetocyte symbionts,β-Proteobacteria,Oxalobacter 和β-Proteobacteria,Herbaspirillum,而且这2个地理种群柑橘木虱的4种内共生菌的携带率均在95%以上.柑橘木虱成虫在带菌蕉柑上饲菌28 d后,带菌率可达到82%,而带菌柑橘木虱成虫在黄岩蜜橘上传菌75 d后,可导致橘树整体带菌.本研究为柑橘木虱的进一步研究和防虫治病途径提供了一些理论依据.%Huanglongbing (HLB) is one of the most severe citrus diseases, which is usually transmitted by the Asian citrus psyllid ( Diaphorina citri), and causes great economic loss. To understand the endosyrnbiotic microbiota of the citrus psyllid from Hainan and Fujian provinces in China, the partial conserved region of 16S rRNA was amplified by PCR, and subsequently the infection rate of each endosymbiont from two geographical populations of the citrus psyllid was detected by PCR using specific primer pairs in this stduy. Moreover, we detected the acquisition efficiency of the pathogen by adult citrus psyllid reared on Citrus reticulata cv. Tank an infected with Candidatus Liberibacter as iatic us (CLas) , and the transmission efficiency to Citrus reticulata cv, Subcompress via CLas-positive psyllids. The results demonstrated that the two

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

    Science.gov (United States)

    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

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

    Directory of Open Access Journals (Sweden)

    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

  12. 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.

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

    Directory of Open Access Journals (Sweden)

    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

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

    Directory of Open Access Journals (Sweden)

    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

  15. Botanicals, selective insecticides, and predators to control Diaphorina citri (Hemiptera: Liviidae) in citrus orchards.

    Science.gov (United States)

    Khan, Azhar A; Afzal, Muhammad; Qureshi, Jawwad A; Khan, Arif M; Raza, Abubakar M

    2014-12-01

    The Asian citrus psyllid (ACP) Diaphorina citri Kuwayama vectors pathogens that cause huanglongbing (HLB) or citrus greening devastating and economically important disease present in most citrus growing regions. Young citrus shoots are required for psyllid reproduction and development. During winter citrus trees produce little or no new growth. Overwintering adults reproduce in spring on newly emerging shoots also attractive to other pests and beneficial insects. Botanicals and relatively selective insecticides could help to conserve beneficial insects and reduce pest resistance to insecticides. Sprays of Azadirachtin (Neem), Tropane (Datura), Spirotetramat, Spinetoram, and broad-spectrum Imidacloprid were evaluated to control ACP in spring and summer on 10-year-old "Kinow" Citrus reticulata Blanco trees producing new growth. Psyllid populations were high averaging 5-9 nymphs or adults per sample before treatment application. Nymphs or adults were significantly reduced to 0.5-1.5 per sample in all treatments for 3 weeks, average 61%-83% reduction. No significant reduction in ladybeetles Adalia bipunctata, Aneglei scardoni, Cheilomenes sexmaculata, and Coccinella septempunctata was observed. Syrphids, spiders and green lacewings were reduced in treated trees except with Tropane. Studies are warranted to assess impact of these predators on ACP and interaction with insecticides. Observed reduction in ACP populations may not be enough considering its reproductive potential and role in the spread of HLB. Follow-up sprays may be required to achieve additional suppression using rotations of different insecticides. © 2014 Institute of Zoology, Chinese Academy of Sciences.

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

    Science.gov (United States)

    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.

  17. 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.

  18. Detection of 'Candidatus Liberibacter asiaticus' in Diaphorina citri and its importance in the management of citrus huanglongbing in Florida.

    Science.gov (United States)

    Manjunath, K L; Halbert, S E; Ramadugu, C; Webb, S; Lee, R F

    2008-04-01

    Citrus huanglongbing (HLB or citrus greening), is a highly destructive disease that has been spreading in both Florida and Brazil. Its psyllid vector, Diaphorina citri Kuwayama, has spread to Texas and Mexico, thus threatening the future of citrus production elsewhere in mainland North America. Even though sensitive diagnostic methods have been developed for detection of the causal organisms, Candidatus Liberibacter spp., the pathogen cannot be detected consistently in plants until symptoms develop, presumably because of low titer and uneven distribution of the causal bacteria in nonsymptomatic tissues. In the present study, TaqMan based real-time quantitative polymerase chain reaction methodology was developed for detection of 'Ca. L. asiaticus' in D. citri. Over 1,200 samples of psyllid adults and nymphs, collected from various locations in Florida, from visually healthy and HLB symptomatic trees at different times of the year were analyzed to monitor the incidence and spread of HLB. The results showed that spread of 'Ca. L. asiaticus' in an area may be detected one to several years before the development of HLB symptoms in plants. The study suggests that discount garden centers and retail nurseries may have played a significant role in the widespread distribution of psyllids and plants carrying HLB pathogens in Florida.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

    2017-01-01

    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. PMID:28386418

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

    Science.gov (United States)

    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...

  2. 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.

  3. 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.

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

    Science.gov (United States)

    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.

  5. 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).

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  7. 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.

  8. Detection of citrus huanglongbing-associated 'Candidatus Liberibacter asiaticus' in citrus and Diaphorina citri in Pakistan, seasonal variability, and implications for disease management.

    Science.gov (United States)

    Razi, Muhammad F; Keremane, Manjunath L; Ramadugu, Chandrika; Roose, Mikeal; Khan, Iqrar A; Lee, Richard F

    2014-03-01

    We report the detection of the huanglongbing (HLB)-associated bacterium 'Candidatus Liberibacter asiaticus' from both plants and insects in Pakistan and the seasonal variability in the numbers of 'Ca. L. asiaticus'-positive psyllid vector, Diaphorina citri. Our studies showed that 'Ca. L. asiaticus' was detectable from trees in areas with maximum temperatures reaching nearly 50°C (average maximum of 42°C). However, the bacterium was present at very low levels in psyllids both in summer (June to August) and autumn (September to November) in contrast to reports from Florida, where the bacterium was detectable at very high levels during October to November. We hypothesize that hot summer temperatures in Pakistan may interfere with acquisition and replication of 'Ca. L. asiaticus' in psyllids and may lead to dead or non transmissible 'Ca. L. asiaticus' in plants. Psyllid counts were very low in both summer and winter, showed a population peak ('Ca. L. asiaticus'-positive vectors) in spring, and showed a larger peak ('Ca. L. asiaticus'-free psyllids) in autumn. Natural thermotherapy during hot summers and a low vector population during environmental extremes may have played a major role in long-term survival of the citrus industry in Pakistan. These results may be useful in developing management strategies for U.S. citrus industries in Texas and California.

  9. Roles of olfactory cues, visual cues, and mating status in orientation of Diaphorina citri Kuwayama (Hemiptera: Psyllidae) to four different host plants.

    Science.gov (United States)

    Wenninger, Erik J; Stelinski, Lukasz L; Hall, David G

    2009-02-01

    Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is an important worldwide pest of citrus that vectors bacteria (Candidatus Liberibacter spp.) responsible for huanglongbing (citrus greening disease). We examined the behavioral responses of mated and unmated D. citri of both sexes to odors from host plants in a Y-tube olfactometer, with and without visual cues. The host plants tested were 'Duncan' grapefruit (Citrus paradisi Macfayden), sour orange (Citrus aurantium L.), navel orange (C. sinensis L.), and Murraya paniculata L. Jack. Responses varied by plant species, psyllid sex and mating status, and the presence of a visual cue. Evidence of attraction generally was stronger in females and in mated individuals of both sexes relative to virgins. The presence of a visual cue typically enhanced attractiveness of olfactory cues; in no case did unmated individuals show evidence of attraction to host plant odors in the absence of avisual cue. In the absence of visual cues, mated females and males showed evidence of attraction only to odors from sour orange and navel orange, respectively. Psyllids exhibited anemotactic responses when assayed with plant odors alone but showed strong evidence of attraction only when olfactory and visual cues were combined, suggesting that olfactory cues facilitate orientation to host plants but may be insufficient alone. Antennal responses to citrus volatiles were confirmed by electroantennogram. The results reported here provide evidence that D. citri uses olfactory and visual cues in orientation to host plants and suggest the possibility of using plant volatiles in monitoring and management of this pest.

  10. Diaphorina citri (Hemiptera: Liviidae) responses to microcontroller-buzzer communication signals of potential use in vibration traps

    Science.gov (United States)

    Monitoring of Diaphorina citri Kuwayama populations is an important component of efforts to reduce damage caused by huanglongbing, a devastating disease it vectors in citrus groves. Currently, D. citri is monitored primarily by unbaited sticky traps or visual inspection of trees. A potentially more ...

  11. 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.

  12. 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.

  13. 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.

    Science.gov (United States)

    Leong, Stephen Chan Teck; Abang, Fatimah; Beattie, Andrew; Kueh, Roland Jui Heng; Wong, Sing King

    2012-01-01

    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 nC(24)HMO and conventional pesticides on the citrus psyllid population and parasitoids in citrus orchard were also determined.

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

    Science.gov (United States)

    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. PMID:28083006

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

    Science.gov (United States)

    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.

  16. Daily and seasonal patterns in abdomen color in Diaphoria citri (Hemiptera: Psyllidae)

    Science.gov (United States)

    Diaphorina citri Kuwayama, a psyllid vector of huanglongbing (citrus greening disease), exhibits three more or less distinct abdomen colors in the adult psyllid: gray/brown, blue/green, and orange/yellow. We explored the daily (in individuals in the laboratory) and seasonal (in a field population) p...

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

    Science.gov (United States)

    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 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.

  18. Asian citrus psyllid RNAi pathway : RNAi evidence

    OpenAIRE

    Taning, Clauvis N. T.; Andrade, Eduardo C.; Hunter, Wayne B.; Olivier Christiaens; Guy Smagghe

    2016-01-01

    Diaphorina citri, known as the Asian citrus psyllid, is an important pest of citrus because it transmits a phloem-limited bacteria strongly implicated in huanglongbing (citrus greening disease). Emerging biotechnologies, such as RNA interference, could provide a new sustainable and environmentally friendly strategy for the management of this pest. In this study, genome and functional analysis were performed to verify whether the RNAi core genes are present in the Asian psyllid genome and if t...

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

    Science.gov (United States)

    The Asian citrus psyllid (ACP), due to its potential to vector the pathogen causing citrus greening disease or huanglongbing, is one of the most serious citrus pests worldwide. While optimal plant cultivars for ACP oviposition and development have been determined, little is known of the influence of...

  20. Attraction of Diaphorina citri (Hemiptera: Liviidae) to colored light

    Science.gov (United States)

    The most devasting disease of citrus, huanglongbing (HLB), is spread by the insect vector the Asian Citrus Psyllid (ACP). Although ACP are known to respond to visual cues, previous studies have not documented the specific wavelengths of light attractive to ACP. The objectives of this study were to...

  1. 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

    Directory of Open Access Journals (Sweden)

    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

  2. Localized autoinoculation and dissemination of Isaria fumosorosea for control of the Asian citrus psyllid in South Texas

    Science.gov (United States)

    The Asian citrus psyllid, Diaphorina citri Kuwayama, vectors the causal organism of citrus greening disease. Integrated strategies are needed to control D. citri in south Texas. Control approaches involving entomopathogenic fungi may be useful on ornamental and abandoned citrus and other rutaceous...

  3. Asian Citrus Psyllid RNAi Pathway – RNAi evidence

    OpenAIRE

    Taning, Clauvis N. T.; Eduardo C. Andrade; Hunter, Wayne B.; Olivier Christiaens; Guy Smagghe

    2016-01-01

    Diaphorina citri, known as the Asian citrus psyllid, is an important pest of citrus because it transmits a phloem-limited bacteria strongly implicated in huanglongbing (citrus greening disease). Emerging biotechnologies, such as RNA interference, could provide a new sustainable and environmentally friendly strategy for the management of this pest. In this study, genome and functional analysis were performed to verify whether the RNAi core genes are present in the Asian psyllid genome and if t...

  4. Asian Citrus Psyllid RNAi Pathway – RNAi evidence

    OpenAIRE

    Taning, Clauvis N. T.; Andrade, Eduardo C. de; Hunter, Wayne B.; Christiaens, Olivier; Smagghe, Guy

    2016-01-01

    Diaphorina citri, known as the Asian citrus psyllid, is an important pest of citrus because it transmits a phloem-limited bacteria strongly implicated in huanglongbing (citrus greening disease). Emerging biotechnologies, such as RNA interference, could provide a new sustainable and environmentally friendly strategy for the management of this pest. In this study, genome and functional analysis were performed to verify whether the RNAi core genes are present in the Asian psyllid genome and if t...

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

    Science.gov (United States)

    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.

  6. Susceptibilidad de Diaphorina citri Kuwayama (Hemíptera : Liviidae) a insecticidas en Veracruz, México.

    OpenAIRE

    García Méndez, Víctor Hugo

    2013-01-01

    El uso racional de insecticidas requiere una evaluación previa de la efectividad de los productos para evitar aplicaciones inefectivas o el desarrollo de resistencia. El Psílido Asiático de los Cítricos (PAC), Diaphorina citri es vector del Huanglongbing y requiere estrategias de manejo regional de la resistencia. El objetivo de la investigación fue definir las líneas base de manejo de la resistencia y determinar la susceptibilidad de D. citri a insecticidas convencionales de uso común en aé...

  7. 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...

  8. 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...

  9. 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...

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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...

  13. Candidatus Liberibacter asiaticus is present in orange jasmine and Asian citrus psyllid reared from jasmine at low titers

    Science.gov (United States)

    Orange jasmine, Murraya paniculata, is a common horticultural plant in Florida, and an alternate host of the Asian citrus psyllid, Diaphorina citri Kuwayama. Orange jasmine has also been reported to harbor the bacteria Candidatus Liberibacter asiaticus, the causal agent of huanglongbing disease. We ...

  14. 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

    Science.gov (United States)

    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...

  15. Bionomics of Asian citrus psyllid (Hemiptera: Liviidae) associated with orange jasmine hedges in southest central Florida, with special reference to biological control by Tamarixia radiata

    Science.gov (United States)

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is an important pest in Florida because it vectors bacteria responsible for citrus huanglongbing disease. In addition to infesting citrus, orange jasmine (Murraya paniculata) is one of ACP’s favorite host plants and is widely grown as an orn...

  16. Retracted stylets in nymphs of the Asian citrus psyllid are held externally against the clypeus by a special paired organ not found in the adults

    Science.gov (United States)

    Differences in ultrastructure of the mouthparts in nymphs and adults of the Asian citrus psyllid Diaphorina citri (Hemiptera, Psyllidae), vector of the bacterium associated with citrus huanglongbing disease, were studied using scanning electron microscopy. The number of sensilla on the labial tip in...

  17. 柑橘木虱对柑橘不同品种的趋性%Diaphorina citri's Gost Preference on Thirteen Citrus Varieties

    Institute of Scientific and Technical Information of China (English)

    陈建利; 阮传清; 刘波; 范国成; 段永平; D.HALL

    2011-01-01

    用2种方法测定了柑橘木虱Diaphorina citri Kuw.成虫对13个柑橘品种离体嫩梢的选择性.结果表明,对不同品种进行两两组合的选择性试验,柑橘木虱停靠在处理组合早金和四季斑叶橘的虫量占总虫量的23%和1%,停靠在处理组合台湾玫瑰橙和纽荷尔脐橙上的虫量分别占总虫量的33%和20%,停靠在处理组合日辉和佩奇甜橙上虫量分别占总虫量的38%和15%,两两之间差异显著.将13个品种组合在一起进行选择性试验,结果表明柑橘木虱停靠在纽荷尔、福橘和佩奇甜橙上的成虫数量为每梢3.2~6.6只·d-1,显著高于木虱在其余10个品种枝梢上的停靠数量.福橘是柑橘黄龙病高发品种的原因是其对柑橘木虱的吸引力较强.%Under laboratory conditions, preference of adult Asian Citrus Psyllid (ACP), Diaphorina citri Kuw, for infestation among 13 citrus varieties was determined with two experimentations. Experiment A applied every 2 detached shoots of different citrus varieties and put together for choice. Experiment B used the detached shoots of all tested citrus varieties and put together for choice. The results of Experiment A revealed that the percentage of ACP adults on each shoot of Citrus sinensis (L.) Osbeck cv. Earlygold and C. medurensis (C. mitis) cv. Varigated Calamondin were 23 % and 1 %, respectively, which was significantly different from each other. The percentage on C. sinensis Osbeck cv. Taiwanmeiguicheng (33%) and cv. Newhall Navel Orange (20%) were also significantly different from each other. Significant different percentages of ACP adults were also observed between C. reticulata C. paradisi cv. Rihui (38%) and C. reticulata C. grandis cv. Page (15%). In Experiment B, the number of ACP on the shoots of C. sinensis Osbeck cv. Newhall Navel Orange, C. tangerina Tanaka cv. Fuju and C. reticulata Citrusgrandis cv. Page was 3.2-6.6 adults · shoot-day-1 , which were significantly higher than that

  18. Fluctuación poblacional de Diaphorina citri Kuwayana en limón persa (Citrus latifolia Tanaka) en la sabana de Huimanguillo, Tabasco.

    OpenAIRE

    García Garduza, Diógenes

    2012-01-01

    Diaphorina citri Kuwayama es el vector de la enfermedad Huanglongbing, la cual se encuentra actualmente en 14 de los 23 estados productores de cítricos en México. El estado de Tabasco, donde se cultivan 15, 406 ha de estos frutales, aún no es afectado por la enfermedad, por lo cual es importante tomar medidas para controlar las poblaciones del vector, cuya presencia en dicho estado fue detectada en 2005. El objetivo de este trabajo fue determinar la fluctuación poblacional de D. citri y la r...

  19. 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

    Directory of Open Access Journals (Sweden)

    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

  20. Wolbachia infection density in populations of the Asian citrus psyllid (Hemiptera: Liviidae).

    Science.gov (United States)

    Hoffmann, M; Coy, M R; Kingdom Gibbard, H N; Pelz-Stelinski, K S

    2014-10-01

    The symbiotic relationships between bacteria of the genus Wolbachia (order Rickettsiales) and their arthropod hosts are diverse and can range from mutualism to parasitism. Whereas effects of Wolbachia on host biology are well investigated, little is known about diversity and abundance of Wolbachia in their natural hosts. The phloem-feeding Asian citrus psyllid, Diaphorina citri (Kuwayama) (Hemiptera: Liviidae), is naturally infected with Wolbachia (wDi). In the current study, we calculated the within-host density of Wolbachia in Florida D. citri populations using quantitative polymerase chain reaction for detection of the Wolbachia outer surface protein gene, wsp. Gene quantities were normalized to the D. citri wingless gene (Wg) to estimate Wolbachia abundance in individual D. citri. Using this method, significant geographic differences in Wolbachia densities were detected among Florida D. citri populations, with higher infection levels occurring in male versus female hosts.

  1. Perspectives to the use of entomopathogenic fungi for biological control of Diaphorina citri in Mexico

    OpenAIRE

    Sánchez-Peña, S. R.; Guizar-Guzmán, L.; Torres-Acosta, R. I.; López-Arroyo, J. I.; Casique-Valdés, R.

    2014-01-01

    Entomopathogenic fungi are natural enemies of D. citri adults in Mexico. Natural prevalence (%) of fungi on live adults in central Veracruz (summer) and southern Tamaulipas (fall) were: Hirsutella citriformis (7-35); Isaria fumosorosea (5-15); Lecanicillium (2 in Tamaulipas), and Beauveria (

  2. Construction and evaluation of yeast two-hybrid cDNA library of Diaphorina citri (Hemiptera: Psyllidae)%柑橘木虱酵母双杂交cDNA文库的构建及评价

    Institute of Scientific and Technical Information of China (English)

    马晓芳; 陈国庆; 张学潮; 徐海君

    2013-01-01

    In order to study the interacting proteins between the Asian citrus psyllid ( Diaphorina citri) and Candidatus Liberibacter asiaticus ( CLas) which is the pathogenic bacterium causing Huanglongbing, yeast two-hybrid cDNA library of D. citri was constructed using the Switching Mechanism at 5' End of the RNA Transcript (SMART) technique. The total RNA was isolated from the citrus psyllid adults bred in the laboratory and subjected to reverse transcription, and the double-strand cDNAs ( ds cDNAs) were synthesized. The ds cDNAs were ligated with homologous adapter and purified by the chromatography column. By using homologous recombination reaction, the ds cDNAs were transformed into the Y187 competent cell with the library plasmid pGADT7-Rec to construct yeast two-hybrid cDNA library. Detection of the library indicated that it contained more than 106 independent clones, the titer of the amplified library was 2. 23 × 10 cfu/mL, and the average size of inserts was above 750 bp in the cDNA library. These results demonstrate that the library meets the requirements of the standard cDNA library. Moreover, two membrane proteins, ORF420 and ORF3420, from ( CLas) were used as bait proteins to screen the interacting proteins in the library, but no positive clone was screened in the tests. The yeast two-hybrid cDNA library of D. citri will be useful for the research on the interaction between insect vectors and C. Liberibacter asiaticus in the future.%为了探索研究柑橘木虱Diaphorina citri与柑橘黄龙病(Huanglongbing,HLB)病原菌的相互作用蛋白,本研究运用RNA转录中5 '末端转换机制(Switching Mechanism at 5'End of the RNA Transcript,SMART)技术构建了柑橘木虱的酵母双杂交cDNA文库.以实验室饲养的柑橘木虱为材料,提取总RNA,经反转录后合成ds cDNA,两端添加同源重组序列,并用层析柱纯化;ds cDNA与文库质粒pGADT7-Rec在酵母Y187感受态细胞内发生同源重组,柑橘木虱cDNA重组到文库质粒

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

    Directory of Open Access Journals (Sweden)

    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.

  4. Biology and management of Asian citrus psyllid, vector of the huanglongbing pathogens.

    Science.gov (United States)

    Grafton-Cardwell, Elizabeth E; Stelinski, Lukasz L; Stansly, Philip A

    2013-01-01

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the most important pest of citrus worldwide because it serves as a vector of "Candidatus Liberibacter" species (Alphaproteobacteria) that cause huanglongbing (citrus greening disease). All commercially cultivated citrus is susceptible and varieties tolerant to disease expression are not yet available. Onset of disease occurs following a long latent period after inoculation, and thus the pathogen can spread widely prior to detection. Detection of the pathogen in Brazil in 2004 and Florida in 2005 catalyzed a significant increase in research on D. citri biology. Chemical control is the primary management strategy currently employed, but recently documented decreases in susceptibility of D. citri to several insecticides illustrate the need for more sustainable tools. Herein, we discuss recent advances in the understanding of D. citri biology and behavior, pathogen transmission biology, biological control, and chemical control with respect to "Candidatus Liberibacter asiaticus." Our goal is to point toward integrated and biologically relevant management of this pathosystem.

  5. Asian Citrus Psyllid RNAi Pathway – RNAi evidence

    Science.gov (United States)

    Taning, Clauvis N. T.; Andrade, Eduardo C.; Hunter, Wayne B.; Christiaens, Olivier; Smagghe, Guy

    2016-01-01

    Diaphorina citri, known as the Asian citrus psyllid, is an important pest of citrus because it transmits a phloem-limited bacteria strongly implicated in huanglongbing (citrus greening disease). Emerging biotechnologies, such as RNA interference, could provide a new sustainable and environmentally friendly strategy for the management of this pest. In this study, genome and functional analysis were performed to verify whether the RNAi core genes are present in the Asian psyllid genome and if the RNAi machinery could be exploited to develop a management strategy for this pest. Analyses of RNAi-related genes in the Asian citrus psyllid genome showed an absence of sequences encoding R2D2, a dsRNA-binding protein that functions as a cofactor of Dicer-2 in Drosophila. Nevertheless, bioassays using an in Planta System showed that the Asian citrus psyllid was very sensitive to ingested dsRNA, demonstrating a strong RNAi response. A small dose of dsRNA administered through a citrus flush was enough to trigger the RNAi mechanism, causing significant suppression of the targeted transcript, and increased psyllid mortality. This study provides evidence of a functional RNAi machinery, which could be further exploited to develop RNAi based management strategies for the control of the Asian citrus psyllid. PMID:27901078

  6. Annotation of the Asian Citrus Psyllid Genome Reveals a Reduced Innate Immune System.

    Science.gov (United States)

    Arp, Alex P; Hunter, Wayne B; Pelz-Stelinski, Kirsten S

    2016-01-01

    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 Kuwayama (Hemiptera: Liviidae). Hemipterans characterized to date lack a number of insect immune genes, including those associated with the Imd pathway targeting Gram-negative bacteria. The D. citri draft genome was used to characterize the immune defense genes present in D. citri. Predicted mRNAs identified by screening the published D. citri annotated draft genome were manually searched using a custom database of immune genes from previously annotated insect genomes. Toll and JAK/STAT pathways, general defense genes Dual oxidase, Nitric oxide synthase, prophenoloxidase, and cellular immune defense genes were present in D. citri. In contrast, D. citri lacked genes for the Imd pathway, most antimicrobial peptides, 1,3-β-glucan recognition proteins (GNBPs), and complete peptidoglycan recognition proteins. These data suggest that D. citri has a reduced immune capability similar to that observed in A. pisum, P. humanus, and R. prolixus. The absence of immune system genes from the D. citri genome may facilitate CLas infections, and is possibly compensated for by their relationship with their microbial endosymbionts.

  7. 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.

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

    Science.gov (United States)

    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.

  9. 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.

  10. 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.

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

    Science.gov (United States)

    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.

  12. Asian citrus psyllids and shade: Survival of Diaphorina citri on Murraya exotica foliage exposed to different levels of light

    Science.gov (United States)

    Using shade cloths of different densities, potted individuals of Murraya exotica (Rutaceae) were exposed to levels of photosynthetic radiation (PAR) varying from 48 µmol photons per meter2 per second to 1562 µmol photons per meter2 per second (average of three readings taken at noon during condition...

  13. 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.

    Science.gov (United States)

    Ammar, El-Desouky; Ramos, John E; Hall, David G; Dawson, William O; Shatters, Robert G

    2016-01-01

    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 adults

  14. 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

  15. Control químico de Diaphorina citri Kuwayama (Hemíptera: Psyllidae en lima persa Citrus latifolia Tanaka Chemical control of Diaphorina citri Kuwayama (Hemiptera: Psyllidae in Persian lime Citrus latifolia Tanaka

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    Luis Martín Hernández-Fuentes

    2012-06-01

    Full Text Available A nivel mundial, la citricultura representa una actividad de gran importancia. México ocupa el cuarto lugar mundial en producción. Debido a la introducción y dispersión del vector D. citri en todas las zonas citrí colas de México y la reciente detección de la enfermedad conocida como huanglongbing causada por la bacteria Candidatus Liberibacter asiaticus, es necesaria la evaluación de métodos de control de este insecto. En el año 2010 en Nayarit, México, se realizaron pruebas de efectividad técnica en campo y persistencia de productos químicos de diferente mecanismo de acción contra D. citri en lima persa. Se hicieron dos evaluaciones en fechas distintas. En cada ensayo se evaluó número de ninfas por brote y porcentaje de brotes infestados. De los productos evaluados el dimetoato, imidacloprid y la mezcla de imidacloprid más betaciflutrina ejercieron más de 85 % de control de ninfas durante 27 días después de la aplicación. En los árboles tratados con dimetoato e imidacloprid en dosis de 400 mL y 300 mL ha-¹, respectivamente, se observó 100% de brotes libres de ninfas de D. citri a los 27 días después de la aplicación. El aceite agrícola en dosis de dos y tres L ha-1 ejerció menor control de ninfas de D. citri.Citrus crops represent an outstanding activity at worldwide level. Mexico ranks fourth place in production. Due introduction and dispersion of D. citri vector in all citric zones from Mexico and the newly detected disease known as huanglongbing (HLB caused by Candidatus Liberibacter asiaticus bacteria, assessment of control methods for this bug is required. In 2010, technical effectiveness in field and persistence for different action route against Persian lime D. citri chemical products tests were performed in Nayarit, Mexico. Two assessments were made at different dates. The amount of nymphs per shoot and percentage of infested shoots were assessed in each rehearsal. From evaluated products, dimethoate

  16. 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.

  17. Control químico de Diaphorina citri Kuwayama (Hemíptera: Psyllidae) en lima persa Citrus latifolia Tanaka Chemical control of Diaphorina citri Kuwayama (Hemiptera: Psyllidae) in Persian lime Citrus latifolia Tanaka

    OpenAIRE

    Luis Martín Hernández-Fuentes; Mario Alfonso Urias-López; José Isabel López-Arroyo; Rafael Gómez-Jaimes; Néstor Bautista-Martínez

    2012-01-01

    A nivel mundial, la citricultura representa una actividad de gran importancia. México ocupa el cuarto lugar mundial en producción. Debido a la introducción y dispersión del vector D. citri en todas las zonas citrí colas de México y la reciente detección de la enfermedad conocida como huanglongbing causada por la bacteria Candidatus Liberibacter asiaticus, es necesaria la evaluación de métodos de control de este insecto. En el año 2010 en Nayarit, México, se realizaron pruebas de efectividad t...

  18. Effect of Cyantraniliprole, a Novel Insecticide, on the Inoculation of Candidatus Liberibacter Asiaticus Associated with Citrus Huanglongbing by the Asian Citrus Psyllid (Hemiptera: Liviidae).

    Science.gov (United States)

    Ammar, El-Desouky; Hall, David G; Alvarez, Juan M

    2015-04-01

    The Asian citrus psyllid (Diaphorina citri Kuwayama) is the principal vector of 'Candidatus Liberibacter asiaticus' (CLas) associated with huanglongbing (HLB), the most serious citrus disease worldwide. New control measures including pesticides are urgently needed to combat HLB, especially to protect young or newly planted citrus trees from CLas-inoculation by vector psyllids. Here, we tested CLas-inoculation by D. citri adults (CLas-exposed, reared on infected plants) by feeding them for 7 d on excised healthy citrus leaves with dry residues of cyantraniliprole (Exirel), a novel insecticide, in comparison with fenpropathrin (Danitol 2.4EC), an insecticide commonly used against D. citri. Fewer adults settled (putatively feeding or probing) on leaves treated with cyantraniliprole than those treated with fenpropathrin or water controls. Also, psyllid adults died at a slower rate on leaves treated with cyantraniliprole than those treated with fenpropathrin, although the final cumulative mortality did not differ between the two treatments. In quantitative real-time polymerase chain reaction tests, 59.0-65.3% of the CLas-exposed psyllid adults were proven to be CLas-positive. Inoculation rates of CLas (using 10 adults per leaf) into untreated healthy citrus leaves (47.5-85%) were significantly higher than rates into leaves treated with cyantraniliprole or fenpropathrin (2.5-12.5%). Reduced inoculation rates to leaves treated with cyantraniliprole probably occurred as a result of reduced feeding or probing by D. citri. The excised leaf assay method, which took only a few weeks compared with up to a year or longer using whole plants, can be an effective tool for testing the effect of new pesticides or other treatments in reducing CLas inoculation or transmission by psyllid vectors. Published by Oxford University Press on behalf of Entomological Society of America 2015. This work is written by a US Government employees and is in the public domain in the US.

  19. The push-pull strategy for citrus psyllid control.

    Science.gov (United States)

    Yan, Huaxue; Zeng, Jiwu; Zhong, Guangyan

    2015-07-01

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the only natural vector of Candidatus Liberibacter asiaticus that causes citrus huanglongbing (HLB), a most destructive disease of citrus. Currently, no remedial therapy exists for the disease, and so effective control of ACP is very important in curbing the transmission of the disease. The push-pull strategy should be thoroughly explored as an approach to ACP management. This mini-review summarises the current progress towards more effective repellent and attractant chemicals through investigating known repellent and attractive plants. Interactions between ACP and its host plants are also addressed, with emphasis on the possible involvement of the host biochemicals in attracting the insect. Potential ways to increase the effectiveness of the pull-push strategy are briefly discussed. It is expected that the pull-push strategy will be gradually developed following more extensive research.

  20. Insecticide resistance of adults and nymphs of Asian citrus psyllid populations from Apatzingán Valley, Mexico.

    Science.gov (United States)

    Pardo, Saúl; Martínez, Ana M; Figueroa, José I; Chavarrieta, Juan M; Viñuela, Elisa; Rebollar-Alviter, Ángel; Miranda, Mario A; Valle, Javier; Pineda, Samuel

    2017-07-18

    Control of the Asian citrus psyllid Diaphorina citri Kuwayama, the most important pest of citrus worldwide, is based on the use of insecticides, though unsatisfactory results have recently been reported. In this study, insecticide resistance of D. citri to three insecticides (bifenthrin, malathion, and chlorpyrifos) was examined. Three populations (designated Dci-CParácuaro, Dci-El Junco, and Dci-Antúnez) of both adults and fourth-instar D. citri individuals were collected in 2014 at two different times and on one occasion, respectively, from three locations (Crucero de Parácuaro, El Junco, and Antúnez). These locations represent the major commercial Mexican lemon production areas in the Apatzingán Valley in the state of Michoacán, Mexico. The three populations of D. citri adults and fourth-instar nymphs at the different collection times showed low levels of resistance (≤7-fold) to bifenthrin, but were very resistant to malathion (≤345- and ≤432-fold for adults and fourth instars, respectively) and chlorpyrifos (≤2435- and ≤1424-fold for adults and fourth instars, respectively). Resistance levels to the tested insecticides were highly variable but homogeneous among seasons and localities. Resistance management programmes that include crop sanitation, use of biological and cultural control practices, and rotation of insecticide classes should be established, particularly in areas where D. citri has developed resistance to malathion and chlorpyrifos. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  1. 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.

  2. Association of ‘Candidatus Liberibacter asiaticus’, the causal agent of citrus huanglongbing in Murraya paniculata and Diaphorina citri in Thailand

    Science.gov (United States)

    Orange jasmine, Murraya paniculata, is a preferred alternative host for the Asian citrus psyllid, the primary vector of citrus Huanglongbing (HLB) disease caused by ‘Candidatus Liberibacter asiaticus’ (Las). M. paniculata plant samples and psyllids on the Murraya plants from ten diverse geographical...

  3. 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; Lapointe, Stephen L

    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

  4. Odor coding in a disease-transmitting herbivorous insect, the Asian citrus psyllid.

    Science.gov (United States)

    Coutinho-Abreu, Iliano V; McInally, Shane; Forster, Lisa; Luck, Robert; Ray, Anandasankar

    2014-07-01

    Olfactory systems discriminate odorants very efficiently and herbivorous insects use them to find hosts in confounding and complex odor landscapes. The Asian citrus psyllid (ACP), Diaphorina citri, feeds on citrus flush and transmits Candidatus Liberibacter that causes citrus greening disease globally. Here, we perform a systematic analysis of odor detection in the ACP antenna using single-unit electrophysiology of rhinarial plate sensilla to a large panel of odorants from plants. We identify neurons that respond strongly to odorants found in the host citrus plants. Comparisons with the generalist yeast-feeding Drosophila melanogaster and specialist anthropophilic Anopheles gambiae reveal differences in odor-coding strategies for the citrus-seeking ACP. These findings provide a foundation for understanding host-odor coding in herbivorous insects.

  5. Monitoring for Insecticide Resistance in Asian Citrus Psyllid (Hemiptera: Psyllidae) Populations in Florida.

    Science.gov (United States)

    Kanga, Lambert H B; Eason, Julius; Haseeb, Muhammad; Qureshi, Jawwad; Stansly, Philip

    2016-04-01

    The development of insecticide resistance in Asian citrus psyllid, Diaphorina citri Kuwayama, populations is a serious threat to the citrus industry. As a contribution to a resistance management strategy, we developed a glass vial technique to monitor field populations of Asian citrus psyllid for insecticide resistance. Diagnostic concentrations needed to separate susceptible genotypes from resistant individuals were determined for cypermethrin (0.5 μg per vial), malathion (1.0 μg per vial), diazinon (1.0 μg per vial), carbaryl (1.0 μg per vial), carbofuran (0.1 μg per vial), methomyl (1.0 μg per vial), propoxur (1.0 μg per vial), endosulfan (1.0 μg per vial), imidacloprid (0.5 μg per vial), acetamiprid (5.0 μg per vial), chlorfenapyr (2.5 μg per vial), and fenpyroximate (2.5 μg per vial). In 2014, resistance to two carbamate insecticides (carbaryl and carbofuran), one organophosphate (malathion), one pyrethroid (cypermethrin), and one pyrazole (fenpyroximate) was detected in field populations of Asian citrus psyllid in Immokalee, FL. There was no resistance detected to diazinon, methomyl, propoxur, endosulfan, imidacloprid, and chlorfenapyr. The levels of insecticide resistance were variable and unstable, suggesting that resistance could be successfully managed. The results validate the use of the glass vial bioassay to monitor for resistance in Asian citrus psyllid populations and provide the basis for the development of a resistance management strategy designed to extend the efficacy of all classes of insecticides used for control of the Asian citrus psyllid.

  6. New excised-leaf assay method to test inoculativity of Asian citrus psyllid (Hemiptera: Psyllidae) with Candidatus Liberibacter asiaticus associated with citrus huanglongbing disease.

    Science.gov (United States)

    Ammar, El-Desouky; Walter, Abigail J; Hall, David G

    2013-02-01

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the primary vector of Candidatus Liberibacter asiaticus (Las) associated with huanglongbing, or citrus greening, the most devastating citrus (Citrus spp.) disease worldwide. Here, we developed a new "excised-leaf assay" that can speed up Las-inoculativity tests on Asian citrus psyllid from the current 3-12 mo (when using whole citrus seedlings for inoculation) to only 2-3 wk. Young adults of Asian citrus psyllid that had been reared on Las-infected plants were caged on excised healthy sweet orange [Citrus sinensis (L.) Osbeck] leaves for a 1-2-wk inoculation access periods (IAP), and then both psyllids and leaves were tested later by quantitative polymerase chain reaction (PCR). When single adults were tested per leaf, percentages of Las-positive leaves averaged 2-6% by using HLBaspr primers and 10-20% by using the more sensitive LJ900 primers. Higher proportions of Las-positive leaves were obtained with 1) higher densities of inoculating psyllids (5-10 adults per leaf), 2) longer IAPs, and 3) incubation of leaves for 1 wk postinoculation before PCR. Logistic regression analysis indicated a positive correlation between Las titer in Asian citrus psyllid adults tested singly and the probability of detecting Las in the inoculated leaves, correlations that can be very useful in epidemiological studies. Comparison between excised leaves and whole seedlings, inoculated consecutively for 1 wk each by one or a group of psyllids, indicated no significant difference between Las detection in excised leaves or whole plants. This new excised-leaf assay method saves considerable time, materials, and greenhouse space, and it may enhance vector relation and epidemiological studies on Las and potentially other Liberibacter spp. associated with huanglongbing disease.

  7. 联苯菊酯等6种农药对柑桔木虱成虫室内毒力测定及田间药效试验%Field Control Experiments and Toxicity of 6 Pesticides to Asian Citrus Psylla(Diaphorina citri Kuwayama)

    Institute of Scientific and Technical Information of China (English)

    邓明学; 谭有龙; 唐美欣; 陈贵峰; 唐明丽

    2009-01-01

    Citrus psylla (Diaphorina citri) has become the most important pest in Huanglongbing (HLB) areas. Up to now; the assay was yet not reported. By means of dipping, the laboratory toxicity of 6 pesticides to adult psylla was conducted out in 2008. The LC of carbosulfan, phenthoate,bifenthrin,imidacloprid, quinalphos,Piri-miphos-methyl was 4.1mg/kg, 44.3 mg/kg, 44.3 mg/kg, 60.1 mg/kg, 73.5 mg/kg, 192.8 mg/kg, respectively. Af-ter treating of 24 h, 3d and 7 d, the adult psyUid field control efficiency of bifenthrin plus azone 12.5 mg (a.i)/kg was all 100%. the adult psyllid control efficiency of phenthoate 333 mg(a.i)/kg,quinalphos 250 mg(a.i)/kg, carbo-suffan plus azone 200(a.i)/kg was all over than 95%, while that of pirimiphos-methyl 500 mg(a.i)/kg less 80%. The adult psyllid field control efficiency of carbosulfan plus azone 200(a.i)/kg was better than that of carbosul-fan 250(a.i)/kg.%为了筛选到更多防治柑桔木虱成虫的有效药剂,于2008年采用点滴法测定了6种农药对其成虫的室内毒力,活性最高的是丁硫克百威(LC50=4.1 mg/kg),其次分别是稻丰散(LC50=44.3 mg/kg)、联苯菊酯(LC50=44.3 mg/kg)、吡虫啉(LC50=60.1 mg/kg)和喹硫磷(LC50=73.5mg/kg),最差的是甲基嘧啶磷(LC50=192.8mg/kg).田间药效试验结果表明,增效联苯菊酯EC 25.00,16.67,12.50 mg(a.i)/kg药后24 h,3,7天的防效达100%;喹硫磷EC 250 mg(a.i)/kg、稻丰散EC 500 mg(a.i)/kg、333 mg(a.i)/kg、增效丁硫克百威EW200 mg(a.i)/kg药后24 h的防效>95%;甲基嘧啶磷500 mg(a.i)/kg的防效低于80%.增效丁硫克百威EW200mg(a.i)/kg的防效高于丁硫克百威EC 250mg(a.i)/kg的防效.

  8. Male Psyllids Differentially Learn in the Context of Copulation

    Directory of Open Access Journals (Sweden)

    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.

  9. Male Psyllids Differentially Learn in the Context of Copulation

    Science.gov (United States)

    Stockton, Dara G.; Martini, Xavier; Stelinski, Lukasz L.

    2017-01-01

    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. PMID:28178203

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    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...

  12. Use of micro-CT to elucidate details of the anatomy and feeding of the Asian citrus psyllid Diaphorina citri Kuwayama, 1908 (Insecta: Hemiptera, Liviidae)

    Science.gov (United States)

    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...

  13. Diversity of the citrus HLB bacterium, ‘Candidatus Liberibacter asiaticus’, in psyllids (Diaphorina citri) collected from Murraya paniculata and citrus spp. in Thailand

    Science.gov (United States)

    ‘Candidatus Liberibacter asiaticus’ (Las) is a phloem inhabiting bacterium that causes huanglongbing disease (HLB), also known as citrus greening associated with three species of a-Proteobacteria in the genus ‘Candidatus Liberibacter sp’. Prophage is an important genetic element of bacterial genomes...

  14. Wavelength and polarization affect phototaxis of the Asian citrus psyllid

    Science.gov (United States)

    The Asian citrus psyllid, D. citri, is a primary pest for citrus production due to its status as a vector of the citrus disease, huanglongbing. We investigated phototactic behavior of D. citri to evaluate effects of light of specific wavelength or polarization using a horizontal bioassay arena. Wave...

  15. Transmission Rates of ‘Ca. Liberibacter asiaticus’ by Asian Citrus Psyllid Are Enhanced by the Presence and Developmental Stage of Citrus Flush.

    Science.gov (United States)

    Hall, David G; Albrecht, Ute; Bowman, Kim D

    2016-04-01

    Asian citrus psyllid (Diaphorina citri Kuwayama) transmits a bacterium ‘Candidatus Liberibacter asiaticus’ (CLas) putatively responsible for a devastating citrus disease known as Asiatic huanglongbing (HLB) (citrus greening disease). The psyllid and disease have invaded many citrus-growing regions including the United States, where the disease is seriously jeopardizing the Florida citrus industry. We recently concluded research that showed CLas transmission rates are increased when citrus flush is present. Flush is any new leaf growth ranging in development from first emergence up until the leaves are fully expanded yet still tender. In an experiment with seedlings of a rootstock cultivar ‘US-942’, a 1-wk infestation of 20 Asian citrus psyllids from an infected colony resulted in 53–60% of seedlings becoming infected when flush was present compared with only 7% when no flush was present. In a second experiment with ‘US-942’, 77–97% of seedlings became infected when flush was present compared with 40% when no flush was present. A similar experiment with ‘Valencia’ sweet orange resulted in 23, 80, and 3% seedlings becoming infected when young, older, or no flush was present, respectively. Young plants are therefore more likely to contract HLB if flush is present, with older flush promoting higher infection rates under the conditions of this study. Based on this finding, healthy citrus should be protected from Asian citrus psyllid infestations throughout a flush. To evaluate germplasm for CLas resistance, inoculations using infected Asian citrus psyllid would best be achieved if flush is present.

  16. Asian Citrus Psyllid Expression Profiles Suggest Candidatus Liberibacter Asiaticus-Mediated Alteration of Adult Nutrition and Metabolism, and of Nymphal Development and Immunity.

    Science.gov (United States)

    Vyas, Meenal; Fisher, Tonja W; He, Ruifeng; Nelson, William; Yin, Guohua; Cicero, Joseph M; Willer, Mark; Kim, Ryan; Kramer, Robin; May, Greg A; Crow, John A; Soderlund, Carol A; Gang, David R; Brown, Judith K

    2015-01-01

    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 transmission.

  17. Asian Citrus Psyllid Expression Profiles Suggest Candidatus Liberibacter Asiaticus-Mediated Alteration of Adult Nutrition and Metabolism, and of Nymphal Development and Immunity.

    Directory of Open Access Journals (Sweden)

    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

  18. 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.

  19. Trailers transporting oranges to processing plants move Asian citrus psyllids

    Science.gov (United States)

    Huanglongbing (citrus greening) is one of the most serious of citrus diseases. Movement of the disease occurs as a result of natural vector-borne infection and by movement of plant material. We demonstrate here that Diaphorina citri Kuwayama (vector of citrus greening pathogens) can be transported i...

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

    Science.gov (United States)

    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.

  1. The relevance of Murraya paniculata and related species as potential hosts and inoculum reservoirs of Candidatus Liberibacter asiaticus, causal agent of Huanglongbing (HLB)

    Science.gov (United States)

    Huanglongbing (HLB), considered the most serious insect-vectored bacterial disease of citrus, is transmitted in nature by the Asian citrus psyllid Diaphorina citri and the African citrus psyllid Trioza erytreae. Diaphorina citri was discovered in southern Florida in 1998 and the HLB disease in 2005...

