Planococcus ficus (Hemiptera: Pseudococcidae) movement and demography: methods for generating cohorts for laboratory studies.

Planococcus ficus (Signoret) is a worldwide pest of grapevine. Mealybugs overwinter under bark and move into the grape canopy as the season progresses. Because crawlers are more active than later stages, mealybug movement behavior is likely to be stage specific. To quantify P. ficus demography and movement behavior, a series of laboratory experiments were conducted. First, P. ficus populations were monitored on grapevine seedlings to describe survival, change in size, timing of male pupation, and timing of oviposition over a 6-wk period. Subsequently, cohorts of mealybugs were generated by infesting grapevines with crawlers and holding infested grapevines for a specified duration of 0 (crawlers), 1, 2, 3, or 4 wk. Crawlers (0-wk) were more likely to move upwards and towards a light source, than all other age cohorts tested. Further, mealybugs from 4-wk-old cohorts were more likely to move downward than all other age cohorts tested. Results suggest that crawlers are more likely to move to the top of grapevines by moving upwards and orienting towards either the sun or the moon than all other age cohorts tested, whereas older gravid females are more likely to move downward. Passive movement of mealybugs on farm machinery or animals requires surviving a host free period. To quantify risk of passive movement, establishment rates and effects of starvation on each age cohort were quantified. Larger and older mealybugs were more likely to establish on grapevines than smaller and younger mealybugs. Further, mealybug longevity in absence of food was greater for older cohorts compared to younger cohorts. Crawlers survived an average of 2 days without food, whereas females from 4-wk-old cohorts survived for an average of 11 days without food. Further, 70% of starved females from 4-wk-old cohorts deposited fertile eggs. In the absence of food, some mealybugs from cohorts aged 2-, 3-, and 4-wk formed pupa with viable males emerging. Adult males from starved nymphs lived for an average of 3 days post-emergence. Results provide methods for producing cohorts of mealybugs of predictable size and stage and provides insight into P. ficus demography and movement behavior.

High Growing Season Temperatures Limit Winter Recovery of Grapevines from Xylella fastidiosa Infection - Implications for Epidemiology in Hot Climates.

Management of widespread plant pathogens is challenging as climatic differences among crop-growing regions may alter key aspects of pathogen spread and disease severity. Xylella fastidiosa is a xylem-limited bacterial pathogen that is transmitted by xylem sap-feeding insects. Geographic distribution of X. fastidiosa is limited by winter climate, and vines infected with X. fastidiosa can recover from infection when held at cold temperatures. California has a long history of research on Pierce's disease and significant geographic and climatic diversity among grape-growing regions. This background in combination with experimental disease studies under controlled temperature conditions can inform risk assessment for X. fastidiosa spread and epidemic severity across different regions and under changing climate conditions. California's grape-growing regions have considerable differences in summer and winter climate. In northern and coastal regions, summers are mild and winters are cool, conditions which favor winter recovery of infected vines. In contrast, in inland and southern areas, summers are hot and winters mild, reducing likelihood of winter recovery. Here, winter recovery of three table grape cultivars (Flame, Scarlet Royal, and Thompson Seedless) and three wine grape cultivars (Sauvignon Blanc, Cabernet Sauvignon, and Zinfandel) were evaluated under temperature conditions representative of the San Joaquin Valley, an area with hot summers and mild winters that has been severely impacted by Pierce's disease and contains a large portion of California grape production. Mechanically inoculated vines were held in the greenhouse under one of three warming treatments to represent different seasonal inoculation dates prior to being moved into a cold chamber. Winter recovery under all treatments was generally limited but with some cultivar variation. Given hot summer temperatures of many grape-growing regions worldwide, as well as increasing global temperatures overall, winter recovery of grapevines should not be considered a key factor limiting X. fastidiosa spread and epidemic severity in the majority of cases.

Glassy-Winged Sharpshooters Cease Feeding and Avoid Plants Treated With Sub-Lethal Doses of the Neonicotinoid Insecticide Imidacloprid.

