Dyella-Like Bacterium Isolated from an Insect as a Potential Biocontrol Agent Against Grapevine Yellows.

Yellows diseases, caused by phytopathogenic bacteria of the genus Phytoplasma, are a major threat to grapevines worldwide. Because conventional applications against this pathogen are inefficient and disease management is highly challenging, the use of beneficial bacteria has been suggested as a biocontrol solution. A Dyella-like bacterium (DLB), isolated from the Israeli insect vector of grapevine yellows (Hyalesthes obsoletus), was suggested to be an endophyte. To test this hypothesis, the bacterium was introduced by spraying the plant leaves, and it had no apparent phytotoxicity to grapevine. Fluorescent in situ hybridization analysis showed that DLB is colonizing grapevine phloem. Because phytoplasmas inhabit the same niche, DLB interactions with this phytopathogen were examined. When the isolate was introduced to phytoplasma-infected Chardonnay plantlets, morphological disease symptoms were markedly reduced. The mode of DLB action was then tested using bioinformatics and system biology tools. DLB genome analysis suggested that the ability to reduce phytoplasma symptoms is related to inhibition of the pathogenic bacterium. These results provide the first step in examining the potential of DLB as a biological control agent against phytoplasmas in grapevine and, possibly, other agricultural crops.

Introduction of a putative biocontrol agent into a range of phytoplasma- and liberibacter-susceptible crop plants.

BACKGROUND: Phytoplasma, the causative agent of Bois Noir disease of grapevines, are vectored by the planthopper Hyalesthes obsoletus (Hemiptera: Cixiidae). A Dyella-like bacterium (DLB) isolated from H. obsoletus inhibits the growth of Spiroplasma melliferum, a cultivable relative of phytoplasma. Additional evidence suggests that DLB can reduce the symptoms of yellows disease in grapevine plantlets. The aim of this study was to test whether DLB could colonize a range of phytoplasma- and liberibacter-sensitive crop plants, and thus assess its potential agricultural use. RESULTS: Vitex agnus-castus, the preferred host plant of H. obsoletus was found to be a natural host of DLB, which was successfully introduced into a range of crop plants belonging to seven families. The most effective DLB application method was foliar spraying. Microscopy observation revealed that DLB aggregated on the leaf surface and around the stomata, suggesting that this is its route of entry. DLB was also present in the vascular tissues of plants, indicating that it moved systemically through the plant. CONCLUSIONS: DLB is a potential biocontrol agent and its broad spectrum of host plants indicates the possibility of its future use against a range of diseases caused by phloem-limited bacteria. © 2017 Society of Chemical Industry.

Bacterial associates of Hyalesthes obsoletus (Hemiptera: Cixiidae), the insect vector of bois noir disease, with a focus on cultivable bacteria.

The planthopper Hyalesthes obsoletus (Hemiptera: Cixiidae) is an important vector of phytoplasma diseases in grapevine. In the current study, the bacterial community compositions of symbionts of this insect were examined. Two dominant bacterial lineages were identified by mass sequencing: the obligate symbiont Candidatus Sulcia, and a facultative symbiont that is closely related to Pectobacterium sp. and to BEV, a cultivable symbiont of another phytoplasma vector, the leafhopper Euscelidius variegatus. In addition, one bacterium was successfully isolated in this study - a member of the family Xanthomonadaceae that is most closely related to the genus Dyella. This Dyella-like bacterium (DLB) was detected by FISH analysis in H. obsoletus guts but not ovaries, and its prevalence in H. obsoletus increased during the fall, suggesting that it was acquired by the host through feeding. We found that DLB inhibits Spiroplasma melliferum, a cultivable relative of phytoplasma, suggesting that it is a potential candidate for biological control against phytoplasma in grapevines.

Female detection of the synthetic sex pheromone contributes to the efficacy of mating disruption of the European grapevine moth, Lobesia botrana.

BACKGROUND: Studies of the mechanisms by which mating-disruption techniques control insect pest populations have traditionally focused on the effects of the species-specific sex pheromone on the male moths, while neglecting possible direct effects of the pheromone on females. Here, the effects of exposure to synthetic species-specific sex-pheromone on Lobesia botrana (European grapevine moth) females were tested. RESULTS: Females in vineyards that were treated with mating-disruption pheromone burst into short bouts of flying more frequently, but called significantly less frequently than females in untreated plots. Reduced calling caused by exposure to the species-specific sex-pheromone may increase the age at which females mate and thereby reduce female fecundity. Females that called in a pheromone-saturated environment experienced a decrease in number of oviposited eggs. A further decrease in reproductive success may occur if females delay oviposition when exposed to access of the synthetic pheromone. CONCLUSIONS: In addition to reducing the ability of males to locate females, the mating-disruption technique can suppress pest numbers as a consequence of its direct effects on females. The two mechanisms probably act synergistically.

Fitness cost of pheromone production in signaling female moths.

