Detection of Spiroplasma and Wolbachia in the bacterial gonad community of Chorthippus parallelus.

Science.gov (United States)

Martínez-Rodríguez, P; Hernández-Pérez, M; Bella, J L

2013-07-01

We have recently detected the endosymbiont Wolbachia in multiple individuals and populations of the grasshopper Chorthippus parallelus (Orthoptera: acrididae). This bacterium induces reproductive anomalies, including cytoplasmic incompatibility. Such incompatibilities may help explain the maintenance of two distinct subspecies of this grasshopper, C. parallelus parallelus and C. parallelus erythropus, which are involved in a Pyrenean hybrid zone that has been extensively studied for the past 20 years, becoming a model system for the study of genetic divergence and speciation. To evaluate whether Wolbachia is the sole bacterial infection that might induce reproductive anomalies, the gonadal bacterial community of individuals from 13 distinct populations of C. parallelus was determined by denaturing gradient gel electrophoresis analysis of bacterial 16S rRNA gene fragments and sequencing. The study revealed low bacterial diversity in the gonads: a persistent bacterial trio consistent with Spiroplasma sp. and the two previously described supergroups of Wolbachia (B and F) dominated the gonad microbiota. A further evaluation of the composition of the gonad bacterial communities was carried out by whole cell hybridization. Our results confirm previous studies of the cytological distribution of Wolbachia in C. parallelus gonads and show a homogeneous infection by Spiroplasma. Spiroplasma and Wolbachia cooccurred in some individuals, but there was no significant association of Spiroplasma with a grasshopper's sex or with Wolbachia infection, although subtle trends might be detected with a larger sample size. This information, together with previous experimental crosses of this grasshopper, suggests that Spiroplasma is unlikely to contribute to sex-specific reproductive anomalies; instead, they implicate Wolbachia as the agent of the observed anomalies in C. parallelus.

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

Directory of Open Access Journals (Sweden)

Wei Hu

2018-05-01

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

Honey bee colonies act as reservoirs for two Spiroplasma facultative symbionts and incur complex, multiyear infection dynamics

Science.gov (United States)

Schwarz, Ryan S; Teixeira, Érica Weinstein; Tauber, James P; Birke, Juliane M; Martins, Marta Fonseca; Fonseca, Isabela; Evans, Jay D

2014-01-01

Two species of Spiroplasma (Mollicutes) bacteria were isolated from and described as pathogens of the European honey bee, Apis mellifera, ∼30 years ago but recent information on them is lacking despite global concern to understand bee population declines. Here we provide a comprehensive survey for the prevalence of these two Spiroplasma species in current populations of honey bees using improved molecular diagnostic techniques to assay multiyear colony samples from North America (U.S.A.) and South America (Brazil). Significant annual and seasonal fluctuations of Spiroplasma apis and Spiroplasma melliferum prevalence in colonies from the U.S.A. (n = 616) and Brazil (n = 139) occurred during surveys from 2011 through 2013. Overall, 33% of U.S.A. colonies and 54% of Brazil colonies were infected by Spiroplasma spp., where S. melliferum predominated over S. apis in both countries (25% vs. 14% and 44% vs. 38% frequency, respectively). Colonies were co-infected by both species more frequently than expected in both countries and at a much higher rate in Brazil (52%) compared to the U.S.A. (16.5%). U.S.A. samples showed that both species were prevalent not only during spring, as expected from prior research, but also during other seasons. These findings demonstrate that the model of honey bee spiroplasmas as springtime-restricted pathogens needs to be broadened and their role as occasional pathogens considered in current contexts. PMID:24771723

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

Directory of Open Access Journals (Sweden)

Tian Xia

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

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

Science.gov (United States)

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

2017-12-29

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

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

Science.gov (United States)

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

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

Science.gov (United States)

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

2014-11-01

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

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

Directory of Open Access Journals (Sweden)

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.

Bacterial Infections across the Ants: Frequency and Prevalence of Wolbachia, Spiroplasma, and Asaia

Directory of Open Access Journals (Sweden)

Stefanie Kautz

2013-01-01

Full Text Available Bacterial endosymbionts are common across insects, but we often lack a deeper knowledge of their prevalence across most organisms. Next-generation sequencing approaches can characterize bacterial diversity associated with a host and at the same time facilitate the fast and simultaneous screening of infectious bacteria. In this study, we used 16S rRNA tag encoded amplicon pyrosequencing to survey bacterial communities of 310 samples representing 221 individuals, 176 colonies and 95 species of ants. We found three distinct endosymbiont groups—Wolbachia (Alphaproteobacteria: Rickettsiales, Spiroplasma (Firmicutes: Entomoplasmatales, and relatives of Asaia (Alphaproteobacteria: Rhodospirillales—at different infection frequencies (at the ant species level: 22.1%, 28.4%, and 14.7%, resp. and relative abundances within bacterial communities (1.0%–99.9%. Spiroplasma was particularly enriched in the ant genus Polyrhachis, while Asaia relatives were most prevalent in arboreal ants of the genus Pseudomyrmex. While Wolbachia and Spiroplasma have been surveyed in ants before, Asaia, an acetic acid bacterium capable of fixing atmospheric nitrogen, has received much less attention. Due to sporadic prevalence across all ant taxa investigated, we hypothesize facultative associations for all three bacterial genera. Infection patterns are discussed in relation to potential adaptation of specific bacteria in certain ant groups.

Horizontal Acquisition and Transcriptional Integration of Novel Genes in Mosquito-Associated Spiroplasma.

Science.gov (United States)

Lo, Wen-Sui; Kuo, Chih-Horng

2017-12-01

Genetic differentiation among symbiotic bacteria is important in shaping biodiversity. The genus Spiroplasma contains species occupying diverse niches and is a model system for symbiont evolution. Previous studies have established that two mosquito-associated species have diverged extensively in their carbohydrate metabolism genes despite having a close phylogenetic relationship. Notably, although the commensal Spiroplasma diminutum lacks identifiable pathogenicity factors, the pathogenic Spiroplasma taiwanense was found to have acquired a virulence factor glpO and its associated genes through horizontal transfer. However, it is unclear if these acquired genes have been integrated into the regulatory network. In this study, we inferred the gene content evolution in these bacteria, as well as examined their transcriptomes in response to glucose availability. The results indicated that both species have many more gene acquisitions from the Mycoides-Entomoplasmataceae clade, which contains several important pathogens of ruminants, than previously thought. Moreover, several acquired genes have higher expression levels than the vertically inherited homologs, indicating possible functional replacement. Finally, the virulence factor and its functionally linked genes in S. taiwanense were up-regulated in response to glucose starvation, suggesting that these acquired genes are under expression regulation and the pathogenicity may be a stress response. In summary, although differential gene losses are a major process for symbiont divergence, gene gains are critical in counteracting genome degradation and driving diversification among facultative symbionts. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

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

Science.gov (United States)

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

2015-11-05

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

Complete Genome Sequence of Spiroplasma floricola 23-6T (ATCC 29989), a Bacterium Isolated from a Tulip Tree (Liriodendron tulipifera L.).

Science.gov (United States)

Tsai, Yi-Ming; Wu, Pei-Shan; Lo, Wen-Sui; Kuo, Chih-Horng

2018-04-19

Spiroplasma floricola 23-6 T (ATCC 29989) was isolated from the flower surface of a tulip tree ( Liriodendron tulipifera L.). Here, we report the complete genome sequence of this bacterium to facilitate the investigation of its biology and the comparative genomics among Spiroplasma species. Copyright © 2018 Tsai et al.

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

Science.gov (United States)

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

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

Science.gov (United States)

Chen, Xue Dong; Stelinski, Lukasz L

2017-09-20

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

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.

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.

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

2014-07-01

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

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

Science.gov (United States)

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

2009-01-16

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

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

Science.gov (United States)

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

2017-06-01

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

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.

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.

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

Science.gov (United States)

Langdon, Kevin W; Rogers, Michael E

2017-10-01

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

Innate immune system capabilities of the Asian citrus psyllid, Diaphorina citri.

Science.gov (United States)

Arp, Alex P; Martini, Xavier; Pelz-Stelinski, Kirsten S

2017-09-01

Citrus production worldwide is currently threatened by Huanglongbing, or citrus greening disease. The associated pathogen, Candidatus Liberibacter asiaticus (CLas), is transmitted by the Asian citrus psyllid, Diaphorina citri. Annotation of the D. citri genome revealed a reduced innate immune system lacking a number of antimicrobial peptides and the Imd pathway associated with defense against Gram-negative bacteria. We characterized this apparent immune reduction in survival assays in which D. citri were exposed to Gram-negative or Gram-positive bacteria. D. citri experienced significant mortality when exposed to Serratia marcescens (Gram-negative) through oral ingestion or by septic injury. Escherichia coli (Gram-negative) also caused significant D. citri mortality, but only when inoculated at high concentrations through oral ingestion or by septic injury. Neither Micrococcus luteus (Gram-positive) or Bacillus subtilis (Gram-positive) caused significant mortality as compared to controls in any experiment. E. coli titers increased rapidly following exposure, while M. luteus titer remained stable for 72 h. We demonstrate that D. citri is capable of defending against E. coli, a Gram-negative bacterium, despite lacking the Imd defense pathway. The tolerance of D. citri to M. luteus infection, yet inability to effectively clear infections, presents questions to efficacy of D. citri immune response to effectively clear Gram-positive infections. Copyright © 2017. Published by Elsevier Inc.

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.

The cys regulon of Xanthomonas citri

International Nuclear Information System (INIS)

Moutran, A.; Balan, A.

2012-01-01

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

The cys regulon of Xanthomonas citri

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)

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.

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

Science.gov (United States)

Yan, Qing; Hu, Xiufang; Wang, Nian

2012-10-01

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

Record of Diaphorina citri (Hemiptera: Psyllidae) in North West Argentina

OpenAIRE

Augier, Lucrecia; Gastaminza, Gerardo; Lizondo, Marcelo; Argañaraz, Manuel; Willink, Eduardo

2006-01-01

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.

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

OpenAIRE

Nouri, Shahideh; Salem, Nid?; Falk, Bryce W.

2016-01-01

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.

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

Science.gov (United States)

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

2008-08-01

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

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.

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

Directory of Open Access Journals (Sweden)

Brett R Bayles

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

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

Directory of Open Access Journals (Sweden)

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

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. Copyright © 2016 Nouri et al.

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 citri populations in Iran, India, Saudi Arabia, Brazil, Mexico, Florida and Texas (USA) are similar. Wolbachia, wDi, wsp sequences were similar among Iranian populations, but different between Iranian and Pakistani populations. The South West Asia (SWA) group is the most likely source of the introduced Iranian populations of D. citri. This assertion is also supported by the sequence similarity of the Wolbachia, wDi, strains from the Florida, USA and Iranian D. citri. These results should be considered when looking for biological controls in either country. © 2013 Society of Chemical Industry.

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

Science.gov (United States)

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

2016-05-01

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

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

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

Directory of Open Access Journals (Sweden)

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.

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

Science.gov (United States)

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

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.

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

OpenAIRE

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

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.

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

Science.gov (United States)

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

2017-08-31

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

Disrupting Mating Behavior of Diaphorina citri (Liviidae).

Science.gov (United States)

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

2016-12-01

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

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

Science.gov (United States)

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

2017-02-01

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

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; 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. Copyright © 2016 Matsumura et al.

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

International Nuclear Information System (INIS)

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

2012-01-01

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

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

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)

Genetic variability of Brazilian phytoplasma and spiroplasma isolated from maize plants Variabilidade genética de fitoplasma e espiroplasma isolados de plantas de milho no Brasil

Directory of Open Access Journals (Sweden)

Eliane Aparecida Gomes

2004-01-01

Full Text Available The objective of this work was to characterize the genetic variability of phytoplasma and Spiroplasma kunkelii isolated from maize plants showing symptoms of stunt collected from different Brazilian geographic regions. A DNA fragment of 500 base pairs (bp was amplified from the spiralin gene in S. kunkelii and one fragment of 1,200 bp was generated from 16S rDNA gene in phytoplasma. The partial sequences of the spiralin gene showed similarity of 98% among the isolates of S. kunkelii analyzed. These sequences were compared with the sequence of the spiralin gene from other Spiroplasma species deposited in the GenBank, resulting in a similarity varying from 76.9% to 88.1%. The 16S rDNA sequence from the phytoplasma were completely similar within the Brazilian isolates and showed up to 98% of the similarity with sequences already found from other phytoplasmas. A very narrow genetic variability was detected by these gene fragments within phytoplasma and Spiroplasma analyzed. However, other genomic regions with higher polymorphic levels shall be identified in order to better evaluate the genetic diversity within these microorganisms population.O objetivo deste trabalho foi caracterizar a variabilidade genética de isolados de fitoplasma e de Spiroplasma kunkelii obtidos de plantas de milho, apresentando sintomas de enfezamento, coletados em diferentes regiões do Brasil. Um fragmento de 500 pares de bases (pb do gene que codifica a espiralina de S. kunkelii foi amplificado e um produto de amplificação de 1.200 pb foi gerado a partir do gene 16S rDNA de fitoplasma. As seqüências parciais do gene da espiralina mostraram similaridade de 98% entre os isolados de S. kunkelii analisados. Essas seqüências foram comparadas com a seqüência do gene da espiralina de outras espécies de Spiroplasma depositadas no GenBank, resultando em similaridade variável entre 76,9% e 88,1%. As seqüências do gene 16S rDNA dos isolados de fitoplasma foram

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

Science.gov (United States)

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

2018-05-03

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

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

Directory of Open Access Journals (Sweden)

I NYOMAN WIJAYA

2015-09-01

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

Analysis of transcriptome differences between resistant and susceptible strains of the citrus red mite Panonychus citri (Acari: Tetranychidae.

Directory of Open Access Journals (Sweden)

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

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

Science.gov (United States)

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

2017-10-01

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

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

Science.gov (United States)

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

2015-02-01

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

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

Science.gov (United States)

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

2011-02-01

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

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

Science.gov (United States)

Tabachnick, Walter J

2015-06-01

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

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

Science.gov (United States)

Vankosky, Meghan A; Hoddle, Mark S

2017-06-07

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

Influence of gamma radiation on antibodies fixation on polystyrene. Application to ELISA enzyme immunoassay

International Nuclear Information System (INIS)

Esterlin, S.

1986-01-01

This thesis is divided into two parts: the first part includes a description of the ELISA test, a comparative analysis of the main supports (polystyrene microtitration trays) used for this technique and an evaluation of gamma radiation effects on the quality of the supports. The study was carried out with the following antigens: Spiroplasma citri R8A2, a tymovirus and Tristeza virus. The second part dealt with the effects of gamma radiations on antibodies and antibody-support system (interaction immobilized immunoglobulins-plastic support) [fr

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

Science.gov (United States)

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

2013-02-01

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

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

Science.gov (United States)

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

2015-01-01

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

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

Science.gov (United States)

Sétamou, Mamoudou; Bartels, David W

2015-01-01

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

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

Directory of Open Access Journals (Sweden)

Mamoudou Sétamou

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

Effects of Mineral Oils on Host Selection Behavior of Diaphorina citri

Directory of Open Access Journals (Sweden)

Mofit Eko Poerwanto

2008-07-01

Full Text Available Response of Diaphorina citri Kuwayama (Hemiptera: Psyllidae on citrus leaves treated with horticultural and agricultural mineral oil (HMO & AMO was evaluated under laboratory condition in choice and non-choice trials. Combination of HMO and AMO at concentration of 0 up to 2% was applied to citrus leaves. Mineral oil applications significantly reduced the proportion of D. citri stayed on citrus leaves for feeding with the proposed mechanisms: 1 oils covered leaves thus hampered it for releasing volatile compounds, 2 oil film preventing the direct contact of tarsus or palpus to leaf surfaces, and 3 the volatile compounds produced by mineral oils also blended with plant volatiles in the atmosphere and reduced the probability of olfactory receptors capturing the olfactory cues for locating the feeding site. Mineral oils also increased the mortality of D. citri due to the refusion for feeding on the treated citrus leaves. The effect of AMO was more prominent than HMO.   Tanggapan Diaphorina citri Kuwayama (Hemiptera: Psyllidae pada daun jeruk yang diperlakukan dengan minyak mineral (HMO dan AMO dievaluasi dengan metode pilihan maupun tanpa pilihan di laboratorium. Kombinasi antara HMO dan AMO pada konsentrasi 0% sampai dengan 2% diaplikasikan pada daun jeruk. Minyak mineral secara nyata menurunkan proporsi D. citri hinggap untuk makan pada daun jeruk, yang mungkin disebabkan oleh mekanisme: 1 minyak melapisi daun sehingga tidak mampu melepaskan senyawa volatil, 2 lapisan minyak menghalangi kontak langsung antara tarsus atau palpus pada permukaan daun, dan 3 senyawa volatil dari minyak bercampur dengan volatil tanaman di udara sehingga menurunkan probabilitas reseptor olfaktori menangkap sinyal penanda lokasi inang. Minyak mineral juga meningkatkan mortalitas, akibat dari penolakan makan pada daun jeruk yang diperlakukan. Pengaruh AMO lebih besar dari pada HMO.

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

Energy Technology Data Exchange (ETDEWEB)

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

2011-03-22

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

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

Directory of Open Access Journals (Sweden)

Shijiang Yu

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

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

Science.gov (United States)

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

2016-01-01

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

Processed kaolin affects the probing and settling behavior of Diaphorina citri (Hemiptera: Lividae).

Science.gov (United States)

Miranda, Marcelo P; Zanardi, Odimar Z; Tomaseto, Arthur F; Volpe, Haroldo Xl; Garcia, Rafael B; Prado, Ernesto

2018-03-05

Alternative methods that have the potential to reduce the entry of Diaphorina citri Kuwayama (Hemiptera: Liviidae), the major citrus pest worldwide, into commercial groves could be a viable approach for huanglongbing management. Kaolin is an aluminum silicate that when sprayed on plants forms a white particle film that interferes with host recognition by the insects. Diaphorina citri orients towards the host plants by visual and olfactory cues. The purpose of the study was to determine the effect of processed kaolin on D. citri settling (no-choice) and probing behavior [electrical penetration graph (EPG) technique] under laboratory conditions, and to study its host plant finding ability and dispersal under field conditions in the absence and presence of young shoots. Under laboratory conditions, kaolin caused an overall reduction of 40% in the number of psyllids settled on treated seedlings; furthermore, the proportion of individuals that were able to reach the phloem was 50% lower on kaolin-treated plants than on untreated plants. In the field, the plant finding ability of D. citri was disrupted on kaolin-treated trees (overall reduction of 96%), regardless of the vegetative condition, and psyllid dispersal was slower in kaolin-treated plots than in the untreated control. This study clearly demonstrates that processed kaolin interferes negatively with different aspects of the host plant finding ability of D. citri. These findings suggest that processed kaolin has a high potential to reduce huanglongbing primary infection. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

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

Science.gov (United States)

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

2017-02-01

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

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

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.

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

OpenAIRE

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

2016-01-01

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.

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

Science.gov (United States)

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

2011-01-01

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

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. Copyright © 2016 Nigg et al.

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

Science.gov (United States)

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

2012-10-01

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

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

Science.gov (United States)

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

2018-02-01

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

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

Science.gov (United States)

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

2013-10-01

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

The monomeric, tetrameric, and fibrillar organization of Fib: the dynamic building block of the bacterial linear motor of Spiroplasma melliferum BC3.

Science.gov (United States)

Cohen-Krausz, Sara; Cabahug, Pamela C; Trachtenberg, Shlomo

2011-07-08

Spiroplasmas belong to the class Mollicutes, representing the minimal, free-living, and self-replicating forms of life. Spiroplasmas are helical wall-less bacteria and the only ones known to swim by means of a linear motor (rather than the near-universal rotary bacterial motor). The linear motor follows the shortest path along the cell's helical membranal tube. The motor is composed of a flat monolayered ribbon of seven parallel fibrils and is believed to function in controlling cell helicity and motility through dynamic, coordinated, differential length changes in the fibrils. The latter cause local perturbations of helical symmetry, which are essential for net directional displacement in environments with a low Reynolds number. The underlying fibrils' core building block is a circular tetramer of the 59-kDa protein Fib. The fibrils' differential length changes are believed to be driven by molecular switching of Fib, leading consequently to axial ratio and length changes in tetrameric rings. Using cryo electron microscopy, diffractometry, single-particle analysis of isolated ribbons, and sequence analyses of Fib, we determined the overall molecular organization of the Fib monomer, tetramer, fibril, and linear motor of Spiroplasma melliferum BC3 that underlies cell geometry and motility. Fib appears to be a bidomained molecule, of which the N-terminal half is apparently a globular phosphorylase. By a combination of reversible rotation and diagonal shift of Fib monomers, the tetramer adopts either a cross-like nonhanded conformation or a ring-like handed conformation. The sense of Fib rotation may determine the handedness of the linear motor and, eventually, of the cell. A further change in the axial ratio of the ring-like tetramers controls fibril lengths and the consequent helical geometry. Analysis of tetramer quadrants from adjacent fibrils clearly demonstrates local differential fibril lengths. Copyright © 2011 Elsevier Ltd. All rights reserved.

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 ...

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.

Citrus flush shoot ontogeny modulates biotic potential of Diaphorina citri.

Science.gov (United States)

Cifuentes-Arenas, Juan Camilo; de Goes, António; de Miranda, Marcelo Pedreira; Beattie, George Andrew Charles; Lopes, Silvio Aparecido

2018-01-01

The biology and behaviour of the psyllid Diaphorina citri Kuwayama (Hemiptera: Sternorrhyncha: Liviidae), the major insect vector of bacteria associated with huanglongbing, have been extensively studied with respect to host preferences, thermal requirements, and responses to visual and chemical volatile stimuli. However, development of the psyllid in relation to the ontogeny of immature citrus flush growth has not been clearly defined or illustrated. Such information is important for determining the timing and frequency of measures used to minimize populations of the psyllid in orchards and spread of HLB. Our objective was to study how flush ontogeny influences the biotic potential of the psyllid. We divided citrus flush growth into six stages within four developmental phases: emergence (V1), development (V2 and V3), maturation (V4 and V5), and dormancy (V6). Diaphorina citri oviposition and nymph development were assessed on all flush stages in a temperature controlled room, and in a screen-house in which ambient temperatures varied. Our results show that biotic potential of Diaphorina citri is not a matter of the size or the age of the flushes (days after budbreak), but the developmental stage within its ontogeny. Females laid eggs on flush V1 to V5 only, with the time needed to commence oviposition increasing with the increasing in flush age. Stages V1, V2 and V3 were most suitable for oviposition, nymph survival and development, and adult emergence, which showed evidence of protandry. Flush shoots at emerging and developmental phases should be the focus of any chemical or biological control strategy to reduce the biotic potential of D. citri, to protect citrus tree from Liberibacter infection and to minimize HLB dissemination.

Gross morphology and ultrastructure of the female reproductive system of Diaphorina citri (Hemiptera: Liviidae

Directory of Open Access Journals (Sweden)

Fabio Cleisto Alda Dossi

2014-04-01

Full Text Available The morphological traits of the female reproductive system of Diaphorina citri were examined in detail. Diaphorina citri has telotrophic ovaries with ovarioles organized as a "bouquet", displaying a rudimentary terminal filament and a syncytial tropharium. The vitellarium carries a single growing oocyte at each maturation cycle, which is connected with the tropharium by a nutritive cord. Morpho-functional changes occur during oocyte development, mainly during mid to late vitellogenesis. Morphological events such as the patency of the follicular cells and the intense traffic of vesicles through para- or intracellular processes, suggest a possible route for endosymbiont invasion of D. citri reproductive tissues. Similar events have been demonstrated to be involved in the process of ovariole invasion by endosymbionts in other sternorrhynchans that share reproductive traits with psyllids.

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

Science.gov (United States)

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

2017-11-01

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

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...

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

Science.gov (United States)

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

2015-12-16

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

Mycoplasmas, plants, insect vectors: a matrimonial triangle.

Science.gov (United States)

Garnier, M; Foissac, X; Gaurivaud, P; Laigret, F; Renaudin, J; Saillard, C; Bové, J M

2001-10-01

Plant pathogenic mycoplasmas were discovered by electron microscopy, in 1967, long after the discovery and culture in 1898 of the first pathogenic mycoplasma of animal origin, Mycoplasma mycoides. Mycoplasmas are Eubacteria of the class Mollicutes, a group of organisms phylogenetically related to Gram-positive bacteria. Their more characteristic features reside in the small size of their genomes, the low guanine (G) plus cytosine (C) content of their genomic DNA and the lack of a cell wall. Plant pathogenic mycoplasmas are responsible for several hundred diseases and belong to two groups: the phytoplasmas and the spiroplasmas. The phytoplasmas (previously called MLOs, for mycoplasma like organisms) were discovered first; they are pleiomorphic, and have so far resisted in vitro cultivation. Phytoplasmas represent the largest group of plant pathogenic Mollicutes. Only three plant pathogenic spiroplasmas are known today. Spiroplasma citri, the agent of citrus stubborn was discovered and cultured in 1970 and shown to be helical and motile. S. kunkelii is the causal agent of corn stunt. S. phoeniceum, responsible for periwinkle yellows, was discovered in Syria. There are many other spiroplasmas associated with insects and ticks. Plant pathogenic mycoplasmas are restricted to the phloem sieve tubes in which circulates the photosynthetically-enriched sap, the food for many phloem-feeding insects (aphids, leafhoppers, psyllids, etc.). Interestingly, phytopathogenic mycoplasmas are very specifically transmitted by leafhoppers or psyllid species. In this paper, the most recent knowledge on phytopathogenic mycoplasmas in relation with their insect and plant habitats is presented as well as the experiments carried out to control plant mycoplasma diseases, by expression of mycoplasma-directed-antibodies in plants (plantibodies).

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

Science.gov (United States)

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

2018-01-24

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

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

Science.gov (United States)

Killiny, Nabil; Nehela, Yasser

2017-07-01

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

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

Directory of Open Access Journals (Sweden)

José Bernardo Navarrete Cedeño

2016-12-01

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

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

Directory of Open Access Journals (Sweden)

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.

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

Science.gov (United States)

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

2017-05-01

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

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

Science.gov (United States)

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

2014-01-01

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

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

Directory of Open Access Journals (Sweden)

Nabil Killiny

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

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

Science.gov (United States)

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

2014-01-01

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

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

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.

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

Science.gov (United States)

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

2016-09-01

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

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.

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

Science.gov (United States)

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

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

Science.gov (United States)

Pelz-Stelinski, K S; Killiny, N

2016-05-01

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

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

Science.gov (United States)

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

2018-04-01

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

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

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

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

Science.gov (United States)

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

2017-12-05

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

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

African Journals Online (AJOL)

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

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

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

Science.gov (United States)

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

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

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

Male Diaphorina citri searching responses to vibrational communication signals

Science.gov (United States)

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

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

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

Science.gov (United States)

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

2017-02-01

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

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

OpenAIRE

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

2011-01-01

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

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

Directory of Open Access Journals (Sweden)

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.

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.

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

Science.gov (United States)

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

2016-01-01

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

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.

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.

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

Science.gov (United States)

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

2015-06-01

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

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

Science.gov (United States)

Dossi, Fábio C A; Cônsoli, Fernando L

2010-01-01

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

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.

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 ...

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

Science.gov (United States)

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

2018-03-25

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

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

Science.gov (United States)

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

2017-02-01

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

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.

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

Directory of Open Access Journals (Sweden)

Ebratt Ravelo Everth Emilio

2011-12-01

Full Text Available

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

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

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

Science.gov (United States)

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

2015-12-01

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

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

Science.gov (United States)

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

2018-03-25

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

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

Science.gov (United States)

Yu, Xiudao; Gowda, Siddarame; Killiny, Nabil

2017-09-01

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

Pengendalian Diaphorina citri (Vektor Penyakit CVPD dengan Metarrhizium anisopliae

Directory of Open Access Journals (Sweden)

Kardi Raharjo

2000-07-01

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

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

Science.gov (United States)

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

2017-03-01

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

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

Science.gov (United States)

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

2015-06-01

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

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.

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

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

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

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

Science.gov (United States)

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

2015-02-01

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

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

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.

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

Directory of Open Access Journals (Sweden)

Mizu lstianto

2003-12-01

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

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

International Nuclear Information System (INIS)

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

2012-01-01

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

A structural framework for a near-minimal form of life: mass and compositional analysis of the helical mollicute Spiroplasma melliferum BC3.

Directory of Open Access Journals (Sweden)

Shlomo Trachtenberg

Full Text Available Spiroplasma melliferum is a wall-less bacterium with dynamic helical geometry. This organism is geometrically well defined and internally well ordered, and has an exceedingly small genome. Individual cells are chemotactic, polar, and swim actively. Their dynamic helicity can be traced at the molecular level to a highly ordered linear motor (composed essentially of the proteins fib and MreB that is positioned on a defined helical line along the internal face of the cell's membrane. Using an array of complementary, informationally overlapping approaches, we have taken advantage of this uniquely simple, near-minimal life-form and its helical geometry to analyze the copy numbers of Spiroplasma's essential parts, as well as to elucidate how these components are spatially organized to subserve the whole living cell. Scanning transmission electron microscopy (STEM was used to measure the mass-per-length and mass-per-area of whole cells, membrane fractions, intact cytoskeletons and cytoskeletal components. These local data were fit into whole-cell geometric parameters determined by a variety of light microscopy modalities. Hydrodynamic data obtained by analytical ultracentrifugation allowed computation of the hydration state of whole living cells, for which the relative amounts of protein, lipid, carbohydrate, DNA, and RNA were also estimated analytically. Finally, ribosome and RNA content, genome size and gene expression were also estimated (using stereology, spectroscopy and 2D-gel analysis, respectively. Taken together, the results provide a general framework for a minimal inventory and arrangement of the major cellular components needed to support life.

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.

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

Directory of Open Access Journals (Sweden)

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.

Repellency of Selected Psidium guajava cultivars to the Asian citrus psyllid, Diaphorina citri

Science.gov (United States)

Asiatic huanglongbing (HLB)(also known as citrus greening disease) is the most devastating disease of citrus worldwide. It is caused by a bacterium ‘Candidatus Liberibacter asiaticus’ and transmitted by the Asian citrus psyllid (ACP), Diaphorina citri. Considerable research has been conducted toward...

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

2017-04-01

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

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.