Insecticides are a primary means for suppressing populations of insects that transmit plant pathogens. Application of insecticides for limiting the spread of insect-transmitted plant pathogens is often most effective when applied on an area-wide scale. The glassy-winged sharpshooter is a vector of the bacterial pathogen Xylella fastidiosa, which causes numerous plant diseases including Pierce's disease of grapevine. The glassy-winged sharpshooter has been the subject of an area-wide suppression program in California for nearly two decades. Overreliance on a limited number of active ingredients including the neonicotinoid imidacloprid has resulted in increased levels of resistance to commonly applied products. In California, glassy-winged sharpshooters move between citrus, an important overwintering host, and vineyards. Accordingly, imidacloprid is routinely applied via the irrigation system in vineyards and citrus orchards. For soil applied applications, it may take days to weeks for concentrations in plants to increase to lethal doses. Further, as the dose of imidacloprid required to kill sharpshooters increases due to resistance, so too does the period that sharpshooters are exposed to sub-lethal doses. Response of glassy-winged sharpshooter to cowpea plants treated with sub-lethal doses of imidacloprid was evaluated by conducting no-choice and choice tests. In no-choice feeding assays, glassy-winged sharpshooters caged on plants treated with sub-lethal doses of imidacloprid ceased feeding and produced little excreta. Further, sub-lethal exposure to a range of doses over a 4-d period did not affect viability over a 9-wk post-exposure holding period on untreated plants. In choice-tests, glassy-winged sharpshooters avoided treated plants and were observed predominately on untreated plants. Results suggest that application of imidacloprid to vineyards and citrus orchards may push glassy-winged sharpshooters out of treated habitats rather than kill them.

Evaluation of Alfalfa Fields and Pastures as Sources of Spissistilus festinus (Hemiptera: Membracidae): Quantification of Reproductive and Nutritional Parameters.

The threecornered alfalfa hopper (Spissistilus festinus) is a pest of grapevine, with damage caused by transmission of grapevine red blotch virus. Because grapevine is not a preferred host of the threecornered alfalfa hopper, abundance in vineyards depends on proximity to source habitats and presence of preferred hosts in vineyard understories. The potential for alfalfa fields and pastures in the Central Valley of California to serve as sources of threecornered alfalfa hopper was evaluated by quantifying parameters associated with threecornered alfalfa hopper reproductive and nutritional status. Laboratory studies determined that the threecornered alfalfa hopper is synovigenic, emerging as an adult prior to initiation of oogenesis and that females have multiple rounds of egg production. Alfalfa fields, irrigated pastures, and vineyards were sampled monthly. Adults were observed year-round in alfalfa fields and pastures, with populations peaking in fall. Gravid females were observed from February through November. While rare, adult threecornered alfalfa hoppers were collected from 2 of 4 sampled vineyards. In spring, adults were observed in samples collected from vineyard ground cover. In fall, adults were observed in samples collected from vineyard ground cover and foliage samples. Samples collected from pastures and vineyards were male biased, whereas equal numbers of males and females were observed in alfalfa fields. Adults collected from alfalfa fields were larger, heavier, and had greater estimated energetic reserves than adults collected from pastures. Adults collected from vineyards were of above average size and had relatively high estimated energetic reserves. Results suggest that alfalfa fields are more likely to serve as sources of threecornered alfalfa hoppers than irrigated pastures and that differences in male and female behavior may affect rates of pathogen transmission.

Mitigating an Epidemic of Resistance with Integrated Disease Management Tactics: Conflicting Management Recommendations from Insecticide Resistance and Epidemiological Models.

Insect-transmitted plant pathogens threaten crop production worldwide. Because a single feeding bout may be sufficient for a vector to transmit a pathogen that kills the plant, treatment thresholds for vectors of plant pathogens are low. For many vector species, overreliance on chemical controls has resulted in evolution of insecticide resistance. Analysis of complementary insecticide resistance and epidemiological models indicated that tactics for delaying resistance evolution conflict with tactics for limiting pathogen spread. Insecticide resistance models support maintaining untreated refuges that serve as a source of susceptible insects that reduce the likelihood of mating among rare resistant insects. In contrast, epidemiological models indicate that movement of vectors from untreated areas to insecticide-treated areas contributes to pathogen spread. Accordingly, epidemiological models support area-wide insecticide spray programs, although resistance models indicate that such an approach is likely to lead to rapid resistance. To mitigate risk of insecticide resistance, additional management approaches must be integrated into plant disease management strategies. The resistance and epidemiological models were used to evaluate effects of integrating application of insecticides with two additional management strategies: deployment of partially resistant plants (plants that are not immune to infection but have lower acquisition and inoculation rates than susceptible plants) and mating disruption (reduced vector birth rate in mating disruption-treated areas). Deployment of partially resistant plants reduced the risk that untreated areas served as a source of inoculative vectors. Mating disruption reduced the risk of resistance by suppressing growth of insecticide-resistant populations and benefited disease management by reducing vector abundance. Simulation results indicated that by targeting multiple aspects of the plant-pathogen-vector system, pathogen spread could be suppressed and resistance delayed. Implementation of such an approach will require innovations in vector control and sustained efforts in plant breeding.