  2. Chemical control of the Asian citrus psyllid and of huanglongbing disease in citrus.

    Science.gov (United States)

    Boina, Dhana Raj; Bloomquist, Jeffrey R

    2015-06-01

    By 2014, huanglongbing (HLB), the most destructive disease of citrus, and its insect vector, the Asian citrus psyllid (ACP), Diaphorina citri (Kuwayama), became established in all major citrus-growing regions of the world, including the United States, with the exception of California. At present, application of insecticides is the most widely followed option for reducing ACP populations, while application of antibiotics for suppressing HLB disease/symptoms is being practiced in some citrus-growing regions. Application of insecticides during the dormant winter season, along with cultivation of HLB-free seedlings and early detection and removal of symptomatic and asymptomatic trees, has been very effective in managing ACP. Area-wide management of ACP by application of insecticides at low volume in large areas of citrus cultivation has been shown to be effective in managing HLB and reducing management costs. As insecticide resistance is a major problem in sustainable management of ACP, rotation/alternation of insecticides with different chemistries and modes of action needs to be followed. Besides control of the insect vector, use of antibiotics has temporarily suppressed the symptoms of HLB in diseased trees. Recent efforts to discover and screen existing as well as new compounds for their antibiotic and antimicrobial activities have identified some promising molecules for HLB control. There is an urgent need to find a sustainable solution to the HLB menace through chemical control of ACP populations and within HLB-infected trees through the judicious use of labeled insecticides (existing and novel chemistries) and antibiotics in area-wide management programs with due consideration to the insecticide resistance problem. © 2014 Society of Chemical Industry.

  3. 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.

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

    Science.gov (United States)

    Kruse, Angela; Fattah-Hosseini, Somayeh; Saha, Surya; Johnson, Richard; Warwick, EricaRose; Sturgeon, Kasie; Mueller, Lukas; MacCoss, Michael J; Shatters, Robert G; Cilia Heck, Michelle

    2017-01-01

    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.

  5. Economic injury levels for Asian citrus psyllid control in process oranges from mature trees with high incidence of huanglongbing.

    Science.gov (United States)

    Monzo, Cesar; Stansly, Philip A

    2017-01-01

    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 maintain economic

  6. Formic and acetic acids in degradation products of plant volatiles elicit olfactory and behavorial responses from an insect vector

    Science.gov (United States)

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae) vectors the bacterial pathogen presumed to be the etiological agent of citrus greening disease, Huanglongbing (HLB), a major threat to citrus industry worldwide. We studied antennal and behavioral responses of Diaphorina citri...

  7. Murraya paniculata (orange jasmine), a host and possible inoculum reservoir for Candidatus Liberibacter asiaticus, causal agent of Huanglongbing

    Science.gov (United States)

    Huanglongbing (HLB), considered the most serious vectored-pathogen of citrus, is transmitted in nature by the Asian citrus psyllid Diaphorina citri and the African citrus psyllid Trioza erytreae. In 1999, D. citri was discovered in southern Florida and has become established in FL and TX. Huanglon...

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

    Science.gov (United States)

    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...

  9. PROCENTAJE DE PSILIDOS PORTADORES DE Canidatus Liberibacter asiaticus Y SU RELACION CON LA INCIDENCIA DE HLB EN CINCO HUERTAS COMERCIALES DE LIMON MEXICANO {Citrus aurantifolia (Christm) Swingle} EN EL ESTADO DE COLIMA MEXICO

    Science.gov (United States)

    Asian citrus psyllids (Diaphorina citri) were individually analyzed by qPCR to detect Candidatus Liberibacter asiaticus (CLas). The psyllids were collected in Mexican lime (Citrus aurantifolia) trees in five commercial orchards in Tecomán and Manzanillo, Colima with severe symptoms of classical mott...

  10. Microbial diversity in Diaphorina citri( Homoptera: Psyllidae) estimated by 16S rDNA analysis using DGGE and RFLP%基于16S rDNA序列的柑桔木虱体内共生菌多样性研究

    Institute of Scientific and Technical Information of China (English)

    殷幼平; 刘婷婷; 田圣超; 胡修峰; 吴东; 王中康

    2011-01-01

    昆虫消化道内是一个复杂的微生态系统,有大量的微生物存在.这些微生物对寄主发育、营养吸收和防御方面都起着重要的作用.本文利用基于16S rRNA基因的PCR-RFLP指纹图谱法和变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)的方法对柑桔黄龙病虫-媒柑桔木虱Diaphorina citri体内细菌菌群多样性进行了研究.经PCR-RFLP分析显示31条序列与变形菌门的假单胞菌科、肠杆菌科、黄单胞菌科、伯克氏菌科、立克次氏菌科和根瘤菌科细菌具有较高同源性.柑桔木虱内共生细菌的优势菌群依次为合胞体共生菌(syncytiumendosymbiont)(同源性99%,5条序列,分离频率31%)、Candidatus Carsonella ruddii和Mycetocyte内共生菌(同源性98%,5条序列,分离频率31%)以及亚洲韧皮杆菌(Candidatus Liberibacter asiaticus)和内共生菌Wolbachia.对柑桔木虱内生细菌16S rDNA V3区序列的PCR-DGGE分析,条带相似性的UPMAGA聚类分析表明,采自九里香Murraya paniculata的柑桔木虱内生细菌大多聚为一支,而来自柑桔的聚在另一支,说明寄主差异对柑桔木虱内生细菌菌群构成的影响大于地理位置的影响.将PCR-DGGE条带测序,序列经GenBank序列比对发现柑桔木虱内生细菌主要属于变形菌门假单胞菌科、立克次氏菌科、肠杆菌科、黄单胞菌科以及厚壁菌门链球菌科和芽孢杆菌科.合胞体共生菌(条带3-4)因其稳定存在于木虱体内且不随柑桔木虱寄主和地理位置的改变而变化,可能是柑桔木虱体内的优势共生菌;内共生细菌Wolbachia也在柑桔木虱内稳定存在,表明我国柑桔木虱感染Wolbachia是普遍现象.PCR-RFLP和PCR-DGGE两种方法相结合较好地反映了柑桔木虱内生细菌菌群的多样性,而且均显示出柑桔木虱内的合胞体共生菌是主要的优势菌群.

  11. Visualization of ‘Candidatus Liberibacter asiaticus’ cells in citrus seed coats with fluorescence in situ hybridization and transmission electron microscopy

    Science.gov (United States)

    ‘Candidatus Liberibacter asiaticus’ is bacterium implicated as the causal agent of the economically damaging disease of citrus called huanglongbing (HLB). The bacterium is spread by movement of infected citrus propagation material and by the Asian citrus psyllid, Diaphorina citri Kuwayama. Seed tr...

  12. Broad sprectrum potential of Isaria fumosorosea on insect pests of citrus

    Science.gov (United States)

    Use of entomopathogenic fungi, Isaria fumosorosea, Ifr, =Paecilomyces fumosoroseus, successfully increased insect pest mortality. Spraying the Ifr containing product, PFR97 TM, on citrus seedlings was used to screen efficacy for the management of Asian citrus psyllid, Diaphorina citri; glassy-winge...

  13. 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.

  14. A comparison of plant species for rearing Asian citrus psyllid

    Science.gov (United States)

    Five plant genotypes were compared with respect to Asian citrus psyllid (ACP) reproduction potential: Bergera koenigii, Citrus aurantiifolia, C. macrophylla, C. taiwanica and Murraya paniculata. Asian citrus psyllid reproduction is dependent on young flush and thus Asian citrus psyllid production po...

  15. Proposal to conserve the name Phomopsis citri H.S. Fawc. (Diaporthe citri), with a conserved type, against Phomopsis citri (Sacc.) Traverso & Spessa (Ascomycota, Diaporthales, Diaporthaceae)

    Science.gov (United States)

    The name Diaporthe citri applies to a fungus that causes a disease on Citrus known as melanose or stem end rot of mature fruit after harvest and occurs widely in North America and Asia. Initially described as the illegitimate Phomopsis citri H.S. Fawc. 1912, non P. citri (Sacc.) Traverso & Spessa 19...

  16. Foliar application of biofilm formation-inhibiting compounds enhances control of citrus canker caused by Xanthomonas citri subsp. citri.

    Science.gov (United States)

    Li, Jinyun; Wang, Nian

    2014-02-01

    Citrus canker caused by the bacterium Xanthomonas citri subsp. citri is an economically important disease of citrus worldwide. Biofilm formation plays an important role in early infection of X. citri subsp. citri on host leaves. In this study, we assessed the hypothesis that small molecules inhibiting biofilm formation reduce X. citri subsp. citri infection and enhance the control of citrus canker disease. D-leucine and 3-indolylacetonitrile (IAN) were found to prevent biofilm formation by X. citri subsp. citri on different abiotic surfaces and host leaves at a concentration lower than the minimum inhibitory concentration (MIC). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis indicated that IAN repressed expression of chemotaxis/motility-related genes in X. citri subsp. citri. In laboratory experiments, planktonic and biofilm cells of X. citri subsp. citri treated with D-leucine and IAN, either alone or in combination, were more susceptible to copper (CuSO4) than those untreated. In greenhouse assays, D-leucine and IAN applied alone or combined with copper reduced both the number of canker lesions and bacterial populations of X. citri subsp. citri on citrus host leaves. This study provides the basis for the use of foliar-applied biofilm inhibitors for the control of citrus canker alone or combined with copper-based bactericides.

  17. Fortunella margarita Transcriptional Reprogramming Triggered by Xanthomonas citri subsp. citri

    Directory of Open Access Journals (Sweden)

    Khalaf Abeer A

    2011-11-01

    Full Text Available Abstract Background Citrus canker disease caused by the bacterial pathogen Xanthomonas citri subsp. citri (Xcc has become endemic in areas where high temperature, rain, humidity, and windy conditions provide a favourable environment for the dissemination of the bacterium. Xcc is pathogenic on many commercial citrus varieties but appears to elicit an incompatible reaction on the citrus relative Fortunella margarita Swing (kumquat, in the form of a very distinct delayed necrotic response. We have developed subtractive libraries enriched in sequences expressed in kumquat leaves during both early and late stages of the disease. The isolated differentially expressed transcripts were subsequently sequenced. Our results demonstrate how the use of microarray expression profiling can help assign roles to previously uncharacterized genes and elucidate plant pathogenesis-response related mechanisms. This can be considered to be a case study in a citrus relative where high throughput technologies were utilized to understand defence mechanisms in Fortunella and citrus at the molecular level. Results cDNAs from sequenced kumquat libraries (ESTs made from subtracted RNA populations, healthy vs. infected, were used to make this microarray. Of 2054 selected genes on a customized array, 317 were differentially expressed (P Conclusion Functional categorization of kumquat Xcc-responsive genes revealed an enhanced defence-related metabolism as well as a number of resistant response-specific genes in the kumquat transcriptome in response to Xcc inoculation. Gene expression profile(s were analyzed to assemble a comprehensive and inclusive image of the molecular interaction in the kumquat/Xcc system. This was done in order to elucidate molecular mechanisms associated with the development of the hypersensitive response phenotype in kumquat leaves. These data will be used to perform comparisons among citrus species to evaluate means to enhance the host immune responses

  18. Plant resistance within the Rutaceae to Asian citrus psyllid

    Science.gov (United States)

    USDA-ARS recently initiated research on host plant resistance to the Asian citrus psyllid. The psyllid is an important invasive pest of citrus in the United States because it transmits a serious disease of citrus known as huanglongbing (citrus greening). There is no cure for this bacterial disease. ...

  19. Restriction digestion method for haplotyping the potato psyllid, Bactericera cockerelli

    Science.gov (United States)

    A restriction digestion method has been developed for haplotyping the potato psyllid, Bactericera cockerelli Sulc., an economically important pest of solanaceous crops. This method differentiates the four known potato psyllid haplotypes by utilizing restriction enzyme digestion of a portion of the ...

  20. Behavioral assay on Asian citrus psyllid attraction to orange jasmine

    Science.gov (United States)

    The Asian citrus psyllid (ACP) is an important pest because it transmits a bacterium putatively responsible for huanglongbing, a devastating citrus disease. Research on ACP chemical ecology is of interest with respect to identifying attractants and repellents for managing the psyllid. We report on a...

  1. Screening of LED light source of the adapter solar trap lamp for trapping the citrus psyllid%适配太阳能诱虫器诱杀柑橘木虱LED光源的筛选

    Institute of Scientific and Technical Information of China (English)

    林雄杰; 范国成; 胡菡青; 阮传清; 蔡子坚; Xia Yulu; 杜云贵; 刘波

    2013-01-01

    The citrus psyllid is the vector of Huanglongbing.In order to control citrus psyllid and prevent disease,screening of LED light source adapter solar trap lamp for citrus psyllid by using reaction vessel of insect behavior was carried out in the laboratory.The results showed that the phototaxis of 1 800 1x and 4 310 lx blue and green light sources (wavelength 460 nm and 531 nm) were best on the illumination duration of 17 h.It was also found that the phototaxis of citrus psyllid adults was positively correlated with light intensity and light duration under the same wavelength condition.It might provide an effective theoretical basis for the LED light source for forecasting and trapping citrus psyllid in the field in the future.%柑橘木虱(Dia phorina citri)是柑橘黄龙病的传播媒介.为达到治虫防病的目的,在室内利用昆虫行为反应器开展适配太阳能诱虫器的LED光源筛选,结果表明:柑橘木虱成虫对光照强度分别为1 800 lx和4 310 lx的蓝光和绿光(波长分别为460 nm和531 nm)在光照时长为17 h时的趋光性最佳,相同波长条件下,其趋光性与光照强度和光照时长成正相关.该研究结果为进一步探索适宜田间柑橘木虱测报和高效诱杀的LED光源提供理论依据.

  2. Impacts of the Psyllid Arytinnis hakani (Homoptera: Psyllidae) on Invasive French Broom in Relation to Plant Size and Psyllid Density.

    Science.gov (United States)

    Hogg, Brian N; Moran, Patrick J; Smith, Lincoln

    2017-06-01

    The impacts of weed biological control agents may vary with plant ontogeny. As plants grow, structural and chemical changes can alter plant resistance, which may reduce herbivory via chemical or structural defenses, and plant tolerance, which may enable plants to maintain fitness despite attack. Resistance and tolerance generally increase as plants grow. Nonetheless, prerelease tests of agent efficacy often overlook plant ontogeny. Here, we assess the performance and impacts of a candidate biocontrol agent, the psyllid Arytinnis hakani (Loginova), in relation to the age of its host plant, the invasive shrub French broom, Genista monspessulana. We also examined whether the psyllid can consistently kill plants when its densities are sufficiently high. Survival of psyllids to adulthood and the timing of adult emergence did not differ between plant sizes, indicating that performance of nymphs was not influenced by plant size. However, adult psyllid survival was reduced on small plants, suggesting that nymphs and adults responded differently to ontogenetic changes in plant quality. Psyllids affected the growth of small and large plants similarly; all measured plant growth parameters were lower in the presence of psyllids regardless of plant size. In a separate experiment, effects on plant survival depended on psyllid density, as higher realized densities of ∼9 psyllids per cm stem length were necessary to consistently kill plants. Thus, results suggest that the psyllid would be equally effective on a range of plant sizes, particularly at high densities, and show the potential of the psyllid to help control French broom in California. Published by Oxford University Press on behalf of Entomological Society of America 2017. This work is written by US Government employees and is in the public domain in the US.

  3. 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.

  4. Complete Genome Sequences of Six Copper-Resistant Xanthomonas citri pv.?citri Strains Causing Asiatic Citrus Canker, Obtained Using Long-Read Technology

    OpenAIRE

    Richard, Damien; Boyer, Claudine; Vernière, Christian; Canteros, B.I.; Lefeuvre, Pierre; Pruvost, Olivier

    2017-01-01

    The gammaproteobacterium Xanthomonas citri pv. citri causes Asiatic citrus canker. Pathotype A strains have a broad host range, which includes most commercial citrus species, and they cause important economic losses worldwide. Control often relies on frequent copper sprays. We present here the complete genomes of six X. citri pv. citri copper-resistant strains. (Résumé d'auteur)

  5. Complete Genome Sequences of Six Copper-Resistant Xanthomonas citri pv. citri Strains Causing Asiatic Citrus Canker, Obtained Using Long-Read Technology

    Science.gov (United States)

    Richard, Damien; Boyer, Claudine; Vernière, Christian; Canteros, Blanca I.; Lefeuvre, Pierre

    2017-01-01

    ABSTRACT The gammaproteobacterium Xanthomonas citri pv. citri causes Asiatic citrus canker. Pathotype A strains have a broad host range, which includes most commercial citrus species, and they cause important economic losses worldwide. Control often relies on frequent copper sprays. We present here the complete genomes of six X. citri pv. citri copper-resistant strains. PMID:28336584

  6. The cys regulon of Xanthomonas citri

    Energy Technology Data Exchange (ETDEWEB)

    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)

  7. Gender- and species-specific characteristics of bacteriomes from three psyllid species (Hemiptera: Psylloidae)

    Science.gov (United States)

    Psyllids (Hemiptera: Pyslloidea) harbor bacterial symbionts in specialized organs called bacteriomes. Bacteriomes may be subject to manipulation to control psyllid pests including Bactericera cockerelli (Sulc) (Hemiptera: Triozidae) and Cacopsylla pyricola (Forster) (Hemiptera: Psyllidae) if the bi...

  8. Competition between honeydew producers in an ant-hemipteran interaction may enhance biological control of an invasive pest.

    Science.gov (United States)

    Tena, A; Hoddle, C D; Hoddle, M S

    2013-12-01

    Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is an invasive citrus pest in southern California, which secretes honeydew and has the potential to spread a lethal bacterial disease, huanglongbing, of citrus. In urban citrus, Argentine ant, Linepithema humile (Mayr) (Hymenoptera: Formicidae), also an invasive pest, tends honeydew-producing hemipterans. We used field data to determine whether the mutualistic relationship between L. humile and six established species of honeydew producers may hinder or favor the establishment of D. citri and its biological control with Tamarixia radiata (Waterston) (Hymenoptera: Eulophidae) in citrus via competition or mutualism for ants, respectively. In the field, L. humile and D. citri are engaged in a mutualistic relationship. Ants harvest solid honeydew secreted by psyllid nymphs and tended more than 55% of observed D. citri colonies. Linepithema humile displayed a preference hierarchy when tending honeydew producers infesting citrus. It responded equally or less intensively to D. citri than to other honeydew-producing species. Consequently, the mutualism between L. humile and D. citri was affected by the presence of other honeydew-producing species, and the percentage of D. citri colonies tended by L. humile. The number of ants per D. citri colony also decreased as the number of other honeydew producers increased. Diaphorina citri density was also affected by the presence of other honeydew producers. Both colony size and the number of D. citri nymphs counted per tree decreased as the number of other honeydew producers increased. Our results indicate that competition between honeydew producers for the mutualist ant L. humile may hinder the establishment of D. citri by possibly facilitating increased biological control.

  9. Characterization of a variant of Xanthomonas citri subsp. citri that triggers a host-specific defense response.

    Science.gov (United States)

    Chiesa, María A; Siciliano, María F; Ornella, Leonardo; Roeschlin, Roxana A; Favaro, María A; Delgado, Natalia Pino; Sendín, Lorena N; Orce, Ingrid G; Ploper, L Daniel; Vojnov, Adrian A; Vacas, José Gadea; Filippone, María P; Castagnaro, Atilio P; Marano, María R

    2013-06-01

    Citrus is an economically important fruit crop that is severely afflicted by Asiatic citrus bacterial canker (CBC), a disease caused by the phytopathogen Xanthomonas citri subsp. citri (X. citri). To gain insight into the molecular epidemiology of CBC, 42 Xanthomonas isolates were collected from a range of Citrus spp. across 17 different orchards in Tucumán, Argentina and subjected to molecular, biochemical, and pathogenicity tests. Analysis of genome-specific X. citri markers and DNA polymorphisms based on repetitive elements-based polymerase chain reaction showed that all 42 isolates belonged to X. citri. Interestingly, pathogenicity tests showed that one isolate, which shares >90% genetic similarity to the reference strain X. citri T, has host range specificity. This new variant of X. citri subsp. citri, named X. citri A(T), which is deficient in xanthan production, induces an atypical, noncankerous chlorotic phenotype in Citrus limon and C. paradisi and weak cankerous lesions in C. aurantifolia and C. clementina leaves. In C. limon, suppression of canker development is concomitant with an oxidative burst; xanthan is not implicated in the phenotype induced by this interaction, suggesting that other bacterial factors would be involved in triggering the defense response.

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

    Directory of Open Access Journals (Sweden)

    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.

  11. Entomopathogenic Fungi (Hypocreales) for Control of Potato Psyllid

    Science.gov (United States)

    Laboratory and field trials of the four isolates of entomopathogenic fungi, Isaria fumosorosea and Metarhizium anisopliae were conducted to evaluate their potential for control of the potato psyllid. In the laboratory, 2 ml aqueous suspension of 107conidia/ml in a of each isolate applied in a Potte...

  12. Potato psyllid vector of zebra chip disease in Pacific Northwest

    Science.gov (United States)

    Zebra chip is a destructive disease of potatoes in the Pacific Northwest and other potato production regions of North America. The pathogen associated with this disease is transmitted by the potato psyllid. A team of researchers which included a scientist at the ARS in Wapato, WA updated an extens...

  13. Scientific Opinion on the risk to plant health of Xanthomonas citri pv. citri and Xanthomonas citri pv. aurantifolii for the EU territory

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Plant Health (PLH

    2014-02-01

    Full Text Available The Panel conducted a pest risk assessment for Xanthomonas campestris (all strains pathogenic to Citrus for the EU territory and an assessment of the effectiveness of present EU requirements against Xanthomonas strains pathogenic to citrus. The risk assessment was conducted under the scenario of absence of the current specific EU plant health legislation and the assumption that citrus-exporting countries apply measures to reduce yield and quality losses. Risk reduction options were systematically identified and evaluated. The strains of X. campestris pathogenic to citrus have been reclassified as four distinct infraspecific taxa within two species: X. citri and X. alfalfae. Only two pathovars (X. citri pv. citri and X. citri pv. aurantifolii are responsible for the citrus bacterial canker thatpresents a major risk for the citrus industry in the EU. Seven entry pathways have been identified and evaluated. The likelihood of entry was rated unlikely for fruit, very likely for fruit plants for planting, moderately likely for ornamental plants for planting and unlikely for leaves and twigs. The uncertainty of probability of entry was rated as high. The probability of establishment was rated as moderately likely to likely with a medium uncertainty because host plants are widely present in EU areas where environmental conditions are suitable. Once established, spread would be likely with a low uncertainty. The impact of the disease, even if control measures are applied, was rated as moderate to major with a medium uncertainty. The disease would cause yield losses in areas where citrus is the main crop, increase the need for control measures and create environmental problems.The combined EU regulations have been shown to be effective in preventing the introduction of X. citri pv. citri or X. citri pv. aurantifolii in the EU, as no outbreaks of citrus canker in the EU territory have been reported.

  14. Selection of Small Synthetic Antimicrobial Peptides Inhibiting Xanthomonas citri subsp. citri Causing Citrus Canker

    Science.gov (United States)

    Choi, Jeahyuk; Park, Euiho; Lee, Se-Weon; Hyun, Jae-Wook; Baek, Kwang-Hyun

    2017-01-01

    Citrus canker disease decreases the fruit quality and yield significantly, furthermore, emerging of streptomycin-resistant pathogens threatens the citrus industry seriously because of a lack of proper control agents. Small synthetic antimicrobial peptides (AMPs) could be a promising alternative. Fourteen hexapeptides were selected by using positional scanning of synthetic peptide combinatorial libraries. Each hexapeptide showed different antimicrobial spectrum against Bacillus, Pseudomonas, Xanthomonas, and Candida species. Intriguingly, BHC10 showed bactericidal activity exclusively on Xanthomonas citri subsp. citri (Xcc), while BHC7 was none-active exclusively against two Pseudomonas spp. at concentration of 100 μg/ml suggesting potential selectivity constrained in hexapeptide frame. Three hexapeptides, BHC02, 06 and 11, showed bactericidal activities against various Xcc strains at concentration of 10 μg/ml. When they were co-infiltrated with pathogens into citrus leaves the disease progress was suppressed significantly. Further study would be needed to confirm the actual disease control capacity of the selected hexapeptides. PMID:28167892

  15. Matrimony vine and potato psyllid in the Pacific Northwest: a worrisome marriage?

    Science.gov (United States)

    Managing zebra chip disease in the potato growing regions of Washington, Oregon, and Idaho is complicated by confusion about the source of the insect vector (potato psyllid) as it colonizes potato fields in these growing regions. Not knowing where the psyllid is before arriving in Washington potato...

  16. Field survey of Asian citrus psyllid (Hemiptera: Liviidae) infestations associated with six cultivars of Poncirus trifoliata

    Science.gov (United States)

    The Asian citrus psyllid is an important pest because it transmits a bacterium responsible for huanglongbing, also known as citrus greening disease. Insecticidal control of the psyllid is a key tactic used to manage the disease, but host plant resistance may hold some promise as an alternative. Resu...

  17. Coccidoxenoides perminutus parasitizing Planococcus citri on vine in Brazil

    Directory of Open Access Journals (Sweden)

    Maria Herlândia de Araújo Fernandes

    2016-07-01

    Full Text Available ABSTRACT: From those of the mealybug family, the citrus mealybug, Planococcus citriRisso, 1813 (Hemiptera: Pseudococcidae is frequently held responsible for various diseases occurring in vineyards. This is a first time report in Brazil, on the occurrence of a parasitoid controlling P. citri. In our search for the natural enemies of P. citri, bunches of grapes with the vine were collected every week for one month, in those regions where table grapes were produced, like Petrolina, Pernambuco, Brazil. In these samples, among the natural pest infestations Coccidoxenoides perminutusGirault, 1915 (Hymenoptera: Encyrtidae was the only natural enemy species reported. In this research, we attempted to study the potential of parasitism as well as the strategies that this natural agent uses in pest control.

  18. 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/.

  19. Unusual Behaviour — Unusual Morphology: Mutualistic Relationships between Ants (Hymenoptera: Formicidae) and Diaphorina enderleini (Hemiptera: Psylloidea), Associated with Vernonia amygdalina (Asteraceae)

    National Research Council Canada - National Science Library

    Désirée Chantal Aléné; Champlain Djiéto-Lordon; Daniel Burckhardt

    2011-01-01

    .... The lack of wax deprives them of the protection common in various free-living psyllids. Consequently the psyllid is involved in an unusual mutualistic interaction with the ants Pheidole megacephala and Crematogaster striatula...

  20. treA Codifies for a Trehalase with Involvement in Xanthomonas citri subsp. citri Pathogenicity

    Science.gov (United States)

    Alexandrino, André Vessoni; Goto, Leandro Seiji; Novo-Mansur, Maria Teresa Marques

    2016-01-01

    Citrus canker, caused by the bacterium Xanthomonas citri subsp. citri (Xcc), is a severe disease of citrus. Xcc presents broad spectrum of citrus hosts including economically important species whereas X. fuscans subsp. aurantifolii–type C (XauC) causes a milder disease and only infects Citrus aurantifolia. Trehalase catalyzes hydrolysis of the disaccharide trehalose, a sugar that has been reported to be related to Xcc pathogenicity. We expressed the recombinant gene product and assessed Xcc trehalase structural and kinetics data. The recombinant protein presented 42.7% of secondary structures in α-helix and 13% in β-sheets, no quaternary structure in solution, and Michaelis-Menten constant (KM) of 0.077 mM and Vmax 55.308 μMol glucose.min-1.mg protein-1 for trehalose. A Xcc mutant strain (XccΔtreA) was produced by gene deletion from Xcc genome. Enzymatic activity of trehalase was determined in Xcc, XauC and XccΔtreA cellular lysates, showing the highest values for XauC in in vitro infective condition and no activity for XccΔtreA. Finally, leaves of Citrus aurantifolia infected with XccΔtreA showed much more drenching and necrosis than those infected by wild type Xcc. We concluded that trehalase contributes to alleviate bacterial virulence and that inability for trehalose hydrolysis may promote higher Xcc infectivity. PMID:27611974

  1. treA Codifies for a Trehalase with Involvement in Xanthomonas citri subsp. citri Pathogenicity.

    Science.gov (United States)

    Alexandrino, André Vessoni; Goto, Leandro Seiji; Novo-Mansur, Maria Teresa Marques

    2016-01-01

    Citrus canker, caused by the bacterium Xanthomonas citri subsp. citri (Xcc), is a severe disease of citrus. Xcc presents broad spectrum of citrus hosts including economically important species whereas X. fuscans subsp. aurantifolii-type C (XauC) causes a milder disease and only infects Citrus aurantifolia. Trehalase catalyzes hydrolysis of the disaccharide trehalose, a sugar that has been reported to be related to Xcc pathogenicity. We expressed the recombinant gene product and assessed Xcc trehalase structural and kinetics data. The recombinant protein presented 42.7% of secondary structures in α-helix and 13% in β-sheets, no quaternary structure in solution, and Michaelis-Menten constant (KM) of 0.077 mM and Vmax 55.308 μMol glucose.min-1.mg protein-1 for trehalose. A Xcc mutant strain (XccΔtreA) was produced by gene deletion from Xcc genome. Enzymatic activity of trehalase was determined in Xcc, XauC and XccΔtreA cellular lysates, showing the highest values for XauC in in vitro infective condition and no activity for XccΔtreA. Finally, leaves of Citrus aurantifolia infected with XccΔtreA showed much more drenching and necrosis than those infected by wild type Xcc. We concluded that trehalase contributes to alleviate bacterial virulence and that inability for trehalose hydrolysis may promote higher Xcc infectivity.

  2. A protein expression system for tandem affinity purification in Xanthomonas citri subsp. citri

    Directory of Open Access Journals (Sweden)

    Giordanni C. Dantas

    2016-06-01

    Full Text Available Abstract Citrus canker, caused by the Gram-negative bacterium Xanthomonas citri subsp. citri (Xac, is one of the most devastating diseases to affect citrus crops. There is no treatment for citrus canker; effective control against the spread of Xac is usually achieved by the elimination of affected plants along with that of asymptomatic neighbors. An in depth understanding of the pathogen is the keystone for understanding of the disease; to this effect we are committed to the development of strategies to ease the study of Xac. Genome sequencing and annotation of Xac revealed that ∼37% of the genome is composed of hypothetical ORFs. To start a systematic characterization of novel factors encoded by Xac, we constructed integrative-vectors for protein expression specific to this bacterium. The vectors allow for the production of TAP-tagged proteins in Xac under the regulation of the xylose promoter. In this study, we show that a TAP-expression vector, integrated into the amy locus of Xac, does not compromise its virulence. Furthermore, our results also demonstrate that the polypeptide TAP can be overproduced in Xac and purified from the soluble phase of cell extracts. Our results substantiate the use of our vectors for protein expression in Xac thus contributing a novel tool for the characterization of proteins and protein complexes generated by this bacterium in vivo.

  3. Bioactive Organocopper Compound from Pseudomonas aeruginosa Inhibits the Growth of Xanthomonas citri subsp. citri

    Science.gov (United States)

    de Oliveira, Admilton G.; Spago, Flavia R.; Simionato, Ane S.; Navarro, Miguel O. P.; da Silva, Caroline S.; Barazetti, André R.; Cely, Martha V. T.; Tischer, Cesar A.; San Martin, Juca A. B.; de Jesus Andrade, Célia G. T.; Novello, Cláudio R.; Mello, João C. P.; Andrade, Galdino

    2016-01-01

    Citrus canker is a very destructive disease of citrus species. The challenge is to find new compounds that show strong antibiotic activity and low toxicity to plants and the environment. The objectives of the present study were (1) to extract, purify and evaluate the secondary metabolites with antibiotic activity produced by Pseudomonas aeruginosa LV strain in vitro against Xanthomonas citri subsp. citri (strain 306), (2) to determine the potential of semi-purified secondary metabolites in foliar application to control citrus canker under greenhouse conditions, and (3) to identify antibiotic activity in orange leaf mesophyll infected with strain 306, by electron microscopy. Two pure bioactive compounds were isolated, an organocopper antibiotic compound (OAC) and phenazine-1-carboxamide. Phenazine-1-carboxamide did not show any antibiotic activity under the experimental conditions used in this study. The OAC showed a high level of antibiotic activity with a minimum inhibitory concentration of 0.12 μg mL-1. In greenhouse tests for control of citrus canker in orange trees, the semi-purified fraction F3d reduced lesion formation by about 97%. The concentration used was 500 times lower than that for the recommended commercial copper-based product. Electron microscopy showed that F3d altered the exopolysaccharide matrix and caused cell lysis of the pathogen inside the citrus canker lesions. These results suggest that secondary metabolites produced by inducing P. aeruginosa LV strain have a high potential to be used as a bioproduct to control citrus canker. PMID:26903992

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

    Directory of Open Access Journals (Sweden)

    Vahideh Hasabi

    2014-06-01

    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.

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

    Directory of Open Access Journals (Sweden)

    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

  6. 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

  7. The Effect of Citrus Essential Oils and Their Constituents on Growth of Xanthomonas citri subsp. citri.

    Science.gov (United States)

    Mirzaei-Najafgholi, Hossein; Tarighi, Saeed; Golmohammadi, Morteza; Taheri, Parissa

    2017-04-14

    Citrus bacterial canker (CBC) caused by Xanthomonas citri subsp. citri (Xcc), is the most devastating of the citrus diseases worldwide. During our study, we found that Essential oils (EOs) of some citrus cultivars are effective on Xcc. Therefore, it prompted us to determine the plant metabolites responsible for the antibacterial properties. We obtained EOs from some locally cultivated citrus by using a Clevenger apparatus and their major constituents were identified by gas chromatography/mass spectrometry (GC-MS). The effect of Citrus aurantium, C. aurantifolia, Fortunella sp. EOs and their major constituents were evaluated against Xcc-KVXCC1 using a disk diffusion assay. Minimal inhibitory and bactericidal concentration of the EOs and their constituents were determined using the broth microdilution method. C. aurantium, C. aurantifolia Eos, and their major constituents including citral, linalool, citronellal, geraniol, α-terpineol, and linalyl acetate indicated antibacterial effects against Xcc. The C. aurantifolia EO and citral showed the highest antibacterial activity among the tested EOs and constituents with inhibition zones of 15 ± 0.33 mm and 16.67 ± 0.88 mm, respectively. Synergistic effects of the constituents were observed between α-terpineol-citral, citral-citronellal, citral-geraniol, and citronellal-geraniol by using a microdilution checkerboard assay. Transmission electron microscopy revealed that exposure of Xcc cells to citral caused cell wall damage and altered cytoplasmic density. We introduced C. aurantifolia and C. aurantium EOs, and their constituents citral, α-terpineol, citronellal, geraniol, and linalool as possible control agents for CBC.

  8. Proteome of the phytopathogen Xanthomonas citri subsp. citri: a global expression profile

    Directory of Open Access Journals (Sweden)

    Ferro Jesus A

    2010-11-01

    Full Text Available Abstract Background Citrus canker is a disease caused by Xantomonas citri subsp.citri (Xac, and has emerged as one of the major threats to the worldwide citrus crop because it affects all commercial citrus varieties, decreases the production and quality of the fruits and can spread rapidly in citrus growing areas. In this work, the first proteome of Xac was analyzed using two methodologies, two-dimensional liquid chromatography (2D LC and tandem mass spectrometry (MS/MS. Results In order to gain insight into the metabolism of Xac, cells were grown on two different media (NB - Nutrient Broth and TSE - Tryptone Sucrose broth enriched with glutamic acid, and proteins were proteolyzed with trypsin and examined by 2D LC-MS/MS. Approximately 39% of all predicted proteins by annotation of Xac were identified with their component peptides unambiguously assigned to tandem mass spectra. The proteins, about 1,100, were distributed in all annotated functional categories. Conclusions This is the first proteomic reference map for the most aggressive strain of Xanthomonas pathogen of all orange varieties. The compilation of metabolic pathways involved with bacterial growth showed that Xac expresses a complete central and intermediary metabolism, replication, transcription and translation machineries and regulation factors, distinct membrane transporters (ABC, MFS and pumps and receptors (MCP, TonB dependent and metabolites acquisition, two-component systems (sensor and regulatory components and response regulators. These data corroborate the growth curve in vitro and are the first reports indicating that many of these genome annotated genes are translated into operative in Xac. This proteomic analysis also provided information regarding the influence of culture medium on growth and protein expression of Xac.

  9. Non-host defense response in a novel Arabidopsis-Xanthomonas citri subsp. citri pathosystem.

    Directory of Open Access Journals (Sweden)

    Chuanfu An

    Full Text Available Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc, is one of the most destructive diseases of citrus. Progress of breeding citrus canker-resistant varieties is modest due to limited resistant germplasm resources and lack of candidate genes for genetic manipulation. The objective of this study is to establish a novel heterologous pathosystem between Xcc and the well-established model plant Arabidopsis thaliana for defense mechanism dissection and resistance gene identification. Our results indicate that Xcc bacteria neither grow nor decline in Arabidopsis, but induce multiple defense responses including callose deposition, reactive oxygen species and salicylic aicd (SA production, and defense gene expression, indicating that Xcc activates non-host resistance in Arabidopsis. Moreover, Xcc-induced defense gene expression is suppressed or attenuated in several well-characterized SA signaling mutants including eds1, pad4, eds5, sid2, and npr1. Interestingly, resistance to Xcc is compromised only in eds1, pad4, and eds5, but not in sid2 and npr1. However, combining sid2 and npr1 in the sid2npr1 double mutant compromises resistance to Xcc, suggesting genetic interactions likely exist between SID2 and NPR1 in the non-host resistance against Xcc in Arabidopsis. These results demonstrate that the SA signaling pathway plays a critical role in regulating non-host defense against Xcc in Arabidopsis and suggest that the SA signaling pathway genes may hold great potential for breeding citrus canker-resistant varieties through modern gene transfer technology.

  10. Amplified fragment length polymorphism and multilocus sequence analysis-based genotypic relatedness among pathogenic variants of Xanthomonas citri pv. citri and Xanthomonas campestris pv. bilvae.

    Science.gov (United States)

    Bui Thi Ngoc, Lan; Vernière, Christian; Jouen, Emmanuel; Ah-You, Nathalie; Lefeuvre, Pierre; Chiroleu, Frédéric; Gagnevin, Lionel; Pruvost, Olivier

    2010-03-01

    Three pathogenic variants (i.e. pathotypes) have been described within Xanthomonas citri pv. citri, the causal agent of Asiatic citrus canker. Pathotype A strains naturally infect a wide range of Citrus species and members of some related genera. In contrast, pathotypes A* and A(w) have narrow host ranges within the genus Citrus and have been isolated from Mexican lime (Citrus aurantifolia L.) and from Mexican lime and alemow (Citrus macrophylla L.), respectively. We used amplified fragment length polymorphism (AFLP) and multilocus sequence analysis (MLSA) based on four partial housekeeping gene sequences (atpD, dnaK, efp and gyrB ) for the genotypic classification of Xanthomonas citri pv. citri and the poorly characterized citrus pathogen Xanthomonas campestris pv. bilvae. A Mantel test showed that genetic distances derived from AFLP and MLSA were highly correlated. X. campestris pv. bilvae showed a close relatedness to the type strain of X. citri, indicating that this pathovar should be reclassified as X. citri pv. bilvae. All pathotype A* and A(w) strains were most closely related to X. citri pv. citri strains with a wide host range (pathotype A), confirming previous DNA-DNA hybridization data. Pathotype A(w) should be considered a junior synonym of pathotype A* on the basis of pathogenicity tests, AFLP, MLSA and PCR using pathovar-specific primers. Evolutionary genome divergences computed from AFLP data suggested that pathotype A* (including A(w) strains) is a group of strains that shows a wider genetic diversity than pathotype A.

  11. Asymptomatic spread of huanglongbing and implications for disease control.

    Science.gov (United States)

    Lee, Jo Ann; Halbert, Susan E; Dawson, William O; Robertson, Cecile J; Keesling, James E; Singer, Burton H

    2015-06-16

    Huanglongbing (HLB) is a bacterial infection of citrus trees transmitted by the Asian citrus psyllid Diaphorina citri. Mitigation of HLB has focused on spraying of insecticides to reduce the psyllid population and removal of trees when they first show symptoms of the disease. These interventions have been only marginally effective, because symptoms of HLB do not appear on leaves for months to years after initial infection. Limited knowledge about disease spread during the asymptomatic phase is exemplified by the heretofore unknown length of time from initial infection of newly developing cluster of young leaves, called flush, by adult psyllids until the flush become infectious. We present experimental evidence showing that young flush become infectious within 15 d after receiving an inoculum of Candidatus Liberibacter asiaticus (bacteria). Using this critical fact, we specify a microsimulation model of asymptomatic disease spread and intensity in a grove of citrus trees. We apply a range of psyllid introduction scenarios to show that entire groves can become infected with up to 12,000 psyllids per tree in less than 1 y, before most of the trees show any symptoms. We also show that intervention strategies that reduce the psyllid population by 75% during the flushing periods can delay infection of a full grove, and thereby reduce the amount of insecticide used throughout a year. This result implies that psyllid surveillance and control, using a variety of recently available technologies, should be used from the initial detection of invasion and throughout the asymptomatic period.

  12. The small and large subunits of carbamoyl-phosphate synthase exhibit diverse contributions to pathogenicity in Xanthomonas citri subsp. citri

    Institute of Scientific and Technical Information of China (English)

    Guo Jing; SonG Xue; Zou Li-fang; Zou Hua-song; CHen Gong-you

    2015-01-01

    Carbamoyl-phosphate synthase plays a vital role in the carbon and nitrogen metabolism cycles. In Xanthomonas citri subsp. citri, carA and carB encode the smal and large subunits of carbamoyl-phosphate synthase, respectively. The deletion mutation of the coding regions revealed that carA did not affect any of the phenotypes, while carB played multiple roles in pathogenicity. The deletion of carB rendered the loss of pathogenicity in host plants and the ability to induce a hyper-sensitive reaction in the non-hosts. Quantitative reverse transcription-PCR assays indicated that 11 hrp genes coding the type III secretion system were suppressed when interacting with citrus plants. The mutation in carB also affected bacterial utilization of several carbon and nitrogen resources in minimal medium MMX and extracel ular enzyme activities. These data demonstrated that only the large subunit of carbamoyl-phosphate synthase was essential for canker development by X. citri subsp. citri.

  13. Comparative ultrastructure of nonwounded Mexican lime and Yuzu leaves infected with the citrus canker bacterium Xanthomonas citri pv. citri.

    Science.gov (United States)

    Lee, In Jung; Kim, Ki Woo; Hyun, Jae Wook; Lee, Yong Hoon; Park, Eun Woo

    2009-07-01

    Ultrastructural aspects of citrus canker development were investigated in nonwounded leaves of citrus species by transmission electron microscopy (TEM). A susceptible species Mexican lime and a resistant species Yuzu were spray-inoculated with a virulent strain of Xanthomonas citri pv. citri. Initial symptoms occurred on Mexican lime approximately 9 days after inoculation, whereas they appeared on Yuzu mostly 11 days after inoculation. In Mexican lime leaves, the bacterial invasion was usually accompanied by host cell wall dissolution and cellular disruption. Fibrillar materials from degenerated cell walls were usually found in intercellular spaces. Damaged host cells with necrotic cytoplasm showed the localized separation of plasma membrane from the cell wall. Bacterial multiplication and electron-transparent capsule-like structures around bacteria were commonly observed. Meanwhile, cell wall protuberances were prominent outside host cell walls in response to bacterial invasion in Yuzu leaves. Occlusion of intercellular spaces was also formed by the fusion of two or more individual cell wall protuberances originated from two adjacent host cells. Papillae-like materials accumulated locally within host cells in close proximity to bacteria. Some bacteria were found to be undergoing degeneration in xylem vessels. Also, the shrunken, inactive bacteria were surrounded by electron-translucent fibrillar materials in intercellular spaces, implying bacterial immobilization. These cellular responses are thought to be the consequences of defense responses of Yuzu leaves to invading bacteria. In both citrus species, X. citri pv. citri contained polyphosphate bodies showing electron-dense and elliptical structures in cytoplasm.

  14. Elucidating genetic variation in Candidatus Liberibacter asiaticus transmission between Asian citrus psyllid isofemale lines

    OpenAIRE

    Dixon, Danielle; Kruse, Annie; Ramsey, John; Hosmani, Prashant; Hall, David; Saha, Surya; Ammar, El-Desouky; Cilia, Michelle; Mueller, Lukas A

    2017-01-01

    Citrus greening disease or Huanglongbing (HLB) is an epidemic bacterial infection that has devastated citrus industries across the globe. HLB leads to a reduction in fruit quality, decrease in overall fruit production and eventual death of the citrus tree.The disease is associated with this bacterium Candidatus Liberibacter asiaticus (CLas), is transmitted by the Asian citrus psyllid (ACP).Objective 1: Improve the current psyllid genome through manual curation.Objective 2: Characterize genet...

  15. Unravelling potential virulence factor candidates in Xanthomonas citri. subsp. citri by secretome analysis

    Directory of Open Access Journals (Sweden)

    Rafael M. Ferreira

    2016-02-01

    Full Text Available Citrus canker is a major disease affecting citrus production in Brazil. It’s mainly caused by Xanthomonas citri subsp. citri strain 306 pathotype A (Xac. We analysed the differential expression of proteins secreted by wild type Xac and an asymptomatic mutant for hrpB4 (ΔhrpB4 grown in Nutrient Broth (NB and a medium mimicking growth conditions in the plant (XAM1. This allowed the identification of 55 secreted proteins, of which 37 were secreted by both strains when cultured in XAM1. In this secreted protein repertoire, the following stand out: Virk, Polyphosphate-selective porin, Cellulase, Endoglucanase, Histone-like protein, Ribosomal proteins, five hypothetical proteins expressed only in the wild type strain, Lytic murein transglycosylase, Lipoprotein, Leucyl-tRNA synthetase, Co-chaperonin, Toluene tolerance, C-type cytochrome biogenesis membrane protein, Aminopeptidase and two hypothetical proteins expressed only in the ΔhrpB4 mutant. Furthermore, Peptidoglycan-associated outer membrane protein, Regulator of pathogenicity factor, Outer membrane proteins, Endopolygalacturonase, Chorismate mutase, Peptidyl-prolyl cis-trans isomerase and seven hypothetical proteins were detected in both strains, suggesting that there was no relationship with the secretion mediated by the type III secretory system, which is not functional in the mutant strain. Also worth mentioning is the Elongation factor Tu (EF-Tu, expressed only the wild type strain, and Type IV pilus assembly protein, Flagellin (FliC and Flagellar hook-associated protein, identified in the wild-type strain secretome when grown only in NB. Noteworthy, that FliC, EF-Tu are classically characterized as PAMPs (Pathogen-associated molecular patterns, responsible for a PAMP-triggered immunity response. Therefore, our results highlight proteins potentially involved with the virulence. Overall, we conclude that the use of secretome data is a valuable approach that may bring more knowledge of the

  16. Improved real-time PCR diagnosis of citrus stubborn disease by targeting prophage genes of Spiroplasma citri

    Science.gov (United States)

    Spiroplasma citri is a phloem-limited bacterium causing citrus stubborn disease (CSD). Isolation and culturing of S. citri is difficult and time consuming. Current detection methods use polymerase chain reaction (PCR) assays with primers developed from sequences of S. citri house-keeping genes. In c...