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

Science.gov (United States)

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

2017-09-01

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

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

Science.gov (United States)

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

2017-03-01

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

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

Directory of Open Access Journals (Sweden)

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.

Responses of Diaphorina citri (Hemiptera: Psyllidae) to conspecific vibrational signals and synthetic mimics

Science.gov (United States)

Mate-seeking in Diaphorina citri Kuwayama, a vector of the economically damaging huanglongbing citrus disease, typically includes male-female duetting behaviors. First, the male calls by beating its wings at ca. 170-250 Hz, producing vibrations that are transmitted along the host tree branches to th...

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

International Nuclear Information System (INIS)

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

1982-01-01

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

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

Directory of Open Access Journals (Sweden)

Wijaya I N.

2012-10-01

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

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

Directory of Open Access Journals (Sweden)

Vivek Kumar

2017-10-01

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

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

Science.gov (United States)

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

2017-10-31

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

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

Science.gov (United States)

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

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

Science.gov (United States)

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

2018-01-01

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

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

International Nuclear Information System (INIS)

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

2009-01-01

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

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

Science.gov (United States)

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

2009-12-01

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

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.

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

Directory of Open Access Journals (Sweden)

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.

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

Science.gov (United States)

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

2018-04-02

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

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

Science.gov (United States)

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

2018-01-01

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

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

Science.gov (United States)

Yu, Xiudao; Killiny, Nabil

2018-03-01

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

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

Science.gov (United States)

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

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

NARCIS (Netherlands)

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

2017-01-01

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

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

Science.gov (United States)

2008-01-01

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

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

Science.gov (United States)

Hoddle, Mark S; Pandey, Raju

2014-02-01

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

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

Science.gov (United States)

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

2018-04-01

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

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

Science.gov (United States)

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

2017-08-01

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

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

Science.gov (United States)

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

2016-10-01

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

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

Science.gov (United States)

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

2006-05-01

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

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

NARCIS (Netherlands)

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

2017-01-01

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

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

International Nuclear Information System (INIS)

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

2006-01-01

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2006-03-01

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

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

Science.gov (United States)

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

2016-12-01

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

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

Directory of Open Access Journals (Sweden)

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.

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

Science.gov (United States)

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

2018-01-01

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

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

Science.gov (United States)

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

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)

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

2015-01-01

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

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

Science.gov (United States)

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

Genomic survey of pathogenicity determinants and VNTR markers in the cassava bacterial pathogen Xanthomonas axonopodis pv. Manihotis strain CIO151.

Science.gov (United States)

Arrieta-Ortiz, Mario L; Rodríguez-R, Luis M; Pérez-Quintero, Álvaro L; Poulin, Lucie; Díaz, Ana C; Arias Rojas, Nathalia; Trujillo, Cesar; Restrepo Benavides, Mariana; Bart, Rebecca; Boch, Jens; Boureau, Tristan; Darrasse, Armelle; David, Perrine; Dugé de Bernonville, Thomas; Fontanilla, Paula; Gagnevin, Lionel; Guérin, Fabien; Jacques, Marie-Agnès; Lauber, Emmanuelle; Lefeuvre, Pierre; Medina, Cesar; Medina, Edgar; Montenegro, Nathaly; Muñoz Bodnar, Alejandra; Noël, Laurent D; Ortiz Quiñones, Juan F; Osorio, Daniela; Pardo, Carolina; Patil, Prabhu B; Poussier, Stéphane; Pruvost, Olivier; Robène-Soustrade, Isabelle; Ryan, Robert P; Tabima, Javier; Urrego Morales, Oscar G; Vernière, Christian; Carrere, Sébastien; Verdier, Valérie; Szurek, Boris; Restrepo, Silvia; López, Camilo; Koebnik, Ralf; Bernal, Adriana

2013-01-01

Xanthomonas axonopodis pv. manihotis (Xam) is the causal agent of bacterial blight of cassava, which is among the main components of human diet in Africa and South America. Current information about the molecular pathogenicity factors involved in the infection process of this organism is limited. Previous studies in other bacteria in this genus suggest that advanced draft genome sequences are valuable resources for molecular studies on their interaction with plants and could provide valuable tools for diagnostics and detection. Here we have generated the first manually annotated high-quality draft genome sequence of Xam strain CIO151. Its genomic structure is similar to that of other xanthomonads, especially Xanthomonas euvesicatoria and Xanthomonas citri pv. citri species. Several putative pathogenicity factors were identified, including type III effectors, cell wall-degrading enzymes and clusters encoding protein secretion systems. Specific characteristics in this genome include changes in the xanthomonadin cluster that could explain the lack of typical yellow color in all strains of this pathovar and the presence of 50 regions in the genome with atypical nucleotide composition. The genome sequence was used to predict and evaluate 22 variable number of tandem repeat (VNTR) loci that were subsequently demonstrated as polymorphic in representative Xam strains. Our results demonstrate that Xanthomonas axonopodis pv. manihotis strain CIO151 possesses ten clusters of pathogenicity factors conserved within the genus Xanthomonas. We report 126 genes that are potentially unique to Xam, as well as potential horizontal transfer events in the history of the genome. The relation of these regions with virulence and pathogenicity could explain several aspects of the biology of this pathogen, including its ability to colonize both vascular and non-vascular tissues of cassava plants. A set of 16 robust, polymorphic VNTR loci will be useful to develop a multi-locus VNTR analysis

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...

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.

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

Science.gov (United States)

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

2014-11-01

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

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

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.

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.

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...

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

Science.gov (United States)

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

2018-06-01

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

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

Science.gov (United States)

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

2015-01-01

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

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

Science.gov (United States)

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

2017-12-05

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

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

Science.gov (United States)

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

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

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...

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

Directory of Open Access Journals (Sweden)

D. Marian Szebenyi

2018-04-01

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

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

Science.gov (United States)

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

2015-06-01

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

Isolation of radiation resistant fungal strains from highly radioactive field

International Nuclear Information System (INIS)

Adam, Y.M.; Aziz, N.H.; Attaby, H.S.H.

1995-01-01

This study examined the radiation resistance of fungal flora isolated from the hot-lab around the radiation sources, cobalt 137 and radium 226 . The predominant mould species were: Aspergillus flavus, A. Niger, penicillium chrysogenum, cladosporium herbarum, fusarium oxysporum and alternaria citri. The D 10 values of F. Oxysporum; 2.00 KGy, A. Flavus; 1.40 KGy, P. chrysogenum; 1.15 KGy, and A. citri; 0.95 KGy, are about 1.67, 3.10, 1.92 and 1.36 folds as the D 1 0 values of the same isolates recovered from soil

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

Science.gov (United States)

Luo, Yufa; Agnarsson, Ingi

2018-01-01

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

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

Science.gov (United States)

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

2018-04-01

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2012-07-01

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

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

International Nuclear Information System (INIS)

Tambascia, C.; Balan, A.

2012-01-01

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

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

Science.gov (United States)

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

2014-04-01

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

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

Science.gov (United States)

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

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...

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...

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

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

Science.gov (United States)

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

2012-01-01

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

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...

Gut microbiota of Tenebrio molitor and their response to environmental change.

Science.gov (United States)

Jung, Jaejoon; Heo, Aram; Park, Yong Woo; Kim, Ye Ji; Koh, Hyelim; Park, Woojun

2014-07-01

A bacterial community analysis of the gut of Tenebrio molitor larvae was performed using pyrosequencing of the 16S rRNA gene. A predominance of genus Spiroplasma species in phylum Tenericutes was observed in the gut samples, but there was variation found in the community composition between T. molitor individuals. The gut bacteria community structure was not significantly affected by the presence of antibiotics or by the exposure of T. molitor larvae to a highly diverse soil bacteria community. A negative relationship was identified between bacterial diversity and ampicillin concentration; however, no negative relationship was identified with the addition of kanamycin. Ampicillin treatment resulted in a reduction in the bacterial community size, estimated using the 16S rRNA gene copy number. A detailed phylogenetic analysis indicated that the Spiroplasma-associated sequences originating from the T. molitor larvae were distinct from previously identified Spiroplasma type species, implying the presence of novel Spiroplasma species. Some Spiroplasma species are known to be insect pathogens; however, the T. molitor larvae did not experience any harmful effects arising from the presence of Spiroplasma species, indicating that Spiroplasma in the gut of T. molitor larvae do not act as a pathogen to the host. A comparison with the bacterial communities found in other insects (Apis and Solenopsis) showed that the Spiroplasma species found in this study were specific to T. molitor.

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.

Multilocus Sequence Typing Reveals Relevant Genetic Variation and Different Evolutionary Dynamics among Strains of Xanthomonas arboricola pv. juglandis

Directory of Open Access Journals (Sweden)

Marco Scortichini

2010-11-01

Full Text Available Forty-five Xanthomonas arboricola pv. juglandis (Xaj strains originating from Juglans regia cultivation in different countries were molecularly typed by means of MultiLocus Sequence Typing (MLST, using acnB, gapA, gyrB and rpoD gene fragments. A total of 2.5 kilobases was used to infer the phylogenetic relationship among the strains and possible recombination events. Haplotype diversity, linkage disequilibrium analysis, selection tests, gene flow estimates and codon adaptation index were also assessed. The dendrograms built by maximum likelihood with concatenated nucleotide and amino acid sequences revealed two major and two minor phylotypes. The same haplotype was found in strains originating from different continents, and different haplotypes were found in strains isolated in the same year from the same location. A recombination breakpoint was detected within the rpoD gene fragment. At the pathovar level, the Xaj populations studied here are clonal and under neutral selection. However, four Xaj strains isolated from walnut fruits with apical necrosis are under diversifying selection, suggesting a possible new adaptation. Gene flow estimates do not support the hypothesis of geographic isolation of the strains, even though the genetic diversity between the strains increases as the geographic distance between them increases. A triplet deletion, causing the absence of valine, was found in the rpoD fragment of all 45 Xaj strains when compared with X. axonopodis pv. citri strain 306. The codon adaptation index was high in all four genes studied, indicating a relevant metabolic activity.

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

Science.gov (United States)

Killiny, Nabil; Kishk, Abdelaziz

2017-06-01

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

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2006-05-01

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

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

Directory of Open Access Journals (Sweden)

Bei Gao

2011-05-01

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

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.

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

Science.gov (United States)

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

2012-07-01

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

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

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

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...

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

Directory of Open Access Journals (Sweden)

Joseph M Patt

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

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

Science.gov (United States)

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

2018-01-01

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

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

Science.gov (United States)

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

2018-01-01

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

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

Science.gov (United States)

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

2015-10-01

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

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

Directory of Open Access Journals (Sweden)

Adriano Farias

2018-04-01

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

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.

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.

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

Science.gov (United States)

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

2016-01-01

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

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

Directory of Open Access Journals (Sweden)

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

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

Directory of Open Access Journals (Sweden)

Lenira Viana Costa Santa-Cecília

2013-03-01

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

Isolation and identification of citrus psorosis virus Egyptian isolate (CPsV-EG).

Science.gov (United States)

Ghazal, S A; El-Dougdoug, Kh A; Mousa, A A; Fahmy, H; Sofy, A R

2008-01-01

Citrus psorosis ophiovirus (CPsV), is considered to be of the most serious and deter mental virus pathogen's citrus species trees in Egypt. CPsV-EG was isolated from infected citrus grapefruit (C. paradisi Macf.) at Agric. Res. Centre (ARC). The grapefruit which used for CPsV-EG isolate was found to be free from CTV, CEVd and Spiroplasma citri where as gave -ve results with DTBIA, tissue print hybridization and Diene's stain respectively. CPsV-EG was detected on the basis of biological indexing by graft inoculation which gave oak leaf pattern (OLP) on Dweet tangor and serological assay by DAS-ELISA using Mab specific CPsV. CPsV-EG was reacted with variable responses on 16 host plants belonging to 6 families. Only 8 host plants are susceptible and showed visible external symptoms which appeared as local, systemic and local followed by systemic infections. CPsV-EG isolate was transmitted from infected citrus to citrus by syringe and grafting and herbaceous plants by forefinger inoculation and syringe. The woody indicators and rootstocks were differed in response to CPsV-EG isolate which appeared as no-response, response, sensitivity and hypersensitivity. The serological characters represented as the antigenic determinants of CPsV-EG isolate related to monoclonal antibodies specific CPsV strain where as appeared precipitation reaction by DAS-ELISA and DTBIA. The partial fragment of RNA3 (coat protein gene) of CPsV-EG (-1140bp and -571bp) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from grapefruit tissues using two sets primers specific CPsV (CPV3 and CPV4) and (PS66 and PS65) respectively. The virus under study was identified as CPsV-EG isolate according to biological, serological and molecular characters.

Prevalence of pathogenic bacteria in Ixodes ricinus ticks in Central Bohemia.

Science.gov (United States)

Klubal, Radek; Kopecky, Jan; Nesvorna, Marta; Sparagano, Olivier A E; Thomayerova, Jana; Hubert, Jan

2016-01-01

Bacteria associated with the tick Ixodes ricinus were assessed in specimens unattached or attached to the skin of cats, dogs and humans, collected in the Czech Republic. The bacteria were detected by PCR in 97 of 142 pooled samples including 204 ticks, i.e. 1-7 ticks per sample, collected at the same time from one host. A fragment of the bacterial 16S rRNA gene was amplified, cloned and sequenced from 32 randomly selected samples. The most frequent sequences were those related to Candidatus Midichloria midichlori (71% of cloned sequences), followed by Diplorickettsia (13%), Spiroplasma (3%), Rickettsia (3%), Pasteurella (3%), Morganella (3%), Pseudomonas (2%), Bacillus (1%), Methylobacterium (1%) and Phyllobacterium (1%). The phylogenetic analysis of Spiroplasma 16S rRNA gene sequences showed two groups related to Spiroplasma eriocheiris and Spiroplasma melliferum, respectively. Using group-specific primers, the following potentially pathogenic bacteria were detected: Borellia (in 20% of the 142 samples), Rickettsia (12%), Spiroplasma (5%), Diplorickettsia (5%) and Anaplasma (2%). In total, 68% of I. ricinus samples (97/142) contained detectable bacteria and 13% contained two or more putative pathogenic groups. The prevalence of tick-borne bacteria was similar to the observations in other European countries.

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...

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

A Nightmare for Males? A Maternally Transmitted Male-Killing Bacterium and Strong Female Bias in a Green Lacewing Population.

Directory of Open Access Journals (Sweden)

Masayuki Hayashi

Full Text Available For maternally transmitted microbes, a female-biased host sex ratio is of reproductive advantage. Here we found a strong female bias in a field population of the green lacewing, Mallada desjardinsi (Insecta; Neuroptera. This bias was attributed to the predominance of individuals harboring a maternally inherited male-killing bacterium that was phylogenetically closely related to the plant-pathogenic Spiroplasma phoeniceum and Spiroplasma kunkelii. Among 35 laboratory-reared broods produced by wild-caught females, 21 broods (60%-all infected with Spiroplasma-consisted of only females (940 individuals. Among 14 broods consisting of both males and females (516 and 635 individuals, respectively, 4 broods were doubly infected with Spiroplasma and Rickettsia, 6 broods were singly infected with Rickettsia, and 3 broods were uninfected (remaining one brood was unknown. Mortality during embryonic and larval development was prominent in all-female broods but not in normal sex ratio broods. Following antibiotic treatment on all-female broods, mortality was significantly reduced and the sex ratio was restored to 1:1. Strong expression and high prevalence of this male-killer is remarkable considering its low density (~10-5-10-4 cells per host mitochondrial gene copy based on quantitative PCR. In addition, a bacterium closely related to Rickettsia bellii was present in 25 of 34 broods (73.5%, irrespective of the sex ratio, with the infection density comparable to other cases of endosymbiosis (~10-2-10-1 cells per mitochondrial gene copy. Higher density of Rickettsia than Spiroplasma was also demonstrated by electron microscopy which visualized both Spiroplasma-like cells and Rickettsia-like cells inside and outside the ovarian cells.

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

Directory of Open Access Journals (Sweden)

Iliano V Coutinho-Abreu

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

Endosymbiont-based immunity in Drosophila melanogaster against parasitic nematode infection.

Science.gov (United States)

Yadav, Shruti; Frazer, Joanna; Banga, Ashima; Pruitt, Katherine; Harsh, Sneh; Jaenike, John; Eleftherianos, Ioannis

2018-01-01

Associations between endosymbiotic bacteria and their hosts represent a complex ecosystem within organisms ranging from humans to protozoa. Drosophila species are known to naturally harbor Wolbachia and Spiroplasma endosymbionts, which play a protective role against certain microbial infections. Here, we investigated whether the presence or absence of endosymbionts affects the immune response of Drosophila melanogaster larvae to infection by Steinernema carpocapsae nematodes carrying or lacking their mutualistic Gram-negative bacteria Xenorhabdus nematophila (symbiotic or axenic nematodes, respectively). We find that the presence of Wolbachia alone or together with Spiroplasma promotes the survival of larvae in response to infection with S. carpocapsae symbiotic nematodes, but not against axenic nematodes. We also find that Wolbachia numbers are reduced in Spiroplasma-free larvae infected with axenic compared to symbiotic nematodes, and they are also reduced in Spiroplasma-containing compared to Spiroplasma-free larvae infected with axenic nematodes. We further show that S. carpocapsae axenic nematode infection induces the Toll pathway in the absence of Wolbachia, and that symbiotic nematode infection leads to increased phenoloxidase activity in D. melanogaster larvae devoid of endosymbionts. Finally, infection with either type of nematode alters the metabolic status and the fat body lipid droplet size in D. melanogaster larvae containing only Wolbachia or both endosymbionts. Our results suggest an interaction between Wolbachia endosymbionts with the immune response of D. melanogaster against infection with the entomopathogenic nematodes S. carpocapsae. Results from this study indicate a complex interplay between insect hosts, endosymbiotic microbes and pathogenic organisms.

Small RNA profiling reveals phosphorus deficiency as a contributing factor in symptom expression for citrus huanglongbing disease.

Science.gov (United States)

Zhao, Hongwei; Sun, Ruobai; Albrecht, Ute; Padmanabhan, Chellappan; Wang, Airong; Coffey, Michael D; Girke, Thomas; Wang, Zonghua; Close, Timothy J; Roose, Mikeal; Yokomi, Raymond K; Folimonova, Svetlana; Vidalakis, Georgios; Rouse, Robert; Bowman, Kim D; Jin, Hailing

2013-03-01

Huanglongbing (HLB) is a devastating citrus disease that is associated with bacteria of the genus 'Candidatus Liberibacter' (Ca. L.). Powerful diagnostic tools and management strategies are desired to control HLB. Host small RNAs (sRNA) play a vital role in regulating host responses to pathogen infection and are used as early diagnostic markers for many human diseases, including cancers. To determine whether citrus sRNAs regulate host responses to HLB, sRNAs were profiled from Citrus sinensis 10 and 14 weeks post grafting with Ca. L. asiaticus (Las)-positive or healthy tissue. Ten new microRNAs (miRNAs), 76 conserved miRNAs, and many small interfering RNAs (siRNAs) were discovered. Several miRNAs and siRNAs were highly induced by Las infection, and can be potentially developed into early diagnosis markers of HLB. miR399, which is induced by phosphorus starvation in other plant species, was induced specifically by infection of Las but not Spiroplasma citri that causes citrus stubborn-a disease with symptoms similar to HLB. We found a 35% reduction of phosphorus in Las-positive citrus trees compared to healthy trees. Applying phosphorus oxyanion solutions to HLB-positive sweet orange trees reduced HLB symptom severity and significantly improved fruit production during a 3-year field trial in south-west Florida. Our molecular, physiological, and field data suggest that phosphorus deficiency is linked to HLB disease symptomology.

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

Science.gov (United States)

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

2014-10-01

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

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

Science.gov (United States)

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

2016-09-15

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

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

Science.gov (United States)

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

2016-06-01

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

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

Directory of Open Access Journals (Sweden)

Isiordia-Aquino N, Ortiz-Catón M

2013-07-01

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

Molecular evolution of the actin-like MreB protein gene family in wall-less bacteria.

Science.gov (United States)

Ku, Chuan; Lo, Wen-Sui; Kuo, Chih-Horng

2014-04-18

The mreB gene family encodes actin-like proteins that determine cell shape by directing cell wall synthesis and often exists in one to three copies in the genomes of non-spherical bacteria. Intriguingly, while most wall-less bacteria do not have this gene, five to seven mreB homologs are found in Spiroplasma and Haloplasma, which are both characterized by cell contractility. To investigate the molecular evolution of this gene family in wall-less bacteria, we sampled the available genome sequences from these two genera and other related lineages for comparative analysis. The gene phylogenies indicated that the mreB homologs in Haloplasma are more closely related to those in Firmicutes, whereas those in Spiroplasma form a separate clade. This finding suggests that the gene family expansions in these two lineages are the results of independent ancient duplications. Moreover, the Spiroplasma mreB homologs can be classified into five clades, of which the genomic positions are largely conserved. The inference of gene gains and losses suggests that there has been an overall trend to retain only one homolog from each of the five mreB clades in the evolutionary history of Spiroplasma. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

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

Directory of Open Access Journals (Sweden)

Zhisheng Xie

2013-01-01

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

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

International Nuclear Information System (INIS)

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

2005-01-01

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

DNA polymorphisms and biocontrol of Bacillus antagonistic to citrus bacterial canker with indication of the interference of phyllosphere biofilms.

Directory of Open Access Journals (Sweden)

Tzu-Pi Huang

Full Text Available Citrus bacterial canker caused by Xanthomonas axonopodis pv. citri is a devastating disease resulting in significant crop losses in various citrus cultivars worldwide. A biocontrol agent has not been recommended for this disease. To explore the potential of bacilli native to Taiwan to control this disease, Bacillus species with a broad spectrum of antagonistic activity against various phytopathogens were isolated from plant potting mixes, organic compost and the rhizosphere soil. Seven strains TKS1-1, OF3-16, SP4-17, HSP1, WG6-14, TLB7-7, and WP8-12 showing superior antagonistic activity were chosen for biopesticide development. The genetic identity based on 16S rDNA sequences indicated that all seven native strains were close relatives of the B. subtilis group and appeared to be discrete from the B. cereus group. DNA polymorphisms in strains WG6-14, SP4-17, TKS1-1, and WP8-12, as revealed by repetitive sequence-based PCR with the BOXA1R primers were similar to each other, but different from those of the respective Bacillus type strains. However, molecular typing of the strains using either tDNA-intergenic spacer regions or 16S-23S intergenic transcribed spacer regions was unable to differentiate the strains at the species level. Strains TKS1-1 and WG6-14 attenuated symptom development of citrus bacterial canker, which was found to be correlated with a reduction in colonization and biofilm formation by X. axonopodis pv. citri on leaf surfaces. The application of a Bacillus strain TKS1-1 endospore formulation to the leaf surfaces of citrus reduced the incidence of citrus bacterial canker and could prevent development of the disease.

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

Science.gov (United States)

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

2018-04-24

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

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

Science.gov (United States)

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

2013-06-01

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

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. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

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

Science.gov (United States)

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

2003-07-01

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

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

Directory of Open Access Journals (Sweden)

Thomson M Paris

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

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

Science.gov (United States)

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

2016-12-01

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

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.

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

Directory of Open Access Journals (Sweden)

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.

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

International Nuclear Information System (INIS)

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

2012-01-01

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

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; Machado, Marcos Antonio

2017-01-01

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

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

Science.gov (United States)

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

2017-08-31

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

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

Science.gov (United States)

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

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

Directory of Open Access Journals (Sweden)

Diogo Manzano Galdeano

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

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

Science.gov (United States)

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

2018-01-01

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

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

KAUST Repository

Piazza, A.; Zimaro, T.; Garavaglia, B. S.; Ficarra, F. A.; Thomas, L.; Marondedze, C.; Feil, R.; Lunn, J. E.; Gehring, Christoph A; Ottado, J.; Gottig, N.

2015-01-01

and proteomic analyses of infected leaves showed that infection with XccΔotsA triggered only weak defence responses in the plant compared with infection with Xcc, and had less impact on the host plant's metabolism than the wild-type strain. These results

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

Science.gov (United States)

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

2018-03-23

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

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

Science.gov (United States)

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

2007-03-15

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

A veritable menagerie of heritable bacteria from ants, butterflies, and beyond: broad molecular surveys and a systematic review.

Directory of Open Access Journals (Sweden)

Jacob A Russell

Full Text Available Maternally transmitted bacteria have been important players in the evolution of insects and other arthropods, affecting their nutrition, defense, development, and reproduction. Wolbachia are the best studied among these and typically the most prevalent. While several other bacteria have independently evolved a heritable lifestyle, less is known about their host ranges. Moreover, most groups of insects have not had their heritable microflora systematically surveyed across a broad range of their taxonomic diversity. To help remedy these shortcomings we used diagnostic PCR to screen for five groups of heritable symbionts-Arsenophonus spp., Cardinium hertigii, Hamiltonella defensa, Spiroplasma spp., and Wolbachia spp.-across the ants and lepidopterans (focusing, in the latter case, on two butterfly families-the Lycaenidae and Nymphalidae. We did not detect Cardinium or Hamiltonella in any host. Wolbachia were the most widespread, while Spiroplasma (ants and lepidopterans and Arsenophonus (ants only were present at low levels. Co-infections with different Wolbachia strains appeared especially common in ants and less so in lepidopterans. While no additional facultative heritable symbionts were found among ants using universal bacterial primers, microbes related to heritable enteric bacteria were detected in several hosts. In summary, our findings show that Wolbachia are the dominant heritable symbionts of ants and at least some lepidopterans. However, a systematic review of symbiont frequencies across host taxa revealed that this is not always the case across other arthropods. Furthermore, comparisons of symbiont frequencies revealed that the prevalence of Wolbachia and other heritable symbionts varies substantially across lower-level arthropod taxa. We discuss the correlates, potential causes, and implications of these patterns, providing hypotheses on host attributes that may shape the distributions of these influential bacteria.

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

Directory of Open Access Journals (Sweden)

Kunming Qin

2013-01-01

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

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

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

Inhibition of Xanthomonas citri subsp. citri, causal agent of citrus canker, by a strain of Zymomonas mobilis mobilis isolated from sugarcane

OpenAIRE

Romero, María E.; Ramallo, C. Jacqueline; Ploper, L. Daniel

2008-01-01

Zymomonas mobilis mobilis (Zm) produce factores antimicrobianos que actúan sobre un amplio espectro de microorganismos patógenos para el hombre, animales y plantas. Un problema importante a resolver en los tratamientos con antimicrobianos, es el desarrollo de resistencia a compuestos empleados actualmente, no siendo las bacterias fitopatógenas una excepción. En el presente trabajo se realizaron ensayos de antagonismo con células (pruebas de estrías cruzadas) y sobrenadantes concentrados (Sc) ...

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

Science.gov (United States)

2011-01-27

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

Antibiotics from bacillus subtilis AECL69 8. isolation and purification of a complex of antibacterial antibiotics x

International Nuclear Information System (INIS)

Ahmad, M.S.; Malik, M.A.; Shaukat, G.A.

1996-01-01

A bacterial strain bacillus subtilis AECL69 produces two anti bacterial antibiotics in a specified complex or synthetic medium. One of the antibiotics is characteristically active against Xanthomonas citri. Procedures have been described to isolate and purify a complex of xanthmonas antibiotics from the fermented complex broths, and from the fermented synthetic medium as well. Paper chromatography coupled with bioautography has shown that the complex of xanthomonas antibiotics has at least three components. The three components were indicated irrespective of the fact whether it was isolated from the fermented complex or synthetic broth. (author)

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

Directory of Open Access Journals (Sweden)

Nabil Killiny

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

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

Science.gov (United States)

Killiny, Nabil; Jones, Shelley E

2018-01-01

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

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

Science.gov (United States)

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

2016-02-01

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

Ectopic Expression of Xylella fastidiosa rpfF Conferring Production of Diffusible Signal Factor in Transgenic Tobacco and Citrus Alters Pathogen Behavior and Reduces Disease Severity.

Science.gov (United States)

Caserta, R; Souza-Neto, R R; Takita, M A; Lindow, S E; De Souza, A A

2017-11-01

The pathogenicity of Xylella fastidiosa is associated with its ability to colonize the xylem of host plants. Expression of genes contributing to xylem colonization are suppressed, while those necessary for insect vector acquisition are increased with increasing concentrations of diffusible signal factor (DSF), whose production is dependent on RpfF. We previously demonstrated that transgenic citrus plants ectopically expressing rpfF from a citrus strain of X. fastidiosa subsp. pauca exhibited less susceptibility to Xanthomonas citri subsp. citri, another pathogen whose virulence is modulated by DSF accumulation. Here, we demonstrate that ectopic expression of rpfF in both transgenic tobacco and sweet orange also confers a reduction in disease severity incited by X. fastidiosa and reduces its colonization of those plants. Decreased disease severity in the transgenic plants was generally associated with increased expression of genes conferring adhesiveness to the pathogen and decreased expression of genes necessary for active motility, accounting for the reduced population sizes achieved in the plants, apparently by limiting pathogen dispersal through the plant. Plant-derived DSF signal molecules in a host plant can, therefore, be exploited to interfere with more than one pathogen whose virulence is controlled by DSF signaling.

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

Directory of Open Access Journals (Sweden)

Mamoudou Sétamou

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

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

Science.gov (United States)

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

2016-01-01

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

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

OpenAIRE

Killiny, Nabil

2016-01-01

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

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

Directory of Open Access Journals (Sweden)

Hofit Kol-Maimon

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

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

Science.gov (United States)

Chen, Yonggang; Lin, Li

2011-10-01

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

Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1.

Science.gov (United States)

Zhang, Shujian; Chakrabarty, Pranjib K; Fleites, Laura A; Rayside, Patricia A; Hopkins, Donald L; Gabriel, Dean W

2015-01-01

Xylella fastidiosa (X. fastidiosa) infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD) of grapevines. X. fastidiosa biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703) and a serine protease (PD0956); two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928), and at least one relatively short, hemagglutinin-like protein (PD0986). Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of E. coli BL21(DE3) overexpressing PD1703 exhibited a hypersensitive response (HR) in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to Xanthomonas citri, E. coli and X. fastidiosa EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot) and a PD0986 (hemagglutinin) were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on V. vinifera vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of X. fastidiosa populations in grapevines or elderberry.

Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1.

