Effect of selected insecticides on Homalodisca coagulata (Homoptera: Cicadellidae) and transmission of oleander leaf scorch in a greenhouse study.

Homalodisca coagulata (Say) is a recent introduction to California. It is known to spread a strain of the bacterium Xylella fastidiosa Wells, Raju, Hung, Weisberg, Mandelco-Paul & Brenner that induces oleander leaf scorch disease in oleander, Nerium oleander L. Oleander leaf scorch is lethal to oleander and threatens to decimate one of the most important landscape shrubs in California. Towards developing a management strategy for H. coagulata-spread oleander leaf scorch, we documented the affects of selected insecticides on H. coagulata mortality, feeding behavior, and disease transmission in a greenhouse study. Oleanders treated with fenpropathrin, fenpropathrin + acephate, and imidacloprid caused significant mortality to caged H. coagulata within 4 h of exposure. Within 24 h, these pesticides caused nearly 100% mortality 3 wk after treatment. In other experiments, acetamiprid and fenpropathrin treatments reduced time spent feeding and total time on plants. H. coagulata on fenpropathrin-, acetamiprid-, and imidacloprid-treated oleander died in less than 13 min on average. Oleander leaf scorch transmission by H. coagulata was blocked by applications of foliar-applied acetamiprid, and soil-applied imidacloprid and thiamethoxam.     

Biochemical monitoring of acetylcholinesterase sensitivity to organophosphorus insecticides in glassy-winged sharpshooter Homalodisca coagulata Say (Homoptera: Cicadellidae) and smoke-tree sharpshooter H. lacerta Fowler

The glassy-winged sharpshooter Homalodisca coagulata Say (Homoptera: Cicadellidae) is a new pest to California agriculture. It is the principal vector of several plant pathogenic diseases, particularly Pierce's Disease in grapevines, and oleander leaf scorch. A microplate-based assay is described that measures the sensitivity of acetylcholinesterase (AChE) activity to inhibition by organophosphorus (OP) insecticides in this important pest. The technique provides users with an accurate measure of the efficacy of OP binding to this target site, and is a valuable tool for monitoring field populations of the insect to determine whether the use of OP insecticides has selected for resistant individuals. The technique will also measure AChE sensitivity in the smoke-tree sharpshooter, H. lacerta Fowler. This species is native to California and is regarded only as a minor pest. Both inhibition and kinetic measurements for the AChE enzymes in these sharpshooters demonstrate the close phylogenetic relationships between the two species.     

Detection and differentiation of Xylella fastidiosa strains acquired and retained by glassy-winged sharpshooters (Hemiptera: Cicadellidae) using a mixture of strain-specific primer sets

Xylella fastidiosa Wells is a bacterial pathogen that causes a variety of plant diseases, including Pierce's disease (PD) of grapevine, almond leaf scorch, alfalfa dwarf, citrus variegated chlorosis, and oleander leaf scorch (OLS). Numerous strains of this pathogen have been genetically characterized, and several different strains occur in the United States. The dominant vector in southern California is the glassy-winged sharpshooter, Homalodisca coagulata (Say) (Hemiptera: Cicadellidae). The high mobility of this insect, and its use of large numbers of host plant species, provides this vector with ample exposure to multiple strains of X. fastidiosa during its lifetime. To learn more about the ability of this vector to acquire, retain, and transmit multiple strains of the pathogen, we developed a polymerase chain reaction (PCR)-based method to detect and differentiate strains of X. fastidiosa present in individual glassy-winged sharpshooter adults. Insects were sequentially exposed to plants infected with a PD strain in grapevine and an OLS strain in oleander. After sequential exposure, a few insects tested positive for both strains (7%); however, in most cases individuals tested positive for only one strain (29% PD, 41% OLS). In transmission studies, individual adults transmitted either the PD or OLS strain of the pathogen at a rate (39%) similar to that previously reported after exposure to a single strain, but no single individual transmitted both strains of the pathogen. PD and OLS strains of X. fastidiosa remained detectable in glassy-winged sharpshooter, even when insects were fed on a plant species that was not a host of the strain for 1 wk.     