  17. First report of citrus canker caused by Xanthomonas citri in Somalia

    Science.gov (United States)

    Xanthomonas citri, causal agent of citrus canker, has been reported in several countries in Africa, but not Somalia. During 2006 and 2007, hyperplasia-type lesions, often surrounded by a water-soaked margin and yellow halo, typical of citrus canker caused by X. citri, were found on 8-10 year-old gr...

  18. Comparison of Droplet Digital PCR and Quantitative PCR Assays for Quantitative Detection of Xanthomonas citri Subsp. citri.

    Directory of Open Access Journals (Sweden)

    Yun Zhao

    Full Text Available Droplet digital polymerase chain reaction (ddPCR is a novel molecular biology technique providing absolute quantification of target nucleic acids without the need for an external calibrator. Despite its emerging applications in medical diagnosis, there are few reports of its use for the detection of plant pathogens. This work was designed to assess the diagnosis potential of the ddPCR for absolute quantitative detection of Xanthomonas citri subsp. citri, a quarantine plant pathogenic bacterium that causes citrus bacterial canker in susceptible Citrus species. We transferred an established quantitative PCR (qPCR assay for citrus bacterial canker diagnosis directly to the ddPCR format and compared the performance of the two methods. The qPCR assay has a broader dynamic range compared to the ddPCR assay and the ddPCR assay has a significantly higher degree of sensitivity compared to the qPCR assay. The influence of PCR inhibitors can be reduced considerably in the ddPCR assay because the collection of end-point fluorescent signals and the counting of binomial events (positive or negative droplets are associated with a Poisson algorithm. The ddPCR assay also shows lower coefficient of variation compared to the qPCR assay especially in low target concentration. The linear association of the measurements by ddPCR and qPCR assays is strong (Pearson correlation = 0.8633; P<0.001. Receiver operating characteristic analysis indicates the ddPCR methodology is a more robust approach for diagnosis of citrus bacterial canker. In summary, the results demonstrated that the ddPCR assay has the potential for the quantitative detection of X. citri subsp. citri with high precision and accuracy as compared with the results from qPCR assay. Further studies are required to evaluate and validate the value of ddPCR technology in the diagnosis of plant disease and quarantine applications.

  19. 75 FR 28233 - Availability of an Environmental Assessment for a Biological Control Agent for Asian Citrus Psyllid

    Science.gov (United States)

    2010-05-20

    ... for use as a biological control agent to reduce the severity of Asian citrus psyllid infestations. We... continental United States for use as a biological control agent to reduce the severity of Asian citrus psyllid... include chemical control and the release of an alternative biological control agent, an encyrtid...

  20. Insecticidal Activity of Entomopathogenic Fungi (Hypocreales) for Potato Psyllid, Bactericera cockerelli (Hemiptera: Triozidae): Development of Bioassay Techniques, Effect of Fungal Species and Stage of the Psyllid

    Science.gov (United States)

    The potato psyllid, Bactericera cockerelli (Šulc), is a pest of potato, tomato, and some other solanaceous vegetables and has also been incriminated in the transmission of a bacterial pathogen, Candidatus Liberibacter solanacearum, resulting in a serious disease known as “zebra chip”. Although there...

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

    Science.gov (United States)

    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.

  2. Glycaspis brimblecombei (Hem.: Psyllidae, the invasive red gum lerp psyllid recorded in Italy

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    Garonna AP

    2011-05-01

    Full Text Available During 2010 in central and southern Italy (Campania, Basilicata and Latium, the invasive red gum lerp psyllid, Glycaspis brimblecombei Moore (Hemiptera: Psyllidae, has been recorder for the first time in Italy. The psyllid was detected on Eucalyptus camaldulensis in several sampling sites. The host plant, like other species of the genus Eucalyptus, is very common in Italy as ornamental and forest species. The native country of G. brimblecombei is Australia, from which the psyllid spread throughout the world in the last decade. In the Palaearctic Region the species was first collected from the Iberian Peninsula in 2007. In this note we report some observations on the biology and distribution of this species in the new colonized Italian areas. Generalist predators, such as wasps and pirate bugs, were observed attacking G. brimblecombei, however no parasitoid activity has been detected.

  3. Comparative genomics of 43 strains of Xanthomonas citri pv. citri reveals the evolutionary events giving rise to pathotypes with different host ranges.

    Science.gov (United States)

    Gordon, Jonathan L; Lefeuvre, Pierre; Escalon, Aline; Barbe, Valérie; Cruveiller, Stéphane; Gagnevin, Lionel; Pruvost, Olivier

    2015-12-23

    The identification of factors involved in the host range definition and evolution is a pivotal challenge in the goal to predict and prevent the emergence of plant bacterial disease. To trace the evolution and find molecular differences between three pathotypes of Xanthomonas citri pv. citri that may explain their distinctive host ranges, 42 strains of X. citri pv. citri and one outgroup strain, Xanthomonas citri pv. bilvae were sequenced and compared. The strains from each pathotype form monophyletic clades, with a short branch shared by the A(w) and A pathotypes. Pathotype-specific recombination was detected in seven regions of the alignment. Using Ancestral Character Estimation, 426 SNPs were mapped to the four branches at the base of the A, A*, A(w) and A/A(w) clades. Several genes containing pathotype-specific nonsynonymous mutations have functions related to pathogenicity. The A pathotype is enriched for SNP-containing genes involved in defense mechanisms, while A* is significantly depleted for genes that are involved in transcription. The pathotypes differ by four gene islands that largely coincide with regions of recombination and include genes with a role in virulence. Both A* and A(w) are missing genes involved in defense mechanisms. In contrast to a recent study, we find that there are an extremely small number of pathotype-specific gene presences and absences. The three pathotypes of X. citri pv. citri that differ in their host ranges largely show genomic differences related to recombination, horizontal gene transfer and single nucleotide polymorphism. We detail the phylogenetic relationship of the pathotypes and provide a set of candidate genes involved in pathotype-specific evolutionary events that could explain to the differences in host range and pathogenicity between them.

  4. Acaricidal Activity of Boenninghausenia sessilicarpa Against Panonychus citri

    Institute of Scientific and Technical Information of China (English)

    YANG Hui-zhi; HU Jun-hua; LI Qing; LI Hong-jun; LIU Hao-qiang; YAO Ting-shan; RAN Chun; LEI Hui-de

    2009-01-01

    Acaricidal activity of Boenninghausenia sessilicarpa against Panonychus citri was tested in the laboratory.Four solvents were used to prepare crude extracts,petroleum ether,chloroform,ethyl acetate,and ethanol,among which ethanol was the most effective one.The results suggested that ethanol extracts of B.sessilicarpa had eminent acaricidal and ovicidal activities.Concentrated extracts were prepared using petroleum ether,chloroform,ethyl acetate,or distilled water as solvent.Mite mortality rates in the concentrated extracts by petroleum ether,chloroform,or distilled water were significantly lower than those by ethyl acetate.The LC50 values of eggs and female mites were 0.7639 and 1.1033 mg mL-1,respectively.After liquid chromatography and thin layer chromatography,the concentrated extracts were separated into 14 groups of fractions and further tests for their acaricidal and ovicidal activities were conducted.Fraction 2 was found to possess higher acaricidal and ovicidal activities.The mortality of eggs and adult mites were 85.83 and 63.07%,respectively.Moreover,fraction 2 showed moderate oviposition inhibition effect (0.8795) against P.citri when the used dose Was higher than 2.5 mg mL-1.

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

    Directory of Open Access Journals (Sweden)

    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.

  6. Effect of X-irradiation on Citrus Canker Pathogen Xanthomonas citri subsp. citri of Satsuma Mandarin Fruits

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    Min-A Song

    2015-12-01

    Full Text Available Citrus canker caused by Xanthomonas citri subsp. citri (Xcc is one of the most important bacterial diseases of citrus. Because citrus canker is not found in many countries including European Union and Australia, Xcc is strictly regulated in order to prevent its spread. In this study, the effects of X-irradiation on Xcc growth either in the suspension or on the surface of citrus fruits were investigated. The suspension containing 1×10⁷ cfu/ml of Xcc was irradiated with different absorbed doses of X-irradiation ranging from 50 to 400 Gy. The results showed that Xcc was fully dead at 400 Gy of X-irradiation. To determine the effect of X-irradiation on quarantine, the Xcc-inoculated citrus fruits were irradiated with different X-ray doses at which Xcc was completely inhibited by an irradiation dose of 250 Gy. The D₁₀ value for Xcc on citrus fruits was found to be 97 Gy, indicating the possibility of direct application on citrus quarantine without any side sterilizer. Beside, presence of Xcc on the surface of asymptomatic citrus fruits obtained from citrus canker-infected orchards was noted. It indicated that the exporting citrus fruits need any treatment so that Xcc on the citrus fruits should be completely eliminated. Based on these results, ionizing radiation can be considered as an alternative method of eradicating Xcc for export of citrus fruits.

  7. Closely-related Xanthomonas citri subsp. citri isolates trigger distinct histological and transcriptional responses in Citrus limon

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    Ingrid Georgina Orce

    Full Text Available ABSTRACT Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc, has an important economic impact on the citrus industry. Extensive information is available about the disease but, nevertheless, the study of plant-pathogen interactions could provide new information in the understanding of citrus canker disease. A new isolate has been identified, Xcc AT, which has a high genetic similarity (> 90 % to the virulent Xcc T strain based on genetic clustering analyses of the rep-PCR fingerprinting patterns, but it does not produce cankerous lesions in Citrus limon. In this study, we compared C. limon responses to Xcc AT and to the virulent Xcc T strain at both histological and transcriptional levels. Histologically, leaves inoculated with Xcc AT exhibited neither a typical disordering of the spongy mesophyll, nor a swelling of epidermis. A particular content (undetermined was also found in mesophyll cells near the stomata, together with increased starch accumulation. The transcriptomic profiles were compared by cDNA-AFLP technique. A total of 121 fragments derived from transcript (TDF were either specifically induced or repressed by the isolates, and 62 were sequenced. Analysis of global expression identified different classes of genes known to be involved in plant-pathogen interactions. This study constitutes the first approach of the specific interaction between the avirulent Xcc AT isolate and C. limon.

  8. The role as inoculum sources of Xanthomonas citri pv. citri surviving on the infected Satsuma mandarin fruits.

    Science.gov (United States)

    Kang, So Young; Kim, Ki Deok; Hong, Jeum Kyu; Hyun, He Nam; Jeun, Yong Chull

    2014-05-01

    Importing citrus fruits infected by Asiatic citrus canker caused by Xanthomonas citri pv. citri (Xcc) can act as an inoculum source for the disease epidemic in citrus canker-free countries. In this study, the pathogenicity of the causal agent of Asiatic citrus canker surviving on infected Satsuma mandarin fruits was evaluated. The washing solution of infected Satsuma mandarin fruits did not cause lesion formation on the citrus leaves. However, a typical citrus canker lesion was formed on the leaves after inoculation with higher concentrations of the inoculum from the washing solution (washing solution II). It indicated that the pathogenicity of the citrus canker surviving on the symptomatic Satsuma mandarin fruits was not changed. Scanning electron microscopic observation showed that the numbers of bacterial cells on the leaves of Satsuma mandarin which inoculated with the washing solution directly (washing solution I) was less compared to those of leaves inoculated with the washing solution II. This result spports that the pathogenicity of Xcc surviving on Satsuma mandarin fruits may not be changed but that the sucessful infection of citrus caker may depend on the concentration of the inoculum.

  9. First report of 'Candidatus Liberibacter solanacearum' associated with psyllid-affected carrots in Norway

    Science.gov (United States)

    Carrot (Daucus carota) plants with symptoms resembling those associated with the carrot psyllid Trioza apicalis and the bacterium “Candidatus Liberibacter solanacearum” were observed in 70-80% of commercial fields and experimental plots in southeastern Norway from late July to mid-September 2011; al...

  10. First report of 'Candidatus Liberibacter solanacearum' associated with psyllid-affected carrots in Sweden

    Science.gov (United States)

    Carrot (Daucus carota) plants with symptoms resembling those of the carrot psyllid Trioza apicalis and “Candidatus Liberibacter solanacearum” damage were observed in 70% of commercial fields in southern Sweden in August 2011; all cultivars grown were affected, at about 1 to 45% symptomatic plants pe...

  11. Current statewide updates regarding the battle of the Asian Citrus Psyllid and Huanglongbing

    Science.gov (United States)

    Huanglongbing (HLB) is a serious economic disease of citrus, vectored by the Asian Citrus Psyllid (ACP). The disease and its vector have become established in the United States in the last few decades. This submission reviews recent developments for pest control industry professionals. The submissio...

  12. Bindweed Psyllid: Biology, Natural History, and Interactions with the Zebra Chip Pathogen

    Science.gov (United States)

    Bactericera maculipennis is a native psyllid that commonly occurs on field bindweed in the western United States. We have found that Pacific Northwest populations of B. maculipennis carry Liberibacter solanacearum, the pathogen associated with zebra chip disease of potato. In North America, this p...

  13. Use of recombinant tobacco mosaic virus to achieve RNA interference in plants against the citrus mealybug, Planococcus citri (Hemiptera: Pseudococcidae.

    Directory of Open Access Journals (Sweden)

    Arif Muhammad Khan

    Full Text Available The citrus mealybug, Planococcus citri, is an important plant pest with a very broad plant host range. P. citri is a phloem feeder and loss of plant vigor and stunting are characteristic symptoms induced on a range of host plants, but P. citri also reduces fruit quality and causes fruit drop leading to significant yield reductions. Better strategies for managing this pest are greatly needed. RNA interference (RNAi is an emerging tool for functional genomics studies and is being investigated as a practical tool for highly targeted insect control. Here we investigated whether RNAi effects can be induced in P. citri and whether candidate mRNAs could be identified as possible targets for RNAi-based P. citri control. RNAi effects were induced in P. citri, as demonstrated by specific target reductions of P. citri actin, chitin synthase 1 and V-ATPase mRNAs after injection of the corresponding specific double-stranded RNA inducers. We also used recombinant Tobacco mosaic virus (TMV to express these RNAi effectors in Nicotiana benthamiana plants. We found that P. citri showed lower fecundity and pronounced death of crawlers after feeding on recombinant TMV-infected plants. Taken together, our data show that actin, chitin synthase 1 and V-ATPase mRNAs are potential targets for RNAi against P. citri, and that recombinant TMV is an effective tool for evaluating candidate RNAi effectors in plants.

  14. Use of recombinant tobacco mosaic virus to achieve RNA interference in plants against the citrus mealybug, Planococcus citri (Hemiptera: Pseudococcidae).

    Science.gov (United States)

    Khan, Arif Muhammad; Ashfaq, Muhammad; Kiss, Zsofia; Khan, Azhar Abbas; Mansoor, Shahid; Falk, Bryce W

    2013-01-01

    The citrus mealybug, Planococcus citri, is an important plant pest with a very broad plant host range. P. citri is a phloem feeder and loss of plant vigor and stunting are characteristic symptoms induced on a range of host plants, but P. citri also reduces fruit quality and causes fruit drop leading to significant yield reductions. Better strategies for managing this pest are greatly needed. RNA interference (RNAi) is an emerging tool for functional genomics studies and is being investigated as a practical tool for highly targeted insect control. Here we investigated whether RNAi effects can be induced in P. citri and whether candidate mRNAs could be identified as possible targets for RNAi-based P. citri control. RNAi effects were induced in P. citri, as demonstrated by specific target reductions of P. citri actin, chitin synthase 1 and V-ATPase mRNAs after injection of the corresponding specific double-stranded RNA inducers. We also used recombinant Tobacco mosaic virus (TMV) to express these RNAi effectors in Nicotiana benthamiana plants. We found that P. citri showed lower fecundity and pronounced death of crawlers after feeding on recombinant TMV-infected plants. Taken together, our data show that actin, chitin synthase 1 and V-ATPase mRNAs are potential targets for RNAi against P. citri, and that recombinant TMV is an effective tool for evaluating candidate RNAi effectors in plants.

  15. [Analysis on medication rules of state medical master Yan Zhenghua from prescriptions with citri reticulatae pericarpium based on data mining].

    Science.gov (United States)

    Wu, Jia-Rui; Guo, Wei-Xian; Zhang, Bing; Zhang, Xiao-Meng; Yang, Bing; Sheng, Xiao-Guang

    2014-02-01

    The prescriptions containing pericarpium citri reticulatae that built by Professor. Yan were collected to build a database based on traditional Chinese medicine (TCM) inheritance assist system. After analyzed by data mining, such as apriori algorithm, the frequency of single medicine, the frequency of drug combination, the association rules between drugs and core drug combinations can be get from the database. Through the analysis of 1 027 prescriptions with pericarpium citri reticulatae, these prescriptions were commonly used to treat stomach aches, cough and other syndromes. The most frequency drug combinations were "Citri Reticulatae Pericarpium-Poria", "Paeoniae Radix Rubra-Citri Reticulatae Pericarpium" and so on. The drug association rules that the confidence was 1 were "Glycyrrhizae Radix ex Rhizoma --> Citri Reticulatae Pericarpium", "Paeoniae Alba Radix-Cyperi Rhizoma --> Citri Reticulatae Pericarpium", "Poria --> Citri Reticulatae Pericarpium", and so on. The drugs in the prescriptions containing pericarpium citri reticulatae that built by Professor Yan mostly had the effects of regulating the flow of Qi and invigorate blood circulation, which reflected the clearly thought when making prescriptions.

  16. Use of Electrical Penetration Graph Technology to Examine Transmission of 'Candidatus Liberibacter solanacearum' to Potato by Three Haplotypes of Potato Psyllid (Bactericera cockerelli; Hemiptera: Triozidae.

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    Tariq Mustafa

    Full Text Available The potato psyllid, Bactericera cockerelli (Šulc (Hemiptera: Triozidae, is a vector of the phloem-limited bacterium 'Candidatus Liberibacter solanacearum' (Lso, the putative causal agent of zebra chip disease of potato. Little is known about how potato psyllid transmits Lso to potato. We used electrical penetration graph (EPG technology to compare stylet probing behaviors and efficiency of Lso transmission of three haplotypes of potato psyllid (Central, Western, Northwestern. All haplotypes exhibited the full suite of stylet behaviors identified in previous studies with this psyllid, including intercellular penetration and secretion of the stylet pathway, xylem ingestion, and phloem activities, the latter comprising salivation and ingestion. The three haplotypes exhibited similar frequency and duration of probing behaviors, with the exception of salivation into phloem, which was of higher duration by psyllids of the Western haplotype. We manipulated how long psyllids were allowed access to potato ("inoculation access period", or IAP to examine the relationship between phloem activities and Lso transmission. Between 25 and 30% of psyllids reached and salivated into phloem at an IAP of 1 hr, increasing to almost 80% of psyllids as IAP was increased to 24 h. Probability of Lso-transmission was lower across all IAP levels than probability of phloem salivation, indicating that a percentage of infected psyllids which salivated into the phloem failed to transmit Lso. Logistic regression showed that probability of transmission increased as a function of time spent salivating into the phloem; transmission occurred as quickly as 5 min following onset of salivation. A small percentage of infected psyllids showed extremely long salivation events but nonetheless failed to transmit Lso, for unknown reasons. Information from these studies increases our understanding of Lso transmission by potato psyllid, and demonstrates the value of EPG technology in

  17. A Virulence-Reducing Mutation in the Postharvest Citrus Pathogen Alternaria citri.

    Science.gov (United States)

    Katoh, H; Isshiki, A; Masunaka, A; Yamamoto, H; Akimitsu, K

    2006-09-01

    ABSTRACT Alternaria citri causes Alternaria black rot, a postharvest fruit disease, on a broad range of citrus cultivars. We previously described that an endopolygalacturonase minus mutant of A. citri caused significantly less black rot in citrus fruit. To search for other essential factors causing symptoms in addition to endopolygalacturonase, a random mutation analysis of pathogenicity was performed using restriction enzyme-mediated integration. Three isolates among 1,694 transformants of A. citri had a loss in pathogenicity in a citrus peel assay, and one of these three mutants was a histidine auxotroph. Gene AcIGPD that encodes imidazole glycerol phosphate dehydratase, the sixth enzyme in the histidine biosynthetic pathway, was cloned, and the mutant containing the disrupted target gene, AcIGPD, caused less black rot.

  18. Use of Electrical Penetration Graph Technology to Examine Transmission of ‘Candidatus Liberibacter solanacearum’ to Potato by Three Haplotypes of Potato Psyllid (Bactericera cockerelli; Hemiptera: Triozidae)

    Science.gov (United States)

    The potato psyllid, Bactericera cockerelli (Šulc) (Hemiptera: Triozidae), is a vector of the phloem-limited bacterium ‘Candidatus Liberibacter solanacearum’ (Lso), the putative causal agent of zebra chip disease of potato. Little is known about how potato psyllid transmits Lso to potato. We used ele...

  19. Molecular detection of aster yellows phytoplasma and 'Candidatus Liberibacter solanacearum' in carrots affected by the psyllid Trioza apicalis (Hemiptera: triozidae) in Finland

    Science.gov (United States)

    Carrot psyllid (Trioza apicalis Förster) causes considerable damage to carrot (Daucus carota L.) in many parts of Europe. It was recently established that the new bacterium “Candidatus Liberibacter solanacearum” is associated with carrot psyllid and plants affected by this insect pest. No other path...

  20. Maintenance of primary cell cultures of immunocytes from Cacopsylla sp. psyllids: a new in vitrio tool for the study of pest insects

    Science.gov (United States)

    Psyllid species are major vectors of plant pathogens, such as phytoplasmas and Liberibacter bacteria, which threaten economic stability of fruit tee crops and vegetable production worldwide. Primary cell cultures of immunocytes have been developed from the three psyllid species, Cacopsylla melanone...

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

    Science.gov (United States)

    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...

  2. Geïntegreerde bestrijding van citruswolluis Planococcus citri in roos

    NARCIS (Netherlands)

    Pijnakker, J.; Leman, A.; Staaij, van der M.

    2011-01-01

    Referaat Sinds begin 2000 is citruswolluis, Planococcus citri (Risso), een van de belangrijkste plagen geworden in de rozenteelt onder glas. Telers kiezen voor een meer geïntegreerde aanpak om andere plagen onder controle te houden en stoppen met het reguliere gebruik van breedwerkende middelen. Hie

  3. 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 doorgaan

  4. Reduced Susceptibility to Xanthomonas citri in Transgenic Citrus Expressing the FLS2 Receptor From Nicotiana benthamiana.

    Science.gov (United States)

    Hao, Guixia; Pitino, Marco; Duan, Yongping; Stover, Ed

    2016-02-01

    Overexpression of plant pattern-recognition receptors by genetic engineering provides a novel approach to enhance plant immunity and broad-spectrum disease resistance. Citrus canker disease associated with Xanthomonas citri is one of the most important diseases damaging citrus production worldwide. In this study, we cloned the FLS2 gene from Nicotiana benthamiana cDNA and inserted it into the binary vector pBinPlus/ARS to transform Hamlin sweet orange and Carrizo citrange. Transgene presence was confirmed by polymerase chain reaction (PCR) and gene expression of NbFLS2 was compared by reverse transcription quantitative PCR. Reactive oxygen species (ROS) production in response to flg22Xcc was detected in transgenic Hamlin but not in nontransformed controls. Low or no ROS production was detected from nontransformed Hamlin seedlings challenged with flg22Xcc. Transgenic plants highly expressing NbFLS2 were selected and were evaluated for resistance to canker incited by X. citri 3213. Our results showed that the integration and expression of the NbFLS2 gene in citrus can increase canker resistance and defense-associated gene expression when challenged with X. citri. These results suggest that canker-susceptible Citrus genotypes lack strong basal defense induced by X. citri flagellin and the resistance of these genotypes can be enhanced by transgenic expression of the flagellin receptor from a resistant species.

  5. Antifungal Effect of the Aqueous and Ethanolic Avicennia marina Extracts on Alternaria citri and Penicillium digitatum

    Directory of Open Access Journals (Sweden)

    Behrooz Alizadeh Behbahani

    2016-02-01

    Full Text Available Background Plants have a great potential for producing new drugs of great benefit to mankind. There are many approaches to the search for new biologically active principles in higher plants. Objectives The aim of this study was to determine antifungal effect of the aqueous and ethanolic extracts of Avicennia marina on Penicillium digitatum and Alternaria citri in vitro. Materials and Methods In this experimental study, antimicrobial effects of extract were determined by screening antimicrobial activity and disk agar diffusion test in 20, 40, 60 and 80 mg/mL concentration of the extract against P. digitatum and A. citri. Minimum inhibitory concentration (MIC and minimum fungicidal concentration (MFC were determined by incorporating various concentrations of extracts in Sabouraud dextrose agar (SDA in tubes against P. digitatum and A. citri. The data were analyzed using one way analysis of variance (ANOVA using SPSS-17. Results The results indicate that extracts of A. marina mostly had been effective on P. digitatum and have the least impact on A. citri. The results show that the ethanol extract of mangrove leaf with antimicrobial activity method in 2 mg/mL, were inhibited the growth of P. digitatum and A. citri. In disk agar diffusion test the mentioned extract were shown inhibition effect on pathogen molds growth (P < 0.05. Conclusions Ethanolic extract compared to the aqueous extract was more effective and has a greater inhibition effect. The results of this study showed that the extract of A. marina has antifungal properties that justify their traditional use as medicinal plants.

  6. Expressão gênica diferencial de laranja Pêra Rio (Citrus sinensis (L.) Osbeck) e Lima Ácida 'Galego' (Citrus aurantifolia Swingle) em resposta à infecção por Xanthomonas citri subsp. citri

    OpenAIRE

    Cavallini, Juliana da Silva [UNESP

    2013-01-01

    The citrus agribusiness is very important to the Brazilian economy, but the increase of diseases in the last decade has caused great economic losses to the sector. The citrus canker, caused by the bacterium Xanthomonas citri subsp. citri (Xac), is a serious disease that attacks all citrus species economically important worldwide and there is not an effective method for its control. In this study, RNASeq was used to analyze the transcriptional profiles of two contrasting citrus genotypes regar...

  7. Enhanced Acquisition Rates of 'Candidatus Liberibacter asiaticus' by the Asian Citrus Psyllid (Hemiptera: Liviidae) in the Presence of Vegetative Flush Growth in Citrus.

    Science.gov (United States)

    Sétamou, Mamoudou; Alabi, Olufemi J; Kunta, Madhurababu; Jifon, John L; da Graça, John V

    2016-10-01

    The Asian citrus psyllid preferentially feeds and exclusively reproduces on young, newly emerged flush shoots of citrus. Asian citrus psyllid nymphs feed and complete their life stages on these flush shoots. Recent studies conducted under greenhouse conditions have shown that the transmission rates of 'Candidatus Liberibacter asiaticus' (CLas), the putative causal agent of huanglongbing disease of citrus, are enhanced when flush shoots are present. However, it is unclear if CLas acquisition by migrant adult Asian citrus psyllids is similarly enhanced. To address this knowledge gap, cohorts of Asian citrus psyllid adults were allowed 1-wk acquisition access period (AAP) on flushing and nonflushing shoots of qPCR-tested symptomatic (CLas+) and asymptomatic (CLas-) 10-yr-old sweet orange trees under field conditions. After the AAP, they were tested for CLas by qPCR. Progeny Asian citrus psyllid adults that emerged 4 wk post-AAP were similarly retrieved and tested. Eighty percent of flushing and 30% of nonflushing CLas+ trees produced infective Asian citrus psyllid adults, indicating that flush shoots have greater potential to be inoculum sources for CLas acquisition. Concomitantly, 21.1% and 6.0% infective adults were retrieved, respectively, from flushing and nonflushing CLas+ trees, indicating that Asian citrus psyllid adults acquire CLas more efficiently from flush shoots relative to mature shoots. In addition, 12.1% of infective Asian citrus psyllid adult progeny were obtained from 70% of flushing CLas+ trees. Significantly lower mean Ct values were also obtained from infective adults retrieved from flushing relative to nonflushing trees. The results underscore the role of flush shoots in CLas acquisition and the need to protect citrus trees from Asian citrus psyllid infestations during flush cycles.

  8. Zebra chip disease incidence on potato is influenced by timing of potato psyllid infestation,but not by the host plants on which they were reared

    Institute of Scientific and Technical Information of China (English)

    Feng Gao; John Jifon; Xiangbing Yang; Tong-Xian Liu

    2009-01-01

    The Zebra chip (ZC) syndrome is an emerging disease of potato and a major threat to the potato industry.The potato psyllid,Bactericerca cockerelli (Sulc) is believed to be a vector of the ZC pathogen,which is now thought to be Candidatus Liberibacter,a bacterium.To further understand the relationship between potato psyllid infestation and ZC disease expression,healthy potato plants at different growth stages (4,6 and 10 weeks after germination) were exposed separately to potato psyllids that were separately reared on four solanaceons hosts plants (potato,tomato,eggplant or bell pepper) for more than 1 year.ZC symptoms,leaf rates and total nonstructural carbohydrate accumulation in leaves and tubers of healthy and psyllid-infested plants were monitored and recorded.Typical ZC symptoms were observed in leaves and tubers of all plants exposed to potato psyllids regardless of the host plant on which they were reared.This was also accompanied by significant reductions in net photosynthetic rate.Caged potato plants without exposure to potato psyllids (uninfested controls) did not show any ZC symptom in both foliage and in harvested tubers.Foliage damage and ZC expression were most severe in the potato plants that were exposed to potato psyllids 4 weeks after germination compared to plants infested at later growth stages.Tubers from potato psyllid-infested plants had significantly higher levels of reducing sugars (glucose) and lower levels of starch than those in healthy plants,indicating that potato psyllid infestation interfered with carbohydrate metabolism in either leaves or tubers,resulting in ZC expression.

  9. Green Fluorescent Detection of Fungal Colonization and Endopolygalacturonase Gene Expression in the Interaction of Alternaria citri with Citrus.

    Science.gov (United States)

    Isshiki, A; Ohtani, K; Kyo, M; Yamamoto, H; Akimitsu, K

    2003-07-01

    ABSTRACT Alternaria citri, a postharvest pathogen, produces endopolygalacturonase (endoPG) and causes black rot on citrus fruit. We previously described that an endoPG-disrupted mutant of Alternaria citri was significantly reduced in its ability to macerate plant tissue and cause black rot symptoms on citrus. In order to investigate colonization of citrus fruit tissues by Alternaria citri, pTEFEGFP carrying a green fluorescent protein (GFP) gene was introduced into wild-type Alternaria citri and its endoPG-disrupted mutant (M60). Green fluorescence was observed in spores, germ tubes, appressoria, and infection hyphae of transformants G1 (derived from wild type) and GM4 (derived from M60). Hyphae of G1 but not GM4 vertically penetrated the peel, but the hyphae of both G1 and GM4 spread equally in the juice sac area of citrus fruit. Green fluorescence of Alternaria citri transformant EPG7 carrying a GFP gene under control of the endoPG gene promoter of Alternaria citri was induced by pectin in the peel during the infection stage, but repressed completely in the juice sac area, likely by carbon catabolite repression by sugars in the juice.

  10. Survey of 'Candidatus Liberibacter solanacearum' in carrot crops affected by the psyllid Trioza apicalis (Hemiptera: Triozidae) in Norway

    Science.gov (United States)

    The carrot psyllid Trioza apicalis Förster (Hemiptera: Triozidae) is a serious insect pest of carrot (Daucus carota L.) in northern Europe, where it can cause up to 100% crop loss. Although it was long believed that T. apicalis causes damage to carrot by injection of toxins into the plant, it was re...

  11. Volatile profiles of young leaves of Rutaceae spp. varying in susceptibility to the Asian citrus psyllid,(Hemiptera: Psyllidae)

    Science.gov (United States)

    Plant volatiles were identified from six species in the family Rutaceae. These species had varying degrees of susceptibility to the Asian citrus psyllid as determined by direct counts of life stages. Using a push system involving charcoal-filtered humidified air, volatiles were adsorbed on SuperQ pa...

  12. New corological and biological data of the Red Gum Lerp Psyllid, Glycaspis brimblecombei Moore, 1964 in Italy (Hemiptera, Psyllidae

    Directory of Open Access Journals (Sweden)

    Ricardo Jiménez-Peydró

    2011-03-01

    Full Text Available Glycaspis brimblecombei Moore, 1964 is a psyllid (Hemiptera: Psyllidae pest of Eucalyptus, native toAustralia and first recorded in Europe: Spain in 2008 and more recently (2010 in Italy. The present paper dealswith recent research, carried out in central Italy, with new data on the distribution and biology of this species.

  13. Repellency of a kaolin particle film to potato psyllid, Bactericera cockerelli (Hemiptera: Psyllidae), on tomato under laboratory and field conditions.

    Science.gov (United States)

    Peng, Linian; Trumble, John T; Munyaneza, Joseph E; Liu, Tong-Xian

    2011-07-01

    The potato psyllid, Bactericera cockerelli, is a vector of Candidatus Liberibacter solanacearum, causing several diseases in solanaceous crops. Laboratory and field no-choice and choice experiments were conducted to evaluate the repellency of kaolin particle film on adults of B. cockerelli on tomato plants that had been sprayed with kaolin particle film on the upper surface only, on the lower surface only and on both leaf surfaces. In no-choice tests in the laboratory, the numbers of adults on leaves were not different between the kaolin particle film and the water control, regardless of which leaf surface(s) were treated, but numbers of eggs were lower on the leaves treated with kaolin particle film than on those treated with water. In choice tests on plants treated with water/plants treated with kaolin particle film at ratios of 1:1, 6:3 or 8:1, fewer adults and eggs were found on the leaves treated with kaolin particle film than on leaves treated with water. Under field conditions, in caged no-choice or choice tests, fewer adults, eggs and nymphs were found on plants treated with kaolin particle film than on plants treated with water. In an uncaged test under field conditions, plants sprayed with kaolin particle film had fewer psyllids than those sprayed with water. Even though potato psyllid adults could land on plants treated with kaolin particle film when no choice was given, fewer eggs were laid. When given a choice, the psyllids avoided plants treated with kaolin particle film under laboratory and field conditions. Kaolin particle film treatment may be a useful alternative for management of potato psyllids under field conditions. Copyright © 2011 Society of Chemical Industry.

  14. Differential responses to feeding by the tomato/potato psyllid between two tomato cultivars and their implications in establishment of injury levels and potential of damaged plant recovery

    Institute of Scientific and Technical Information of China (English)

    DEGUANG LIU; LINDSEY JOHNSON; JOHN T. TRUMBLE

    2006-01-01

    An invasive new biotype of the tomato/potato psyllid (Bactericera [Paratrioza] cockerelli [Sulc.]) (Homoptera: Psyllidae) recently has caused losses exceeding 50% on fresh market tomatoes in western North America. Despite these extensive losses, little is known regarding the threshold levels at which populations must be suppressed in order to prevent economic losses. A series of experiments were therefore designed using combinations of two common tomato cultivars (QualiT 21 and Yellow Pear), five pest-densities (0, 20, 30, 40 and 50 nymphs/plant), and three feeding-duration (5 days, 10 days, and lifetime) treatments to test the relative importance of pest density, feeding period, and cumulative psyllid-days to establish economic threshold levels for psyllids. The cultivars differed considerably in their response to the toxin injected by the psyllid nymphs. 'Yellow Pear' plants could recover from feeding by up to 40 nymphs for as long as 10 d, whereas 'QualiT 21' plants were irreparably damaged by densities of 20 nymphs feeding for only 5 days. On 'Yellow Pear', all plant measurements such as the number of yellow leaves and plant height were significantly better correlated with cumulative psyllid-days than with either pest density or feeding duration. On 'QualiT 21', all plant measurements other than the number of yellow leaflets and leaves were significantly better correlated with pest density than with feeding duration or cumulative psyllid-days, and pest density was a better predictor of psyllid damage. Potential reasons for the variable responses between cultivars and the implications for psyllid sampling and integrated pest management are discussed.

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

    Energy Technology Data Exchange (ETDEWEB)

    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)

  16. Novel diagnosis for citrus stubborn disease by detection of a spiroplasma citri-secreted protein.

    Science.gov (United States)

    Shi, Jinxia; Pagliaccia, Deborah; Morgan, Robyn; Qiao, Yongli; Pan, Songqin; Vidalakis, Georgios; Ma, Wenbo

    2014-02-01

    Citrus stubborn disease (CSD), first identified in California, is a widespread bacterial disease found in most arid citrus-producing regions in the United States and the Mediterranean Region. The disease is caused by Spiroplasma citri, an insect-transmitted and phloem-colonizing bacterium. CSD causes significant tree damage resulting in loss of fruit production and quality. Detection of CSD is challenging due to low and fluctuating titer and sporadic distribution of the pathogen in infected trees. In this study, we report the development of a novel diagnostic method for CSD using an S. citri-secreted protein as the detection marker. Microbial pathogens secrete a variety of proteins during infection that can potentially disperse systemically in infected plants with the vascular flow. Therefore, their distribution may not be restricted to the pathogen infection sites and could be used as a biological marker for infection. Using mass spectrometry analysis, we identified a unique secreted protein from S. citri that is highly expressed in the presence of citrus phloem extract. ScCCPP1, an antibody generated against this protein, was able to distinguish S. citri-infected citrus and periwinkle from healthy plants. In addition, the antiserum could be used to detect CSD using a simple direct tissue print assay without the need for sample processing or specialized lab equipment and may be suitable for field surveys. This study provides proof of a novel concept of using pathogen-secreted protein as a marker for diagnosis of a citrus bacterial disease and can probably be applied to other plant diseases.

  17. Adjuvants to improve aerial control of the citrus mealybug Planococcus citri (Hemiptera: Pseudococcidae) using entomopathogenic nematodes.

    Science.gov (United States)

    van Niekerk, S; Malan, A P

    2015-03-01

    The citrus mealybug, Planococcus citri, is a highly destructive pest of citrus, occurring only in the aerial parts of plants. Humidity will be one of the key factors to consider when using entomopathogenic nematodes (EPN) as biological control agents. Different adjuvants can be added to suspensions of EPNs, to improve control as a foliar application. An aqueous suspension containing Heterorhabditis zealandica and 0.3% Zeba® significantly increased P. citri mortality by 22% at 80% relative humidity (RH) with a temperature cycle starting at 22°C for 14 h and 11°C for 11 h. The same polymer formulation was tested for Steinernema yirgalemense and mortality of P. citri increased by 21% at 60% RH and by 27% at 80% RH. The addition of Nu-Film-P® and Zeba® to H. zealandica suspensions did not significantly retard application run-off on citrus leaves. The combination of Nu-Film-P® and Zeba®, however, was able to significantly retard sedimentation, increasing the average number of nematodes deposited on 2-cm2 leaf discs by 10 nematodes. In an aqueous suspension, nematodes settle rapidly to the bottom, resulting in an uneven distribution of nematodes. Xanthan gum, at a concentration of 0.2%, was highly effective at retarding sedimentation, with 72% of the initial nematode number still in suspension after 1 h. Zeba®, at a concentration of 0.3%, despite not being as effective as Xanthan gum, nevertheless still retarded sedimentation significantly. This is the first report of the potential of Nu-Film-P® and Zeba® to improve EPN performance against P. citri when used above ground in citrus orchards.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    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.

  1. Evolutionary conservation of candidate osmoregulation genes in plant phloem sap-feeding insects.

    Science.gov (United States)

    Jing, X; White, T A; Luan, J; Jiao, C; Fei, Z; Douglas, A E

    2016-06-01

    The high osmotic pressure generated by sugars in plant phloem sap is reduced in phloem-feeding aphids by sugar transformations and facilitated water flux in the gut. The genes mediating these osmoregulatory functions have been identified and validated empirically in the pea aphid Acyrthosiphon pisum: sucrase 1 (SUC1), a sucrase in glycoside hydrolase family 13 (GH13), and aquaporin 1 (AQP1), a member of the Drosophila integral protein (DRIP) family of aquaporins. Here, we describe molecular analysis of GH13 and AQP genes in phloem-feeding representatives of the four phloem-feeding groups: aphids (Myzus persicae), coccids (Planococcus citri), psyllids (Diaphorina citri, Bactericera cockerelli) and whiteflies (Bemisia tabaci MEAM1 and MED). A single candidate GH13-SUC gene and DRIP-AQP gene were identified in the genome/transcriptome of most insects tested by the criteria of sequence motif and gene expression in the gut. Exceptionally, the psyllid Ba. cockerelli transcriptome included a gut-expressed Pyrocoelia rufa integral protein (PRIP)-AQP, but has no DRIP-AQP transcripts, suggesting that PRIP-AQP is recruited for osmoregulatory function in this insect. This study indicates that phylogenetically related SUC and AQP genes may generally mediate osmoregulatory functions in these diverse phloem-feeding insects, and provides candidate genes for empirical validation and development as targets for osmotic disruption of pest species. © 2016 The Authors. Insect Molecular Biology published by John Wiley & Sons Ltd on behalf of The Royal Entomological Society.

  2. Organization of the mitochondrial genomes of whiteflies, aphids, and psyllids (Hemiptera, Sternorrhyncha)

    Science.gov (United States)

    Thao, MyLo L; Baumann, Linda; Baumann, Paul

    2004-01-01

    Background With some exceptions, mitochondria within the class Insecta have the same gene content, and generally, a similar gene order allowing the proposal of an ancestral gene order. The principal exceptions are several orders within the Hemipteroid assemblage including the order Thysanoptera, a sister group of the order Hemiptera. Within the Hemiptera, there are available a number of completely sequenced mitochondrial genomes that have a gene order similar to that of the proposed ancestor. None, however, are available from the suborder Sternorryncha that includes whiteflies, psyllids and aphids. Results We have determined the complete nucleotide sequence of the mitochondrial genomes of six species of whiteflies, one psyllid and one aphid. Two species of whiteflies, one psyllid and one aphid have mitochondrial genomes with a gene order very similar to that of the proposed insect ancestor. The remaining four species of whiteflies had variations in the gene order. In all cases, there was the excision of a DNA fragment encoding for cytochrome oxidase subunit III(COIII)-tRNAgly-NADH dehydrogenase subunit 3(ND3)-tRNAala-tRNAarg-tRNAasn from the ancestral position between genes for ATP synthase subunit 6 and NADH dehydrogenase subunit 5. Based on the position in which all or part of this fragment was inserted, the mitochondria could be subdivided into four different gene arrangement types. PCR amplification spanning from COIII to genes outside the inserted region and sequence determination of the resulting fragments, indicated that different whitefly species could be placed into one of these arrangement types. A phylogenetic analysis of 19 whitefly species based on genes for mitochondrial cytochrome b, NADH dehydrogenase subunit 1, and 16S ribosomal DNA as well as cospeciating endosymbiont 16S and 23S ribosomal DNA indicated a clustering of species that corresponded to the gene arrangement types. Conclusions In whiteflies, the region of the mitochondrial genome

  3. Organization of the mitochondrial genomes of whiteflies, aphids, and psyllids (Hemiptera, Sternorrhyncha

    Directory of Open Access Journals (Sweden)

    Baumann Paul

    2004-08-01

    Full Text Available Abstract Background With some exceptions, mitochondria within the class Insecta have the same gene content, and generally, a similar gene order allowing the proposal of an ancestral gene order. The principal exceptions are several orders within the Hemipteroid assemblage including the order Thysanoptera, a sister group of the order Hemiptera. Within the Hemiptera, there are available a number of completely sequenced mitochondrial genomes that have a gene order similar to that of the proposed ancestor. None, however, are available from the suborder Sternorryncha that includes whiteflies, psyllids and aphids. Results We have determined the complete nucleotide sequence of the mitochondrial genomes of six species of whiteflies, one psyllid and one aphid. Two species of whiteflies, one psyllid and one aphid have mitochondrial genomes with a gene order very similar to that of the proposed insect ancestor. The remaining four species of whiteflies had variations in the gene order. In all cases, there was the excision of a DNA fragment encoding for cytochrome oxidase subunit III(COIII-tRNAgly-NADH dehydrogenase subunit 3(ND3-tRNAala-tRNAarg-tRNAasn from the ancestral position between genes for ATP synthase subunit 6 and NADH dehydrogenase subunit 5. Based on the position in which all or part of this fragment was inserted, the mitochondria could be subdivided into four different gene arrangement types. PCR amplification spanning from COIII to genes outside the inserted region and sequence determination of the resulting fragments, indicated that different whitefly species could be placed into one of these arrangement types. A phylogenetic analysis of 19 whitefly species based on genes for mitochondrial cytochrome b, NADH dehydrogenase subunit 1, and 16S ribosomal DNA as well as cospeciating endosymbiont 16S and 23S ribosomal DNA indicated a clustering of species that corresponded to the gene arrangement types. Conclusions In whiteflies, the region of the

  4. Xanthomonas citri subsp. citri surface proteome by 2D-DIGE: Ferric enterobactin receptor and other outer membrane proteins potentially involved in citric host interaction.