Nutritional and Physiological Regulation of Glassy-Winged Sharpshooter Oogenesis.

The glassy-winged sharpshooter (Homalodisca vitripennis (Germar); Hemiptera: Cicadellidae: Cicadellinae) is an invasive insect that transmits the plant pathogenic bacterium Xylella fastidiosa Wells et al. (Xanthomonadales: Xanthomonadacae). While adult glassy-winged sharpshooter must feed to produce eggs, the role of nutritional status on initiating oogenesis is poorly understood. To determine the effects of glassy-winged sharpshooter nutrition on nymphal development, oogenesis, and fecundity, glassy-winged sharpshooter were reared on cowpea, sunflower, sorghum, and a mixture of the three plant species. Adults emerging from cowpea, sunflower, or plant mixture treatments had shorter development times, attained larger size, and had greater estimated lipid reserves than females reared on sorghum. In choice tests, nymphs avoided sorghum and preferentially fed on cowpea and sunflower. Adult females provisioned with a single plant species during the nymphal stage were provided with either the same host plant species or a mixture of host plant species (cowpea, sunflower, sorghum) for a 9-wk oviposition period, with 37% of females initiating oogenesis. Ovipositing females had greater juvenile hormone and octopamine levels than reproductively inactive females, although topical application of the juvenile hormone analog Methoprene did not promote oogenesis. Across nymphal diets, reproductively active females produced more eggs when held on plant mixtures than on single plant species. In choice tests, adult females were observed most frequently on cowpea, although most eggs were deposited on sorghum, the host least preferred by nymphs. Results suggest that fecundity is largely determined by the quality of the adult diet, although the stimulus that initiates oogenesis does not appear to be related to nutrition.

Spissistilus festinus (Hemiptera: Membracidae) susceptibility to six generalist predators.

Spissistilus festinus (Say) (Hemiptera: Membracidae) was shown to transmit Grapevine red blotch virus (GRBV) in a greenhouse study. Grapevines infected with GRBV exhibit reduced sugar accumulation, altered secondary metabolite production and delayed berry maturation that negatively impacts wine quality and economics. Augmentative biocontrol may be a useful integrated pest management (IPM) tool for suppressing S. festinus populations in vineyards, but minimal research has been conducted on testing potential predators against the different life stages of S. festinus. The susceptibility of S. festinus adults and nymphs (1st through 5th instar) to predation by six commercially available biocontrol agents in petri dish and bell bean plant arenas was determined under greenhouse conditions. No significant mortality of S. festinus nymphs or adults occurred when exposed to Cryptolaemus montrouzieri adults, C. montrouzieri larvae and Sympherobius barberi adults in petri dish or bell bean plant arenas. Significant mortality of 1st and 2nd instar nymphs of S. festinus in the presence of Zelus renardii nymphs was observed in petri dish but not in bell bean arenas. Hippodamia convergens adults and Chrysoperla rufilabris larvae both consumed a significant number of S. festinus nymphs in petri dish and bell bean arenas. No significant predation of S. festinus adults was documented in this experiment. Results of this study aid in identifying predators that may be suitable candidates for additional field testing to determine their potential efficacy as biocontrol agents of S. festinus in a vineyard setting.

Xylella fastidiosa and Glassy-Winged Sharpshooter Population Dynamics in the Southern San Joaquin Valley of California.