Transmission of Xylella fastidiosa to grapevines by Homalodisca coagulata (Hemiptera: Cicadellidae)

Pierce's disease (PD) of grapevines is caused by a xylem-limited bacterium Xylella fastidiosa (Wells, Raju, Hung, Weisburg, Mandelco-Paul, and Brenner) that is transmitted to plants by xylem sap-feeding insects. The introduction of the sharpshooter leafhopper Homalodisca coagulata (Say) into California has initiated new PD epidemics in southern California. In laboratory experiments, the major characteristics of H. coagulata's transmission of X. fastidiosa to grapevines were the same as reported for other vectors: short or absent latent period; nymphs transmitted but lost infectivity after molting and regained infectivity after feeding on infected plants; and infectivity persisted in adults. Adult H. coagulata acquired and inoculated X. fastidiosa in <1 h of access time on a plant. Inoculation rates increased with access time, but acquisition efficiency (20% per individual) did not increase significantly beyond 6-h access. Estimated inoculation efficiency per individual per day was 19.6, 17.9, and 10.3% for experiments where plant access was 1, 2, and 4 d, respectively. Freshly molted adults and nymphs acquired and transmitted X. fastidiosa more efficiently than did older, field-collected insects. H. coagulata transmitted X. fastidiosa to 2-yr-old woody tissues of grapevines as efficiently as to green shoots. H. coagulata transmitted X. fastidiosa 3.5 mo after acquisition, demonstrating persistence of infectivity in adults. About half (14/29) of the H. coagulata from which we failed to culture X. fostidiosa from homogenized heads (with a detection threshold of 265 CFU/head) transmitted the pathogen to grape, and 17 of 24 from which we cultured X. fastidiosa transmitted.     

Dispersion of Homalodisca coagulata (Hemiptera: Cicadellidae), a vector of Xylella fastidiosa, into vineyards in southern California

Recent epidemics of Pierce's disease of grapevine in California vectored by Homalodisca coagulata (Say), an invasive vector species, have characteristics that differ from epidemics involving native vectors. Among these differences are the longer distances and greater speed that the disease is spread by H. coagulata. In this investigation, we used yellow sticky traps to study the seasonal dispersion activity of H. coagulata in a southern California grape-growing area in which an epidemic of Pierce's disease has caused large losses. For 21 mo, we monitored adult H. coagulata at the edges of vineyards bordering citrus, an important crop host, natural coastal sage scrub vegetation, and natural riparian vegetation. We also monitored H. coagulata dispersion from 0 to 40 m into vineyards. Finally, we examined the vertical dispersion of H. coagulata adults into grapevines through a season. This investigation showed that H. coagulata is associated with citrus, from where it disperses deep into vineyards, and not just the vineyard edge as with Pierce's disease vectors that are native to California. Peak dispersion into vineyards occurred in the summer. Another period of H. coagulata activity occurred in the winter in vineyards bordering citrus. Through the period of peak flight activity, 97% of all H. coagulata adults trapped between 1 and 7 m were caught at an altitude of 5 m or lower, suggesting the potential of a barrier as a management tactic to keep H. coagulata out of vineyards.     

Feasibility of tracking within-field movements of Homalodisca coagulata (Hemiptera: Cicadellidae) and estimating its densities using fluorescent dusts in mark-release-recapture experiments

A mark-release-recapture technique was developed and tested for use in tracking the field movements of adult glassy-winged sharpshooters, Homalodisca coagulata (Say) (Hemiptera: Cicadellidae), in various agricultural and urban plantings. Greenhouse experiments in which adult H. coagulata were marked with one of five colored fluorescent dusts (Aurora Pink-All, Horizon Blue-A19, Blaze Orange-A15N, Saturn Yellow-A17, and Corona Magenta-A21) and released into cages with citrus seedlings showed that their mortality rates during a 30-d period were statistically similar to that of the undusted controls. Adults marked with a sixth dust color (Signal Green-A18N) suffered higher rates of mortality than did the undusted controls and thus were eliminated from further consideration. Adult H. coagulata marked with one of the five accepted colors of fluorescent dust were able to fly beyond 100 m in a field devoid of vegetation within minutes of their release, and the marking did not affect overall flight behavior significantly compared with that of the undusted controls. However, at wind speeds above 5 m/s, percentage recapture was significantly reduced, which indicates that both dusted and undusted adults were unable to orient their flight. In total, 41,124 marked and unmarked adults were released in the three field experiments in southern California (Riverside and Kern counties) during 2000 and 2001 to evaluate flight dispersal and estimate densities of H. coagulata. The mark-release-recapture and feral data obtained during the June, July, and August 2001 studies, when coupled with the Lincoln index, yielded estimates of adult H. coagulata of 1.2 and 2.2 million per ha, respectively, at a San Joaquin Valley (Kern Co.) and a southern California (Riverside Co.) citrus grove. The use of colored dusts to mark H. coagulata proved to be reliable, cost-effective, and time-efficient for mark-release-recapture studies with this insect within a citrus grove, but they are less likely to be useful for studies of adult H. coagulata movements among plantings.     

