Six components of partial resistance (RCs) were studied in 15 grapevine varieties with partial resistance to Plasmopara viticola: (i) infection frequency (ifR, proportion of inoculation sites showing sporulation), (ii) latent period (LP50, degree-days between inoculation and appearance of 50% of the final number of sporulating lesions), (iii) lesion size (LS, area of single lesions in mm 2), (iv) production of sporangia (SpoR, number of sporangia produced per lesion, and SpoR', number of sporangia produced per mm 2 of lesion), (v) infectious period (ip, number of sporulation events on a lesion), and (vi) infectivity of sporangia (INF, infection efficiency of sporangia produced on resistant varieties). Artificial inoculation monocycle experiments were conducted for a 3-year period on leaves collected at leaf development, flowering, and fruit development. Compared to the susceptible variety 'Merlot', the partially resistant varieties showed reduced ifR, longer Lp, smaller LS, fewer SpoR and SpoR', shorter ip, and lower INF. At leaf development, IFR, SPOR, and INF were higher and LP was shorter than at flowering and fruit development. RCs analysis through monocyclic experiments provides reliable assessments of the resistance response of grapevine accessions. The workload required for routine assessment in breeding programs could be reduced by measuring IFR and SPOR, while producing robust results. Downy mildew (DM) of grapevine is a serious disease caused by the obligate, biotrophic Oomycete Plasmopara viticola 1. P. viticola originated from North America, was introduced in Europe in the 1870 s 2 , and spread across the continent in the following years 1. The level of partial resistance to DM varies widely among Vitis species and cultivars 3,4. The Eurasian grapevine V. vinifera, which is widely cultivated for its agronomic and quality traits, is generally susceptible to DM, whereas Asian and American Vitis species (e.g., V. riparia and V. rupestris) show varying degrees of resistance 5 because of their coevolution with the pathogen. The partial resistance to P. viticola is conferred to grapevines by specific loci referred to as Rpvs (resistance to P. viticola) 6 ; partial resistance has also been found in V. vinifera germplasm 7. Breeding programs have been implemented based on the hybridization of American Vitis spp. with V. vinifera for the introgression of resistance genes into the domesticated background of V. vinifera. Some of the breeding lines, after several cycles of backcrossing, gained more than 80% of V. vinifera genetic background 8 , and incorporated one or more (pyramided) Rpvs. Some reports, which characterized partially resistant varieties under vineyard conditions, indicate a reduction in the speed of DM epidemics on resistant varieties in comparison with susceptible grapevine ones 9,10. Since the early 1900s, several partially resistant varieties with good grape quality and in some cases capable of producing a vinifera-like wine 11-13 have been selected and released by breeders. The resistance res...

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Although the use of biocontrol agents (BCAs) to manage plant pathogens has emerged as a sustainable means for disease control, global reliance on their use remains relatively insignificant and the factors influencing their efficacy remain unclear. In this work, we further developed an existing generic model for biocontrol of foliar diseases, and we parametrized the model for the Botrytis cinerea–grapevine pathosystem. The model was operated under three climate types to study the combined effects on BCA efficacy of four factors: (i) BCA mechanism of action, (ii) timing of BCA application with respect to timing of pathogen infection (preventative vs. curative), (iii) temperature and moisture requirements for BCA growth, and (iv) BCA survival capability. All four factors affected biocontrol efficacy, but factors iii and iv accounted for > 90% of the variation in model simulations. In other words, the most important factors affecting BCA efficacy were those related to environmental conditions. These findings indicate that the environmental responses of BCAs should be considered during their selection, BCA survival capability should be considered during both selection and formulation, and weather conditions and forecasts should be considered at the time of BCA application in the field.

Grapevine varieties showing partial resistance to downy mildew, caused by Plasmopara viticola, are a promising alternative to fungicides for disease control. Resistant varieties are obtained through breeding programs aimed at incorporating Rpv loci controlling the quantitative resistance into genotypes characterized by valuable agronomic and wine quality traits by mean of crossing. Traditional phenotyping methods used in these breeding programs are mostly based on the assessment of the resistance level after artificial inoculation of leaf discs in bioassays, by using the visual score proposed in the 2nd Edition of the International Organization of Vine and Wine (OIV) Descriptor List for Grape Varieties and Vitis species (2009). In this work, the OIV score was compared with an alternative approach, not used for the grapevine-downy mildew pathosystem so far, based on the measurement of components of resistance (RCs); 15 grapevine resistant varieties were used in comparison with the susceptible variety ‘Merlot’. OIV scores were significantly correlated with P. viticola infection frequency (IFR), the latent period for the downy mildew (DM) lesions to appear (LP50), and the number of sporangia produced per lesion (SPOR), so that when the OIV score increased (i.e., the resistance level increases), IFR and SPOR decreased, while LP50 increased. The relationship was linear for LP50, monomolecular for IFR and hyperbolic for SPOR. No significant correlation was found between OIV score and DM lesion size, sporangia produced per unit area of lesion, length of infectious period, and infection efficiency of the sporangia produced on DM lesions. The correlation between OIV score and area under the disease progress curve (AUDPC) calculated by using the RCs and a simulation model was significant and fit an inverse exponential function. Based on the results of this study, the measurement of the RCs to P. viticola in grapevine varieties by means of monocyclic, leaf disc bioassays, as well as their incorporation into a model able to simulate their effect on the polycyclic development of DM epidemics in vineyards, represents an improved method for phenotyping resistance level.