Directory of Open Access Journals (Sweden)

Shujian Zhang

Full Text Available Xylella fastidiosa (X. fastidiosa infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD of grapevines. X. fastidiosa biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703 and a serine protease (PD0956; two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928, and at least one relatively short, hemagglutinin-like protein (PD0986. Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of E. coli BL21(DE3 overexpressing PD1703 exhibited a hypersensitive response (HR in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to Xanthomonas citri, E. coli and X. fastidiosa EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot and a PD0986 (hemagglutinin were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on V. vinifera vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of X. fastidiosa populations in grapevines or elderberry.

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

Directory of Open Access Journals (Sweden)

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.

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

Science.gov (United States)

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

2015-10-01

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

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

Directory of Open Access Journals (Sweden)

Xuan Zeng

2018-04-01

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

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

Science.gov (United States)

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

A strain gauge

DEFF Research Database (Denmark)

2016-01-01

The invention relates to a strain gauge of a carrier layer and a meandering measurement grid positioned on the carrier layer, wherein the strain gauge comprises two reinforcement members positioned on the carrier layer at opposite ends of the measurement grid in the axial direction....... The reinforcement members are each placed within a certain axial distance to the measurement grid with the axial distance being equal to or smaller than a factor times the grid spacing. The invention further relates to a multi-axial strain gauge such as a bi-axial strain gauge or a strain gauge rosette where each...... of the strain gauges comprises reinforcement members. The invention further relates to a method for manufacturing a strain gauge as mentioned above....

Development of intra-strain self-cloning procedure for breeding baker's yeast strains.

Science.gov (United States)

Nakagawa, Youji; Ogihara, Hiroyuki; Mochizuki, Chisato; Yamamura, Hideki; Iimura, Yuzuru; Hayakawa, Masayuki

2017-03-01

Previously reported self-cloning procedures for breeding of industrial yeast strains require DNA from other strains, plasmid DNA, or mutagenesis. Therefore, we aimed to construct a self-cloning baker's yeast strain that exhibits freeze tolerance via an improved self-cloning procedure. We first disrupted the URA3 gene of a prototrophic baker's yeast strain without the use of any marker gene, resulting in a Δura3 homozygous disruptant. Then, the URA3 gene of the parental baker's yeast strain was used as a selection marker to introduce the constitutive TDH3 promoter upstream of the PDE2 gene encoding high-affinity cyclic AMP phosphodiesterase. This self-cloning procedure was performed without using DNA from other Saccharomyces cerevisiae strains, plasmid DNA, or mutagenesis and was therefore designated an intra-strain self-cloning procedure. Using this self-cloning procedure, we succeeded in producing self-cloning baker's yeast strains that harbor the TDH3p-PDE2 gene heterozygously and homozygously, designated TDH3p-PDE2 hetero and TDH3p-PDE2 homo strains, respectively. These self-cloning strains expressed much higher levels of PDE2 mRNA than the parental strain and exhibited higher viability after freeze stress, as well as higher fermentation ability in frozen dough, when compared with the parental strain. The TDH3p-PDE2 homo strain was genetically more stable than the TDH3p-PDE2 hetero strain. These results indicate that both heterozygous and homozygous strains of self-cloning PDE2-overexpressing freeze-tolerant strains of industrial baker's yeast can be prepared using the intra-strain self-cloning procedure, and, from a practical viewpoint, the TDH3p-PDE2 homo strain constructed in this study is preferable to the TDH3p-PDE2 hetero strain for frozen dough baking. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

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

Directory of Open Access Journals (Sweden)

Everth Emilio Ebratt Ravelo

2011-12-01

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

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

Strain hardening rate sensitivity and strain rate sensitivity in TWIP steels

Energy Technology Data Exchange (ETDEWEB)

Bintu, Alexandra [TEMA, Department of Mechanical Engineering, University of Aveiro, Campus Universitário de Santiago, 3810-193 (Portugal); Vincze, Gabriela, E-mail: gvincze@ua.pt [TEMA, Department of Mechanical Engineering, University of Aveiro, Campus Universitário de Santiago, 3810-193 (Portugal); Picu, Catalin R. [Department of Mechanical, Aerospace and Nuclear Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180 (United States); Lopes, Augusto B. [CICECO, Department of Materials and Ceramic Engineering, University of Aveiro, Campus Universitário de Santiago, 3810-193 (Portugal); Grácio, Jose J. [TEMA, Department of Mechanical Engineering, University of Aveiro, Campus Universitário de Santiago, 3810-193 (Portugal); Barlat, Frederic [Materials Mechanics Laboratory, Graduate Institute of Ferrous Technology, Pohang University of Science and Technology, Pohang 790-784 (Korea, Republic of)

2015-04-01

TWIP steels are materials with very high strength and exceptional strain hardening capability, parameters leading to large energy absorption before failure. However, TWIP steels also exhibit reduced (often negative) strain rate sensitivity (SRS) which limits the post-necking deformation. In this study we demonstrate for an austenitic TWIP steel with 18% Mn a strong dependence of the twinning rate on the strain rate, which results in negative strain hardening rate sensitivity (SHRS). The instantaneous component of SHRS is large and negative, while its transient is close to zero. The SRS is observed to decrease with strain, becoming negative for larger strains. Direct observations of the strain rate dependence of the twinning rate are made using electron microscopy and electron backscatter diffraction, which substantiate the proposed mechanism for the observed negative SHRS.

Strain hardening rate sensitivity and strain rate sensitivity in TWIP steels

International Nuclear Information System (INIS)

Bintu, Alexandra; Vincze, Gabriela; Picu, Catalin R.; Lopes, Augusto B.; Grácio, Jose J.; Barlat, Frederic

2015-01-01

TWIP steels are materials with very high strength and exceptional strain hardening capability, parameters leading to large energy absorption before failure. However, TWIP steels also exhibit reduced (often negative) strain rate sensitivity (SRS) which limits the post-necking deformation. In this study we demonstrate for an austenitic TWIP steel with 18% Mn a strong dependence of the twinning rate on the strain rate, which results in negative strain hardening rate sensitivity (SHRS). The instantaneous component of SHRS is large and negative, while its transient is close to zero. The SRS is observed to decrease with strain, becoming negative for larger strains. Direct observations of the strain rate dependence of the twinning rate are made using electron microscopy and electron backscatter diffraction, which substantiate the proposed mechanism for the observed negative SHRS

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

Science.gov (United States)

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

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

Science.gov (United States)

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

Variation in the strain anisotropy of Zircaloy with temperature and strain

International Nuclear Information System (INIS)

Hindle, E.D.; Worswick, D.

1984-04-01

Strain anisotropy was investigated at temperatures in the range 293 to 1117K in circular tensile specimens prepared from rolled Zircaloy-2 plate so that their tensile axes were parallel to and transverse to the rolling direction. The strain anisotropy factor for both types of specimen increased markedly in the high alpha phase region above 923K reaching a maximum at circa 1070K. Above this temperature in the alpha-plus-beta phase region the strain anisotropy decreased rapidly as the proportion of beta phase increased and was almost non-existent at 1173K. The strain anisotropy was markedly strain dependent, particularly in the high alpha phase region. The study was extended to Zircaloy-4 pressurized water reactor (PWR) 17 x 17 type fuel rod tubing specimens which were strained under biaxial conditions using cooling conditions which promoted uniform diametral strain over most of their lengths (circa 250 mm). In these circumstances the strain anisotropy is manifest by a reduction in length. Measurement of this change along with that in diameter and wall thickness produced data from which the strain anisotropy factor was calculated. The results, although influenced by additional factors discussed in the paper, were similar to those observed in the uniaxial Zircaloy-2 tensile tests. (author)

Unrelated facultative endosymbionts protect aphids against a fungal pathogen.

Science.gov (United States)

Łukasik, Piotr; van Asch, Margriet; Guo, Huifang; Ferrari, Julia; Godfray, H Charles J

2013-02-01

The importance of microbial facultative endosymbionts to insects is increasingly being recognized, but our understanding of how the fitness effects of infection are distributed across symbiont taxa is limited. In the pea aphid, some of the seven known species of facultative symbionts influence their host's resistance to natural enemies, including parasitoid wasps and a pathogenic fungus. Here we show that protection against this entomopathogen, Pandora neoaphidis, can be conferred by strains of four distantly related symbionts (in the genera Regiella, Rickettsia, Rickettsiella and Spiroplasma). They reduce mortality and also decrease fungal sporulation on dead aphids which may help protect nearby genetically identical insects. Pea aphids thus obtain protection from natural enemies through association with a wider range of microbial associates than has previously been thought. Providing resistance against natural enemies appears to be a particularly common way for facultative endosymbionts to increase in frequency within host populations. © 2012 Blackwell Publishing Ltd/CNRS.

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

Science.gov (United States)

2013-02-06

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

Haemophilus ducreyi Cutaneous Ulcer Strains Are Nearly Identical to Class I Genital Ulcer Strains.

Directory of Open Access Journals (Sweden)

Dharanesh Gangaiah

Full Text Available Although cutaneous ulcers (CU in the tropics is frequently attributed to Treponema pallidum subspecies pertenue, the causative agent of yaws, Haemophilus ducreyi has emerged as a major cause of CU in yaws-endemic regions of the South Pacific islands and Africa. H. ducreyi is generally susceptible to macrolides, but CU strains persist after mass drug administration of azithromycin for yaws or trachoma. H. ducreyi also causes genital ulcers (GU and was thought to be exclusively transmitted by microabrasions that occur during sex. In human volunteers, the GU strain 35000HP does not infect intact skin; wounds are required to initiate infection. These data led to several questions: Are CU strains a new variant of H. ducreyi or did they evolve from GU strains? Do CU strains contain additional genes that could allow them to infect intact skin? Are CU strains susceptible to azithromycin?To address these questions, we performed whole-genome sequencing and antibiotic susceptibility testing of 5 CU strains obtained from Samoa and Vanuatu and 9 archived class I and class II GU strains. Except for single nucleotide polymorphisms, the CU strains were genetically almost identical to the class I strain 35000HP and had no additional genetic content. Phylogenetic analysis showed that class I and class II strains formed two separate clusters and CU strains evolved from class I strains. Class I strains diverged from class II strains ~1.95 million years ago (mya and CU strains diverged from the class I strain 35000HP ~0.18 mya. CU and GU strains evolved under similar selection pressures. Like 35000HP, the CU strains were highly susceptible to antibiotics, including azithromycin.These data suggest that CU strains are derivatives of class I strains that were not recognized until recently. These findings require confirmation by analysis of CU strains from other regions.

Factors affecting finite strain estimation in low-grade, low-strain clastic rocks

Science.gov (United States)

Pastor-Galán, Daniel; Gutiérrez-Alonso, Gabriel; Meere, Patrick A.; Mulchrone, Kieran F.

2009-12-01

The computer strain analysis methods SAPE, MRL and DTNNM have permitted the characterization of finite strain in two different regions with contrasting geodynamic scenarios; (1) the Talas Ala Tau (Tien Shan, Kyrgyzs Republic) and (2) the Somiedo Nappe and Narcea Antiform (Cantabrian to West Asturian-Leonese Zone boundary, Variscan Belt, NW of Iberia). The performed analyses have revealed low-strain values and the regional strain trend in both studied areas. This study also investigates the relationship between lithology (grain size and percentage of matrix) and strain estimates the two methodologies used. The results show that these methods are comparable and the absence of significant finite strain lithological control in rocks deformed under low metamorphic and low-strain conditions.

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.

Combined Creutzfeldt-Jakob/ Alzheimer's Disease Cases are Important in Search for Microbes in Alzheimer's Disease.

Science.gov (United States)

Bastian, Frank O

2017-01-01

The question whether Alzheimer's disease is infectious as brought up in the recent editorial published in the Journal of Alzheimer's Disease is complicated by the controversy whether the causal agent is a microbe or a misfolded host protein (amyloid). The replicating amyloid (prion) theory, based upon data from studies of Creutzfeldt-Jakob disease (CJD) and other transmissible spongiform encephalopathies (TSEs), has been challenged since the prion can be separated from TSE infectivity, and spiroplasma, a wall-less bacterium, has been shown to be involved in the pathogenesis of CJD. Further support for a microbial cause for AD comes from occurrence of mixed CJD/AD cases involving up to 15% of AD brains submitted to brain banks. The association of CJD with AD suggests a common etiology rather than simply being a medical curiosity. A co-infection with the transmissible agent of CJD, which we propose to be a Spiroplasma sp., would explain the diversity of bacteria shown to be associated with cases of AD.

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

Directory of Open Access Journals (Sweden)

Edvan Alves Chagas

2007-07-01

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

Chemicalome and metabolome profiling of polymethoxylated flavonoids in Citri Reticulatae Pericarpium based on an integrated strategy combining background subtraction and modified mass defect filter in a Microsoft Excel Platform.

Science.gov (United States)

Zeng, Su-Ling; Duan, Li; Chen, Bai-Zhong; Li, Ping; Liu, E-Hu

2017-07-28

Detection of metabolites in complex biological matrixes is a great challenge because of the background noise and endogenous components. Herein, we proposed an integrated strategy that combined background subtraction program and modified mass defect filter (MMDF) data mining in a Microsoft Excel platform for chemicalome and metabolome profiling of the polymethoxylated flavonoids (PMFs) in Citri Reticulatae Pericarpium (CRP). The exogenously-sourced ions were firstly filtered out by the developed Visual Basic for Applications (VBA) program incorporated in the Microsoft Office. The novel MMDF strategy was proposed for detecting both target and untarget constituents and metabolites based on narrow, well-defined mass defect ranges. The approach was validated to be powerful, and potentially useful for the metabolite identification of both single compound and homologous compound mixture. We successfully identified 30 and 31 metabolites from rat biosamples after oral administration of nobiletin and tangeretin, respectively. A total of 56 PMFs compounds were chemically characterized and 125 metabolites were captured. This work demonstrated the feasibility of the integrated approach for reliable characterization of the constituents and metabolites in herbal medicines. Copyright © 2017 Elsevier B.V. All rights reserved.

Evaluation of Antimicrobial Activity of Ginger (Zingiber officinale) Essential Oil Treated by Gamma Radiation

International Nuclear Information System (INIS)

Abdeldaiem, M.H.; Helal, I.M.

2009-01-01

The essential oil of ginger (Zingiber officinale) was irradiated at doses of 0,5, 10 and 15 kGy. The irradiated as well as non-irradiated oil were tested for their antibacterial activity against one strain of food poisoning bacteria (Bacillus cereus), one strain of the indicator microorganisms (Escherichia coli), two strains of human pathogenic bacteria (Klebsiella pneumonia, and Pseudomonas aeruginosa), one strain of food spoilage bacteria (Bacillus subtilis) and one strain of plant pathogenic bacteria(Pseudomonas citri). In addition, the studied oils were tested for their antifungal activities against five strains of plant pathogenic fungi (Aspergillus niger, Aspergillus flavus, Fusarium oxysporium, Trichoderma viride and Alternaria alternata) to investigate the effect of gamma irradiation on the antimicrobial activity of the tested oil. The obtained results showed that all tested oils exhibited an inhibition effect for the growth of the microorganisms under investigation. While, Trichoderma viride appeared to be the most resistant fungus with non-irradiated and irradiated ginger essential oil treatments. The ginger essential oil irradiated at dose of 10 kGy appeared to be the best one for inhibition the growth of most tested bacteria and fungi under investigation. Also, changes in chemical constituents of ginger essential oil as affected by gamma radiation were investigated by Gas Chromatography

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

Directory of Open Access Journals (Sweden)

Justin George

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

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

Science.gov (United States)

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

2017-01-01

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

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.

Stress-strain properties of railway steel at strain rates of upto 105 per second

International Nuclear Information System (INIS)

Hashmi, M.S.J.; Islam, M.N.

1985-01-01

This paper presents the stress-strain characteristics of railway steel at strain rates of up to 10 5 /s at room temperature determined by a new technique. In determining the results, account has been taken of the strain-rate variation, the total strain and the strain rate history. The effect of friction, material inertia and temperature rise is also assessed and an empirical constitutive equation describing the strain-rate and strain sensitive flow stress for this type of steel is proposed. (orig.)

Comparison of Thermal Creep Strain Calculation Results Using Time Hardening and Strain Hardening Rules

International Nuclear Information System (INIS)

Kim, Junehyung; Cheon, Jinsik; Lee, Byoungoon; Lee, Chanbock

2014-01-01

One of the design criteria for the fuel rod in PGSFR is the thermal creep strain of the cladding, because the cladding is exposed to a high temperature for a long time during reactor operation period. In general, there are two kind of calculation scheme for thermal creep strain: time hardening and strain hardening rules. In this work, thermal creep strain calculation results for HT9 cladding by using time hardening and strain hardening rules are compared by employing KAERI's current metallic fuel performance analysis code, MACSIS. Also, thermal creep strain calculation results by using ANL's metallic fuel performance analysis code, LIFE-METAL which adopts strain hardening rule are compared with those by using MACSIS. Thermal creep strain calculation results for HT9 cladding by using time hardening and strain hardening rules were compared by employing KAERI's current metallic fuel performance analysis code, MACSIS. Also, thermal creep strain calculation results by using ANL's metallic fuel performance analysis code, LIFE-METAL which adopts strain hardening rule were compared with those by using MACSIS. Tertiary creep started earlier in time hardening rule than in strain hardening rule. Also, calculation results by MACSIS with strain hardening and those obtained by using LIFE-METAL were almost identical to each other

Limitations of Hollomon and Ludwigson stress-strain relations in assessing the strain hardening parameters

International Nuclear Information System (INIS)

Samuel, K G

2006-01-01

It is shown that the deviation from the ideal Hollomon relation in describing the stress-strain behaviour is characteristic of all materials at low strains. The Ludwigson relation describing the deviation from the Hollomon relation at low strains is critically analysed and it is shown that the deviation at low strains is a consequence of some unknown 'plastic strain equivalent' present in the material. Stress strain curves obeying an ideal Hollomon relation as well as that of a structurally modified (prior cold worked) material were simulated and compared. The results show that the yield strength and the flow strength of a material at constant strain rate and temperature are dictated by the magnitude of the 'plastic strain equivalent' term. It is shown that this component need not necessarily mean a prior plastic strain present in the material due to prior cold work alone and that prior cold work strain will add to this. If this component is identified, the stress-strain behaviour can be adequately described by the Swift relation. It is shown that in both formalisms, the strain hardening index is a function of the yield strength of the material

Simultaneous Quantification of Seven Bioactive Flavonoids in Citri Reticulatae Pericarpium by Ultra-Fast Liquid Chromatography Coupled with Tandem Mass Spectrometry.

Science.gov (United States)

Zhao, Lian-Hua; Zhao, Hong-Zheng; Zhao, Xue; Kong, Wei-Jun; Hu, Yi-Chen; Yang, Shi-Hai; Yang, Mei-Hua

2016-05-01

Citri Reticulatae Pericarpium (CRP) is a commonly-used traditional Chinese medicine with flavonoids as the major bioactive components. Nevertheless, the contents of the flavonoids in CRP of different sources may significantly vary affecting their therapeutic effects. Thus, the setting up of a reliable and comprehensive quality assessment method for flavonoids in CRP is necessary. To set up a rapid and sensitive ultra-fast liquid chromatography coupled with tandem mass spectrometry (UFLC-MS/MS) method for simultaneous quantification of seven bioactive flavonoids in CRP. A UFLC-MS/MS method coupled to ultrasound-assisted extraction was developed for simultaneous separation and quantification of seven flavonoids including hesperidin, neohesperidin, naringin, narirutin, tangeretin, nobiletin and sinensetin in 16 batches of CRP samples from different sources in China. The established method showed good linearity for all analytes with correlation coefficient (R) over 0.9980, together with satisfactory accuracy, precision and reproducibility. Furthermore, the recoveries at the three spiked levels were higher than 89.71% with relative standard deviations (RSDs) lower than 5.19%. The results indicated that the contents of seven bioactive flavonoids in CRP varied significantly among different sources. Among the samples under study, hesperidin showed the highest contents in 16 samples ranged from 27.50 to 86.30 mg/g, the contents of hesperidin in CRP-15 and CRP-9 were 27.50 and 86.30 mg/g, respectively, while, the amount of narirutin was too low to be measured in some samples. This study revealed that the developed UFLC-MS/MS method was simple, sensitive and reliable for simultaneous quantification of multi-components in CRP with potential perspective for quality control of complex matrices. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Strained Silicon Photonics

Directory of Open Access Journals (Sweden)

Ralf B. Wehrspohn

2012-05-01

Full Text Available A review of recent progress in the field of strained silicon photonics is presented. The application of strain to waveguide and photonic crystal structures can be used to alter the linear and nonlinear optical properties of these devices. Here, methods for the fabrication of strained devices are summarized and recent examples of linear and nonlinear optical devices are discussed. Furthermore, the relation between strain and the enhancement of the second order nonlinear susceptibility is investigated, which may enable the construction of optically active photonic devices made of silicon.

Amerindian Helicobacter pylori strains go extinct, as european strains expand their host range.

Directory of Open Access Journals (Sweden)

Maria G Domínguez-Bello

Full Text Available We studied the diversity of bacteria and host in the H. pylori-human model. The human indigenous bacterium H. pylori diverged along with humans, into African, European, Asian and Amerindian groups. Of these, Amerindians have the least genetic diversity. Since niche diversity widens the sets of resources for colonizing species, we predicted that the Amerindian H. pylori strains would be the least diverse. We analyzed the multilocus sequence (7 housekeeping genes of 131 strains: 19 cultured from Africans, 36 from Spanish, 11 from Koreans, 43 from Amerindians and 22 from South American Mestizos. We found that all strains that had been cultured from Africans were African strains (hpAfrica1, all from Spanish were European (hpEurope and all from Koreans were hspEAsia but that Amerindians and Mestizos carried mixed strains: hspAmerind and hpEurope strains had been cultured from Amerindians and hpEurope and hpAfrica1 were cultured from Mestizos. The least genetically diverse H. pylori strains were hspAmerind. Strains hpEurope were the most diverse and showed remarkable multilocus sequence mosaicism (indicating recombination. The lower genetic structure in hpEurope strains is consistent with colonization of a diversity of hosts. If diversity is important for the success of H. pylori, then the low diversity of Amerindian strains might be linked to their apparent tendency to disappear. This suggests that Amerindian strains may lack the needed diversity to survive the diversity brought by non-Amerindian hosts.

POTENTIAL DISTRIBUTION OF ORANGE JASMINE (Murraya paniculata IN MEXICO

Directory of Open Access Journals (Sweden)

José López-Collado

2013-04-01

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

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

KAUST Repository

Garavaglia, Betiana S; Thomas, Ludivine; Zimaro, Tamara; Gottig, Natalia; Daurelio, Lucas D; Ndimba, Bongani; Orellano, Elena G; Ottado, Jorgelina; Gehring, Christoph A

2010-01-01

Background: Plant natriuretic peptides (PNPs) belong to a novel class of peptidic signaling molecules that share some structural similarity to the N-terminal domain of expansins and affect physiological processes such as water and ion homeostasis at nano-molar concentrations. The citrus pathogen Xanthomonas axonopodis pv. citri possesses a PNP-like peptide (XacPNP) uniquely present in this bacteria. Previously we observed that the expression of XacPNP is induced upon infection and that lesions produced in leaves infected with a XacPNP deletion mutant were more necrotic and lead to earlier bacterial cell death, suggesting that the plant-like bacterial PNP enables the plant pathogen to modify host responses in order to create conditions favorable to its own survival.Results: Here we measured chlorophyll fluorescence parameters and water potential of citrus leaves infiltrated with recombinant purified XacPNP and demonstrate that the peptide improves the physiological conditions of the tissue. Importantly, the proteomic analysis revealed that these responses are mirrored by rapid changes in the host proteome that include the up-regulation of Rubisco activase, ATP synthase CF1 ? subunit, maturase K, and ?- and ?-tubulin.Conclusions: We demonstrate that XacPNP induces changes in host photosynthesis at the level of protein expression and in photosynthetic efficiency in particular. Our findings suggest that the biotrophic pathogen can use the plant-like hormone to modulate the host cellular environment and in particular host metabolism and that such modulations weaken host defence. 2010 Garavaglia et al; licensee BioMed Central Ltd.

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

KAUST Repository

Garavaglia, Betiana S

2010-03-21

Background: Plant natriuretic peptides (PNPs) belong to a novel class of peptidic signaling molecules that share some structural similarity to the N-terminal domain of expansins and affect physiological processes such as water and ion homeostasis at nano-molar concentrations. The citrus pathogen Xanthomonas axonopodis pv. citri possesses a PNP-like peptide (XacPNP) uniquely present in this bacteria. Previously we observed that the expression of XacPNP is induced upon infection and that lesions produced in leaves infected with a XacPNP deletion mutant were more necrotic and lead to earlier bacterial cell death, suggesting that the plant-like bacterial PNP enables the plant pathogen to modify host responses in order to create conditions favorable to its own survival.Results: Here we measured chlorophyll fluorescence parameters and water potential of citrus leaves infiltrated with recombinant purified XacPNP and demonstrate that the peptide improves the physiological conditions of the tissue. Importantly, the proteomic analysis revealed that these responses are mirrored by rapid changes in the host proteome that include the up-regulation of Rubisco activase, ATP synthase CF1 ? subunit, maturase K, and ?- and ?-tubulin.Conclusions: We demonstrate that XacPNP induces changes in host photosynthesis at the level of protein expression and in photosynthetic efficiency in particular. Our findings suggest that the biotrophic pathogen can use the plant-like hormone to modulate the host cellular environment and in particular host metabolism and that such modulations weaken host defence. 2010 Garavaglia et al; licensee BioMed Central Ltd.

Measurement of Strain and Strain Rate during the Impact of Tennis Ball Cores

Directory of Open Access Journals (Sweden)

Ben Lane

2018-03-01

Full Text Available The aim of this investigation was to establish the strains and strain rates experienced by tennis ball cores during impact to inform material characterisation testing and finite element modelling. Three-dimensional surface strains and strain rates were measured using two high-speed video cameras and corresponding digital image correlation software (GOM Correlate Professional. The results suggest that material characterisation testing to a maximum strain of 0.4 and a maximum rate of 500 s−1 in tension and to a maximum strain of −0.4 and a maximum rate of −800 s−1 in compression would encapsulate the demands placed on the material during impact and, in turn, define the range of properties required to encapsulate the behavior of the material during impact, enabling testing to be application-specific and strain-rate-dependent properties to be established and incorporated in finite element models.

Strain measurement based battery testing

Science.gov (United States)

Xu, Jeff Qiang; Steiber, Joe; Wall, Craig M.; Smith, Robert; Ng, Cheuk

2017-05-23

A method and system for strain-based estimation of the state of health of a battery, from an initial state to an aged state, is provided. A strain gauge is applied to the battery. A first strain measurement is performed on the battery, using the strain gauge, at a selected charge capacity of the battery and at the initial state of the battery. A second strain measurement is performed on the battery, using the strain gauge, at the selected charge capacity of the battery and at the aged state of the battery. The capacity degradation of the battery is estimated as the difference between the first and second strain measurements divided by the first strain measurement.

Strain measurement technique

International Nuclear Information System (INIS)

1987-01-01

The 10 contributions are concerned with selected areas of application, such as strain measurements in wood, rubber/metal compounds, sets of strain measurements on buildings, reinforced concrete structures without gaps, pipes buried in the ground and measurements of pressure fluctuations. To increase the availability and safety of plant, stress analyses were made on gas turbine rotors with HT-DMS or capacitive HT-DMS (high temperature strain measurements). (DG) [de

Strain expansion-reduction approach

Science.gov (United States)

Baqersad, Javad; Bharadwaj, Kedar

2018-02-01

Validating numerical models are one of the main aspects of engineering design. However, correlating million degrees of freedom of numerical models to the few degrees of freedom of test models is challenging. Reduction/expansion approaches have been traditionally used to match these degrees of freedom. However, the conventional reduction/expansion approaches are only limited to displacement, velocity or acceleration data. While in many cases only strain data are accessible (e.g. when a structure is monitored using strain-gages), the conventional approaches are not capable of expanding strain data. To bridge this gap, the current paper outlines a reduction/expansion technique to reduce/expand strain data. In the proposed approach, strain mode shapes of a structure are extracted using the finite element method or the digital image correlation technique. The strain mode shapes are used to generate a transformation matrix that can expand the limited set of measurement data. The proposed approach can be used to correlate experimental and analytical strain data. Furthermore, the proposed technique can be used to expand real-time operating data for structural health monitoring (SHM). In order to verify the accuracy of the approach, the proposed technique was used to expand the limited set of real-time operating data in a numerical model of a cantilever beam subjected to various types of excitations. The proposed technique was also applied to expand real-time operating data measured using a few strain gages mounted to an aluminum beam. It was shown that the proposed approach can effectively expand the strain data at limited locations to accurately predict the strain at locations where no sensors were placed.

Citri Reticulatae Pericarpium (Chenpi): Botany, ethnopharmacology, phytochemistry, and pharmacology of a frequently used traditional Chinese medicine.

Science.gov (United States)

Yu, Xin; Sun, Shuang; Guo, Yuyan; Liu, Yan; Yang, Dayu; Li, Guoyu; Lü, Shaowa

2018-06-28

Citri Reticulatae Pericarpium (Rutaceae, CRP), commonly called as Chenpi () in Chinese, is most frequently used as a qi-regulating drug in thousands of Chinese medicine prescriptions. CRP is found mainly in major citrus-producing areas such as the Guangdong, Guangxi, Sichuan, Fujian, and Zhejiang Provinces of China. Since thousands of years in China, CRP has been used widely in clinical practice to treat nausea, vomiting, indigestion, anepithymia, diarrhea, cough, expectoration, and so on. Currently, CRP is listed in the Pharmacopoeia of the People's Republic of China. The present paper reviews the botany, ethnopharmacology, phytochemistry, pharmacology, quality control, and toxicology of CRP. Information on CRP was gathered from various sources including the books on traditional Chinese herbal medicine; scientific databases including Elsevier, PubMed, and ScienceDirect; Baidu Scholar; CNKI; and others and from different professional websites. Approximately 140 chemical compounds have been isolated and identified from CRP. Among them, volatile oils and flavonoids are generally considered as the main bioactive and characteristic ingredients. CRP possesses wide pharmacological effects such as having a beneficial effect on the cardiovascular, digestive, and respiratory systems, antitumor, antioxidant, and anti-inflammatory properties; and a protective effect on the liver and nerve. Moreover, hesperidin is chosen as an indicator in the quantitative determination of CRP, and the quantity of aflatoxin in CRP must not exceed the standard limit mentioned in the pharmacopoeia. In brief, CRP has a warming nature, and hence, it can be used in harmony with a lot of medicines. CRP not only exhibits its effects individually but also aids other medicines exhibit a better effect. CRP can be consumed with tea, food, alcohol, and medicine. Irrespective of the form it is being consumed, CRP not only shows a synergistic effect but also has strengths on its own. Modern pharmacological

Rat Strain Ontology: structured controlled vocabulary designed to facilitate access to strain data at RGD.