Spatial and temporal dynamics of overwintering Homalodisca coagulata (Hemiptera: Cicadellidae)

A 4-yr landscape-scale study was conducted to investigate spatial and temporal dynamics of overwintering Homalodisca coagulata (Say) (Hemiptera: Cicadellidae) in the lower San Joaquin Valley, California. Spatial structures of H. coagulata distributions were characterized with Moran's I index, and spatial associations between H. coagulata and the surrounding environment were investigated with a geographic information system. H. coagulata was caught consistently with sticky traps throughout the winter, and trap catches formed a distinctive peak in December or January, indicating active flight of H. coagulata during the winter. In 2000-2001, the mean +/-SE trap count was 4.8 +/- 1.21 per trap per wk, and H. coagulata trap catches were spatially autocorrelated within approximately 1.3 km. Approximately 49% of H. coagulata were caught in citrus, 23% in stone fruit, and 11% in grape. After a control program began in spring 2001, the mean trap count was considerably lower (0.041 +/- 0.0004 per trap per wk), and no spatial autocorrelations were detected in 2001-2004. H. coagulata trap catch-crop associations also changed after initiation of the control program. Between 25 and 38% of H. coagulata trap catches were from citrus, between 8 and 20% were from stone fruit, and between 11 and 25% were from grape. Potential for winter-season spread and management of Xylella fastidiosa Wells et al., a pathogen causing Pierce's disease, are discussed.     

Correlation of stylet activities by the glassy-winged sharpshooter, Homalodisca coagulata (Say), with electrical penetration graph (EPG) waveforms

Glassy-winged sharpshooter, Homalodisca coagulata (Say), is an efficient vector of Xylella fastidiosa (Xf), the causal bacterium of Pierce's disease, and leaf scorch in almond and oleander. Acquisition and inoculation of Xf occur sometime during the process of stylet penetration into the plant. That process is most rigorously studied via electrical penetration graph (EPG) monitoring of insect feeding. This study provides part of the crucial biological meanings that define the waveforms of each new insect species recorded by EPG. By synchronizing AC EPG waveforms with high-magnification video of H. coagulata stylet penetration in artifical diet, we correlated stylet activities with three previously described EPG pathway waveforms, A1, B1 and B2, as well as one ingestion waveform, C. Waveform A1 occured at the beginning of stylet penetration. This waveform was correlated with salivary sheath trunk formation, repetitive stylet movements involving retraction of both maxillary stylets and one mandibular stylet, extension of the stylet fascicle, and the fluttering-like movements of the maxillary stylet tips. Waveform B1 was ubitquious, interspersed throughout the other waveforms. B1 sub-type B1w was correlated with salivation followed by maxillary tip fluttering. This tip fluttering also occurred before and during B1 sub-type B1s, but was not directly correlated with either the occurrence or frequency of this waveform. Waveform B2 was correlated with sawing-like maxillary stylet movements, which usually occurred during salivary sheath branching. Waveform C was correlated with ingestion. Fluid outflow was also observed as a mechanism to clear the maxillary tips from debris during waveform C. This detailed understanding of stylet penetration behaviors of H. coagulata is an important step toward identifying the instant of bacterial inoculation which, in turn, will be applied to studies of disease epidemiology and development of host plant resistance.     

Establishment of baseline susceptibility data to various insecticides for Homalodisca coagulata (Homoptera: Cicadellidae) by comparative bioassay techniques

