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Finding a partner to mate with may be only part of ensuring successful siring of offspring. Females often exhibit cryptic female choice (CFC) during or after copulation, which can influence whose sperm from her multiple partners is chosen for egg fertilization. Known behavioral mechanisms for CFC include assessment of males by their nuptial gifts, duration of copulation, and seminal fluid contents. In this study, the glassy-winged sharpshooter, Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae), behaviors during the course of copulation were investigated. Glassy-winged sharpshooter (GWSS) use vibrational communication before copulation occurs. However, little is known about behaviors that occur during and after copulation. Results from this study determined that vibrational signaling also occurs during copulation. Vibrational signals similar to those emitted during precopulatory communication were identified during copulation alongside a new, ‘hum-like’ signal that typically occurred within 10 s after the pair joined in copulation. In addition, results determined the duration of copulation was on average of 15 h, though with a 10-h range (8.5–18.5 h) among observed male–female pairs. Finally, both males and females mated more than once. Collectively, results identified key reproductive parameters required for CFC to occur in GWSS. The study expands on the known animals that use CFC and emphasizes the role that copulatory vibrational communication may play setting the foundations for future more in-depth studies. Understanding of insect behaviors necessary for successful production of offspring is important from an ecological perspective and for development of pest control methods.

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.

Vertical transmission of Homalodisca vitripennis reovirus (HoVRV) from glassy-winged sharpshooter (GWSS, Homalodisca vitripennis (Germar)) females to progeny occurred in laboratory assays at frequencies too low (2.6%–15.4%) to account for HoVRV incidence (90–100%) in field populations resident in citrus. Because citrus is immune to HoVRV and no plant host is known, horizontal transmission of HoVRV from insect-to-insect was evaluated. Exposure of colony-reared, virus-free test nymphs to HoVRV-infected source adults held in the same cage for 10 days on virus-immune cowpea resulted in HoVRV transmission (13.3%–30.7%) to test nymphs. HoVRV was not transmitted when exposure was indirect and required passive movement of virions through the xylem of immune citrus seedlings. Collectively, these results demonstrate direct insect-to-insect horizontal transmission of HoVRV, providing a plausible explanation for high incidence of HoVRV in GWSS field populations in the absence of efficient vertical transmission or a plant host.

BACKGROUND Glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae) is an important vector of the bacterium Xylella fastidiosa, the causal agent of Pierce's disease of grapevine. Area-wide insecticide applications have suppressed GWSS populations for ∼ 25 years, but reduced levels of insecticide susceptibility have been reported. Therefore, alternative methods of control are needed. Objectives of this study were to evaluate the efficacy of playback of vibrational mating communication signals for disrupting mating of GWSS in a natural vineyard setting and to evaluate spectral properties of signal transmission through vineyard trellis. RESULTS Playback reduced mating of GWSS on grapevines. A total of 28 (of 134) male-female pairs mated in the control treatment (silence) and only one (of 134) pair mated when treated with the vibrational signal playback. Playback of vibrational signals through vineyard trellis was affected by distance from the signal source, with frequency composition being the highest at the source and lowest on vines positioned away from the source. Frequency composition in canes housing test insects decreased exponentially as distance from the source increased, whereas the relative amplitude of analyzed frequencies decreased linearly. CONCLUSION Although further studies are needed prior to method implementation, data from this study continue to support integration of vibrational mating disruption with current methods to suppress GWSS populations. Published 2018. This article is a U.S. Government work and is in the public domain in the USA.

BACKGROUND The glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, is an important pest of grapevines due to its ability to transmit Xylella fastidiosa, the causal agent of Pierce's disease. GWSS mating communication is based on vibrational signals; therefore, vibrational mating disruption could be an alternative to insecticides for suppression of GWSS population. Our objectives were to identify spectral features of female signal that elicit male signaling, design disruptive signals able to alter male perception and acceptance of a female, and determine the signal intensity required for future field applications. RESULTS Results showed that male responses to playback of modified female signals were significantly reduced by 60-75%when part of the female signal spectral components above or below 400 Hz were deleted. Playback bioassays showed that transmission of an 80 Hz pure frequency tone to plants completely suppressed male signaling to female signal playback, even if the disruptive signal amplitude was 10 dB lower than the female signal playback. CONCLUSION Although the mechanism underlying cessation of male signaling activity in the presence of disruption is not yet understood, results suggest that an 80 Hz vibrational signal should be tested in laboratory and field experiments to assess its efficacy in disrupting mating of GWSS.

The glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, is an important vector of Xylella fastidiosa, the causal agent of Pierce’s disease of grapevine. GWSS control relies mainly on insecticides; therefore, an alternative method, such as vibrational mating disruption, is required. However, knowledge of GWSS intraspecific communication is necessary to evaluate applicability of such methods. Mating behavior and associated vibrational signals were described in different social contexts: individuals, pairs, and one female with two competing males. Behavioral analysis showed that GWSS mating communication involved the emission of three male and two female signals, with specific roles in two distinct phases of mating behavior, identification and courtship. Mating success depended on vibrational duets between genders, which were temporarily interrupted in the presence of male rivalry. Male rivalry behavior involved the emission of three distinct rivalry signals. Two rivalry signals resemble female signals and were associated with replacement of the female in the duet by the rival male. The third rivalry signal was emitted by competing males. Data suggested that rival males used mimicry and hostile signals to interrupt the ongoing duet and gain access to a female. In the future, knowledge acquired from this study will be essential to develop a mechanical mating disruption method for GWSS control.

Xylella fastidiosa (Wells) is a xylem-limited bacterium that causes Pierce’s disease of grapevines. The bacterium is transmitted by insect vectors such as the glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar). Experiments were conducted to compare the role of selected X. fastidiosa genes on 1) bacterial acquisition and retention in GWSS foreguts, and 2) transmission to grapevines by GWSS. Bacterial genotypes used were: mutants Xf-ΔpilG, Xf-ΔpilH, Xf-ΔgacA, and Xf-ΔpopP; plus wild type (WT) as control. Results showed that Xf-ΔpilG had enhanced colonization rate and larger numbers in GWSS compared with WT. Yet, Xf-ΔpilG exhibited the same transmission efficiency as WT. The Xf-ΔpilH exhibited poor acquisition and retention. Although initial adhesion, multiplication, and retention of Xf-ΔpilH in GWSS were almost eliminated compared with WT, the mutation did not reduce transmission success in grapevines. Overall, Xf-ΔgacA showed colonization rates and numbers in foreguts similar to WT. The Xf-ΔgacA mutation did not affect initial adhesion, multiplication, and long-term retention compared with WT, and was not significantly diminished in transmission efficiency. In contrast, numbers of Xf-ΔpopP were variable over time, displaying greatest fluctuation from highest to lowest levels. Thus, Xf-ΔpopP had a strong, negative effect on initial adhesion, but adhered and slowly multiplied in the foregut. Again, transmission was not diminished compared to WT. Despite reductions in acquisition and retention by GWSS, transmission efficiency of genotypes to grapevines was not affected. Therefore, in order to stop the spread of X. fastidiosa by GWSS using gene-level targets, complete disruption of bacterial colonization mechanisms is required.

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.

The glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, is a generalist insect and a major pest of grapevines. GWSS is a vector of the bacterium Xylella fastidiosa, causal agent of Pierce’s disease that can lead to grapevine death within few years after infection. This chapter discusses the problem and current GWSS control methods. Then, the focus changes to efforts in developing a novel control method that uses synthetic vibrational signals to disrupt mating and, thus, population growth. A step-by-step method for creating effective playback signals is described and discussed. The method was termed “D.I.E.,” which stands for Describe, Identify, and Execute. The first step is to describe the basic biology of the insect pest with emphasis on communication behaviors. From there, one can begin identifying which candidate signals disrupt communication. Finally, execution tests are conducted to determine which signals effectively disrupt mating in laboratory, and more importantly, in the field. While there are still steps needed for large-scale implementation in the field, the basic biological questions related to whether synthetic vibrational signals can disrupt mating are answered affirmatively. The next direction will be to develop a mechanism of signal transmission across large areas. In addition, season long field trials are needed, taking into consideration natural insect movements into and out of treated areas.

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.

Glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar), is a vector of the xylem-inhabitant bacterium Xylella fastidiosa Wells et al., which causes Pierce’s disease of grapevines. Current GWSS control strategies in California, USA include area-wide insecticide applications and mass release of mymarid egg parasitoids, including Gonatocerus ashmeadi Girault. Gas chromatography–mass spectrometry was used to identify ( E )-β-ocimene and ( E , E )-α-farnesene as volatiles emitted from grapevines on which GWSS had previously fed and oviposited. Attractiveness of female G. ashmeadi to sugar-based formulations containing either ( E )-β-ocimene, ( E , E )-α-farnesene, or a mixture of both was evaluated using Y-tube olfactometry. When exposed to synthetic formulation containing a mixture of ( E )-β-ocimene and ( E , E )-α-farnesene vs. blank control, 61% of G. ashmeadi females initially chose the synthetic formulation. After the initial choice for a Y-tube arm, females visited the Y-tube arm connected to the source of formulation more often than it did to the arm connected to a blank control. There was no difference in the female’s time spent in the arm connected to the formulation. When testing formulations containing either ( E )-β-ocimene or ( E , E )-α-farnesene alone, there was a 1:1 ratio between the proportion of parasitoid’s first choice, visits, and residence time. Results suggest that synthetic formulations containing mixtures of certain plant volatiles may be used to localize GWSS egg parasitoids in vineyard systems. Results are discussed in the context of potential applications in GWSS biological control programs.

