San Joaquin, CA, United States
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Nwugo C.C.,San Joaquin Valley Agricultural science Center | Duan Y.,U.S. Department of Agriculture | Lin H.,San Joaquin Valley Agricultural science Center
PLoS ONE | Year: 2013

Citrus huanglongbing (HLB) is a highly destructive disease of citrus presumably caused by 'Candidatus Liberibacter asiaticus' (Las), a gram-negative, insect-transmitted, phloem-limited α-proteobacterium. Although almost all citrus plants are susceptible to HLB, reports have shown reduced susceptibility to Las infection in lemon (Citrus limon) plants. The aim of this study is to identify intra-species specific molecular mechanisms associated with Las-induced responses in lemon plants. To achieve this, comparative 2-DE and mass spectrometry, in addition to Inductively Coupled Plasma Spectroscopy (ICPS) analyses, were applied to investigate differences in protein accumulation and the concentrations of cationic elements in leaves of healthy and Las-infected lemon plants. Results showed a differential accumulation of 27 proteins, including an increase in accumulation of starch synthase but decrease in the production of photosynthesis-related proteins in Las-infected lemon plants compared to healthy plants. Furthermore, there was a 6% increase (P > 0.05) in K concentration in leaves of lemon plants upon Las infection, which support results from previous studies and might represent a common response pattern of citrus plants to Las infection. Interestingly, contrary to reports from prior studies, this study showed a general reduction in the production of defense-related pathogen-response proteins but a 128% increase in Zn concentration in lemon plants in response to Las infection. Taken together, this study sheds light on general and intra-species specific responses associated with the response of citrus plants to Las. © 2013 Nwugo et al.


Bellamy D.E.,San Joaquin Valley Agricultural science Center | Sisterson M.S.,San Joaquin Valley Agricultural science Center | Walse S.S.,San Joaquin Valley Agricultural science Center
PLoS ONE | Year: 2013

Novel methodology is presented for indexing the relative potential of hosts to function as resources. A Host Potential Index (HPI) was developed as a practical framework to express relative host potential based on combining results from one or more independent studies, such as those examining host selection, utilization, and physiological development of the organism resourcing the host. Several aspects of the HPI are addressed including: 1) model derivation; 2) influence of experimental design on establishing host rankings for a study type (no choice, two-choice, and multiple-choice); and, 3) variable selection and weighting associated with combining multiple studies. To demonstrate application of the HPI, results from the interactions of spotted wing drosophila (SWD), Drosophila suzukii Matsumura (Diptera: Drosophilidae), with seven "reported" hosts (blackberries, blueberries, sweet cherries, table grapes, peaches, raspberries, and strawberries) in a postharvest scenario were analyzed. Four aspects of SWD-host interaction were examined: attraction to host volatiles; population-level oviposition performance; individual-level oviposition performance; and key developmental factors. Application of HPI methodology indicated that raspberries (meanHPIvaried = 301.9±8.39; rank 1 of 7) have the greatest potential to serve as a postharvest host for SWD relative to the other fruit hosts, with grapes (meanHPIvaried = 232.4±3.21; rank 7 of 7) having the least potential.


Backus E.A.,San Joaquin Valley Agricultural science Center | Morgan D.J.W.,Mt. Rubidoux Field Station
Phytopathology | Year: 2011