Homalodisca coagulata Say, adults from three locations in California were subjected to insecticide bioassays to establish baseline toxicity. Initially, two bioassay techniques, petri dish and leaf dip, were compared to determine the most useful method to establish baseline susceptibility data under laboratory and greenhouse conditions. Comparative dose-response data were determined by both techniques to endosulfan, dimethoate, cyfluthrin, and acetamiprid. Toxic values were similar to some insecticides with both techniques but not for all insecticides, revealing susceptibility differences among the three populations of H. coagulata. In subsequent tests, the petri dish technique was selected to establish baseline susceptibility data to various contact insecticides. A systemic uptake bioassay was adapted to estimate dose-mortality responses to a systemic insecticide, imidacloprid. A 2-yr comparison of toxicological responses showed all three populations of H. coagulata to be highly susceptible to 10 insecticides, including chlorpyrifos, dimethoate, endosulfan, bifenthrin, cyfluthrin, esfenvalerate, fenpropathrin, acetamiprid, imidacloprid, and thiamethoxam. In general, two pyrethroids, bifenthrin and esfenvalerate, were the most toxic compounds, followed by two neonicotinoids, acetamiprid and imidacloprid. The LC50 values for all insecticides tested were lower than concentrations used as recommended field rates. Baseline data varied for the three geographically distinct H. coagulata populations with the petri dish technique. Adult H. coagulata collected from San Bernardino County were significantly more susceptible to select pyrethroids compared with adults from Riverside or Kern counties. Adults from San Bernardino County also were more sensitive to two neonicotinoids, acetamiprid and imidacloprid. The highest LC50 values were to endosulfan, which nonetheless proved highly toxic to H. coagulata from all three regions. In the majority of the tests, mortality increased over time resulting in increased susceptibility at 48 h compared with 24 h. These results indicate a wide selection of highly effective insecticides that could aid in managing H. coagulata populations in California.     

Transmission efficiency of Xylella fastidiosa by sharpshooters (Hemiptera: Cicadellidae) in coffee and citrus

Xylella fastidiosa (Wells, Raju, Hung, Weisburg, Mandelco-Paul, and Brenner) is a bacterial pathogen transmitted by several sharpshooters in two tribes of Cicadellinae (Proconiini and Cicadellini). Here, we compared the transmission efficiency of X. fastidiosa in coffee (Coffea arabica L.) and citrus [Citrus sinensis (L.) Osbeck] by Cicadellini [Bucephalogonia xanthophis (Berg) and Dilobopterus costalimai Young] and Proconiini [Homalodisca ignorata Melichar and Oncometopia facialis (Signoret)] sharpshooters that occur in both crops. At different seasons, healthy adults of each species were submitted to a 48-h acquisition access period on citrus or coffee source plants infected with X. fastidiosa isolates that cause Citrus variegated chlorosis (CVC) and Coffee leaf scorch (CLS), respectively, and then confined on healthy seedlings of the corresponding host plant for a 48-h inoculation access period. No significant effect of inoculation season was observed when comparing infection rates of citrus or coffee plants inoculated by vectors at different times of the year. In citrus, the transmission rate by single insects was significantly higher for H. ignorata (30%) in relation to B. xanthophis (5%) and O. facialis (1.1%), but there was no difference among vector species in coffee, whose transmission rates ranged from 1.2 to 7.2%. Comparing host plants, H. ignorata was more effective in transmitting X. fastidiosa to citrus (30%) in relation to coffee (2.2%), whereas the other vectors transmitted the bacterium to both hosts with similar efficiencies. Despite these variations, vector efficiency in coffee and citrus is lower than that reported in other hosts.     

Evaluation of methods for extracting Xylella fastidiosa DNA from the glassy-winged sharpshooter

The recent spread of the plant pathogenic bacterium Xylclla fastidiosa Wells et al. by an invasive vector species, Homalodisca coagulata Say, in southern California has resulted in new epidemics of Pierce's disease of grapevine. Our goal is to develop an efficient method to detect low titers of X. fastidiosa in H. coagulata that is amenable to large sample sizes for epidemiological studies. Detection of the plant pathogenic bacterium X. fastidiosa in its insect vector is complicated by low titers of bacteria, difficulty in releasing it from the insect mouthparts and foregut, and the presence of substances in the insect that inhibit polymerase chain reaction (PCr). To select the optimal protocol for DNA extraction to be used with PCR, we compared three standard methods and 11 commercially available kits for relative efficiency of X. fastidiosa DNA extraction in the presence of insect tissue. All of the protocols tested were proficient at extracting DNA from pure bacterial culture (1 x 10(5) cells), and all but one protocol successfully extracted sufficient bacterial DNA in the presence of insect tissue. Three DNA extraction techniques, immunomagnetic separation, the DNeasy Tissue kit (Qiagen, Hercules, CA), and Genomic DNA Purification kit (Fermentus, Hanover, MD), were compared more closely using a dilution series of X. fastidiosa (5000-0 cells) with and without insect tissue present. The DNeasy Tissue kit was the best kit tested, allowing detection of 5 x 10(3) X. fastidiosa cells with an insect head background.     