    Science.gov (United States)

    Carnielli, Carolina Moretto; Artier, Juliana; de Oliveira, Julio Cezar Franco; Novo-Mansur, Maria Teresa Marques

    2017-01-16

    Xanthomonas citri subsp. citri (XAC) is the causative agent of citrus canker, a disease of great economic impact around the world. Understanding the role of proteins on XAC cellular surface can provide new insights on pathogen-plant interaction. Surface proteome was performed in XAC grown in vivo (infectious) and in vitro (non-infectious) conditions, by labeling intact cells followed by cellular lysis and direct 2D-DIGE analysis. Seventy-nine differential spots were analyzed by mass spectrometry. Highest relative abundance for in vivo condition was observed for spots containing DnaK protein, 60kDa chaperonin, conserved hypothetical proteins, malate dehydrogenase, phosphomannose isomerase, and ferric enterobactin receptors. Elongation factor Tu, OmpA-related proteins, Oar proteins and some Ton-B dependent receptors were found in spots decreased in vivo. Some proteins identified on XAC's surface in infectious condition and predicted to be cytoplasmic, such as DnaK and 60KDa chaperonin, have also been previously found at cellular surface in other microorganisms. This is the first study on XAC surface proteome and results point to mediation of molecular chaperones in XAC-citrus interaction. The approach utilized here can be applied to other pathogen-host interaction systems and help to achieve new insights in bacterial pathogenicity toward promising targets of biotechnological interest.

  5. Potential of South African entomopathogenic nematodes (Heterorhabditidae and Steinernematidae) for control of the citrus mealybug, Planococcus citri (Pseudococcidae).

    Science.gov (United States)

    van Niekerk, Sonnica; Malan, Antoinette P

    2012-10-01

    Planococcus citri, the citrus mealybug, is the most important species of mealybug known to infest citrus in South Africa. Various laboratory bioassays were conducted to determine the potential of entomopathogenic nematodes to control P. citri. Adult female P. citri were screened for susceptibility to six indigenous nematode species. P. citri was found to be most susceptible to Steinernema yirgalemense and Heterorhabditis zealandica, causing 97% and 91% mortality, respectively. The development of nematodes after infecting adult female P. citri showed both H. zealandica and S. yirgalemense were able to complete their life cycles inside the host. Further bioassays illustrated a linear relationship between mealybug mortality and the concentration of nematodes applied, with the highest level of control using a concentration of 80 infective juveniles (IJs)/insect. As nematodes would be used as an above-ground application to control P. citri in citrus orchards, available water is a major limiting factor. Insecticidal activity proved to be dependent on the available surface moisture after nematode application. The water activity (a(w)) bioassay indicated that S. yirgalemense to be two times more tolerant to lower levels of free water, with a(w50)=0.96 and a(w90)=0.99, compared to H. zealandica with a(w50)=0.98 and a(w)90=1.0. After application, nematodes have a limited time frame in which to locate and infect hosts, as the level of available free water gradually decreases, as trees dry out. S. yirgalemense proved able to locate and infect P. citri quicker than H. zealandica. Nematode activity was not significantly affected when exposed to 15°C, 20°C and 25°C. IJs were able to infect P. citri at an exposure time as short as half an hour. Results also showed that the first 2-4h post application is the most decisive time for establishing successful infection of mealybugs. This is the first report on the potential use of nematodes for the control of P. citri.

  6. Analyses of Numerical Responses and Main Life Parameters for Determining the Suppression of Amblyseius cucumeris on Panonychus citri

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yan-xuan; LIN Jian-zhen; JI Jie; CHEN Xie; KANG Yu-mei

    2005-01-01

    In this paper, the numeral response and main parameters of experimental population life table were analyzed for determiningthe suppressing ability of Amblyseius cucumeris on Panonychus citri. The result showed that: (1) Under 21-31 ℃ and 1-9 prey densities/leaf fragment condition, the prey consumptions ofA. cucumeris increased with the temperature or prey density; (2) In 1:3 predator-prey ratio treatment, the suppression ofP. citri (Ro=34.0053; T=19.4369; t=3.8204) was rather weak, it was enhanced as the ratio over 5:30, and the populations ofP. citri in these treatments can be fully controlled within 4-5 days; (3) Under 25±1℃, 80-85% RH and 15L: 9D illumination conditions, the net reproductive rate, mean generation duration and the time for population double increase ofP. citri (Ro=34.0053; T=19.4369; t=3.8204) were higher than those ofA. cucumeris (Ro=21.8750; T=16.8943; t=3.7954). While the intrinsic rate of increase and finite rate of increase of the former (rm=0.1814; λ= 1.1989) were lower than those of the latter (rm=0.1826; λ= 1.2004). These results indicated that A. cucumeris is a desirable bio-control agent to suppress P. citri at lower population stage in citrus orchard.

  7. Expression patterns of genes involved in the defense and stress response of Spiroplasma citri infected Madagascar Periwinkle Catharanthus roseus.

    Science.gov (United States)

    Nejat, Naghmeh; Vadamalai, Ganesan; Dickinson, Matthew

    2012-01-01

    Madagascar periwinkle is an ornamental and a medicinal plant, and is also an indicator plant that is highly susceptible to phytoplasma and spiroplasma infections from different crops. Periwinkle lethal yellows, caused by Spiroplasma citri, is one of the most devastating diseases of periwinkle. The response of plants to S. citri infection is very little known at the transcriptome level. In this study, quantitative real-time PCR (RT-qPCR) was used to investigate the expression levels of four selected genes involved in defense and stress responses in naturally and experimentally Spiroplasma citri infected periwinkles. Strictosidine β-glucosidase involved in terpenoid indole alkaloids (TIAs) biosynthesis pathway showed significant upregulation in experimentally and naturally infected periwinkles. The transcript level of extensin increased in leaves of periwinkles experimentally infected by S. citri in comparison to healthy ones. A similar level of heat shock protein 90 and metallothionein expression was observed in healthy, naturally and experimentally spiroplasma-diseased periwinkles. Overexpression of Strictosidine β-glucosidase demonstrates the potential utility of this gene as a host biomarker to increase the fidelity of S. citri detection and can also be used in breeding programs to develop stable disease-resistance varieties.

  8. Expression Patterns of Genes Involved in the Defense and Stress Response of Spiroplasma citri Infected Madagascar Periwinkle Catharanthus roseus

    Directory of Open Access Journals (Sweden)

    Naghmeh Nejat

    2012-02-01

    Full Text Available Madagascar periwinkle is an ornamental and a medicinal plant, and is also an indicator plant that is highly susceptible to phytoplasma and spiroplasma infections from different crops. Periwinkle lethal yellows, caused by Spiroplasma citri, is one of the most devastating diseases of periwinkle. The response of plants to S. citri infection is very little known at the transcriptome level. In this study, quantitative real-time PCR (RT-qPCR was used to investigate the expression levels of four selected genes involved in defense and stress responses in naturally and experimentally Spiroplasma citri infected periwinkles. Strictosidine β-glucosidase involved in terpenoid indole alkaloids (TIAs biosynthesis pathway showed significant upregulation in experimentally and naturally infected periwinkles. The transcript level of extensin increased in leaves of periwinkles experimentally infected by S. citri in comparison to healthy ones. A similar level of heat shock protein 90 and metallothionein expression was observed in healthy, naturally and experimentally spiroplasma-diseased periwinkles. Overexpression of Strictosidine β-glucosidase demonstrates the potential utility of this gene as a host biomarker to increase the fidelity of S. citri detection and can also be used in breeding programs to develop stable disease-resistance varieties.

  9. Candidate gene markers for Candidatus Liberibacter asiaticus for detecting citrus greening disease.

    Science.gov (United States)

    Nageswara-Rao, Madhugiri; Irey, Mike; Garnsey, Stephen M; Gowda, Siddarame

    2013-06-01

    Citrus Huanglongbing (HLB) also known as citrus greening is one of the most devastating diseases of citrus worldwide. The disease is caused by Candidatus Liberibacter bacterium, vectored by the psyllid Diaphorina citri Kuwayama and Trioza erytreae Del Guercio. Citrus plants infected by the HLB bacterium may not show visible symptoms sometimes for years following infection. The aim of this study was to develop effective gene-specific primer pairs for polymerase chain reaction based method for quick screening of HLB disease. Thirty-two different gene-specific primer pairs, across the Ca. Liberibacter asiaticus genome, were successfully developed. The possibility of these primer pairs for cross-genome amplification across 'Ca. Liberibacter africanus' and 'Ca. Liberibacter americanus' were tested. The applicability of these primer pairs for detection and differentiation of Ca Liberibacter spp. is discussed.

  10. Candidate gene markers for Candidatus Liberibacter asiaticus for detecting citrus greening disease

    Indian Academy of Sciences (India)

    Madhugiri Nageswara-Rao; Mike Irey; Stephen M Garnsey; Siddarame Gowda

    2013-06-01

    Citrus Huanglongbing (HLB) also known as citrus greening is one of the most devastating diseases of citrus worldwide. The disease is caused by Candidatus Liberibacter bacterium, vectored by the psyllid Diaphorina citri Kuwayama and Trioza erytreae Del Guercio. Citrus plants infected by the HLB bacterium may not show visible symptoms sometimes for years following infection. The aim of this study was to develop effective gene-specific primer pairs for polymerase chain reaction based method for quick screening of HLB disease. Thirty-two different gene-specific primer pairs, across the Ca. Liberibacter asiaticus genome, were successfully developed. The possibility of these primer pairs for cross-genome amplification across `Ca. Liberibacter africanus’ and `Ca. Liberibacter americanus’ were tested. The applicability of these primer pairs for detection and differentiation of Ca Liberibacter spp. is discussed.

  11. Establishment of EMA-PCR detection method of viable cells of Xanthomonas citri subsp.citri%柑橘溃疡病菌EMA-PCR快速活体检测技术的建立

    Institute of Scientific and Technical Information of China (English)

    熊书; 王中康; 卢小林; 殷幼平

    2013-01-01

    传统PCR方法不能诊断柑橘溃疡病菌(Xanthomonas citri subsp.citri Gabriel)的死活状态,往往导致假阳性检测结果.本研究将特异性核酸染料叠氮溴化乙锭(ethidium monoazide bromide,EMA)与PCR技术结合,旨在建立柑橘溃疡病活菌的快速检测技术.根据柑橘溃疡病菌独有的保守蛋白基因设计特异性引物扩增出278 bp的靶带,PCR反应的检测下限为25个细胞/25 μL或2.75 pg/25 μL.EMA-PCR结果表明:当卤钨灯曝光时间1 min,EMA终浓度为1.0 mg/L时,能有效抑制1.0×108 cfu/mL死菌的扩增;当EMA的浓度小于30 mg/L时,EMA对上述相同浓度活菌靶基因的扩增没有明显的抑制.EMA-PCR对死活混合菌的扩增表明,活菌数在6.875×101~6.875×105 cfu/PCR范围时,荧光强度与混合体系中活菌的对数值有线性关系.基于以上建立的EMA-PCR活体检测技术,对疑似带病柑橘材料进行检测,结果发现能降低柑橘溃疡病菌检测过程中的假阳性,有望为柑橘溃疡病的检疫检验提供更科学的技术手段.%Conventional PCR method can not distinguish live cells from dead cells of Xanthononas citri subsp.citri Gabriel,and false positives results can easily be obtained in detection process.Ethidium monoazide bromide (EMA) can selectively inhibit PCR amplification of DNA from dead cells.Therefore,in this study,EMA was combined with PCR to detect viable cells of X.citri subsp.citri.Firstly,primers (Xcc R/Xcc F) specific to X.citri subsp.citri were designed to amplify a 278 bp fragment.The minimum limit of detection was 25 cells/25 μL PCR volume or 2.75 pg/25 μ L PCR volume.The results of EMA-PCR showed that the optimized light exposure time was at least 1 min,allowing crosslinking of DNA by the EMA in dead cells and photolysing the free EMA in solution.The minimum amount of EMA to completely inhibit the PCR amplification of DNA derived from heat-killed cells was 1.0 mg/L.EMA less than 30 mg/L did not inhibit the PCR amplification

  12. Field Experiments of 13 Pesticides for Control Asian Citrus Psylla(Diaphorina citri Kuwayama)%吡蚜酮等13种农药防治柑桔木虱田间药效试验

    Institute of Scientific and Technical Information of China (English)

    邓明学; 唐美欣; 谭有龙; 陈贵峰; 唐明丽

    2009-01-01

    柑桔黄龙病是世界性的毁灭性病害,柑桔木虱是其唯一自然传播媒介,控制柑桔木虱是柑桔黄龙病综合防治最重要的技术措施.柑桔木虱成虫的高效传病性、近距离扩散性和快速传病性,决定了用来防治柑桔木虱成虫的农药要求速效性好、防效高(药后24h的防效>95%).由于受到试验地虫源和试验条件的限制,柑桔木虱的有效防治药剂筛选严重滞后.2007-2008年做了13种农药的田间小区药效试验和3种农药的大面积应用试验,结果表明:增效联苯菊酯EC 25 mg(a.i)/L、雕杀EC 110 mg(a.i)/L、铁骑EC 83 mg(a.i)/L对柑桔木虱成虫药后24 h的防效达100%;喹硫磷EC 250 mg(a.i)/L、杀扑磷EC200 mg(a.i)/L、稻丰散EC 333 mg(a.i)/L对柑桔木虱成虫药后24 h的防效>95%;防效在94.9%~85%的有丁硫克百威EC 200 mg(a.i)/L、霸中霸EC 200 mg(a.i)/L;吡虫啉WP 100 mg(a.i)/L、阿维·丁硫EC 90 mg(a.i)/L、甲基嘧啶磷500mg(a.i)/L的防效分别为92.3%~74.4%、79.1%、73.1;防效低于40%的药剂有吡蚜酮WP 250mg(a.i)/L、印楝素EC 6 mg(a.i)/L.

  13. Field Experiments of 15 Pesticides for Control Asian Citrus Psylla (Diaphorina citri Kuwayama)%20%哒虱威乳油等15种农药防治柑桔木虱田间药效试验

    Institute of Scientific and Technical Information of China (English)

    邓明学; 石晓雨

    2007-01-01

    控制柑桔木虱是柑桔黄龙病综合防治最重要的技术措施.柑桔木虱成虫的高效传病性、近距离扩散性、终生带毒性和快速传病性,决定了用来防治柑桔木虱成虫的农药要求速效性好、防效高(药后24 h的防效>95%).由于受到试验地虫源和试验条件的限制,柑桔木虱的有效防治药剂筛选严重滞后.2006-2007年做了15种农药的田间药效试验,结果表明:哒虱威EC 200mg(a.i)/L、丁硫克百威EC400mg(a.i)/L对柑桔木虱成虫药后24h的防效>95%;防效在94.9%~90%的有异丙威200mg(a.i)/L、133mg(a.i)/L、丁硫克百威EC 250mg(a.i)/L;噻嗪酮250mg(a.i)/L的防效只有30.8%;防效低于30%的药剂有灭多威200mg(a.i)/L、鱼藤酮150mg(a.i)/L、机油乳剂950mg(a.i)/L、阿维菌素20mg(a.i)/L和石硫合剂1波美度.机油乳剂与哒虱威混用无增效作用.

  14. 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.

  15. Evaluation of efficacy of 18 strains of entomopathogenic nematodes (Rhabditida) against Planococcus citri (Risso, 1813) (Hemiptera: Pseudococcidae) under laboratory conditions.

    Science.gov (United States)

    Barbosa Negrisoli, Carla Ruth de Carvalho; Negrisoli Júnior, Aldomario Santo; Botton, Marcos; Garcia, Mauro Silveira; Bernardi, Daniel

    2013-07-01

    Planococcus citri (Risso, 1813) (Hemiptera: Pseudococcidae) is an important plant virus vector in grapevine crops in Brazil and other countries. The mealybug grows in roots and leaves of the grapes. Entomopathogenic nematodes (EPNs) are efficient control agents against insects associated to the soil and could be applied with the same equipment used for chemical insecticides. The aim of this study was to select effective EPNs for controlling P. citri females in laboratory conditions (25±1°C, UR 60±10%). We tested 17 native [Steinernema rarum (6 strains), Steinernema glaseri, Steinernema feltiae, Steinernema riobrave, Steinernema sp., Heterorhabditis bacteriophora (7 strains)] and only one exotic strain (Steinernema carpocapsae ALL). The bioassays were done on Petri dishes infested with females of P. citri, which were sprayed with EPNs juveniles. The strain with larger pathogenicity and virulence in laboratory was H. bacteriophora RS33 (from 69.0% to 92.2% of mortality), native of Rio Grande do Sul.

  16. Structural and Physiological Analyses of the Alkanesulphonate-Binding Protein (SsuA) of the Citrus Pathogen Xanthomonas citri

    Science.gov (United States)

    Tófoli de Araújo, Fabiano; Bolanos-Garcia, Victor M.; Pereira, Cristiane T.; Sanches, Mario; Oshiro, Elisa E.; Ferreira, Rita C. C.; Chigardze, Dimitri Y.; Barbosa, João Alexandre Gonçalves; de Souza Ferreira, Luís Carlos; Benedetti, Celso E.; Blundell, Tom L.; Balan, Andrea

    2013-01-01

    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. PMID:24282519

  17. 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.

  18. Desenvolvimento de Planococcus citri (Risso, 1813 (Hemiptera: Pseudococcidae em cafeeiros = Development of Planococcus citri (Risso, 1813 (Hemiptera: Pseudococcidae in coffee trees

    Directory of Open Access Journals (Sweden)

    Lenira Viana Costa Santa-Cecília

    2009-01-01

    Full Text Available A cochonilha Planococcus citri (Risso, 1813 suga a seiva dos botões florais e frutos do cafeeiro, atacando as rosetas desde a floração até a colheita. Embora seja relatada há alguns anos na cafeicultura, são escassas as informações sobre o desenvolvimento dessacochonilha em cafeeiros. Assim, o presente trabalho teve por objetivos avaliar alguns aspectos biológicos da fase ninfal de P. citri em plantas de café. Ovos dessa cochonilha foram retirados de uma criação em laboratório, isolados em placas de Petri contendo discos foliares de Coffea arabica L., das cultivares Acaiá Cerrado, Mundo Novo e Catuaí Vermelho e de C. canephora Pierre & Froenher, cultivar Apoatã. As placas foram mantidas em câmara climatizada a 25 ± 1°C, 70 ± 10% de umidade relativa e 12h de fotofase. Constatou-se que a cultivar Catuaí Vermelho foi a que proporcionou maior duração do período ninfal das fêmeas, porém, não foram constatadas diferenças na mortalidade. Essa cochonilha se desenvolveu satisfatoriamente em todas as cultivares de café estudadas e os resultados não mostraram diferenças claras de susceptibilidade.The citrus mealybug Planococcus citri (Risso, 1813 feeds mainly on flowers and fruits of coffee plants from blooming until harvest. Little is known about its development in coffee although its occurrence is already known for several years. This work aims to study the nymph development of this mealybug in coffee plants.Eggs were isolated from a stock culture kept in laboratory and placed inside Petri dishes containing leaf sections in agar. The tested cultivars were Acaia Cerrado, Mundo Novo, Catuai Vermelho (Coffea arabica and Apoatã (C. canephora. Insects were kept in a climatizedchamber at 25ºC, 70 ± 10% humidity and 12-hour photophase. The longest development period in females was obtained in cultivar Catuai Vermelho. No differences in mortality were found among cultivars. Mealybugs developed in all cultivars and the results

  19. The abundant extrachromosomal DNA content of the Spiroplasma citri GII3-3X genome

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    Carrère Sébastien

    2008-04-01

    Full Text Available Abstract Background Spiroplama citri, the causal agent of citrus stubborn disease, is a bacterium of the class Mollicutes and is transmitted by phloem-feeding leafhopper vectors. In order to characterize candidate genes potentially involved in spiroplasma transmission and pathogenicity, the genome of S. citri strain GII3-3X is currently being deciphered. Results Assembling 20,000 sequencing reads generated seven circular contigs, none of which fit the 1.8 Mb chromosome map or carried chromosomal markers. These contigs correspond to seven plasmids: pSci1 to pSci6, with sizes ranging from 12.9 to 35.3 kbp and pSciA of 7.8 kbp. Plasmids pSci were detected as multiple copies in strain GII3-3X. Plasmid copy numbers of pSci1-6, as deduced from sequencing coverage, were estimated at 10 to 14 copies per spiroplasma cell, representing 1.6 Mb of extrachromosomal DNA. Genes encoding proteins of the TrsE-TraE, Mob, TraD-TraG, and Soj-ParA protein families were predicted in most of the pSci sequences, in addition to members of 14 protein families of unknown function. Plasmid pSci6 encodes protein P32, a marker of insect transmissibility. Plasmids pSci1-5 code for eight different S. citri adhesion-related proteins (ScARPs that are homologous to the previously described protein P89 and the S. kunkelii SkARP1. Conserved signal peptides and C-terminal transmembrane alpha helices were predicted in all ScARPs. The predicted surface-exposed N-terminal region possesses the following elements: (i 6 to 8 repeats of 39 to 42 amino acids each (sarpin repeats, (ii a central conserved region of 330 amino acids followed by (iii a more variable domain of about 110 amino acids. The C-terminus, predicted to be cytoplasmic, consists of a 27 amino acid stretch enriched in arginine and lysine (KR and an optional 23 amino acid stretch enriched in lysine, aspartate and glutamate (KDE. Plasmids pSci mainly present a linear increase of cumulative GC skew except in regions presenting

  20. 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

  1. 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.

  2. Influence of carrot psyllid (Trioza apicalis) feeding or exogenous limonene or methyl jasmonate treatment on composition of carrot (Daucus carota) leaf essential oil and headspace volatiles.

    Science.gov (United States)

    Nissinen, Anne; Ibrahim, Mohamed; Kainulainen, Pirjo; Tiilikkala, Kari; Holopainen, Jarmo K

    2005-11-02

    The effect of carrot psyllid (Trioza apicalis Förster) feeding and limonene and methyl jasmonate (MeJA) treatments on the essential oil composition and headspace volatiles of carrot (Daucus carota ssp. sativus), cvs. Parano and Splendid, leaves was studied. Carrot psyllid feeding significantly increased the concentrations of sabinene, beta-pinene, and limonene, whereas limonene treatment increased the concentration of (Z)-beta-ocimene in the leaves of both cultivars. The limonene treatment significantly increased the concentration of total phenolics in the leaves of both cultivars, and MeJA treatment increased phenolic concentration in the leaves of Parano. Exogenous limonene spray did not decrease the number of carrot psyllid eggs laid either 2 or 24 h after treatment. The results suggest that carrot psyllid feeding induces changes in the endogenous monoterpene pool in the carrot leaves. Limonene and MeJA treatments affect some induced defenses of the carrot, but the exogenous limonene spray is not an effective oviposition deterrent against carrot psyllid.

  3. 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.

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

    Science.gov (United States)

    Shen, Xiao-Min; Liao, Chong-Yu; Lu, Xue-Ping; Wang, Zhe; Wang, Jin-Jun; Dou, Wei

    2016-08-19

    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.

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

    Science.gov (United States)

    Piazza, Ainelén; Zimaro, Tamara; Garavaglia, Betiana S; Ficarra, Florencia A; Thomas, Ludivine; Marondedze, Claudius; Feil, Regina; Lunn, John E; Gehring, Chris; Ottado, Jorgelina; Gottig, Natalia

    2015-05-01

    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.

  6. 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.

  7. Responsiveness of different citrus genotypes to the Xanthomonas citri ssp. citri-derived pathogen-associated molecular pattern (PAMP) flg22 correlates with resistance to citrus canker.

    Science.gov (United States)

    Shi, Qingchun; Febres, Vicente J; Jones, Jeffrey B; Moore, Gloria A

    2015-06-01

    The bacterial agent of citrus canker disease (Xanthomonas citri ssp. citri, Xcc) has caused tremendous economic losses to the citrus industry around the world. Pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) is important to plant immunity. In this study, we compared the defence responses of citrus canker-resistant and citrus canker-susceptible genotypes to the Xcc-derived PAMP flg22 (Xflg22) by analysing the expression of 20 citrus defence-associated genes. We showed that, in the most resistant genotype, 'Nagami' kumquat, there was significant induction of several defence genes (EDS1, NDR1, PBS1, RAR1, SGT1, PAL1, NPR2 and NPR3) as early as 6 h and up to 72 h after Xflg22 treatment. At the other end of the spectrum, highly susceptible 'Duncan' grapefruit showed no induction of the same defence genes, even 120 h after treatment. Citrus genotypes with partial levels of resistance showed intermediate levels of transcriptional reprogramming that correlated with their resistance level. Xflg22 also triggered a rapid oxidative burst in all genotypes which was higher and accompanied by the induction of PTI marker genes (WRKY22 and GST1) only in the more resistant genotypes. Pretreatment with Xflg22 prior to Xcc inoculation inhibited bacterial growth in kumquat, but not in grapefruit. A flagellin-deficient Xcc strain (XccΔfliC) showed greater growth increase relative to wild-type Xcc in kumquat than in grapefruit. Taken together, our results indicate that Xflg22 initiates strong PTI in canker-resistant genotypes, but not in susceptible ones, and that a robust induction of PTI is an important component of citrus resistance to canker.

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

    Directory of Open Access Journals (Sweden)

    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

  9. Identification of differentially expressed genes of Xanthomonas axonopodis pv. citri by representational difference analysis of cDNA

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    Angela Mehta

    2005-03-01

    Full Text Available Xanthomonas axonopodis pv. citri is a phytopathogenic bacterium responsible for citrus canker, a serious disease which causes severe losses in citriculture around the world. In this study we report the differential expression of X. axonopodis pv. citri in response to specific treatments by using Representational Difference Analysis of cDNA (cDNA RDA. cDNAs from X. axonopodis pv. citri cultured in the presence of leaf extract of the host plant (Citrus sinensis, in vivo, as well as in the complex medium were hybridized against cDNA of the bacterium grown in the minimal medium. Sequencing of the difference products obtained after the second and third hybridizations revealed a total of 37 distinct genes identified by homology searches in the genome of X. axonopodis pv. citri. These genes were distributed in different functional categories, including genes that encode hypothetical proteins, genes involved in metabolism, cellular processes and pathogenicity, and mobile genetic elements. Most of these genes are likely related to growth and/or acquisition of nutrients in specific treatments whereas others might be important for the bacterium pathogenicity.

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

    Energy Technology Data Exchange (ETDEWEB)

    Faria, J.N.; Balan, A. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil); Paes Leme, A.F. [Laboratorio Nacional de Luz Sincrotron (LNLS), Campinas, SP (Brazil)

    2012-07-01

    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)

  11. Species limits in Diaporthe: molecular re-assessment of D. citri, D. cytosporella, D. foeniculina and D. rudis

    NARCIS (Netherlands)

    Udayanga, D.; Castlebury, L.A.; Rossman, A.Y.; Hyde, K.D.

    2014-01-01

    Species of Diaporthe are important plant pathogens of a wide range of hosts worldwide. In the present study the species causing melanose and stem end rot diseases of Citrus spp. are revised. Three species of Diaporthe occurring on Citrus are characterised, including D. citri, D. cytosporella and D.

  12. Identification and Characterization of Seven Glutathione S-Transferase Genes from Citrus Red Mite, Panonychus citri (McGregor

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    Chong-Yu Liao

    2013-12-01

    Full Text Available The citrus red mite, Panonychus citri (McGregor, is a global citrus pest, and has developed severe resistance to several types of acaricides. However, the molecular mechanisms of resistance in this mite remain unknown. In this study, seven full-length cDNAs encoding glutathione S-transferases (GSTs genes were identified and characterized in P. citri. The effects of pyridaben and fenpropathrin exposure on the expression of these genes were also investigated. Phylogenetic analysis revealed that the seven GSTs genes in P. citri cloned in this study belong to three different cytosolic classes, including four in mu, two in delta and one in zeta. Among these seven GSTs genes, the relative expression level of PcGSTm1 was significantly higher in adult than in the other life stages (egg, larvae and nymph. Compared with the control, the mRNA levels of the seven GST genes did not change significantly following exposure to pyridaben at LC10. However, RT-qPCR results showed that, when exposed to LC10 of fenpropathrin, six GSTs gene (PcGSTm1, PcGSTm3, PcGSTm4, PcGSTd1, PcGSTd2 and PcGSTz1 transcripts increased in a time-dependent manner. This is the first insight into the molecular characteristics of GSTs gene cDNAs in P. citri. The elevated GSTs gene transcripts following exposure to fenpropathrin might be one of the mechanisms involved in detoxification of this acaricide.

  13. Complete genome sequence of Spiroplasma citri strain R8-A2T, causal agent of stubborn disease in Citrus spp.

    Science.gov (United States)

    Spiroplasma citri is the causal agent of stubborn disease in Citrus spp., as well as the cause of diseases in numerous other plant genera. Here we report the nucleotide sequence of the 1,599,709 bp circular chromosome and two plasmids of strain R8-A2T. This information will facilitate comparative ...

  14. 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.

  15. 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)

  16. Development and reproduction of Panonychus citri (Prostigmata: Tetranychidae) on different species and varieties of citrus plants.

    Science.gov (United States)

    Zanardi, Odimar Zanuzo; Bordini, Gabriela Pavan; Franco, Aline Aparecida; de Morais, Matheus Rovere; Yamamoto, Pedro Takao

    2015-12-01

    The species and varieties of citrus plants that are currently grown can favor the population growth of the citrus red mite Panonychus citri (McGregor) (Prostigmata: Tetranychidae) and alter the pest management programs in citrus groves. In this study we evaluated, in the laboratory, the development and reproduction of P. citri and estimated its life table parameters when reared on four varieties of Citrus sinensis (L.) Osbeck (Valencia, Pera, Natal, and Hamlin), one variety of Citrus reticulata Blanco (Ponkan) and one variety of Citrus limon (L.) Burm. (Sicilian). The incubation period and egg viability were not affected by the host plant. However, the development and survival of the immature stage were significantly lower on Hamlin orange than on Valencia, Pera and Natal oranges, Ponkan mandarin and Sicilian lemon. The fecundity and oviposition period of females were lower on Hamlin orange than on the other hosts. Mites reared on Valencia orange and Sicilian lemon had a higher net reproductive rate (R 0 ), intrinsic growth rate (r) and finite rate of increase (λ), and a shorter interval between generations (T) than on Pera, Natal and Hamlin oranges and Ponkan mandarin. On the other hand, mites reared on Hamlin orange had the lowest R 0 , r and λ and the highest T among the hosts. Based on the results obtained we recommend that for Valencia orange and Sicilian lemon, the mite monitoring programs should be more intense to detect the initial infestation of pest, avoiding the damage in plants and the increase in production costs.

  17. A novel two-component response regulator links rpf with biofilm formation and virulence of Xanthomonas axonopodis pv. citri.

    Directory of Open Access Journals (Sweden)

    Tzu-Pi Huang

    Full Text Available Citrus bacterial canker caused by Xanthomonas axonopodis pv. citri is a serious disease that impacts citrus production worldwide, and X. axonopodis pv. citri is listed as a quarantine pest in certain countries. Biofilm formation is important for the successful development of a pathogenic relationship between various bacteria and their host(s. To understand the mechanisms of biofilm formation by X. axonopodis pv. citri strain XW19, the strain was subjected to transposon mutagenesis. One mutant with a mutation in a two-component response regulator gene that was deficient in biofilm formation on a polystyrene microplate was selected for further study. The protein was designated as BfdR for biofilm formation defective regulator. BfdR from strain XW19 shares 100% amino acid sequence identity with XAC1284 of X. axonopodis pv. citri strain 306 and 30-100% identity with two-component response regulators in various pathogens and environmental microorganisms. The bfdR mutant strain exhibited significantly decreased biofilm formation on the leaf surfaces of Mexican lime compared with the wild type strain. The bfdR mutant was also compromised in its ability to cause canker lesions. The wild-type phenotype was restored by providing pbfdR in trans in the bfdR mutant. Our data indicated that BfdR did not regulate the production of virulence-related extracellular enzymes including amylase, lipase, protease, and lecithinase or the expression of hrpG, rfbC, and katE; however, BfdR controlled the expression of rpfF in XVM2 medium, which mimics cytoplasmic fluids in planta. In conclusion, biofilm formation on leaf surfaces of citrus is important for canker development in X. axonopodis pv. citri XW19. The process is controlled by the two-component response regulator BfdR via regulation of rpfF, which is required for the biosynthesis of a diffusible signal factor.

  18. A LOV protein modulates the physiological attributes of Xanthomonas axonopodis pv. citri relevant for host plant colonization.

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    Ivana Kraiselburd

    Full Text Available Recent studies have demonstrated that an appropriate light environment is required for the establishment of efficient vegetal resistance responses in several plant-pathogen interactions. The photoreceptors implicated in such responses are mainly those belonging to the phytochrome family. Data obtained from bacterial genome sequences revealed the presence of photosensory proteins of the BLUF (Blue Light sensing Using FAD, LOV (Light, Oxygen, Voltage and phytochrome families with no known functions. Xanthomonas axonopodis pv. citri is a Gram-negative bacterium responsible for citrus canker. The in silico analysis of the X. axonopodis pv. citri genome sequence revealed the presence of a gene encoding a putative LOV photoreceptor, in addition to two genes encoding BLUF proteins. This suggests that blue light sensing could play a role in X. axonopodis pv. citri physiology. We obtained the recombinant Xac-LOV protein by expression in Escherichia coli and performed a spectroscopic analysis of the purified protein, which demonstrated that it has a canonical LOV photochemistry. We also constructed a mutant strain of X. axonopodis pv. citri lacking the LOV protein and found that the loss of this protein altered bacterial motility, exopolysaccharide production and biofilm formation. Moreover, we observed that the adhesion of the mutant strain to abiotic and biotic surfaces was significantly diminished compared to the wild-type. Finally, inoculation of orange (Citrus sinensis leaves with the mutant strain of X. axonopodis pv. citri resulted in marked differences in the development of symptoms in plant tissues relative to the wild-type, suggesting a role for the Xac-LOV protein in the pathogenic process. Altogether, these results suggest the novel involvement of a photosensory system in the regulation of physiological attributes of a phytopathogenic bacterium. A functional blue light receptor in Xanthomonas spp. has been described for the first time, showing

  19. Seasonal variation of natural mortality factors of the guava psyllid Triozoida limbata.

    Science.gov (United States)

    Semeão, A A; Martins, J C; Picanço, M C; Chediak, M; da Silva, E M; Silva, G A

    2012-12-01

    It is important to understand how components of the agroecosystem interfere with the attack of a pest species and their seasonality in order to use these components in IPM programs. This study focused on the evaluation of the seasonality of natural control factors associated with the guava psyllid Triozoida limbata (Enderlein) in Brazil. Life-table data were collected from an experimental guava orchard during four periods that roughly represented four seasons. Natural mortality was monitored daily through the immature stages, and the relative importance of each natural mortality factor and its seasonality was determined. Significant statistical differences were observed in the mortality during the four periods (P analysis (PCA) showed some trends in the relationship of natural control agents and weather conditions. For example, the occurrence of predatory wasps was positively correlated with temperature and occurrence of winds; the occurrence of syrphids and Psyllaephagus sp. were negatively correlated with temperature and winds; and the occurrence of other generalist predators were negatively correlated with the occurrence of rainfall and photoperiod. The results showed the importance of natural mortality factors for the management of T. limbata and their changes through the different seasons which should be considered when implementing IPM programs in guava orchards.

  20. Signalering van citruswolluis (Planococcus citri) in de teelt van diverse potplanten : onderzoek binnen project 41203147 "Verbetering biologische bestrijding van wolluis in diverse potplanten”

    NARCIS (Netherlands)

    Boertjes, B.C.; Bruin, de J.

    2003-01-01

    In 2002 en 2003 is door PPO Glastuinbouw het project “Verbetering biologische bestrijding van wolluis in diverse potplanten” (project 41203147) uitgevoerd. Binnen dit project werd onder meer onderzoek gedaan naar methoden om citruswolluis (Planococcus citri) te signaleren.

  1. KatG, the Bifunctional Catalase of Xanthomonas citri subsp. citri, Responds to Hydrogen Peroxide and Contributes to Epiphytic Survival on Citrus Leaves.

    Directory of Open Access Journals (Sweden)

    María Laura Tondo

    Full Text Available Xanthomonas citri subsp. citri (Xcc is the bacterium responsible for citrus canker. This bacterium is exposed to reactive oxygen species (ROS at different points during its life cycle, including those normally produced by aerobic respiration or upon exposition to ultraviolet (UV radiation. Moreover, ROS are key components of the host immune response. Among enzymatic ROS-detoxifying mechanisms, catalases eliminate H2O2, avoiding the potential damage caused by this specie. Xcc genome includes four catalase genes. In this work, we studied the physiological role of KatG, the only bifunctional catalase of Xcc, through the construction and characterization of a modified strain (XcckatG, carrying an insertional mutation in the katG gene. First, we evaluated the involvement of KatG in the bacterial adaptive response to H2O2. XcckatG cultures exhibited lower catalase activity than those of the wild-type strain, and this activity was not induced upon treatment with sub-lethal doses of H2O2. Moreover, the KatG-deficient mutant exhibited decreased tolerance to H2O2 toxicity compared to wild-type cells and accumulated high intracellular levels of peroxides upon exposure to sub-lethal concentrations of H2O2. To further study the role of KatG in Xcc physiology, we evaluated bacterial survival upon exposure to UV-A or UV-B radiation. In both conditions, XcckatG showed a high mortality in comparison to Xcc wild-type. Finally, we studied the development of bacterial biofilms. While structured biofilms were observed for the Xcc wild-type, the development of these structures was impaired for XcckatG. Based on these results, we demonstrated that KatG is responsible for Xcc adaptive response to H2O2 and a key component of the bacterial response to oxidative stress. Moreover, this enzyme plays an important role during Xcc epiphytic survival, being essential for biofilm formation and UV resistance.

  2. The gpsX gene encoding a glycosyltransferase is important for polysaccharide production and required for full virulence in Xanthomonas citri subsp. citri

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    Li Jinyun

    2012-03-01

    Full Text Available Abstract Background The Gram-negative bacterium Xanthomonas citri subsp. citri (Xac causes citrus canker, one of the most destructive diseases of citrus worldwide. In our previous work, a transposon mutant of Xac strain 306 with an insertion in the XAC3110 locus was isolated in a screening that aimed at identifying genes related to biofilm formation. The XAC3110 locus was named as bdp24 for biofilm-defective phenotype and the mutant was observed to be affected in extracellular polysaccharide (EPS and lipopolysaccharide (LPS biosynthesis and cell motility. In this study, we further characterized the bdp24 (XAC3110 gene (designated as gpsX using genetic complementation assays and expanded the knowledge about the function of the gpsX gene in Xac pathogenesis by investigating the roles of gpsX in EPS and LPS production, cell motility, biofilm formation on host leaves, stress tolerance, growth in planta, and host virulence of the citrus canker bacterium. Results The gpsX gene encodes a putative glycosyltransferase, which is highly conserved in the sequenced strains of Xanthomonas. Mutation of gpsX resulted in a significant reduction of the amount of EPS and loss of two LPS bands visualized on sodium dodecylsulphate- polyacrylamide gels. Biofilm assays revealed that the gpsX mutation affected biofilm formation by Xac on abiotic and biotic surfaces. The gpsX mutant showed delayed bacterial growth and caused reduced development of disease symptoms in susceptible citrus leaves. The gpsX mutant was more sensitive than the wild-type strain to various stresses, including the H2O2 oxidative stress. The mutant also showed attenuated ability in cell motility but not in flagellar formation. Quantitative reverse transcription-PCR assays indicated that mutation of gpsX did not affect the expression of virulence genes such as pthA in Xac strain 306. The affected phenotypes of the gpsX mutant could be complemented to wild-type levels by the intact gpsX gene

  3. KatG, the Bifunctional Catalase of Xanthomonas citri subsp. citri, Responds to Hydrogen Peroxide and Contributes to Epiphytic Survival on Citrus Leaves.

    Science.gov (United States)

    Tondo, María Laura; Delprato, María Laura; Kraiselburd, Ivana; Fernández Zenoff, María Verónica; Farías, María Eugenia; Orellano, Elena G

    2016-01-01

    Xanthomonas citri subsp. citri (Xcc) is the bacterium responsible for citrus canker. This bacterium is exposed to reactive oxygen species (ROS) at different points during its life cycle, including those normally produced by aerobic respiration or upon exposition to ultraviolet (UV) radiation. Moreover, ROS are key components of the host immune response. Among enzymatic ROS-detoxifying mechanisms, catalases eliminate H2O2, avoiding the potential damage caused by this specie. Xcc genome includes four catalase genes. In this work, we studied the physiological role of KatG, the only bifunctional catalase of Xcc, through the construction and characterization of a modified strain (XcckatG), carrying an insertional mutation in the katG gene. First, we evaluated the involvement of KatG in the bacterial adaptive response to H2O2. XcckatG cultures exhibited lower catalase activity than those of the wild-type strain, and this activity was not induced upon treatment with sub-lethal doses of H2O2. Moreover, the KatG-deficient mutant exhibited decreased tolerance to H2O2 toxicity compared to wild-type cells and accumulated high intracellular levels of peroxides upon exposure to sub-lethal concentrations of H2O2. To further study the role of KatG in Xcc physiology, we evaluated bacterial survival upon exposure to UV-A or UV-B radiation. In both conditions, XcckatG showed a high mortality in comparison to Xcc wild-type. Finally, we studied the development of bacterial biofilms. While structured biofilms were observed for the Xcc wild-type, the development of these structures was impaired for XcckatG. Based on these results, we demonstrated that KatG is responsible for Xcc adaptive response to H2O2 and a key component of the bacterial response to oxidative stress. Moreover, this enzyme plays an important role during Xcc epiphytic survival, being essential for biofilm formation and UV resistance.

  4. KatG, the Bifunctional Catalase of Xanthomonas citri subsp. citri, Responds to Hydrogen Peroxide and Contributes to Epiphytic Survival on Citrus Leaves

    Science.gov (United States)

    Tondo, María Laura; Delprato, María Laura; Kraiselburd, Ivana; Fernández Zenoff, María Verónica; Farías, María Eugenia; Orellano, Elena G.

    2016-01-01

    Xanthomonas citri subsp. citri (Xcc) is the bacterium responsible for citrus canker. This bacterium is exposed to reactive oxygen species (ROS) at different points during its life cycle, including those normally produced by aerobic respiration or upon exposition to ultraviolet (UV) radiation. Moreover, ROS are key components of the host immune response. Among enzymatic ROS-detoxifying mechanisms, catalases eliminate H2O2, avoiding the potential damage caused by this specie. Xcc genome includes four catalase genes. In this work, we studied the physiological role of KatG, the only bifunctional catalase of Xcc, through the construction and characterization of a modified strain (XcckatG), carrying an insertional mutation in the katG gene. First, we evaluated the involvement of KatG in the bacterial adaptive response to H2O2. XcckatG cultures exhibited lower catalase activity than those of the wild-type strain, and this activity was not induced upon treatment with sub-lethal doses of H2O2. Moreover, the KatG-deficient mutant exhibited decreased tolerance to H2O2 toxicity compared to wild-type cells and accumulated high intracellular levels of peroxides upon exposure to sub-lethal concentrations of H2O2. To further study the role of KatG in Xcc physiology, we evaluated bacterial survival upon exposure to UV-A or UV-B radiation. In both conditions, XcckatG showed a high mortality in comparison to Xcc wild-type. Finally, we studied the development of bacterial biofilms. While structured biofilms were observed for the Xcc wild-type, the development of these structures was impaired for XcckatG. Based on these results, we demonstrated that KatG is responsible for Xcc adaptive response to H2O2 and a key component of the bacterial response to oxidative stress. Moreover, this enzyme plays an important role during Xcc epiphytic survival, being essential for biofilm formation and UV resistance. PMID:26990197

  5. 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)

    Asian citrus psyllid (ACP) feeding behaviors play a significant role in the transmission of the phloem-limited Candidatus Liberibacter asiaticus (CLas) bacteria that cause citrus greening disease. Sustained phloem ingestion by ACP on CLas infected plants is very important in pathogen acquisition and...

  6. First Report of "Candidatus Liberibacter psyllaurous" (synonym "Ca. L. solanacearum") Associated with 'Tomato Vein-Greening' and 'Tomato psyllid yellows' Diseases in Commercial Greenhouses in Arizona

    Science.gov (United States)

    During 2006-2007, tomato plants in two independent, commercial greenhouses in Arizona were infested with potato psyllid Paratrioza cockerelli. Over 60% and ~20% of plants in GH-1 and GH-2, respectively, exhibited leaf curling, stunting, and shortened internodes, and GH-1 plants also showed vein-gree...

  7. New geographic records for the Nearctic psyllid Bactericera maculipennis (Crawford) with biological notes and descriptions of the egg and fifth-instar nymph (Hemiptera: Psylloidea: Triozidae)

    Science.gov (United States)

    The Nearctic psyllid Bactericera maculipennis (Crawford) is one of only four known species of Psylloidea worldwide whose primary hosts are species of Convolvulaceae. Taxonomic syntheses of the Nearctic Psylloidea list B. maculipennis only from California and Utah. Our surveys of field bindweed, Co...

  8. Bionomics of Asian citrus psyllid associated with orange jasmine hedges in Florida, with special reference to biological control by Tamarixia radiata

    Science.gov (United States)

    The Asian citrus psyllid (ACP) is an important invasive citrus pest because it vectors a bacterium responsible for a devastating disease of citrus known as huanglongbing. Orange jasmine (Murraya paniculata) is a favored alternate ACP host plant and is widely grown as an ornamental plant in urban ar...

  9. Overexpression of citrus polygalacturonase-inhibiting protein in citrus black rot pathogen Alternaria citri.

    Science.gov (United States)

    Katoh, Hiroshi; Nalumpang, Sarunya; Yamamoto, Hiroyuki; Akimitsu, Kazuya

    2007-05-01

    The rough lemon (Citrus jambhiri) gene encoding polygalacturonase-inhibiting protein (RlemPGIPA) was overexpressed in the pathogenic fungus Alternaria citri. The overexpression mutant AcOPI6 retained the ability to utilize pectin as a sole carbon source, and the overexpression of polygalacturonase-inhibiting protein did not have any effect on the growth of AcOPI6 in potato dextrose and pectin medium. The pathogenicity of AcOPI6 to cause a black rot symptom in citrus fruits was also unchanged. Polygalacturonase-inhibiting protein was secreted together with endopolygalacturonase into culture filtrates of AcOPI6, and oligogalacturonides were digested from polygalacturonic acid by both proteins in the culture filtrates. The reaction mixture containing oligogalacturonides possessed activity for induction of defense-related gene, RlemLOX, in rough lemon leaves.

