ABSTRACT
The vine mealybug, Planococcus ficus, is a significant pest of vineyards in all major grape growing regions of the world. This pest causes significant aesthetic damage to berry clusters through its feeding behavior and secretion of "honeydew", which leads to significant decreases in crop marketability. More importantly, the vine mealybug is a vector of several grapevine viruses which are the causal agent of grapevine leafroll disease, one of the most destructive and economically devastating diseases of the grape industry worldwide. As there is no cure for grapevine leafroll disease, the only control measures available to reduce its spread are to remove infected vines whilst simultaneously controlling mealybug populations. Using transcriptomic libraries prepared from male and female mealybugs and a draft genome, we identified and evaluated expression levels of members of the odorant receptor gene family. Interestingly, of the 50 odorant receptors identified from these P. ficus genetic resources, only 23 were found to be expressed in females, suggesting this flightless life stage has a decreased reliance on the olfactory system. In contrast, 46 odorant receptors were found to be expressed in the alate male life stage. Heterologous expression of eight of these receptors, along with the obligate co-receptor, Orco, in HEK293 cells allowed for the identification of two receptors that respond to lavandulyl senecioate, the sole constituent of the sex pheromone used by this species. Interestingly, one of these receptors, PficOR8, also responded to the sex pheromone used by the Japanese mealybug, Planococcus kraunhiae. The data presented here represent the first report of odorant receptor gene family expression levels, as well as the identification of the first sex pheromone receptor, in soft-scale insects. The identification of a receptor for the vine mealybug sex pheromone will allow for the development of novel, species-specific pest control tools and monitoring devices.
ABSTRACT
Risk assessments of chemical pesticides toward natural enemies are crucial for ensuring sustainable grapevine-integrated pest management. In this context, laboratory experiments were conducted to evaluate the toxicity of four insecticides (lambda-cyhalothrin, flupyradifurone, acetamiprid, and cyantraniliprole) and one fungicide (spiroxamine) commonly applied in German (European) vineyards on the pupae and adults of both Anagyrus vladimiri, a parasitoid of the vine mealybug Planococcus ficus, and Trichogramma evanescens, a parasitoid of the European grapevine moth, Lobesia botrana. The tested pesticides did not significantly affect the development of the pupal stage inside mealybug mummies or the emergence of the parasitoid A. vladimiri. The pesticides flupyradifurone, acetamiprid, and spiroxamine resulted in the highest mortality percentages for all emerged A. vladimiri parasitoids at 8 and 10 days after treatment compared with either in lambda-cyhalothrin or cyantraniliprole. However, all pesticides, except the diamide insecticide cyantraniliprole, significantly affected the development of the pupal stage and the emergence of the parasitoid T. evanescens. The percentages of T. evanescens emergence following the application of the fungicide spiroxamine or either lambda-cyhalothrin or flupyradifurone were significantly higher than those observed in the acetamiprid treatment. Regarding direct contact toxicity, the highest percentages (100%) of A. vladimiri adult parasitoid mortality were obtained in the flupyradifurone, acetamiprid, and spiroxamine treatments, while the lowest mortality percentages were observed in lambda-cyhalothrin, cyantraniliprole, and untreated control treatments. According to the IOBC classes of toxicity, flupyradifurone, acetamiprid, and spiroxamine were classified as harmful, while both lambda-cyhalothrin and cyantraniliprole were classified as slightly harmful to A. vladimiri adults. As such, all pesticides had a significant impact on the survival of exposed T. evanescens adults. The highest percentages of adult T. evanescens mortality were obtained in the flupyradifurone, acetamiprid, and spiroxamine treatments, with the fungicide spiroxamine resulting in significantly higher mortality percentages than either flupyradifurone or acetamiprid, while the lowest mortality percentages were found in the lambda-cyhalothrin and cyantraniliprole treatments. Therefore, applying the insecticides acetamiprid and/or flupyradifurone and the fungicide spiroxamine should be avoided when A. vladimiri and/or T. evanescens are naturally present or released in grapes. The insights gained from these two easy-to-rear parasitoid species allow analogous conclusions to be drawn for closely related species in vineyards belonging to either family Encyrtidae or Trichogrammatidae, which are not easy to rear. Interestingly, using the safer insecticides lambda-cyhalothrin and/or cyantraniliprole could be compatible with both parasitoid species, which could be sustainably exploited in either conservation or augmentative biological control in vineyards.
ABSTRACT
A survey of recently discovered vitiviruses was performed on 113 Croatian autochthonous grapevine cultivars from the national collection "Jazbina" using one-step RT-PCR. The presence of grapevine virus H (GVH) was confirmed in nine (7.9%) cultivars and grapevine virus G in eight (7.1%), while the presence of grapevine viruses I and J were not detected. GVH was transmitted by the vine mealybug (Planococcus ficus) from a source plant to grapevine seedlings with a 10.5% transmission rate using a combination of 10 first and second instars per plant with 48 and 72 h of acquisition and inoculation access period, respectively. Transmission correlated with the presence of grapevine leafroll-associated virus 3 (GLRaV-3) in the GVH-source plant and recipient seedlings. No alternative GVH host was identified. A comparison of 356 nt fragments of the RdRP and CP coding regions showed nucleotide identity between the Croatian GVH isolates in the range of 95.5-99.2% and 97.5-99.4% and amino acid identity between 95.8 and 100% and between 98.3 and 100%, respectively. Comparison with foreign isolates revealed nucleotide sequence similarity in the RdRP and CP between 94 and 100% and between 97.7-100%, respectively. To the best of our knowledge, this is the first report of GVH in Croatia and the first identification of the vine mealybug as a vector of GVH.
ABSTRACT
Grapevine leafroll disease is one of the most important virus diseases of grapevines and occurs in every major grape-growing region of the world. The vector-transmission mechanisms of the causative agent, Grapevine leafroll-associated virus 3 (GLRaV-3), remain poorly understood. We show that the vine mealybug, Planococcus ficus, feeds through a membrane feeding system on GLRaV-3 viral purifications from both V. vinifera and N. benthamiana and transmits the virus to test plants from plants from both species. Building on this strategy, we used an immunofluorescence approach to localize virions to two retention sites in P. ficus mouthparts. Assays testing molecules capable of blocking virus transmission demonstrated that GLRaV-3-transmission by P. ficus could be disrupted. Our results indicate that our membrane feeding system and transmission-blocking assays are a valid approach and can be used to screen other candidate blocking molecules.