Genetic diversity of Pierce's disease strains and other pathotypes of Xylella fastidiosa

Strains of Xylella fastidiosa isolated from grape, almond, maple, and oleander were characterized by enterobacterial repetitive intergenic consensus sequence-, repetitive extragenic palindromic element (REP)-, and random amplified polymorphic DNA (RAPD)-PCR; contour-clamped homogeneous electric field (CHEF) gel electrophoresis; plasmid content; and sequencing of the 16S-23S rRNA spacer region. Combining methods gave greater resolution of strain groupings than any single method. Strains isolated from grape with Pierce's disease (PD) from California, Florida, and Georgia showed greater than previously reported genetic variability, including plasmid contents, but formed a cluster based on analysis of RAPD-PCR products, NotI and SpeI genomic DNA fingerprints, and 16S-23S rRNA spacer region sequence. Two groupings of almond leaf scorch (ALS) strains were distinguished by RAPD-PCR and CHEF gel electrophoresis, but some ALS isolates were clustered within the PD group. RAPD-PCR, CHEF gel electrophoresis, and 16S-23S rRNA sequence analysis produced the same groupings of strains, with RAPD-PCR resolving the greatest genetic differences. Oleander strains, phony peach disease (PP), and oak leaf scorch (OLS) strains were distinct from other strains. DNA profiles constructed by REP-PCR analysis were the same or very similar among all grape strains and most almond strains but different among some almond strains and all other strains tested. Eight of 12 ALS strains and 4 of 14 PD strains of X. fastidiosa isolated in California contained plasmids. All oleander strains carried the same-sized plasmid; all OLS strains carried the same-sized plasmid. A plum leaf scald strain contained three plasmids, two of which were the same sizes as those found in PP strains. These findings support a division of X. fastidiosa at the subspecies or pathovar level.     

16S rDNA sequence analysis of Xylella fastidiosa strains

The 16S rDNA encoding the small subunit ribosomal RNA were amplified by PCR, cloned, and sequenced from 16 strains of Xylella fastidiosa originating from nine different hosts. In pair-wise comparisons, X. fastidiosa strains showed a maximum variation of 1.0% or 14 nucleotide positions. When all 16 sequences were considered as a set, 54 variable positions were found. Analysis of the sequence data indicated that the X. fastdiosa strains formed three rDNA groups. Group one includes Pierce's disease and mulberry leaf scorch strains; Group two, periwinkle wilt, plum leaf scald, phony peach, oak leaf scorch, and elm leaf scorch strains; and Group three, citrus variegated chlorosis and coffee leaf scorch strains. All X. fastidiosa strains exhibited significantly higher levels of sequence heterogeneity (63 to 83 nucleotide positions) when compared to species from Xanthomonas and Stenotrophomonas. Our data demonstrate that 16S rDNA sequence data could provide valuable information for future classification of X. fastidiosa at the sub-species level.     

Fate of a genetically modified bacterium in foregut of glassy-winged sharpshooter (Hemiptera: Cicadellidae)

Symbiotic control is a new strategy being investigated to prevent the spread of insect-transmitted pathogens by reducing vector competence. We are developing this strategy to reduce the spread of Xylella fastidiosa by Homalodisca vitripennis (Germar) [formerly Homalodisca coagulata (Say)] (Hemiptera: Cicadellidae), the glassy-winged sharpshooter. In this study, the fate of a transformed symbiotic bacterium, Alcaligenes xylosoxidans variety denitriicans (S1Axd), in the foregut of glassy-winged sharpshooter when fed on citrus (Citrus spp.) and grape (Vitris spp.) was assessed. TaqMan-based quantitative real-time polymerase chain reaction (PCR) was used to detect and quantify bacterial cells remaining in the foregut at 0, 2, 4, 9, and 12 d after acquisition. S1Axd titer dropped rapidly by 2 d after acquisition, but in spite of this, at end of the 12-d experimental period, 45 and 38% of the glassy-winged sharpshooters retained the transformed bacteria, when fed on grape and citrus, respectively.     

