Influence of Vineyard Inter-Row Management on Grapevine Leafhoppers and Their Natural Enemies.

Inter-row management in vineyards can influence the abundance of grapevine pests and their natural enemies. In 2013-2015, in a vineyard in northeastern Italy, the influence of two vineyard inter-row management strategies (i.e., alternate mowing, AM, and periodical tillage, PT) on the population dynamics of grapevine leafhoppers Hebata vitis and Zygina rhamni and their natural enemies, the mymarid Anagrus atomus and spiders (Araneae), and other hymenopteran parasitoids, were studied with different survey approaches. The infestations of both leafhoppers were lower in AM than PT due to the reduced leafhopper oviposition and higher nymph mortality in AM. This occurred although leafhopper egg parasitization by A. atomus was greater in PT than AM according to a density-dependent relationship with the leafhopper egg amount. Hymenopteran parasitoids other than A. atomus were the most abundant in AM, probably due to the higher availability of nectar and pollen than in PM. The significantly higher population densities of hunting spiders in AM than PT can be associated with the higher predation of leafhopper nymphs. Therefore, the study demonstrated that the alternate mowing of vineyard inter-rows enhances the abundance of natural enemies, such as spiders and hymenopteran parasitoids, and can contribute to grapevine leafhopper pest control.

Efficacy of Conventional and Organic Insecticides against Scaphoideus titanus: Field and Semi-Field Trials.

Scaphoideus titanus is the main vector of phytoplasmas associated with Flavescence dorée (FD), one of the most serious threats to viticulture in many European countries. To minimize the spread of this disease, mandatory control measures against S. titanus were decided in Europe. In the 1990s, the repeated application of insecticides (mainly organophosphates) proved to be an effective measure to control the vector and the related disease in north-eastern Italy. These insecticides and most of the neonicotinoids were recently banned from European viticulture. Serious FD issues detected in the recent years in northern Italy could be related to the use of less effective insecticides. Trials aimed at evaluating the efficacy of the most used conventional and organic insecticides in the control of S. titanus have been performed in semi-field and field conditions to test this hypothesis. In efficacy trials, carried out in four vineyards, etofenprox and deltamethrin proved to be the best conventional insecticides, while pyrethrins were the most impactful among organic insecticides. Insecticide residual activity was evaluated in semi-field and field conditions. Acrinathrin showed the most significant residual effects in both conditions. In semi-field trials, most of the pyrethroids were associated with good results in terms of residual activity. However, these effects declined in field conditions, probably due to high temperatures. Organic insecticides showed poor results in terms of residual efficacy. Implications of these results in the context of Integrated Pest Management in conventional and organic viticulture are discussed.

Side Effects of Kaolin and Bunch-Zone Leaf Removal on Generalist Predators in Vineyards.

In vineyards, kaolin application and bunch-zone leaf removal (LR) were effective in the control of leafhoppers and Lobesia botrana, but their side effects on generalist predators are still poorly understood. In north-eastern Italian vineyards, the impact of kaolin and LR on species and functional diversity of spiders, as well as the abundance of spiders and generalist predatory insects, was assessed in one vineyard for two consecutive years and in two vineyards for one year. The ecological indices of the spider community were never influenced by kaolin and only in one case were they influenced by LR. At the spider family level, kaolin reduced the abundance of Araneidae, Oxypidae and Salticidae, but only in single cases. In single cases, kaolin reduced the amount of Orius sp. anthocorids and increased that of Scymninae coccinellids, whereas LR increased the amount of Aeolothrips sp. The moderate use of kaolin and the application of LR had negligible and inconsistent impacts on generalist predatory arthropods in vineyards and were therefore, compatible with IPM strategies.

Behavioural Evidence and Chemical Identification of a Female Sex Pheromone in Anagrus atomus (Hymenoptera: Mymaridae).

Anagrus atomus (L.) is an egg parasitoid involved in the biological control of Empoasca vitis (Göthe) in vineyards. Sex pheromones play a crucial role in mate finding for several parasitoid species and could be used for monitoring under field conditions. We carried out laboratory and field studies aimed at assessing the existence and identity of a possible A. atomus sex pheromone. We found that males were significantly attracted by virgin females independent of age. Males were not attracted to individuals of the same sex, but they were attracted by a crude extract from an unmated female and its polar fraction. Eugenol (4-allyl-2-methoxyphenol) was identified as the attractive substance and proved to be attractive not only in the olfactometer but also in another laboratory bioassay and under field conditions. Attraction of males, but not females, confirms that this is not an aggregation pheromone. This is the first sex-pheromone component identified in Mymaridae, however more compounds could be involved in the mating behaviour of A. atomus. The utility of a sex pheromone in A. atomus is discussed in the context of fitness returns.

Side Effects of Sulfur Dust on the European Grapevine Moth Lobesia botrana and the Predatory Mite Kampimodromus aberrans in Vineyards.