Gonatocerus morrilli (Howard) (Hymenoptera: Mymaridae) is an egg parasitoid used in California, USA to control glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae). Virgin GWSS females deposit non-fertilized eggs and mated females can exhaust sperm reserves for egg fertilization. However, nothing is known about Gonatocerus spp. performance when using non-fertilized GWSS eggs as hosts. Host age preference for oviposition and suitability of non-fertilized GWSS eggs as hosts for G. morrilli reproduction were investigated to determine whether non-fertilized eggs on sentinel plants could be used to monitor egg parasitoid populations. Gonatocerus morrilli parasitized all ages of GWSS eggs (1–8 days old) regardless if the host egg was fertilized or not. In choice tests (fertilized versus non-fertilized eggs), parasitoids failed to emerge as adults from non-fertilized eggs more often than from fertilized eggs. The results indicate that non-fertilized eggs were accepted by G. morrilli as suitable hosts for oviposition, but were less suitable for immature development compared to fertilized eggs.

The glassy-winged sharpshooter, Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae), is a xylem fluid-ingesting leafhopper that transmits χylella fastidiosa Wells et al., a plant-infecting bacterium that causes several plant diseases in the Americas. Although the role of plant water stress on the population density and dispersal of H. vitripennis has been studied, nothing isknown about the effects of plant water stress on the transmission of χ. fastidiosa by H. vitripennis.Alaboratory study was conducted to determine the influence of plant water stress on the sharpshooter stylet probing behaviors associated with the acquisition and inoculation of χ. fastidiosa. Electrical penetration graph was used to monitor H. vitripennis feeding behaviors for 20-h periods on citrus [Citrus sinensis (L.) Osbeck] and almond [Prunus dulcis (Miller) D.A. Webb] plants subjected to levels of water stress. Adult H. vitripennis successfully located xylem vessels, then performed behaviors related to the evaluation of the xylem cell and fluid, and finally ingested xylem fluid from citrus and almond plants under the tested fluid tensions ranging from -5.5 to -33.0 bars and -6.0 to -24.5 bars, respectively. In general, long and frequent feeding events associated with the acquisition and inoculation of χ. fastidiosa were observed only in fully irrigated plants (i.e., >-10 bars), which suggests that even low levels of plant water stress may reduce the spread of χ. fastidiosa. Results provided insights to disease epidemiology and support the hypothesis that application of regulated deficit irrigation has the potential to reduce the incidence of diseases caused by χ. fastidiosa by reducing the number of vectors and by decreasing pathogen transmission efficiency.

A two-year study was conducted in a citrus orchard, Citrus sinensis L., to determine frequency of predation on glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar). A total of 1,578 arthropod predators, representing 18 taxa, were collected and assayed for the presence of GWSS egg protein by an enzyme-linked immunosorbent assay using a Homalodisca-species and egg-specific monoclonal antibody and then by polymerase chain reaction using a H. vitripennis-specific DNA marker. The gut content analyses revealed the presence of GWSS remains in the gut of 2.28 % of the total arthropod predator population, with 3.09 % of the spiders and 0.59 % of the insect predators testing positive. Moreover, a comparison of the two assays indicated that they were not equally effective at detecting GWSS remains in predator guts. Low frequencies of GWSS detection in the gut of predators indicated that GWSS are not a primary prey and that predators may contribute little to suppression of this pest in citrus.

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 quanti...

The agricultural pest, glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, relies primarily on successful vibrational communication across its home plant. Males and females engage in a vibrational duet to identify correct species, attractiveness of mate, and location on the plant. The signal produced by these animals has a dominant frequency component between 80 and 120 Hz, with harmonics spaced approximately 100 Hz apart. However, our analysis revealed that not all harmonics are present in every recorded signal. Therefore, we sought to understand how the GWSS vibrational communication signal changes over distance on the plant. We have confirmed that first, with increasing distance fewer high frequency harmonics are present. Second, at distances of only 50 cm, there is a difference in the latency of signal arrival based on the frequency, with higher frequencies arriving sooner. Finally, the animal appears to generate no airborne signal component, yet, the low frequencies are clearly detectable in neighboring plants by the signal “jumping” from leaf-to-air-to-leaf. Together, these results highlight the complexity of vibration transmission in plants and the possibility of alteration and disruption of the GWSS signal.