A SYBR green-based real-time polymerase chain reaction protocol and novel DNA extraction technique to detect Xylella fastidiosa in Homalodisca coagulata

Homalodisca coagulata Say (Hemiptera: Cicadellidae) is a major agronomic pest because it transmits Xylella fastidiosa (Wells), the bacterium that causes Pierce's disease of grapevine. The ability to easily detect X. fastidiosa in populations of H. coagulata facilitates epidemiological studies and development of a monitoring program supporting disease management. Such a program depends on a detection protocol that is rapid, reproducible, and amenable to large sample sizes, while remaining sensitive enough to detect low amounts of pathogen DNA. In this study, we developed an improved method to speed DNA extraction by implementing a simple vacuum step that replaces labor- and time-intensive maceration of tissue samples and that is compatible with manufactured DNA extraction kits. Additionally, we have developed a SYBR Green-based real-time (RT)-polymerase chain reaction (PCR) system, which uses traditional PCR primers that are relatively inexpensive and effective. Using this extraction/RT-PCR system, we found no statistically significant differences in the detection of X. fastidiosa among samples that were either immediately extracted or stored dry or in mineral oil for 10 d at -4 degrees C. In further testing, we found no significant reduction in detection capabilities for X. fastidiosa-fed H. coagulata left in the sun on yellow sticky cards for up to 6 d. Therefore, we recommend a field-based detection system that includes recovery of H. coagulata from sticky traps for up to 6 d after trapping, subsequent freezing of samples for as long as 10 d before vacuum extraction is performed, and detection of the bacterium by SYBR Green-based RT-PCR.     

Multilocus Simple Sequence Repeat Markers for Differentiating Strains and Evaluating Genetic Diversity of Xylella fastidiosa

A genome-wide search was performed to identify simple sequence repeat (SSR) loci among the available sequence databases from four strains of Xylella fastidiosa (strains causing Pierce's disease, citrus variegated chlorosis, almond leaf scorch, and oleander leaf scorch). Thirty-four SSR loci were selected for SSR primer design and were validated in PCR experiments. These multilocus SSR primers, distributed across the X. fastidiosa genome, clearly differentiated and clustered X. fastidiosa strains collected from grape, almond, citrus, and oleander. They are well suited for differentiating strains and studying X. fastidiosa epidemiology and population genetics.     

A multigene phylogenetic study of clonal diversity and divergence in North American strains of the plant pathogen Xylella fastidiosa

Xylella fastidiosa is a pathogen that causes leaf scorch and related diseases in over 100 plant species, including Pierce's disease in grapevines (PD), phony peach disease (PP), plum leaf scald (PLS), and leaf scorch in almond (ALS), oak (OAK), and oleander (OLS). We used a high-resolution DNA sequence approach to investigate the evolutionary relationships, geographic variation, and divergence times among the X. fastidiosa isolates causing these diseases in North America. Using a large data set of 10 coding loci and 26 isolates, the phylogeny of X. fastidiosa defined three major clades. Two of these clades correspond to the recently identified X. fastidiosa subspecies piercei (PD and some ALS isolates) and X. fastidiosa subsp. multiplex (OAK, PP, PLS, and some ALS isolates). The third clade grouped all of the OLS isolates into a genetically distinct group, named X. fastidiosa subsp. sandyi. These well-differentiated clades indicate that, historically, X. fastidiosa has been a clonal organism. Based on their synonymous-site divergence ( approximately 3%), these three clades probably originated more than 15,000 years ago, long before the introduction of the nonnative plants that characterize most infections. The sister clades of X. fastidiosa subsp. sandyi and X. fastidiosa subsp. piercei have synonymous-site evolutionary rates 2.9 times faster than X. fastidiosa subsp. multiplex, possibly due to generation time differences. Within X. fastidiosa subsp. multiplex, a low level ( approximately 0.1%) of genetic differentiation indicates the recent divergence of ALS isolates from the PP, PLS, and OAK isolates due to host plant adaptation and/or allopatry. The low level of variation within the X. fastidiosa subsp. piercei and X. fastidiosa subsp. sandyi clades, despite their antiquity, suggests strong selection, possibly driven by host plant adaptation.     