  10. Chemotactic signal transduction and phosphate metabolism as adaptive strategies during citrus canker induction by Xanthomonas citri.

    Science.gov (United States)

    Moreira, Leandro Marcio; Facincani, Agda Paula; Ferreira, Cristiano Barbalho; Ferreira, Rafael Marine; Ferro, Maria Inês Tiraboshi; Gozzo, Fabio Cesar; de Oliveira, Julio Cezar Franco; Ferro, Jesus Aparecido; Soares, Márcia Regina

    2015-03-01

    The genome of Xanthomonas citri subsp. Citri strain 306 pathotype A (Xac) was completely sequenced more than 10 years; to date, few studies involving functional genomics Xac and its host compatible have been developed, specially related to adaptive events that allow the survival of Xac within the plant. Proteomic analysis of Xac showed that the processes of chemotactic signal transduction and phosphate metabolism are key adaptive strategies during the interaction of a pathogenic bacterium with its plant host. The results also indicate the importance of a group of proteins that may not be directly related to the classical virulence factors, but that are likely fundamental to the success of the initial stages of the infection, such as methyl-accepting chemotaxis protein (Mcp) and phosphate specific transport (Pst). Furthermore, the analysis of the mutant of the gene pstB which codifies to an ABC phosphate transporter subunit revealed a complete absence of citrus canker symptoms when inoculated in compatible hosts. We also conducted an in silico analysis which established the possible network of genes regulated by two-component systems PhoPQ and PhoBR (related to phosphate metabolism), and possible transcriptional factor binding site (TFBS) motifs of regulatory proteins PhoB and PhoP, detaching high degree of conservation of PhoB TFBS in 84 genes of Xac genome. This is the first time that chemotaxis signal transduction and phosphate metabolism were therefore indicated to be fundamental to the process of colonization of plant tissue during the induction of disease associated with Xanthomonas genus bacteria.

  11. Digital Gene Expression Analysis of Ponkan Mandarin (Citrus reticulata Blanco) in Response to Asia Citrus Psyllid-Vectored Huanglongbing Infection.

    Science.gov (United States)

    Zhong, Yun; Cheng, Chunzhen; Jiang, Bo; Jiang, Nonghui; Zhang, Yongyan; Hu, Minlun; Zhong, Guangyan

    2016-07-02

    Citrus Huanglongbing (HLB), the most destructive citrus disease, can be transmitted by psyllids and diseased budwoods. Although the final symptoms of the two main HLB transmission ways were similar and hard to distinguish, the host responses might be different. In this study, the global gene changes in leaves of ponkan (Citrus reticulata) mandarin trees following psyllid-transmission of HLB were analyzed at the early symptomatic stage (13 weeks post inoculation, wpi) and late symptomatic stage (26 wpi) using digital gene expression (DGE) profiling. At 13 wpi, 2452 genes were downregulated while only 604 genes were upregulated in HLB infected ponkan leaves but no pathway enrichment was identified. Gene function analysis showed impairment in defense at the early stage of infection. At late stage of 26 wpi, however, differentially expressed genes (DEGs) involved in carbohydrate metabolism, plant defense, hormone signaling, secondary metabolism, transcription regulation were overwhelmingly upregulated, indicating that the defense reactions were eventually activated. The results indicated that HLB bacterial infection significantly influenced ponkan gene expression, and a delayed response of the host to the fast growing bacteria might be responsible for its failure in fighting against the bacteria.

  12. Digital Gene Expression Analysis of Ponkan Mandarin (Citrus reticulata Blanco in Response to Asia Citrus Psyllid-Vectored Huanglongbing Infection

    Directory of Open Access Journals (Sweden)

    Yun Zhong

    2016-07-01

    Full Text Available Citrus Huanglongbing (HLB, the most destructive citrus disease, can be transmitted by psyllids and diseased budwoods. Although the final symptoms of the two main HLB transmission ways were similar and hard to distinguish, the host responses might be different. In this study, the global gene changes in leaves of ponkan (Citrus reticulata mandarin trees following psyllid-transmission of HLB were analyzed at the early symptomatic stage (13 weeks post inoculation, wpi and late symptomatic stage (26 wpi using digital gene expression (DGE profiling. At 13 wpi, 2452 genes were downregulated while only 604 genes were upregulated in HLB infected ponkan leaves but no pathway enrichment was identified. Gene function analysis showed impairment in defense at the early stage of infection. At late stage of 26 wpi, however, differentially expressed genes (DEGs involved in carbohydrate metabolism, plant defense, hormone signaling, secondary metabolism, transcription regulation were overwhelmingly upregulated, indicating that the defense reactions were eventually activated. The results indicated that HLB bacterial infection significantly influenced ponkan gene expression, and a delayed response of the host to the fast growing bacteria might be responsible for its failure in fighting against the bacteria.

  13. Molecular and physiological properties associated with zebra complex disease in potatoes and its relation with Candidatus Liberibacter contents in psyllid vectors.

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    Veria Y Alvarado

    Full Text Available Zebra complex (ZC disease on potatoes is associated with Candidatus Liberibacter solanacearum (CLs, an α-proteobacterium that resides in the plant phloem and is transmitted by the potato psyllid Bactericera cockerelli (Šulc. The name ZC originates from the brown striping in fried chips of infected tubers, but the whole plants also exhibit a variety of morphological features and symptoms for which the physiological or molecular basis are not understood. We determined that compared to healthy plants, stems of ZC-plants accumulate starch and more than three-fold total protein, including gene expression regulatory factors (e.g. cyclophilin and tuber storage proteins (e.g., patatins, indicating that ZC-affected stems are reprogrammed to exhibit tuber-like physiological properties. Furthermore, the total phenolic content in ZC potato stems was elevated two-fold, and amounts of polyphenol oxidase enzyme were also high, both serving to explain the ZC-hallmark rapid brown discoloration of air-exposed damaged tissue. Newly developed quantitative and/or conventional PCR demonstrated that the percentage of psyllids in laboratory colonies containing detectable levels of CLs and its titer could fluctuate over time with effects on colony prolificacy, but presumed reproduction-associated primary endosymbiont levels remained stable. Potato plants exposed in the laboratory to psyllid populations with relatively low-CLs content survived while exposure of plants to high-CLs psyllids rapidly culminated in a lethal collapse. In conclusion, we identified plant physiological biomarkers associated with the presence of ZC and/or CLs in the vegetative potato plant tissue and determined that the titer of CLs in the psyllid population directly affects the rate of disease development in plants.

  14. Crystal Structure of the FAD-Containing Ferredoxin-NADP+ Reductase from the Plant Pathogen Xanthomonas axonopodis pv. citri

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    María Laura Tondo

    2013-01-01

    Full Text Available We have solved the structure of ferredoxin-NADP(H reductase, FPR, from the plant pathogen Xanthomonas axonopodis pv. citri, responsible for citrus canker, at a resolution of 1.5 Å. This structure reveals differences in the mobility of specific loops when compared to other FPRs, probably unrelated to the hydride transfer process, which contributes to explaining the structural and functional divergence between the subclass I FPRs. Interactions of the C-terminus of the enzyme with the phosphoadenosine of the cofactor FAD limit its mobility, thus affecting the entrance of nicotinamide into the active site. This structure opens the possibility of rationally designing drugs against the X. axonopodis pv. citri phytopathogen.

  15. Influence of field attack by carrot psyllid (Trioza apicalis Förster) on sensory quality, antioxidant capacity and content of terpenes, falcarindiol and 6-methoxymellein of carrots (Daucus carota L.).

    Science.gov (United States)

    Seljåsen, Randi; Vogt, Gjermund; Olsen, Elisabeth; Lea, Per; Høgetveit, Lars Arne; Tajet, Torgeir; Meadow, Richard; Bengtsson, Gunnar B

    2013-03-20

    The effect of different degrees of attack by carrot psyllid (Trioza apicalis) on quality parameters of carrots was studied in field experiments for two years. Treatments were different degrees of physical insect protection by floating row cover. An increasing attack level of psyllids showed an enhancement effect on the antioxidant capacity (ORAC), content of falcarindiol, 6-methoxymellein, and terpenes, and scores for bitter taste, chemical flavor, terpene flavor, and toughness. Carrot psyllid attack decreased the yield, total sugar, fructose, glucose, and sensory attributes sweet taste, color hue, color strength, crispiness, and juiciness. Carrot plants at 8-10 weeks of age tolerated attack by psyllids at low levels (2% leaves with curling or discoloration).

  16. De novo characterization of the Dialeurodes citri transcriptome: mining genes involved in stress resistance and simple sequence repeats (SSRs) discovery.

    Science.gov (United States)

    Chen, E-H; Wei, D-D; Shen, G-M; Yuan, G-R; Bai, P-P; Wang, J-J

    2014-02-01

    The citrus whitefly, Dialeurodes citri (Ashmead), is one of the three economically important whitefly species that infest citrus plants around the world; however, limited genetic research has been focused on D. citri, partly because of lack of genomic resources. In this study, we performed de novo assembly of a transcriptome using Illumina paired-end sequencing technology (Illumina Inc., San Diego, CA, USA). In total, 36,766 unigenes with a mean length of 497 bp were identified. Of these unigenes, we identified 17,788 matched known proteins in the National Center for Biotechnology Information database, as determined by Blast search, with 5731, 4850 and 14,441 unigenes assigned to clusters of orthologous groups (COG), gene ontology (GO), and SwissProt, respectively. In total, 7507 unigenes were assigned to 308 known pathways. In-depth analysis of the data showed that 117 unigenes were identified as potentially involved in the detoxification of xenobiotics and 67 heat shock protein (Hsp) genes were associated with environmental stress. In addition, these enzymes were searched against the GO and COG database, and the results showed that the three major detoxification enzymes and Hsps were classified into 18 and 3, 6, and 8 annotations, respectively. In addition, 149 simple sequence repeats were detected. The results facilitate the investigation of molecular resistance mechanisms to insecticides and environmental stress, and contribute to molecular marker development. The findings greatly improve our genetic understanding of D. citri, and lay the foundation for future functional genomics studies on this species.

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

  18. Development of rapid, sensitive and non-radioactive tissue-blot diagnostic method for the detection of citrus greening.

    Science.gov (United States)

    Nageswara-Rao, Madhugiri; Miyata, Shin-Ichi; Ghosh, Dilip; Irey, Mike; Garnsey, Stephen M; Gowda, Siddarame

    2013-01-01

    Citrus huanglongbing (HLB or citrus greening) is one of the most devastating diseases of citrus worldwide. The disease is caused by Gram-negative, phloem-limited α-proteobacterium, 'Candidatus Liberibacter asiaticus', vectored by the psyllid, Diaphorina citri Kuwayama. Citrus plants infected by the HLB bacterium may not show visible symptoms sometimes for years following infection and non-uniform distribution within the tree makes the detection of the pathogen very difficult. Efficient management of HLB disease requires rapid and sensitive detection early in the infection followed by eradication of the source of pathogen and the vector. The polymerase chain reaction (PCR) based method is most commonly employed for screening the infected/suspected HLB plants and psyllids. This is time consuming, cumbersome and not practical for screening large number of samples in the field. To overcome this, we developed a simple, sensitive, non-radioactive, tissue-blot diagnostic method for early detection and screening of HLB disease. Digoxigenin labeled molecular probes specific to 'Ca. L. asiaticus' nucleotide sequences have been developed and used for the detection of the pathogen of the HLB disease. The copy number of the target genes was also assessed using real-time PCR experiments and the optimized real-time PCR protocol allowed positive 'Ca. L. asiaticus' detection in citrus samples infected with 'Ca. L. asiaticus' bacterium.

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

    Science.gov (United States)

    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.

  20. Polymerase chain reaction-restriction fragment length polymorphism method to distinguish three mealybug groups within the Planococcus citri-P. minor species complex (Hemiptera: Coccoidea: Pseudococcidae).

    Science.gov (United States)

    Rung, A; Miller, D R; Scheffer, S J

    2009-02-01

    The mealybug species Planococcus citri (Risso) and Planococcus minor (Maskell) (Hemiptera: Coccoidea: Pseudococcidae) have special significance to U.S. quarantine and U.S. agriculture. Commonly intercepted at U.S. ports-of-entry, they are difficult to identify based on morphological characters. This study presents a molecular method for distinguishing P. citri, P. minor, and a genetically distinct group that is morphologically identical to P. citri, from Hawaii. This method uses polymerase chain reaction (PCR) followed by restriction fragment polymorphism analysis (RFLP) using the restriction enzymes BspH1, BsmH1, and HpH1. The resulting band patterns can be visualized in a 2% agarose gel and are sufficient to differentiate between the three entities mentioned above. PCR-RFLP diagnostics can be used for all life stages and is cheaper and faster than DNA sequencing.

  1. The effect of Leptomastix dactylopii parasitism and venom injection on host Planococcus citri

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    D Battaglia

    2014-10-01

    Full Text Available One of the major alterations observed in mealybug Planococcus citri parasitized by Leptomastix dactylopii is a strong reduction of laid eggs, which is evident soon after parasitization. Venom injection in unparasitized hosts determines a drastic reduction of fecundity indicating that this female secretion injected at the oviposition plays a key-role in host regulation. In order to assess the impact of parasitism and venom injection on host reproductive tissues, ovaries were dissected at different time intervals after these treatments and observed by light and transmission electron microscopy. The developing eggs showed clear symptoms of degeneration, already half an hour after parasitization or venom injection. Heat and protease treatments of venom nearly suppressed its effects on host reproduction, indicating that proteins are likely responsible for the observed alterations. The electrophoretic profile of venom proteins covers a wide range of molecular masses between 15 to 200 kDa but five major bands having a molecular mass of about 27, 30, 40, 90 and 120 kDa respectively were more evident. Moreover, to establish any parasitoid preference in host selection, among the adult female mealybugs at different stages of maturation and a possible relation with fecundity reduction in the host, the parasitoid behavior was observed.

  2. 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.

  3. Identification of anti-asthmatic compounds in Pericarpium citri reticulatae and evaluation of their synergistic effects

    Institute of Scientific and Technical Information of China (English)

    Qian SHI; Ze LIU; Yang YANG; Peng GENG; Yuan-yuan ZHU; Qi ZHANG; Fang BAI; Gang BAI

    2009-01-01

    Aim: To investigate the anti-asthmatic mechanisms of the traditional Chinese medicine Pericarpium citri reticulatae (PCR).Methods: The alkaloid section (AS) of PCR was extracted using an ion exchange resin, separated, and purified into different fractions by semi-preparative HPLC. These fractions were screened for beta2-adrenergic receptor (β2AR) agonistic activity using rat β2AR-transfected CHO-CRE-EGFP cells. AS and its isolated components were characterized by ultra-performance liquid chromatography/quadrupole time-of-flight MS (UPLC/Q-Tof MS) and were evaluated for their spasmolytic and antitussive activities both in vitro and in vivo in a guinea pig model.Results: We demonstrated that the AS component responsible for activating β2AR signaling was synephrine. Both AS and synephrine showed significant spasmolytic effects on acetylcholine chloride (ACh)-induced contractions in isolated guinea pig trachea, and they protected against histamine-induced experimental asthma by prolonging the latent period. We further identified stachydrine as the antitussive component that could significantly reduce citric acid-induced coughing. The combination of these two bioactive compounds had a more potent spasmolytic activity in comparison with the single use of synephrine or stachydrine.Conclusion: We conclude that synephrine and stachydrine are the key components of AS that mediate asthma relief due to their synergism when used in combination.

  4. Genomic insights into the evolutionary origin of Xanthomonas axonopodis pv. citri and its ecological relatives.

    Science.gov (United States)

    Midha, Samriti; Patil, Prabhu B

    2014-10-01

    Xanthomonas axonopodis pv. citri (Xac) is the causal agent of citrus bacterial canker (CBC) and is a serious problem worldwide. Like CBC, several important diseases in other fruits, such as mango, pomegranate, and grape, are also caused by Xanthomonas pathovars that display remarkable specificity toward their hosts. While citrus and mango diseases were documented more than 100 years ago, the pomegranate and grape diseases have been known only since the 1950s and 1970s, respectively. Interestingly, diseases caused by all these pathovars were noted first in India. Our genome-based phylogenetic studies suggest that these diverse pathogens belong to a single species and these pathovars may be just a group of rapidly evolving strains. Furthermore, the recently reported pathovars, such as those infecting grape and pomegranate, form independent clonal lineages, while the citrus and mango pathovars that have been known for a long time form one clonal lineage. Such an understanding of their phylogenomic relationship has further allowed us to understand major and unique variations in the lineages that give rise to these pathovars. Whole-genome sequencing studies including ecological relatives from their putative country of origin has allowed us to understand the evolutionary history of Xac and other pathovars that infect fruits.

  5. Pathogenic Interactions Between Xanthomonas axonopodis pv. citri and Cultivars of Pummelo (Citrus grandis).

    Science.gov (United States)

    Shiotani, H; Ozaki, K; Tsuyumu, S

    2000-12-01

    ABSTRACT The aggressiveness of strains of Xanthomonas axonopodis pv. citri on seven Citrus species, including Citrus sinensis (navel orange), C. paradisi (grapefruit), C. unshiu (Satsuma mandarin), C. junos (Yuzu), C. aurantifolia ('Mexican' lime), C. tachibana (Tachibana), and C. grandis (pummelo: 'Otachibana', 'Banpeiyu', and 'Anseikan'), were assessed by comparing lesion expansion and growth in planta, using a prick inoculation method. The existence of two groups distinct in aggressiveness was demonstrated on the pummelo cultivars, whereas the remaining species tested were uniformly susceptible. The two groups of strains were distinct in lesion expansion and growth in planta; however, both caused canker lesions on the 'Otachibana' pummelo. The sensitivity of the bacterial strains to phages Cp1 and Cp2 was associated with differences in aggressiveness. Namely, all the strains sensitive to Cp2 but resistant to Cp1 were aggressive to 'Otachibana', whereas all the strains sensitive to Cp1 but resistant to Cp2 were weakly aggressive. When a repetitive sequence-based polymerase chain reaction amplification was carried out by enterobacterial repetitive intergeneric consensus (ERIC) sequences (ERIC1R and ERIC2) as the primers, these two groups were also distinguishable by the presence or absence of a 1.8-kb DNA fragment among otherwise identical fragments. The 1.8-kb fragment was amplified only from the strains aggressive to C. grandis.

  6. Remote Sensing of Low and Mid-Latitude Ionospheric Disturbances During Solar Minimum Using CITRIS and CERTO Measurements of TEC and Radio Scintillation

    Science.gov (United States)

    Siefring, C. L.; Bernhardt, P. A.

    2010-12-01

    Unique data on ionospheric plasma disturbances from the Naval Research Laboratory 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) three-frequency beacons transmitting at 150/400/1067 MHz and 2) the French global network of ground-based DORIS (Doppler Orbitography and Radiopositioning Integrated by Satellite) beacons transmitting at 401.25 and 2036.25 MHz. CITRIS was operated in a complementary fashion with the C/NOFS satellite during most of its first year of operations; C/NOFS carries CERTO beacon along with in-situ diagnostics. CITRIS and ground receivers can simultaneously measure TEC and scintillations on different paths using CERTO on C/NOFS. When C/NOFS is not in view, CITRIS makes measurements from DORIS beacons and other LEO satellites. Because of the orbits CITRIS will always make measurements at the same longitude within 48 min of C/NOFS. The ability to look at multiple paths is unique and useful for studying the spatial extent and time duration of disturbances. The combination of TEC and scintillation measurements provides information on a range of scale-sizes from >1 km to about 100 m. The joint data set on plasma structures at low-latitudes is a focus of our presentation, with the addition of comparisons to CITRIS data taken at mid-latitude. Several types of irregularities have been studied including Spread-F and the newly discovered dawn-side depletions. The data covers large portions of the Earth (including the Pacific, African and South American sectors) during an unusually quite portion of the most

  7. Characterization of bacteriophages Cp1 and Cp2, the strain-typing agents for Xanthomonas axonopodis pv. citri.

    Science.gov (United States)

    Ahmad, Abdelmonim Ali; Ogawa, Megumi; Kawasaki, Takeru; Fujie, Makoto; Yamada, Takashi

    2014-01-01

    The strains of Xanthomonas axonopodis pv. citri, the causative agent of citrus canker, are historically classified based on bacteriophage (phage) sensitivity. Nearly all X. axonopodis pv. citri strains isolated from different regions in Japan are lysed by either phage Cp1 or Cp2; Cp1-sensitive (Cp1(s)) strains have been observed to be resistant to Cp2 (Cp2(r)) and vice versa. In this study, genomic and molecular characterization was performed for the typing agents Cp1 and Cp2. Morphologically, Cp1 belongs to the Siphoviridae. Genomic analysis revealed that its genome comprises 43,870-bp double-stranded DNA (dsDNA), with 10-bp 3'-extruding cohesive ends, and contains 48 open reading frames. The genomic organization was similar to that of Xanthomonas phage phiL7, but it lacked a group I intron in the DNA polymerase gene. Cp2 resembles morphologically Escherichia coli T7-like phages of Podoviridae. The 42,963-bp linear dsDNA genome of Cp2 contained terminal repeats. The Cp2 genomic sequence has 40 open reading frames, many of which did not show detectable homologs in the current databases. By proteomic analysis, a gene cluster encoding structural proteins corresponding to the class III module of T7-like phages was identified on the Cp2 genome. Therefore, Cp1 and Cp2 were found to belong to completely different virus groups. In addition, we found that Cp1 and Cp2 use different molecules on the host cell surface as phage receptors and that host selection of X. axonopodis pv. citri strains by Cp1 and Cp2 is not determined at the initial stage by binding to receptors.

  8. 60Co-γ Irradiation Affects the Enzymatic Antioxidant System of the Citrus Red Mite Panonychus citri (Acari: Tetranychidae

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    Ke Zhang

    2014-05-01

    Full Text Available Radio-(60Co, which emits γ rays, has been used worldwide in pest control. The aim of the present study was to analyze the effect of effective-low-power 60Co-γ irradiation on the enzymatic antioxidant system of the citrus red mite Panonychus citri. One day old female adults were exposed to 0.4 kGy 60Co-γ irradiation and on the, 6th h, 1st day, 2nd day, and 5th day post treatment, the mites were euthanized for biochemical analysis. The activity of superoxide dismutase (SOD, catalase (CAT, peroxidase (POD, phenoloxidase (PO and acetylocholinesterase (AchE were investigated. POD and CAT activities, as well as SOD were higher in the irradiated mites. We found that exposure to 60Co-γ radiation resulted in increased activities of SOD, CAT, POD and decreased AchE activity. When the recovery time lasted till the 5th day, the activities of POD and PO were significantly lower than the control, whereas the SOD, CAT and AchE activities returned to control levels. Cells possess protein repair pathways to rescue oxidized proteins and restore their functions, but if these repair processes fail, oxidized proteins may become cytotoxic. Our results confirm the hypothesis that low dosages of 60Co-γ irradiation increase the level of oxidative stress in P. citri adults in a short time, causing meanwhile damage and sterility. The results of this study also indicate that stress caused by exposure to irradiation could inhibit the cholinergic system in P. citri.

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

    Energy Technology Data Exchange (ETDEWEB)

    Cary; R. Bruce (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.

  10. Structural-functional characterization and physiological significance of ferredoxin-NADP reductase from Xanthomonas axonopodis pv. citri.

    Science.gov (United States)

    Tondo, María Laura; Musumeci, Matías A; Delprato, María Laura; Ceccarelli, Eduardo A; Orellano, Elena G

    2011-01-01

    Xanthomonas axonopodis pv. citri is a phytopathogen bacterium that causes severe citrus canker disease. Similar to other phytopathogens, after infection by this bacterium, plants trigger a defense mechanism that produces reactive oxygen species. Ferredoxin-NADP(+) reductases (FNRs) are redox flavoenzymes that participate in several metabolic functions, including the response to reactive oxygen species. Xanthomonas axonopodis pv. citri has a gene (fpr) that encodes for a FNR (Xac-FNR) that belongs to the subclass I bacterial FNRs. The aim of this work was to search for the physiological role of this enzyme and to characterize its structural and functional properties. The functionality of Xac-FNR was tested by cross-complementation of a FNR knockout Escherichia coli strain, which exhibit high susceptibility to agents that produce an abnormal accumulation of (•)O(2)(-). Xac-FNR was able to substitute for the FNR in E. coli in its antioxidant role. The expression of fpr in X. axonopodis pv. citri was assessed using semiquantitative RT-PCR and Western blot analysis. A 2.2-fold induction was observed in the presence of the superoxide-generating agents methyl viologen and 2,3-dimethoxy-1,4-naphthoquinone. Structural and functional studies showed that Xac-FNR displayed different functional features from other subclass I bacterial FNRs. Our analyses suggest that these differences may be due to the unusual carboxy-terminal region. We propose a further classification of subclass I bacterial FNRs, which is useful to determine the nature of their ferredoxin redox partners. Using sequence analysis, we identified a ferredoxin (XAC1762) as a potential substrate of Xac-FNR. The purified ferredoxin protein displayed the typical broad UV-visible spectrum of [4Fe-4S] clusters and was able to function as substrate of Xac-FNR in the cytochrome c reductase activity. Our results suggest that Xac-FNR is involved in the oxidative stress response of Xanthomonas axonopodis pv. citri and

  11. Structural-functional characterization and physiological significance of ferredoxin-NADP reductase from Xanthomonas axonopodis pv. citri.

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    María Laura Tondo

    Full Text Available Xanthomonas axonopodis pv. citri is a phytopathogen bacterium that causes severe citrus canker disease. Similar to other phytopathogens, after infection by this bacterium, plants trigger a defense mechanism that produces reactive oxygen species. Ferredoxin-NADP(+ reductases (FNRs are redox flavoenzymes that participate in several metabolic functions, including the response to reactive oxygen species. Xanthomonas axonopodis pv. citri has a gene (fpr that encodes for a FNR (Xac-FNR that belongs to the subclass I bacterial FNRs. The aim of this work was to search for the physiological role of this enzyme and to characterize its structural and functional properties. The functionality of Xac-FNR was tested by cross-complementation of a FNR knockout Escherichia coli strain, which exhibit high susceptibility to agents that produce an abnormal accumulation of (•O(2(-. Xac-FNR was able to substitute for the FNR in E. coli in its antioxidant role. The expression of fpr in X. axonopodis pv. citri was assessed using semiquantitative RT-PCR and Western blot analysis. A 2.2-fold induction was observed in the presence of the superoxide-generating agents methyl viologen and 2,3-dimethoxy-1,4-naphthoquinone. Structural and functional studies showed that Xac-FNR displayed different functional features from other subclass I bacterial FNRs. Our analyses suggest that these differences may be due to the unusual carboxy-terminal region. We propose a further classification of subclass I bacterial FNRs, which is useful to determine the nature of their ferredoxin redox partners. Using sequence analysis, we identified a ferredoxin (XAC1762 as a potential substrate of Xac-FNR. The purified ferredoxin protein displayed the typical broad UV-visible spectrum of [4Fe-4S] clusters and was able to function as substrate of Xac-FNR in the cytochrome c reductase activity. Our results suggest that Xac-FNR is involved in the oxidative stress response of Xanthomonas axonopodis pv

  12. Determination of Alternaria Brown Spot (Alternaria alternata f.sp. citri) Disease on Minneola Tangelo in Çukurova Region

    OpenAIRE

    ERKILIÇ, Ali; CANIHOŞ, Yeter; BİÇİCİ, Mehmet

    1998-01-01

    In this study, the incidence and severity of Alternaria brown spot disease (Alternaria alternata f.sp. citri) in the Çukurova region were determined. Thirty-five Minneola tangelo orchards, with a total area of 2497 da, in 24 different locations were surveyed for this purpose. Leaf infections in all orchards were observed in the spring flushing period. Disease severity was found to be 0-10 % in 10 orchards, 11-20 % in 10 orchards, 21-30 % in 7 orchards, 31-40 % in 5 orchards, 42.5 % in 1 orch...

  13. Sobre o condicionamento alimentar na cochonilha-branca, 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

  14. New occupational allergen in citrus farmers: citrus red mite (Panonychus citri).

    Science.gov (United States)

    Kim, Y K; Son, J W; Kim, H Y; Park, H S; Lee, M H; Cho, S H; Min, K U; Kim, Y Y

    1999-02-01

    There have been several reports of occupational allergy to spider mites (Tetranychidae), but no published report has described citrus red mite (CRM, Panonychus citri)-induced occupational asthma confirmed by specific bronchial challenge. The purpose of this study was to evaluate clinical and immunologic characteristics of CRM-induced occupational asthma. We encountered 16 cases of CRM-induced occupational asthma among farmers cultivating citrus fruits. Asthmatic attacks corresponded closely with their work on citrus farms. The mean duration of the latent period was 12.9 (range 7 to 20) years. During their first visit to our clinic, nine patients with FEV1 lower than 70% of predictive value showed reversible airway obstruction after inhalation of bronchodilator, and seven with FEV1 greater than 70% of predictive value showed airway hyperresponsiveness to methacholine. Fifteen of the 16 also complained of recurrent nasal symptoms, which had developed at an earlier time than the asthmatic symptoms. They showed strong positive reactions to CRM extract on skin prick test (A/H ratio > or = 1.0) and had high serum specific IgE antibody against CRM which was detected by ELISA. Skin prick test with common inhalant allergens revealed that 10 had an isolated positive response to CRM with negative results to common inhalant allergens in their environment. The ELISA inhibition tests with CRM demonstrated significant inhibitions by CRM in a dose-dependent manner, while minimal inhibitions were noted by D. pteronyssinus and mugwort allergens. These findings suggest that CRM could induce IgE-mediated bronchoconstriction in exposed workers on citrus farm.

  15. Overexpression of a Gene Encoding a Catabolite Repression Element in Alternaria citri Causes Severe Symptoms of Black Rot in Citrus Fruit.

    Science.gov (United States)

    Katoh, H; Ohtani, K; Yamamoto, H; Akimitsu, K

    2007-05-01

    ABSTRACT A gene (AcCreA) encoding a catabolite repression element (CreA) with (two zinc fingers of the Cys(2)His(2) type was isolated from the postharvest fungal pathogen Alternaria citri. The AcCreA overexpression mutant AcOEC2 of A. citri showed normal growth on pectin medium and on segments of peel or the juice sac area from citrus fruit. Production of endopolygalacturonase, an essential virulence factor of this pathogen, was similar in AcOEC2 and the wild type in pectin-containing media. However, addition of glucose to the medium showed that carbon catabolite repression of endopolygalacturonase gene (Acpg1) expression, as well as endopolygalacturonase production, was lost in AcOEC2. The wild-type strain of A. citri causes rot mainly in the central axis of citrus fruit without development of rotting in the juice sac area; however, AcOEC2 caused severe black rot symptoms in both the central axis and juice sac areas. These results indicate that AcCreA-mediated catabolite repression controls the virulence or infection of this pathogen, and that the wild-type A. citri does not cause symptoms in the juice sac area due to carbon catabolite repression by sugars in the juice of the juice sac area.

  16. Differential expression of pathogenicity- and virulence-related genes of Xanthomonas axonopodis pv. citri under copper stress

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    Ana Carolina Basílio Palmieri

    2010-01-01

    Full Text Available In this study, we used real-time quantitative PCR (RT-qPCR to evaluate the expression of 32 genes of Xanthomonas axonopodis pv. citri related to pathogenicity and virulence that are also involved in copper detoxification. Nearly all of the genes were up-regulated, including copA and copB. Two genes homologous to members of the type II secretion system (xcsH and xcsC and two involved in the degradation of plant cell wall components (pglA and pel were the most expressed in response to an elevated copper concentration. The type II secretion system (xcs operon and a few homologues of proteins putatively secreted by this system showed enhanced expression when the bacteria were exposed to a high concentration of copper sulfate. The enhanced expression of the genes of secretion II system during copper stress suggests that this pathway may have an important role in the adaptative response of X. axonopodis pv. citri to toxic compounds. These findings highlight the potential role of these genes in attenuating the toxicity of certain metals and could represent an important means of bacterial resistance against chemicals used to control diseases.

  17. Functional analysis of a chitinase gene during the larval-nymph transition in Panonychus citri by RNA interference.

    Science.gov (United States)

    Xia, Wen-Kai; Shen, Xiao-Min; Ding, Tian-Bo; Niu, Jin-Zhi; Zhong, Rui; Liao, Chong-Yu; Feng, Ying-Cai; Dou, Wei; Wang, Jin-Jun

    2016-09-01

    Chitinases are hydrolytic enzymes that are required for chitin degradation and reconstruction in arthropods. In this study, we report a cDNA sequence encoding a putative chitinase (PcCht1) from the citrus red mite, Panonychus citri. The PcCht1 (564 aa) possessed a signal peptide, a conserver domain, and a chitin-binding domain. Structural and phylogenetic analyses found that PcCht1 had high sequence similarity to chitinases in Tetranychus urticae. Real-time quantitative PCR analyses showed that the transcript levels of PcCht1 peaked periodically in larval and nymph stages. Moreover, significant increase of PcCht1 transcript level in the larvae was observed upon the exposure of diflubenzuron. In contrast, exposures of the larvae to diflubenzuron resulted in the decreased chitin content. Furthermore, through a feeding-based RNA interference approach, we were able to reduce the PcCht1 transcript level by 59.7 % in the larvae, and consequently the treated larvae showed a very low molting rate compared with the control. Our results expanded the understanding of the important role of PcCht1 in the growth and development of P. citri.

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

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    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.

  19. 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.

  20. 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.

  1. The Phloem-Sap Feeding Mealybug (Ferrisia virgata) Carries ‘Candidatus Liberibacter asiaticus’ Populations That Do Not Cause Disease in Host Plants

    Science.gov (United States)

    Pitino, Marco; Hoffman, Michele T.; Zhou, Lijuan; Hall, David G.; Stocks, Ian C.; Duan, Yongping

    2014-01-01

    ‘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 (8.24×107 to 1.07×104 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. PMID:24465578

  2. Antitussive, expectorant and anti-inflammatory activities of different extracts from Exocarpium Citri grandis.

    Science.gov (United States)

    Jiang, Kun; Song, Qian; Wang, Lei; Xie, Tianzhu; Wu, Xi; Wang, Ping; Yin, Guo; Ye, Wencai; Wang, Tiejie

    2014-10-28

    Exocarpium Citri grandis (C. grandis, Huajuhong in Chinese), the epicarp of C. grandis 'Tomentosa', is used as an antitussive, expectorant and anti-inflammatory drug for hundreds of years in China. The study was aimed at evaluating the antitussive, expectorant and anti-inflammatory effects of different extracts of C. grandis, providing experimental evidence for its traditional use, and laying a foundation for its further researches. Crude drugs of C. grandis were extracted with four kinds of solvents (water, 50% ethanol, 70% ethanol and 90% ethanol) in reflux conditions, respectively. Solutions were concentrated in reduced pressure and lyophilized in vacuum to yield the aqueous extract, 50% ethanolic extract, 70% ethanolic extract, and 90% ethanolic extract of C. grandis. Antitussive evaluations were carried out with ammonia liquor induced mice cough; expectorant effects were tested with phenol red secretion experiments in mice; anti-inflammatory effects were assessed by murine model of xylene induced ear edema in mice. Only aqueous and 70% ethanolic extracts of C. grandis displayed significant antitussive, expectorant and anti-inflammatory activities. Aqueous extract of C. grandis significantly decreased cough frequency caused by ammonia liquor, increased phenol red secretion and inhibited the development of ear edema in anti-inflammatory assay at the dose of 1005 mg/kg (Pstrong effect of decreasing cough frequency, prolonging cough period, increasing phenol red secretion and decreasing the extent of ear edema at the dose of 493 mg/kg (P<0.001). The low, middle, and high dose (247, 493, and 986 mg/kg) of 70% ethanolic extract of C. grandis showed significant antitussive, expectorant and anti-inflammatory effects in good dose dependant manner. The results supported the folk use of C. grandis (decoction of C. grandis) with scientific evidence, and indicated that the 70% ethanolic extract of C. grandis might have better effects of antitussive, expectorant and anti

  3. The complete mitochondrial genome of the citrus red mite Panonychus citri (Acari: Tetranychidae): high genome rearrangement and extremely truncated tRNAs.

    Science.gov (United States)

    Yuan, Ming-Long; Wei, Dan-Dan; Wang, Bao-Jun; Dou, Wei; Wang, Jin-Jun

    2010-10-23

    The family Tetranychidae (Chelicerata: Acari) includes ~1200 species, many of which are of agronomic importance. To date, mitochondrial genomes of only two Tetranychidae species have been sequenced, and it has been found that these two mitochondrial genomes are characterized by many unusual features in genome organization and structure such as gene order and nucleotide frequency. The scarcity of available sequence data has greatly impeded evolutionary studies in Acari (mites and ticks). Information on Tetranychidae mitochondrial genomes is quite important for phylogenetic evaluation and population genetics, as well as the molecular evolution of functional genes such as acaricide-resistance genes. In this study, we sequenced the complete mitochondrial genome of Panonychus citri (Family Tetranychidae), a worldwide citrus pest, and provide a comparison to other Acari. The mitochondrial genome of P. citri is a typical circular molecule of 13,077 bp, and contains the complete set of 37 genes that are usually found in metazoans. This is the smallest mitochondrial genome within all sequenced Acari and other Chelicerata, primarily due to the significant size reduction of protein coding genes (PCGs), a large rRNA gene, and the A + T-rich region. The mitochondrial gene order for P. citri is the same as those for P. ulmi and Tetranychus urticae, but distinctly different from other Acari by a series of gene translocations and/or inversions. The majority of the P. citri mitochondrial genome has a high A + T content (85.28%), which is also reflected by AT-rich codons being used more frequently, but exhibits a positive GC-skew (0.03). The Acari mitochondrial nad1 exhibits a faster amino acid substitution rate than other genes, and the variation of nucleotide substitution patterns of PCGs is significantly correlated with the G + C content. Most tRNA genes of P. citri are extremely truncated and atypical (44-65, 54.1 ± 4.1 bp), lacking either the T- or D-arm, as found in P. ulmi

  4. The complete mitochondrial genome of the citrus red mite Panonychus citri (Acari: Tetranychidae: high genome rearrangement and extremely truncated tRNAs

    Directory of Open Access Journals (Sweden)

    Dou Wei

    2010-10-01

    Full Text Available Abstract Background The family Tetranychidae (Chelicerata: Acari includes ~1200 species, many of which are of agronomic importance. To date, mitochondrial genomes of only two Tetranychidae species have been sequenced, and it has been found that these two mitochondrial genomes are characterized by many unusual features in genome organization and structure such as gene order and nucleotide frequency. The scarcity of available sequence data has greatly impeded evolutionary studies in Acari (mites and ticks. Information on Tetranychidae mitochondrial genomes is quite important for phylogenetic evaluation and population genetics, as well as the molecular evolution of functional genes such as acaricide-resistance genes. In this study, we sequenced the complete mitochondrial genome of Panonychus citri (Family Tetranychidae, a worldwide citrus pest, and provide a comparison to other Acari. Results The mitochondrial genome of P. citri is a typical circular molecule of 13,077 bp, and contains the complete set of 37 genes that are usually found in metazoans. This is the smallest mitochondrial genome within all sequenced Acari and other Chelicerata, primarily due to the significant size reduction of protein coding genes (PCGs, a large rRNA gene, and the A + T-rich region. The mitochondrial gene order for P. citri is the same as those for P. ulmi and Tetranychus urticae, but distinctly different from other Acari by a series of gene translocations and/or inversions. The majority of the P. citri mitochondrial genome has a high A + T content (85.28%, which is also reflected by AT-rich codons being used more frequently, but exhibits a positive GC-skew (0.03. The Acari mitochondrial nad1 exhibits a faster amino acid substitution rate than other genes, and the variation of nucleotide substitution patterns of PCGs is significantly correlated with the G + C content. Most tRNA genes of P. citri are extremely truncated and atypical (44-65, 54.1 ± 4.1 bp, lacking

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

    Science.gov (United States)

    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

  6. Biocontrol ability and putative mode of action of yeasts against Geotrichum citri-aurantii in citrus fruit.

    Science.gov (United States)

    Ferraz, Luriany Pompeo; Cunha, Tatiane da; da Silva, Aline Caroline; Kupper, Katia Cristina

    2016-01-01

    Sour rot is a major postharvest disease of citrus fruit and is caused by the fungal pathogen Geotrichum citri-aurantii. A lack of chemicals certified for the control of this disease has led to the consideration of alternative methods and strategies, such as the use of yeasts as biocontrol agents. The purpose of the present study was to test the ability of yeasts isolated from leaves, flowers, fruit, and soil, and six Saccharomyces cerevisiae isolates to control citrus sour rot, to assess the mechanisms of action of the yeast isolates that were demonstrated to be effective for biocontrol, and to identify the most effective yeast isolates for the biocontrol of G. citri-aurantii. In in vivo assays, three yeast isolates (ACBL-23, ACBL-44, and ACBL-77) showed a potential for controlling sour rot in citrus fruits, both preventatively and curatively. Most of the eight yeast isolates that were assessed for a mechanism of action did not produce antifungal compounds in an amount sufficient to inhibit the growth of the pathogen. Additionally, nutrient competition among the yeast strains was not found to be a biocontrol strategy. Instead, killer activity and hydrolytic enzyme production were identified as the major mechanisms involved in the biocontrol activity of the yeasts. Isolates ACBL-23, ACBL-44, and ACBL-77, which controlled sour rot most effectively, were identified as Rhodotorula minuta, Candida azyma, and Aureobasidium pullulans, respectively. To our knowledge, this is the first report of the potential of C. azyma as a biological control agent against a postharvest pathogen and its ability to produce a killer toxin.

  7. Molecular Characterization of Vitellogenin and Its Receptor Genes from Citrus Red Mite, Panonychus citri (McGregor

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    Rui Zhong

    2015-03-01

    Full Text Available The production and uptake of yolk protein play an important role in the reproduction of all oviparous organisms. Vitellogenin (Vg is the precursor of vitellin (Vn, which is the major egg storage protein, and vitellogenin receptor (VgR is a necessary protein for the uptake of Vg into developing oocytes. In this paper, we characterize the full-length Vg and VgR, PcVg1 and PcVgR, respectively, of the citrus red mite Panonychus citri (McGregor. The PcVg1 cDNA is 5748 nucleotides (nt with a 5553-nt open reading frame (ORF coding for 1851 amino acids (aa, and the PcVgR is 6090 nt, containing an intact ORF of 5673 nt coding an expected protein of 1891 aa. The PcVg1 aa sequence shows a typical GLCG domain and several K/RXXR cleavage sites, and PcVgR comprises two ligand-binding domains, two epidermal growth factor (EGF-like regions containing YWTD motifs, a transmembrane domain, and a cytoplasmic domain. An analysis of the aa sequences and phylogenetics implied that both genes were genetically distinct from those of ticks and insects. The transcriptional profiles determined by real-time quantitative PCR in different developmental stages showed that both genes present the same expressional tendencies in eggs, larvae, nymphs, and adults. This suggested that the biosynthesis and uptake of PcVg occurs coordinately. The strong reproductive capacity of P. citri has been hypothesized as an important factor in its resistance; consequently, understanding the molecular mechanisms regulating Vg and VgR are fundamental for mite control.

  8. RNA Interference towards the Potato Psyllid, Bactericera cockerelli, Is Induced in Plants Infected with Recombinant Tobacco mosaic virus (TMV.

    Directory of Open Access Journals (Sweden)

    Hada Wuriyanghan

    Full Text Available The potato/tomato psyllid, Bactericera cockerelli (B. cockerelli, is an important plant pest and the vector of the phloem-limited bacterium Candidatus Liberibacter psyllaurous (solanacearum, which is associated with the zebra chip disease of potatoes. Previously, we reported induction of RNA interference effects in B. cockerelli via in vitro-prepared dsRNA/siRNAs after intrathoracic injection, and after feeding of artificial diets containing these effector RNAs. In order to deliver RNAi effectors via plant hosts and to rapidly identify effective target sequences in plant-feeding B. cockerelli, here we developed a plant virus vector-based in planta system for evaluating candidate sequences. We show that recombinant Tobacco mosaic virus (TMV containing B. cockerelli sequences can efficiently infect and generate small interfering RNAs in tomato (Solanum lycopersicum, tomatillo (Physalis philadelphica and tobacco (Nicotiana tabacum plants, and more importantly delivery of interfering sequences via TMV induces RNAi effects, as measured by actin and V-ATPase mRNA reductions, in B. cockerelli feeding on these plants. RNAi effects were primarily detected in the B. cockerelli guts. In contrast to our results with TMV, recombinant Potato virus X (PVX and Tobacco rattle virus (TRV did not give robust infections in all plants and did not induce detectable RNAi effects in B. cockerelli. The greatest RNA interference effects were observed when B. cockerelli nymphs were allowed to feed on leaf discs collected from inoculated or lower expanded leaves from corresponding TMV-infected plants. Tomatillo plants infected with recombinant TMV containing B. cockerelli actin or V-ATPase sequences also showed phenotypic effects resulting in decreased B. cockerelli progeny production as compared to plants infected by recombinant TMV containing GFP. These results showed that RNAi effects can be achieved in plants against the phloem feeder, B. cockerelli, and the TMV

  9. RNA Interference towards the Potato Psyllid, Bactericera cockerelli, Is Induced in Plants Infected with Recombinant Tobacco mosaic virus (TMV).