Xylella fastidiosa is a vector-transmitted bacterial plant pathogen that affects a wide array of perennial crops, including grapevines (Pierce's disease). In the southern San Joaquin Valley of California, epidemics of Pierce's disease of grapevine were associated with the glassy-winged sharpshooter, Homalodisca vitripennis. During the growing season, rates of X. fastidiosa spread in vineyards are affected by changes in pathogen distribution within chronically infected grapevines and by vector population dynamics. Grapevines chronically infected with X. fastidiosa rarely tested positive for the pathogen prior to July, suggesting vector acquisition of X. fastidiosa from grapevines increases as the season progresses. This hypothesis was supported by an increase in number of X. fastidiosa-positive glassy-winged sharpshooters collected from vineyards during July through September. Analysis of insecticide records indicated that vineyards in the study area were typically treated with a systemic neonicotinoid in spring of each year. As a result, abundance of glassy-winged sharpshooters was typically low in late spring and early summer, with abundance of glassy-winged sharpshooter adults increasing in late June and early July of each year. Collectively, the results suggest that late summer is a crucial time for X. fastidiosa secondary spread in vineyards in the southern San Joaquin Valley, because glassy-winged sharpshooter abundance, number of glassy-winged sharpshooters testing positive for X. fastidiosa, and grapevines with detectable pathogen populations were all greatest during this period.

Spread of Citrus Tristeza Virus in Citrus Orchards in Central California.

In California, citrus tristeza virus (CTV) is regulated by a State Interior Quarantine. In CTV abatement districts in central California, trees with CTV that react to MCA13 (MCA13-positive [MCA13+]), a strain-discriminating monoclonal antibody, are rogued to prevent virus spread. The Tulare County Pest Control District, however, does not participate in this abatement program except for a 1.6-km2 zone around the Lindcove Research and Extension Center, Exeter, CA. To quantify CTV spread under these two disparate management programs, CTV surveys were conducted in abatement plots with mandatory aphid control and nonabatement plots. Abatement plot surveys used hierarchical sampling of 25% of trees with samples pooled from four adjacent trees. Detection of MCA13+ CTV in a sample prompted resampling and testing of individual trees. From 2008 to 2018, incidence of CTV increased by an average of 3.9%, with only two MCA13+ samples detected. In contrast, in nonabatement plots, incidence of CTV increased by an average of 4.6% between 2015 and 2018. Increase in MCA13-negative (MCA-) isolates was 11 times greater than that of MCA13+ isolates, with the number of MCA13+ trees increasing by 19 trees between 2015 and 2018. MCA13- isolates were more randomly distributed, suggesting primary spread, whereas MCA13+ CTV isolates were more aggregated, suggesting some secondary spread. These results suggest that spread of MCA13+ isolates was limited by a combination of tree removal and aphid vector suppression. MCA13+ samples were VT isolates with some mixtures with T30 isolates. Despite the presence of VT isolates, all CTV-infected trees were asymptomatic.

Infection of Blueberry Cultivar 'Emerald' with a California Pierce's Disease Strain of Xylella fastidiosa and Acquisition by Glassy-Winged Sharpshooter.

Bacterial leaf scorch disease caused by Xylella fastidiosa occurs in southern highbush blueberry varieties in the southeastern United States. Susceptibility to X. fastidiosa varies by blueberry cultivar, and these interactions are often strain-specific. Xylella fastidiosa subsp. fastidiosa is the causal agent of Pierce's disease in grapevines, and it has been problematic in the San Joaquin Valley of California since the introduction of the glassy-winged sharpshooter (Homalodisca vitripennis). The glassy-winged sharpshooter is known to feed on blueberry, a crop that is expanding in the San Joaquin Valley. Currently, little is known about the potential for the spread of X. fastidiosa between grape and blueberry in this region. The ability of a Pierce's disease strain of X. fastidiosa from the San Joaquin Valley to cause disease in southern highbush blueberry and the potential for the glassy-winged sharpshooter to transmit X. fastidiosa between blueberry and grapevine were investigated. Experimental inoculations showed that the X. fastidiosa subsp. fastidiosa strain Bakersfield-1 can cause disease in blueberry cv. Emerald, and that the glassy-winged sharpshooter can acquire X. fastidiosa from artificially inoculated blueberry plants under laboratory conditions. Understanding the possibility for X. fastidiosa strains from the San Joaquin Valley to infect multiple crops grown in proximity is important for area-wide pest and disease management.

Effects of Nymphal Diet and Adult Feeding on Allocation of Resources to Glassy-Winged Sharpshooter Egg Production.