Impact of duration versus frequency of probing by Homalodisca vitripennis (Hemiptera: Cicadellidae) on inoculation of Xylella fastidiosa

Successful infection of the plant pathogenic bacterium Xylella fastidiosa (Wells) from an infected plant to a new host involves three main steps: 1) acquisition of the bacterium by a vector; 2) inoculation of a noninfected host plant by the vector; and 3) establishment of sufficient titers of X. fastidiosa in the host plant to sustain a chronic infection. Understanding the basic biology of the transmission process is a key to limiting the spread of plant diseases induced by X. fasdidiosa and reducing agricultural losses, especially those experienced in California since the introduction of a new vector, Homalodisca vitripennis (Germar) (Hemiptera, Cicadellidae) (formerly H. coagulata Say), the glassy-winged sharpshooter. In this study, H. vitripennis adults that acquired X. fastidiosa were allowed access to chrysanthemum plant cuttings for 30, 60, 90, or 120 min. The numbers of X. fastidiosa acquired (i.e., cells present in the insect foregut) and the number inoculated to the plant cuttings were separately determined using quantitative real-time polymerase chain reaction (PCR). In addition, the number of times glassy-winged sharpshooter stylets probed plant cuttings and the amount of time glassy-winged sharpshooter spent actively ingesting were monitored using video surveillance. Linear regression did not indicate a relationship between the number of X. fastidiosa cells inoculated into the plant cutting and either the titer of pathogen present in the insect or amount of time spent ingesting per probe. However, the number of probes significantly influenced the number of X. fastidiosa cells inoculated. Due to the highly variable nature of transmission, our model could not account for all observed variation as indicated by low R2 values. However, our results suggest that the mechanism of transmission is dependent on probing behaviors more than ingestion duration.     

Phenology and demography of Homalodisca coagulata (Hemiptera: Cicadellidae) in southern California citrus and implications for management

Populations of Homalodisca coagulata (Say) were sampled from citrus orchards in southern California, USA to characterize and quantify seasonal occurrences of nymphs and adults with the goal of identifying management opportunities through well-timed treatments and/or natural enemy releases. Higher densities of H. coagulata in 2001 contributed to a complete seasonal profile that began in early spring with the emergence of first instar nymphs and their progression through five nymphal instars lasting until mid-August. Adult emergence began in mid-June with peak adult densities attained from mid to late August followed by a gradual decline through autumn. A persistent and significant male bias was observed in the adult sex ratio from the time of first emergence through mid-October in oranges; the same trend was present in lemons, but with more variability. Adult densities gradually declined through the winter months into the following spring before rapidly increasing again in June as the 2002 spring generation of nymphs began emerging as adults. The seasonal timing of nymphs and adults in 2002 was nearly identical to that observed the previous year. Phenology data from both years were incorporated into a stochastic, temperature-dependent model that predicts the occurrences of H. coagulata stages through time. Applications of imidacloprid early in the spring generation of nymphs proved very effective at reducing nymphs and sustaining lower densities of adults through summer.     

Distribution and management of citrus in California: implications for management of glassy-winged sharpshooter

The epidemiology of Pierce's disease of grape (Vitis spp.) in California has changed over the past 10 yr due to the introduction of an exotic vector, Homalodisca vitripennis (Germar), the glassy-winged sharpshooter. Although this insect is highly polyphagous, citrus (Citrus spp.) is considered a preferred host and proximity to citrus has been implicated as a significant risk factor in recent epidemics of Pierce's disease in southern California. Consequently, a detailed knowledge of the distribution and management of citrus in relation to grape is needed to improve insect and disease management. Analysis of data on the area planted to these two commodities indicates that only five counties in California concomitantly grow >1,000 ha of grape and >1,000 ha of citrus: Riverside, Kern, Tulare, Fresno, and Madera counties. Comparison of the distribution of grape and citrus within each of these counties indicates that the percentage of grape that is in proximity to citrus is greatest for Riverside County, but the total area of grape that is in proximity to citrus is greater for Fresno, Kern, and Tulare counties. The use of carbamates, neonicotinoids, organophosphates, and pyrethroids as part of the citrus pest management program for control of key insect pests was compared among the same five counties plus Ventura County from 1995 to 2006. Ventura County was included in this analysis as this county grows >10,000 ha of citrus and has established glassy-winged sharpshooter populations. The use of these broad-spectrum insecticides was lowest in Riverside and Ventura counties compared with the other four counties. Analysis of historical trapping data at the county scale indicates a negative association of broad-spectrum insecticide use with glassy-winged sharpshooter abundance. These results are used to retrospectively analyze the Pierce's disease outbreaks in Kern and Riverside counties.