Science.gov (United States)

Nigam, Rajni; Munzenmaier, Diane H; Worthey, Elizabeth A; Dwinell, Melinda R; Shimoyama, Mary; Jacob, Howard J

2013-11-22

The Rat Genome Database (RGD) ( http://rgd.mcw.edu/) is the premier site for comprehensive data on the different strains of the laboratory rat (Rattus norvegicus). The strain data are collected from various publications, direct submissions from individual researchers, and rat providers worldwide. Rat strain, substrain designation and nomenclature follow the Guidelines for Nomenclature of Mouse and Rat Strains, instituted by the International Committee on Standardized Genetic Nomenclature for Mice. While symbols and names aid in identifying strains correctly, the flat nature of this information prohibits easy search and retrieval, as well as other data mining functions. In order to improve these functionalities, particularly in ontology-based tools, the Rat Strain Ontology (RS) was developed. The Rat Strain Ontology (RS) reflects the breeding history, parental background, and genetic manipulation of rat strains. This controlled vocabulary organizes strains by type: inbred, outbred, chromosome altered, congenic, mutant and so on. In addition, under the chromosome altered category, strains are organized by chromosome, and further by type of manipulations, such as mutant or congenic. This allows users to easily retrieve strains of interest with modifications in specific genomic regions. The ontology was developed using the Open Biological and Biomedical Ontology (OBO) file format, and is organized on the Directed Acyclic Graph (DAG) structure. Rat Strain Ontology IDs are included as part of the strain report (RS: ######). As rat researchers are often unaware of the number of substrains or altered strains within a breeding line, this vocabulary now provides an easy way to retrieve all substrains and accompanying information. Its usefulness is particularly evident in tools such as the PhenoMiner at RGD, where users can now easily retrieve phenotype measurement data for related strains, strains with similar backgrounds or those with similar introgressed regions. This

Inducible expression of Bs2 R gene from Capsicum chacoense in sweet orange (Citrus sinensis L. Osbeck) confers enhanced resistance to citrus canker disease.

Science.gov (United States)

Sendín, Lorena Noelia; Orce, Ingrid Georgina; Gómez, Rocío Liliana; Enrique, Ramón; Grellet Bournonville, Carlos Froilán; Noguera, Aldo Sergio; Vojnov, Adrián Alberto; Marano, María Rosa; Castagnaro, Atilio Pedro; Filippone, María Paula

2017-04-01

Transgenic expression of the pepper Bs2 gene confers resistance to Xanthomonas campestris pv. vesicatoria (Xcv) pathogenic strains which contain the avrBs2 avirulence gene in susceptible pepper and tomato varieties. The avrBs2 gene is highly conserved among members of the Xanthomonas genus, and the avrBs2 of Xcv shares 96% homology with the avrBs2 of Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker disease. A previous study showed that the transient expression of pepper Bs2 in lemon leaves reduced canker formation and induced plant defence mechanisms. In this work, the effect of the stable expression of Bs2 gene on citrus canker resistance was evaluated in transgenic plants of Citrus sinensis cv. Pineapple. Interestingly, Agrobacterium-mediated transformation of epicotyls was unsuccessful when a constitutive promoter (2× CaMV 35S) was used in the plasmid construction, but seven transgenic lines were obtained with a genetic construction harbouring Bs2 under the control of a pathogen-inducible promoter, from glutathione S-transferase gene from potato. A reduction of disease symptoms of up to 70% was observed in transgenic lines expressing Bs2 with respect to non-transformed control plants. This reduction was directly dependent on the Xcc avrBs2 gene since no effect was observed when a mutant strain of Xcc with a disruption in avrBs2 gene was used for inoculations. Additionally, a canker symptom reduction was correlated with levels of the Bs2 expression in transgenic plants, as assessed by real-time qPCR, and accompanied by the production of reactive oxygen species. These results indicate that the pepper Bs2 resistance gene is also functional in a family other than the Solanaceae, and could be considered for canker control.

Variation in the strain anisotropy of Zircaloy with temperature and strain

International Nuclear Information System (INIS)

Hindle, E.D.; Worswick, D.

1984-01-01

The strong crystallographic texture which is developed during the fabrication of zirconium-based alloys causes pronounced anisotropy in their mechanical properties, particularly deformation. The tendency for circular-section tension specimens with a high concentration of basal poles in one direction to become elliptical when deformed in tension has been used in this study to provide quantitative data on the effects of both strain and temperature on strain anisotropy. Tension tests were carried out over a temperature range of 293 to 1193 K on specimens machined from Zircaloy-2 plate. The strain anisotropy was found to increase markedly at temperatures over 923 K, reaching a maximum in the region of 1070 K. The strain anisotropy increased with increasing strain in this temperature region. The study was extended to Zircaloy-4 pressurized-water reactor fuel cladding by carrying out tube swelling tests and evaluating the axial deformation produced. Although scatter in the test results was higher than that exhibited in the tension tests, the general trend in the data was similar. The effects of the strain anisotropy observed are discussed in relation to the effects of temperature on the ductility of Zircaloy fuel cladding tubes during postulated largebreak loss-of-coolant accidents

The effect of pre spring spray to reduce of citrus important pests.

Science.gov (United States)

Hashemi, B; Damavandian, M R; Shoushtaril, R Vafaei; Tafaghodynia, B

2008-10-01

The importance of pre spring spray against citrus aphids, Pulvinaria aurantii Cockerell and Panonychus citri McGregor that are the most important pest of citrus during spring was tested. In this research, 150 trees ten years old sweet orange (Thomson navel on Citrus aurantium (root stocks)) in a citrus orchard approximately three hectares sampled. The experiment was laid out in a totally randomized (one-way) design replicated five times. According to the results, the pre spring spray do not effect on population density of citrus aphids and P. aurantii during March, April, May and June. However, the P. citri population decreased. Therefore, it seems the pre spring spray in citrus orchards is not necessary, but if P. citri is observed, the pre spring spray should be recommended.

Three dimensional strained semiconductors

Science.gov (United States)

Voss, Lars; Conway, Adam; Nikolic, Rebecca J.; Leao, Cedric Rocha; Shao, Qinghui

2016-11-08

In one embodiment, an apparatus includes a three dimensional structure comprising a semiconductor material, and at least one thin film in contact with at least one exterior surface of the three dimensional structure for inducing a strain in the structure, the thin film being characterized as providing at least one of: an induced strain of at least 0.05%, and an induced strain in at least 5% of a volume of the three dimensional structure. In another embodiment, a method includes forming a three dimensional structure comprising a semiconductor material, and depositing at least one thin film on at least one surface of the three dimensional structure for inducing a strain in the structure, the thin film being characterized as providing at least one of: an induced strain of at least 0.05%, and an induced strain in at least 5% of a volume of the structure.

Hamstring strain - aftercare

Science.gov (United States)

Pulled hamstring muscle; Sprain - hamstring ... There are 3 levels of hamstring strains: Grade 1 -- mild muscle strain or pull Grade 2 -- partial muscle tear Grade 3 -- complete muscle tear Recovery time depends ...

Strain-Modulated Epitaxy

National Research Council Canada - National Science Library

Brown, April

1999-01-01

Strain-Modulated Epitaxy (SME) is a novel approach, invented at Georgia Tech, to utilize subsurface stressors to control strain and therefore material properties and growth kinetics in the material above the stressors...

Methodology to measure strains at high temperatures using electrical strain gages with free filaments

International Nuclear Information System (INIS)

Atanazio Filho, Nelson N.; Gomes, Paulo T. Vida; Scaldaferri, Denis H.B.; Silva, Luiz L. da; Rabello, Emerson G.; Mansur, Tanius R.

2013-01-01

An experimental methodology used for strains measuring at high temperatures is show in this work. In order to do the measurements, it was used electric strain gages with loose filaments attached to a stainless steel 304 beam with specific cements. The beam has triangular shape and a constant thickness, so the strain is the same along its length. Unless the beam surface be carefully prepared, the strain gage attachment is not efficient. The showed results are for temperatures ranging from 20 deg C to 300 deg C, but the experimental methodology could be used to measure strains at a temperature up to 900 deg C. Analytical calculations based on solid mechanics were used to verify the strain gage electrical installation and the measured strains. At a first moment, beam deformations as a temperature function were plotted. After that, beam deformations with different weighs were plotted as a temperature function. The results shown allowed concluding that the experimental methodology is trustable to measure strains at temperatures up to 300 deg C. (author)

Strain mapping near a triple junction in strained Ni-based alloy using EBSD and biaxial nanogauges

Energy Technology Data Exchange (ETDEWEB)

Clair, A. [Laboratoire Interdisciplinaire Carnot de Bourgogne, UMR 5209 CNRS, Universite de Bourgogne, 9 Avenue Alain Savary, BP 47870, 21078 Dijon Cedex (France); Foucault, M.; Calonne, O. [Areva ANP, Centre Technique Departement Corrosion-Chimie, 30 Bd de l' industrie, BP 181, 71205 Le Creusot (France); Lacroute, Y.; Markey, L.; Salazar, M.; Vignal, V. [Laboratoire Interdisciplinaire Carnot de Bourgogne, UMR 5209 CNRS, Universite de Bourgogne, 9 Avenue Alain Savary, BP 47870, 21078 Dijon Cedex (France); Finot, E., E-mail: Eric.Finot@u-bourgogne.fr [Laboratoire Interdisciplinaire Carnot de Bourgogne, UMR 5209 CNRS, Universite de Bourgogne, 9 Avenue Alain Savary, BP 47870, 21078 Dijon Cedex (France)

2011-05-15

Research highlights: > Surface strains measured using nanogauge were compared to the texture obtained by EBSD. > Statistics of the principal strain discern the grains according to the Schmid factor. > Strain hotspots were localized near a triple junction of alloy 600 under tensile loading. > Asymetrical profile of the GB strains is a criterion for surface cracking initiation. - Abstract: A key element for analyzing the crack initiation in strained polycrystalline alloys is the local quantification of the surface strain distribution according to the grain texture. Using electron backscattered diffraction, the local microstructure was determined to both localize a triple junction and deduce the local Schmid factors. Kernel average misorientation (KAM) was also used to map the areas of defect concentration. The maximum principal strain and the in-plane shear strain were quantified using the biaxial nanogauge. Distortions of the array of nanodots used as spot markers were analyzed near the triple junction. The crystallographic orientation and the surface strain were then investigated both statistically for each grain and locally at the grain boundaries. The superimposition of microstructure and strain maps allows the high strain gradient (reaching 3-fold the applied strain) to be localized at preferential grain boundaries near the triple junction. The Schmid factors and the KAM were compared to the maximum principal strain and the in-plane shear strain respectively. The polycrystalline deformation was attributable first to the rotation of some grains, followed by the elongation of all grains along their preferential activated slip systems.

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.

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

Directory of Open Access Journals (Sweden)

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.

Solitary waves in morphogenesis: Determination fronts as strain-cued strain transformations among automatous cells

Science.gov (United States)

Cox, Brian N.; Landis, Chad M.

2018-02-01

We present a simple theory of a strain pulse propagating as a solitary wave through a continuous two-dimensional population of cells. A critical strain is assumed to trigger a strain transformation, while, simultaneously, cells move as automata to tend to restore a preferred cell density. We consider systems in which the strain transformation is a shape change, a burst of proliferation, or the commencement of growth (which changes the shape of the population sheet), and demonstrate isomorphism among these cases. Numerical and analytical solutions describe a strain pulse whose height does not depend on how the strain disturbance was first launched, or the rate at which the strain transformation is achieved, or the rate constant in the rule for the restorative cell motion. The strain pulse is therefore very stable, surviving the imposition of strong perturbations: it would serve well as a timing signal in development. The automatous wave formulation is simple, with few model parameters. A strong case exists for the presence of a strain pulse during amelogenesis. Quantitative analysis reveals a simple relationship between the velocity of the leading edge of the pulse in amelogenesis and the known speed of migration of ameloblast cells. This result and energy arguments support the depiction of wave motion as an automatous cell response to strain, rather than as a response to an elastic energy gradient. The theory may also contribute to understanding the determination front in somitogenesis, moving fronts of convergent-extension transformation, and mitotic wavefronts in the syncytial drosophila embryo.

Mycobacterium tuberculosis strains exhibit differential and strain-specific molecular signatures in pulmonary epithelial cells.

Science.gov (United States)

Mvubu, Nontobeko Eunice; Pillay, Balakrishna; Gamieldien, Junaid; Bishai, William; Pillay, Manormoney

2016-12-01

Although pulmonary epithelial cells are integral to innate and adaptive immune responses during Mycobacterium tuberculosis infection, global transcriptomic changes in these cells remain largely unknown. Changes in gene expression induced in pulmonary epithelial cells infected with M. tuberculosis F15/LAM4/KZN, F11, F28, Beijing and Unique genotypes were investigated by RNA sequencing (RNA-Seq). The Illumina HiSeq 2000 platform generated 50 bp reads that were mapped to the human genome (Hg19) using Tophat (2.0.10). Differential gene expression induced by the different strains in infected relative to the uninfected cells was quantified and compared using Cufflinks (2.1.0) and MeV (4.0.9), respectively. Gene expression varied among the strains with the total number of genes as follows: F15/LAM4/KZN (1187), Beijing (1252), F11 (1639), F28 (870), Unique (886) and H37Rv (1179). A subset of 292 genes was commonly induced by all strains, where 52 genes were down-regulated while 240 genes were up-regulated. Differentially expressed genes were compared among the strains and the number of induced strain-specific gene signatures were as follows: F15/LAM4/KZN (138), Beijing (52), F11 (255), F28 (55), Unique (186) and H37Rv (125). Strain-specific molecular gene signatures associated with functional pathways were observed only for the Unique and H37Rv strains while certain biological functions may be associated with other strain signatures. This study demonstrated that strains of M. tuberculosis induce differential gene expression and strain-specific molecular signatures in pulmonary epithelial cells. Specific signatures induced by clinical strains of M. tuberculosis can be further explored for novel host-associated biomarkers and adjunctive immunotherapies. Copyright © 2016 Elsevier Ltd. All rights reserved.

Toxicity of bifenazate and its principal active metabolite, diazene, to Tetranychus urticae and Panonychus citri and their relative toxicity to the predaceous mites, Phytoseiulus persimilis and Neoseiulus californicus.

Science.gov (United States)

Ochiai, Noriaki; Mizuno, Masayuki; Mimori, Norihiko; Miyake, Toshihiko; Dekeyser, Mark; Canlas, Liza Jara; Takeda, Makio

2007-01-01

Bifenazate is a novel carbazate acaricide discovered by Uniroyal Chemical (now Chemtura Corporation) for the control of phytophagous mites infesting agricultural and ornamental crops. Its acaricidal activity and that of its principal active metabolite, diazene, were characterized. Bifenazate and diazene had high toxicity and specificity both orally and topically to all life stages of Tetranychus urticae and Panonychus citri. Acute poisoning was observed with no temperature dependency. No cross-resistance was found to mites resistant to several other classes of acaricides, such as tebufenpyrad, etoxazole, fenbutatin oxide and dicofol. Bifenazate remained effective for a long time with only about a 10% loss of efficacy on T. urticae after 1 month of application in the field. All stages of development of the predatory mites, Phytoseiulus persimilis and Neoseiulus californicus, survived treatment by both bifenazate and diazene. When adult females of the two predatory mite species were treated with either bifenazate or diazene, they showed a normal level of fecundity and predatory activity in the laboratory, effectively suppressing spider mite population growth. Even when the predators were fed spider mite eggs that had been treated previously with bifenazate, they survived. These findings indicate that bifenazate is a very useful acaricide giving high efficacy, long-lasting activity and excellent selectivity for spider mites. It is, therefore, concluded that bifenazate is an ideal compound for controlling these pest mites.

Flexible piezotronic strain sensor.

Science.gov (United States)

Zhou, Jun; Gu, Yudong; Fei, Peng; Mai, Wenjie; Gao, Yifan; Yang, Rusen; Bao, Gang; Wang, Zhong Lin

2008-09-01

Strain sensors based on individual ZnO piezoelectric fine-wires (PFWs; nanowires, microwires) have been fabricated by a simple, reliable, and cost-effective technique. The electromechanical sensor device consists of a single electrically connected PFW that is placed on the outer surface of a flexible polystyrene (PS) substrate and bonded at its two ends. The entire device is fully packaged by a polydimethylsiloxane (PDMS) thin layer. The PFW has Schottky contacts at its two ends but with distinctly different barrier heights. The I- V characteristic is highly sensitive to strain mainly due to the change in Schottky barrier height (SBH), which scales linear with strain. The change in SBH is suggested owing to the strain induced band structure change and piezoelectric effect. The experimental data can be well-described by the thermionic emission-diffusion model. A gauge factor of as high as 1250 has been demonstrated, which is 25% higher than the best gauge factor demonstrated for carbon nanotubes. The strain sensor developed here has applications in strain and stress measurements in cell biology, biomedical sciences, MEMS devices, structure monitoring, and more.

Molecular typing of Brucella melitensis endemic strains and differentiation from the vaccine strain Rev-1.

Science.gov (United States)

Noutsios, Georgios T; Papi, Rigini M; Ekateriniadou, Loukia V; Minas, Anastasios; Kyriakidis, Dimitrios A

2012-03-01

In the present study forty-four Greek endemic strains of Br. melitensis and three reference strains were genotyped by Multi locus Variable Number Tandem Repeat (ML-VNTR) analysis based on an eight-base pair tandem repeat sequence that was revealed in eight loci of Br. melitensis genome. The forty-four strains were discriminated from the vaccine strain Rev-1 by Restriction Fragment Length Polymorphism (RFLP) and Denaturant Gradient Gel Electrophoresis (DGGE). The ML-VNTR analysis revealed that endemic, reference and vaccine strains are genetically closely related, while most of the loci tested (1, 2, 4, 5 and 7) are highly polymorphic with Hunter-Gaston Genetic Diversity Index (HGDI) values in the range of 0.939 to 0.775. Analysis of ML-VNTRs loci stability through in vitro passages proved that loci 1 and 5 are non stable. Therefore, vaccine strain can be discriminated from endemic strains by allele's clusters of loci 2, 4, 6 and 7. RFLP and DGGE were also employed to analyse omp2 gene and reveled different patterns among Rev-1 and endemic strains. In RFLP, Rev-1 revealed three fragments (282, 238 and 44 bp), while endemic strains two fragments (238 and 44 bp). As for DGGE, the electrophoretic mobility of Rev-1 is different from the endemic strains due to heterologous binding of DNA chains of omp2a and omp2b gene. Overall, our data show clearly that it is feasible to genotype endemic strains of Br. melitensis and differentiate them from vaccine strain Rev-1 with ML-VNTR, RFLP and DGGE techniques. These tools can be used for conventional investigations in brucellosis outbreaks.

Effects of the Strain Rate Sensitivity and Strain Hardening on the Saturated Impulse of Plates

Directory of Open Access Journals (Sweden)

Ling Zhu

Full Text Available Abstract This paper studies the stiffening effects of the material strain rate sensitivity and strain hardening on the saturated impulse of elastic, perfectly plastic plates. Finite element (FE code ABAQUS is employed to simulate the elastoplastic response of square plates under rectangular pressure pulse. Rigid-plastic analyses for saturated impulse, which consider strain rate sensitivity and strain hardening, are conducted. Satisfactory agreement between the finite element models (FEM and predictions of the rigid-plastic analysis is obtained, which verifies that the proposed rigid-plastic methods are effective to solve the problem including strain rate sensitivity and strain hardening. The quantitative results for the scale effect of the strain rate sensitivity are given. The results for the stiffening effects suggest that two general stiffening factors n 1 and n 2, which characterizes the strain rate sensitivity and strain hardening effect, respectively can be defined. The saturated displacement is inversely proportional to the stiffening factors (i.e. n 1 and n 2 and saturated impulse is inversely proportional to the square roots of the stiffening factors (i.e. n 1 and n 2. Formulae for displacement and saturated impulse are proposed based on the empirical analysis.

Effect of strain rate and temperature at high strains on fatigue behavior of SAP alloys

DEFF Research Database (Denmark)

Blucher, J.T.; Knudsen, Per; Grant, N.J.

1968-01-01

Fatigue behavior of three SAP alloys of two nominal compositions (7 and 13% Al2O3) was studied in terms of strain rate and temperature at high strains; strain rate had no effect on life at 80 F, but had increasingly greater effect with increasing temperature above 500 F; life decreased with decre......Fatigue behavior of three SAP alloys of two nominal compositions (7 and 13% Al2O3) was studied in terms of strain rate and temperature at high strains; strain rate had no effect on life at 80 F, but had increasingly greater effect with increasing temperature above 500 F; life decreased...

Genome-Wide Transcription Study of Cryptococcus neoformans H99 Clinical Strain versus Environmental Strains.

Directory of Open Access Journals (Sweden)

Elaheh Movahed

Full Text Available The infection of Cryptococcus neoformans is acquired through the inhalation of desiccated yeast cells and basidiospores originated from the environment, particularly from bird's droppings and decaying wood. Three environmental strains of C. neoformans originated from bird droppings (H4, S48B and S68B and C. neoformans reference clinical strain (H99 were used for intranasal infection in C57BL/6 mice. We showed that the H99 strain demonstrated higher virulence compared to H4, S48B and S68B strains. To examine if gene expression contributed to the different degree of virulence among these strains, a genome-wide microarray study was performed to inspect the transcriptomic profiles of all four strains. Our results revealed that out of 7,419 genes (22,257 probes examined, 65 genes were significantly up-or down-regulated in H99 versus H4, S48B and S68B strains. The up-regulated genes in H99 strain include Hydroxymethylglutaryl-CoA synthase (MVA1, Mitochondrial matrix factor 1 (MMF1, Bud-site-selection protein 8 (BUD8, High affinity glucose transporter 3 (SNF3 and Rho GTPase-activating protein 2 (RGA2. Pathway annotation using DAVID bioinformatics resource showed that metal ion binding and sugar transmembrane transporter activity pathways were highly expressed in the H99 strain. We suggest that the genes and pathways identified may possibly play crucial roles in the fungal pathogenesis.

Life Stress, Strain, and Deviance Across Schools: Testing the Contextual Version of General Strain Theory in China.

Science.gov (United States)

Zhang, Jinwu; Liu, Jianhong; Wang, Xin; Zou, Anquan

2017-08-01

General Strain Theory delineates different types of strain and intervening processes from strain to deviance and crime. In addition to explaining individual strain-crime relationship, a contextualized version of general strain theory, which is called the Macro General Strain Theory, has been used to analyze how aggregate variables influence aggregate and individual deviance and crime. Using a sample of 1,852 students (Level 1) nested in 52 schools (Level 2), the current study tests the Macro General Strain Theory using Chinese data. The results revealed that aggregate life stress and strain have influences on aggregate and individual deviance, and reinforce the individual stress-deviance association. The current study contributes by providing the first Macro General Strain Theory test based on Chinese data and offering empirical evidence for the multilevel intervening processes from strain to deviance. Limitations and future research directions are discussed.

Strain Pattern in Supercooled Liquids

Science.gov (United States)

Illing, Bernd; Fritschi, Sebastian; Hajnal, David; Klix, Christian; Keim, Peter; Fuchs, Matthias

2016-11-01

Investigations of strain correlations at the glass transition reveal unexpected phenomena. The shear strain fluctuations show an Eshelby-strain pattern [˜cos (4 θ ) /r2 ], characteristic of elastic response, even in liquids, at long times. We address this using a mode-coupling theory for the strain fluctuations in supercooled liquids and data from both video microscopy of a two-dimensional colloidal glass former and simulations of Brownian hard disks. We show that the long-ranged and long-lived strain signatures follow a scaling law valid close to the glass transition. For large enough viscosities, the Eshelby-strain pattern is visible even on time scales longer than the structural relaxation time τ and after the shear modulus has relaxed to zero.

Biological assay of attenuated strain NADL-2 and virulent strain NADL-8 of porcine parvovirus.

Science.gov (United States)

Mengeling, W L; Pejsak, Z; Paul, P S

1984-11-01

Attenuated strain NADL-2 and virulent strain NADL-8 of porcine parvovirus (PPV) were titrated in vivo and in vitro under similar conditions to provide a better understanding of some of the factors involved in virulence of PPV in causing maternal reproductive failure of swine. Both strains cause fetal death when they are injected directly into fetal fluids, but only strain NADL-8 does so when administered to pregnant swine. The strains were tested for their hemagglutinating activity (HA), median cell culture infective dose (CCID50), median fetal infective dose (FID50), and median fetal lethal dose (FLD50). The FID50 and FLD50 were determined by injecting virus directly into the amniotic fluid of fetuses in utero at 44 +/- 2 days of gestation and collecting the fetuses 15 +/- 1 days later. Both strains had an HA titer of 64, suggesting that there is a similar number of virions in stock preparations. However, other measurements differed markedly. The CCID50, FID50, and FLD50 were 10(5.5), 10(3.5), and 10(0.5), respectively, for strain NADL-2, and 10(4.5), 10(7.7), and 10(6.3), respectively, for strain NADL-8. Collectively, the values indicate that more than 10,000 times as much strain NADL-2 would need to reach the conceptus transplacentally to establish infection. These observations may help to explain the different consequences of oronasal exposure of pregnant swine to these strains of PPV.

Haldane model under nonuniform strain

Science.gov (United States)

Ho, Yen-Hung; Castro, Eduardo V.; Cazalilla, Miguel A.

2017-10-01

We study the Haldane model under strain using a tight-binding approach, and compare the obtained results with the continuum-limit approximation. As in graphene, nonuniform strain leads to a time-reversal preserving pseudomagnetic field that induces (pseudo-)Landau levels. Unlike a real magnetic field, strain lifts the degeneracy of the zeroth pseudo-Landau levels at different valleys. Moreover, for the zigzag edge under uniaxial strain, strain removes the degeneracy within the pseudo-Landau levels by inducing a tilt in their energy dispersion. The latter arises from next-to-leading order corrections to the continuum-limit Hamiltonian, which are absent for a real magnetic field. We show that, for the lowest pseudo-Landau levels in the Haldane model, the dominant contribution to the tilt is different from graphene. In addition, although strain does not strongly modify the dispersion of the edge states, their interplay with the pseudo-Landau levels is different for the armchair and zigzag ribbons. Finally, we study the effect of strain in the band structure of the Haldane model at the critical point of the topological transition, thus shedding light on the interplay between nontrivial topology and strain in quantum anomalous Hall systems.

Strain preservation of experimental animals: vitrification of two-cell stage embryos for multiple mouse strains.

Science.gov (United States)

Eto, Tomoo; Takahashi, Riichi; Kamisako, Tsutomu

2015-04-01

Strain preservation of experimental animals is crucial for experimental reproducibility. Maintaining complete animal strains, however, is costly and there is a risk for genetic mutations as well as complete loss due to disasters or illness. Therefore, the development of effective vitrification techniques for cryopreservation of multiple experimental animal strains is important. We examined whether a vitrification method using cryoprotectant solutions, P10 and PEPeS, is suitable for preservation of multiple inbred and outbred mouse strains. First, we investigated whether our vitrification method using cryoprotectant solutions was suitable for two-cell stage mouse embryos. In vitro development of embryos exposed to the cryoprotectant solutions was similar to that of fresh controls. Further, the survival rate of the vitrified embryos was extremely high (98.1%). Next, we collected and vitrified two-cell stage embryos of 14 mouse strains. The average number of embryos obtained from one female was 7.3-33.3. The survival rate of vitrified embryos ranged from 92.8% to 99.1%, with no significant differences among mouse strains. In vivo development did not differ significantly between fresh controls and vitrified embryos of each strain. For strain preservation using cryopreserved embryos, two offspring for inbred lines and one offspring for outbred lines must be produced from two-cell stage embryos collected from one female. The expected number of surviving fetuses obtained from embryos collected from one female of either the inbred or outbred strains ranged from 2.9 to 19.5. The findings of the present study indicated that this vitrification method is suitable for strain preservation of multiple mouse strains. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

Colony Dimorphism in Bradyrhizobium Strains

Science.gov (United States)

Sylvester-Bradley, Rosemary; Thornton, Philip; Jones, Peter

1988-01-01

Ten isolates of Bradyrhizobium spp. which form two colony types were studied; the isolates originated from a range of legume species. The two colony types differed in the amount of gum formed or size or both, depending on the strain. Whole 7-day-old colonies of each type were subcultured to determine the proportion of cells which had changed to the other type. An iterative computerized procedure was used to determine the rate of switching per generation between the two types and to predict proportions reached at equilibrium for each strain. The predicted proportions of the wetter (more gummy) or larger colony type at equilibrium differed significantly between strains, ranging from 0.9999 (strain CIAT 2383) to 0.0216 (strain CIAT 2469), because some strains switched faster from dry to wet (or small to large) and others switched faster from wet to dry (or large to small). Predicted equilibrium was reached after about 140 generations in strain USDA 76. In all but one strain (CIAT 3030) the growth rate of the wetter colony type was greater than or similar to that of the drier type. The mean difference in generation time between the two colony types was 0.37 h. Doubling times calculated for either colony type after 7 days of growth on the agar surface ranged from 6.0 to 7.3 h. The formation of two persistent colony types by one strain (clonal or colony dimorphism) may be a common phenomenon among Bradyrhizobium strains. Images PMID:16347599

Conduction band structure and electron mobility in uniaxially strained Si via externally applied strain in nanomembranes

Energy Technology Data Exchange (ETDEWEB)

Chen Feng [Xi' an Jiaotong University, Xi' an, Shaanxi 710049 (China); Euaruksakul, Chanan; Himpsel, F J; Lagally, Max G [University of Wisconsin-Madison, Madison, WI 53706 (United States); Liu Zheng; Liu Feng, E-mail: lagally@engr.wisc.edu [University of Utah, Salt Lake City, UT 84112 (United States)

2011-08-17

Strain changes the band structure of semiconductors. We use x-ray absorption spectroscopy to study the change in the density of conduction band (CB) states when silicon is uniaxially strained along the [1 0 0] and [1 1 0] directions. High stress can be applied to silicon nanomembranes, because their thinness allows high levels of strain without fracture. Strain-induced changes in both the sixfold degenerate {Delta} valleys and the eightfold degenerate L valleys are determined quantitatively. The uniaxial deformation potentials of both {Delta} and L valleys are directly extracted using a strain tensor appropriate to the boundary conditions, i.e., confinement in the plane in the direction orthogonal to the straining direction, which correspond to those of strained CMOS in commercial applications. The experimentally determined deformation potentials match the theoretical predictions well. We predict electron mobility enhancement created by strain-induced CB modifications.

