Ana María Díez-Navajas

Grapevine downy mildew is a major disease in vineyards all around the world, caused by the oomycete Plasmopara viticola (Berk. & M. A. Curtis) Berl. & De Toni. Normally, its control depends almost exclusively on chemical and copper-based fungicides, especially in high-incidence areas with high relative humidity and mild temperatures. However, the European Union is determined to reduce the application of these phytochemicals by at least 50% by 2030, forcing winegrowers to seek alternative low-input strategies for proper sanitary maintenance. Basic substances (BSs), described in European Regulation (EC) 1107/2009, stand out as promising alternatives, but their molecular mechanism of action remains mostly unknown. In this context, this study analyzed the genetic effect in grapevine plants of several commercial products composed of BSs (chitosan, soy lecithin, Equisetum arvense and Salix cortex). All products exhibited promising results, triggering the induction of similar defence mechanisms, which included pathogenesis-related proteins (PRs), involved in direct pathogen repression; stilbenes, capable of producing antimicrobial compounds such as resveratrol and pterostilbene; several hormones, including oxylipins, ethylene, salicylic acid and terpenes, mediating immune signalling; and genes related to structural features of the plant, such as lignin, callose, cellulose and cuticular wax, constituting a first physiological barrier against P. viticola. Disease severity reduction differed among treatments, with Salix cortex showing the highest efficacy (58%), followed by BABA (38%) and LESOY (35%), while LECI and CHIT had minor effects (Salix cortex modulated the highest percentage of genes (41%), followed by natural infection without treatment (32%), LESOY (27%), BABA (26%), LECI (23%) and CHIT (23%). In terms of defence mechanisms, Salix cortex promoted the most pathways, LESOY induced eight, BABA and LECI seven and CHIT five. Overall, these results indicate that BSs can modulate several defence pathways in grapevine, supporting their potential use as sustainable alternatives for controlling downy mildew.

Plasmopara viticola is the causal agent of Grapevine Downy Mildew (GDM), which is a devastating disease of grapevines in humid temperate regions. The most employed method for protecting grapevines against GDM is the application of chemical fungicides. In Spain, Carboxylic Acid Amides (CAAs) are a fungicide group currently utilized in GDM control. In P. viticola, resistance to CAAs is conferred by G1105S and G1105V mutations in the CesA3 gene. Droplet digital polymerase chain reaction (ddPCR) is an innovative technique that combines PCR and droplet microfluidics to disperse the sample into thousands of water-in-oil droplets in which an amplification reaction is individually performed. In this study, we set up a ddPCR protocol to quantify S1105 and V1105 mutations conferring resistance to CAAs in P. viticola. The optimal PCR conditions were established, and the sensitivity and precision of the protocol were assessed. Four P. viticola populations coming from commercial vineyards in northern Spain were analyzed, and different allele frequencies were found in the analyzed samples corresponding to the different fungicide management strategies, ranging from 7.72% to 100%. Knowing the level of mutated alleles allows for designing resistance management strategies suited for each location. This suggests that similar ddPCR assays could be developed for studying mutations implicated in fungicide resistance in other fungicide groups and plant pathogens.

Grapevine, a crop of global economic importance, is annually affected by diseases that can compromise the quality and quantity of the harvest, producing large economic losses. Downy mildew caused by Plasmopara viticola (Berk. & M.A. Curtis) Berl. & de Toni is one of the most important diseases in the vineyard. To fight this pathogen, winegrowers often rely on conventional chemical fungicides or copper-based formulations, whose use is determined to be reduced by the European Commission due to their environmental consequences. Hence, alternative plant protection products (PPP) in grapevine must be considered and studied. In this context, we selected several alternative commercial products, based on basic substances (BS) or low-risk active substances (LRAS), to evaluate their suitability to deal with P. viticola. We measured the preventive activity of the products, both in vitro and in planta, as well as their toxicity against the sporangia and zoospores of the pathogen. Results showed that four commercial products were effective against the pathogen directly and preventively, being composed of approved basic substances, more concretely, chitosan, Equisetum arvense, lecithins, and Salix cortex. Among those, the products composed of lecithins and Salix cortex were the most toxic and active preventively. Therefore, these basic substances should be promoted in the vineyard as an alternative to conventional treatments in order to transition to a more sustainable viticulture.