    Science.gov (United States)

    Wuriyanghan, Hada; Falk, Bryce W

    2013-01-01

    The potato/tomato psyllid, Bactericera cockerelli (B. cockerelli), is an important plant pest and the vector of the phloem-limited bacterium Candidatus Liberibacter psyllaurous (solanacearum), which is associated with the zebra chip disease of potatoes. Previously, we reported induction of RNA interference effects in B. cockerelli via in vitro-prepared dsRNA/siRNAs after intrathoracic injection, and after feeding of artificial diets containing these effector RNAs. In order to deliver RNAi effectors via plant hosts and to rapidly identify effective target sequences in plant-feeding B. cockerelli, here we developed a plant virus vector-based in planta system for evaluating candidate sequences. We show that recombinant Tobacco mosaic virus (TMV) containing B. cockerelli sequences can efficiently infect and generate small interfering RNAs in tomato (Solanum lycopersicum), tomatillo (Physalis philadelphica) and tobacco (Nicotiana tabacum) plants, and more importantly delivery of interfering sequences via TMV induces RNAi effects, as measured by actin and V-ATPase mRNA reductions, in B. cockerelli feeding on these plants. RNAi effects were primarily detected in the B. cockerelli guts. In contrast to our results with TMV, recombinant Potato virus X (PVX) and Tobacco rattle virus (TRV) did not give robust infections in all plants and did not induce detectable RNAi effects in B. cockerelli. The greatest RNA interference effects were observed when B. cockerelli nymphs were allowed to feed on leaf discs collected from inoculated or lower expanded leaves from corresponding TMV-infected plants. Tomatillo plants infected with recombinant TMV containing B. cockerelli actin or V-ATPase sequences also showed phenotypic effects resulting in decreased B. cockerelli progeny production as compared to plants infected by recombinant TMV containing GFP. These results showed that RNAi effects can be achieved in plants against the phloem feeder, B. cockerelli, and the TMV-plant system will

  10. Monitoramento da expressão gênica de Xanthomonas axonopodis pv. citri em diferentes condições ambientais

    OpenAIRE

    2003-01-01

    The bacterium Xanthomonas axonopodis pv. citri is a pathogen causing citric canker that compromises part of the orange production in the State of São Paulo, with negative consequences for the economy of the State and the Country. For the future control of citric canker, several investigators participated in the Xanthomonas Genome Project in order to determine the gene set of this bacterium and to contribute to the identification of the genes involved in its pathogenicity. In this respect, the...

  11. 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.

  12. Exposure to Diflubenzuron Results in an Up-Regulation of a Chitin Synthase 1 Gene in Citrus Red Mite, Panonychus citri (Acari: Tetranychidae

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    Wen-Kai Xia

    2014-02-01

    Full Text Available Chitin synthase synthesizes chitin, which is critical for the arthropod exoskeleton. In this study, we cloned the cDNA sequences of a chitin synthase 1 gene, PcCHS1, in the citrus red mite, Panonychus citri (McGregor, which is one of the most economically important pests of citrus worldwide. The full-length cDNA of PcCHS1 contains an open reading frame of 4605 bp of nucleotides, which encodes a protein of 1535 amino acid residues with a predicted molecular mass of 175.0 kDa. A phylogenetic analysis showed that PcCHS1 was most closely related to CHS1 from Tetranychus urticae. During P. citri development, PcCHS1 was constantly expressed in all stages but highly expressed in the egg stage (114.8-fold higher than in the adult. When larvae were exposed to diflubenzuron (DFB for 6 h, the mite had a significantly high mortality rate, and the mRNA expression levels of PcCHS1 were significantly enhanced. These results indicate a promising use of DFB to control P. citri, by possibly acting as an inhibitor in chitin synthesis as indicated by the up-regulation of PcCHS1 after exposure to DFB.

  13. Construction of EGFP-labeling system for visualizing the infection process of Xanthomonas axonopodis pv. citri in planta.

    Science.gov (United States)

    Liu, Li-Ping; Deng, Zi-Niu; Qu, Jin-Wang; Yan, Jia-Wen; Catara, Vittoria; Li, Da-Zhi; Long, Gui-You; Li, Na

    2012-09-01

    Xanthomonas axonopodis pv. citri (Xac) is the causal agent of citrus bacterial canker, an economically important disease to world citrus industry. To monitor the infection process of Xac in different citrus plants, the enhanced green florescent protein (EGFP) visualizing system was constructed to visualize the propagation and localization in planta. First, the wild-type Xac was isolated from the diseased leaves of susceptible 'Bingtang' sweet orange, and then the isolated Xac was labeled with EGFP by triparental mating. After PCR identification, the growth kinetics and pathogenicity of the transformants were analyzed in comparison with the wild-type Xac. The EGFP-labeled bacteria were inoculated by spraying on the surface and infiltration in the mesophyll of 'Bingtang' sweet orange leaves. The bacterial cell multiplication and diffusion processes were observed directly under confocal laser scanning microscope at different intervals after inoculation. The results indicated that the EGFP-labeled Xac releasing clear green fluorescence light under fluorescent microscope showed the infection process and had the same pathogenicity as the wild type to citrus. Consequently, the labeled Xac demonstrated the ability as an efficient tool to monitor the pathogen infection.

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

    Science.gov (United States)

    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.

  15. Transmission and Propagation of 'Candidatus Liberibacter asiaticus' by Grafting with Individual Citrus Leaves.

    Science.gov (United States)

    Hilf, Mark E; Lewis, Reid S

    2016-05-01

    Huanglongbing (HLB) is a chronic, progressive decline disease in citrus associated with a systemic infection by the bacterium 'Candidatus Liberibacter asiaticus'. Transmission of the bacterium in the field is by the Asian citrus psyllid, Diaphorina citri Kuwayama. Experimental propagation of 'Ca. L. asiaticus' is done primarily by grafting pieces of bud wood from an infected plant. To produce a small-scale model system for investigation of pathogen biology, we investigated grafting single leaves from infected citrus plants as sources of inoculum for propagation of the bacterium. In total, 162 plants ranging in age from 3 to 18 months were grafted. Grafting with intact asymptomatic and HLB-symptomatic leaves resulted in 61 of 78 (78%) and 35 of 41 (85%) of the plants infected with 'Ca. L. asiaticus', respectively. Inoculum consisting of the leaf petiole only or only an inoculum tissue remnant under the bark of the receptor tree resulted in 6 of 12 (50%) and 7 of 31 (23%) infected trees, respectively. Real-time polymerase chain reaction (qPCR) assays verified the infection in plants, a majority of which developed the foliar blotchy mottle symptom considered diagnostic for HLB, while some plants also displayed the stunted, chlorotic shoots for which the disease is named. The qPCR data together with the symptoms displayed demonstrated that individual leaves from infected trees can serve as effective inoculum sources for transmission and propagation of 'Ca. L. asiaticus' via grafting.

  16. A review of techniques for detecting Huanglongbing (greening) in citrus.

    Science.gov (United States)

    Arredondo Valdés, Roberto; Delgado Ortiz, Juan C; Beltrán Beache, Mariana; Anguiano Cabello, Julia; Cerna Chávez, Ernesto; Rodríguez Pagaza, Yolanda; Ochoa Fuentes, Yisa M

    2016-10-01

    Huanglongbing (HLB) is the most destructive disease of citrus worldwide. Monitoring of health and detection of diseases in trees is critical for sustainable agriculture. HLB symptoms are virtually the same wherever the disease occurs. The disease is caused by Candidatus Liberibacter spp., vectored by the psyllids Diaphorina citri Kuwayama and Trioza erytreae. Electron microscopy was the first technique used for HLB detection. Nowadays, scientists are working on the development of new techniques for a rapid HLB detection, as there is no sensor commercially accessible for real-time assessment of health conditions in trees. Currently, the most widely used mechanism for monitoring HLB is exploration, which is an expensive, labor-intensive, and time-consuming process. Molecular techniques such as polymerase chain reaction are used for the identification of HLB disease, which requires detailed sampling and processing procedures. Furthermore, investigations are ongoing in spectroscopic and imaging techniques, profiling of plant volatile organic compounds, and isothermal amplification. This study recognizes the need for developing a rapid, cost-effective, and reliable health-monitoring sensor that would facilitate advancements in HLB disease detection. This paper compares the benefits and limitations of these potential methods for HLB detection.

  17. 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

    2016-06-02

    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.

  18. The Effect of Ultraviolet-A Radiation Exposure on the Reproductive Ability, Longevity, and Development of the Dialeurodes citri (Homoptera: Aleyrodidae) F1 Generation.

    Science.gov (United States)

    Tariq, Kaleem; Noor, Mah; Saeed, Shafqat; Zhang, Hongyu

    2015-12-01

    Ultraviolet (UV) light has been used worldwide to monitor and trap insect pests. Whitefly adults show conspicuous positive phototactic behavior toward UV light stimuli; however, knowledge of the effect of UV light exposure on various life-history parameters of Dialeurodes citri remains limited. The present research aimed to investigate the effect of ultraviolet radiation (UV-A; long-wave) exposure on the reproduction and longevity of D. citri adults as well as the development of immature (eggs, larvae, and pupae) flies in the F1 generation. Paired D. citri adults were exposed to UV-A radiation for different periods (0, 1, 4, and 7 h/d) until the end of their life. The results of the experiment revealed that fecundity and oviposition rates increased when adults were irradiated for 1 and 4 h/d, but interestingly, both were significantly decreased compared with those of the controls after the longest exposure time (7 h/d). The longevity of adults of both sexes and the cumulative survival of F1 immatures were decreased with increased exposure time. Exposure to UV-A radiation prolonged the developmental time of immature stages, and a positive correlation was observed with exposure time. Exposure to UV light significantly inhibited egg hatching, larval development, pupation, and adult emergence. To the best of our knowledge, this is the first study describing the effect of UV radiation on a homopteran insect pest. This research may provide a foundation for the scientific community to use UV light in the field as an integrated pest management strategy to control this devastating agricultural pest.

  19. In silico identification of potential chaperone genes that belong to type III and type IV secretion systems in Xanthomonas axonopodis pv citri

    Directory of Open Access Journals (Sweden)

    Letícia Khater

    2005-01-01

    Full Text Available The secretion of bacterial virulence factors and flagellar components requires the assistance of specific type III and flagellar chaperones. Standard computational annotation of the genome of Xanthomonas axonopodis pv citri, a plant pathogen that causes citrus canker, initially did not identify any genes belonging to these chaperone categories since the primary sequence homology between them was very low. However, in a search for hypothetical proteins with characteristics similar to these chaperones, we have now identified 30 chromosomal and 10 plasmidial potential genes encoding chaperones belonging to types III/IV, and flagellar secretion systems in this organism. The significance of these findings is discussed.

  20. Biologia e tabela de vida de fertilidade de Planococcus citri em diferentes estruturas vegetativas de cultivares de videira Biology and fertility life table of Planococcus citri in different vegetative structures of grape cultivars

    Directory of Open Access Journals (Sweden)

    Wilson José Morandi Filho

    2008-08-01

    Full Text Available O objetivo deste trabalho foi estudar a biologia e a tabela de vida de fertilidade de Planococcus citri, em folhas de videira das cultivares Cabernet Sauvignon, Itália e Isabel, em bagas de uva 'Itália' e em raízes da 'Isabel' e dos porta-enxertos 101-14 e IAC-572. Os seguintes parâmetros foram avaliados: duração e viabilidade dos estágios de ovo e ninfa; e fecundidade e longevidade dos adultos. Em raízes do porta-enxerto IAC-572, a cochonilha não completou o ciclo biológico. Em bagas de uva 'Itália', o inseto completou a fase de ninfa, porém os adultos foram inférteis. Em folhas das diferentes cultivares, a duração média do período de ovo a adulto dos machos foi de 24,63 dias, com viabilidade de 32%, enquanto as fêmeas duraram 32 dias com viabilidade de 56%. Em raízes, a duração do ciclo biológico de fêmeas e machos foi de 32,45 e 29,50 dias, respectivamente. Em folhas, a fecundidade foi de 67,27, 66,09 e 53,33 ovos por fêmea, nas cultivares Cabernet Sauvignon, Isabel e Itália, respectivamente. Nas raízes foram obtidos 30,4 e 70,0 ovos por fêmea, no porta-enxerto 101-14 e na cultivar Isabel, respectivamente.The objective of this work was to study the biology and fertility life table of Planococcus citri in leaves of the grape cultivars Cabernet Sauvignon, Itália and Isabel, in berries of 'Itália', in roots of 'Isabel' and 101-14 and IAC-572 rootstocks. The following parameters were evaluated: duration and viability of the egg and nymph stages; and fertility and longevity of the adults. On berries of 'Itália', the insect completed the nymph phase, however the adults were infertile. On roots of the rootstock IAC-572, citrus mealybug was unable to develop. On leaves of the grape cultivars, the period of egg to adult males was 24.63 days, with viability of 32%, while females lasted 32 days with viability of 56%. On roots, the duration of the cycle of females and males was 32.45 and 29.50 days, respectively. For females

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  3. Genome-wide mutagenesis of Xanthomonas axonopodis pv. citri reveals novel genetic determinants and regulation mechanisms of biofilm formation.

    Directory of Open Access Journals (Sweden)

    Jinyun Li

    Full Text Available Xanthomonas axonopodis pv. citri (Xac causes citrus canker disease, a major threat to citrus production worldwide. Accumulating evidence suggests that the formation of biofilms on citrus leaves plays an important role in the epiphytic survival of this pathogen prior to the development of canker disease. However, the process of Xac biofilm formation is poorly understood. Here, we report a genome-scale study of Xac biofilm formation in which we identified 92 genes, including 33 novel genes involved in biofilm formation and 7 previously characterized genes, colR, fhaB, fliC, galU, gumD, wxacO, and rbfC, known to be important for Xac biofilm formation. In addition, 52 other genes with defined or putative functions in biofilm formation were identified, even though they had not previously reported been to be associated with biofilm formation. The 92 genes were isolated from 292 biofilm-defective mutants following a screen of a transposon insertion library containing 22,000 Xac strain 306 mutants. Further analyses indicated that 16 of the novel genes are involved in the production of extracellular polysaccharide (EPS and/or lipopolysaccharide (LPS, 7 genes are involved in signaling and regulatory pathways, and 5 genes have unknown roles in biofilm formation. Furthermore, two novel genes, XAC0482, encoding a haloacid dehalogenase-like phosphatase, and XAC0494 (designated as rbfS, encoding a two-component sensor protein, were confirmed to be biofilm-related genes through complementation assays. Our data demonstrate that the formation of mature biofilm requires EPS, LPS, both flagellum-dependent and flagellum-independent cell motility, secreted proteins and extracellular DNA. Additionally, multiple signaling pathways are involved in Xac biofilm formation. This work is the first report on a genome-wide scale of the genetic processes of biofilm formation in plant pathogenic bacteria. The report provides significant new information about the genetic

  4. Effects of temperature on population growth parameters of Cryptolaemus montrouzieri (Coleoptera: Coccinellidae reared on Planococcus citri (Homoptera: Pseudococcidae

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    Negar Saeedi

    2015-09-01

    Full Text Available The effect of temperature on the development, survival, fecundity and population growth parameters of Cryptolaemus montrouzieri Mulsant (Col.: Coccinellidae reared on Planococcus citri Risso (Hom.: Pseudococcidae was determined at five constant temperatures (18, 22, 26, 30 and 32±1°C in the laboratory conditions. Results showed a temperature - dependent development of the coccinellid. The mean total of immature period was estimated 76.6±0.9, 33.9±0.1, 25.8±0.1, 22.6±0.1 and 26.2±1.4 days at 18, 22, 26, 30 and 32°C, respectively. The highest and lowest values of R0, rm and λ were obtained 369.9±50.2 and 2.5±0.8 (female/female/generation, 0.07±0.01 and 0.02±0.01 (female/female/day and 1.08±0.01 and 1.02±0.01 (day-1 at 26 and 32°C, respectively. The lowest values of generation time (T and doubling time (Dt were calculated 42.2±0.9 and 8.9±0.5 days at 32 and 26°C, respectively. The lower and higher developmental threshold of total of immature period were obtained 11.5 and 25.5°C, respectively. The thermal requirement for completion of total of immature period of this predator were estimated 400 DD. These results showed that 26°C and/or adjacent temperature is most suitable for mass rearing of this predator.

  5. 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

  6. 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...

  7. Assessment of Prey Preference by the Generalist Predator, Mallada basalis (Walker), When Offered Two Species of Spider Mites, Tetranychus kanzawai Kishida and Panonychus citri (McGregor) on Papaya

    Science.gov (United States)

    We investigated potential prey preference of the generalist predator Mallada basalis (Walker) when offered two mites, Tetranychus kanzawai Kishida and Panonychus citri (McGregor), both important pests on papaya. Laboratory choice tests revealed that none of the three larval instars of M. basalis sho...

  8. Characterization and functional analysis of a novel glutathione S-transferase gene potentially associated with the abamectin resistance in Panonychus citri (McGregor).

    Science.gov (United States)

    Liao, Chong-Yu; Xia, Wen-Kai; Feng, Ying-Cai; Li, Gang; Liu, Hai; Dou, Wei; Wang, Jin-Jun

    2016-09-01

    The citrus red mite, Panonychus citri (McGregor), a major citrus pest distributed worldwide, has been found to be resistant to various insecticides and acaricides used in China. However, the molecular mechanisms associated with the abamectin resistance in this species have not yet been reported. In this study, results showed over-expression of a novel glutathione S-transferases (GSTs) gene (PcGSTm5) in abamectin-resistant P. citri. Quantitative real-time PCR analysis showed that the transcripts of PcGSTm5 were also significantly up-regulated after exposure to abamectin and the maximum mRNA expression level at nymphal stage. The recombinant protein of PcGSTm5-pET-28a produced by Escherichia coli showed a pronounced activity toward the conjugates of 1-chloro-2,4 dinitrobenzene (CDNB) and glutathione (GSH). The kinetics of CDNB and GSH and its optimal pH and thermal stability were also determined. Reverse genetic study through a new method of leaf-mediated dsRNA feeding further support a link between the expression of PcGSTm5 and abamectin resistance. However, no direct evidence was found in metabolism or inhibition assays to confirm the hypothesis that PcGSTm5 can metabolize abamectin. Finally, it is here speculated that PcGSTm5 may play a role in abamectin detoxification through other pathway such as the antioxidant protection.

  9. New Protein-Protein Interactions Identified for the Regulatory and Structural Components and Substrates of the Type III Secretion System of the Phytopathogen Xanthomonas axonopodis Pathovar citri

    Science.gov (United States)

    Alegria, Marcos C.; Docena, Cassia; Khater, Leticia; Ramos, Carlos H. I.; da Silva, Ana C. R.; Farah, Chuck S.

    2004-01-01

    We have initiated a project to identify protein-protein interactions involved in the pathogenicity of the bacterial plant pathogen Xanthomonas axonopodis pv. citri. Using a yeast two-hybrid system based on Gal4 DNA-binding and activation domains, we have focused on identifying interactions involving subunits, regulators, and substrates of the type III secretion system coded by the hrp (for hypersensitive response and pathogenicity), hrc (for hrp conserved), and hpa (for hrp associated) genes. We have identified several previously uncharacterized interactions involving (i) HrpG, a two-component system response regulator responsible for the expression of X. axonopodis pv. citri hrp operons, and XAC0095, a previously uncharacterized protein encountered only in Xanthomonas spp.; (ii) HpaA, a protein secreted by the type III secretion system, HpaB, and the C-terminal domain of HrcV; (iii) HrpB1, HrpD6, and HrpW; and (iv) HrpB2 and HrcU. Homotropic interactions were also identified for the ATPase HrcN. These newly identified protein-protein interactions increase our understanding of the functional integration of phytopathogen-specific type III secretion system components and suggest new hypotheses regarding the molecular mechanisms underlying Xanthomonas pathogenicity. PMID:15342589

  10. Research of Exocarpium Citri Grandis Fingerprint Based on Self-organization Competitive Neural Network%基于自组织竞争型神经网络的化橘红指纹图谱研究

    Institute of Scientific and Technical Information of China (English)

    陈南迪; 卢丽萍; 汪金玉; 林励

    2012-01-01

    Objective To establish the self-organization competitive neural network method for identifying the species by the fingerprints of different kinds of Exocarpium Citri Grandis(Cirtus grandis Peel). Methods The fingerprints of different species of Exocarpium Citri Grandis were established by fourier transform infrared speetroscopy and high performance liquid chromatogram (HPLC). And then we used the self-organization competitive neural network model to distinguish the different species of Exocarpium Citri Grandis. Results The self-organization competitive neural network for distinguish Exocarpium Citri Grandis of different species was established, and the rate of accuracy for infrared and HPLC fingerprint of the powder and the extract was more than 91.67 % in average. Conclusion The self-organizing competitiveneural network can be used for distinguishing different species of Exocarpium Citri Grandis by their fingerprints.%目的 建立不同品种化橘红指纹图谱的自组织竞争型人工神经网络判别方法.方法 采用傅里叶红外光谱法及HPLC法建立不同品种化橘红的指纹图谱,采用自组织竞争型人工神经网络进行模式识别.结果 自组织竞争型神经网络模型对化橘红粉末、提取物红外及HPLC指纹图谱预测平均准确率达91.67%以上.结论 自组织竞争型人工神经网络可有效用于化橘红品种的识别.

  11. Deep Characterization of the Microbiomes of Calophya spp. (Hemiptera: Calophyidae Gall-Inducing Psyllids Reveals the Absence of Plant Pathogenic Bacteria and Three Dominant Endosymbionts.

    Directory of Open Access Journals (Sweden)

    Will A Overholt

    Full Text Available Bacteria associated with sap-feeding insect herbivores include not only symbionts that may increase their hosts' fitness but also harmful plant pathogens. Calophya spp. gall-inducing psyllids (Hemiptera: Calophyidae are being investigated for their potential as biological control agents of the noxious weed, Brazilian peppertree (Schinus terebinthifolia, in Florida. Although there are no examples of plant pathogen transmission by members of the family Calophyidae, several insects in the superfamily Psylloidea are known to transmit pathogenic bacteria in the genera Candidatus Liberibacter and Candidatus Phytoplasma. To determine whether Calophya spp. harbor potentially harmful plant pathogenic bacteria, we sequenced small subunit (SSU ribosomal RNA (rRNA gene amplicons generated from individuals from four Calophya spp. populations: All microbial SSU gene sequences fell into the bacterial domain, with 98-99% belonging to the Proteobacteria. The Calophya microbiomes contained a relatively simple community, with 49-79 operational taxonomic units (OTUs; 97% detected, and only 5-8 OTUs with greater than 1% abundance. Candidatus Carsonella showed the highest relative abundance, with OTUs from this candidate genus representing between 51-65% of all recovered sequences. The next most abundant clade observed was an unclassified Enterobacteriacae group closely related to bacteria from the genera Buchnera and Blochmannia that ranged from 20-31% in relative abundance. Wolbachia populations were the third most abundant group and represented 7-27% of the diversity in microbial OTUs. No SSU rRNA gene sequences from putative pathogenic bacteria from the genera Ca. Liberibacter or Ca. Phytoplasma were detected in the microbiomes of the four Calophya populations. The probability that infected psyllids were present in our colonies, but were not sampled, was extremley low (1.39 x 10(-10. As far as we are aware, our study is the first to characterize the microbiome of

  12. The monofunctional catalase KatE of Xanthomonas axonopodis pv. citri is required for full virulence in citrus plants.

    Directory of Open Access Journals (Sweden)

    María Laura Tondo

    Full Text Available BACKGROUND: Xanthomonas axonopodis pv. citri (Xac is an obligate aerobic phytopathogen constantly exposed to hydrogen peroxide produced by normal aerobic respiration and by the plant defense response during plant-pathogen interactions. Four putative catalase genes have been identified in silico in the Xac genome, designated as katE, catB, srpA (monofunctional catalases and katG (bifunctional catalase. METHODOLOGY/PRINCIPAL FINDINGS: Xac catalase activity was analyzed using native gel electrophoresis and semi-quantitative RT-PCR. We demonstrated that the catalase activity pattern was regulated in different growth stages displaying the highest levels during the stationary phase. KatE was the most active catalase in this phase of growth. At this stage cells were more resistant to hydrogen peroxide as was determined by the analysis of CFU after the exposition to different H(2O(2 concentrations. In addition, Xac exhibited an adaptive response to hydrogen peroxide, displaying higher levels of catalase activity and H(2O(2 resistance after treatment with sub-lethal concentrations of the oxidant. In the plant-like medium XVM2 the expression of KatE was strongly induced and in this medium Xac was more resistant to H(2O(2. A XackatE mutant strain was constructed by insertional mutagenesis. We observed that catalase induction in stationary phase was lost meanwhile the adaptive response to peroxide was maintained in this mutant. Finally, the XackatE strain was assayed in planta during host plant interaction rendering a less aggressive phenotype with a minor canker formation. CONCLUSIONS: Our results confirmed that in contrast to other Xanthomonas species, Xac catalase-specific activity is induced during the stationary phase of growth in parallel with the bacterial resistance to peroxide challenge. Moreover, Xac catalases expression pattern is modified in response to any stimuli associated with the plant or the microenvironment it provides. The catalase Kat

  13. Rare diploid females coexist with rare males: a novel finding in triploid parthenogenetic populations in the psyllid Cacopsylla myrtilli (W. Wagner, 1947) (Hemiptera, Psylloidea) in northern Europe.

    Science.gov (United States)

    Nokkala, C; Kuznetsova, V G; Nokkala, S

    2015-10-01

    Using a cytological approach, diploid females were found coexisting with rare males in triploid apomictic parthenogenetic populations of the psyllid Cacopsylla myrtilli (W. Wagner, 1947) in Norway, Sweden, Finland and northwest Russia. Diploid females were easily distinguished from triploid apomictic females by the presence of 13 chiasmate bivalents instead of 39 univalent chromosomes at metaphase I. Abundance equaled that of males, but the proportion of males and diploid females was significantly greater in high altitude compared with low altitude populations. Males mated with females but showed no mating preference for diploid females. Lack of genuine bisexual reproduction owing to either asynaptic meiosis in males, or rarity of males with normal meiosis, suggests that diploids are produced in every generation by parthenogenetic females as reversals from triploidy, with their production being enhanced by environmental factor(s) associated with high altitude. This is further supported by the observation that within a population the COI haplotype found in rare males was the same as that in parthenogenetic triploid females. Thus, in northern Europe parthenogenesis in C. myrtilli is obligate, geographic parthenogenesis. Bisexual populations of C. myrtilli should be looked for in Central and Southern Europe. From the evolutionary point of view, the presence of males and diploid females with normal meiosis in parthenogenetic populations could be significant as they exhibit the potential to re-evolve either a new sexual species of parthenogenetic ancestry or a new parthenogenetic species by contagious parthenogenesis.

  14. Age at reproductive maturity and effect of age and time of day on sex attraction in the potato psyllid Bactericera cockerelli

    Institute of Scientific and Technical Information of China (English)

    Christelle Guédot; David R.Horton; Peter J.Landolt

    2012-01-01

    The potato psyllid,Bactericera (=Paratrioza) cockerelli ((S)ulc) (Hemiptera:Triozidae),is a major pest of potato.Studies were conducted to determine the age at which both males and females reach reproductive maturity and the effect of age and time of day on sex attraction.Adult B.cockerelli reach reproductive maturity within 48 h post-eclosion,with females being mature on the day of eclosion and males at 1 day post-eclosion.Oviposition generally began 2 days after mating but was delayed when females mated within 2 days post-eclosion.In laboratory olfactometer assays,the age of females used as odor sources and the age of males assayed to these odors did not affect sex attraction,with both younger (1-4-day-old) and older (8-10-day-old) males being attracted to females,regardless of female age (1-4-day-old or 8-10-day-old).Males assayed to live females at different times during the photophase (between 8:00 and 20:00 hours) were attracted to females between 11:00 and 17:00 hours,showing a temporal periodicity in sex attraction with B.cockerelli at least during the photophase.

  15. Ultrastructure of Wax-Producing Structures on the Integument of the Melaleuca Psyllid Boreioglycaspis melaleucae (Hemiptera: Psyllidae), with Honeydew Excretion Behavior in Males and Females.

    Science.gov (United States)

    Ammar, El-Desouky; Hentz, Matthew; Hall, David G; Shatters, Robert G

    2015-01-01

    The melaleuca psyllid, Boreioglycaspis melaleucae (Hemiptera: Psyllidae), was introduced to Florida as a biological control agent against Melaleuca quinquenervia, an invasive evergreen tree that has invaded large areas of Florida Everglades. Colonies of B. melaleucae nymphs are normally covered by white waxy secretions, and nymphs of various instars produce long bundles of white waxy filaments extending laterally and posteriorly from their abdomen. Scanning electron microscopy of 'naturally waxed' and 'dewaxed' nymphs (cleaned from wax) revealed two types of wax pore plates located dorsally and laterally on the integument of posterior abdominal segments starting with the 4th segment. Type-1 wax pore plates, with raised rim, peripheral groove, slits and pits, produce long ribbons and filaments of waxy secretions that are wound together forming long wax bundles, whereas type-2 wax pore plates, with slits only, produce shorter wax curls. Additionally, in both nymphs and adult females, the circumanal ring contained ornate rows of wax pores that produce wax filaments covering their honeydew excretions. Video recordings with stereomicroscopy showed that adult females produce whitish honeydew balls, powerfully propelled away from their body, probably to get these sticky excretions away from their eggs and newly hatched nymphs. Adult males, however, produce clear droplets of honeydew immediately behind them, simply by bending the posterior end of the abdomen downward. The possible role(s) of waxy secretions by nymphs and adults of B. melaleucae in reducing contamination of their colonies with honeydew, among other possibilities, are discussed.

  16. Comparison of PCR,DIA and Pathogenicity Assay for Detection of Xanthomonas axonopodis pv.citri,the Causal Agent of Citrus Bacterial Canker Disease

    Institute of Scientific and Technical Information of China (English)

    WANG Zhong-kang; SUN Xian-yun; YIN You-ping; ZHOU Chang-yong; XIA Yu-xian

    2004-01-01

    Polymerase chain reaction (PCR) approach based on newly designed primers, JYF5/JYR5, was applied for specific detection of Xanthomonas axonopodis pv.citri(Xac). The efficiency and reliability of PCR method were compared with dot immunobinding assay (DIA) and classical pathogenicity test techniques for detecting suspensions of pure cells of Xac and soaking sap of citrus tissues. Detection sensitivity of PCR was about 4.5 cells or 1.56 pg target DNA per reaction which was higher than that of DIA (ca. 450 cells per dot).These three techniques (PCR assay, DIA and Pathogenecity test) could always detect Xac from symptomatic citrus samples. Different performances were obtained from citrus materials without symptoms, and the positive detection frequency was PCR, DIA and pathogenicity test.

  17. Xanthomonas axonopodis pv. citri YaeQ structure reveals new compact protein fold built around a variation of the PD-(D/E)XK nuclease motif

    Energy Technology Data Exchange (ETDEWEB)

    Guzzo, Cristiane Rodrigues; Farah, Chuck Shaker [Universidade de Sao Paulo (USP), SP (Brazil). Inst. de Quimica. Dept. de Bioquimica; Nagem, Ronaldo Alves Pinto [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Inst. de Ciencias Biologicas. Dept. de Bioquimica e Imunologia; Barbosa, Joao Alexandre Ribeiro Goncalves [Laboratorio Nacional de Luz Sincrotron (LNLS), Campinas, SP (Brazil). Centro de Biologia Molecular Estrutural

    2008-11-15

    The YaeQ family of proteins derives from close to two hundred species of Gram-negative bacteria. Despite their widespread distribution, their molecular and physiological functions are unknown. We used X-ray crystallography and multi-wavelength anomalous dispersion to determine the high resolution structure of the YaeQ protein (XAC2398) from the phytopathogen Xanthomonas axonopodis pv. citri. The novel structure revealed that YaeQ pertains to the PD-(D/E)XK superfamily of metal-dependent endonucleases. The phylogenetic distribution of YaeQ proteins and a detailed comparison of the YaeQ structure with that of other PD-(D/E)XK family members point to specific testable hypotheses regarding YaeQ function. (author)

  18. Deciphering the regulon of a GntR family regulator via transcriptome and ChIP-exo analyses and its contribution to virulence in Xanthomonas citri.

    Science.gov (United States)

    Zhou, Xiaofeng; Yan, Qing; Wang, Nian

    2017-02-01

    Xanthomonas contains a large group of plant-associated species, many of which cause severe diseases on important crops worldwide. Six gluconate-operon repressor (GntR) family transcriptional regulators are predicted in Xanthomonas, one of which, belonging to the YtrA subfamily, plays a prominent role in bacterial virulence. However, the direct targets and comprehensive regulatory profile of YtrA remain unknown. Here, we performed microarray and high-resolution chromatin immunoprecipitation-exonuclease (ChIP-exo) experiments to identify YtrA direct targets and its DNA binding motif in X. citri ssp. citri (Xac), the causal agent of citrus canker. Integrative microarray and ChIP-exo data analysis revealed that YtrA directly regulates three operons by binding to a palindromic motif GGTG-N16 -CACC at the promoter region. A similar palindromic motif and YtrA homologues were also identified in many other bacteria, including Stenotrophomonas, Pseudoxanthomonas and Frateuria, indicating a widespread phenomenon. Deletion of ytrA in Xac abolishes bacterial virulence and induction of the hypersensitive response (HR). We found that YtrA regulates the expression of hrp/hrc genes encoding the bacterial type III secretion system (T3SS) and controls multiple biological processes, including motility and adhesion, oxidative stress, extracellular enzyme production and iron uptake. YtrA represses the expression of its direct targets in artificial medium or in planta. Importantly, over-expression of yro3, one of the YtrA directly regulated operons which contains trmL and XAC0231, induced weaker canker symptoms and down-regulation of hrp/hrc gene expression, suggesting a negative regulation in Xac virulence and T3SS. Our study has significantly advanced the mechanistic understanding of YtrA regulation and its contribution to bacterial virulence. © 2016 BSPP AND JOHN WILEY & SONS LTD.

  19. 橘全爪螨有螨叶率防治指标的初步研究%Preliminary Study on the Controlling Index of the Leaf Incidence of Panonychus citri

    Institute of Scientific and Technical Information of China (English)

    张志恒; 陈国庆; 陈亦根

    2000-01-01

    @@ 橘全爪螨(Panonychus citri Mcgregor)是为害柑橘的一种主要害虫,常因防治不及时而造成减产和品质下降.如何快速准确地实施虫情调查监测,适时指导药剂防治,是橘全爪螨综合治理的一个重要环节.

  20. All five host-range variants of Xanthomonas citri carry one pthA homolog with 17.5 repeats that determines pathogenicity on citrus, but none determine host-range variation.

    Science.gov (United States)

    Al-Saadi, Abdulwahid; Reddy, Joseph D; Duan, Yong P; Brunings, Asha M; Yuan, Qiaoping; Gabriel, Dean W

    2007-08-01

    Citrus canker disease is caused by five groups of Xanthomonas citri strains that are distinguished primarily by host range: three from Asia (A, A*, and A(w)) and two that form a phylogenetically distinct clade and originated in South America (B and C). Every X. citri strain carries multiple DNA fragments that hybridize with pthA, which is essential for the pathogenicity of wide-host-range X. citri group A strain 3213. DNA fragments that hybridized with pthA were cloned from a representative strain from all five groups. Each strain carried one and only one pthA homolog that functionally complemented a knockout mutation of pthA in 3213. Every complementing homolog was of identical size to pthA and carried 17.5 nearly identical, direct tandem repeats, including three new genes from narrow-host-range groups C (pthC), A(w) (pthAW), and A* (pthA*). Every noncomplementing paralog was of a different size; one of these was sequenced from group A* (pthA*-2) and was found to have an intact promoter and full-length reading frame but with 15.5 repeats. None of the complementing homologs nor any of the noncomplementing paralogs conferred avirulence to 3213 on grapefruit or suppressed avirulence of a group A* strain on grapefruit. A knockout mutation of pthC in a group C strain resulted in loss of pathogenicity on lime, but the strain was unaffected in ability to elicit an HR on grapefruit. This pthC- mutant was fully complemented by pthA, pthB, or pthC. Analysis of the predicted amino-acid sequences of all functional pthA homologs and nonfunctional paralogs indicated that the specific sequence of the 17th repeat may be essential for pathogenicity of X. citri on citrus.

  1. Toxicological and biochemical characterizations of AChE in phosalone-susceptible and resistant populations of the common pistachio psyllid, Agonoscena pistaciae.

    Science.gov (United States)

    Alizadeh, Ali; Talebi-Jahromi, Khalil; Hosseininaveh, Vahid; Ghadamyari, Mohammad

    2014-02-05

    The toxicological and biochemical characteristics of acetylcholinesterases (AChE) in nine populations of the common pistachio psyllid, Agonoscena pistaciae Burckhardt and Lauterer (Hemiptera: Psyllidae), were investigated in Kerman Province, Iran. Nine A. pistaciae populations were collected from pistachio orchards, Pistacia vera L. (Sapindales: Anacardiaceae), located in Rafsanjan, Anar, Bam, Kerman, Shahrbabak, Herat, Sirjan, Pariz, and Paghaleh regions of Kerman province. The previous bioassay results showed these populations were susceptible or resistant to phosalone, and the Rafsanjan population was most resistant, with a resistance ratio of 11.3. The specific activity of AChE in the Rafsanjan population was significantly higher than in the susceptible population (Bam). The affinity (K(M)) and hydrolyzing efficiency (Vmax) of AChE on acetylthiocholine iodide, butyrylthiocholine iodide, and propionylthiocholine odide as artificial substrates were clearly lower in the Bam population than that in the Rafsanjan population. These results indicated that the AChE of the Rafsanjan population had lower affinity to these substrates than that of the susceptible population. The higher Vmax value in the Rafsanjan population compared to the susceptible population suggests a possible over expression of AChE in the Rafsanjan population. The in vitro inhibitory effect of several organophosphates and carbamates on AChE of the Rafsanjan and Bam populations was determined. Based on I50, the results showed that the ratios of AChE insensitivity of the resistant to susceptible populations were 23 and 21.7-fold to monocrotophos and phosphamidon, respectively. Whereas, the insensitivity ratios for Rafsanjan population were 0.86, 0.8, 0.78, 0.46, and 0.43 for carbaryl, eserine, propoxur, m-tolyl methyl carbamate, and carbofuran, respectively, suggesting negatively correlated sensitivity to organophosphate-insensitive AChE. Therefore, AChE from the Rafsanjan population showed negatively

  2. 湖南柑橘大实蝇对不同柑橘品种的危害特点研究初报%The Damage Characteristics of Tetradacus citri Chen on Different Citrus Varieties in Hunan Province

    Institute of Scientific and Technical Information of China (English)

    林文力; 杨水芝; 潘美山; 陈海玲; 黄泽培; 龙建国; 肖伏莲

    2011-01-01

    采用定点观察和普查的方式对湖南省吉首市、麻阳县柑橘大实蝇的危害特点进行了研究.结果表明:柑橘大实蝇对不同柑橘品种的危害时间由早到晚依次为:脐橙>温洲蜜柑>冰糖橙≥大红甜橙=广柑>椪柑;其危害最严重的品种是脐橙,虫果率达63%,其次是早熟温洲蜜柑和冰糖橙,达22%,再次是椪柑,达6%;同时还发现,柑橘不同品种果皮表面存在产卵假痕迹现象.这些结果可为柑橘大实蝇的防治提供理论参考依据.%The damage characteristics of Tetradacus citri Chen in Mayang County, Jishou City, Hunan Province were investigated by fixed-point observation method and general survey method. The results indicated that the harm time of Tetradacus citri Chen on different varieties of citrus ordered as navel orange>satsuma mandarin>Citrus sinensis ^Citrus sineasis Osbeck cv. Dahong= Citrussinensis (L.) Osbeck >Citrus reticulata Blanco cv. Ponkan; the damage level of Tetradacus citri Chen on different varieties of citrus ordered as navel orange>early mature satsuma mandarin and Citrus sinensis >dtrus reticulata Blanco cv. Ponkan, and the insect infestation ratio of navel orange, early mature satsuma mandarin, Citrus sinensis and Citrus reticulata Blanco cv. Ponkan was 63%, 22%, 22% and 6%, respectively. Meanwhile, the research also showed that there are fake oviposition traces of Tetradacus citri Chen on the Peels of different varieties of citrus. These results above can provide the theory basis for the control of Tetradacus citri Chen.

  3. 橘全爪螨种群空间分布与危害%Population Spacial Distribution and Harm of Panonychus citri

    Institute of Scientific and Technical Information of China (English)

    甘宗义; 朱红艳; 潭荣荣

    2012-01-01

    The spacial distribution pattern, population dynamics and harm of Panonychus citri McG on citrus tree were studied by test of theoretical distribution and analysis of spacial pattern indexes. The results showed that the population distribution of the mite suits with Negative-binomial distribution in citrus garden, and fits with Neyman distribution-A in lower population density too. The spacial distribution pattern presents loose colonies, in which distribution of the colonies is aggregated and the individuals in the colonies show random distribution. The aggregation intensity of population distribution is changing with changes in the mite population density. The aggregation in low density population is caused by environmental factors; and in highdeasity population it is concerned with their biological activities. When the population density a certain up to a certain harm grade, the spacial volume reaching and outstripping this harm level is not over 30% of the total space in citrus garden, accordingly, the area sprayed with insecticides for controlling the harmful mite should not be over 30% of all the area in the field.%通过理论分布的适合性检验、空间格局指标分析,对橘全爪螨(Panonychus citri McG)在温州蜜柑上的空间分布特征、动态与危害进行了系统研究.结果表明,橘全爪螨田间种群分布符合负二项分布,低密度下偶尔符合Neyman A分布.空间分布的基本成分为疏松的个体群,个体群内成员间的分布为随机分布,个体群间呈聚集分布,聚集强度随种群密度的变化而变化,种群低密度下的聚集主要由环境因素引起,高密度下的聚集与该螨自身生物学特性有关.当田间种群数量达到某一危害级别时,相应级别的危害空间容积不超过总体空间的30%,相应的田间药剂防治面积也不应超过总面积的30%.

  4. Occurrence and control of citrus melanose caused by Diaporthe citri%柑橘黑点病的发病规律和防治

    Institute of Scientific and Technical Information of China (English)

    姜丽英; 徐法三; 黄振东; 黄峰; 陈国庆; 李红叶

    2012-01-01

    近年来,柑橘黑点病(Diaporthe citri)在浙江、江西等柑橘产区普遍发生,逐渐成为柑橘最主要病害之一,严重影响鲜销果实的外观品质和销售价格.2008年以来,通过对该病的症状、发生规律和防治技术开展研究发现,果实整个生长期都能感病,但以果实膨大期最感病;黑点病的严重度与树龄、施肥水平和杀菌剂的使用次数关系密切;以座果后每隔20d使用保护性杀菌剂代森锰锌防治效果较好,代森锰锌中添加0.25%~0.50%矿物油(绿颖)能有效地提高其防效.文章同时结合国内外相关研究,对黑点病症状、病原、发生规律和防治技术作了较为详尽的综述.%Melanose, caused by Diaporthe citri, has become a serious disease on citrus fruits in citrus-growing region of Zhejiang, Jiangxi provinces etc. It deteriorates exterior quality and brings down sale price of citrus fruits for fresh marketing. We have been involved in the investigation of the symptoms, prevalence and control methods of this disease since 2008. It was demonstrated that citrus fruits are susceptible to this disease in whole developmental stage, and especially susceptible during the enlarging stage. The order of melanose severity was closely associated with citrus age, times of fertilization and fungicide application. The best control efficiency could be obtained by application of protective fungicides such as mancozeb (followed the instruction of manufactory) 20 days interval after fruit-setting. Superior efficiency would be even achieved when 0. 25% to 0. 5% mineral oil ( SK spray oil) was mixed with mancozeb. Inter- and intranational information regarding the symptoms, characteristics of pathogen, epidemic factors and disease control of melanose was elaborately integrated in this paper.

  5. Aspectos biológicos de Chrysoperla externa (Hagen, 1861(Neuroptera: Chrysopidae alimentada com Planococcus citri (Risso, 1813(Hemiptera: Pseudococcidae e Toxoptera citricida (Kirkaldy, 1907 (Hemiptera: Aphididae Bipogical aspects of Chrysoperla externa (Hagen, 1861 (Neuroptera: Chrysopidae fed with Planococcus citri (Risso, 1813 (Hemiptera: Pseudococcidae and Toxoptera citricida (Kirkaldy, 1907 (Hemiptera: Aphididae (Hemiptera: Aphididae

    Directory of Open Access Journals (Sweden)

    Jean Patrick Bonani

    2009-02-01

    Full Text Available O ecossistema citrícola abriga muitas espécies de inimigos naturais, entre eles Chrysoperla externa (Hagen, 1861 que apresenta um grande potencial como agente de controle biológico de pragas. Neste trabalho, objetivou-se avapar o efeito das presas Planococcus citri (Risso, 1813 e Toxoptera citricida (Kirkaldy, 1907 sobre o desenvolvimento das fases de larva e pupa de C. externa. O ensaio foi conduzido em sala cpmatizada a uma temperatura de 25±1ºC, 70±10% UR e fotofase de 12 horas, sendo os tratamentos constituídos pelo fornecimento contínuo e alternado dessas duas presas em cada ínstar do predador, além de ovos de Anagasta kuehniella (Zeller, 1879, utilizados para fins de comparação. Foram avaliados o tempo de vida e a viabilidade das fases de larva, pupa e do período larva-adulto de C. externa. Constatou-se que T. citricida fornecido isoladamente ao longo do desenvolvimento larval ou em dois ínstares consecutivos do predador, não foi uma presa adequada, constatando-se 100% de mortalidade no período larva-adulto. Quando as larvas alimentaram-se de P. citri fornecida em dois ínstares do predador, a viabilidade no período larva-adulto foi semelhante à dieta constituída por apenas ovos de A. kuehniella, exceto quando se alimentaram de T. citricida no primeiro ínstar.The citrus-growing ecosystem houses a great deal of species of natural enemies, among them Chrysoperla externa (Hagen, 1861, which presents a great potential for use in the bipogical contrp of pests. The aim of this work was to evaluate the effects of the preys Planococcus citri (Risso, 1813 and Toxoptera citricida (Kirkaldy, 1907 on the development of larva and pupa of C. externa. The trial was carried out in an accpmatized room at 25±1ºC, 70±10% RH and 12-hour photophase, the treatments consisting of the continuous and alternate supply of those two preys in each instar of the predator, in apition to eggs of Anagasta kuehniella (Zeller, 1879 utilized for

  6. Dicty_cDB: SFB147 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 5 DQ673411_1( DQ673411 |pid:none) Diaphorina citri putative ribosoma... 48 9e-05 EU934275_1( EU934275 |pid:none) TSA: Anopheles darli...ngi AD-251 40S... 48 9e-05 EF634885_1( EF634885 |pid:none) Pristionchus sp. 10 RS51

  7. Repellent Activity of Extracts of Wild Rice Species against Panonychus citri and Aphis citricola in Associated with Esterase Isoenzyme in Insests

    Institute of Scientific and Technical Information of China (English)

    WAN Shu-qing; LIU Xiang-fa; FENG Guo-zhong; PAN Da-jian

    2006-01-01

    Six species of wild rice with different ecophenotypes including Oryza grandiglumis (E6-1, E6-3 / 6-4), O. minuta (E13-9,E13-13), O. officinalis (E15-8, E15-13), O. punctata (E16-1, E16-3, E1 6-13), O. granulata (E7-4), and O. latifolia (101392, E9-1, E9-10)were extracted with methnol and the repellent activity of the extracts against the two insects Aphis citricola and Panonychus citri were studied. The extracts of O. officinalis E15-8 showed higher repellent rate to the two insects than those of the other species. The repellent rates of the extracts of E15-8 to P. citriand A. citricola were 83.26% and 87.86% at 5×104 μg/mL in 24 h and 87.95% and 82.43% in 48 h, respectively. The extracts of O. officinalis E15-8 had the effect of inhibition to the esterase of the two insects.