The glassy-winged sharpshooter is an invasive insect capable of transmitting the bacterial pathogen Xylella fastidiosa. Pre-oviposition periods of laboratory-reared glassy-winged sharpshooters are variable. Here, two questions were addressed: does nymphal diet affect pre-oviposition period and how do allocation patterns of resources differ for females that produce eggs versus females that do not? Nymphs were reared on one of three host plant species: cowpea, sunflower, or sorghum. Half of the females were sacrificed at emergence. The remaining adult females were held on cowpea, a host plant species known to support egg maturation via adult feeding. Females were sacrificed on the day of first oviposition or after 9 wk if no eggs were deposited. Females reared as nymphs on sorghum had longer development times and were smaller (head capsule width and hind tibia length) than females reared as nymphs on cowpea and sunflower. However, nymphal diet did not affect percentage of dry weight that was lipid at emergence. Further, nymphal diet did not affect time to deposition of the first egg mass or total number of eggs matured at the time of first oviposition. Egg production reduced the allocation of resources to insect bodies, with body lipid content decreasing with increasing egg production. In general, females increased wet weight 1.4-fold during the first week after adult emergence, with wet weights plateauing over the remaining 9 wk that adults were monitored. Thus, it seems reasonable to hypothesize that resources required for egg production were acquired via adult feeding during the first week after adult emergence.

Alfalfa and Pastures: Sources of Pests or Generalist Natural Enemies?

Pierce's disease of grapevine and almond leaf scorch disease are both caused by the bacterial pathogen Xylella fastidiosa Wells et al. In the Central Valley of California, Draeculacephala minerva Ball (Hemiptera: Cicadellidae) is the most common vector of X. fastidiosa. As alfalfa fields and pastures are considered source habitats for D. minerva, it is recommended that almond orchards and vineyards should be distanced from alfalfa and pastures. Here, risk of alfalfa and pastures serving as sources of D. minerva was compared to the potential benefit of alfalfa and pastures serving as sources of generalist natural enemies belonging to the families Chrysopidae and Coccinellidae. Populations of D. minerva were greatest in pastures, whereas chrysopids were least abundant in pastures, and coccinellids were only moderately more abundant in pastures than in vineyards or almond orchards. Accordingly, risk of pastures serving as a source of D. minerva was not offset by any potential benefit of pastures serving as a source of chrysopids or coccinellids. Abundance of D. minerva in alfalfa was low, whereas abundance of chrysopids and coccinellids in alfalfa was high. Thus, well-maintained alfalfa fields were a minor source of D. minerva that may contribute chrysopids and coccinellids to surrounding habitats. Spissistilus festinus (Say) (Hemiptera: Membracidae), a recently identified vector of grapevine red blotch virus, was abundant in alfalfa fields and was observed in vineyards. Thus, a full evaluation of the risk of planting vineyards near alfalfa may require considering risk associated with movement of S. festinus.

Effects of Energy Reserves and Diet on Glassy-Winged Sharpshooter Egg Maturation.

The glassy-winged sharpshooter is an invasive insect capable of transmitting the plant pathogen Xylella fastidiosa. As rates of pathogen spread are a function of vector abundance, identification of factors contributing to glassy-winged sharpshooter egg production will aid in predicting population growth. Here, effects of stored energy reserves and adult diet on glassy-winged sharpshooter egg maturation were evaluated. To estimate energy reserves available to adult females at the beginning of feeding assays, residuals from a regression of wet weight on size were used. Analysis of a subset of females sacrificed at the beginning of feeding assays, demonstrated that females with a positive residual wet weight had higher lipid content and carried more eggs than females with a negative residual wet weight. To evaluate effects of diet and energy reserves on egg maturation, energy reserves available to females entering feeding assays on cowpea and grapevine were estimated. For females held on cowpea, residual wet weight and quantity of excreta produced over a 6-d feeding period affected egg production. In contrast, for females held on grapevine, only residual wet weight affected egg production. Comparison of cowpea and grapevine xylem sap determined that eight amino acids were more concentrated in xylem sap from cowpea than from grapevine. Collectively, the results suggest that glassy-winged sharpshooter population growth within crop monocultures will not depend solely on the nutritional quality of the specific crop for producing mature eggs but also on the quantity of energy reserves accumulated by females prior to entering that crop habitat.

Effects of Xylem-Sap Composition on Glassy-Winged Sharpshooter (Hemiptera: Cicadellidae) Egg Maturation on High- and Low-Quality Host Plants.