To reduce the impact of synthetic insecticides on human health and the environment, eco-friendly alternatives must be investigated. Knowledge of the side effects on pests and natural enemies of natural products applied to vineyards is very useful. Sulfur dust, which is used in vineyards to control powdery mildew, is investigated in laboratory and field bioassays for its effects on Lobesia botrana egg laying, egg hatching, and larval settlement. In field trials, the efficacy of sulfur dust against the two L. botrana carpophagous generations is compared with that of Bacillus thuringiensis and kaolin, and its side effects on the phytoseiid mite Kampimodromus aberrans are evaluated. In the bioassays, sulfur dust reduced female survival by 43%, egg laying by around 80%, egg hatching by 10%, and larval settlement by 55%. In field trials, sulfur dust caused a significant decrease in the number of L. botrana larval nests of both generations, even though the efficacy was lower than that of B. thuringiensis. No negative effects of sulfur dust on the predatory mite population density was observed. On the basis of these results, in the context of Integrated Pest Management strategies in vineyards, the activity of sulfur dust against L. botrana could be exploited by timing its application to the beginning of egg laying.

Relationships between Hyalesthes obsoletus, Its Herbaceous Hosts and Bois Noir Epidemiology in Northern Italian Vineyards.

Hyalesthes obsoletus is the vector of "Candidatus Phytoplasma (Ca. P.) solani," the causal agent of grapevine yellows Bois noir (BN). The relationships among the planthopper, its main herbaceous hosts as phytoplasma reservoirs (Convolvolus arvensis and Urtica dioica) and BN spreading were studied in northern Italy. In two areas the relationship between host plants and the phenology and survival of planthopper adults was investigated in potted plants and in field conditions. Moreover, H. obsoletus ecology, newly symptomatic grapevine occurrence and "Ca. P. solani" tuf-types' presence were studied in two vineyards (2014-2019). An earlier occurrence of H. obsoletus adults on C. arvensis than U. dioica and better adult survival of the originating host were observed. When U. dioica was prevalent, the vector occurred almost exclusively along the ditch outside the vineyard. Hyalesthes obsoletus amount varied widely from year to year and nymphal mortality due to late frosts was supposed. In one vineyard, the amount of newly symptomatic grapevines was significantly correlated with vector abundance in the previous year. The "Ca. P. solani" tuf-type was influenced by vector population levels on the two hosts. Since the abundance of H. obsoletus populations on the two hosts influences BN epidemiology and dynamics and the "Ca. P. solani" tuf-type, this must be considered in BN control strategies.

Is the existence of a mutualistic relationship between Lobesia botrana and Botrytis cinerea well-founded?

Some studies have highlighted benefits for Lobesia botrana by adding Botrytis cinerea mycelium to an artificial larval diet and have suggested a mutualistic relationship between the two organisms on grapevine, hypothesizing that fungal sterols were the nutritional factor involved. Because the nutritional quality of an artificial diet should be similar to grapes to allow extrapolation of the results to the field conditions, in the current study L. botrana larval performance was compared when larvae were fed on grapes (berries) or two artificial diets either with or without enrichment with B. cinerea. Based on sterol analysis, the two artificial diets had high cholesterol content, but relative to berries showed comparable and low phytosterol contents, respectively (high- and low-phytosterol, HPh, and LPh). While larval fitness on the HPh diet was similar to berries, the LPh diet led to higher mortality and worse larval performance. The addition of the fungus compensated for the shortage in the LPh diet but did not improve the HPh diet. Supplementing the LPh diet with linoleic acid, which is supplied also from B. cinerea, partially improved larval performance. In a field experiment, females did not show any egg-laying preferences towards naturally botrytized bunches. The positive effect of B. cinerea on the moth's next generation that is reported in the literature could be a consequence of fungus developed inside berry tunnels bored by larvae. Therefore, based on our data and previous reports the existence of a mutualistic relationship between L. botrana and B. cinerea is not well-founded.

Side Effects of Kaolin and Bunch-Zone Leaf Removal on Predatory Mite Populations (Acari: Phytoseiidae) Occurring in Vineyards.

The effects of kaolin and bunch-zone leaf removal on populations of predatory mites Kampimodromus aberrans (Oudemans) and Typhlodromus pyri Scheuten were assessed in the context of four trials (2015-2016) carried out against Lobesia botrana (Denis and Schiffermüller) (Lepidoptera: Tortricidae) in vineyards located in north-eastern Italy. Laboratory experiments were performed to evaluate the effects of kaolin on the survival and fecundity of K. aberrans and T. pyri populations originating from the same grape-growing area. In field trials, kaolin caused a gradual decrease in population density levels of both phytoseiid species (with the maximum reduction ranging from 49 to 91%) with a complete population recovery in the next spring. In laboratory experiments, kaolin was moderately harmful to both species, reducing their fecundity significantly (around 60%). Bunch-zone leaf removal determined lower phytoseiid populations in all trials, but this effect was significant only for K. aberrans in one of them. A limited use of kaolin and the adoption of bunch-zone leaf removal did not irreversibly affect phytoseiid populations in vineyards and thus can be considered compatible with IPM strategies.