  8. The filamentous phage XacF1 causes loss of virulence in Xanthomonas axonopodis pv. citri, the causative agent of citrus canker disease

    Directory of Open Access Journals (Sweden)

    Abdelmonim Ali Ahmad

    2014-07-01

    Full Text Available In this study, filamentous phage XacF1, which can infect Xanthomonas axonopodis pv. citri (Xac strains, was isolated and characterized. Electron microscopy showed that XacF1 is a member of the family Inoviridae and is about 1000 nm long and 16 nm in width. The genome of XacF1 is 7325 nucleotides in size, containing 13 predicted open reading frames (ORFs, some of which showed significant homology to Ff-like phage proteins such as ORF1 (pII, ORF2 (pV, ORF6 (pIII, and ORF8 (pVI. XacF1 showed a relatively wide host range, infecting seven out of 11 strains tested in this study. Frequently, XacF1 was found to be integrated into the genome of Xac strains. This integration occurred at the host dif site (attB and was mediated by the host XerC/D recombination system. The attP sequence was identical to that of Xanthomonas phage Cf1c. Interestingly, infection by XacF1 phage caused several physiological changes to the bacterial host cells, including lower levels of extracellular polysaccharide production, reduced motility, slower growth rate, and a dramatic reduction in virulence. In particular, the reduction in virulence suggested possible utilization of XacF1 as a biological control agent against citrus canker disease.

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

    Energy Technology Data Exchange (ETDEWEB)

    Pegos, V.R. [Universidade Estadual de Campinas (UNICAMP), SP (Brazil); Oliveira, P.S.L.; Balan, A. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil)

    2012-07-01

    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{sub 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{sub 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{sub 2}, FMN or both substrates, we characterized the residues of the pocket, the mechanism of substrate interaction and transfer of electrons from NADPH{sub 2} to FMN. This is the first report that links functional and biochemical data with structural analyses. (author)

  10. Evaluation of suitable reference genes for quantitative RT-PCR during development and abiotic stress in Panonychus citri (McGregor) (Acari: Tetranychidae).

    Science.gov (United States)

    Niu, Jin-Zhi; Dou, Wei; Ding, Tian-Bo; Yang, Li-Hong; Shen, Guang-Mao; Wang, Jin-Jun

    2012-05-01

    Quantitative real time reverse transcriptase polymerase chain reaction (RT-qPCR) is preferred for gene expression analysis in living organisms. Currently, it is a valuable tool for biological and ecological studies as it provides a relatively straightforward way to assess the relevance of transcriptional regulation under developmental and stress tolerance conditions. However, studies have shown that some commonly used reference genes varied among different experimental treatments, thus, systematic evaluation of reference genes is critical for gene expression profiling, which is often neglected in gene expression studies of arthropods. The aim of this study is to identify the suitable reference genes for RT-qPCR experiments involving various developmental stages and/or under abiotic stresses in citrus red mite Panonychus citri, a key pest in citrus orchards worldwide. GeNorm, NormFinder, and Bestkeeper software analysis indicates that elongation factor-1 alpha (ELF1A), RNA polymerase II largest subunit, alpha tublin, and glyceraldhyde-3-phosphate dehydrogenase (GAPDH) are the most stable reference genes in various developmental stages, meanwhile, ELF1A and GAPDH were the most stable reference genes under various abiotic stresses. Furthermore, this study will serve as a resource to screen reference genes for gene expression studies in any other spider mite species.

  11. Optimization of Extraction Technology for Inositol from Exocarpium Citri Grandis%正交试验法优选化橘红中肌醇提取工艺

    Institute of Scientific and Technical Information of China (English)

    陈地灵; 陈振兴; 林励; 吴泽青; 吴俊生

    2012-01-01

    [目的]优选化橘红中肌醇最佳提取工艺.[方法]以乙醇浓度、料液比和提取温度为考察因素,采用高效液相一蒸发光散射检测器(HPLC-ELSD)色谱法测定化橘红中肌醇含量,采用正交试验法优选化橘红中肌醇最优提取工艺.[结果]化橘红中肌醇最佳提取工艺为体积分数70%乙醇、料液比1∶20、提取温度100℃、每次时间45 min、提取2次.[结论]本研究优选得到的提取工艺稳定、提取率高,可用于化橘红中肌醇的提取.%Objective To optimize the extraction technology for the inositol from Exocarpium Citri Grandis. Methods With ethanol concentration, the ratio of material to solvent and extracting temperature as factors, the extraction technology for the inositol from Exocarpium Citri Crandis was optimized by orthogonal design and through the determination of inositol content by high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD). Results The optimal extraction technology was as follows; 70% ethanol, the ratio of material to solvent being 1:20, extraction at 100℃, for 45 min and extraction twice. Conclusion The extraction technology is steady, extraction percentage is higher and the technology is suitable for the extraction of inositol from Exocarpium Citri Crandis.

  12. Studies on Growth and Decline Law and Chemical Control of Panonychus citri in Lemon Orchard in Ruili%瑞丽柠檬园红蜘蛛消长规律及药剂防治研究

    Institute of Scientific and Technical Information of China (English)

    王自然; 段惠芬; 彭满秀; 郭俊; 赵俊; 潘黎; 高俊燕

    2014-01-01

    The growth and decline law and chemical control of Panonychus citri in lemon orchard are studied in Ruili city , Yunnan province .The results indicate that Panonychus citri occurs mainly from November to the next January , and its population density reaches a peak (2500~3000 heads per plant) in middle January;the occurrence quantity of this mite is more in the north and south of lemon crown;the occurrence and damage of Panonychus citri in lemon orchard are closely related with temperature , hu-midity, food, fertilization, chemical control and so on;among the tested 7 kinds of pesticides, 4%Spirodiclofen SC, 1.8%Abam-ectin EC and 73%Propargite EC have better control effects on Panonychus citri, and they are safe and harmless to the young shoots , flowers and young fruits of lemon .%在云南省瑞丽市进行了柠檬园红蜘蛛的消长规律及药剂防治研究。结果表明:柠檬红蜘蛛主要在11月至次年1月发生,其发生量在1月中旬达到高峰(2500~3000头/株);红蜘蛛在树冠北方、南方的发生量较多;柠檬园红蜘蛛的发生和为害与温度、湿度、食料、施肥、施药等因素关系密切;在7种供试药剂中,以4%螺螨酯悬浮剂、1.8%阿维菌素乳油和73%炔螨特乳油对红蜘蛛的防治效果较好,且对柠檬的嫩梢、花和幼果安全无害。

  13. 基于电子舌技术对不同年份的化橘红的识别%Discrimination of Exocarpium Citri Grandis in Different Years Based on Electronic Tongue Techniques

    Institute of Scientific and Technical Information of China (English)

    熊萧萧; 王鲁峰; 徐晓云; 潘思轶; 王可兴

    2012-01-01

    不同年份的化橘红功效和价格均存在差异,为了快速识别未知橘红样品的年份,采用电子舌对2001~2010年的10个样品和一个盲样进行了分析.对原始数据采用主成分分析和判别因子分析.结果表明:主成分分析可以将不同年份的样品进行有效的区分,判别因子分析可以更好地区分不同年份的样品,并能进行盲样识别.在模式识别中,判别因子分析法的识别结果均优于主成分分析法,前者2个判别因子的累计方差贡献量达到了99.929%.电子舌可以较好地区分不同年份的化橘红,改进后有望应用于化橘红的快速检测.%The effect and price vary in Exocarpium Citri Grandis of different years. Ten samples of Exocarpium Citri Grandis produced from 2001 to 2010 were analyzed with electronic tongue to discriminate their ages. The study shows that both Principal Component Analysis (PCA) and Discriminant Factor Analysis (DFA) can be used to discriminate samples of different years with the latter mathod being more accurate. In the pattern recognition, the result of DFA is also more accurate than PCA with the fact that its conmulative contribution of 2-dimentional principal component reached as high as 99.929%. In conclusion, electronic tongue can be utilized as an effective way to discriminate the age level of Exocarpium Citri Grandis. The results of this study suggest the possibility of application of the propsed method in the rapid detection and analysis of Exocarpium Citri Grandis in the process of production, sales, and other related fields.

  14. Efeito do meio de cultura, temperatura, fotoperíodo e fungicidas no crescimento micelial e no controle de Alternaria alternata f. sp. citri, causador da mancha marrom do tangor murcote

    OpenAIRE

    Colturato,Adimara Bentivoglio

    2006-01-01

    A mancha marrom de alternaria, causada pela Alternaria alternata f. sp. citri, afeta o tangor murcote causando lesões em folhas, ramos e frutos, acarretando em desfolha e manchas que depreciam o fruto comercialmente. Os objetivos deste trabalho foram verificar o efeito do meio de cultura no isolamento, da temperatura e do fotoperíodo no crescimento micelial do patógeno, avaliar a sensibilidade do patógeno à fungicidas in vitro e estabelecer o melhor produto e dose para o controle da doença no...

  15. Development and validation of a Xanthomonas axonopodis pv. citri DNA microarray platform (XACarray generated from the shotgun libraries previously used in the sequencing of this bacterial genome

    Directory of Open Access Journals (Sweden)

    Zaini Paulo A

    2010-05-01

    Full Text Available Abstract Background From shotgun libraries used for the genomic sequencing of the phytopathogenic bacterium Xanthomonas axonopodis pv. citri (XAC, clones that were representative of the largest possible number of coding sequences (CDSs were selected to create a DNA microarray platform on glass slides (XACarray. The creation of the XACarray allowed for the establishment of a tool that is capable of providing data for the analysis of global genome expression in this organism. Findings The inserts from the selected clones were amplified by PCR with the universal oligonucleotide primers M13R and M13F. The obtained products were purified and fixed in duplicate on glass slides specific for use in DNA microarrays. The number of spots on the microarray totaled 6,144 and included 768 positive controls and 624 negative controls per slide. Validation of the platform was performed through hybridization of total DNA probes from XAC labeled with different fluorophores, Cy3 and Cy5. In this validation assay, 86% of all PCR products fixed on the glass slides were confirmed to present a hybridization signal greater than twice the standard deviation of the deviation of the global median signal-to-noise ration. Conclusions Our validation of the XACArray platform using DNA-DNA hybridization revealed that it can be used to evaluate the expression of 2,365 individual CDSs from all major functional categories, which corresponds to 52.7% of the annotated CDSs of the XAC genome. As a proof of concept, we used this platform in a previously work to verify the absence of genomic regions that could not be detected by sequencing in related strains of Xanthomonas.

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

    Directory of Open Access Journals (Sweden)

    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.

  17. Effect of temperature on life history and population growth parameters of Planococcus citri (Homoptera, Pseudococcidae on coleus [Solenostemon scutellarioides (L. Codd.

    Directory of Open Access Journals (Sweden)

    Goldasteh Shila

    2009-01-01

    Full Text Available The development, life history, reproduction, and population growth parameters of Planococcus citri Risso on coleus [Solenostemon scutellarioides (L. Codd.] were studied at various temperatures ranging from 10 to 37ºC, 70±10% RH, and photoperiod length of 16: 8 h (L: D. Females and males successfully developed into adults at from 15 to 32ºC and 18 to 32ºC, respectively. All first instars died at 10, 12, and 37ºC. Lower temperatures (10, 12, and 15ºC caused higher egg mortality than did higher temperatures (32, 35, and 37ºC. At all temperatures (except 15ºC, the highest percentage of nymphal mortality was observed in the first instar. The sex ratio was female-biased between 15 and 30ºC, but there was a slightly higher number of males at 32ºC. The highest adult longevities of females and males were obtained at 18 and 25°C, respectively. The pre-oviposition, oviposition, and post-oviposition periods were significantly different at various temperatures. The highest fecundity was observed at 23ºC. The shortest and longest oviposition periods occurred at 32 and 18ºC, respectively. Maximum values of the intrinsic rate of natural increase (rm, net reproduction rate (R0, and finite rate of increase (λ and the shortest mean generation time (T and doubling time (DT were obtained at 25ºC. Our findings showed citrus mealybug performances to be highly affected by temperature.

  18. 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.

  19. Rapid screening and identification of compounds with DNA-binding activity from Folium Citri Reticulatae using on-line HPLC-DAD-MS(n) coupled with a post column fluorescence detection system.

    Science.gov (United States)

    Fu, Qingrong; Zhang, Cangman; Lin, Zongtao; Sun, Hongyang; Liang, Yi; Jiang, Haixiu; Song, Zhiling; Wang, Hong; Chen, Shizhong

    2016-02-01

    To study the interactions between natural compounds and deoxyribonucleic acid (DNA), a method has been established combining a high-performance liquid chromatography-diode array detector-multi-stage mass spectrometer with a fluorescence detector (HPLC-DAD-MS(n)-FLD). The FLD was used to monitor fluorescence intensity of the ethidium bromide-DNA (EB-DNA) complex when a compound separated by HPLC was introduced. This novel method was used to simultaneously obtain the HPLC fingerprint, UV spectra, MS(n) fragments and DNA-binding activity profile of various components in Folium Citri Reticulatae. As a result, 35 compounds were identified, of which 25 were found in the extract of Folium Citri Reticulatae for the first time, and 33 compounds showed DNA-binding activities, with the most active being feruloylhexaric and p-coumaroylhexaric acids. In addition, the precision, stability and reproducibility of this method were validated by two positive controls, quercetin and hesperidin. This new on-line method is accurate, precise and reliable for further high-throughput screening of DNA-binding compounds from food samples and other complex matrices.

  20. Study on Infrared Fingerprint of Exocarpium Citri Grandis with Different Species%化橘红药材红外指纹图谱的研究

    Institute of Scientific and Technical Information of China (English)

    卢丽萍; 林励; 黄爱华; 欧立娟; 汪金玉

    2011-01-01

    Objective To provide theoretical basis and practical methods for the rapid identification of Exocarpium Citri Grandis (ECG) with different species. Methods The fingerprints were established by fourier transform ifrared spectroscopy(FTIR), and then were analyzed by Spectrum v5.0.1 software. Results With normalization of the selected points between 1,820 cm-1 and 950 cm-1 on the FTIR fingerprints of ECG powder, significant differences were shown, and the second derivative spectra of the two different species between 1,800 cm-1 and 700 cm-1 had significant differences, which can be taken as one of the major identical characteristics to distinguish the two species. Conclusion The FTIR fingerprints is effective and feasible for the identification of the two species of ECG.%目的 建立化橘红药材的红外指纹图谱,通过考察不同品种化橘红药材的红外指纹图谱差异,为化橘红药材的快速鉴别提供理论依据和实用方法.方法 采用傅里叶红外光谱法(FTIR)建立不同品种化橘红药材红外指纹图谱,应用Spectrum v5.0.1红外软件对实验数据进行处理.结果 FTIR图谱在1820~950 cm-1范围内选点归一化处理后毛橘红和光橘红差异显著,其在1800~700 cm-1范围内二阶导数谱差异亦显著,可作为两者主要鉴别特征,经处理后的FTIR指纹图谱能有效鉴别两类化橘红药材.结论 红外指纹图谱法鉴别化橘红快速、简便、可行,可有效区分毛橘红和光橘红.

  1. 枸杞木虱啮小蜂寄生行为及生物学特性的研究%Behavior and bionomics of the ectoparasitoid Tetrastichus sp. Of the wolf berry psyllid(Paratrioza sinica)

    Institute of Scientific and Technical Information of China (English)

    段立清; 冯淑军; 李海平; 邹晓林

    2002-01-01

    枸杞木虱啮小蜂Tetrastichus sp.是枸杞木虱Paratrioza sinica Yang & Li若虫的外寄生蜂,自然寄生率可达86.5%.在温度为20~25℃的室内条件下,完成1个世代需14~20 d.成虫寿命与取食有关,取食15%的蜂蜜水,雌性成虫寿命为12~17 d,雄性成虫为4~6 d.成蜂最喜欢在3龄和4龄若虫上产卵.自然种群消长与寄主虫口密度相关.每年7月到8月下旬该蜂寄生率高, 9月份下降,10月份几乎不见.Behavior and bionomics of the ectoparasitoid Tetrastichus sp. of the wolf berry psyllid(Paratrioza sinica).

  2. 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

    Directory of Open Access Journals (Sweden)

    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

  3. 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

    Directory of Open Access Journals (Sweden)

    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

  4. Investigation on the resistance of different varieties of citrus to Xanthomonas citri%不同柑橘品种对柑橘溃疡病的抗性调查

    Institute of Scientific and Technical Information of China (English)

    宋盛英; 杨光兰; 潘盛勇; 赵春穆; 石安鸠

    2002-01-01

    通过柑橘Citrus sp.4类10个品种对柑橘溃疡病Xanthomonas citri(Hasse)Dowson抗病性的调查发现,不同的柑橘品种对柑橘溃疡病的抗病程度有明显差异,甜橙类、脐橙类普遍容易感病,属高度感病品种;宽皮橘类的抗病性最强;而柚类感病轻重不一,以台湾白柚抗病性最强,而沙田柚抗病性较弱.

  5. 柑橘全爪螨微卫星位点鉴定与信息分析%Analysis of Microsatellite Loci fromPanonychus citri Based on Enriched Microsatellite Library and Transcriptome Dataset

    Institute of Scientific and Technical Information of China (English)

    魏丹丹; 刘燕; 杜洋; 李刚; 李婷; 袁明龙; 王进军

    2016-01-01

    重复类型,占总数的53.86%;其次为两碱基核心重复类型,占总数的43.36%;而四、五、六碱基重复类型以及复合型的SSR数量均较少,且数量差异不大,共占EST-SSR总数的2.78%。柑橘全爪螨EST-SSR核心重复次数主要集中在5—10次。【结论】采用磁珠富集法,并将酶切、接头连接一步化,可提高个体微小的螨类及微小昆虫SSR的富集效率。柑橘全爪螨gSSR核心重复次数要远多于从转录组数据库获得的EST-SSR。总体而言,gSSR和EST-SSR中的三碱基重复SSR具有更好的优化率。此外,柑橘全爪螨gSSR具有微卫星家族现象。%Objective] The objective of this study is to construct the microsatellite-enriched libraries, and identify genomic- microsatellite (gSSR) from the genome of the citrus red mite,Panonychus citri. Meanwhile, a large number of gene-microsatellite (EST-SSR) markers were also indentified from the transcriptome database ofP. citri. Based on these SSR sequences, the selected SSR primer pairs were validated.[Method]On the basis of isolating high quality genomic DNA ofP. citri, enriched microsatellite libraries were constructed by streptavidin-coated magnetic beads utilizing methodologies that exploit the strong affinity between biotin and the protein streptavidin. A fast and easy protocol was proposed through a combination of two different published methods. Briefly, genomic DNA was digested by the restriction enzyme and then ligated to designed adaptors. Microsatellite-containing DNA fragments were captured by streptavidin-coated magnetic beads. The beads affinity capture of microsatellite repeats using biotinylated oligonucleotide probes. Subsequently, PCR was used to amplify the captured molecules for transferring single strand DNA to double strand DNA. The PCR products were then ligated to pGEM-T Easy vector and transformed into Trans5α competent cells. Then, clones from these libraries were screened for microsatellite content. Meanwhile

  6. The GC-MS analysis of volatile oil from Atractylodes Macrocephala Koidz,Pericarpium Citri Reticulate and their mixed samples%药对白术-陈皮挥发油成分GC-MS分析

    Institute of Scientific and Technical Information of China (English)

    杨小红; 罗琼; 胡剑卓; 陈文; 谭敏

    2012-01-01

    Objective To analysis the volatile components in herbal pair Atractylodes Macrocephala -Pericarpium Citri Reticulatae (HP AM-PCR), single herb Atractylodes Macrocephala(AM) and Pericarpium Citri Reticulatae (PCR) quantitatively. Methods Volatile oil from above materials were obtained with water reflux condensation method;oil composition was detected by gas - mass spectrometry (GC-MS) and the data was analyzed with heuristic evolving latent projection (HELP); Comparing the difference between chromatographys to identify the main chemical constituents. Results There was no new compound found in herbal pair. The compounds were from single herbs. Conclusion Chemical changes in herbal pair AM -PCR were physical compatibility.%目的:定性、定量分析药对白术-陈皮、单味药白术和陈皮的主要挥发油成分,探讨药对和单味药挥发油成分变化.方法:共水回流冷凝法分别提取单味白术、陈皮及药对挥发油、气相-质谱(GC-MS)检测,直观推导式演进特征投影(HEIP)法解析所得GC-MS数据,比较图谱差异,鉴别主要化学成分.结果:药对白术-陈皮主要组分中未发现新的结构类型化合物,挥发油化合物来源于单味药材,且主要为白术挥发油.结论:药对白术-陈皮挥发油化学成分变化属于物理配伍变化.

  7. The controlling role of Panonychus citri by "Lümanyuan" in the filed and the measurement of its acute toxicity to Carassius auratus%“绿满园”对柑桔红蜘蛛的田间防治效果及对金鱼的毒性测定

    Institute of Scientific and Technical Information of China (English)

    乔虎平; 王金亮; 江庆红; 云月利; 王维; 彭宇

    2013-01-01

    测定“绿满园”水剂对柑桔红蜘蛛Panonychus citri的田间防治效果及对非靶标生物金鱼Carassius auratus 的急性毒性.结果表明,64%“绿满园”水剂800倍和1 000倍稀释液在施药后7d和15d对柑桔红蜘蛛的防治效果均极显著地高于对照杀螨剂:15%扫螨净和57%炔螨特,其中以800倍稀释液防治效果最好,15d的防效达97.21%,800倍和1 000倍稀释液的防治效果差异不明显.金鱼的死亡率随“绿满园”浓度的升高和染毒时间的延长逐渐增加.“绿满园”水剂对金鱼的急性毒性小于阿维菌素.%The controlling role of Panonychus citri by "Lumanyuan" in the filed and the measurement of its acute toxicity to Carassius auratus were carried out. The results showed that, 15 days after spraying, the controlling roles of 800 folds and 1 000 folds dilution of 64% "Lumanyuan" to P. citri was obviously higher than 15% pyridaben and 57% propargite. 800 fold dilution of "Lumanyuan" had the best controlling role to P. citri and the controlling effect was 97. 21%. There was no significant difference on controlling effects between 800 folds and 1 000 folds dilution of "Lumanyuan" to P. citri. The mortalities of C. auratus increased with the rise of the concentration and treatment time of "Lumanyuan" and "Lumanyuan" has lower acute toxicity to C. auratus than avermectin.

  8. Regular LAMP and fast LAMP for the detection of Xanthomonas axonopodis pv.citri%柑橘溃疡病菌的普通LAMP及快速LAMP检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    张仑; 殷幼平; 吴瑜佳; 王中康

    2013-01-01

    This study is aimed to establish the regular LAMP and fast LAMP for the detection of Xanthomonas axonopodis pv.citri and make it useful for the grassroots quarantine departments.A specific region of the genome of X.axonopodis pv.citri was used for the design of regular LAMP and fast LAMP primers; the regular LAMP and fast LAMP detection system were established by optimization of reaction conditions; the specificity of the system was verified using genomic DNA of several reference strains and healthy citrus leaves by using a series of gradient dilution of DNA solution of X.axonopodis pv.citri and bacterial suspension.Bacterial suspension and DNA sensitivity of regular LAMP system reached 2.25 × 104 cfu and 2.03× 10-1 ng,respectively,while those of fast LAMP system reached 2.25 cfu and 2.03× 10-5 ng,respectively.In the specificity test,both regular LAMP and fast LAMP system showed high and identical specificity.The total time for the fast LAMP reaction was within 30 minutes,only half of that for the regular LAMP,greatly improving the efficiency of the analysis.The fast LAMP system was ten thousand times as sensitive as the regular LAMP system.Regular LAMP and fast LAMP for the detection of X.axonopodis pv.citri was successfully established.Regular LAMP and fast LAMP system provide a new,fast and easy way for the detection of X.axonopodis pv.citri.%建立柑橘溃疡病菌的普通LAMP和快速LAMP检测方法,使其能应用于基层检验检疫部门对病害的快速检测.利用柑橘溃疡病菌基因组特有的保守区域设计LAMP引物,通过优化反应条件,建立柑橘溃疡病菌的普通LAMP检测体系;在普通LAMP引物的基础上设计一对环引物,建立柑橘溃疡病菌的快速LAMP检测体系,并以多种参比菌DNA以及健康柑橘叶片基因组DNA为模板对普通LAMP和快速LAMP检测体系的特异性进行了验证,利用柑橘溃疡病菌菌液和DNA溶液梯度稀释液对普通LAMP和快速LAMP检测体系的灵敏度进

  9. 阿维菌素与印楝油对柑桔红蜘蛛的增效作用%Synergistic Effect of Abamectin and Neem Oil Against Panonychus citri Me Gregor

    Institute of Scientific and Technical Information of China (English)

    熊忠华; 李保同; 熊件妹; 张纪利

    2012-01-01

    Toxicity of abamectin and neem oil against Panonychus citri and the synergistic effect of their mixture were evaluated in the lab. It was showed that, for the contact toxicity of abamectin and neem oil to adult famales of Panonychus citri, the LCX value was 0.01 jig/mL and 6 579.87 u.g/mL, respectively, after 24 h treatment. The optimal mixture ratio of abamectin with neem oil was determined by co-toxicity factor and eo-toxicity coefficient method. The results indicated that, among the 6 combinations of theses two active ingredients, the co-toxicity factor of combination at the ratios of 1 : 100 and 1 : 125 for abamectin vs neem oil was 24.50 and 31.08, respectively. Furthermore, combination at the ratio of 1 : 125 showed the highest synergism with a co-toxicity coefficient of 160.76.%室内测定阿维菌素与印楝油对柑桔红蜘蛛的毒力及其复配组合的联合作用.毒力测定结果表明,阿维菌素与印楝油对柑桔红蜘蛛雌成螨处理后24 h的LC50分别为0.01 μg/mL和6579.87 μg/mL.以共毒因子法评价阿维菌素与印楝油6种复配组合的联合作用,结果显示,二者以1∶100和1∶125两种比例复配对柑桔红蜘蛛雌成螨的共毒因子分别为24.50和31.08,具增效作用;进一步以共毒系数法确定阿维菌素与印楝油以1∶125的比例复配增效作用最为显著,共毒系数为160.76.

  10. Biologia do psilídeo-de-concha Glycaspis brimblecombei Moore (Hemiptera, Psyllidae em Eucalyptus spp. Red gum lerp psyllid Glycaspis brimblecombei Moore (Hemiptera, Psylidae biology in Eucalyptus spp.

    Directory of Open Access Journals (Sweden)

    Daniela Cristina Firmino-Winckler

    2009-01-01

    Full Text Available Biologia do psilídeo-de-concha Glycaspis brimblecombei Moore (Hemiptera, Psyllidae em Eucalyptus spp.. Glycaspis brimblecombei Moore também conhecido por psilídeo-de-concha, se caracteriza por ser uma espécie específica ao gênero Eucalyptus L'Her. Este trabalho teve por objetivo determinar o ciclo biológico de G. brimblecombei em Eucalyptus spp. O trabalho foi conduzido em câmara climatizada (BOD, sob a temperatura de 26 °C e fotofase de 12 horas. As espécies de Eucalyptus utilizadas para o experimento foram: Eucalyptus camaldulensis, E. tereticornis, E. urophylla, E. grandis, Corymbia citriodora e um híbrido de E. grandis x E. urophylla ('urograndis'. Inicialmente foram utilizados 100 repetições (ninfas tratamento (espécies de Eucalyptus. As avaliações foram diárias. Os parâmetros biológicos avaliados foram a duração e viabilidade do estágio ninfal, longevidade dos adultos, número de posturas/fêmea, duração do período embrionário, número e viabilidade ovos, longevidade dos adultos e duração do ciclo total. Os resultados obtidos demonstraram que salvo C. citriodora que apresentou ser letal ao desenvolvimento ninfal de G. brimblecombei as demais espécies de Eucalyptus testadas oferecem condições ao desenvolvimento biológico deste psilídeo, sendo que neste trabalho E. camaldulensis mostrou-se a mais adequada.Glycaspis brimblecombei Moore, also known as red gum lerp psyllid, is characterized to be specific to the genus Eucalyptus. This work aimed to evaluate G. brimblecombei biological cycle in Eucalyptus spp. The work was accomplished in acclimatized chamber (BOD, with temperature of 26 °C and photophase of 12 hours. Eucalyptus species used in this study were: Eucalyptus camaldulensis, E. tereticornis, E. urophylla, E. grandis, Corymbia citriodora and E grandis x E. urophylla hybrid ('urograndis'. Initially 100 replications (nymphs per treatment (Eucalyptus species were prepared. The evaluations were daily and

  11. 防治橘臀纹粉蚧药剂的室内筛选%Insecticide Screening for the control of Planococcus citri(Risso)(Homoptera: Pseudococcidae) in the laboratory

    Institute of Scientific and Technical Information of China (English)

    郭莹; 刘长明; 吴梅香; 杨广

    2012-01-01

    为有效地控制橘臀纹粉蚧Planococcus citri(Risso)发生,测定了6种农药在田间使用浓度下对该虫的致死效果.结果表明,在室内条件下,吡虫啉、烯啶虫胺、啶虫脒、阿维菌素和高效氯氰菊酯对1龄若虫的防治效果较好,用药后72 h校正死亡率均在80.0%以上,而杀扑磷仅74.8%.在防治应用中,建议推荐吡虫啉,烯啶虫胺,啶虫脒3种药剂,同时应针对不同虫态适当调整用药浓度和施药时间,以达到理想的防治效果.

  12. Synergistic Effect of Three Substances on Paris polyphylla Smith against Panoychus citri McGregor%3种物质对七叶一枝花防治柑桔全爪螨的增效作用

    Institute of Scientific and Technical Information of China (English)

    周顺玉; 蒋春先; 杨群芳; 李庆

    2007-01-01

    研究了邻苯二酚、氮酮和硫酸钙3种化学物质对七叶一枝花(Paris polyphylla Smith)提取物防治柑桔全爪螨(Panoychus citri McGregor)的增效作用.结果表明:七叶一枝花乙醇、氯仿、乙酸乙酯提取物单用对柑桔全爪螨的致死中浓度(LC50)分别为1 588.537、1480.071、1 402.954 μg/ml;邻苯二酚对七叶一枝花乙醇、氯仿、乙酸乙酯提取物均有显著增效作用,LC50分别为284.506、582.295、897.323 μg/ml,分别增效5.6、2.5和1.6倍;氮酮仅对七叶一枝花乙醇提取物有增效作用,增效2.1倍;硫酸钙对七叶一枝花氯仿、乙酸乙酯提取物均有增效作用,分别增效1.4和2.1倍.

  13. Microwave-based Method for Genomic DNA Extraction and Rapid Detection Method from Xanthomonas axonopodis pv.citri%柑桔溃疡病菌基因组DNA的微波法抽提及快速检测

    Institute of Scientific and Technical Information of China (English)

    姚廷山; 周常勇; 胡军华; 冉春; 李鸿筠; 刘浩强; 肖田

    2012-01-01

    利用微波热震惊提取固体表面柑桔溃疡病基因组DNA并加以优化,所得到的基因组DNA可作为PCR反应的模板进行16SrDNA基因有效扩增.与柑桔溃疡病基园组DNA其他抽提方法相比较,微波法更适用于该病的快速检测,具有快速、筒便、费用低廉等特点,且对设备的要求不高,用特异性引物XCF/XCR可实现柑桔溃疡病菌的快速鉴定.%The optimized microwave extraction was conducted to attract the genome of citrus canker disease. The genome extraction method was fast,easy to handle with low cost The extracted DNA was suitable for PCR. The microwave-based method was suitable for fast identification compare to other methods. The method was effective, easy and fast, so it was recommended that the extraction method was applied for the detection of Xanthomonas oxonopodit pv. citri using primers XCF/XCR.

  14. Investigation on Chelidonium citri attacking Murraya paniculata and its control%橘光绿天牛危害九里香绿化球调查及防治

    Institute of Scientific and Technical Information of China (English)

    刘有莲; 黄寿昌

    2003-01-01

    九里香绿化球受橘光绿天牛Chelidonium citri Grressitt为害3~4 a后,受害株率90.7%,枝枯株率74.1%,虫口密度2.11,危害度31.5%.危害造成枝枯形成空洞平均宽度为61 cm,占树冠平均周长的14.4%;枝枯形成空洞平均深度为68.4 cm,占平均树高50.8%.植株受害与方向的关系为:植株东向受害最多,占受害总株数30%;中部枝枯形成空洞平均宽度占树冠平均周长比率最大,为22.9%;西南方向枝枯形成空洞平均深度最深,占平均高度的90.9%.植株周围阴湿,虫口密度大.

  15. 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.

    Science.gov (United States)

    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.

  16. Harm of Chelidonium citri Grressitt to Murraya paniculata (L.) Jack Balls and Economic Damage Appraisal%橘光绿天牛对九里香绿化球的危害及其经济损失评估

    Institute of Scientific and Technical Information of China (English)

    刘有莲; 黄寿昌

    2003-01-01

    分别于2002年6月27日和8月29日调查广西生态工程职业技术学院内种植的九里香[Murrayapaniculata(L.)Jack]受橘光绿天牛(Chelidomum citri Grressitt)危害的状况并进行危害价值损失评估.结果表明,九里香绿化球受害株率90.7%,枝枯株率74.1%,有虫株率75.9%,虫口密度2.11,危害度31.5%.危害造成枝枯宽度平均为61 cm,占平均树冠周长的14.4%;枝枯平均深度为68.4cm,占平均树高50.8%.植株东向受害最多,占受害总株数30%;中部枝枯宽度占树冠周长比例最大,为22.9%;西南方向枝枯深度最深,占高度的90.9%.九里香绿化球价值损失占总价值的38.7%.学院的九里香绿化球受橘光绿天牛危害严重,需要及时有效的预防与防治.

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

    Science.gov (United States)

    Killiny, Nabil; Nehela, Yasser; Hijaz, Faraj; Vincent, Christopher I

    2017-06-08

    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.

  18. Extracts of Immature Orange (Aurantii fructus immaturus) and Citrus Unshiu Peel (Citri unshiu pericarpium) Induce P-Glycoprotein and Cytochrome P450 3A4 Expression via Upregulation of Pregnane X Receptor.

    Science.gov (United States)

    Okada, Naoto; Murakami, Aki; Urushizaki, Shiori; Matsuda, Misa; Kawazoe, Kazuyoshi; Ishizawa, Keisuke

    2017-01-01

    P-glycoprotein (P-gp) and cytochrome P450 3A4 (CYP3A4) are expressed in the intestine and are associated with drug absorption and metabolism. Pregnane X receptor (PXR) is the key molecule that regulates the expression of P-gp and CYP3A4. Given that PXR activity is regulated by a variety of compounds, it is possible that unknown PXR activators exist among known medicines. Kampo is a Japanese traditional medicine composed of various natural compounds. In particular, immature orange [Aurantii fructus immaturus (IO)] and citrus unshiu peel [Citri unshiu pericarpium (CP)] are common ingredients of kampo. A previous study reported that kampo containing IO or CP decreased the blood concentration of concomitant drugs via upregulation of CYP3A4 although the mechanism was unclear. Some flavonoids are indicated to alter P-gp and CYP3A4 activity via changes in PXR activity. Because IO and CP include various flavonoids, we speculated that the activity of P-gp and CYP3A4 in the intestine may be altered via changes in PXR activity when IO or CP is administered. We tested this hypothesis by using LS180 intestinal epithelial cells. The ethanol extract of IO contained narirutin and naringin, and that of CP contained narirutin and hesperidin. Ethanol extracts of IO and CP induced P-gp, CYP3A4, and PXR expression. The increase of P-gp and CYP3A4 expression by the IO and CP ethanol extracts was inhibited by ketoconazole, an inhibitor of PXR activation. The ethanol extract of IO and CP decreased the intracellular concentration of digoxin, a P-gp substrate, and this decrease was inhibited by cyclosporine A, a P-gp inhibitor. In contrast, CP, but not IO, stimulated the metabolism of testosterone, a CYP3A4 substrate, and this was inhibited by a CYP3A4 inhibitor. These findings indicate that the ethanol extract of IO and CP increased P-gp and CYP3A4 expression via induction of PXR protein. Moreover, this induction decreased the intracellular substrate concentration.

  19. Extracts of Immature Orange (Aurantii fructus immaturus) and Citrus Unshiu Peel (Citri unshiu pericarpium) Induce P-Glycoprotein and Cytochrome P450 3A4 Expression via Upregulation of Pregnane X Receptor

    Science.gov (United States)

    Okada, Naoto; Murakami, Aki; Urushizaki, Shiori; Matsuda, Misa; Kawazoe, Kazuyoshi; Ishizawa, Keisuke

    2017-01-01

    P-glycoprotein (P-gp) and cytochrome P450 3A4 (CYP3A4) are expressed in the intestine and are associated with drug absorption and metabolism. Pregnane X receptor (PXR) is the key molecule that regulates the expression of P-gp and CYP3A4. Given that PXR activity is regulated by a variety of compounds, it is possible that unknown PXR activators exist among known medicines. Kampo is a Japanese traditional medicine composed of various natural compounds. In particular, immature orange [Aurantii fructus immaturus (IO)] and citrus unshiu peel [Citri unshiu pericarpium (CP)] are common ingredients of kampo. A previous study reported that kampo containing IO or CP decreased the blood concentration of concomitant drugs via upregulation of CYP3A4 although the mechanism was unclear. Some flavonoids are indicated to alter P-gp and CYP3A4 activity via changes in PXR activity. Because IO and CP include various flavonoids, we speculated that the activity of P-gp and CYP3A4 in the intestine may be altered via changes in PXR activity when IO or CP is administered. We tested this hypothesis by using LS180 intestinal epithelial cells. The ethanol extract of IO contained narirutin and naringin, and that of CP contained narirutin and hesperidin. Ethanol extracts of IO and CP induced P-gp, CYP3A4, and PXR expression. The increase of P-gp and CYP3A4 expression by the IO and CP ethanol extracts was inhibited by ketoconazole, an inhibitor of PXR activation. The ethanol extract of IO and CP decreased the intracellular concentration of digoxin, a P-gp substrate, and this decrease was inhibited by cyclosporine A, a P-gp inhibitor. In contrast, CP, but not IO, stimulated the metabolism of testosterone, a CYP3A4 substrate, and this was inhibited by a CYP3A4 inhibitor. These findings indicate that the ethanol extract of IO and CP increased P-gp and CYP3A4 expression via induction of PXR protein. Moreover, this induction decreased the intracellular substrate concentration. PMID:28270768

  20. Application of immunoaffinity purification technology as the pretreatment technology for traditional Chinese medicine: Its application to analysis of hesperidin and narirutin in traditional Chinese medicine preparations containing Citri reticulatae Pericarpium.

    Science.gov (United States)

    Liu, Li-Na; Wang, Ying; Jin, Hong-Yu; Ma, Shuang-Cheng; Liu, Jia-Peng

    2016-05-15

    In the present study, the feasibility of immunoaffinity chromatography (IAC) as a purification technology for the analysis of bioactive components in Traditional Chinese Medicine (TCM) was evaluated. IAC was used to analyze hesperidin (HP) and narirutin (NR) in TCM preparations containing Citri reticulatae Pericarpium (CRP, Chenpi in Chinese). An IAC column for the specific extraction and enrichment of HP and NR from TCM preparations containing CRP was developed and characterized. After HP reacted with carbonyl diimidazole and coupled to protein, it was used to immune mice for the generation of antibody. Through cell fusion, cloning and screening, monoclonal antibody was obtained. The IAC column was constructed by covalently coupling specific monoclonal antibody against HP and NR to CNBr-activated Sepharose 4B and packed into a common solid phase extraction cartridge. The extraction conditions including loading, washing and eluting, as well as flow rate for the extraction of HP and NR were optimized. Under optimal conditions, the maximum capacity, extraction recovery rate and stability of IAC column was also characterized. Results revealed that the maximum capacity of IAC column for HP and NR was approximately 16μg and the relative binding capacity per 1mL of the column volume was 27μg. The extraction recovery rate of IAC column for HP and NR at three spiked levels was in the range of 94.05-109.15%. After the repeated application for 5 times, no significant loss of specific recognition was observed. Using high performance liquid chromatography (HPLC) as an effective analytic tool, HP and NR could be successfully separated via IAC column without the inference from impurities, suggesting that the extraction of HP and NR using the prepared IAC column is feasible. The application of IAC can solve the problem of quantitative analysis due to severe interference or low content. Furthermore, pretreatment methods in different matrixes can be unified. The IAC purification

  1. Study on the Dynamic Change Law of Essential Oil Components in Citri Reticulatae Pericarpium from the Same Plant%源于同一植株陈皮挥发油成分动态变化规律研究

    Institute of Scientific and Technical Information of China (English)

    王坚; 韦正; 陈鸿平; 银玲; 刘荣; 刘友平

    2013-01-01

    目的:研究源于同一植株不同生长期陈皮挥发油成分的变化规律.方法采用水蒸气蒸馏法提取挥发油,GC-MS测定;GC-MS数据分析与AMDIS软件进行解析.结果:4批陈皮共定性、定量72个化合物,总含量分别为95.542%,92.138%,98.655%,96.201%.含量最高的2个化合物是柠檬烯与芳樟醇,分别占43.78 ~ 68.432%,9.258 ~ 15.724%.结论:4批次陈皮挥发油几乎含有相同的物质成分,各成分在含量上有一定差异.一些物质的量变规律对于揭示陈皮挥发油类成分的动态变化规律具有重要意义.%Objective: To investigate the qualitative and quantitative change law of essential oil components in Citri Reticulatae Pericarpium (CRP) harvested in different time from the same plant. Method: The volatile oils were extracted by steam distillation and analyzed by GC-MS. The overlapped peaks were resolved by the programs of GC-MS data analysis and AMDIS. Result: A total of 72 compounds in the essential oils of four batches CRP were identified, which represented 95.542% , 92. 138% , 98.655% , and 96.201% of the total relative content, respectively. The two compounds that have the highest content are limonene and linalool, accounting for 43.78-68.432% and 9.258-15. 724% respectively. Quantitative changes of some compounds present regularity. Conclusion: The essential oil of each batch CRP has almost the same chemical constituents, whereas the relative content of each constituent shows some difference. The quantitative change law of some components may have important meaning to explore the dynamic change law of volatile oil components in CRP. The combined utilization of programs of GC-MS data analysis and AMDIS can efficiently solve the difficulty in the qualitative and quantitative analysis of overlapped peaks.

  2. Screening Experiments for the Effective Fungicides of Black Rot Disease of Gonggan(Alternaria citri)%贡柑黑腐病有效防治药剂筛选

    Institute of Scientific and Technical Information of China (English)

    邓明学; 谢艳芳; 谭有龙; 覃旭; 覃星瑜; 覃宗典

    2012-01-01

    [目的]筛选防治贡柑黑腐病的有效药剂.[方法]用生长速率法和孢子萌发法测定苯醚甲环唑等15种药剂的EC50、EC95值,并做氟硅唑等5种药剂的田间药效试验.[结果]以EC95(mg a.i./L)排序,抑菌活性较高的有氟硅唑、吡唑醚菌酯·咪鲜胺、吡唑醚菌酯、苯醚甲环唑、咪鲜胺、代森锰锌、丙森锌、百菌清·多菌灵·代森锰锌;田间防效较好的有10%氟硅唑WP、25%苯醚甲环唑·咪鲜胺WP、50%百菌清·多菌灵·代森锰锌WP和72%代森锰锌·霜脲氰WP,而10%氟硅唑EW防效较差.[结论]耐雨水冲刷的杀菌剂和剂型防效较好.%[Aims] This paper aims to screen out the effective fungicides for controlling black rot disease of Gonggan(Alteranria citri). [Methods] The EC50 and EC95(mg a.i./L) of 15 different kinds of fungicides, such as ifenoconazole, pyraclostrobin, were tested with the both methods of the growth rate and spore emergence, meanwhile, the field control of five kinds of fungicides, such as flusilazole, were conducted. [Results] Sorting by EC,,, the effect on inhibition of mycelial growth and spore germination from high to low: flusilazole, pyraclostrobin+prochloraz, pyraclostrobin, difenoconazole, prochloraz, mancozeb, propineb. The field trial results showed that the following .fungicides have good effect: flusilazole 10% WP 1 g/L, difenoconazole+prochloraz 25% WP 1 g/L, chlorothalonil+carbendazim+mancozeb 50% WP 1.67 g/L and mancozeb+cymoxanil 72% WP 1.25 g/L, while flusilazole 10% EW 1 g/L with poor control efficiency. [Conclusions] The control effect is better of the fungicides whose formulations can resist rain erosion.