Glassy-winged sharpshooters must feed as adults to produce mature eggs. Cowpea and sunflower are both readily accepted by the glassy-winged sharpshooter for feeding, but egg production on sunflower was reported to be lower than egg production on cowpea. To better understand the role of adult diet in egg production, effects of xylem-sap chemistry on glassy-winged sharpshooter egg maturation was compared for females confined to cowpea and sunflower. Females confined to cowpea consumed more xylem-sap than females held on sunflower. In response, females held on cowpea produced more eggs, had heavier bodies, and greater lipid content than females held on sunflower. Analysis of cowpea and sunflower xylem-sap found that 17 of 19 amino acids were more concentrated in cowpea xylem-sap than in sunflower xylem-sap. Thus, decreased consumption of sunflower xylem-sap was likely owing to perceived lower quality, with decreased egg production owing to a combination of decreased feeding and lower return per unit volume of xylem-sap consumed. Examination of pairwise correlation coefficients among amino acids indicated that concentrations of several amino acids within a plant species were correlated. Principal component analyses identified latent variables describing amino acid composition of xylem-sap. For females held on cowpea, egg maturation was affected by test date, volume of excreta produced, and principal components describing amino acid composition of xylem-sap. Principal component analyses aided in identifying amino acids that were positively or negatively associated with egg production, although determining causality with respect to key nutritional requirements for glassy-winged sharpshooter egg production will require additional testing.

Sequence polymorphism in an insect RNA virus field population: A snapshot from a single point in space and time reveals stochastic differences among and within individual hosts.

Population structure of Homalodisca coagulata Virus-1 (HoCV-1) among and within field-collected insects sampled from a single point in space and time was examined. Polymorphism in complete consensus sequences among single-insect isolates was dominated by synonymous substitutions. The mutant spectrum of the C2 helicase region within each single-insect isolate was unique and dominated by nonsynonymous singletons. Bootstrapping was used to correct the within-isolate nonsynonymous:synonymous arithmetic ratio (N:S) for RT-PCR error, yielding an N:S value ~one log-unit greater than that of consensus sequences. Probability of all possible single-base substitutions for the C2 region predicted N:S values within 95% confidence limits of the corrected within-isolate N:S when the only constraint imposed was viral polymerase error bias for transitions over transversions. These results indicate that bottlenecks coupled with strong negative/purifying selection drive consensus sequences toward neutral sequence space, and that most polymorphism within single-insect isolates is composed of newly-minted mutations sampled prior to selection.

Disentangling Effects of Vector Birth Rate, Mortality Rate, and Abundance on Spread of Plant Pathogens.

Models on the spread of insect-transmitted plant pathogens often fix vector population size by assuming that deaths are offset by births. Although such mathematical simplifications are often justified, deemphasizing parameters that govern vector population size is problematic, as reproductive biology and mortality schedules of vectors of plant pathogens receive little empirical attention. Here, the importance of explicitly including parameters for vector birth and death rates was evaluated by comparing results from models with fixed vector population size with models with logistic vector population growth. In fixed vector population size models, increasing vector mortality decreased percentage of inoculative vectors, but had no effect on vector population size, as deaths were offset by births. In models with logistic vector population growth, increasing vector mortality decreased percentage of inoculative vectors and decreased vector population size. Consequently, vector mortality had a greater effect on pathogen spread in models with logistic vector population growth than in models with fixed vector population size. Further, in models with logistic vector population growth, magnitude of vector birth rate determined time required for vector populations to reach large size, thereby determining when pathogen spread occurred quickly. Assumptions regarding timing of vector mortality within a time step also affected model outcome. A greater emphasis of vector entomologists on studying reproductive biology and mortality schedules of insect species that transmit plant pathogens will facilitate identification of conditions associated with rapid growth of vector populations and could lead to development of novel control strategies.

Effects of Glassy-Winged Sharpshooter (Hemiptera: Cicadellidae) Feeding, Size, and Lipid Content on Egg Maturation.