The pathogen that causes Pierce's disease of grapevine, Xylella fastidiosa, is the only known bacterial, arthropod-transmitted plant pathogen that does not circulate in the vector's hemolymph. Instead, bacteria are foregut-borne, persistent in adult vectors but semipersistent in immatures (i.e., bacteria colonize cuticular surfaces of the anterior foregut, are retained for hours to days, but are lost during molting). Yet, exactly how a sharpshooter vector inoculates bacteria from foregut acquisition sites is unknown. The present study used confocal laserscanning microscopy to identify locations in undissected, anterior foreguts of the glassy-winged sharpshooter colonized by green fluorescent protein-expressing X. fastidiosa. Spatial and temporal distributions of colonizing X. fastidiosa were examined daily over acquisition access periods of 1 to 6 days for both contaminated field-collected and clean laboratory-reared Homalodisca vitripennis. Results provide the first direct, empirical evidence that established populations of X. fastidiosa can disappear from vector foreguts over time. When combined with existing knowledge on behavior, physiology, and functional anatomy of sharpshooter feeding, present results support the idea that the disappearance is caused by outward fluid flow (egestion) not inward flow (ingestion) (i.e., swallowing). Thus, results support the hypothesis that egestion is a critical part of the X. fastidiosa inoculation mechanism. Furthermore, results suggest a cyclical, spatiotemporal pattern of microbial colonization, disappearance, and recolonization in the precibarium. Colonization patterns also support two types of egestion, termed rinsing and discharging egestion herein. Finally, comparison of acquisition results for field-collected versus laboratory-reared sharpshooters suggest that there may be competitive binding for optimum acquisition sites in the foregut. Therefore, successful inoculation of X. fastidiosa may depend, in large part, on vector load in the precibarium. © 2011 The American Phytopathological Society.


Kuenen L.P.S.,San Joaquin Valley Agricultural science Center | Siegel J.P.,San Joaquin Valley Agricultural science Center
Environmental Entomology | Year: 2010

The navel orangeworm, Amyelois transitella (Walker), is the primary insect pest of pistachios and almonds in California. Four years of research (20022006) were conducted in Madera and Kern Counties to elucidate the pattern of adult emergence of the overwintering navel orangeworm population. Springtime emergence from unharvested (mummy) nuts was protracted (600 degree-days or more from 1 January of each year) and in 2004 and 2006 extended to mid-July. The population structure, sex ratio, and timing of emergence differed between pistachio and almond mummies, Pistachio populations had a significantly greater proportion of late stage individuals compared with almond mummies, 85.7 versus 34.1%. The sex ratio of adults emerging from pistachio mummies was significantly skewed with a ratio 57:43 male:female compared with 50:50 in almond mummies. Emergence from mummies held outdoors (variable temperature) began in early March and continued through early June in both pistachio mummies and almond mummies. The adult emergence pattern from pistachio mummies contained a single emergence peak, whereas emergence from almond mummies occurred in multiple peaks. These same patterns occurred when mummies were held at constant temperature, and the emergence peak from pistachio mummies occurred sooner. The impact of these findings on understanding navel orangeworm population dynamics and current control recommendations is discussed. © 2010 Entomological Society of America.


Sisterson M.S.,San Joaquin Valley Agricultural science Center
Journal of Economic Entomology | Year: 2014

A method to improve an assay relating adult feeding to egg maturation by the glassywinged sharpshooter (Hemiptera: Cicadellidae) was evaluated. The assay consisted on confining females to cowpea stems and quantifying feeding and egg maturation. Feeding was quantified by measuring excreta production. The number of eggs matured during the assay was estimated by taking the difference between female egg load (number of mature eggs carried by a female) at end of the assay (determined by dissection) and mean egg load of a subset of females dissected at start of the assay. Estimates of the number of mature eggs produced by females using the aforementioned approach improve as variability in egg loads of females entering the assay declines. As egg loads of females are variable, a pretreatment designed to reduce variance in egg loads of females entering the assay was evaluated. To accomplish this, females were divided into two groups. The control group was placed directly into the assay. The pretreatment group was given an oviposition period on sorghum before the assay. An oviposition period on sorghum was expected to reduce variance in egg load among females, as previous research found that sorghum was suitable for oviposition but provided poor nutrition for egg maturation. Dissection of a subset of females from each group before the assay determined that the mean and variance in egg load of females receiving the pretreatment was significantly reduced compared with females in the control group. Analysis of results from the feeding assay found that there was a significant relationship between feeding and egg maturation for females receiving the pretreatment, but not for females in the control group. Thus, reducing the mean and variance in egg load of females entering feeding assays resulted in detection of a significant positive relationship between feeding and egg maturation that otherwise would not have been observed. © 2014 Entomological Society of America.