Mechanism of action and selective toxicity of ascamycin, a nucleoside antibiotic.

OpenAIRE

Osada, H; Isono, K

1985-01-01

An unidentified Streptomyces sp. produces two nucleoside antibiotics, ascamycin and its dealanyl derivative. In contrast to the broad antibacterial activity of dealanylascamycin against various gram-negative and gram-positive bacteria, ascamycin showed selective toxicity against Xanthomonas citri and X. oryzae. Both ascamycin and dealanylascamycin inhibited the protein synthesis of X. citri, but only dealanylascamycin inhibited that of Escherichia coli. In cell-free systems from E. coli and X...

Effect of whole yeast cell product supplementation (CitriStim®) on immune responses and cecal microflora species in pullet and layer chickens during an experimental coccidial challenge.

Science.gov (United States)

Markazi, Ashley D; Perez, Victor; Sifri, Mamduh; Shanmugasundaram, Revathi; Selvaraj, Ramesh K

2017-07-01

Three separate experiments were conducted to study the effects of whole yeast cell product supplementation in pullets and layer hens. Body weight gain, fecal and intestinal coccidial oocyst counts, cecal microflora species, cytokine mRNA amounts, and CD4+ and CD8+ T-cell populations in the cecal tonsils were analyzed following an experimental coccidial infection. In Experiment I, day-old Leghorn layer chicks were fed 3 experimental diets with 0, 0.1, or 0.2% whole yeast cell product (CitriStim®, ADM, Decatur, IL). At 21 d of age, birds were challenged with 1 × 105 live coccidial oocysts. Supplementation with whole yeast cell product decreased the fecal coccidial oocyst count at 7 (P = 0.05) and 8 (P product and challenged with 1 × 105 live coccidial oocysts on d 25 of whole yeast cell product feeding. Supplementation with whole yeast cell product decreased the coccidial oocyst count in the intestinal content (P product increased relative proportion of Lactobacillus (P product decreased CD8+ T cell percentages (P product and challenged with 1 × 105 live coccidial oocysts on d 66 of whole yeast cell product feeding. At 5 d post-coccidial challenge, whole yeast cell product supplementation down-regulated (P = 0.01) IL-10 mRNA amount. It could be concluded that supplementing whole yeast cell product can help minimize coccidial infection in both growing pullets and layer chickens. © 2017 Poultry Science Association Inc.

Studies on Drosophila radiosensitive strains

International Nuclear Information System (INIS)

Varentsova, E.P.; Zakharov, I.A.

1976-01-01

45 of radiosensitive strains of Drosophila melanogaster were isolated by Curly/Lobe technique after EMS treatment of Livadia population males. The lethality of non-Curly late larvae after gamma-irradiation (4000r) characterized radiosensitivity strains. Most of them exhibited higher frequency of the spontaneous dominant lethals (up to 69%). The males of 6 strains were semi-sterile. 5 of these strains exhibited higher frequency of X-chromosome non-disjunction

Segregation of genes from donor strain during the production of recombinant congenic strains.

Science.gov (United States)

van Zutphen, L F; Den Bieman, M; Lankhorst, A; Demant, P

1991-07-01

Recombinant congenic strains (RCS) constitute a set of inbred strains which are designed to dissect the genetic control of multigenic traits, such as tumour susceptibility or disease resistance. Each RCS contains a small fraction of the genome of a common donor strain, while the majority of genes stem from a common background strain. We tested at two stages of the inbreeding process in 20 RCS, derived from BALB/cHeA and STS/A, to see whether alleles from the STS/A donor strain are distributed over the RCS in a ratio as would theoretically be expected. Four marker genes (Pep-3; Pgm-1; Gpi-1 and Es-3) located at 4 different chromosomes were selected and the allelic distribution was tested after 3-4 and after 12 generations of inbreeding. The data obtained do not significantly deviate from the expected pattern, thus supporting the validity of the concept of RCS.

A new strain gage method for measuring the contractile strain ratio of Zircaloy tubing

International Nuclear Information System (INIS)

Hwang, S.K.; Sabol, G.P.

1988-01-01

An improved strain gage method for determining the contractile strain ratio (CSR) of Zircaloy tubing was developed. The new method consists of a number of load-unload cyclings at approximately 0.2% plastic strain interval. With this method the CSR of Zircaloy-4 tubing could be determined accurately because it was possible to separate the plastic strains from the elastic strain involvement. The CSR values determined by use of the new method were in good agreement with those calculated from conventional post-test manual measurements. The CSR of the tubing was found to decrease with the amount of deformation during testing because of uneven plastic flow in the gage section. A new technique of inscribing gage marks by use of a YAG laser is discussed. (orig.)

Strain localization band width evolution by electronic speckle pattern interferometry strain rate measurement

Energy Technology Data Exchange (ETDEWEB)

Guelorget, Bruno [Institut Charles Delaunay-LASMIS, Universite de technologie de Troyes, FRE CNRS 2848, 12 rue Marie Curie, B.P. 2060, 10010 Troyes Cedex (France)], E-mail: bruno.guelorget@utt.fr; Francois, Manuel; Montay, Guillaume [Institut Charles Delaunay-LASMIS, Universite de technologie de Troyes, FRE CNRS 2848, 12 rue Marie Curie, B.P. 2060, 10010 Troyes Cedex (France)

2009-04-15

In this paper, electronic speckle pattern interferometry strain rate measurements are used to quantify the width of the strain localization band, which occurs when a sheet specimen is submitted to tension. It is shown that the width of this band decreases with increasing strain. Just before fracture, this measured width is about five times wider than the shear band and the initial sheet thickness.

Thermal strain measurement of EAST W/Cu divertor structure using electric resistance strain gauges

Energy Technology Data Exchange (ETDEWEB)

Wang, Xingli [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, 230031 (China); Science Island Branch of Graduate School, University of Science & Technology of China, Hefei, 230031 (China); Wang, Wanjing, E-mail: wjwang@ipp.ac.cn [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, 230031 (China); Wang, Jichao [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, 230031 (China); Wei, Ran; Sun, Zhaoxuan [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, 230031 (China); Science Island Branch of Graduate School, University of Science & Technology of China, Hefei, 230031 (China); Li, Qiang; Xie, Chunyi [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, 230031 (China); Chen, Hong-En; Wang, Kaiqiang; Wu, Lei; Chen, Zhenmao [State Key Lab for Strength and Vibration of Mechanical Structures, Xi’an Jiaotong University (China); Luo, Guang-Nan [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, 230031 (China); Science Island Branch of Graduate School, University of Science & Technology of China, Hefei, 230031 (China); Hefei Center for Physical Science and Technology, Hefei, 230022 (China); Hefei Science Center of Chinese Academy of Sciences, Hefei, 230027 (China)

2016-12-15

Highlights: • To understand the service behavior of W/Cu divertor, an electrical resistance strain gauge system had been introduced in a thermal strain measurement experiment. • The measurement system successfully finished the experiment and obtained valued thermal strain data. • Two thermomechanical analyses had also been carried out and compared with the measurement results. • Experiment results corresponded well to simulations and threw a light upon the failure of W/Cu divertor in the previous baking tests. - Abstract: W/Cu divertor has complex structure and faces extreme work environment in EAST Tokamak device. To measure its thermal strain shall be a valued way to understand its service behavior and then optimize its design and manufacturing process. This work presents a preliminary study on measuring thermal strain of EAST W/Cu divertor structure using electric resistance strain gauges. Eight gauges had been used in the experiment and the heating temperature had been set to 230 °C with respect to the work temperature. To realize the measuring experiment, an appropriate fixing method of gauges in divertor narrow spaces had been taken and tested, which could not only withstand high temperature but also had no damage to the divertor sample. The measurement results were that three gauges showed positive strain while other three showed negative strain after having been compensated, which corresponded to tensile stress and compressed stress respectively. Two thermomechanical simulations had also been carried out and used for comparing with the experiment.

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

Science.gov (United States)

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

2015-01-01

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

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

Directory of Open Access Journals (Sweden)

Xavier Martini

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

Rapid establishment of a regular distribution of adult tropical Drosophila parasitoids in a multi-patch environment by patch defence behaviour.

Science.gov (United States)

de Jong, Peter W; Hemerik, Lia; Gort, Gerrit; van Alphen, Jacques J M

2011-01-01

Females of the larval parasitoid of Drosophila, Asobara citri, from sub-Saharan Africa, defend patches with hosts by fighting and chasing conspecific females upon encounter. Females of the closely related, palearctic species Asobara tabida do not defend patches and often search simultaneously in the same patch. The effect of patch defence by A. citri females on their distribution in a multi-patch environment was investigated, and their distributions were compared with those of A. tabida. For both species 20 females were released from two release-points in replicate experiments. Females of A. citri quickly reached a regular distribution across 16 patches, with a small variance/mean ratio per patch. Conversely, A. tabida females initially showed a clumped distribution, and after gradual dispersion, a more Poisson-like distribution across patches resulted (variance/mean ratio was closer to 1 and higher than for A. citri). The dispersion of A. tabida was most probably an effect of exploitation: these parasitoids increasingly made shorter visits to already exploited patches. We briefly discuss hypotheses on the adaptive significance of patch defence behaviour or its absence in the light of differences in the natural history of both parasitoid species, notably the spatial distribution of their hosts.

Noninvasive characterization of carotid plaque strain.

Science.gov (United States)

Khan, Amir A; Sikdar, Siddhartha; Hatsukami, Thomas; Cebral, Juan; Jones, Michael; Huston, John; Howard, George; Lal, Brajesh K

2017-06-01

Current risk stratification of internal carotid artery plaques based on diameter-reducing percentage stenosis may be unreliable because ischemic stroke results from plaque disruption with atheroembolization. Biomechanical forces acting on the plaque may render it vulnerable to rupture. The feasibility of ultrasound-based quantification of plaque displacement and strain induced by hemodynamic forces and their relationship to high-risk plaques have not been determined. We studied the feasibility and reliability of carotid plaque strain measurement from clinical B-mode ultrasound images and the relationship of strain to high-risk plaque morphology. We analyzed carotid ultrasound B-mode cine loops obtained in patients with asymptomatic ≥50% stenosis during routine clinical scanning. Optical flow methods were used to quantify plaque motion and shear strain during the cardiac cycle. The magnitude (maximum absolute shear strain rate [MASSR]) and variability (entropy of shear strain rate [ESSR] and variance of shear strain rate [VSSR]) of strain were combined into a composite shear strain index (SSI), which was assessed for interscan repeatability and correlated with plaque echolucency. Nineteen patients (mean age, 70 years) constituting 36 plaques underwent imaging; 37% of patients (n = 7) showed high strain (SSI ≥0.5; MASSR, 2.2; ESSR, 39.7; VSSR, 0.03) in their plaques; the remaining clustered into a low-strain group (SSI routine B-mode imaging using clinical ultrasound machines. High plaque strain correlates with known high-risk echolucent morphology. Strain measurement can complement identification of patients at high risk for plaque disruption and stroke. Copyright © 2017 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.

Revealing differences in metabolic flux distributions between a mutant strain and its parent strain Gluconacetobacter xylinus CGMCC 2955.

Directory of Open Access Journals (Sweden)

Cheng Zhong

Full Text Available A better understanding of metabolic fluxes is important for manipulating microbial metabolism toward desired end products, or away from undesirable by-products. A mutant strain, Gluconacetobacter xylinus AX2-16, was obtained by combined chemical mutation of the parent strain (G. xylinus CGMCC 2955 using DEC (diethyl sulfate and LiCl. The highest bacterial cellulose production for this mutant was obtained at about 11.75 g/L, which was an increase of 62% compared with that by the parent strain. In contrast, gluconic acid (the main byproduct concentration was only 5.71 g/L for mutant strain, which was 55.7% lower than that of parent strain. Metabolic flux analysis indicated that 40.1% of the carbon source was transformed to bacterial cellulose in mutant strain, compared with 24.2% for parent strain. Only 32.7% and 4.0% of the carbon source were converted into gluconic acid and acetic acid in mutant strain, compared with 58.5% and 9.5% of that in parent strain. In addition, a higher flux of tricarboxylic acid (TCA cycle was obtained in mutant strain (57.0% compared with parent strain (17.0%. It was also indicated from the flux analysis that more ATP was produced in mutant strain from pentose phosphate pathway (PPP and TCA cycle. The enzymatic activity of succinate dehydrogenase (SDH, which is one of the key enzymes in TCA cycle, was 1.65-fold higher in mutant strain than that in parent strain at the end of culture. It was further validated by the measurement of ATPase that 3.53-6.41 fold higher enzymatic activity was obtained from mutant strain compared with parent strain.

Mumps Hoshino and Torii vaccine strains were distinguished from circulating wild strains.

Science.gov (United States)

Sawada, Akihito; Yamaji, Yoshiaki; Nakayama, Tetsuo

2013-06-01

Aseptic meningitis and acute parotitis have been observed after mumps vaccination. Mumps outbreaks have been reported in Japan because of low vaccine coverage, and molecular differentiation is required to determine whether these cases are vaccine associated. RT-nested PCR was performed in the small hydrophobic gene region, and viruses were differentiated by restriction fragment length polymorphism assay. A total of 584 nucleotides were amplified. The PCR product of the Hoshino strain was cut into two fragments (313 and 271 nucleotides) by MfeI; that of the Torii strain was digested with EcoT22I, resulting in 332- and 252-nucleotide fragments. Both strains were genotype B and had an XbaI site, resulting in two fragments: 299 and 285 nucleotides. Current circulating wild types were cut only by XbaI or MfeI. However, the MfeI site of the wild types was different from that of the Hoshino strain, resulting in 451- and 133-nucleotide fragments. Using three restriction enzymes, two mumps vaccine strains were distinguished from wild types, and this separation was applied to the identification of vaccine-related adverse events.

An Embeddable Strain Sensor with 30 Nano-Strain Resolution Based on Optical Interferometry

Directory of Open Access Journals (Sweden)

Chen Zhu

2018-04-01

Full Text Available A cost-effective, robust and embeddable optical interferometric strain sensor with nanoscale strain resolution is presented in this paper. The sensor consists of an optical fiber, a quartz rod with one end coated with a thin gold layer, and two metal shells employed to transfer the strain and orient and protect the optical fiber and the quartz rod. The optical fiber endface, combining with the gold-coated surface, forms an extrinsic Fabry–Perot interferometer. The sensor was firstly calibrated, and the result showed that our prototype sensor could provide a measurement resolution of 30 nano-strain (nε and a sensitivity of 10.01 µε/µm over a range of 1000 µε. After calibration of the sensor, the shrinkage strain of a cubic brick of mortar in real time during the drying process was monitored. The strain sensor was compared with a commercial linear variable displacement transducer, and the comparison results in four weeks demonstrated that our sensor had much higher measurement resolution and gained more detailed and useful information. Due to the advantages of the extremely simple, robust and cost-effective configuration, it is believed that the sensor is significantly beneficial to practical applications, especially for structural health monitoring.

An ultrasensitive strain sensor with a wide strain range based on graphene armour scales.

Science.gov (United States)

Yang, Yi-Fan; Tao, Lu-Qi; Pang, Yu; Tian, He; Ju, Zhen-Yi; Wu, Xiao-Ming; Yang, Yi; Ren, Tian-Ling

2018-06-12

An ultrasensitive strain sensor with a wide strain range based on graphene armour scales is demonstrated in this paper. The sensor shows an ultra-high gauge factor (GF, up to 1054) and a wide strain range (ε = 26%), both of which present an advantage compared to most other flexible sensors. Moreover, the sensor is developed by a simple fabrication process. Due to the excellent performance, this strain sensor can meet the demands of subtle, large and complex human motion monitoring, which indicates its tremendous application potential in health monitoring, mechanical control, real-time motion monitoring and so on.

Strain rate measurement by Electronic Speckle Pattern Interferometry: A new look at the strain localization onset

International Nuclear Information System (INIS)

Guelorget, Bruno; Francois, Manuel; Vial-Edwards, Cristian; Montay, Guillaume; Daniel, Laurent; Lu, Jian

2006-01-01

In-plane Electronic Speckle Pattern Interferometry has been successfully used during tensile testing of semi-hard copper sheets in order to measure the strain rate. On one hand, heterogeneity in strain rate field has been found before the maximum of the tensile force (ε t ≅ 19.4 and 25.4%, respectively). Thus, a localization phenomenon occurs before the classic Considere's criterion (dF = 0) for the diffuse neck initiation. On the other hand, strain rate measurement before fracture shows the moment where one of the two slip band systems becomes predominant, then strain concentrates in a small area, the shear band. Uncertainty evaluation has been carried out, which shows a very good accuracy of the total strain and the strain rate measurements

Strain rate measurement by Electronic Speckle Pattern Interferometry: A new look at the strain localization onset

Energy Technology Data Exchange (ETDEWEB)

Guelorget, Bruno [Universite de Technologie de Troyes (UTT), Laboratoire des Systemes Mecaniques et d' ingenierie Simultanee (LASMIS, CNRS FRE 2719), 12 rue Marie Curie, B.P. 2060, 10010 Troyes Cedex (France)]. E-mail: bruno.guelorget@utt.fr; Francois, Manuel [Universite de Technologie de Troyes (UTT), Laboratoire des Systemes Mecaniques et d' ingenierie Simultanee (LASMIS, CNRS FRE 2719), 12 rue Marie Curie, B.P. 2060, 10010 Troyes Cedex (France); Vial-Edwards, Cristian [Departemento de Ingenieria Mecanica y Metalurgica, Pontificia Universidad Catolica de Chile, Vicuna Mackenna 4860, 6904411 Santiago (Chile); Montay, Guillaume [Universite de Technologie de Troyes (UTT), Laboratoire des Systemes Mecaniques et d' ingenierie Simultanee (LASMIS, CNRS FRE 2719), 12 rue Marie Curie, B.P. 2060, 10010 Troyes Cedex (France); Daniel, Laurent [Universite de Technologie de Troyes (UTT), Laboratoire des Systemes Mecaniques et d' ingenierie Simultanee (LASMIS, CNRS FRE 2719), 12 rue Marie Curie, B.P. 2060, 10010 Troyes Cedex (France); Lu, Jian [Universite de Technologie de Troyes (UTT), Laboratoire des Systemes Mecaniques et d' ingenierie Simultanee (LASMIS, CNRS FRE 2719), 12 rue Marie Curie, B.P. 2060, 10010 Troyes Cedex (France)

2006-01-15

In-plane Electronic Speckle Pattern Interferometry has been successfully used during tensile testing of semi-hard copper sheets in order to measure the strain rate. On one hand, heterogeneity in strain rate field has been found before the maximum of the tensile force ({epsilon} {sup t} {approx_equal} 19.4 and 25.4%, respectively). Thus, a localization phenomenon occurs before the classic Considere's criterion (dF = 0) for the diffuse neck initiation. On the other hand, strain rate measurement before fracture shows the moment where one of the two slip band systems becomes predominant, then strain concentrates in a small area, the shear band. Uncertainty evaluation has been carried out, which shows a very good accuracy of the total strain and the strain rate measurements.

Mechanical strength model for plastic bonded granular materials at high strain rates and large strains

International Nuclear Information System (INIS)

Browning, R.V.; Scammon, R.J.

1998-01-01

Modeling impact events on systems containing plastic bonded explosive materials requires accurate models for stress evolution at high strain rates out to large strains. For example, in the Steven test geometry reactions occur after strains of 0.5 or more are reached for PBX-9501. The morphology of this class of materials and properties of the constituents are briefly described. We then review the viscoelastic behavior observed at small strains for this class of material, and evaluate large strain models used for granular materials such as cap models. Dilatation under shearing deformations of the PBX is experimentally observed and is one of the key features modeled in cap style plasticity theories, together with bulk plastic flow at high pressures. We propose a model that combines viscoelastic behavior at small strains but adds intergranular stresses at larger strains. A procedure using numerical simulations and comparisons with results from flyer plate tests and low rate uniaxial stress tests is used to develop a rough set of constants for PBX-9501. Comparisons with the high rate flyer plate tests demonstrate that the observed characteristic behavior is captured by this viscoelastic based model. copyright 1998 American Institute of Physics

A strain gauge

DEFF Research Database (Denmark)

2017-01-01

The invention relates to a strain gauge of a carrier layer and a meandering measurement grid (101) positioned on the carrier layer, wherein the measurement grid comprises a number of measurement grid sections placed side by side with gaps in between, and a number of end loops (106) interconnecting...... relates to a method for manufacturing a strain gauge as mentioned above....

Origin of the Strain Sensitivity for an Organic Heptazole Thin-Film and Its Strain Gauge Application

Science.gov (United States)

Bae, Heesun; Jeon, Pyo Jin; Park, Ji Hoon; Lee, Kimoon

2018-04-01

The authors report on the origin of the strain sensitivity for an organic C26H16N2 (heptazole) thinfilm and its application for the detection of tensile strain. From the electrical characterization on the thin-film transistor adopting a heptazole channel, heptazole film exhibits p-channel conduction with a relatively low value of field-effect mobility (0.05 cm2/Vs), suggesting a hopping conduction behavior via hole carriers. By analyzing the strain and temperature dependences of the electrical conductivity, we reveal that the electrical conduction for a heptazole thin-film is dominated by the variable range hopping process with quite a large energy separation (224.9 meV) between the localized states under a relatively long attenuation length (10.46 Å). This indicates that a change in the inter-grain spacing that is much larger than the attenuation length is responsible for the reversible modification of electrical conductivity depending on strain for the heptazole film. By utilizing our heptazole thin-film both as a strain sensitive passive resistor and an active semiconducting channel layer, we can achieve a strain gauge device exhibiting reversible endurance for tensile strains up to 2.12%. Consequently, this study advances the understanding of the fundamental strain sensing mechanism in a heptazole thin-film toward finding a promise material with a strain gauge for applications as potential flexible devices and/or wearable electronics.

Asymptomatic bacteriuria Escherichia coli strains

DEFF Research Database (Denmark)

Hancock, Viktoria; Nielsen, E.M.; Klemm, Per

2006-01-01

Urinary tract infections (UTIs) affect millions of people each year. Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU) in humans. Persons affected by ABU may carry a particular E. coli strain for extended periods of time without any symptoms. In contrast...... to uropathogenic E. coli (UPEC) that cause symptomatic UTI, very little is known about the mechanisms by which these strains colonize the urinary tract. Here, we have investigated the growth characteristics in human urine as well as adhesin repertoire of nine ABU strains; the ability of ABU strains to compete...

Strain-controlled nonvolatile magnetization switching

Science.gov (United States)

Geprägs, S.; Brandlmaier, A.; Brandt, M. S.; Gross, R.; Goennenwein, S. T. B.

2014-11-01

We investigate different approaches towards a nonvolatile switching of the remanent magnetization in single-crystalline ferromagnets at room temperature via elastic strain using ferromagnetic thin film/piezoelectric actuator hybrids. The piezoelectric actuator induces a voltage-controllable strain along different crystalline directions of the ferromagnetic thin film, resulting in modifications of its magnetization by converse magnetoelastic effects. We quantify the magnetization changes in the hybrids via ferromagnetic resonance spectroscopy and superconducting quantum interference device magnetometry. These measurements demonstrate a significant strain-induced change of the magnetization, limited by an inefficient strain transfer and domain formation in the particular system studied. To overcome these obstacles, we address practicable engineering concepts and use a model to demonstrate that a strain-controlled, nonvolatile magnetization switching should be possible in appropriately engineered ferromagnetic/piezoelectric actuator hybrids.

Strain and strain rate by two-dimensional speckle tracking echocardiography in a maned wolf Strain e strain rate por meio de ecocardiogratia speckle traking bidimensional em um lobo-guará

Directory of Open Access Journals (Sweden)

Matheus M. Mantovani

2012-12-01

Full Text Available The measurement of cardiovascular features of wild animals is important, as is the measurement in pets, for the assessment of myocardial function and the early detection of cardiac abnormalities, which could progress to heart failure. Speckle tracking echocardiography (2D STE is a new tool that has been used in veterinary medicine, which demonstrates several advantages, such as angle independence and the possibility to provide the early diagnosis of myocardial alterations. The aim of this study was to evaluate the left myocardial function in a maned wolf by 2D STE. Thus, the longitudinal, circumferential and radial strain and strain rate were obtained, as well as, the radial and longitudinal velocity and displacement values, from the right parasternal long axis four-chamber view, the left parasternal apical four chamber view and the parasternal short axis at the level of the papillary muscles. The results of the longitudinal variables were -13.52±7.88, -1.60±1.05, 4.34±2.52 and 3.86±3.04 for strain (%, strain rate (1/s, displacement (mm and velocity (cm/s, respectively. In addition, the radial and circumferential Strain and Strain rate were 24.39±14.23, 1.86±0.95 and -13.69±6.53, -1.01±0.48, respectively. Thus, the present study provides the first data regarding the use of this tool in maned wolves, allowing a more complete quantification of myocardial function in this species.A obtenção de parâmetros cardiovasculares em animais selvagens são importantes de serem avaliados, assim como em animais de companhia, para a obtenção da função miocárdica e determinação precoce de alterações cardíacas que poderiam evoluir para insuficiência cardíaca . A ecocardiografia speckle tracking (2D STE é uma ferramenta nova que tem sido utilizada em medicina veterinária, a qual tem demonstrado várias vantagens quanto ao seu uso, como a independência do ângulo de insonação e a possibilidade de se obter o diagnóstico precoce de altera

Whitefly Pest Species (Homoptera: Aleyrodidae) on Citrus Trees

OpenAIRE

Katja Žanić; Sonja Kačić; Miro Katalinić

2000-01-01

Today, the Citrus whitefly, Dialeurodes citri (Ashmead), is a very important pest on all Citrus species throughout the citrus growing areas in Croatia. It causes direct damage by sucking the plant juice from the leaves. Furthermore, immatures excrete honeydew that stimulates sooy mold. The presence of sooty mold on contaminated leaves interferes with the photosynthesis of plants. Citrus fruits coated by sooty mold lose its market value. Because Dialeurodes citri is poorly known in Croatia, th...

Failure strains and proposed limit strains for an reactor pressure vessel under severe accident conditions

International Nuclear Information System (INIS)

Krieg, R.

2005-01-01

The local failure strains of essential design elements of a reactor vessel are investigated. The size influence of the structure is of special interest. Typical severe accident conditions including elevated temperatures and dynamic loads are considered. The main part of work consists of test families with specimens under uniaxial and biaxial load. Within one test family the specimen geometry and the load conditions are similar, but the size is varied up to reactor dimensions. Special attention is given to geometries with a hole or a notch causing non-uniform stress and strain distributions typical for the reactor vessel. A key problem is to determine the local failure strain. Here suitable methods had to be developed including the so-called 'vanishing gap method', and the 'forging die method'. They are based on post-test geometrical measurements of the fracture surfaces and reconstructions of the related strain fields using finite element models. The results indicate that stresses versus dimensionless deformations are approximately size independent up to failure for specimens of similar geometry under similar load conditions. Local failure strains could be determined. The values are rather high and size dependent. Statistical evaluation allow the proposal of limit strains which are also size dependent. If these limit strains are not exceeded, the structures will not fracture

General Strain Theory and Delinquency: Focusing on the Influences of Key Strain Characteristics on Delinquency

Science.gov (United States)

Moon, Byongook; Blurton, David; McCluskey, John D.

2008-01-01

The study examines the effects of recent, older, and chronic strains and of perceived injustice of strain on delinquency, sampling 777 Korean youth. Seven key strains most likely leading to delinquency, some of which were often overlooked in previous research, were included, and these are family conflict, parental punishment, teachers' punishment,…

Enzyme markers in inbred rat strains: genetics of new markers and strain profiles.

Science.gov (United States)

Adams, M; Baverstock, P R; Watts, C H; Gutman, G A

1984-08-01

Twenty-six inbred strains of the laboratory rat (Rattus norvegicus) were examined for electrophoretic variation at an estimated 97 genetic loci. In addition to previously documented markers, variation was observed for the enzymes aconitase, aldehyde dehydrogenase, and alkaline phosphatase. The genetic basis of these markers (Acon-1, Ahd-2, and Akp-1) was confirmed. Linkage analysis between 35 pairwise comparisons revealed that the markers Fh-1 and Pep-3 are linked. The strain profiles of the 25 inbred strains at 11 electrophoretic markers are given.

Effects of gauge volume on pseudo-strain induced in strain measurement using time-of-flight neutron diffraction

International Nuclear Information System (INIS)

Suzuki, Hiroshi; Harjo, Stefanus; Abe, Jun; Xu, Pingguang; Aizawa, Kazuya; Akita, Koichi

2013-01-01

Spurious or pseudo-strains observed in time-of-flight (TOF) neutron diffraction due to neutron attenuation, surface-effects and a strain distribution within the gauge volume were investigated. Experiments were carried out on annealed and bent ferritic steel bars to test these effects. The most representative position in the gauge volume corresponds to the neutron-weighted center of gravity (ncog), which takes into account variations in intensity within the gauge volume due to neutron attenuation and/or absence of material in the gauge volume. The average strain in the gauge volume was observed to be weighted towards the ncog position but following an increase in the size of the gauge volume the weighted average strain was changed because of the change in the ncog position when a strain gradient appeared within the gauge volume. On the other hand, typical pseudo-strains, which are well known, did appear in through-surface strain measurements when the gauge volume was incompletely filled by the sample. Tensile pseudo-strains due to the surface-effect increased near the sample surface and exhibited a similar trend regardless of the size of the gauge volume, while the pseudo-strains increased faster for the smaller gauge volume. Furthermore, a pseudo-strain due to a change in the ncog position was observed even when the gauge volume was perfectly filled in the sample, and it increased with an increase in the size of the gauge volume. These pseudo-strains measured were much larger than those simulated by the conventional modeling, whereas they were simulated by taking into account an incident neutron beam divergence additionally in the model. Therefore, the incident divergence of the incident neutron beam must be carefully designed to avoid pseudo-strains in time-of-flight neutron diffractometry

A wide extent of inter-strain diversity in virulent and vaccine strains of alphaherpesviruses.

