The Effects of Constant and Fluctuating Temperatures on Development of Diaphorina citri (Hemiptera: Liviidae), the Asian Citrus Psyllid.

The effects of six average daily temperatures, 15, 20, 25, 30, 32, and 35°C, that were either constant or fluctuating over 24 h on development times of California-sourced Diaphorina citri Kuwayama nymphs were examined. Thermal performance curves for immature stages of D. citri were characterized using one linear and six nonlinear models (i.e., Ratkowsky, Lobry-Rosso-Flandrois, Lactin-2, Brière-2, Beta, and Performance-2). Daily thermal fluctuations had significant effects on development times of D. citri nymphs, which differed across experimental temperatures. Diaphorina citri nymphs reared at constant temperatures completed development faster than those reared under fluctuating profiles with equivalent temperature means. Linear model estimates of degree-days required for completion of cumulative development of D. citri were 25% lower for constant temperatures when compared with fluctuating temperature regimens. Nonlinear model estimations of optimum developmental temperature and upper theoretical temperature bounds for development were similar for individuals reared under constant and fluctuating temperatures. Nevertheless, the estimated values of lower theoretical temperature limits above which development occurred were lower under fluctuating than constant temperatures. A meta-analysis of published D. citri temperature-dependent development literature, synthesizing datasets of five globally distributed populations (Brazil, California, China, Florida, and Japan) reared under different constant temperatures on six different host plants (i.e., Citrus limonia, C. sinensis cv Natal, C. sinensis cv. Pêra, C. reticulata, Fortunella margarita, and Murraya paniculata), together with the results of this study (C. volkameriana), revealed convergence in estimates of developmental parameters. These results have implications for predicting D. citri invasion and establishment risk and subsequent population performance across various climactic gradients and geographic regions.

Spatiotemporal colonization of Xylella fastidiosa in its vector supports the role of egestion in the inoculation mechanism of foregut-borne plant pathogens.

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 laser-scanning 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.

Estimation of feeding threshold for Homalodisca vitripennis (Hemiptera: Cicadellidae) and its application to prediction of overwintering mortality.

The glassy-winged sharpshooter, Homalodisca vitripennis (Germar), vectors the bacterium Xylella fastidiosa that induces Pierce's disease of grape. This study determined the effect of temperature on the feeding activity of H. vitripennis adults and the resulting production of excreta. The Logan type I model described a nonlinear pattern that showed excreta production increased up to an optimal temperature (33.1°C), followed by an abrupt decline near an estimated upper threshold (36.4°C). A temperature threshold for feeding, at or below which adults cease feeding, was estimated to be 10°C using a linear regression model based on the percentage of adults producing excreta over a range of constant temperatures. A simulated winter-temperature experiment using fluctuating thermal cycles confirmed that a time period above the temperature threshold for feeding was a critical factor in determining adult survival. Using data from the simulated temperature study, a predictive model was constructed by quantifying the relationship between cumulative mortality and cooling degree-hours. In field validation experiments, the model accurately predicted the temporal pattern of overwintering mortality of H. vitripennis adults held under winter temperatures simulating conditions in Bakersfield and Riverside, California, in 2006-2007. Model prediction using winter temperature data from a Riverside weather station indicated that H. vitripennis adults would experience an average of 92% overwintering mortality before reproduction in the spring, but levels of mortality varied depending on winter temperatures. The potential for temperature-based indices to predict temporal and spatial dynamics of H. vitripennis overwintering is discussed.

Influences of temperature on Homalodisca vitripennis (Hemiptera: Cicadellidae) survival under various feeding conditions.

Survival of the glassy-winged sharpshooter, Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae), was studied under various constant temperatures and feeding conditions. When provided a host plant (Citrus limon L. Burm. f.) to feed on during a 21-d trial, 100% mortality occurred at 0.1, 3.2, and 40.1 degrees C, whereas an average of 74-76% of adults survived in the 13.2-24.5 degrees C range. When individually confined with moist cotton, adult longevity was greatest (16.3 d) at 13.3 degrees C, but it was <3 d at -2.4 and 36.2 degrees C. In a companion study comparing the presence versus absence of a host plant, the presence of a host plant was not a significant factor influencing survival at temperatures < or =7.8 degrees C but was at temperatures > or =18.9 degrees C. The relationship between temperature and survival was described by a nonlinear function that estimated the optimum temperature in each feeding regimen: no host plant or moist cotton (5.5 degrees C), moist cotton (9.9 degrees C), and accessible host plant (25.1 degrees C). The model quantitatively predicted that H. vitripennis would survive longer periods at a wider temperature regimen when provided with a host plant than when provided with water alone (moist cotton) or when provided with neither plant host nor water. Our results suggest that continuous exposure to either low (<5 degrees C) or high (>30 degrees C) temperatures are detrimental for adult survival. Specifically, low temperatures caused early mortality because of inhibition of feeding activity and presumably this threshold lies between 7.8 and 13.2 degrees C. Furthermore, this study clearly shows that temperature may influence the survival of H. vitripennis adults regardless of feeding regimens, and its implications for population dynamics are discussed.

Molecular distinction between populations of Gonatocerus morrilli, egg parasitoids of the glassy-winged sharpshooter from Texas and California: do cryptic species exist?

Two molecular methods were utilized to distinguish geographic populations of Gonatocerus morrilli (Howard) from Texas and California and to test the possibility that this species could exist as a species-complex. Inter-Simple Sequence Repeat-Polymerase Chain Reactions (ISSR-PCR) were performed with a 5'-anchored ISSR primer. Twenty-five markers were generated with four populations (40 individuals) of G. morrilli. Twenty-three were polymorphic and the percentage of polymorphic loci was 92%. Most markers could be considered diagnostic since there was no band sharing between the Texas and California populations. Such differences typically are not found unless the populations are reproductively isolated. Exact tests for population differentiation indicated significant differences in marker frequencies among the populations. Comparison of other genetic differentiation estimates, which evaluate the degree of genetic subdivision, demonstrated excellent agreement between GST and theta values, 0.92 and 0.94, respectively, indicating that about 92 to 94% of the variance was distributed among populations. The average genetic divergence (D), as measured by genetic distance, was extremely high (Nei = 0.82 and Reynolds = 2.79). A dendrogram based on Nei's genetic distance separated the Texas and California populations into two clusters, respectively. Amplification of the Internal Transcribed Spacer-1 (ITS-1) region showed no size differences, whereas the ITS-2 DNA fragment varied in size between the two geographic populations. The ITS-2 fragment sizes were about 865 and 1099 base pairs for the California and Texas populations, respectively. The present study using the two molecular methods provides novel data critical to the glassy-winged sharpshooter/Pierce's disease biological control program in California.