Yokoyama V.Y.,San Joaquin Valley Agricultural science Center
Environmental Entomology | Year: 2012

The mobility of olive fruit fly, Bactrocera oleae (Rossi), late third instars before pupation, teneral adults before flight, and mature adults restricted from flight were studied under mulches in greenhouse cage tests, in horizontal pipes, vertical bottles and pipes filled with sand, and by observation on smooth laboratory surfaces. Percentage adults emerging from pupae and percentage adult females that escaped soil, fabric, and paper mulches over a soil or sand substrate ranged from 63 to 83, and 4053%, respectively. Percentage adults emerging from pupae and percentage adult females that walked through the open interior of 1.526.10-m horizontal pipes of 1.52.0-cm inner diameter ranged from 57 to 81, and 2761%, respectively. Percentage adults emerging from pupae that escaped through sand depths of 2.510.2, and 12.720.3 cm, ranged from 68 to 87, and 1288%; and percentage adult females that escaped ranged from 46 to 58, and 3870%, respectively. In 15.4-cm-inner-diameter pipes filled with different heights of sand, the highest percentage of the total number of adults that emerged in the control were found from 0 to 20.3 cm, and ranged from 37 to 71%. Ten to 47% of adults were found from 20.3 cm to below the surface, and 621% escaped to the top of 20.350.8 cm high sand columns. In column heights of 55.9 and 61 cm, pressures at the bottom caused by the weight of the sand above were 91.4 and 99.7 g/cm2, respectively, and a mean of <1 adult escaped to the top. Before pupation, the late third instars were found to travel continuously for 6.9 h over 23.9 m at a speed of 6.0 cm per min, when placed on a smooth surface, at 22.2°C. Teneral females and males that could not fly, made ≈7 stops totaling 1113 min, walked at a speed of 5762 cm per min, and began a rest period of 8384 min duration, at 8589 min before flight. Males walked a distance of 13.1 m in 22 min, which was greater than females that walked for 9.6 m in 17 min, at 2022°C and 35% RH. The mobility of the third instars and the teneral adults is discussed in relation to potential control techniques in olive orchards. © 2012 Entomological Society of America.


Stenger D.C.,San Joaquin Valley Agricultural science Center | Lee M.W.,San Joaquin Valley Agricultural science Center
Applied and Environmental Microbiology | Year: 2011

Incompatibility group P1 (IncP-1) plasmid diversity was evaluated based on replication initiator protein (TrfA) phylogeny. A new and highly divergent clade was identified. Replication assays indicated that TrfA of recently discovered IncP-1 plasmids from Xylella fastidiosa and Verminephrobacter eiseniae initiated plasmid replication using cognate or heterologous origins of replication. © 2011, American Society for Microbiology.


Burbank L.P.,San Joaquin Valley Agricultural science Center | Stenger D.C.,San Joaquin Valley Agricultural science Center
Phytopathology | Year: 2016

The phytopathogen Xylella fastidiosa causes disease in a variety of important crop and landscape plants. Functional genetic studies have led to a broader understanding of virulence mechanisms used by this pathogen in the grapevine host. Plasmid shuttle vectors are important tools in studies of bacterial genetics but there are only a limited number of plasmid vectors available that replicate in X. fastidiosa, and even fewer that are retained without antibiotic selection. Two plasmids are described here that show stable replication in X. fastidiosa and are effective for gene complementation both in vitro and in planta. Plasmid maintenance is facilitated by incorporation of the PemI/PemK plasmid addiction system, consisting of PemK, an endoribonuclease toxin, and its cognate antitoxin, PemI. Vector pXf20pemIK utilizes a native X. fastidiosa replication origin as well as a high-copy-number pUC origin for propagation in Escherichia coli cloning strains. Broad-host-range vector pBBR5pemIK is a medium- to low-copy-number plasmid based on the pBBR1 backbone. Both plasmids are maintained for extended periods of time in the absence of antibiotic selection, as well as up to 14 weeks in grapevine, without affecting bacterial fitness. These plasmids present an alternative to traditional complementation and expression vectors which rely on antibiotic selection for plasmid retention. © 2016, American Phytopathological Society. All rights reserved.