The glassy-winged sharpshooter (Homalodisca vitripennis) is synovigenic and must feed as an adult to produce eggs. Egg maturation rates depend on the host plant species provided to the adult female for feeding and are variable for females provided with the same host plant species. Here, the contribution of female size and lipid content to variation in egg maturation rates among females held on the same host plant species was assessed. To assess effects of female size and lipid content on egg maturation, feeding assays followed by measurements of egg load, female size, and lipid content were conducted. To accomplish this, females were field collected and held on cowpea until producing approximately 0, 12, 25, or 50 ml of excreta. After reaching prescribed excreta thresholds, females were dissected to determine egg load, hind tibia length, and head capsule width. Mature eggs were removed from the abdomen and dry weight of eggs and bodies (head, thorax, and abdomen) were obtained. Lipid content of eggs and bodies were determined using a quantitative colorimetric assay. Rates of body weight gain and body lipid gain were rapid with low levels of feeding (12 ml of excreta) but decelerated with additional feeding (>12 ml of excreta). In contrast, low levels of feeding (12 ml of excreta) resulted in little egg production, with rates of egg production accelerating with additional feeding (>12 ml of excreta). Accordingly, egg production was preceded by an increase in body dry weight and body lipid content. In agreement, probability that a female carried eggs increased with body lipid content in the 0-, 12-, and 25-ml feeding treatments. Across treatments, larger females carried more eggs than smaller females. Collectively, results suggest that variation in glassy-winged sharpshooter egg maturation rates partially may be explained by availability of lipid reserves at the start of a feeding bout and female size.

Evaluation of a method to quantify glassy-winged sharpshooter (Hemiptera: Cicadellidae) egg maturation during a feeding assay.

A method to improve an assay relating adult feeding to egg maturation by the glassy-winged 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.

Evaluation of Olive as a Host of Xylella fastidiosa and Associated Sharpshooter Vectors.

Olive (Olea europaea) trees exhibiting leaf scorch or branch dieback symptoms in California were surveyed for the xylem-limited, fastidious bacterium Xylella fastidiosa. Only approximately 17% of diseased trees tested positive for X. fastidiosa by polymerase chain reaction, and disease symptoms could not be attributed to X. fastidiosa infection of olive in greenhouse pathogenicity assays. Six strains of X. fastidiosa were isolated from olive in Southern California. Molecular assays identified strains recovered from olive as belonging to X. fastidiosa subsp. multiplex. Pathogenicity testing of olive strains on grapevine and almond confirmed that X. fastidiosa strains isolated from olive yield disease phenotypes on almond and grapevine typical of those expected for subsp. multiplex. Mechanical inoculation of X. fastidiosa olive strains to olive resulted in infection at low efficiency but infections remained asymptomatic and tended to be self-limiting. Vector transmission assays demonstrated that glassy-winged sharpshooter (Homalodisca vitripennis) could transmit strains of both subspp. multiplex and fastidiosa to olive at low efficiency. Insect trapping data indicated that two vectors of X. fastidiosa, glassy-winged sharpshooter and green sharpshooter (Draeculacephala minerva), were active in olive orchards. Collectively, the data indicate that X. fastidiosa did not cause olive leaf scorch or branch dieback but olive may contribute to the epidemiology of X. fastidiosa-elicited diseases in California. Olive may serve as an alternative, albeit suboptimal, host of X. fastidiosa. Olive also may be a refuge where sharpshooter vectors evade intensive areawide insecticide treatment of citrus, the primary control method used in California to limit glassy-winged sharpshooter populations and, indirectly, epidemics of Pierce's disease of grapevine.

Distinguishing Nonpareil marketing group almond cultivars through multivariate analyses.

More than 80% of the world's almonds are grown in California with several dozen almond cultivars available commercially. To facilitate promotion and sale, almond cultivars are categorized into marketing groups based on kernel shape and appearance. Several marketing groups are recognized, with the Nonpareil Marketing Group (NMG) demanding the highest prices. Placement of cultivars into the NMG is historical and no objective standards exist for deciding whether newly developed cultivars belong in the NMG. Principal component analyses (PCA) were used to identify nut and kernel characteristics best separating the 4 NMG cultivars (Nonpareil, Jeffries, Kapareil, and Milow) from a representative of the California Marketing Group (cultivar Carmel) and the Mission Marketing Group (cultivar Padre). In addition, discriminant analyses were used to determine cultivar misclassification rates between and within the marketing groups. All 19 evaluated carpological characters differed significantly among the 6 cultivars and during 2 harvest seasons. A clear distinction of NMG cultivars from representatives of the California and Mission Marketing Groups was evident from a PCA involving the 6 cultivars. Further, NMG kernels were successfully discriminated from kernels representing the California and Mission Marketing Groups with overall kernel misclassification of only 2% using 16 of the 19 evaluated characters. Pellicle luminosity was the most discriminating character, regardless of the character set used in analyses. Results provide an objective classification of NMG almond kernels, clearly distinguishing them from kernels of cultivars representing the California and Mission Marketing Groups.