Bois noir phytoplasma (BNp), widespread in wine-producing areas of Europe and endemic in France and Italy, is classified in the 16SrXII-A subgroup, whose members are referred to as Stolbur phytoplasmas. The 16S rDNA gene of Stolbur phytoplasma shows low variability, and few non-ribosomal genes are available as markers to assess variation among isolates. We used the Stolbur-specific stol-1H10 gene, encoding a putative membrane-exposed protein, to investigate genetic diversity of French and Italian BNp isolates from plants and insects. Amplification of stol-1H10 from infected grapevines, weeds, and Hyalesthes obsoletus produced fragments of three sizes, and restriction fragment length polymorphism analysis divided these amplicons further into 12 profiles (V1 to V12). French BNp isolates were more variable than Italian ones, and different profiles were present in infected grapevines from France and Italy. Isolate V3, most abundant among Italian affected grapes but present among French ones, was found in one Urtica dioica sample and in all H. obsoletus collected on this species. Four Italian-specific profiles were represented among infected Convolvulus arvensis, the most frequent of which (V12) was also detected in H. obsoletus collected on this species. Most of the variability in the stol-1H10 sequence was associated with type II on the tuf gene.
Pheromone-mediated mating disruption (MD) is currently considered an effective strategy to control the European grapevine moth (EGVM), Lobesia botrana, with a successful interference on natural female calling during the male searching flight. However, little is known on the impact of the hour of the day on EGVM male flight. While various models forecasting the day of maximum presence of males per flight have been developed, field research on the male flight activity over the hours of the day is scarce. Hence, we used video camera-assisted pheromone traps to allow a continuous monitoring of EGVM flights over daylight and night hours, quantifying captures of males. Experiments were carried out in three vineyards located in northern Spain over two years (2016 and 2017). Results showed that EGVM flight mainly occurred between 21:00 and 23:00 hours. Furthermore, male catches significantly differed over the study year, annual flight period and vineyard. Most of the dispensers used worldwide for L. botrana MD continuously release the main sex pheromone component [(7E,9Z)-7,9-dodecadien-1-yl acetate], except for some automatic devices releasing puffs of sex pheromones at selected time intervals. The findings presented here can be useful to optimize the MD technique, identifying selected time intervals when the release of EGVM synthetic pheromones can be concentrated, boosting MD efficacy against this important pest, minimizing the release of synthetic sex pheromone molecules in the environment and reducing application costs.
Pheromone-mediated mating disruption (MD) is widely used as a control tool to manage the European grapevine moth (EGVM), Lobesia botrana. Most of the MD formulations are "passive" reservoir dispensers, which need to be used at a rather large number of units per hectare. A promising alternative is represented by automatic aerosol devices, releasing pheromone puffs at programmed time intervals. Herein, we investigated the effectiveness of MD aerosol product Isonet® L MisterX841 in reducing EGVM infestation on grape in comparison to the reference MD product Isonet® L and the grower's standard. Experiments were carried out over 2 years in two different study sites of Aragon region (Spain). EGVM male catches were monitored using traps baited with the female sex pheromone. The effectiveness of MD formulations against the three generations of EGVM was assessed by determining the percentage of infested bunches and the number of nests per bunch. As expected, a much greater amount of male catches in the grower's standard over Isonet® L MisterX841 and Isonet ® L was observed. No significant differences about EGVM male catches were found in vineyards where Isonet® L MisterX841 and Isonet® L were used. EGVM-infested bunches, as well as number of nests per bunch, were higher in the grower's standard, if compared to vineyards where we tested Isonet® L MisterX841 and Isonet® L. However, the employ of the latter led to a lower EGVM bunch infestation, if compared to Isonet® L MisterX841. Overall, the MD approach proposed here is effective against EGVM. These aerosol devices require a lower number of units per hectare if compared to hand-applied dispensers, saving labor costs and contributing to reduce plastic disposal in agricultural settings.
Laboratory trials were carried out on wild individuals of Reptalus quinquecostatus (Cixiidae), a potential vector of stolbur phytoplasma to grapevine, to assess its ability to inoculate the phytoplasma in artificial feeding medium. Seventyseven specimens of the cixiid were tested on a sucrose-TE (Tris-ethylenediaminetetraacetic acid) diet and 62 of them survived less than 24 h. Polymerase chain reaction (PCR) assays performed on the insect bodies detected the presence of stolbur phytoplasma, with an infection rate of 32.5%. Restriction fragment length polymorphism analysis of the tuf gene, amplified by PCR, revealed Vergilbungskrankheit type I (VK-I) in 20 specimens, VK-II in 4 specimens and both types in 1 specimen. Ten of the 25 infected R. quinquecostatus specimens successfully inoculated VK-I in the sucrose solution, that is, a 40% inoculation efficiency despite the brief survival. The results indicate that R. quinquecostatus is a competent species to transmit the stolbur phytoplasma in artificial conditions. The repeated observation of adults feeding on grapevine strengthens the hypothesis that the species is a vector of stolbur phytoplasma to this plant.
This chaper describes the pest significance in Italian vineyard, as well as the current management systems and approaches for the vine moth (Lobesia botrana) and their limitations. The implementation (including the development and implementation of education and technology transfer programmes); compatibility with crop management and control of co-occurring pests; efficacy in controlling the target pest(s); and costs and benefits of the AWPM programme are discussed. Future directions are given.
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