RESUMO
"Flavescence dorée" (FD)-related phytoplasmas are widespread in alder in Germany and their transmission to grapevine represents a high risk for FD outbreaks when the primary vector, Scaphoideus titanus, becomes present in the future. Therefore, the potential role of the Deltocephalinae leafhopper species in transmitting FD-related phytoplasmas from alder to grapevine was studied in extensive transmission trials conducted between 2017 and 2020. The transmission capacity of autochthonous Allygus spp. and the invasive Orientus ishidae captured on infected alder trees was tested under controlled conditions using various test designs, including grouped insects and single-insect studies. The latter experiments were analyzed in terms of survival probability, transmission success and phytoplasma load in the insects, measured by quantitative PCR. A minimum inoculation titer (MIT) required for successful transmission to alder was defined for both Allygus spp. and O. ishidae. While Allygus spp. exhibited slightly better survival on Vitis vinifera compared to O. ishidae, the latter displayed higher phytoplasma loads and greater transmission success. Although all species were capable of infecting alder seedlings, O. ishidae was able to transmit 16SrV-phytoplasmas directly to single grapevines. Infective adults of O. ishidae were captured from the beginning of July until the end of August, while Allygus spp. were only considered infective towards the end of the season. Thus, O. ishidae likely poses a higher risk for FD transmission from alder to grapevine, albeit at a very low level, as only five out of 90 transmission trials to V. vinifera were successful.
RESUMO
Flavescence dorée (FD) and Bois noir (BN) are the principal grapevine yellows diseases in Europe caused by distinct phytoplasmas: FD by 16SrV phytoplasmas (FDp), BN by Candidatus Phytoplasma solani. FDp is spread epidemically by the introduced Nearctic Deltocephalinae Scaphoideus titanus and is listed as a quarantine pest in the European Union (Regulation (EU) 2019/2072). Black Alder (Alnus glutinosa) is a common asymptotic host of 16SrV phytoplasmas in Europe and considered the original host of FDp (Malembic-Maher et al. 2020). Palatinate grapevine yellows (PGY) transmitted from alder to grapevine by the Macropsinae Oncopsis alni (Maixner et al. 2000) is not transmissible by S. titanus, unlike isolates transmitted by the autochthonous Deltocephalinae Allygus spp. and the invasive Orientus ishidae (Malembic-Maher et al. 2020). Germany is considered free from FD in grapevine and from its vector. A single case in a nursery in 2014 was eradicated (EPPO 2017). Since S. titanus was detected in 2016 in the neighboring French Region of Alsace, monitoring of FD was carried out in Germany. It was focused on vineyards within a distance of 100 m from stands of alder trees. A geodata-based risk map (Jalke 2020) was used to identify those plots. All symptomatic vines sampled until September 2020 proved to be infected by BN or, occasionally, by PGY. Eight vines with typical symptoms were sampled in vineyards adjacent to alder stands in the winegrowing region of Rheinhessen in September 2020. Symptoms comprised leaf rolling and discoloration, incomplete lignification, black pustules on shoots, dried inflorescences and shriveled berries. Diseased shoots were black and necrotic in December. Leaf midribs were sampled for total DNA extraction. The phytoplasma 16S rRNA gene was amplified by generic primers R16F2/R2-mod followed by a nested PCR using 16Sr(V) group-specific primers R16(V)F1/R1, and primers R16(I)F1/R1 (Lee et al. 1995) to detect 'Candidatus Phytoplasma solani', associated with BN. While BN was detected in seven vines, one sample tested positive for 16SrV phytoplasma. This result was confirmed by triplex real-time Taq-Man assay based on rpl14 gene sequences (IPADLAB), by multiplex real-time PCR of map locus as well as by Loop-mediated isothermal amplification (LAMP) according to the EPPO diagnostic standard PM 7/079(2) (EPPO 2016). PCR-products of the map- and the vmpA-Gene (Malembic-Maher et al., 2020) were sequenced and compared to reference sequences to distinguish between FD- and non-FD genotypes. The isolate from the diseased vine exhibited 100% identity with map-M38 (Accession No. LT221933), a genotype of the map-FD2 cluster. The same genotype was detected in A. glutinosa and Allygus spp. sampled at the infested site. A 234 bp sequence of the first repeat of the vmpA-gene showed 100% identity with the S. titanus transmitted isolate FD-92 (Accession No. LN680870) of the vmpA-II cluster. It can be concluded, that the 16SrV-isolate detected in a symptomatic grapevine is infected by FD and not PGY. This is the first report of FD in a vineyard in Germany. The infected vine of cv. Silvaner was 25 years old. While infected planting material is an unlikely source of the infection, a transmission of FDp from alder is highly probable. Finding a single FD-infection after several years of testing implies a low risk originating from the wild compartment, but the approach of the vector S. titanus justifies further monitoring activities. The infected vine was eradicated.
RESUMO
The identification of volatile organic compounds (VOCs) leading to short-range attraction and oviposition of the European grapevine moth Lobesia botrana and European grape berry moth Eupoecilia ambiguella (Lepidoptera: Tortricidae) is crucial in order to establish bait-based decision support systems for control of these pests. Therefore, we developed a method to measure the real-time behavioral response of female moths to VOCs using a four-chamber olfactometer coupled with a video tracking system. Ten synthetic VOCs were selected for this study: (S)-(-)-perillaldehyde, (E)/(Z)-linalool oxide, (±)-limonene, linalool, (E)-ß-caryophyllene, α/ß-farnesene, (-)-α-cedrene, methyl salicylate and cumene. The effect of VOCs on egg deposition was determined using a dual-choice oviposition test, whereas perception by female antennae was verified using electroantennography (EAG). During video tracking, females responded to volatile compounds emitted by grapevine with higher antennae and ovipositor activity than to air control. (E)/(Z)-linalool oxide, cumene and (S)-(-)-perillaldehyde released ovipositor activity of L. botrana, while the latter provoked oviposition. (R)/(S)-limonene affected ovipositor activity of E. ambiguella, whereas none of the VOCs tested attracted for oviposition. The results suggest that females have the ability to perceive specific VOCs by the antennae but also by the ovipositor, which could attract or repel for egg deposition.
RESUMO
Greencover crops are widely recommended to provide predators and parasitoids with floral resources for improved pest control. We studied parasitism and predation of European grapevine moth (Lobesia botrana) eggs and pupae as well as predatory mite abundances in an experimental vineyard with either one or two sowings of greencover crops compared to spontaneous vegetation. The co-occurrence between greencover flowering time and parasitoid activity differed greatly between the two study years. Parasitism was much higher when flowering and parasitoid activity coincided. While egg predation was enhanced by greencover crops, there were no significant benefits of greencover crops on parasitism of L. botrana eggs or pupae. Predatory mites did not show an as strong increase on grapevines in greencover crop plots as egg predation. Overall, our study demonstrates only limited pest control benefits of greencover crops. Given the strong within- and between year variation in natural enemy activity, studies across multiple years will be necessary to adequately describe the role of greencover crops for pest management and to identify the main predators of L. botrana eggs.