Anagrus epos Girault (Hymenoptera: Mymaridae) is a natural enemy candidate for a classical biological control program targeting the glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae), in California. Little is known about the biology or ecology of A. epos when it utilizes GWSS eggs as a host. Here, we report the results of laboratory studies that describe the host age preference for oviposition, longevity of A. epos adults provided with different food sources, and developmental rates at six different constant temperature regimes. Anagrus epos is a gregarious parasitoid in GWSS eggs with up to 14 adults emerging from each GWSS egg. In choice and no-choice tests for oviposition, A. epos females successfully parasitized all developmental ages of GWSS eggs (1–8 days old). In choice tests, parasitism rates were significantly higher in 1-, 3-, 4-, and 5-day-old GWSS eggs than in 2-, 6-, 7-, and 8-day-old eggs. If provided with honey and water, honey only, water only, or no food or water, A. epos females lived on average 8.2, 4.7, 2.6, and 1.6 days, respectively. Anagrus epos required 294.1 degree-days above a lower temperature threshold of 12.4 °C to develop from egg to adult (eclosion). Our results provide baseline information useful in the development of an efficient parasitoid mass rearing program for A. epos release and evaluation in California.

Gonatocerus ashmeadi Girault is an egg parasitoid with potential use in glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar), biological control in California. We tested the hypotheses that (1) G. ashmeadi uses chemical volatiles as cues to find hosts, (2) its success in host finding varies among plant species, and (3) the release of semiochemicals is systemically induced by GWSS feeding and oviposition. Using a glass Y-tube olfactometer, we determined G. ashmeadi’s preference for volatiles of infested versus uninfested lemon, grapevine, red-tip photinia, and crape myrtle plants. The parasitoid’s first choice to, its residence time, and the number of visits per Y-tube arm were used to assess its plant choice. G. ashmeadi chose the infested lemon, red-tip photinia, or grapevine plant significantly more often (62%, 67%, and 63%, respectively) than uninfested hosts, whereas they chose infested and uninfested crape myrtle equally often (49%). Females spent more time in and made more visits to the olfactometer arm connected to the infested plant than to the arm connected to the uninfested plant, except when testing crape myrtle. When testing for a systemic plant response to GWSS infestation, only infested lemon was significantly more attractive to the parasitoids than was an uninfested plant. These results suggest that G. ashmeadi females use chemical volatiles as cues to find GWSS egg masses and that host finding success varies among GWSS host plants. Some plants used by GWSS as oviposition hosts may serve as an escape from parasitization by G. ashmeadi and act as reservoirs for new GWSS individuals.

A new virus species of the genus Phytoreovirus was isolated from glassy-winged sharpshooter (GWSS), Homalodisca vitripennis Germar (Hemiptera: Cicadellidae), in California and designated here as Homalodisca vitripennis reovirus (HoVRV). Extraction of nucleic acid from GWSS adults collected from three Californian populations revealed an array of double-stranded (ds) RNA species that was soluble in 2 M LiCl and resistant to degradation upon exposure to S1 nuclease and DNase. Analysis of nucleic acid samples from single GWSS adults indicated that HoVRV dsRNA accumulated to high titer in individual insects. Double-shelled isometric virus particles purified from GWSS adults resembled those observed in thin sections of GWSS salivary glands by transmission electron microscopy. Purified HoVRV virions contained 12 dsRNA segments that, based on complete nucleotide sequences, ranged in size from 4475 to 1040 bp. Sequence comparisons indicated that the HoVRV dsRNA segments were most closely related (58.5 to 43.7% nt sequence identity) to the corresponding genome segments of Rice dwarf virus (RDV). Each HoVRV dsRNA segment encoded a single open reading frame (>300 nts) except for segment 11, which appears to be dicistronic. Terminal nucleotide sequences of HoVRV positive-sense RNAs were similar to other phytoreoviruses (GGCG or GGCA at the 5′-end and UGAU or CGAU at the 3′-end) with adjacent imperfect inverted repeats potentially able to base pair. Phylogenetic analyses of the RNA-directed RNA polymerase (encoded by segment 1) and the outer capsid protein (encoded by segment 8) confirmed placement of HoVRV as a species of the genus Phytoreovirus sharing a most recent common ancestor with RDV. Reverse transcription-polymerase chain reaction assays revealed that HoVRV infection of GWSS in California was common and that the virus also occurred in GWSS populations from the Carolinas and Texas.