Science.gov (United States)

Szpara, Moriah L; Tafuri, Yolanda R; Parsons, Lance; Shamim, S Rafi; Verstrepen, Kevin J; Legendre, Matthieu; Enquist, L W

2011-10-01

Alphaherpesviruses are widespread in the human population, and include herpes simplex virus 1 (HSV-1) and 2, and varicella zoster virus (VZV). These viral pathogens cause epithelial lesions, and then infect the nervous system to cause lifelong latency, reactivation, and spread. A related veterinary herpesvirus, pseudorabies (PRV), causes similar disease in livestock that result in significant economic losses. Vaccines developed for VZV and PRV serve as useful models for the development of an HSV-1 vaccine. We present full genome sequence comparisons of the PRV vaccine strain Bartha, and two virulent PRV isolates, Kaplan and Becker. These genome sequences were determined by high-throughput sequencing and assembly, and present new insights into the attenuation of a mammalian alphaherpesvirus vaccine strain. We find many previously unknown coding differences between PRV Bartha and the virulent strains, including changes to the fusion proteins gH and gB, and over forty other viral proteins. Inter-strain variation in PRV protein sequences is much closer to levels previously observed for HSV-1 than for the highly stable VZV proteome. Almost 20% of the PRV genome contains tandem short sequence repeats (SSRs), a class of nucleic acids motifs whose length-variation has been associated with changes in DNA binding site efficiency, transcriptional regulation, and protein interactions. We find SSRs throughout the herpesvirus family, and provide the first global characterization of SSRs in viruses, both within and between strains. We find SSR length variation between different isolates of PRV and HSV-1, which may provide a new mechanism for phenotypic variation between strains. Finally, we detected a small number of polymorphic bases within each plaque-purified PRV strain, and we characterize the effect of passage and plaque-purification on these polymorphisms. These data add to growing evidence that even plaque-purified stocks of stable DNA viruses exhibit limited sequence

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

Directory of Open Access Journals (Sweden)

Hiroki Dan

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

Hydrogen production from microbial strains

Science.gov (United States)

Harwood, Caroline S; Rey, Federico E

2012-09-18

The present invention is directed to a method of screening microbe strains capable of generating hydrogen. This method involves inoculating one or more microbes in a sample containing cell culture medium to form an inoculated culture medium. The inoculated culture medium is then incubated under hydrogen producing conditions. Once incubating causes the inoculated culture medium to produce hydrogen, microbes in the culture medium are identified as candidate microbe strains capable of generating hydrogen. Methods of producing hydrogen using one or more of the microbial strains identified as well as the hydrogen producing strains themselves are also disclosed.

Bacteriocin-like inhibitory activities of seven Lactobacillus delbrueckii subsp. bulgaricus strains against antibiotic susceptible and resistant Helicobacter pylori strains.

Science.gov (United States)

Boyanova, L; Gergova, G; Markovska, R; Yordanov, D; Mitov, I

2017-12-01

The aim of the study was to detect anti-Helicobacter pylori activity of seven Lactobacillus delbrueckii subsp. bulgaricus (GLB) strains by four cell-free supernatant (CFS) types. Activity of non-neutralized and non-heat-treated (CFSs1), non-neutralized and heat-treated (CFSs2), pH neutralized, catalase-treated and non-heat-treated (CFSs3), or neutralized, catalase- and heat-treated (CFSs4) CFSs against 18 H. pylori strains (11 of which with antibiotic resistance) was evaluated. All GLB strains produced bacteriocin-like inhibitory substances (BLISs), the neutralized CFSs of two GLB strains inhibited >81% of test strains and those of four GLB strains were active against >71% of antibiotic resistant strains. Two H. pylori strains were BLIS resistant. The heating did not reduce the CFS activity. Briefly, all GLB strains evaluated produced heat-stable BLISs, although GLB and H. pylori strain susceptibility patterns exhibited differences. Bacteriocin-like inhibitory substance activity can be an advantage for the probiotic choice for H. pylori infection control. In this study, anti-Helicobacter pylori activity of seven Lactobacillus delbrueckii subsp. bulgaricus (GLB) strains was evaluated by four cell-free supernatant (CFS) types. The GLB strains produced heat-stable bacteriocin-like inhibitory substances (BLISs) with a strong anti-H. pylori activity and some neutralized, catalase- and heat-treated CFSs inhibited >83% of the test strains. Bacteriocin-like inhibitory substance production of GLB strains can render them valuable probiotics in the control of H. pylori infection. © 2017 The Society for Applied Microbiology.

Pin clad strains in Phenix

International Nuclear Information System (INIS)

Languille, A.

1979-07-01

The Phenix reactor has operated for 4 years in a satisfactory manner. The first 2 sub-assembly loadings contained pins clad in solution treated 316. The principal pin strains are: diametral strain (swelling and irradiation creep), ovality and spiral bending of the pin (interaction of wire and pin cluster and wrapper). A pin cluster irradiated to a dose of 80 dpa F reached a pin diameter strain of 5%. This strain is principally due to swelling (low fission gas pressure). The principal parameters governing the swelling are instantaneous dose, time and temperature for a given type of pin cladding. Other types of steel are or will be irradiated in Phenix. In particular, cold-worked titanium stabilised 316 steel should contribute towards a reduction in the pin clad strains and increase the target burn-up in this reactor. (author)

Strain screen and haplotype association mapping of wheel running in inbred mouse strains.

Science.gov (United States)

Lightfoot, J Timothy; Leamy, Larry; Pomp, Daniel; Turner, Michael J; Fodor, Anthony A; Knab, Amy; Bowen, Robert S; Ferguson, David; Moore-Harrison, Trudy; Hamilton, Alicia

2010-09-01

Previous genetic association studies of physical activity, in both animal and human models, have been limited in number of subjects and genetically homozygous strains used as well as number of genomic markers available for analysis. Expansion of the available mouse physical activity strain screens and the recently published dense single-nucleotide polymorphism (SNP) map of the mouse genome (approximately 8.3 million SNPs) and associated statistical methods allowed us to construct a more generalizable map of the quantitative trait loci (QTL) associated with physical activity. Specifically, we measured wheel running activity in male and female mice (average age 9 wk) in 41 inbred strains and used activity data from 38 of these strains in a haplotype association mapping analysis to determine QTL associated with activity. As seen previously, there was a large range of activity patterns among the strains, with the highest and lowest strains differing significantly in daily distance run (27.4-fold), duration of activity (23.6-fold), and speed (2.9-fold). On a daily basis, female mice ran further (24%), longer (13%), and faster (11%). Twelve QTL were identified, with three (on Chr. 12, 18, and 19) in both male and female mice, five specific to males, and four specific to females. Eight of the 12 QTL, including the 3 general QTL found for both sexes, fell into intergenic areas. The results of this study further support the findings of a moderate to high heritability of physical activity and add general genomic areas applicable to a large number of mouse strains that can be further mined for candidate genes associated with regulation of physical activity. Additionally, results suggest that potential genetic mechanisms arising from traditional noncoding regions of the genome may be involved in regulation of physical activity.

Temperature and strain registration by fibre-optic strain sensor in the polymer composite materials manufacturing

Science.gov (United States)

Matveenko; Kosheleva; Shardakov; Voronkov

2018-04-01

The presence of process-induced strains induced by various manufacturing and operational factors is one of the characteristics of polymer composite materials (PCM). Conventional methods of registration and evaluation of process-induced strains can be laborious, time-consuming and demanding in terms of technical applications. The employment of embedded fibre-optic strain sensors (FOSS) offers a real prospect of measuring residual strains. This paper demonstrates the potential for using embedded FOSS for recording technological strains in a PCM plate. The PCM plate is manufactured from prepreg, using the direct compression-moulding method. In this method, the prepared reinforcing package is placed inside a mould, heated, and then exposed to compaction pressure. The examined technology can be used for positioning FOSS between the layers of the composite material. Fibre-optic sensors, interacting with the material of the examined object, make it possible to register the evolution of the strain process during all stages of polymer-composite formation. FOSS data were recorded with interrogator ASTRO X 327. The obtained data were processed using specially developed algorithms.

Sensitivity Enhancement of FBG-Based Strain Sensor.

Science.gov (United States)

Li, Ruiya; Chen, Yiyang; Tan, Yuegang; Zhou, Zude; Li, Tianliang; Mao, Jian

2018-05-17

A novel fiber Bragg grating (FBG)-based strain sensor with a high-sensitivity is presented in this paper. The proposed FBG-based strain sensor enhances sensitivity by pasting the FBG on a substrate with a lever structure. This typical mechanical configuration mechanically amplifies the strain of the FBG to enhance overall sensitivity. As this mechanical configuration has a high stiffness, the proposed sensor can achieve a high resonant frequency and a wide dynamic working range. The sensing principle is presented, and the corresponding theoretical model is derived and validated. Experimental results demonstrate that the developed FBG-based strain sensor achieves an enhanced strain sensitivity of 6.2 pm/με, which is consistent with the theoretical analysis result. The strain sensitivity of the developed sensor is 5.2 times of the strain sensitivity of a bare fiber Bragg grating strain sensor. The dynamic characteristics of this sensor are investigated through the finite element method (FEM) and experimental tests. The developed sensor exhibits an excellent strain-sensitivity-enhancing property in a wide frequency range. The proposed high-sensitivity FBG-based strain sensor can be used for small-amplitude micro-strain measurement in harsh industrial environments.

Haemophilus ducreyi Cutaneous Ulcer Strains Diverged from Both Class I and Class II Genital Ulcer Strains: Implications for Epidemiological Studies.

Directory of Open Access Journals (Sweden)

Dharanesh Gangaiah

2016-12-01

Full Text Available Haemophilus ducreyi has emerged as a major cause of cutaneous ulcers (CU in yaws-endemic regions of the tropics in the South Pacific, South East Asia and Africa. H. ducreyi was once thought only to cause the genital ulcer (GU disease chancroid; GU strains belong to 2 distinct classes, class I and class II. Using whole-genome sequencing of 4 CU strains from Samoa, 1 from Vanuatu and 1 from Papua New Guinea, we showed that CU strains diverged from the class I strain 35000HP and that one CU strain expressed β-lactamase. Recently, the Center for Disease Control and Prevention released the genomes of 11 additional CU strains from Vanuatu and Ghana; however, the evolutionary relationship of these CU strains to previously-characterized CU and GU strains is unknown.We performed phylogenetic analysis of 17 CU and 10 GU strains. Class I and class II GU strains formed two distinct clades. The class I strains formed two subclades, one containing 35000HP and HD183 and the other containing the remainder of the class I strains. Twelve of the CU strains formed a subclone under the class I 35000HP subclade, while 2 CU strains formed a subclone under the other class I subclade. Unexpectedly, 3 of the CU strains formed a subclone under the class II clade. Phylogenetic analysis of dsrA-hgbA-ncaA sequences yielded a tree similar to that of whole-genome phylogenetic tree.CU strains diverged from multiple lineages within both class I and class II GU strains. Multilocus sequence typing of dsrA-hgbA-ncaA could be reliably used for epidemiological investigation of CU and GU strains. As class II strains grow relatively poorly and are relatively more susceptible to vancomycin than class I strains, these findings have implications for methods to recover CU strains. Comparison of contemporary CU and GU isolates would help clarify the relationship between these entities.

Haemophilus ducreyi Cutaneous Ulcer Strains Diverged from Both Class I and Class II Genital Ulcer Strains: Implications for Epidemiological Studies.

Science.gov (United States)

Gangaiah, Dharanesh; Spinola, Stanley M

2016-12-01

Haemophilus ducreyi has emerged as a major cause of cutaneous ulcers (CU) in yaws-endemic regions of the tropics in the South Pacific, South East Asia and Africa. H. ducreyi was once thought only to cause the genital ulcer (GU) disease chancroid; GU strains belong to 2 distinct classes, class I and class II. Using whole-genome sequencing of 4 CU strains from Samoa, 1 from Vanuatu and 1 from Papua New Guinea, we showed that CU strains diverged from the class I strain 35000HP and that one CU strain expressed β-lactamase. Recently, the Center for Disease Control and Prevention released the genomes of 11 additional CU strains from Vanuatu and Ghana; however, the evolutionary relationship of these CU strains to previously-characterized CU and GU strains is unknown. We performed phylogenetic analysis of 17 CU and 10 GU strains. Class I and class II GU strains formed two distinct clades. The class I strains formed two subclades, one containing 35000HP and HD183 and the other containing the remainder of the class I strains. Twelve of the CU strains formed a subclone under the class I 35000HP subclade, while 2 CU strains formed a subclone under the other class I subclade. Unexpectedly, 3 of the CU strains formed a subclone under the class II clade. Phylogenetic analysis of dsrA-hgbA-ncaA sequences yielded a tree similar to that of whole-genome phylogenetic tree. CU strains diverged from multiple lineages within both class I and class II GU strains. Multilocus sequence typing of dsrA-hgbA-ncaA could be reliably used for epidemiological investigation of CU and GU strains. As class II strains grow relatively poorly and are relatively more susceptible to vancomycin than class I strains, these findings have implications for methods to recover CU strains. Comparison of contemporary CU and GU isolates would help clarify the relationship between these entities.

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

Science.gov (United States)

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

2013-01-01

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

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

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.

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.

Strain quantification in epitaxial thin films

International Nuclear Information System (INIS)

Cushley, M

2008-01-01

Strain arising in epitaxial thin films can be beneficial in some cases but devastating in others. By altering the lattice parameters, strain may give a thin film properties hitherto unseen in the bulk material. On the other hand, heavily strained systems are prone to develop lattice defects in order to relieve the strain, which can cause device failure or, at least, a decrease in functionality. Using convergent beam electron diffraction (CBED) and high-resolution transmission electron microscopy (HRTEM), it is possible to determine local strains within a material. By comparing the results from CBED and HRTEM experiments, it is possible to gain a complete view of a material, including the strain and any lattice defects present. As well as looking at how the two experimental techniques differ from each other, I will also look at how results from different image analysis algorithms compare. Strain in Si/SiGe samples and BST/SRO/MgO capacitor structures will be discussed.

Brucella abortus strain 2308 Wisconsin genome: importance of the definition of reference strains

Directory of Open Access Journals (Sweden)

Marcela Suárez-Esquivel

2016-09-01

Full Text Available Brucellosis is a bacterial infectious disease affecting a wide range of mammals and a neglected zoonosis caused by species of the genetically homogenous genus Brucella. As in most studies on bacterial diseases, research in brucellosis is carried out by using reference strains as canonical models to understand the mechanisms underlying host pathogen interactions. We performed whole genome sequencing (WGS analysis of the reference strain Brucella abortus 2308 routinely used in our laboratory, including manual curated annotation accessible as an editable version at www.wikipedia.Comparison of this genome with two publically available 2308 genomes showed significant differences, particularly indels related to insertional elements, suggesting variability related to the transposition of these elements within the same strain. Considering the outcome of high resolution genomic techniques in the bacteriology field, the conventional concept of strain definition needs to be revised.

Molecular characterization of the probiotic strain Bacillus cereus var. toyoi NCIMB 40112 and differentiation from food poisoning strains.

Science.gov (United States)

Klein, Günter

2011-07-01

Bacillus cereus var. toyoi strain NCIMB 40112 (Toyocerin), a probiotic authorized in the European Union as feed additive for swine, bovines, poultry, and rabbits, was characterized by DNA fingerprinting applying pulsed-field gel electrophoresis and multilocus sequence typing and was compared with reference strains (of clinical and environmental origins). The probiotic strain was clearly characterized by pulsed-field gel electrophoresis using the restriction enzymes Apa I and Sma I resulting in unique DNA patterns. The comparison to the clinical reference strain B. cereus DSM 4312 was done with the same restriction enzymes, and again a clear differentiation of the two strains was possible by the resulting DNA patterns. The use of the restriction enzymes Apa I and Sma I is recommended for further studies. Furthermore, multilocus sequence typing analysis revealed a sequence type (ST 111) that was different from all known STs of B. cereus strains from food poisoning incidents. Thus, a strain characterization and differentiation from food poisoning strains for the probiotic strain was possible. Copyright ©, International Association for Food Protection

Microstructure and strain distribution in freestanding Si membrane strained by Si{sub x}N{sub y} deposition

Energy Technology Data Exchange (ETDEWEB)

Gao Hongye, E-mail: qgaohongye@msn.com [Faculty of Engineering Sciences, Kyushu University, 6-1 Kasuga-koen, Kasuga, Fukuoka 816-8580 (Japan); Ikeda, Ken-ichi; Hata, Satoshi; Nakashima, Hideharu [Faculty of Engineering Sciences, Kyushu University, 6-1 Kasuga-koen, Kasuga, Fukuoka 816-8580 (Japan); Wang Dong; Nakashima, Hiroshi [Art, Science and Technology Center for Cooperative Research, Kyushu University, 6-1 Kasuga-koen, Kasuga, Fukuoka 816-8580 (Japan)

2010-09-25

Research highlights: {yields} Strain is introduced by deposition of amorphous Si{sub x}N{sub y} to improve the carrier mobility for a relatively large-size freestanding semiconductor film, which can be used for the fabrication of relatively large devices such like a bipolar junction transistor. However, standard Raman spectroscopy and X-ray diffraction cannot provide sufficient lateral resolution to the strain in a relatively long (x {mu}m in length) and thin (x nm in thickness) freestanding semiconductor film. {yields} In present research, strain in a bridge-shaped freestanding Si membrane (FSSM) was measured by convergent-beam electron diffraction (CBED) and finite element method (FEM). Compressive strain distribution was shown in three dimensions (3D) in FSSM, where no threading dislocation or stacking fault was found. Relaxation of the strain in FSSM in 3D was discussed based on a comparison of the strain magnitudes in FSSM as measured by CBED and FEM. - Abstract: Strain in a bridge-shaped freestanding Si membrane (FSSM) induced by depositing an amorphous Si{sub x}N{sub y} layer was measured by convergent-beam electron diffraction (CBED). CBED results show that the strain magnitude depends negatively on the FSSM thickness. FEM is a supplement of the result of CBED due to the relaxation of TEM samples during fabricating. The FEM analysis results ascertain the strain property in three dimensions, and show that the strain magnitude depends negatively on the length of FSSM, and the magnitude of the compressive strain in FSSM increases as the position is closer to the upper Si/Si{sub x}N{sub y} interface.

Genomics-enabled analysis of the emergent disease cotton bacterial blight.

Directory of Open Access Journals (Sweden)

Anne Z Phillips

2017-09-01

Full Text Available Cotton bacterial blight (CBB, an important disease of (Gossypium hirsutum in the early 20th century, had been controlled by resistant germplasm for over half a century. Recently, CBB re-emerged as an agronomic problem in the United States. Here, we report analysis of cotton variety planting statistics that indicate a steady increase in the percentage of susceptible cotton varieties grown each year since 2009. Phylogenetic analysis revealed that strains from the current outbreak cluster with race 18 Xanthomonas citri pv. malvacearum (Xcm strains. Illumina based draft genomes were generated for thirteen Xcm isolates and analyzed along with 4 previously published Xcm genomes. These genomes encode 24 conserved and nine variable type three effectors. Strains in the race 18 clade contain 3 to 5 more effectors than other Xcm strains. SMRT sequencing of two geographically and temporally diverse strains of Xcm yielded circular chromosomes and accompanying plasmids. These genomes encode eight and thirteen distinct transcription activator-like effector genes. RNA-sequencing revealed 52 genes induced within two cotton cultivars by both tested Xcm strains. This gene list includes a homeologous pair of genes, with homology to the known susceptibility gene, MLO. In contrast, the two strains of Xcm induce different clade III SWEET sugar transporters. Subsequent genome wide analysis revealed patterns in the overall expression of homeologous gene pairs in cotton after inoculation by Xcm. These data reveal important insights into the Xcm-G. hirsutum disease complex and strategies for future development of resistant cultivars.

The asymptomatic bacteriuria Escherichia coli strain 83972 outcompetes uropathogenic E. coli strains in human urine

DEFF Research Database (Denmark)

Hancock, Viktoria; Ulett, G.C.; Schembri, M.A.

2006-01-01

Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU). In contrast to uropathogenic E. coli (UPEC), which causes symptomatic urinary tract infections (UTI), very little is known about the mechanisms by which these strains colonize the human urinary tract....... The prototype ABU E. coli strain 83972 was originally isolated from a girl who had carried it asymptomatically for 3 years. Deliberate colonization of UTI-susceptible individuals with E. coli 83972 has been used successfully as an alternative approach for the treatment of patients who are refractory...... to conventional therapy. Colonization with strain 83972 appears to prevent infection with UPEC strains in such patients despite the fact that this strain is unable to express the primary adhesins involved in UTI, viz. P and type 1 fimbriae. Here we investigated the growth characteristics of E. coli 83972 in human...

Three Phoma spp. synthesised novel silver nanoparticles that possess excellent antimicrobial efficacy.

Science.gov (United States)

Rai, Mahendra; Ingle, Avinash P; Gade, Aniket K; Duarte, Marta Cristina Teixeira; Duran, Nelson

2015-10-01

The authors report extracellular mycosynthesis of silver nanoparticles (AgNPs) by Phoma capsulatum, Phoma putaminum and Phoma citri. The AgNPs thus synthesised were characterised by UV-visible spectrophotometer, Fourier transform infrared spectroscopy, Nanosight LM20 and transmission electron microscopy, which confirmed the synthesis of mostly spherical and polydisperse nanoparticles capped with proteins. The size of AgNPs was found in the range of 10-80 , 5-80 and 5-90 nm with an average size of 31.85, 25.43 and 23.29 nm by P. capsulatum, P. putaminum and P. citri, respectively. Further, potential antimicrobial activity was reported against Aspergillus niger, Candida albicans, Salmonella choleraesuis, Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli. The lowest minimal inhibitory concentration (MIC) (0.85 µg/ml) was reported for AgNPs synthesised from P. citri against S. choleraesuis. However, AgNPs synthesised from P. capsulatum showed the highest MIC (10.62 µg/ml) against S. choleraesuis, P. aeruginosa and E. coli (clinical isolate). The same MIC values (10.62 µg/ml) were also reported against P. aeruginosa and both clinical and standard isolates of E. coli for AgNPs synthesised from P. citri. It was also observed that all the silver nanoparticles showed remarkable antifungal and antibacterial activity against these tested pathogens as compared with the commercially available antifungal and antibacterial agents.

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.

Acquirement of true stress-strain curve using true fracture strain obtained by tensile test and FE analysis

International Nuclear Information System (INIS)

Lee, Kyoung Yoon; Kim, Tae Hyung; Lee, Hyung Yil

2009-01-01

In this work, we predict a true fracture strain using load-displacement curves from tensile test and Finite Element Analysis (FEA), and suggest a method for acquiring true Stress-Strain (SS) curves by predicted fracture strain. We first derived the true SS curve up to necking point from load-displacement curve. As the beginning, the posterior necking part of true SS curve is linearly extrapolated with the slope at necking point. The whole SS curve is then adopted for FE simulation of tensile test. The Bridgman factor or suitable plate correction factors are applied to pre and post FEA. In the load-true strain curve from FEA, the true fracture strain is determined as the matching point to test fracture load. The determined true strain is validated by comparing with test fracture strain. Finally, we complete the true SS curve by combining the prior necking part and linear part, the latter of which connects necking and predicted fracture points.

Acquirement of true stress-strain curve using true fracture strain obtained by tensile test and FE analysis

Energy Technology Data Exchange (ETDEWEB)

Lee, Kyoung Yoon; Kim, Tae Hyung; Lee, Hyung Yil [Sogang University, Seoul (Korea, Republic of)

2009-07-01

In this work, we predict a true fracture strain using load-displacement curves from tensile test and Finite Element Analysis (FEA), and suggest a method for acquiring true Stress-Strain (SS) curves by predicted fracture strain. We first derived the true SS curve up to necking point from load-displacement curve. As the beginning, the posterior necking part of true SS curve is linearly extrapolated with the slope at necking point. The whole SS curve is then adopted for FE simulation of tensile test. The Bridgman factor or suitable plate correction factors are applied to pre and post FEA. In the load-true strain curve from FEA, the true fracture strain is determined as the matching point to test fracture load. The determined true strain is validated by comparing with test fracture strain. Finally, we complete the true SS curve by combining the prior necking part and linear part, the latter of which connects necking and predicted fracture points.

Acquirement of True Stress-strain Curve Using True Fracture Strain Obtained by Tensile Test and FE Analysis

Energy Technology Data Exchange (ETDEWEB)

Lee, Kyoung Yoon; Lee, Hyung Yil [Sogang University, Seoul (Korea, Republic of); Kim, Tae Hyung [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

2009-10-15

In this work, we predict a true fracture strain using load-displacement curves from tensile test and finite element analysis (FEA), and suggest a method for acquiring true stress-strain (SS) curves by predicted fracture strain. We first derived the true SS curve up to necking point from load-displacement curve. As the beginning, the posterior necking part of true SS curve is linearly extrapolated with the slope at necking point. The whole SS curve is then adopted for FE simulation of tensile test. The Bridgman factor or suitable plate correction factors are applied to pre and post FEA. In the load-true strain curve from FEA, the true fracture strain is determined as the matching point to test fracture load. The determined true strain is validated by comparing with test fracture strain. Finally, we complete the true SS curve by combining the prior necking part and linear part, the latter of which connects necking and predicted fracture points.

Atlas of stress-strain curves

CERN Document Server

2002-01-01

The Atlas of Stress-Strain Curves, Second Edition is substantially bigger in page dimensions, number of pages, and total number of curves than the previous edition. It contains over 1,400 curves, almost three times as many as in the 1987 edition. The curves are normalized in appearance to aid making comparisons among materials. All diagrams include metric (SI) units, and many also include U.S. customary units. All curves are captioned in a consistent format with valuable information including (as available) standard designation, the primary source of the curve, mechanical properties (including hardening exponent and strength coefficient), condition of sample, strain rate, test temperature, and alloy composition. Curve types include monotonic and cyclic stress-strain, isochronous stress-strain, and tangent modulus. Curves are logically arranged and indexed for fast retrieval of information. The book also includes an introduction that provides background information on methods of stress-strain determination, on...

Graphene spin diode: Strain-modulated spin rectification

Energy Technology Data Exchange (ETDEWEB)

Wang, Yunhua; Wang, B., E-mail: stslyl@mail.sysu.edu.cn, E-mail: wangbiao@mail.sysu.edu.cn [Sino-French Institute of Nuclear Engineering and Technology, School of Physics and Engineering, State Key Laboratory of Optoelectronic Materials and Technologies, Sun Yat-sen University, Guangzhou 510275 (China); Liu, Yulan, E-mail: stslyl@mail.sysu.edu.cn, E-mail: wangbiao@mail.sysu.edu.cn [School of Engineering, Sun Yat-sen University, Guangzhou 510275 (China)

2014-08-04

Strain effects on spin transport in a ferromagnetic/strained/normal graphene junction are explored theoretically. It is shown that the spin-resolved Fermi energy range can be controlled by the armchair direction strain because the strain-induced pseudomagnetic field suppresses the current. The spin rectification effect for the bias reversal occurs because of a combination of ferromagnetic exchange splitting and the broken spatial symmetry of the junction. In addition, the spin rectification performance can be tuned remarkably by manipulation of the strains. In view of this strain-modulated spin rectification effect, we propose that the graphene-based ferromagnetic/strained/normal junction can be used as a tunable spin diode.

Development of Industrial Yeast Platform Strains

DEFF Research Database (Denmark)

Bergdahl, Basti; Dato, Laura; Förster, Jochen

2014-01-01

Most of the current metabolic engineering projects are carried out using laboratory strains as the starting host. Although such strains are easily manipulated genetically, their robustness does not always meet the requirements set by industrial fermentation conditions. In such conditions, the cells...... screening of the 36 industrial and laboratory yeast strains. In addition, progress in the development of molecular biology methods for generating the new strains will be presented....

Taxonomic and Strain-Specific Identification of the Probiotic Strain Lactobacillus rhamnosus 35 within the Lactobacillus casei Group▿

Science.gov (United States)

Coudeyras, Sophie; Marchandin, Hélène; Fajon, Céline; Forestier, Christiane

2008-01-01

Lactobacilli are lactic acid bacteria that are widespread in the environment, including the human diet and gastrointestinal tract. Some Lactobacillus strains are regarded as probiotics because they exhibit beneficial health effects on their host. In this study, the long-used probiotic strain Lactobacillus rhamnosus 35 was characterized at a molecular level and compared with seven reference strains from the Lactobacillus casei group. Analysis of rrn operon sequences confirmed that L. rhamnosus 35 indeed belongs to the L. rhamnosus species, and both temporal temperature gradient gel electrophoresis and ribotyping showed that it is closer to the probiotic strain L. rhamnosus ATCC 53103 (also known as L. rhamnosus GG) than to the species type strain. In addition, L. casei ATCC 334 gathered in a coherent cluster with L. paracasei type strains, unlike L. casei ATCC 393, which was closer to L. zeae; this is evidence of the lack of relatedness between the two L. casei strains. Further characterization of the eight strains by pulsed-field gel electrophoresis repetitive DNA element-based PCR identified distinct patterns for each strain, whereas two isolates of L. rhamnosus 35 sampled 40 years apart could not be distinguished. By subtractive hybridization using the L. rhamnosus GG genome as a driver, we were able to isolate five L. rhamnosus 35-specific sequences, including two phage-related ones. The primer pairs designed to amplify these five regions allowed us to develop rapid and highly specific PCR-based identification methods for the probiotic strain L. rhamnosus 35. PMID:18326671

Brucella abortus Strain 2308 Wisconsin Genome: Importance of the Definition of Reference Strains

Science.gov (United States)

Suárez-Esquivel, Marcela; Ruiz-Villalobos, Nazareth; Castillo-Zeledón, Amanda; Jiménez-Rojas, César; Roop II, R. Martin; Comerci, Diego J.; Barquero-Calvo, Elías; Chacón-Díaz, Carlos; Caswell, Clayton C.; Baker, Kate S.; Chaves-Olarte, Esteban; Thomson, Nicholas R.; Moreno, Edgardo; Letesson, Jean J.; De Bolle, Xavier; Guzmán-Verri, Caterina

2016-01-01

Brucellosis is a bacterial infectious disease affecting a wide range of mammals and a neglected zoonosis caused by species of the genetically homogenous genus Brucella. As in most studies on bacterial diseases, research in brucellosis is carried out by using reference strains as canonical models to understand the mechanisms underlying host pathogen interactions. We performed whole genome sequencing analysis of the reference strain B. abortus 2308 routinely used in our laboratory, including manual curated annotation accessible as an editable version through a link at https://en.wikipedia.org/wiki/Brucella#Genomics. Comparison of this genome with two publically available 2308 genomes showed significant differences, particularly indels related to insertional elements, suggesting variability related to the transposition of these elements within the same strain. Considering the outcome of high resolution genomic techniques in the bacteriology field, the conventional concept of strain definition needs to be revised. PMID:27746773

Different distribution patterns of ten virulence genes in Legionella reference strains and strains isolated from environmental water and patients.