A secondary sexual character may act as an honest signal of the quality of the individual if the trait bears a cost and if its expression is phenotypically condition dependent. The cost of increasing the trait should be tolerable for individuals in good condition but not for those in a poor condition. The trait thus provides an honest signal of quality that enables the receiver to choose higher quality mates. Evidence for sex pheromones, which play a major role in shaping sexual evolution, inflicting a signaling cost is scarce. Here, we demonstrate that the amount of the major component of the pheromone in glands of Lobesia botrana (Lepidoptera) females at signaling time was significantly greater in large than in small females, that male moths preferred larger females as mates when responding to volatile signals, and small virgin females, but not large ones, exposed to conspecific pheromone, produced, when mated, significantly fewer eggs than nonexposed females. The latter indicates a condition-dependent cost of signaling. These results are in accordance with the predictions of condition-dependent honest signals. We therefore suggest that female signaling for males using sex pheromones bears a cost and thus calling may serve as honest advertisement for female quality.

Pest management programmes in vineyards using male mating disruption.

Israeli vine growers have been reluctant to adopt the mating disruption technique for control of the European vine moth, Lobesia botrana Den. & Schiff. Since the chemically controlled honeydew moth, Cryptoblabes gnidiella Mill., coexists with the European vine moth, growers have maintained that the use of mating disruption would fail to bring about a significant reduction in pesticide use. In this study, the efficacy of mating disruption techniques against C. gnidiella was tested, as well as the effect of these methods on pesticide use and damage to clusters when the method was employed against both of the pests in wine grapes. Comparisons were made between plots treated with (1) L. botrana mating disruption pheromone, (2) L. botrana and C. gnidiella mating disruption pheromones and (3) control plots. A significant difference in the number of clusters infested with the developmental stages of the moths was seen between pheromone-treated plots and controls, while no such difference was observed between plots treated with one versus two pheromones. A similar pattern was observed in the number of insecticide applications; the greatest number of applications was used in control plots, followed by plots treated with L. botrana mating disruption pheromone and by plots treated with pheromones against both pests, in which no pesticides were applied.

Vitex agnus-castus is a preferred host plant for Hyalesthes obsoletus.

Hyalesthes obsoletus Signoret (Homoptera: Cixiidae) is a polyphagous planthopper that transmits stolbur phytoplasma (a causative agent of "yellows" disease) to various weeds, members of the Solanaceae, and wine grapes (Vitis vinifera L.) in Europe and the Middle East. Planthoppers were collected by hand vacuuming eight native plant species. Vitex agnus-castus L., a shrub in the Verbenaceae, hosted the largest number of H. obsoletus, although Olea europaea L. also served as a host for adults. Using a Y-olfactometer, we compared the planthoppers relative preference for V. agnus-castus, Convolvulus arvensis, and V. vinifera. V. agnus-castus was more attractive to both male and female H. obsoletus than the other plants. H. obsoletus antennal response was stronger to volatiles collected from V. agnuscastus than from Cabernet Sauvignon variety of V. vinifera. To determine if V. agnus-castus would serve as a reservoir for the pathogen, H. obsoletus were collected from leaf and stem samples of native V. agnus-castus, and were tested by polymerase chain reaction (PCR) for the presence of phytoplasma DNA. While 14% and 25% (2003 and 2004, respectively) of the insects tested positive for phytoplasma DNA, none of the plant samples tested positive. To determine if V. agnus-castus could serve as a host plant for the development of the planthopper, we placed emergence cages beneath field shrubs and enclosed wild-caught H. obsoletus in a cage with a potted young shrub. We found adult H. obsoletus in the emergence cases and planthopper nymphs in the soil of the potted plant. We concluded that V. agnus-castus is attractive to H. obsoletus, which seems to be refractory to phytoplasma infections and warrants further testing as a trap plant near vineyards.

Mating disruption of Lobesia botrana (Lepidoptera: Tortricidae): effect of pheromone formulations and concentrations.

The reluctance of Israeli vine growers to adopt the mating disruption technique to control the moth Lobesia botrana Den. & Schiff. has been attributed to the high cost of this method compared with that of traditional insecticide control. In this study, we tested the possibility of reducing the cost, first by testing different pheromone formulations (and thus open the market for competition) and second by reducing the pheromone concentration used in vineyards. Comparisons were made between two pheromone formulations--Shin-Etsu (Tokyo, Japan) at 165 g/ha and Concep (Sutera, Bend, OR) at 150 g/ha--and between two concentrations of Shin-Etsu, 165 and 110 g/ha. Pheromone dispensers were placed at the onset of the second moth generation. Comparison of the numbers of clusters infested with eggs and larvae of L. botrana showed no significant differences in the performance, either between the two formulations, or between the two tested concentrations. The results suggest that 1) the two formulations are equally effective, and 2) a low pheromone concentration is sufficient to maintain good control of small populations of L. botrana. However, when the population is high, pest control efficacy is not improved by increasing the pheromone concentration. Therefore, in the interest of reducing the relatively high cost of mating disruption, we emphasize that increasing the pheromone concentration does not provide improved control of high populations of L. botrana. The cost of mating disruption can be diminished by reducing the applied pheromone concentration and by using the least expensive pheromone formulations