Influence of Grapevine Cultivar on the Second Generations of Lobesia botrana and Eupoecilia ambiguella.

Grapevine cultivar can affect susceptibility to Lobesia botrana and Eupoecilia ambiguella with important implications on control strategies. A four-year study was carried out in north-eastern Italy on 10 cultivars (Cabernet Sauvignon, Carménère, Chardonnay, Merlot, Refosco dal Peduncolo Rosso, Rhine Riesling, Sauvignon Blanc, Terrano, Tocai Friulano and Verduzzo Friulano) grown in the same vineyard to assess whether the cultivar affects second-generation population levels of the two vine moths and L. botrana larval age composition. The influence of bunch traits measured at the peak of egg hatching on demographic parameters was also evaluated. Over the four years, L. botrana significantly prevailed over E. ambiguella in nine cultivars. Chardonnay and Tocai Friulano were the most infested cultivars and Merlot was the least infested. At the sampling date, the age composition of L. botrana varied with cultivar, with the larvae being significantly older on Chardonnay and younger on Carménère, Merlot and Verduzzo Friulano. Older larval age was significantly associated with higher bunch compactness. Larval infestation was not significantly influenced by either bunch compactness or berry volume, which suggested a more important role for contact and volatile substances mostly originating from the berries. These results allow for the improvement of Integrated Pest Management strategy against vine moths.

Control of Scaphoideus titanus with Natural Products in Organic Vineyards.

The leafhopper Scaphoideus titanus is the vector of 'Candidatus Phytoplasma vitis', the causal agent of Flavescence dorée (FD) a key disease for European viticulture. In organic vineyards, the control of S. titanus relies mostly on the use of pyrethrins that have suboptimal efficacy. During 2016, three field trials were conducted to evaluate the efficacy of kaolin, orange oil, insecticidal soap and spinosad against S. titanus nymphs, in comparison with pyrethrins. The activity of kaolin was evaluated also in the laboratory. In all field trials, kaolin had an efficacy against nymphs comparable to pyrethrins, while the other products were not effective. Laboratory results confirmed that kaolin increased nymph mortality. In organic vineyards, kaolin and pyrethrins are valuable tools in the management of FD. Nevertheless, their efficacy is lower compared to that of the synthetic insecticides used in conventional viticulture. Therefore, further research should be conducted in order to identify alternatives to synthetic insecticides for S. titanus control in the context of organic viticulture.

Efficacy and Mode of Action of Kaolin in the Control of Empoasca vitis and Zygina rhamni (Hemiptera: Cicadellidae) in Vineyards.

During 2015, the influence of kaolin applications and bunch-zone leaf removal on the grapevine leafhoppers, Empoasca vitis (Göthe) and Zygina rhamni Ferrari, and their egg parasitoids (Anagrus spp.) was tested in four vineyards of northeastern Italy. The mode of action of kaolin on E. vitis nymphs was also investigated in the laboratory. In the treated plots, kaolin was applied at a rate of 2% w/v on two occasions separated by 5-6 d. In two vineyards, it was applied either on the whole canopy or the bunch zone at the beginning of the E. vitis second generation (preventive criterion), and in the other two vineyards, it was applied to the whole canopy at the peak of the E. vitis third generation (curative criterion). Both the preventive and curative kaolin applications caused a significant decrease in the populations of E. vitis and Z. rhamni nymphs. The effect of the preventive applications was persistent and was associated with reduced E. vitis leaf symptoms. Kaolin did not influence the activity of Anagrus spp. Bunch-zone leaf removal did not affect leafhopper populations. Laboratory experiments showed that inhibition of feeding was the main mode of action through which kaolin affected nymph populations. Based on these outcomes, kaolin could be a valuable alternative to synthetic insecticides in controlling grapevine leafhoppers.

Influence of generation and photoperiod on larval development of Lobesia botrana (Lepidoptera: Tortricidae).

The influence of generation (under field conditions) and photoperiod (under laboratory conditions) on Lobesia botrana larvae development was studied. Some larvae were collected during three annual generations in two grape-growing areas of northeastern Italy, and others were individually reared in the laboratory from egg to pupa on an artificial diet under two different photoperiod conditions (respectively, daylight 16 h/d [long day {LD}] and 14 h/d [short day {SD}]). The mandible lengths of collected larvae were measured and the data analyzed morphometrically to determine the number of larval instars. In the laboratory study, the number of larval moultings, the mandible length of each instar, the development time from hatching larva to pupa, and the pupal weight were considered. The measurement of mandible lengths of larvae collected in the field indicated the existence of five larval instars in all three annual generations, but the size of the two oldest larval instars was significantly higher for third-generation larvae than for the previous generations. Under laboratory conditions, the larvae usually exhibited five instars, but the mandible lengths of larvae and the pupa size were greater for individuals reared under SD. These also took a greater number of days to develop from hatching larvae to pupae. Because a larger size of the final larval instar occurs in individuals that produce diapausing pupae under SD in both the laboratory and the field, a positive association between larval size and the probability of surviving the winter can be inferred.