Science.gov (United States)

Zhan, Xiao-Yong; Hu, Chao-Hui; Zhu, Qing-Yi

2016-04-01

Virulence genes are distinct regions of DNA which are present in the genome of pathogenic bacteria and absent in nonpathogenic strains of the same or related species. Virulence genes are frequently associated with bacterial pathogenicity in genus Legionella. In the present study, an assay was performed to detect ten virulence genes, including iraA, iraB, lvrA, lvrB, lvhD, cpxR, cpxA, dotA, icmC and icmD in different pathogenicity islands of 47 Legionella reference strains, 235 environmental strains isolated from water, and 4 clinical strains isolated from the lung tissue of pneumonia patients. The distribution frequencies of these genes in reference or/and environmental L. pneumophila strains were much higher than those in reference non-L. pneumophila or/and environmental non-L. pneumophila strains, respectively. L. pneumophila clinical strains also maintained higher frequencies of these genes compared to four other types of Legionella strains. Distribution frequencies of these genes in reference L. pneumophila strains were similar to those in environmental L. pneumophila strains. In contrast, environmental non-L. pneumophila maintained higher frequencies of these genes compared to those found in reference non-L. pneumophila strains. This study illustrates the association of virulence genes with Legionella pathogenicity and reveals the possible virulence evolution of non-L. pneumophia strains isolated from environmental water.

Development of high temperature strain gage, (5)

International Nuclear Information System (INIS)

Yuuki, Hiroshi; Kobayashi, Yukio; Kanai, Kenji; Yamaura, Yoshio

1976-01-01

Development and improvement of resistance wire type strain gages usable for experimental measurement of thermal strains generated at high temperature in various structures and equipments that consist of a Fast Breeder Reactor have been carried out, and various characteristics of the strain gages have been investigated. Based on the results obtained up to now, development and research of this time mainly aim to improve strain and fatigue characteristics. As the results, characteristics of strain gages with sensing elements of nichrome V are improved, specifically mechanical hysteresis is decreased, strain limit is increased, etc. Also, improvement is recognized in thermal output, and it becomes clear that dummy gages work effectively. However, a filling method of MgO and an inserting method of active-dummy elements are selected as primary objects to improve strain characteristics, and many hours are taken for these objects, so confirmations of characteristics of platinum-tungsten strain gages, strain sensing elements of which are troublesome to produce, have not been completely done, though the performance of the gages has been improved in several points. As to nichrome V strain gages, there is a fair prospect of obtaining ones, specifications of which are quite close to the goal, though problems in manufacturing technics remain for future. As to platinum-tungsten strain gages, it is expected that similar strain gages to nichrome V are obtainable by improvement in manufacturing of sensing elements. (auth.)

Differentiation of five strains of infectious bursal disease virus: Development of a strain-specific multiplex PCR

DEFF Research Database (Denmark)

Kusk, M.; Kabell, Susanne; Jørgensen, Poul Henrik

2005-01-01

and histopathology. Since these methods are laborious and have low specificity alternatives are needed. In the present study, we report the development of a strain-specific multiplex RT-PCR technique, which can detect and differentiate between field strains of IBDV and vaccine virus strains including a so-called hot...

The effect of nutritional spray programs applied to mitigate symptoms of Huanglongbing on fruit drop caused by HLB and citrus canker on ‘Hamlin’ orange trees

OpenAIRE

Roberts, P. D.; Rouse, R. E.; Teems, S. S.; Sytsma, R. E.; Shobert, Z.

2014-01-01

Huanglongbing (HLB) was detected in Florida in 2005 and has reached 100% incidence in certain citrus plantings in southwest Florida. The putative causal agent of HLB in Florida is the bacterium Candidatus Liberibacter asiaticus (CLa).  Citrus canker caused by the bacterium Xanthomonas citri subsp. citri is endemic in Florida.  In 2011 and 2012, fruit drop on young ‘Hamlin’ trees with symptoms of HLB and/or citrus canker was particularly severe, with more than 90% fruit drop recorded. Nutritio...

MM98.57 Quantification of Combined Strain Paths

DEFF Research Database (Denmark)

Nielsen, Morten Sturgård; Wanheim, Tarras

1998-01-01

this curve into useful scalar relations from experimental data.The strain history for plane strain when assuming volume constancy may be plotted in a shear strain, normal strain diagram, which has the property of showing both the rotation of principal deformation axes during the deformation and the amount...... is to describe the total strain history as a curve in the 6-dimensional shear strain, normal strain space. In order to be able to use these experimental data for calculation, the development of this strain curve must be transformed into a set of scalar relations that may be used for predicting the yield surface...... at a given point in a new strain history. A simple example of this concept is to take the length of the strain curve as describing scalar relation: E.g. to use the equivalent strain as parameter for describing the yield stress. This paper focuses on the strain curve concept and the possibilities to convert...

Job strain and male fertility.

Science.gov (United States)

Hjollund, Niels Henrik I; Bonde, Jens Peter E; Henriksen, Tine Brink; Giwercman, Aleksander; Olsen, Jørn

2004-01-01

Job strain, defined as high job demands and low job control, has not previously been explored as a possible determinant of male fertility. We collected prospective data on job strain among men, and describe the associations with semen quality and probability of conceiving a clinical pregnancy during a menstrual cycle. Danish couples (N = 399) who were trying to become pregnant for the first time were followed for up to 6 menstrual periods. All men collected semen samples, and a blood sample was drawn from both partners. Job demand and job control were measured by a self-administered questionnaire at entry, and in each cycle the participants recorded changes in job control or job demand during the previous 30 days. In adjusted analyses, no associations were found between any semen characteristic or sexual hormones and any job strain variable. The odds for pregnancy were not associated with job strain. Psychologic job strain encountered in normal jobs in Denmark does not seem to affect male reproductive function.

Improved thermal stability and hole mobilities in a strained-Si/strained-Si1-yGe y/strained-Si heterostructure grown on a relaxed Si1-xGe x buffer

International Nuclear Information System (INIS)

Gupta, Saurabh; Lee, Minjoo L.; Isaacson, David M.; Fitzgerald, Eugene A.

2005-01-01

A dual channel heterostructure consisting of strained-Si/strained-Si 1-y Ge y on relaxed Si 1-x Ge x (y > x), provides a platform for fabricating metal-oxide-semiconductor field-effect transistors (MOSFETs) with high hole mobilities (μ eff ) which depend directly on Ge concentration and strain in the strained-Si 1-y Ge y layer. Ge out-diffuses from the strained-Si 1-y Ge y layer into relaxed Si 1-x Ge x during high temperature processing, reducing peak Ge concentration and strain in the strained-Si 1-y Ge y layer and degrades hole μ eff in these dual channel heterostructures. A heterostructure consisting of strained-Si/strained-Si 1-y Ge y /strained-Si, referred to as a trilayer heterostructure, grown on relaxed Si 1-x Ge x has much reduced Ge out-flux from the strained-Si 1-y Ge y layer and retains higher μ eff after thermal processing. Improved hole μ eff over similar dual channel heterostructures is also observed in this heterostructure. This could be a result of preventing the hole wavefunction tunneling into the low μ eff relaxed Si 1-x Ge x layer due to the additional valence band offset provided by the underlying strained-Si layer. A diffusion coefficient has been formulated and implemented in a finite difference scheme for predicting the thermal budget of the strained SiGe heterostructures. It shows that the trilayer heterostructures have superior thermal budgets at higher Ge concentrations. Ring-shaped MOSFETs were fabricated on both platforms and subjected to various processing temperatures in order to compare the extent of μ eff reduction with thermal budget. Hole μ eff enhancements are retained to a much higher extent in a trilayer heterostructure after high temperature processing as compared to a dual channel heterostructure. The improved thermal stability and hole μ eff of a trilayer heterostructure makes it an ideal platform for fabricating high μ eff MOSFETs that can be processed over higher temperatures without significant losses in hole �

Predicting creep rupture from early strain data

International Nuclear Information System (INIS)

Holmstroem, Stefan; Auerkari, Pertti

2009-01-01

To extend creep life modelling from classical rupture modelling, a robust and effective parametric strain model has been developed. The model can reproduce with good accuracy all parts of the creep curve, economically utilising the available rupture models. The resulting combined model can also be used to predict rupture from the available strain data, and to further improve the rupture models. The methodology can utilise unfailed specimen data for life assessment at lower stress levels than what is possible from rupture data alone. Master curves for creep strain and rupture have been produced for oxygen-free phosphorus-doped (OFP) copper with a maximum testing time of 51,000 h. Values of time to specific strain at given stress (40-165 MPa) and temperature (125-350 deg. C) were fitted to the models in the strain range of 0.1-38%. With typical inhomogeneous multi-batch creep data, the combined strain and rupture modelling involves the steps of investigation of the data quality, extraction of elastic and creep strain response, rupture modelling, data set balancing and creep strain modelling. Finally, the master curves for strain and rupture are tested and validated for overall fitting efficiency. With the Wilshire equation as the basis for the rupture model, the strain model applies classical parametric principles with an Arrhenius type of thermal activation and a power law type of stress dependence for the strain rate. The strain model also assumes that the processes of primary and secondary creep can be reasonably correlated. The rupture model represents a clear improvement over previous models in the range of the test data. The creep strain information from interrupted and running tests were assessed together with the rupture data investigating the possibility of rupture model improvement towards lower stress levels by inverse utilisation of the combined rupture based strain model. The developed creep strain model together with the improved rupture model is

A wide extent of inter-strain diversity in virulent and vaccine strains of alphaherpesviruses.

Directory of Open Access Journals (Sweden)

Moriah L Szpara

2011-10-01

Full Text Available Alphaherpesviruses are widespread in the human population, and include herpes simplex virus 1 (HSV-1 and 2, and varicella zoster virus (VZV. These viral pathogens cause epithelial lesions, and then infect the nervous system to cause lifelong latency, reactivation, and spread. A related veterinary herpesvirus, pseudorabies (PRV, causes similar disease in livestock that result in significant economic losses. Vaccines developed for VZV and PRV serve as useful models for the development of an HSV-1 vaccine. We present full genome sequence comparisons of the PRV vaccine strain Bartha, and two virulent PRV isolates, Kaplan and Becker. These genome sequences were determined by high-throughput sequencing and assembly, and present new insights into the attenuation of a mammalian alphaherpesvirus vaccine strain. We find many previously unknown coding differences between PRV Bartha and the virulent strains, including changes to the fusion proteins gH and gB, and over forty other viral proteins. Inter-strain variation in PRV protein sequences is much closer to levels previously observed for HSV-1 than for the highly stable VZV proteome. Almost 20% of the PRV genome contains tandem short sequence repeats (SSRs, a class of nucleic acids motifs whose length-variation has been associated with changes in DNA binding site efficiency, transcriptional regulation, and protein interactions. We find SSRs throughout the herpesvirus family, and provide the first global characterization of SSRs in viruses, both within and between strains. We find SSR length variation between different isolates of PRV and HSV-1, which may provide a new mechanism for phenotypic variation between strains. Finally, we detected a small number of polymorphic bases within each plaque-purified PRV strain, and we characterize the effect of passage and plaque-purification on these polymorphisms. These data add to growing evidence that even plaque-purified stocks of stable DNA viruses exhibit

[Characteristics of Lactobacillus strains contained in pharmaceuticals].

Science.gov (United States)

Banach, W; Bucholc, B; Wójcik, B

2001-01-01

The aim of the study was to characterize lactic acid bacteria (LAB) which are components of drugs administered orally in cases of intestinal disturbances, or antibiotic--related diarrhea. Biochemical properties, growth behavior, bile tolerance, and survival at low pH of six LAB strains (four strains L. rhamnosus and two L. acidophilus) were studied. The survival at low pH was determined in MRS broth (Difco) acidified to pH 1; 2; 3; and 4. Bile tolerance was tested on MRS broth with 0.3% oxgall (Difco). Between tested strains differences in ability to grow at low pH and survival in bile were observed. Only 0.01% inoculum of all examined strains survived at pH 1. Differences between strains in survival at low pH (pH 2 and pH 3) and tolerance of bile were observed. However, after 2 h incubation at pH 4, 100% of strains stayed alive. Examined strains demonstrated good 3% bile tolerance. All strains met the criteria for probiotic strains: ability to survive at pH 3 and in the presence of bile.

Strain histograms are equal to strain ratios in predicting malignancy in breast tumours

DEFF Research Database (Denmark)

Carlsen, Jonathan Frederik; Ewertsen, Caroline; Sletting, Susanne

2017-01-01

Objectives: To assess whether strain histograms are equal to strain ratios in predicting breast tumour malignancy and to see if either could be used to upgrade Breast Imaging Reporting and Data System (BI-RADS) 3 tumours for immediate biopsy. Methods: Ninety-nine breast tumours were examined using...

Strains and Stressors: An Analysis of Touchscreen Learning in Genetically Diverse Mouse Strains

Science.gov (United States)

Graybeal, Carolyn; Bachu, Munisa; Mozhui, Khyobeni; Saksida, Lisa M.; Bussey, Timothy J.; Sagalyn, Erica; Williams, Robert W.; Holmes, Andrew

2014-01-01

Touchscreen-based systems are growing in popularity as a tractable, translational approach for studying learning and cognition in rodents. However, while mouse strains are well known to differ in learning across various settings, performance variation between strains in touchscreen learning has not been well described. The selection of appropriate genetic strains and backgrounds is critical to the design of touchscreen-based studies and provides a basis for elucidating genetic factors moderating behavior. Here we provide a quantitative foundation for visual discrimination and reversal learning using touchscreen assays across a total of 35 genotypes. We found significant differences in operant performance and learning, including faster reversal learning in DBA/2J compared to C57BL/6J mice. We then assessed DBA/2J and C57BL/6J for differential sensitivity to an environmental insult by testing for alterations in reversal learning following exposure to repeated swim stress. Stress facilitated reversal learning (selectively during the late stage of reversal) in C57BL/6J, but did not affect learning in DBA/2J. To dissect genetic factors underlying these differences, we phenotyped a family of 27 BXD strains generated by crossing C57BL/6J and DBA/2J. There was marked variation in discrimination, reversal and extinction learning across the BXD strains, suggesting this task may be useful for identifying underlying genetic differences. Moreover, different measures of touchscreen learning were only modestly correlated in the BXD strains, indicating that these processes are comparatively independent at both genetic and phenotypic levels. Finally, we examined the behavioral structure of learning via principal component analysis of the current data, plus an archival dataset, totaling 765 mice. This revealed 5 independent factors suggestive of “reversal learning,” “motivation-related late reversal learning,” “discrimination learning,” “speed to respond,” and �

Relating high-temperature flow stress of AISI 316 stainless steel to strain and strain rate

International Nuclear Information System (INIS)

Matteazzi, S.; Paitti, G.; Boerman, D.

1982-01-01

The authors have performed an experimental determination of tensile stress-strain curves for different strain rates (4.67 x 10 - 5 , 4.67 x 10 - 2 s - 1 ) and for a variety of temperature conditions (773-1073 K) of AISI 316H stainless steel (annealed conditions) and also a computer analysis of the experimental curves using a fitting program which takes into consideration different constitutive relations describing the plastic flow behaviour of the metals. The results show that the materials tested are clearly affected by strain rate only at the highest temperature investigated (1073 K) and that the plastic strain is the more significant variable. Of the constitutive equations considered, Voce's relation gives the best fit for the true stress-time-strain curves. However, the Ludwik and Ludwigson equations also provide a description of the experimental data, whereas Hollomon's equation does not suitably characterize AISI 316H stainless steel and can be applied with some accuracy only at 1073 K. (author)

Evaluating location specific strain rates, temperatures, and accumulated strains in friction welds through microstructure modeling

Directory of Open Access Journals (Sweden)

Javed Akram

2018-04-01

Full Text Available A microstructural simulation method is adopted to predict the location specific strain rates, temperatures, grain evolution, and accumulated strains in the Inconel 718 friction welds. Cellular automata based 2D microstructure model was developed for Inconel 718 alloy using theoretical aspects of dynamic recrystallization. Flow curves were simulated and compared with experimental results using hot deformation parameter obtained from literature work. Using validated model, simulations were performed for friction welds of Inconel 718 alloy generated at three rotational speed i.e., 1200, 1500, and 1500 RPM. Results showed the increase in strain rates with increasing rotational speed. These simulated strain rates were found to match with the analytical results. Temperature difference of 150 K was noticed from center to edge of the weld. At all the rotational speeds, the temperature was identical implying steady state temperature (0.89Tm attainment. Keywords: Microstructure modeling, Dynamic recrystallization, Friction welding, Inconel 718, EBSD, Hot deformation, Strain map

Intraspecies diversity of Lactobacillus sakei response to oxidative stress and variability of strain performance in mixed strains challenges.

Science.gov (United States)

Guilbaud, Morgan; Zagorec, Monique; Chaillou, Stéphane; Champomier-Vergès, Marie-Christine

2012-04-01

Lactobacillus sakei is a meat-borne lactic acid bacterium species exhibiting a wide genomic diversity. We have investigated the diversity of response to various oxidative compounds, between L. sakei strains, among a collection representing the genomic diversity. We observed various responses to the different compounds as well as a diversity of response depending on the aeration conditions used for cell growth. A principal component analysis revealed two main phenotypic groups, partially correlating with previously described genomic clusters. We designed strains mixes composed of three different strains, in order to examine the behavior of each strain, when cultured alone or in the presence of other strains. The strains composing the mixtures were chosen as diverse as possible, i.e. exhibiting diverse responses to oxidative stress and belonging to different genomic clusters. Growth and survival rates of each strain were monitored under various aeration conditions, with or without heme supplementation. The results obtained suggest that some strains may act as "helper" or "burden" strains depending on the oxidative conditions encountered during incubation. This study confirms that resistance to oxidative stress is extremely variable within the L. sakei species and that this property should be considered when investigating starter performance in the complex meat bacterial ecosystems. Copyright © 2011 Elsevier Ltd. All rights reserved.

Genetic analysis of Saccharomyces cerevisiae strains isolated from palm wine in eastern Nigeria. Comparison with other African strains.

Science.gov (United States)

Ezeronye, O U; Legras, J-L

2009-05-01

To study the yeast diversity of Nigerian palm wines by comparison with other African strains. Twenty-three Saccharomyces cerevisiae strains were obtained from palm wine samples collected at four locations in eastern Nigeria, and characterized using different molecular techniques: internal transcribed spacer restriction fragment length polymorphism and sequence analysis, pulsed field gel electrophoresis, inter delta typing and microsatellite multilocus analysis. These techniques revealed that palm wine yeasts represent a group of closely related strains that includes other West African isolates (CBS400, NCYC110, DVPG6044). Population analysis revealed an excess of homozygote strains and an allelic richness similar to wine suggestive of local domestication. Several other African yeast strains were not connected to this group. Ghana sorghum beer strains and other African strains (DBVPG1853 and MUCL28071) displayed strikingly high relatedness with European bread, beer or wine strains, and the genome of strain MUCL30909 contained African and wine-type alleles, indicating its hybrid origin. Nigerian palm wine yeast represents a local specific yeast flora, whereas a European origin or hybrid was suspected for several other Africa isolates. This study presents the first genetic characterization of an autochthonous African palm wine yeast population and confirms the idea that human intervention has favoured yeast migration.

Internal residual strain mapping in carburized chrome molybdenum steel after quenching by neutron strain scanning

International Nuclear Information System (INIS)

Sakaida, Yoshihisa; Serizawa, Takanobu; Manzanka, Michiya

2011-01-01

A hollow circular cylinder specimen with an annular U-notch of chrome molybdenum steel with 0.20 mass% C (SCM420) was carburized in carrier gas and quenched in oil bath. In order to determine the case depth, the specimen was cut off and carbon content and Vickers hardness gradients were measured experimentally near the carburized surface. The residual strain mapping in the interior of carburized cylinder was conducted nondestructively by neutron strain scanning. In this study, the neutron diffraction from Fe-211 plane was used for strain scanning. The neutron wavelength was tuned to 0.1654nm so that diffraction angle became about 90deg. Radial, hoop and axial residual strains were measured by scanning diffracting volume along the axial direction of cylinder specimen. Each residual strain was calculated from lattice spacing change. Unstressed lattice spacing was determined experimentally using reference coupon specimens that were cut from the interior of same carburized cylinder. As a result, the diffraction peak width at half height, FWHM, near the carburized surface was about 3.7 times wider than that of coupon specimens. On the other hand, the most peak widths in the interior equaled to that of coupon specimens. Peak width broadened slightly as the diffracting volume approached the carburized case layer. From the center to the quarter of cylinder specimen, the hoop and axial strains were tensile, and the radial one was compressive in the interior. From the quarter to the edge of the cylinder specimen, the hoop tensile strain increased, radial and axial strains changed to tensile and compressive, respectively. Therefore, the interior of the cylinder specimen was found to be deformed elastically to balance the existence of compressive residual stresses in the carburized case layer. (author)

Strain gradient effects in surface roughening

DEFF Research Database (Denmark)

Borg, Ulrik; Fleck, N.A.

2007-01-01

evidence for strain gradient effects. Numerical analyses of a bicrystal undergoing in-plane tensile deformation are also studied using a strain gradient crystal plasticity theory and also by using a strain gradient plasticity theory for an isotropic solid. Both theories include an internal material length...

Nanocomposite Strain Gauges Having Small TCRs

Science.gov (United States)

Gregory, Otto; Chen, Ximing

2009-01-01

Ceramic strain gauges in which the strain-sensitive electrically conductive strips made from nanocomposites of noble metal and indium tin oxide (ITO) are being developed for use in gas turbine engines and other power-generation systems in which gas temperatures can exceed 1,500 F (about 816 C). In general, strain gauges exhibit spurious thermally induced components of response denoted apparent strain. When temperature varies, a strain-gauge material that has a nonzero temperature coefficient of resistance (TCR) exhibits an undesired change in electrical resistance that can be mistaken for the change in resistance caused by a change in strain. It would be desirable to formulate straingauge materials having TCRs as small as possible so as to minimize apparent strain. Most metals exhibit positive TCRs, while most semiconductors, including ITO, exhibit negative TCRs. The present development is based on the idea of using the negative TCR of ITO to counter the positive TCRs of noble metals and of obtaining the benefit of the ability of both ITO and noble metals to endure high temperatures. The noble metal used in this development thus far has been platinum. Combinatorial libraries of many ceramic strain gauges containing nanocomposites of various proportions of ITO and platinum were fabricated by reactive co-sputtering from ITO and platinum targets onto alumina- and zirconia-based substrates mounted at various positions between the targets.

Strain-dependent diffusion behavior of H within tungsten

International Nuclear Information System (INIS)

Ding, Wenyi; He, Haiyan; Liu, Changsong; Ding, Rui; Chen, Junling; Pan, Bicai

2014-01-01

The diffusion behaviors of H in tungsten, a promising material serving as the first wall facing the plasma in nuclear reactors, under either biaxial strain or isotropic strain are theoretically studied. We find that under the isotropic strain, an individual H atom may diffuse along all pathways, and under the biaxial strain, it preferably migrates along the direction perpendicular to the loaded strain. Moreover, in the case of either the isotropic or the biaxial strain, the loaded compressive strain weakens the diffusion of H, while the loaded tensile strain enhances the diffusion of H in bulk W.

Strain-dependent diffusion behavior of H within tungsten

Energy Technology Data Exchange (ETDEWEB)

Ding, Wenyi; He, Haiyan [Department of Physics, University of Science and Technology of China, Hefei, Anhui 230026 (China); Liu, Changsong [Key Laboratory of Materials Physics, Institute of Solid State Physics, Chinese Academy of Sciences, P.O. Box 1129, Hefei 230031 (China); Ding, Rui; Chen, Junling [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei 230031 (China); Pan, Bicai, E-mail: bcpan@ustc.edu.cn [Department of Physics, University of Science and Technology of China, Hefei, Anhui 230026 (China); Hefei National Laboratory for Physical Science at Microscale, University of Science and Technology of China, Hefei, Anhui 230026 (China)

2014-06-15

The diffusion behaviors of H in tungsten, a promising material serving as the first wall facing the plasma in nuclear reactors, under either biaxial strain or isotropic strain are theoretically studied. We find that under the isotropic strain, an individual H atom may diffuse along all pathways, and under the biaxial strain, it preferably migrates along the direction perpendicular to the loaded strain. Moreover, in the case of either the isotropic or the biaxial strain, the loaded compressive strain weakens the diffusion of H, while the loaded tensile strain enhances the diffusion of H in bulk W.

Use of colony-based bacterial strain typing for tracking the fate of Lactobacillus strains during human consumption

Directory of Open Access Journals (Sweden)

Drevinek Pavel

2009-12-01

Full Text Available Abstract Background The Lactic Acid Bacteria (LAB are important components of the healthy gut flora and have been used extensively as probiotics. Understanding the cultivable diversity of LAB before and after probiotic administration, and being able to track the fate of administered probiotic isolates during feeding are important parameters to consider in the design of clinical trials to assess probiotic efficacy. Several methods may be used to identify bacteria at the strain level, however, PCR-based methods such as Random Amplified Polymorphic DNA (RAPD are particularly suited to rapid analysis. We examined the cultivable diversity of LAB in the human gut before and after feeding with two Lactobacillus strains, and also tracked the fate of these two administered strains using a RAPD technique. Results A RAPD typing scheme was developed to genetically type LAB isolates from a wide range of species, and optimised for direct application to bacterial colony growth. A high-throughput strategy for fingerprinting the cultivable diversity of human faeces was developed and used to determine: (i the initial cultivable LAB strain diversity in the human gut, and (ii the fate of two Lactobacillus strains (Lactobacillus salivarius NCIMB 30211 and Lactobacillus acidophilus NCIMB 30156 contained within a capsule that was administered in a small-scale human feeding study. The L. salivarius strain was not cultivated from the faeces of any of the 12 volunteers prior to capsule administration, but appeared post-feeding in four. Strains matching the L. acidophilus NCIMB 30156 feeding strain were found in the faeces of three volunteers prior to consumption; after taking the Lactobacillus capsule, 10 of the 12 volunteers were culture positive for this strain. The appearance of both Lactobacillus strains during capsule consumption was statistically significant (p Conclusion We have shown that genetic strain typing of the cultivable human gut microbiota can be

Strain and strain rate by two-dimensional speckle tracking echocardiography in a maned wolf

Directory of Open Access Journals (Sweden)

Matheus M. Mantovani

2012-12-01

Full Text Available The measurement of cardiovascular features of wild animals is important, as is the measurement in pets, for the assessment of myocardial function and the early detection of cardiac abnormalities, which could progress to heart failure. Speckle tracking echocardiography (2D STE is a new tool that has been used in veterinary medicine, which demonstrates several advantages, such as angle independence and the possibility to provide the early diagnosis of myocardial alterations. The aim of this study was to evaluate the left myocardial function in a maned wolf by 2D STE. Thus, the longitudinal, circumferential and radial strain and strain rate were obtained, as well as, the radial and longitudinal velocity and displacement values, from the right parasternal long axis four-chamber view, the left parasternal apical four chamber view and the parasternal short axis at the level of the papillary muscles. The results of the longitudinal variables were -13.52±7.88, -1.60±1.05, 4.34±2.52 and 3.86±3.04 for strain (%, strain rate (1/s, displacement (mm and velocity (cm/s, respectively. In addition, the radial and circumferential Strain and Strain rate were 24.39±14.23, 1.86±0.95 and -13.69±6.53, -1.01±0.48, respectively. Thus, the present study provides the first data regarding the use of this tool in maned wolves, allowing a more complete quantification of myocardial function in this species.

Engineering piezoresistivity using biaxially strained silicon

DEFF Research Database (Denmark)

Pedersen, Jesper Goor; Richter, Jacob; Brandbyge, Mads

2008-01-01

of the piezocoefficient on temperature and dopant density is altered qualitatively for strained silicon. In particular, we find that a vanishing temperature coefficient may result for silicon with grown-in biaxial tensile strain. These results suggest that strained silicon may be used to engineer the iezoresistivity...

Strain path dependency in metal plasticity

NARCIS (Netherlands)

Viatkina, E.M.; Brekelmans, W.A.M.; Geers, M.G.D.

2003-01-01

A change in strain path has a significant effect on the mechanical response of metals. Strain path change effects physically originate from a complex microstructure evolution. This paper deals with the contribution of cell structure evolution to the strain path change effect. The material with cells

Sensibility of different wheat varieties (strains) to Ar+ implantation

International Nuclear Information System (INIS)

Cui Huanhu; Jing Hua; Ma Aiping; Kang Xiuli; Yang Liping; Huang Mingjing; Ma Buzhou; Shanxi Academy of Agricultural Sciences, Taiyuan

2005-01-01

The sensibility of different wheat varieties (strains) to Ar + implantation was studied. The results showed that the survival rate of 21 wheat varieties (strains) at the dose of 6 x 10 16 Ar + /cm 2 could be divided into five groups: surplus sensitive varieties (strains), sensitive varieties (strains), transitional varieties (strains), obtuse varieties (strains) and surplus obtuse varieties (strains). The sensibility of wheat varieties (strains) to Ar + injection is high-moisture-fertility wheat varieties (strains) > medium-moisture-fertility wheat varieties (strains) > dry land wheat varieties (strains). The study has provided theoretical basis in induced mutation medial lethal dose of different wheat varieties (strains) to Ar + implantation. (authors)

drug resistant strains of Salmonella enterica

African Journals Online (AJOL)

Conclusions: The aqueous extract of Thonningia sanguinea can provide an alternative therapy for the treatment of salmonellosis, mainly for typhoid fever caused by MDR strains of S. Typhi.The extract also inhibits S.Hadar a MDR emerging strain in Ivory Coast. Keywords: Thonningia sanguinea; Salmonella, MDR strains, ...