  3. Rapid characterization of 96 chemical constituents in Citri Reticulatae Folium (leaves of ‘Fuju’) using HPLC-DAD-ESI-MSn%HPLC-DAD-ESI-MSn技术对橘叶(福橘叶)中96个化学成分的快速分离和鉴定

    Institute of Scientific and Technical Information of China (English)

    曹规划; 付庆荣; 张藏曼; 王弘; 陈世忠

    2016-01-01

    为了对橘叶(福橘叶)中的化学成分进行较为系统地研究,我们首次采用液质联用技术((HPLC-DAD-ESI-MSn)系统地分离和鉴定了橘叶中的化学成分.最终,我们在橘叶中解析了96个化学成分,包括:31个酚酸类化合物,4个黄酮苷元,6个黄酮单氧苷,10个黄酮双氧苷,5个黄酮单碳苷和5个黄酮双碳苷化合物,以及6个黄酮碳氧苷,5个3-羟基-3-甲基戊二酰基取代的黄酮苷,1个黄烷-3-醇和2个生物碱类化合物,并且初步鉴定出了21个多甲基黄酮类化合物(PMFs).在上述解析的96个化合物中,52个化合物为首次在橘叶中发现,15个化合物为首次在柑橘属植物中发现.同时,我们在橘叶中也发现了9个潜在的新化合物.%In order to systematically investigate the chemical constituents of Citri Reticulatae Folium (leaves of ‘Fuju’),an analytical method that included high-performance liquid chromatography,diode array detection,electrospray ionization,and ion-trap time-of-flight mass spectrometry (HPLC-DAD-ESI-MSn) was used to separate and identify the individual chemical components of Citri Reticulatae Folium.As a result,96 compounds were tentatively identified in this study:including 31 phenolic acids,4 flavonoid aglycones,6 flavonoid mono-O-glycosides,10 flavonoid-O-diglycosides,5 flavonoid mono-C-glycosides,5 flavonoid di-C-glycosides,6 flavonoid O,C-glycosides,5 (3-hydroxy-3-methylglutaryl) glycosyl flavonoids,1 flavan-3-ol,and 2 alkaloids.In addition,21 polymethoxy flavonoids (PMFs) were identified in this paper.Among these compounds,52 compounds,which were previously found in other Citrus plants,have been identified for the first time in Citri Reticulatae Folium.15 compounds have not been previously found in the Citrus genus were identified.Moreover,9 potentially new compounds have also been detected in this paper.This is the first report of the full characterization of chemical components of Citri Reticulatae Folium (leaves of

  4. Difference analysis of GABA receptor gene between different strains in the red mite, Panonychus citri (Acari: Tetranychidae) of citrus%橘全爪螨不同品系γ氨基丁酸受体基因差异性分析

    Institute of Scientific and Technical Information of China (English)

    张云飞; 李晓娇; 岳建苏; 江高飞; 冉春

    2012-01-01

    【目的】为明确γ氨基丁酸受体基因与橘全爪螨抗性的关系,【方法】通过BLAST搜索,对GABA受体基因进行鉴定:进一步通过序列比对和Sanger测序,对橘全爪螨敏感品系和噻螨酮抗性品系GABA受体基因SNPs进行分析和真实性验证;采用RPKM法对橘全爪螨敏感品系和噻螨酮抗性品系GABA受体基因进行表达差异分析。【结果】从橘全爪螨转录组中获得了19条GABA受体基因。通过敏感品系和抗性品系GABA受体基因序列比较和sanger测序发现Unigene11199_All有2个SNP位点。对GABA受体基因表达差异分析发现,相对于敏感品系,抗性品系中部分GABA受体基因表达量发生了不同程度的变化,抗性品系中有3条GABA受体基因表达上调,13条GABA受体基因表达下调,Unigene24440_All下调倍数最高[10g2Ratio(RS/SS)=-10.452479]。【结论】由此推断,橘全爪螨抗性产生可能是一个比较复杂的过程,而Unigenelll99_All和Unigene24440_All可能是橘全爪螨对噻螨酮产生抗性的重要原因。%[Objective]In order to clarify the relationship between T-aminobutyric acid receptor gene and resistance of Panonychus citri. [Method]GABA receptor genes were identified by BLAST searching. SNPs in GABA receptor genes between susceptible strain and hexythiazox-resistant strain of P. citri were ana- lyzed and examined by comparison of GABA receptor gene sequences and Sanger sequencing, and gene expression profiles of GABA receptor genes were further compared using RPKM (Reads Per kb per Million reads) method. [Result] 19 GABA receptor genes were found in transcripts of P. citri. Comparisons of GA- BA receptor gene sequences and further sequencing showed that there were 2 SNPs in Unigenel 1199_All between different strains. Analysis of difference of gene expression showed 3 GABA receptor genes were up-regulated and 13 were down-regulated, of which, Unigenel1069_All was the top down

  5. 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.

    Science.gov (United States)

    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.

  6. 福建省柑橘黄龙病危害及其流行规律研究%Damage and Epidemics of Citrus Huanglongbing in Fujian Province

    Institute of Scientific and Technical Information of China (English)

    谢钟琛; 李健; 施清; 谢文龙; 杨建榕; 陈跃飞; 钟连生

    2009-01-01

    [目的]全面了解柑橘黄龙病(Citrus Huanglongbing,简称HLB)在福建省的危害分布及其流行情况,有效指导HLB的防控.[方法]对全省48个县(市、区)及245个柑橘主产乡镇HLB虫媒柑橘木虱(Diaphorina citri)分布及HLB的危害进行调查,以PCR(polymerase chain reaction,PCR)方法对采集的斑驳黄化柑橘叶片、红鼻果进行确证性检测,分析HLB的危害流行规律,并应用滑动相关分析筛选影响HLB流行的气象要素.[结果]全省各县及柑橘主产乡镇均可查到柑橘木虱和HLB病株(最北N28°07'.海拔最高880 m),HLB危害程度为平均发病率16.59%,主栽品种间感病性差异显著,排序为宽皮桔>甜橙类>柚类;HLB发病率与果园纬度、海拔、坡度呈显著负相关;在南亚热带气候区的长泰县(N24°07'),HLB田间发病率仅与临近2年春季气候有关,推测HLB潜伏期为6~18个月,同时春梢期日照、幼果期湿度与HLB发病率呈极显著正、负相关,"春旱"为HLB流行性气候;柑橘红鼻果、斑驳黄化叶片的HLB病原Nested-PCR检出率分别为93.8%、73.1%,前者可靠性优于后者,二类症状均可视为HLB的典型症状.[结论]福建境内的柑橘产区均有HLB病原与柑橘木虱的分布.若春季遇严重干旱HLB就有局部爆发的可能.柑橘红鼻果可作HLB田间诊断依据.%[Objective] To effectively control citrus huanglongbing (HLB, yellow shoot disease), the distribution, damage and epidemics of the disease were investigated in Fujian Province. [Method] The distribution of"Candidatus Liberibacter asiaticus", the putative pathogen, the Asian citrus psyllid (Diaphorina citri), the insect vectors, and the incidence of HLB were investigated in 245 citrus-producing towns in 48 counties in Fujian Province. Polymerase chain reaction (PCR) assay was carried out to verify the existence of HLB in the collected blotchy mottle leaves and color inversion fruits to analyze the epidemics of HLB. Sliding correlation

  7. Laser system for identification, tracking, and control of flying insects.

    Science.gov (United States)

    Mullen, Emma R; Rutschman, Phillip; Pegram, Nathan; Patt, Joseph M; Adamczyk, John J; Johanson, 3ric

    2016-05-30

    Flying insects are common vectors for transmission of pathogens and inflict significant harm to humans and agricultural production in many parts of the world. We present proof of principle for an optical system capable of highly specific vector control. This system utilizes a combination of optical sources, detectors, and sophisticated software to search, detect, and identify flying insects in real-time, with the capability of eradication using a lethal laser pulse. We present data on two insect species to show species distinction; Diaphorina citri, a vector of the causal agent of citrus greening disease, and Anopheles stephensi, a malaria vector.

  8. 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 ...

  9. HPLC Determination of Naringin, Hesperidin and Neohesperidin in Citri Reticulatae Pericarpium and Aurantii Fructus Extract%HPLC测定陈皮-枳壳药对提取物中柚皮苷、橙皮苷和新橙皮苷的含量

    Institute of Scientific and Technical Information of China (English)

    郭琦丽; 吕武清; 曾莉萍; 李才堂

    2012-01-01

    目的:建立陈皮-枳壳药对提取物中柚皮苷、橙皮苷和新橙皮苷的含量测定方法.方法:采用HPLC,Agilent Hypersill C18(4.0 mm×250 mm,5μm)色谱柱,以乙腈-0.1%磷酸(24∶ 76)为流动相,流速1 mL· min-1,检测波长283 nm,柱温40℃.结果:柚皮苷、橙皮苷和新橙皮分别在6.05 ~60.5,4.34 ~ 43.4,7.14 ~71.40 mg·L-1线性关系良好(r均>0.999 5);加样回收率分别为98.65%,98.66%,100.78%,其RSD分别为2.01%,1.52%,2.11%;重复性RSD分别为2.41%,1.56%,2.52%(n=6).结论:该方法简便可行,重复性好,结果准确可靠,可用于陈皮-枳壳药对提取物中柚皮苷、橙皮苷和新橙皮苷的含量测定.%Objective: To develop a method to determine naringin, hesperidin and neohesperidin in Citri Reticulatae Pericarpium and Aurantii Fructus. Method; HPLC was used to determine directly naringin, hesperidin and neohesperidin of Citri Reticulatae Pericarpium and Aurantii Fructus. Phenomsil C18 (4. 0 mm x 250 mm, 5 μm) as the chromatographic column, and acetonitrile-10% aqueous phosphoric acid (24:76) as the mobile phase, the flow rate of 1.0 mL -min , the column temperature was at 40 °C. Result; The linearity of verbascoside was good in the range of 6. 05-60. 5 , 4. 3443. 4 and 7. 14-71. 40 mg - L-1 ( r > 0. 999 5 ) . The recovery of the added sample was 98. 65% , 98. 66% and 100. 78% with RSD of 2. 01% , 1. 52% and 2. 11% . Conclusion.- The method is simple, accurate and can be used for the determination of naringin, hesperidin and neohesperidin in Tangerine peel and Aurantii Fructus extract.

  10. 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

  11. 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

  12. Wolbachia-infection differs among potato psyllid haplotypes

    Science.gov (United States)

    Wolbachia is a bacterial endosymbiont of insects that can manipulate insect reproduction. In many insects, Wolbachia-free females cannot produce viable offspring when mated by infected males. The manipulation of insect reproduction by Wolbachia has important implications for insect evolution and pop...

  13. Antifungal activity of chitosan(oligochitosan) on Geotrichum candidum and Alternaria citri of citrus and postharvest disease control%壳聚(寡)糖对柑橘酸腐、黑腐病菌的抑制作用及对采后病害的防治

    Institute of Scientific and Technical Information of China (English)

    王丽; 赵盼; 孟祥红

    2011-01-01

    In vitro,the pore germination,germ tube length and mycelial growth of the two fungi were used to evaluate to the effect of chitosan and oligochitosan. The effects of chitosan and ologochitosan on disease control were investigated by stab inoculation. The results showed that chitosan and oligochitosan at 10mg/mL almost completely inhibited the mycelial growth of the two fungi. Chitosan and oligochitosan at 0. 5mg/mL completely inhibited the spore germination and germ tube elongation of Geotrichum candidum. Moreover,2mg/mL of chitosan could completely inhibit the spore germination and germ tube elongation of Alternaria citri,but 2mg/mL of oligochitosan had a lower inhibitory effect. In vivo results showed that the control effect of chitosan and oligochitosan at 1% on fruit disease were better than lower concentration.%通过体外实验测定不同浓度的壳聚糖和壳寡糖对酸腐和黑腐病菌孢子萌发、芽管长度和菌丝生长的抑制作用;采用刺伤接种方法评价壳聚糖和壳寡糖对柑橘果实两种病害的防治作用。离体实验结果表明,壳聚糖和壳寡糖浓度为10mg/mL时几乎完全抑制两种病原菌的菌丝生长;浓度为0.5mg/mL壳聚糖和壳寡糖能完全抑制柑橘酸腐病菌(Geotrichum candidum)孢子萌发和芽管伸长的效果,而2mg/mL的壳聚糖才能完全抑制柑橘黑腐病菌(Alternaria citri)孢子萌发和芽管伸长,但相同浓度的壳寡糖抑制A.citri的孢子萌发和芽管伸长效果较差。体内实验结果表明,其病害防治效果具有浓度依赖性,高浓度(1%)的壳聚糖和壳寡糖对果实病害的防治效果较低浓度的防治效果好。

  14. Development of RLK-Derived Molecular Markers Associating with the Resistance to Citrus Canker [Xanthomonas axonopodis pv. Citri (Xac)] Disease%开发与柑桔抗溃疡病基因连锁的由RLK-RGC衍生的分子标记

    Institute of Scientific and Technical Information of China (English)

    向旭; 郑启发; 黄舒; 陈存贤; Frederick G. Gmitter Jr.; 邓占鳌

    2005-01-01

    本文是在先期研究工作的基础上,利用从柑桔与枳属间杂种的基因组DNA所获得的类受体激酶(RLK)的候选抗病基因序列,重新设计引物,对柑桔抗溃疡病材料和感病材料开展以PCR扩增为基础的对比分析,其中一对引物‘19h16/DdeI'的扩增产物经限制性内切酶DdeI酶切,揭示了抗性材料(Ichang Papeda,Meiwa kumquat,Maruhi kumquat and Nagami kumquat)和感病材料(Flying Dragon,Valencia orange and Palestine sweetlime)之间的多态性差异,同时,在C.Ichangensis的自交F1代和Palestine sweetlime× Ichang Papeda的杂交F1代群体中也见明显的多态性差异;对这些F1代个体进行溃疡病病原[Xanthomonas axonopodis pv.Citri(Xac)]接种作抗病性表型鉴定,结果表明,该分子标记‘19h16/DdeI'与柑桔溃疡病抗性密切相关;经纯化测序的结果进一步证明,抗性材料PCR扩增产物具有完全一致的序列和DdeI酶切位点,但感病材料缺少这一DdeI酶切位点;染色体步行(primerwalking)在BAC文库的对应单克隆中获得了一个完整的抗病基因序列,与以前获得的2个抗病基因序列‘17o6RLKP'和‘26m19RLKP'相比,‘19h16RLKP'也具有Xa21抗病基因蛋白的所有特征,包括含有一个信号肽、同样数目的亮氨酸重复序列、跨膜域和激酶域等.基于该序列的开放读码框,发展了特异性更强可靠性更高的另一分子标记,在今后的研究工作中具有较大的应用潜力.%By utilizing the receptor-like kinase (RLK) RGC sequences previously obtained from the genomic DNA of an intergeneric Citrus and Poncirus hybrid, PCR amplification with one pair of new-designed primers was found to reveal polymorphism between the theoretical resistant species (Ichang Papeda, Meiwa kumquat,Maruhi kumquat and Nagami kumquat) and susceptible species (Flying Dragon, Valencia orange and Palestine sweet lime ) and among the self-crossing population of C. Ichangensis and F1 population of Palestine sweet lime

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  17. 药剂防控柑橘黄龙病的探讨%Discussion on Pesticides Controlling the Citrus Huanglongbing

    Institute of Scientific and Technical Information of China (English)

    成家壮

    2011-01-01

    Based on the characteristics and the epidemic law of citrus disease,an idea was put forwardresearching and developing the antifeedant against diaphorina citri and the bactericides to control pathogenic bacteria andto bukme the host plant resistance in order to control Huanglongbing(HLB),at the same time,the relevant issues were alsodiscussed.%根据柑橘黄龙病的发生特点与流行规律,提出了研究开发柑橘木虱拒食剂和抑制病原菌以及可诱导寄主抗病性的杀菌剂用以防控柑橘黄龙病的构想,并对相关问题进行了探讨.

  18. Monitoring and controlling measures of Huanglongbin disease in Guanxi pomelo plant%琯溪蜜柚黄龙病的监测与防控

    Institute of Scientific and Technical Information of China (English)

    杨文火

    2012-01-01

    通过对采集的琯溪蜜柚的各种典型黄花症状样品进行黄龙病病原PCR测定,在分析病害症状并进行实时监测的基础上,提出及时铲除病株和防治木虱、严格控制外来种苗的进入等防控措施。%Pathogen of Citrus Huanglongbing(HLB)was detected by PCR techque by using the samples of Guanxi pomelo,which showed typical leaf-yellowing symptom.Based on analyzing the symptom of HLB and on-the-spot monitoring to the disease,we put forward a set of integrated control techniques,including removing HLB-affected plants,controlling Diaphorina citri,strictly forbiding the entrance of alien seedings.

  19. 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.

  20. Invasive mutualisms between a plant pathogen and insect vectors in the Middle East and Brazil.

    Science.gov (United States)

    Queiroz, Renan Batista; Donkersley, Philip; Silva, Fábio Nascimento; Al-Mahmmoli, Issa Hashil; Al-Sadi, Abdullah Mohammed; Carvalho, Claudine Márcia; Elliot, Simon L

    2016-12-01

    Complex multi-trophic interactions in vectorborne diseases limit our understanding and ability to predict outbreaks. Arthropod-vectored pathogens are especially problematic, with the potential for novel interspecific interactions during invasions. Variations and novelties in plant-arthropod-pathogen triumvirates present significant threats to global food security. We examined aspects of a phytoplasma pathogen of citrus across two continents. 'Candidatus Phytoplasma aurantifolia' causes Witches' Broom Disease of Lime (WBDL) and has devastated citrus production in the Middle East. A variant of this phytoplasma currently displays asymptomatic or 'silent' infections in Brazil. We first studied vector capacity and fitness impacts of the pathogen on its vectors. The potential for co-occurring weed species to act as pathogen reservoirs was analysed and key transmission periods in the year were also studied. We demonstrate that two invasive hemipteran insects-Diaphorina citri and Hishimonus phycitis-can vector the phytoplasma. Feeding on phytoplasma-infected hosts greatly increased reproduction of its invasive vector D. citri both in Oman and Brazil; suggesting that increased fitness of invasive insect vectors thereby further increases the pathogen's capacity to spread. Based on our findings, this is a robust system for studying the effects of invasions on vectorborne diseases and highlights concerns about its spread to warmer, drier regions of Brazil.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  3. The community composition and temporal dynamic of canopy arthropods on Citrus in Nanchang, Jiangxi Province%江西南昌柑橘冠层节肢动物群落组成及时序动态研究

    Institute of Scientific and Technical Information of China (English)

    李小珍; 王建国; 肖海军; 刘伟; 鲍涵涵; 廖琪

    2013-01-01

    采用叶面观察和诱捕相结合的方法调查了江西南昌柑橘冠层节肢动物群落.共调查到节肢动物数量144037头,分属25种.其中,高密度种类有2种即柑橘全爪螨和矢尖蚧;常见种类有13种,包括桔小实蝇、柑橘花蕾蛆、柑橘木虱和中华草蛉等;其它为低密度种类.在群落结构指数的变化过程中,分别于5月下旬和9月下旬出现Shannon-Wiener多样性指数极大值和Simpson优势度指数极小值.对高密度种类和部分常见种类的种群动态进行了分析,表明柑橘全爪螨在5月下旬至6月上旬发生严重,矢尖蚧则于7~8月发生严重;柑橘花蕾蛆幼虫和柑橘木虱发生动态相似,均仅于柑橘春梢阶段出现最高密度;桔小实蝇成虫则在9月下旬出现密度高峰;中华草蛉的发生动态与其主要捕食对象一柑橘全爪螨的相似.研究结果初步明确南昌柑橘冠层节肢动物的重要成分及重要类群的时序动态,为该地区柑橘重要害虫的防治与控制,天敌昆虫的保护与利用提供基础信息.%The composition and structure of the arthropod communities in orange orchard were investigated in Nanchang,Jiangxi Province,using the leaf observational method and the trapping technique from April to December 2012.A total of 144037 individuals,belonging to 25 species,were founded in our investigation.Among them,the high density species included with Panonychus citri MeGregor and Unaspis yanonensis (Kuwana),13 kinds of common species including Diaphorina Citri (Kuwayama),Bactrocera dorsalis Hendel,Contarinia citri Barnes,Chrysoperla sinica (Tjeder) and so on,and others were the low density species.During the change of arthropod community indexes,Shannon -Wiener diversity index reached their maximum values,and Simpson dominance index appeared their minimum values in late May and late September,respectively.Subsequently,we described the population dynamics of high density species and some common species

  4. Repellency of selected biorational insecticides to potato psyllid, Bactericera cockerelli (Hemiptera: Psyllidae)

    Science.gov (United States)

    Bactericera cockerelli has recently become a major concern because of its direct feeding and vectoring of bacterial diseases in many solanaceous crops. The repellency of four biorational insecticides, MOI-201 (a Chinese medicine plant extract), Requiem (a plant extract of Chenopodium ambrosioides), ...

  5. 75 FR 34322 - Citrus Greening and Asian Citrus Psyllid; Quarantine and Interstate Movement Regulations

    Science.gov (United States)

    2010-06-17

    ...) leaves that are intended for culinary use are usually both dried and shredded prior to shipment. After.../report_pest_disease/report_pest_disease.shtml ). After a change is made to the list of quarantined...

  6. 77 FR 59709 - Citrus Greening and Asian Citrus Psyllid; Quarantine and Interstate Movement Regulations

    Science.gov (United States)

    2012-10-01

    ...: Effective October 31, 2012. FOR FURTHER INFORMATION CONTACT: Ms. Lynn Evans-Goldner, National Program... Huanglongbing disease of citrus by the international taxonomic community. We are amending the definition to... issued under Sec. 204, Title II, Public Law 106-113, 113 Stat. 1501A-293; sections 301.75-15 and...

  7. Economic injury level of the psyllid, Agonoscena pistaciae, on Pistachio, Pistacia vera cv. Ohadi.

    Science.gov (United States)

    Reza Hassani, Mohammad; Nouri-Ganbalani, Gadir; Izadi, Hamzeh; Shojai, Mahmoud; Basirat, Mehdi

    2009-01-01

    The pistachio psylla, Agonoscena pistaciae Burckhardt and Lauterer (Hemiptera: Psyllidae) is a major pest of pistachio trees, Pistacia vera L. (Sapindalis: Anacardiaceae) throughout pistachio-producing regions in Iran. Different density levels of A. pistaciae nymphs were maintained on pistachio trees by different insecticide dosages to evaluate the relationship between nymph density and yield loss (weight of 1000 nuts). Psylla nymph densities were monitored weekly by counting nymphs on pistachio terminal leaflets. There was a significant reduction in weight of 1000 nuts as seasonal averages of nymphs increased. Regression analysis was used to determine the relationship between nymph density and weight of 1000 nuts. The economic injury levels varied as a function of market values, management costs, insecticide efficiency and yield loss rate and ranged from 7.7 to 30.7 nymphal days per terminal leaflet, based on weight of 1000 nuts.

  8. Field ID guide to citrus relative hosts of Asian citrus psyllid & Huanglongbing

    Science.gov (United States)

    The Rutaceae family of plants includes not only species within the genus Citrus, but also several other genera and species that may not be easily recognized as having any relationship to citrus at all. However, many of these citrus relatives are used for ornamental, culinary, or religious purposes. ...

  9. 76 FR 23449 - Citrus Canker, Citrus Greening, and Asian Citrus Psyllid; Interstate Movement of Regulated...

    Science.gov (United States)

    2011-04-27

    ... analysis, especially any scientific studies that may be pertinent. Protocol Document Based on the... either case, the statement is prominently and legibly displayed. The nursery stock is moved in a... Huanglongbing. Plant Management Network International, Orlando, Florida. Copies available from the individuals...

  10. Anoeconeossa bundoorensis sp. n., a new psyllid (Hemiptera: Psylloidea) from Eucalyptus camaldulensis (Myrtaceae) from Southeast Australia.

    Science.gov (United States)

    Taylor, Gary S; Farnier, Kevin; Burckhardt, Daniel; Steinbauer, Martin J

    2013-01-30

    Anoeconeossa bundoorensis sp. n. is described from Eucalyptus camaldulensis (Myrtaceae) from southern Victoria in Southeast Australia. It is placed in the A. communis Taylor species-group as the paramere lacks combs of black rods. It differs from other members of the species-group, A. communis and A. bullata Taylor as it lacks an anterobasal expansion on the paramere and from A. unicornuta Taylor as the inner horn-shaped process of the paramere is reduced to a short spine and the apical expansion is more elongate, with a corresponding greater length of equidistant setae. Taxonomically relevant morphological details are illustrated and the species is diagnosed from other eucalypt inhabiting congeners. In-formation on the biology is presented.

  11. Use of psyllid genomes RNA interference for novel pest control strategies

    Science.gov (United States)

    Genomics has changed the strategies used to manage insects and diseases. The ability to effect a change in proteins, and transcripts, through RNA-interference, RNAi, has produced a rush towards the development of the most state-of-the-art pest suppression strategies available. To rapidly advance the...

  12. 78 FR 63369 - Citrus Canker, Citrus Greening, and Asian Citrus Psyllid; Interstate Movement of Regulated...

    Science.gov (United States)

    2013-10-24

    ... infected plants, which render the fruit unmarketable, and cause infected fruit to drop from the trees... of the fruit, and can kill trees. Once infected, there is no cure for a tree with citrus greening. In... disease that is caused by a complex of Xanthomonas spp. bacteria and that affects plants and plant...

  13. Bacteria of the genus Erwinia found in the spermatheca of the laurel psyllid Trioza alacris.

    Science.gov (United States)

    Marchini, Daniela; Ciolfi, Silvia; Gottardo, Marco; Marri, Laura

    2014-12-01

    Psylloidea are economically important insects causing serious damage to plants by direct feeding and/or vectoring bacterial pathogens. Results reported here indicate the presence of extracellular bacteria in the spermatheca of egg-laying Trioza alacris females. One phylotype, sharing 99 % identity with the non-phytopathogenic bacterium Erwinia tasmaniensis, was identified regardless of methods applied or insect sampling year and location. This is the first study, achieved by ultrastructural, cultural, and 16S rRNA gene-based analysis, of an insect spermatheca microbiota.

  14. 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 semi

  15. Temporal variation in sex allocation in the mealybug Planococcus citri : Adaptation, constraint, or both?

    NARCIS (Netherlands)

    Ross, Laura; Langenhof, Minke B. W.; Pen, Ido; Shuker, David M.

    2012-01-01

    Sex ratio theory has been very successful in predicting under which circumstances parents should bias their investment towards a particular offspring sex. However, most examples of adaptive sex ratio bias come from species with well-defined mating systems and sex determining mechanisms, while in man

  16. Parallel evolution of cytochrome b mediated bifenazate resistance in the citrus red mite Panonychus citri

    NARCIS (Netherlands)

    Van Leeuwen, T.; Van Nieuwenhuyse, P.; Vanholme, B.; Dermauw, W.; Nauen, R.; Tirry, L.

    2011-01-01

    Bifenazate is a recently developed acaricide that is mainly used to control spider mites on a variety of crops. Although first thought to be a neurotoxin, genetic evidence obtained from bifenazate resistant Tetranychus urticae strains suggested an alternative mode of action as a Qo pocket inhibitor

  17. Molecular evidence of polyandry in the citrus mealybug, Planococcus citri (Hemiptera: Pseudococcidae.

    Directory of Open Access Journals (Sweden)

    Sofia G Seabra

    Full Text Available The occurrence of polyandry in Planococcuscitri, presumed by earlier observations of mating behavior, was confirmed using microsatellite genotyping of pools of over 400 eggs resulting from controlled crosses of one female with two males. The genetic contribution of both mated males was confirmed in 13 out of 43 crosses. In three crosses it was possible to determine that only the first male fertilized the eggs, which may be due to sperm competition or unviable sperm supply. The microsatellite analysis also allowed the confirmation of aspects of the chromosomal inheritance detected previously in cytogenetic studies in Planococcuscitri, namely that only one of the alleles is transmitted by the male, indicating that the males are functionally haploid, supporting the observation of Paternal Genome Elimination (PGE in these insects.

  18. Molecular evidence of polyandry in the citrus mealybug, Planococcus citri (Hemiptera: Pseudococcidae).

    Science.gov (United States)

    Seabra, Sofia G; Brás, Patricia G; Zina, Vera; Borges da Silva, Elsa; Rebelo, Maria Teresa; Figueiredo, Elisabete; Mendel, Zvi; Paulo, Octávio S; Franco, José Carlos

    2013-01-01

    The occurrence of polyandry in Planococcuscitri, presumed by earlier observations of mating behavior, was confirmed using microsatellite genotyping of pools of over 400 eggs resulting from controlled crosses of one female with two males. The genetic contribution of both mated males was confirmed in 13 out of 43 crosses. In three crosses it was possible to determine that only the first male fertilized the eggs, which may be due to sperm competition or unviable sperm supply. The microsatellite analysis also allowed the confirmation of aspects of the chromosomal inheritance detected previously in cytogenetic studies in Planococcuscitri, namely that only one of the alleles is transmitted by the male, indicating that the males are functionally haploid, supporting the observation of Paternal Genome Elimination (PGE) in these insects.

  19. 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

    National Research Council Canada - National Science Library

    María E. Romero; C. Jacqueline Ramallo; L. Daniel Ploper

    2008-01-01

    ... 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...

  20. 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

    OpenAIRE

    María E. Romero; C. Jacqueline Ramallo; L. Daniel Ploper

    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) ...

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  3. 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...

  4. Analysis of Transcriptome Differences between Resistant and Susceptible Strains of the Citrus Red Mite Panonychus citri (Acari: Tetranychidae)

    OpenAIRE

    Bin Liu; Gaofei Jiang; Yunfei Zhang; Junli Li; Xiaojiao Li; Jiansu Yue; Fei Chen; Haoqiang Liu; Hongjun Li; Shiping Zhu; Jinjun Wang; Chun Ran

    2011-01-01

    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...

  5. 台州柑橘黄龙病防控研究新进展%The Control Research Progress of Liberobacter asiaticum in Taizhou City

    Institute of Scientific and Technical Information of China (English)

    余继华; 张敏荣; 陶健; 卢璐; 杨晓; 汪恩国; 叶志勇

    2015-01-01

    根据十几年对柑橘木虱和柑橘黄龙病发生规律与防控技术研究探讨,明确了柑橘木虱在台州橘林的发生分布规律和抽样技术,分析了柑橘木虱虫株率和带毒率与黄龙病、黄龙病与柑橘产量损失等之间的关系,探讨了柑橘黄龙病预警和柑橘木虱防治指标,建立了不同管理方式下的疫情扩散模型预测橘林经济寿命,提出了柑橘黄龙病综合防控技术和可防可控的理念。%After decades of researches and discussions on occurrence regularity and control technology of citrus psyl id (Diaphorina citri Kuwayana) and Liberobacter asi-aticum, the occurrence regularity and sampling technique of citrus psyl id in orange forests in Taizhou was defined and the connections between the infection rate and carrying rate of citrus psyl id and Liberobacter asiaticum as wel as the correlations between the Liberobacter asiaticum and citrus yield loss were discussed. This paper discussed the warnings of Liberobacter asiaticum and citrus psylid control index, the economic life span of orange forests was then predicted by building the diseases spreading models with different management styles. At last, the paper put forward the comprehensive prevention and control technology and the concept that Liber-obacter asiaticum was preventable and control able.

  6. ISSN 1816-0573 CAFOBIOS

    African Journals Online (AJOL)

    TEG

    Phenology and fruit production of Piliostigma reticulatum (DC), Hochst., an agroforestry forage species in the Sahel. ... [3]; Citrus psyllid, Trioza erytreae Del Guercio. 1918 [7]; ..... Tuthill, L.D. 1943: The Psyllids of America north of Mexico ...

  7. 76 FR 4859 - Availability of an Environmental Assessment and Finding of No Significant Impact for a Biological...

    Science.gov (United States)

    2011-01-27

    ... Significant Impact for a Biological Control Agent for Asian Citrus Psyllid AGENCY: Animal and Plant Health... continental United States for use as a biological control agent to reduce the severity of Asian citrus psyllid... use as a biological control agent to reduce the severity of Asian citrus psyllid (ACP)...

  8. [First record of Zoophthora radicans (Entomophthorales: Entomophthoraceae) on adults of the Paraguay tea psyllid, Gyropsylla spegazziniana Lizer & Trelles (Hemiptera: Psyllidae), in Brazil].

    Science.gov (United States)

    Alves, Luis F A; Leite, Luis G; Oliveira, Daian G P de

    2009-01-01

    We report the first occurrence of Zoophthora radicans infecting adults of Gyropsylla spegazziniana Lizer & Trelles in a commercial Paraguay tea plantation (Ilex paraguariensis), in Cascavel, PR, Brazil. The fungus prevalence was high (90% of mortality), considered a natural epizooty.

  9. A draft whole genome sequence of “Candidatus Liberibacter asiaticus” strain TX2351 from Asian citrus psyllids in Texas, USA

    Science.gov (United States)

    The draft genome sequence of “Candidatus Liberibacter asiaticus” strain TX2351 collected from ACP in South Texas has been determined. The TX2351 genome is 1,252,043 bp in size with a 36.5% G+C content, encoding 1,184 predicted open reading frames and 51 RNA genes....

  10. Study on HPLC Fingerprint of Folium Citri Grandis of Tongxian%通贤柚叶HPLC指纹图谱研究

    Institute of Scientific and Technical Information of China (English)

    游元元; 范媛媛; 陈席; 谢仁宗

    2014-01-01

    目的 建立通贤柚叶的HPLC指纹图谱.方法 色谱柱为Sepax Amethyst C18-H(250 mm×4.6 0mm,5μm),流动相为乙腈-0.1%甲酸,梯度洗脱,流速1.0 mL· min-1,检测波长303 nm,柱温25℃,进样量20μL.分析12批通贤柚叶样品,并进行主成分分析.结果 建立了通贤柚叶的HPLC指纹图谱,确定了16个共有色谱峰,采用对照品指认了其中2个峰.主成分分析结果表明,11个色谱峰对其质量控制较为重要.结论 所建立的指纹图谱分析方法特征性强,为通贤柚叶的质量控制提供了科学依据.

  11. The occurrence, prevention and cure of Exocarpium Citri Grandis plant diseases and insect pests%化橘红病虫害的发生与防治

    Institute of Scientific and Technical Information of China (English)

    蔡岳文; 严振; 丘金裕

    2004-01-01

    化橘红为芸香科植物化州柚(Citrus grandis(L)Osbeck var.tomentosa Hort)以未成熟果实的外果皮入药。近几年来,随着基地化、规模化大面积种植,其病虫害的发生及危害程度亦与日俱增,一些重要害虫已严重影响化橘红的生产和产品质量。

  12. 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.

    2010-01-01

    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 thei

  13. Tale of the Huanglongbing Disease Pyramid in the Context of the Citrus Microbiome.

    Science.gov (United States)

    Wang, Nian; Stelinski, Lukasz L; Pelz-Stelinski, Kirsten S; Graham, James H; Zhang, Yunzeng

    2017-04-01

    The Huanglongbing (HLB) disease pyramid is composed of Liberibacters, psyllid vectors, citrus hosts, and the environment. The epidemiological outcomes for Liberibacter-associated plant diseases are collectively determined by the inherent relationships among plant-Liberibacters-psyllids, and how various environmental factors affect plant-Liberibacter-psyllid interactions. Citrus-Liberibacter-psyllid interactions occur in a complex microbiome system. In this review, we focus on the progress in understanding the HLB disease pyramid, and how the microbiome affects the HLB disease pyramid including the interaction between HLB and the citrus microbiome; the interaction between Liberibacters and psyllids; the interaction between Liberibacters and gut microbiota in psyllids; and the effect of HLB on selected above- and belowground citrus pathogens. Their implications for HLB management are also discussed.

  14. Solanum habrochaites, a potential source of resistance against Bactericera cockerelli (Hemiptera: Triozidae) and "Candidatus Liberibacter solanacearum".

    Science.gov (United States)

    Levy, Julien; Tamborindeguy, Cecilia

    2014-06-01

    The potato psyllid, Bactericera cockerelli Sulc, also known as tomato psyllid, is a serious pest of solanaceous plants. Its host selection criteria are poorly understood. In this study, we tested whether the Solanum habrochaites (PI127826), a wild solanaceous plant known for its property to repel whiteflies, was repellent to potato psyllids. Using a combination of nonchoice assays and choice assays on different psyllid stages, we demonstrated that S. habrochaites is both repelling and toxic to potato psyllids compared with Solanum lycopersicum. However, those properties were not sufficient to avoid. transmission of the plant bacterial pathogen "Candidatus Liberibacter solanacearum" vectored by potato psyllids, the causative agent of potato zebra chip disease. However, a lower bacterial transmission rate to S. habrochaites was observed compared with S. lycopersicum.

  15. Micro computed tomography (CT) scanned anatomical gateway to insect pest bioinformatics

    Science.gov (United States)

    An international collaboration to establish an interactive Digital Video Library for a Systems Biology Approach to study the Asian citrus Psyllid and psyllid genomics/proteomics interactions is demonstrated. Advances in micro-CT, digital computed tomography (CT) scan uses X-rays to make detailed pic...

  16. Ecology of the Asian citrus pysllid

    Science.gov (United States)

    Host selection by psyllids in general appears to involve taste rather than olfaction. Adults are often less discriminating than nymphs. A priori, there is good reason to doubt that Asian citrus psyllid (ACP) uses a long-distance sex pheromone or that ACP orients to host plant volatiles over large (m...

  17. Identification of Bacterial Plant Pathogens Using Multilocus Polymerase Chain Reaction/Electrospray Ionization-Mass Spectrometry

    Science.gov (United States)

    2008-01-01

    seedlings (sweet orange ) were inoculated with X. citri subsp. citri (also known as X. axonopodis pv. citri). Cabbage plants were inoculated with X...caryophylli PC113; ATCC25418 Carnation ; USA 1 Burkholderia gladioli pv. gladioli FC-368; PM107; ATCC10248 Gladiolus; USA 1 Clavibacter michiganensis...alfalfae pv. citrumelosis No call – Citrus inoculated with Xanthomonas alfalfae pv. citrumelosis No call – Healthy citrus (sweet orange ) No call

  18. 宜昌市柑桔病虫发生新动态及防治策略%Dynamics and control strategy of citrus pests in Yichang City

    Institute of Scientific and Technical Information of China (English)

    张国宝

    2000-01-01

    本文着重介绍了近几年来,宜昌市柑桔红蜘蛛(Panxonychus citri McGregor)、矢尖蚧(Unaspis yanonensis Kuwana)、蚜虫(Aphis citricidus Kirkaldy)、大实蝇(Tetradacus citri Chen)、吉丁虫(Agrilus citri Matsumura)、疮痂病(Elsinoe fawcetti But.et Jenk)、煤烟病(Capncdium citri Berk.et Desm)等发生新动态,通过研究分析其新动态,针对性地提出了今后柑桔病虫的综防策略.

  19. Transcriptome analyses of Bactericera cockerelli adults in response to "Candidatus Liberibacter solanacearum" infection.

    Science.gov (United States)

    Nachappa, Punya; Levy, Julien; Tamborindeguy, Cecilia

    2012-10-01

    The potato/tomato psyllid, Bactericera cockerelli (Šulc) is an economically important crop pest that not only causes damage through its feeding but also transmits the bacterium, "Candidatus Liberibacter solanacearum" (CLs), which causes zebra chip disease in potato. There is some information about the phenotypic effects of phytopathogenic bacteria on their insect vectors; however, there are no published reports of the molecular mechanisms underlying phytopathogenic bacteria-insect vector interaction. In order to investigate the effects of CLs infection on B. cockerelli, transcriptomic analyses of CLs-infected and uninfected adult psyllids that were reared on potato were performed. De novo assembly of cDNA sequences generated 136,518 and 109,983 contigs for infected and uninfected insect libraries with an average contig length of 514 bp. BlastX analysis against the NCBI-nr database revealed that 33.33 % had significant matches. Gene ontology data illustrated that the majority of the expressed psyllid genes are involved in metabolic process, biological regulation, binding and catalytic activity. The psyllid transcriptome had an abundance of genes such as vitellogenin, heat shock protein, ejaculatory bulb-specific protein, ferritin, and cytochrome oxidase. Notably absent in the psyllid transcriptome were innate immunity genes induced in response to Gram-negative bacteria (IMD pathway). Several functionally diverse contigs related to symbiotic bacteria including the primary endosymbiont Carsonella ruddii, Wolbachia, and CLs in the psyllid transcriptome were identified. A total of 247 contigs showed differential expression in response to CLs infection including immune and stress-related genes and vitellogenins. Expression analyses of selected psyllid genes were performed on psyllids that were exclusively reared on potato (host of the insects used for RNAseq) and psyllids exclusively reared on tomato (alternative host of psyllids). These genes showed similar expression

  20. NCBI nr-aa BLAST: CBRC-XTRO-01-2352 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-2352 ref|NP_642468.1| YapH protein [Xanthomonas axonopodis pv. citri s...tr. 306] gb|AAM37004.1| YapH protein [Xanthomonas axonopodis pv. citri str. 306] NP_642468.1 0.30 32% ...

  1. NCBI nr-aa BLAST: CBRC-LAFR-01-1946 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-1946 ref|NP_641456.1| hypothetical protein XAC1119 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM35992.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_641456.1 0.50 39% ...

  2. NCBI nr-aa BLAST: CBRC-XTRO-01-2343 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-2343 ref|NP_642141.1| filamentous haemagglutinin [Xanthomonas axonopod...is pv. citri str. 306] gb|AAM36677.1| filamentous haemagglutinin [Xanthomonas axonopodis pv. citri str. 306] NP_642141.1 6e-04 27% ...

  3. NCBI nr-aa BLAST: CBRC-LAFR-01-0106 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-0106 ref|NP_643310.1| MFS transporter [Xanthomonas axonopodis pv. citr...i str. 306] gb|AAM37846.1| MFS transporter [Xanthomonas axonopodis pv. citri str. 306] NP_643310.1 0.54 26% ...

  4. NCBI nr-aa BLAST: CBRC-PTRO-23-0017 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PTRO-23-0017 ref|NP_642953.1| phage-related terminase [Xanthomonas axonopodis ...pv. citri str. 306] gb|AAM37489.1| phage-related terminase [Xanthomonas axonopodis pv. citri str. 306] NP_642953.1 5.6 33% ...

  5. NCBI nr-aa BLAST: CBRC-LAFR-01-1513 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-1513 ref|NP_643310.1| MFS transporter [Xanthomonas axonopodis pv. citr...i str. 306] gb|AAM37846.1| MFS transporter [Xanthomonas axonopodis pv. citri str. 306] NP_643310.1 1.1 24% ...

  6. NCBI nr-aa BLAST: CBRC-LAFR-01-1577 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-1577 ref|NP_641723.1| hypothetical protein XAC1388 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM36259.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_641723.1 0.077 39% ...

  7. NCBI nr-aa BLAST: CBRC-DNOV-01-2969 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-2969 ref|NP_641277.1| hypothetical protein XAC0925 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM35813.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_641277.1 3.0 27% ...

  8. NCBI nr-aa BLAST: CBRC-ACAR-01-0078 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-0078 ref|NP_644793.1| avirulence protein [Xanthomonas axonopodis pv. c...itri str. 306] gb|AAM39311.1| avirulence protein [Xanthomonas axonopodis pv. citri str. 306] NP_644793.1 2e-04 26% ...

  9. NCBI nr-aa BLAST: CBRC-XTRO-01-0707 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-0707 ref|NP_644475.1| acetate permease [Xanthomonas axonopodis pv. cit...ri str. 306] gb|AAM39011.1| solute:Na+ symporter [Xanthomonas axonopodis pv. citri str. 306] NP_644475.1 1e-136 79% ...

  10. NCBI nr-aa BLAST: CBRC-DRER-13-0011 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-13-0011 ref|NP_642674.1| hypothetical protein XAC2357 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM37210.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_642674.1 5e-05 30% ...

  11. NCBI nr-aa BLAST: CBRC-XTRO-01-0645 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-0645 ref|NP_642163.1| hypothetical protein XAC1837 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM36699.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_642163.1 9e-42 40% ...

  12. NCBI nr-aa BLAST: CBRC-DYAK-01-0032 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DYAK-01-0032 ref|NP_640378.1| D-3-phosphoglycerate dehydrogenase [Xanthomonas ...axonopodis pv. citri str. 306] gb|AAM34914.1| D-3-phosphoglycerate dehydrogenase [Xanthomonas axonopodis pv. citri str. 306] NP_640378.1 1.1 32% ...

  13. NCBI nr-aa BLAST: CBRC-XTRO-01-0625 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-0625 ref|NP_642734.1| integral membrane protein [Xanthomonas axonopodi...s pv. citri str. 306] gb|AAM37270.1| integral membrane protein [Xanthomonas axonopodis pv. citri str. 306] NP_642734.1 1e-62 27% ...

  14. NCBI nr-aa BLAST: CBRC-ACAR-01-0713 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-0713 ref|NP_641889.1| glucose-galactose transporter [Xanthomonas axono...podis pv. citri str. 306] gb|AAM36425.1| glucose-galactose transporter [Xanthomonas axonopodis pv. citri str. 306] NP_641889.1 4.7 35% ...

  15. NCBI nr-aa BLAST: CBRC-BTAU-01-1628 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-BTAU-01-1628 ref|NP_641751.1| phosphatidate cytidylyltransferase [Xanthomonas ...axonopodis pv. citri str. 306] gb|AAM36287.1| phosphatidate cytidylyltransferase [Xanthomonas axonopodis pv. citri str. 306] NP_641751.1 6.8 24% ...

  16. NCBI nr-aa BLAST: CBRC-DYAK-06-0020 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DYAK-06-0020 ref|NP_642141.1| filamentous haemagglutinin [Xanthomonas axonopod...is pv. citri str. 306] gb|AAM36677.1| filamentous haemagglutinin [Xanthomonas axonopodis pv. citri str. 306] NP_642141.1 0.001 26% ...

  17. NCBI nr-aa BLAST: CBRC-BTAU-01-2358 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-BTAU-01-2358 ref|NP_641879.1| hypothetical protein XAC1546 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM36415.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_641879.1 5.2 31% ...

  18. NCBI nr-aa BLAST: CBRC-ACAR-01-0078 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-0078 ref|NP_644725.1| avirulence protein [Xanthomonas axonopodis pv. c...itri str. 306] gb|AAM39243.1| avirulence protein [Xanthomonas axonopodis pv. citri str. 306] NP_644725.1 2e-04 26% ...

  19. NCBI nr-aa BLAST: CBRC-DRER-13-0009 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-13-0009 ref|NP_642674.1| hypothetical protein XAC2357 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM37210.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_642674.1 5e-05 30% ...

  20. NCBI nr-aa BLAST: CBRC-DDIS-03-0106 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DDIS-03-0106 ref|NP_643514.1| hypothetical protein XAC3206 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM38050.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_643514.1 2e-31 30% ...