The current study validated a mechanistic model for Botrytis cinerea on grapevine with data from 23 independent Botrytis bunch rot (BBR) epidemics (combinations of vineyards × year) that occurred between 1997 and 2018 in Italy, France, and Spain. The model was operated for each vineyard by using weather data and vine growth stages to anticipate, at any day of the vine-growing season, the disease severity (DS) at harvest (severe, DS ≥ 15%; intermediate, 5 < DS < 15%; and mild, DS ≤ 5%). To determine the ability of the model to account for latent infections, postharvest incubation assays were also conducted using mature berries without symptoms or signs of BBR. The model correctly classified the severity of 15 of 23 epidemics (65% of epidemics) when the classification was based on field assessments of BBR severity; when the model was operated to include BBR severity after incubation assays, its ability to correctly predict BBR severity increased from 65% to >87%. This result showed that the model correctly accounts for latent infections, which is important because latent infections can substantially increase DS. The model was sensitive and specific, with the false-positive and false-negative proportion of model predictions equal to 0.24 and 0, respectively. Therefore, the model may be considered a reliable tool for decision-making for BBR control in vineyards.

BACKGROUND: Control of Botrytis bunch rot (BBR) is currently based on the application of fungicides at four timings corresponding to specific growth stages of vines: end of flowering (A), pre‐bunch closure (B), veraison (C) and before harvest (D). The current research provides a network meta‐analysis of 116 studies conducted between 1963 and 2016 in nine countries, in which 14 strategies (based on combinations of 1, 2, 3, or 4 sprays applied in A, B, C, and/or D) were compared.RESULTS: When a one‐spray strategy was applied, BBR control was more effective with sprays applied in A, C, or D than B. With a two‐spray strategy, strategy AC provided similar control as strategy BC; strategy CD also provided good control. For a 3‐spray strategy, the best disease control was consistently obtained with strategy ACD. Four‐spray strategy ABCD provided the best control but often involved needless sprays so that the routine application of four sprays is not justified.CONCLUSIONS: Spraying at timing A seems to be very important for achieving efficient and flexible disease control. Flexibility is reduced by spraying at timing B rather than A. © 2018 Society of Chemical Industry

Lobesia botrana is the most significant pest of grape berries in Spain. Further knowledge of its phenology would enable wine growers to decide on an optimal treatment schedule. The aim of this study is, therefore, to predict the flight peaks of L. botrana in seven wine-growing regions of Spain. The main goal is to provide a prediction model based on meteorological data records. A logistic function model, based on temperature and humidity records, together with an exhaustive statistical analysis, were used to compare the wine-growing regions in which the male flight phenology of L. botrana displays similar patterns and to sort them into groups. By doing so, a joint study of the dynamics of the moth is possible in the regions within each group. A comparison of the prediction errors before and after applying the Touzeau model confirmed that the fit of the latter model is not sufficiently accurate for the regions under study. Moth flight predictions with the logistic function model are good, but accuracy may still be improved by evaluating other non-biotic and biotic factors.

Following inoculation of host and nonhost plants with Plasmopara viticola, the grapevine downy mildew, a histological survey was undertaken to identify the stage where its development is contained in nonhosts and in resistant host plants. Three herbaceous nonhost species, Beta vulgaris, Lactuca sativa, and Capsicum annuum, and three grapevine species displaying different level of resistance (Vitis vinifera [susceptible], Vitis riparia [partially resistant] and Muscadinia rotundifolia [totally resistant]) where inoculated by P. viticola using a controlled leaf disk inoculation bioassay. During the early steps of infection, defined as encystment of zoospores on stomata, penetration of the germ tube, and production of the vesicle with the primary hypha, there was no evidence of a clear-cut preference to grapevine tissues that could attest to host specificity. The main difference between host grapevine species and nonhosts was observed during the haustorium formation stage. In nonhost tissues, the infection was stopped by cell wall-associated defense responses before any mature haustorium could appear. Defense responses in resistant grapevines were triggered when haustoria were fully visible and corresponded to hypersensitive responses. These observations illustrate that, for P. viticola, haustorium formation is not only a key stage for the establishment of biotrophy but also for the host specificity and the recognition by grapevine resistance factors.