Spear A.,San Joaquin Valley Agricultural science Center | Sisterson M.S.,San Joaquin Valley Agricultural science Center | Stenger D.C.,San Joaquin Valley Agricultural science Center
Virus Research | Year: 2012

A complex set of double-stranded RNAs (dsRNAs) was isolated from threecornered alfalfa hopper (Spissistilus festinus), a plant-feeding hemipteran pest. A subset of these dsRNAs constitute the genome of a new reovirus, provisionally designated Spissistilus festinus reovirus (SpFRV). SpFRV was present in threecornered alfalfa hopper populations in the San Joaquin Valley of California, with incidence ranging from 10% to 60% in 24 of 25 sample sets analyzed. The 10 dsRNA segments of SpFRV were completely sequenced and shown to share conserved terminal sequences (5'-AGAGA and CGAUGUUGU-3') of the positive-sense strand that are distinct from known species of the family Reoviridae. Comparisons of the RNA directed RNA polymerase (RdRp) indicated SpFRV is most closely related (39.1% amino acid identity) to another new reovirus infecting the angulate leafhopper (Acinopterus angulatus) and provisionally designated Acinopterus angulatus reovirus (AcARV). The RdRp of both viruses was distantly related to Raspberry latent virus RdRp at 27.0% (SpFRV) and 30.0% (AcARV) or Rice ragged stunt virus RdRp at 26.2% (SpFRV) and 29.0% (AcARV) amino acid identity. RdRp phylogeny confirmed that SpFRV and AcARV are sister taxa sharing a most recent common ancestor. SpFRV segment 6 encodes a protein containing two NTP binding motifs that are conserved in homologs of reoviruses in the subfamily Spinareovirinae. The protein encoded by SpFRV segment 4 was identified as a guanylyltransferase homolog. SpFRV segments 1, 3, and 10 encode homologs of reovirus structural proteins. No homologs were identified for proteins encoded by SpFRV segments 5, 7, 8, and 9. Collectively, the low level of sequence identity with other reoviruses, similar segment terminal sequences, RdRp phylogeny, and host taxa indicate that SpFRV and AcARV may be considered members of a proposed new genus of the family Reoviridae (subfamily Spinareovirinae), with SpFRV assigned as the type species. © 2011.


Wallis C.M.,San Joaquin Valley Agricultural science Center | Chen J.,San Joaquin Valley Agricultural science Center
Phytopathology | Year: 2012

Pierce's disease of grapevine (PD), caused by the bacterial pathogen Xylella fastidiosa, remains a serious problem for grape production in California and elsewhere. This research examined induction of phenolic compounds in grapevines ('Thompson Seedless') infected with X. fastidiosa over a 6-month period. Two months postinoculation with X. fastidiosa, catechin, digalloylquinic acid, and astringin were found at greater levels in xylem sap; multiple catechins, procyanidins, and stilbenoids were found at greater levels in xylem tissues; and precursors to lignin and condensed tannins were found at greater levels in xylem cell walls. However, such large-scale inductions of phenolic compounds were not observed 4 months after inoculation. Six months after inoculation, infected plants had significantly reduced phenolic levels in xylem sap and tissues when compared with control plants, including lowered levels of lignin and condensed tannins. At 6 months, PD symptoms were severe in infected plants and most photosynthetic tissue was abscised. These results suggest that, even though grapevine hosts may initially respond to X. fastidiosa infections with increased production of phenolic compounds, ultimately, PD causes grapevines to enter a state of decline whereby diseased hosts no longer have the resources to support secondary metabolite production, including defense-associated phenolic compounds.

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