Validation of perceptual strain index to evaluate the thermal strain in experimental hot conditions

Directory of Open Access Journals (Sweden)

Habibollah Dehghan

2015-01-01

Conclusions: The research findings showed when there is no access to other forms of methods to evaluate the heat stress, it can be used the PeSI in evaluating the strain because of its favorable correlation with the thermal strain.

Mechanical properties of biaxially strained poly(L-lactide) tubes: Strain rate and temperature dependence

DEFF Research Database (Denmark)

Løvdal, Alexandra Liv Vest; Andreasen, Jens Wenzel; Mikkelsen, Lars Pilgaard

2017-01-01

Poly(l-lactide) (PLLA) is a bioabsorbable polymer with high stiffness and strength compared to the other commercially available bioabsorbable polymers. The properties of PLLA can be improved by straining, causing deformation-mediated molecular orientation. PLLA tubes were biaxially strained above...

StrainSeeker: fast identification of bacterial strains from raw sequencing reads using user-provided guide trees.

Science.gov (United States)

Roosaare, Märt; Vaher, Mihkel; Kaplinski, Lauris; Möls, Märt; Andreson, Reidar; Lepamets, Maarja; Kõressaar, Triinu; Naaber, Paul; Kõljalg, Siiri; Remm, Maido

2017-01-01

Fast, accurate and high-throughput identification of bacterial isolates is in great demand. The present work was conducted to investigate the possibility of identifying isolates from unassembled next-generation sequencing reads using custom-made guide trees. A tool named StrainSeeker was developed that constructs a list of specific k -mers for each node of any given Newick-format tree and enables the identification of bacterial isolates in 1-2 min. It uses a novel algorithm, which analyses the observed and expected fractions of node-specific k -mers to test the presence of each node in the sample. This allows StrainSeeker to determine where the isolate branches off the guide tree and assign it to a clade whereas other tools assign each read to a reference genome. Using a dataset of 100 Escherichia coli isolates, we demonstrate that StrainSeeker can predict the clades of E. coli with 92% accuracy and correct tree branch assignment with 98% accuracy. Twenty-five thousand Illumina HiSeq reads are sufficient for identification of the strain. StrainSeeker is a software program that identifies bacterial isolates by assigning them to nodes or leaves of a custom-made guide tree. StrainSeeker's web interface and pre-computed guide trees are available at http://bioinfo.ut.ee/strainseeker. Source code is stored at GitHub: https://github.com/bioinfo-ut/StrainSeeker.

Modulation of virulence and antibiotic susceptibility of enteropathogenic Escherichia coli strains by Enterococcus faecium probiotic strain culture fractions.

Science.gov (United States)

Ditu, Lia-Mara; Chifiriuc, Mariana Carmen; Bezirtzoglou, Eugenia; Voltsi, Chrysa; Bleotu, Coralia; Pelinescu, Diana; Mihaescu, Grigore; Lazar, Veronica

2011-12-01

The increasing rate of antimicrobial resistance drastically reduced the efficiency of conventional antibiotics and led to the reconsideration of the interspecies interactions in influencing bacterial virulence and response to therapy. The aim of the study was the investigation of the influence of the soluble and cellular fractions of Enterococcus (E.) faecium CMGB16 probiotic culture on the virulence and antibiotic resistance markers expression in clinical enteropathogenic Escherichia (E.) coli strains. The 7 clinical enteropathogenic E. coli strains, one standard E. coli ATCC 25,922 and one Bacillus (B.) cereus strains were cultivated in nutrient broth, aerobically at 37 °C, for 24 h. The E. faecium CMGB16 probiotic strain was cultivated in anaerobic conditions, at 37 °C in MRS (Man Rogosa Sharpe) broth, and co-cultivated with two pathogenic strains (B. cereus and E. coli O28) culture fractions (supernatant, washed sediment and heat-inactivated culture) for 6 h, at 37 °C. After co-cultivation, the soluble and cellular fractions of the probiotic strain cultivated in the presence of two pathogenic strains were separated by centrifugation (6000 rpm, 10 min), heat-inactivated (15 min, 100 °C) and co-cultivated with the clinical enteropathogenic E. coli strains in McConkey broth, for 24 h, at 37 °C, in order to investigate the influence of the probiotic fractions on the adherence capacity and antibiotic susceptibility. All tested probiotic combinations influenced the adherence pattern of E. coli tested strains. The enteropathogenic E. coli strains susceptibility to aminoglycosides, beta-lactams and quinolones was increased by all probiotic combinations and decreased for amoxicillin-clavulanic acid. This study demonstrates that the plurifactorial anti-infective action of probiotics is also due to the modulation of virulence factors and antibiotic susceptibility expression in E. coli pathogenic strains. Copyright © 2011 Elsevier Ltd. All rights reserved.

Strain distributions and their influence on electronic structures of WSe2-MoS2 laterally strained heterojunctions

Science.gov (United States)

Zhang, Chendong; Li, Ming-Yang; Tersoff, Jerry; Han, Yimo; Su, Yushan; Li, Lain-Jong; Muller, David A.; Shih, Chih-Kang

2018-02-01

Monolayer transition metal dichalcogenide heterojunctions, including vertical and lateral p-n junctions, have attracted considerable attention due to their potential applications in electronics and optoelectronics. Lattice-misfit strain in atomically abrupt lateral heterojunctions, such as WSe2-MoS2, offers a new band-engineering strategy for tailoring their electronic properties. However, this approach requires an understanding of the strain distribution and its effect on band alignment. Here, we study a WSe2-MoS2 lateral heterojunction using scanning tunnelling microscopy and image its moiré pattern to map the full two-dimensional strain tensor with high spatial resolution. Using scanning tunnelling spectroscopy, we measure both the strain and the band alignment of the WSe2-MoS2 lateral heterojunction. We find that the misfit strain induces type II to type I band alignment transformation. Scanning transmission electron microscopy reveals the dislocations at the interface that partially relieve the strain. Finally, we observe a distinctive electronic structure at the interface due to hetero-bonding.

Material Properties Test to Determine Ultimate Strain and True Stress-True Strain Curves for High Yield Steels

Energy Technology Data Exchange (ETDEWEB)

K.R. Arpin; T.F. Trimble

2003-04-01

This testing was undertaken to develop material true stress-true strain curves for elastic-plastic material behavior for use in performing transient analysis. Based on the conclusions of this test, the true stress-true strain curves derived herein are valid for use in elastic-plastic finite element analysis for structures fabricated from these materials. In addition, for the materials tested herein, the ultimate strain values are greater than those values cited as the limits for the elastic-plastic strain acceptance criteria for transient analysis.

Chemical Profile of Monascus ruber Strains

Directory of Open Access Journals (Sweden)

Ahamed M. Moharram

2012-01-01

Full Text Available Chemical profile of Monascus ruber strains has been studied using gas chromatography-mass spectrometry (GC/MS analysis. The colour intensity of the red pigment and secondary metabolic products of two M. ruber strains (AUMC 4066 and AUMC 5705 cultivated on ten different media were also studied. Metabolic products can be classified into four categories: anticholesterol, anticancer, food colouring, and essential fatty acids necessary for human health. Using GC/MS, the following 88 metabolic products were detected: butyric acid and its derivatives (25 products, other fatty acids and their derivatives (19 products, pyran and its derivatives (22 products and other metabolites (22 products. Among these, 32 metabolites were specific for AUMC 4066 strain and 34 for AUMC 5705 strain, whereas 22 metabolites were produced by both strains on different tested substrates. Production of some metabolites depended on the substrate used. High number of metabolites was recorded in the red pigment extract obtained by both strains grown on malt broth and malt agar. Also, 42 aroma compounds were recorded (4 alcohols, 2 benzaldehydes, 27 esters, 3 lactones, 1 phenol, 1 terpenoid, 3 thiol compounds and acetate-3-mercapto butyric acid. Thin layer chromatography and GC/MS analyses revealed no mycotoxin citrinin in any media used for the growth of the two M. ruber strains.

Effects of C and Si on strain aging of strain-based API X60 pipeline steels

Science.gov (United States)

Sung, Hyo Kyung; Lee, Dong Ho; Lee, Sunghak; Lee, Byeong-Joo; Hong, Seung-Pyo; Kim, Young-Woon; Yoo, Jang Yong; Hwang, Byoungchul; Shin, Sang Yong

2017-05-01

Four types of strain-based API X60 pipeline steels were fabricated by varying the C and Si contents, and the effects of C and Si on strain aging were investigated. The 0.05 wt% C steels consisted mainly of polygonal ferrite (PF), whereas the 0.08 wt% C steels consisted of acicular ferrite (AF). The volume fraction of AF increased with increasing C content because C is an austenite stabilizer element. The volume fractions of bainitic ferrite (BF) of the 0.15 wt% Si steels were higher than those of the 0.25 wt% Si steels, whereas the volume fractions of the secondary phases were lower. From the tensile properties before and after the aging process of the strainbased API X60 pipeline steels, the yield strength increased and the uniform and total elongation decreased, which is the strain aging effect. The strain aging effect in the strain-based API X60 pipeline steels was minimized when the volume fraction of AF was increased and secondary phases were distributed uniformly. On the other hand, an excessively high C content formed fine precipitates, and the strain aging effect occurred because of the interactions among dislocations and fine precipitates.

Evaluation of strain-rate sensitivity of ion-irradiated austenitic steel using strain-rate jump nanoindentation tests

Energy Technology Data Exchange (ETDEWEB)

Kasada, Ryuta, E-mail: r-kasada@iae.kyoto-u.ac.jp [Institute of Advanced Energy, Kyoto University Gokasho, Uji 611-0011, Kyoto (Japan); Konishi, Satoshi [Institute of Advanced Energy, Kyoto University Gokasho, Uji 611-0011, Kyoto (Japan); Hamaguchi, Dai; Ando, Masami; Tanigawa, Hiroyasu [Japan Atomic Energy Agency, Rokkasho, Aomori (Japan)

2016-11-01

Highlights: • We examined strain-rate jump nanoindentation on ion-irradiated stainless steel. • We observed irradiation hardening of the ion-irradiated stainless steel. • We found that strain-rate sensitivity parameter was slightly decreased after the ion-irradiation. - Abstract: The present study investigated strain-rate sensitivity (SRS) of a single crystal Fe–15Cr–20Ni austenitic steel before and after 10.5 MeV Fe{sup 3+} ion-irradiation up to 10 dpa at 300 °C using a strain-rate jump (SRJ) nanoindentation test. It was found that the SRJ nanoindentation test is suitable for evaluating the SRS at strain-rates from 0.001 to 0.2 s{sup −1}. Indentation size effect was observed for depth dependence of nanoindentation hardness but not the SRS. The ion-irradiation increased the hardness at the shallow depth region but decreased the SRS slightly.

5-Fluorouracil-resistant strain of Methanobacterium thermoautortrophicum

International Nuclear Information System (INIS)

Nagle, D.P. Jr.; Teal, R.; Eisenbraun, A.

1987-01-01

Growth of Methanobacterium thermoautotrophicum Marburg is inhibited by the pyrimidine, 5-fluorouracil (FU). It was shown previously that methanogenesis is not inhibited to the same extent as growth. A spontaneously occurring FU-resistant strain (RTAE-1) was isolated from a culture of strain Marburg. The growth of both strains was inhibited by 5-fluorodeoxyuridine but not 5-fluorocytosine, and the wild type was more susceptible to inhibition by 5-azauracil and 6-azauracil than was strain RTAE-1. The cellular targets for the pyrimidine analogs are not known. When the accumulation of 14 C-labeled uracil or FU by the two strains was compared, the wilt type took up 15-fold more radiolabel per cell than did the FU-resistant strain. In the wild type, radiolabel from uracil was incorporated into the soluble pool, RNA, and DNA. The metabolism of uracil appeared to involve a uracil phosphoribosyltransferase activity. Strain Marburg extracts contained this enzyme, whereas FU-resistant strain RTAE-1 extracts had less than 1/10 as much activity. Although it is possible that a change in permeability to the compounds plays a role in the stable resistance of strain RTAE-1, the fact that it lacks the ability to metabolize pyrimidines to nucleotides is sufficient to account for its phenotype

5-Fluorouracil-resistant strain of Methanobacterium thermoautotrophicum.

Science.gov (United States)

Nagle, D P; Teal, R; Eisenbraun, A

1987-09-01

Growth of Methanobacterium thermoautotrophicum Marburg is inhibited by the pyrimidine, 5-fluorouracil (FU). It was shown previously that methanogenesis is not inhibited to the same extent as growth. A spontaneously occurring FU-resistant strain (RTAE-1) was isolated from a culture of strain Marburg. The growth of both strains was inhibited by 5-fluorodeoxyuridine but not 5-fluorocytosine, and the wild type was more susceptible to inhibition by 5-azauracil and 6-azauracil than was strain RTAE-1. The cellular targets for the pyrimidine analogs are not known. When the accumulation of 14C-labeled uracil or FU by the two strains was compared, the wild type took up 15-fold more radiolabel per cell than did the FU-resistant strain. In the wild type, radiolabel from uracil was incorporated into the soluble pool, RNA, and DNA. The metabolism of uracil appeared to involve a uracil phosphoribosyltransferase activity. Strain Marburg extracts contained this enzyme, whereas FU-resistant strain RTAE-1 extracts had less than 1/10 as much activity. Although it is possible that a change in permeability to the compounds plays a role in the stable resistance of strain RTAE-1, the fact that it lacks the ability to metabolize pyrimidines to nucleotides is sufficient to account for its phenotype.

Complete Genomic Sequences of H3N8 Equine Influenza Virus Strains Used as Vaccine Strains in Japan.

Science.gov (United States)

Nemoto, Manabu; Yamanaka, Takashi; Bannai, Hiroshi; Tsujimura, Koji; Kokado, Hiroshi

2018-03-22

We sequenced the eight segments of influenza A virus strains A/equine/Ibaraki/1/2007 and A/equine/Yokohama/aq13/2010, which are strains of the Florida sublineage clades 1 and 2 of the H3N8 subtype equine influenza virus. These strains have been used as vaccine strains in Japan since 2016 in accordance with World Organization for Animal Health (OIE) recommendations. Copyright © 2018 Nemoto et al.

Strain accumulation in quasicrystalline solids

International Nuclear Information System (INIS)

Nori, F.; Ronchetti, M.; Elser, V.

1988-01-01

We study the relaxation of 2D quasicrystalline elastic networks when their constituent bonds are perturbed homogeneously. Whereas ideal, quasiperiodic networks are stable against such perturbations, we find significant accumulations of strain in a class of disordered networks generated by a growth process. The grown networks are characterized by root mean square phason fluctuations which grow linearly with system size. The strain accumulation we observe in these networks also grows linearly with system size. Finally, we find a dependence of strain accumulation on cooling rate

Towards development of nanofibrous large strain flexible strain sensors with programmable shape memory properties

Science.gov (United States)

Khalili, N.; Asif, H.; Naguib, H. E.

2018-05-01

Electrospun polymeric fibers can be used as strain sensors due to their large surface to weight/volume ratio, high porosity and pore interconnectivity. Large strain flexible strain sensors are used in numerous applications including rehabilitation, health monitoring, and sports performance monitoring where large strain detection should be accommodated by the sensor. This has boosted the demand for a stretchable, flexible and highly sensitive sensor able to detect a wide range of mechanically induced deformations. Herein, a physically cross-linked polylactic acid (PLA) and thermoplastic polyurethane (TPU) blend is made into nanofiber networks via electrospinning. The PLA/TPU weight ratio is optimized to obtain a maximum attainable strain of 100% while maintaining its mechanical integrity. The TPU/PLA fibers also allowed for their thermally activated recovery due to shape memory properties of the substrate. This novel feature enhances the sensor’s performance as it is no longer limited by its plastic deformation. Using spray coating method, a homogeneous layer of single-walled carbon nanotube is deposited onto the as-spun fiber mat to induce electrical conductivity to the surface of the fibers. It is shown that stretching and bending the sensor result in a highly sensitive and linear response with a maximum gauge factor of 33.

A closer look at prion strains

Science.gov (United States)

Solforosi, Laura; Milani, Michela; Mancini, Nicasio; Clementi, Massimo; Burioni, Roberto

2013-01-01

Prions are infectious proteins that are responsible for transmissible spongiform encephalopathies (TSEs) and consist primarily of scrapie prion protein (PrPSc), a pathogenic isoform of the host-encoded cellular prion protein (PrPC). The absence of nucleic acids as essential components of the infectious prions is the most striking feature associated to these diseases. Additionally, different prion strains have been isolated from animal diseases despite the lack of DNA or RNA molecules. Mounting evidence suggests that prion-strain-specific features segregate with different PrPSc conformational and aggregation states. Strains are of practical relevance in prion diseases as they can drastically differ in many aspects, such as incubation period, PrPSc biochemical profile (e.g., electrophoretic mobility and glycoform ratio) and distribution of brain lesions. Importantly, such different features are maintained after inoculation of a prion strain into genetically identical hosts and are relatively stable across serial passages. This review focuses on the characterization of prion strains and on the wide range of important implications that the study of prion strains involves. PMID:23357828

Local strains in waste tank deflagration analysis

International Nuclear Information System (INIS)

Bryan, B.J.; Flanders, H.E. Jr.

1993-01-01

In recent years extensive effort has been expended to qualify buried nuclear waste storage tanks under accident conditions. One of these conditions is deflagration of the combustible gases which may build up over time. While much work has been done to calculate the general strain state, less effort has been made to address the local strains at structural discontinuities. An analytical method is presented for calculating these local strains and combining them with the general strain state. A closed form solution of the local strains is compared to a finite element solution

Spontaneous abortion and physical strain around implantation

DEFF Research Database (Denmark)

Hjøllund, Niels Henrik Ingvar; Jensen, T.K.; Bonde, J.P.

2000-01-01

Existing studies of physical strain and spontaneous abortion are mainly retrospective or based only on pregnancies that have survived the first trimester. Furthermore, almost all studies have relied on averaged measures of physical strain, which tend to blur an effect if peak values during short...... pregnancy the women recorded physical strain prospectively in a structured diary. Physical strain around the time of implantation was associated with later spontaneous abortion. The adjusted risk ratio for women who reported physical strain higher than average at day 6 to 9 after the estimated date...

Strain characterization of FinFETs using Raman spectroscopy

International Nuclear Information System (INIS)

Kaleli, B.; Hemert, T. van; Hueting, R.J.E.; Wolters, R.A.M.

2013-01-01

Metal induced strain in the channel region of silicon (Si) fin-field effect transistor (FinFET) devices has been characterized using Raman spectroscopy. The strain originates from the difference in thermal expansion coefficient of Si and titanium-nitride. The Raman map of the device region is used to determine strain in the channel after preparing the device with the focused ion beam milling. Using the Raman peak shift relative to that of relaxed Si, compressive strain values up to – 0.88% have been obtained for a 5 nm wide silicon fin. The strain is found to increase with reducing fin width though it scales less than previously reported results from holographic interferometry. In addition, finite-element method (FEM) simulations have been utilized to analyze the amount of strain generated after thermal processing. It is shown that obtained FEM simulated strain values are in good agreement with the calculated strain values obtained from Raman spectroscopy. - Highlights: ► Strain is characterized in nanoscale devices with Raman spectroscopy. ► There is a fin width dependence of the originated strain. ► Strain levels obtained from this technique is in correlation with device simulations

Genetic diversity of citrus bacterial canker pathogens preserved in herbarium specimens.

Science.gov (United States)

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

2007-11-20

Citrus bacterial canker (CBC) caused by Xanthomonas axonopodis pv. citri (Xac) was first documented in India and Java in the mid 19th century. Since that time, the known distribution of the disease has steadily increased. Concurrent with the dispersion of the pathogen, the diversity of described strains continues to increase, with novel strains appearing in Saudi Arabia, Iran, and Florida in the last decade. Herbarium specimens of infected plants provide an historical record documenting both the geographic distribution and genetic diversity of the pathogen in the past. However, no method was available to assess the genetic diversity within these herbarium samples. We have developed a method, insertion event scanning (IES), and applied the method to characterize the diversity present within CBC populations documented as herbarium specimens over the past century. IES is based on the specific amplification of junction fragments that define insertion events. The potential for IES in current forensic applications is demonstrated by finding an exact match of pathogen genotypes preserved in herbarium specimens from Japan and Florida, demonstrating the source of the original outbreak of citrus canker in Florida in 1911. IES is a very sensitive technique for differentiating bacterial strains and can be applied to any of the several hundred bacteria for which full genomic sequence data are available.

An Ecoinformatics Approach to Field-Scale Evaluation of Insecticide Effects in California Citrus: Are Citrus Thrips and Citrus Red Mite Induced Pests?

Science.gov (United States)

Livingston, George; Hack, Lindsey; Steinmann, Kimberly P; Grafton-Cardwell, Elizabeth E; Rosenheim, Jay A

2018-05-28

Experimental approaches to studying the consequences of pesticide use, including impacts on beneficial insects, are vital; however, they can be limited in scale and realism. We show that an ecoinformatics approach that leverages existing data on pesticides, pests, and beneficials across multiple fields can provide complementary insights. We do this using a multi-year dataset (2002-2013) on pesticide applications and density estimates of two pests, citrus thrips (Scirtothrips citri (Moulton [Thysanoptera: Thripidae])) and citrus red mites (Panonychus citri McGregor [Acari: Tetranychidae]), and a natural enemy (Euseius spp. predatory mites) collected from citrus groves in the San Joaquin Valley of California. Using correlative analyses, we investigated the long-term consequences of pesticide use on S. citri and P. citri population densities to evaluate the hypothesis that the pest status of these species is largely due to the disruption of natural biological control-i.e., these are induced pests. We also evaluated short-term pesticide efficacy (suppression of citrus thrips and citrus red mite populations immediately post-application) and asked if it was correlated with the suppression of Euseius predator populations. Although the short-term efficacy of different pesticides varied significantly, our dataset does not suggest that the use of citrus pesticides suppressed Euseius densities or worsened pest problems. We also find that there is no general trade-off between pesticide efficacy and pesticide risk to Eusieus, such that highly effective and minimally disruptive compounds were available to citrus growers during the studied time period.

Strain bidimensional na cardiopatia de Takotsubo Two-dimensional strain in Takotsubo cardiomyopathy

Directory of Open Access Journals (Sweden)

Carlos Bellini G. Gomes

2010-08-01

Full Text Available Este relato apresenta o seguimento tardio de um caso de cardiomiopatia de Takotsubo com boa evolução clínica e melhora da função sistólica global ventricular esquerda. Contudo, observou-se persistência de significativa disfunção sistólica regional longitudinal que foi avaliada por meio de nova técnica ecocardiográfica (speckle tracking, com as medidas do strain (S e strain rate (SR correspondentes. Ressaltamos a importância desse novo método para o acompanhamento dessa cardiopatia, pois permite identificar os pacientes que persistem com disfunção sistólica e que possivelmente serão beneficiados com a manutenção da terapêutica clínica.This report presents the late follow-up of a case of Takotsubo cardiomyopathy with good clinical outcome and improved left ventricular global systolic function. However, there was persistence of significant regional longitudinal systolic dysfunction evaluated using a new echocardiographic technique (speckle tracking, with corresponding measures of strain (S and strain rate (SR. We emphasize the importance of this new method to monitoring this cardiomyopathy, since it identifies patients with persistent systolic dysfunction who will possibly benefit from maintenance of clinical therapy

Resonant tunneling measurements of size-induced strain relaxation

Science.gov (United States)

Akyuz, Can Deniz

Lattice mismatch strain available in such semiconductor heterostructures as Si/SiGe or GaAs/AlGaAs can be employed to alter the electronic and optoelectronic properties of semiconductor structures and devices. When deep submicron structures are fabricated from strained material, strained layers relax by sidewall expansion giving rise to size- and geometry-dependent strain gradients throughout the structure. This thesis describes a novel experimental technique to probe the size-induced strain relaxation by studying the tunneling current characteristics of strained p-type Si/SiGe resonant tunneling diodes. Our current-voltage measurements on submicron strained p-Si/SiGe double- and triple-barrier resonant tunneling structures as a function of device diameter, D, provide experimental access to both the average strain relaxation (which leads to relative shifts in the tunneling current peak positions) and strain gradients (which give rise to a fine structure in the current peaks due to inhomogeneous strain-induced lateral quantization). We find that strain relaxation is significant, with a large fraction of the strain energy relaxed on average in D ≤ 0.25 m m devices. Further, the in-plane potentials that arise from inhomogeneous strain gradients are large. In the D ˜ 0.2 m m devices, the corresponding lateral potentials are approximately parabolic exceeding ˜ 25 meV near the perimeter. These potentials create discrete hole states in double-barrier structures (single well), and coupled hole states in triple-barrier structures (two wells). Our results are in excellent agreement with finite-element strain calculations in which the strained layers are permitted to relax to a state of minimum energy by sidewall expansion. Size-induced strain relaxation will undoubtedly become a serious technological issue once strained devices are scaled down to the deep submicron regime. Interestingly, our calculations predict and our measurements are consistent with the appearance of

Strain Rate Effect on Tensile Behavior for a High Specific Strength Steel: From Quasi-Static to Intermediate Strain Rates

Directory of Open Access Journals (Sweden)

Wei Wang

2017-12-01

Full Text Available The strain rate effect on the tensile behaviors of a high specific strength steel (HSSS with dual-phase microstructure has been investigated. The yield strength, the ultimate strength and the tensile toughness were all observed to increase with increasing strain rates at the range of 0.0006 to 56/s, rendering this HSSS as an excellent candidate for an energy absorber in the automobile industry, since vehicle crushing often happens at intermediate strain rates. Back stress hardening has been found to play an important role for this HSSS due to load transfer and strain partitioning between two phases, and a higher strain rate could cause even higher strain partitioning in the softer austenite grains, delaying the deformation instability. Deformation twins are observed in the austenite grains at all strain rates to facilitate the uniform tensile deformation. The B2 phase (FeAl intermetallic compound is less deformable at higher strain rates, resulting in easier brittle fracture in B2 particles, smaller dimple size and a higher density of phase interfaces in final fracture surfaces. Thus, more energy need be consumed during the final fracture for the experiments conducted at higher strain rates, resulting in better tensile toughness.

Stress and strain measurements in solids

International Nuclear Information System (INIS)

Askegaard, V.

1978-01-01

A design basis is given for stress- and strain cells to be used in a solid either externally loaded or with a stressfree strain field (for example shrinkage). A stress- and a strain cell has been designed for use in granular materials. Calibration tests show either good or reasonably good correspondance with calculated values. (orig.) [de

Strain specific variation of outer membrane proteins of wild Yersinia pestis strains subjected to different growth temperatures

Directory of Open Access Journals (Sweden)

Frederico Guilherme Coutinho Abath

1990-03-01

Full Text Available Three Yersinia pestis strains isolated from humans and one laboratory strain (EV76 were grown in rich media at 28§C and 37§C and their outer membrane protein composition compared by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-Page. Several proteins with molecular weights ranging from 34 kDa to 7 kDa were observed to change in relative abundance in samples grown at different temperatures. At least seven Y. pestis outer membrane proteins showed a temperature-dependent and strain-specific behaviour. Some differences between the outer membrane proteins of full-pathogenic wild isolates and the EV76 strain could aldso be detected and the relevance of this finding on the use of laboratory strains as a reference to the study of Y. pestis biological properties is discuted.

Properties of strain gages at cryogenic temperature

International Nuclear Information System (INIS)

Shibata, Nobuo; Fujiyoshi, Toshimitsu.

1978-01-01

At the time of developing superconduction generators, the stress measurement for rotor parts is required to grasp the safety and performance of the rotor at cryogenic temperature, which is cooled with liquid helium. In case of carrying out the stress measurement with strain gages, the problems are as follows. The strain gages and lead wires are exposed to cryogenic temperature from 4 to 10 K and strong magnetic field of about 3T, and subjected to high centrifugal acceleration of about 500G. In order to establish the techniques of the stress measurement under such conditions, the adhesives and damp-proof coatings for strain gages and strain gages themselves in Japan and foreign countries were examined on the properties at cryogenic temperature. As for the properties of strain gages, mainly the apparent strain owing to temperature change was investigated, and the change of the gage factors was studies only at liquid nitrogen temperature. The stress measurement with strain gages at low temperature had been studied in detail down to liquid nitrogen temperature concerning LNG tanks. The experimental apparatus, the samples, the testing methods and the test results of cooling tests on adhesives and damp-proof coatings, and the temperature characteristics of strain gages are reported. The usable adhesives and coatings were found, and correction by accurate temperature measurement is required for apparent strain. (Kako, I.)

Resolution of axial shear strain elastography

International Nuclear Information System (INIS)

Thitaikumar, Arun; Righetti, Raffaella; Krouskop, Thomas A; Ophir, Jonathan

2006-01-01

The technique of mapping the local axial component of the shear strain due to quasi-static axial compression is defined as axial shear strain elastography. In this paper, the spatial resolution of axial shear strain elastography is investigated through simulations, using an elastically stiff cylindrical lesion embedded in a homogeneously softer background. Resolution was defined as the smallest size of the inclusion for which the strain value at the inclusion/background interface was greater than the average of the axial shear strain values at the interface and inside the inclusion. The resolution was measured from the axial shear strain profile oriented at 45 0 to the axis of beam propagation, due to the absence of axial shear strain along the normal directions. The effects of the ultrasound system parameters such as bandwidth, beamwidth and transducer element pitch along with signal processing parameters such as correlation window length (W) and axial shift (ΔW) on the estimated resolution were investigated. The results show that the resolution (at 45 0 orientation) is determined by the bandwidth and the beamwidth. However, the upper bound on the resolution is limited by the larger of the beamwidth and the window length, which is scaled inversely to the bandwidth. The results also show that the resolution is proportional to the pitch and not significantly affected by the axial window shift

Influence of dynamic strain ageing on tensile strain energy of type 316L(N) austenitic stainless steel

International Nuclear Information System (INIS)