Grapevine downy mildew (DM) is caused by the dimorphic oomycete Plasmopara viticola , which incites epidemics through primary and secondary infection cycles that occur throughout the season. The secondary infection cycles are caused by the sporangia produced on DM lesions. The current research examined the relationship between numbers of airborne sporangia and DM development on grape leaves to determine whether spore sampler data can be useful to predict the potential for secondary infections of P. viticola . Three years (2015–2017) of spore sampler data confirmed that sporangia are a common component of the airborne microflora in a DM-infested vineyard and that their numbers depend on weather conditions. For a total of 108 days, leaf samples were collected from the vineyard at 2- to 3-day intervals and incubated under optimal conditions for P. viticola infection. The numbers of airborne sporangia sampled on 1 to 7 days before leaf sampling were significantly correlated with the numbers of DM lesions on the leaves. The best correlation (r=0.59), however, was found for the numbers of viable airborne sporangia (SPV), which were assessed by using equations driven by the vapour pressure deficit. In Bayesian and ROC curve analyses, SPV was found to be a good predictor of P. viticola infection of grape leaves, with AUROC=0.821 and false positive predictions mainly occurring at low SPV. A binary logistic regression showed that a threshold of 2.52 viable sporangia m -3 air day -1 enables a prediction of no infection with a posterior probability of 0.870, which was higher than the prior probability of 0.574. Numbers of viable sporangia in the vineyard air is therefore a useful predictor of infection and especially of no infection. The predictor missed some observed infections, but these infections were not severe (they accounted for only 31 of 374 DM lesions).

The parameterisation process of a previously developed modelling structure of the grapevine-downy mildew pathosystem is described. The model incorporates primary and secondary infections, host crop growth and development, along with a linkage between disease on foliage and disease on clusters. This process-based model was developed with a main objective of understanding the behaviour of the pathosystem under different, variable, environmental conditions, or under climate change. Six scenarios of disease conduciveness were developed in order to capture the range of environmental conditions under which potential downy mildew of grapevine epidemics can develop. These climate scenarios were based on moisture and temperature factors. The scenarios were translated into vectors of parameters for primary and secondary infections in the model. Model testing was performed in three steps: (i) an analysis of potential epidemics was conducted from the literature on grapevine downy mildew in order to delineate the behaviour of the pathosystem under different scenarios; (ii) a simulation experiment was conducted to investigate the response of the model to different patterns of environmental conditions, corresponding to six scenarios of disease conduciveness; and (iii) expected and simulated epidemics under these scenarios were compared. In scenarios, the model mobilised existing quantitative information on downy mildew of grapevine and generated outputs that are congruent with expected patterns of potential epidemic. This study indicates that the model is a reliable tool for simulating accurate and robust potential epidemics of downy mildew of grapevine in a scenario analysis. This can have many applications, such as the understanding of the behaviour of the pathosystem under climate change or when partial host resistance is involved.

A mechanistic model was developed to predict secondary infections of Plasmopara viticola and their severity as influenced by environmental conditions; the model incorporates the processes of sporangia production and survival on downy mildew (DM) lesions, dispersal and deposition, and infection. The model was evaluated against observed data (collected in a 3-year vineyard) for its accuracy to predict periods with no sporangia (i.e., for negative prognosis) or with peaks of sporangia, so that growers can identify periods with no/low risk or high risk. The model increased the probability to correctly predict no sporangia [P(P−O−) = 0.67] by two times compared to the prior probability, with fewer than 3% of the total sporangia found in the vineyard being sampled when not predicted by the model. The model also correctly predicted peaks of sporangia, with only 1 of 40 peaks unpredicted. When evaluated for the negative prognosis of infection periods, the model showed a posterior probability for infection not to occur when not predicted P(P−O−) = 0.87 with only 9 of 108 real infections not predicted; these unpredicted infections were mild, accounting for only 4.4% of the total DM lesions observed in the vineyard. In conclusion, the model was able to identify periods in which the DM risk was nil or very low. It may, therefore, help growers avoid fungicide sprays when not needed and lengthen the interval between two sprays, i.e., it will help growers move from calendar-based to risk-based fungicide schedules for the control of P. viticola in vineyards.

Grapevine downy mildew caused by Plasmopara viticola is one of the most important diseases in vineyards. Oospores that overwinter in the leaf litter above the soil are the sole source of inoculum for primary infections of P. viticola; in addition to triggering the first infections in the season, the oospores in leaf litter also contribute to disease development during the season. In the current study, a quantitative polymerase chain reaction (qPCR) method that was previously developed to detect P. viticola DNA in fresh grapevine leaves was assessed for its ability to quantify P. viticola oospores in diseased, senescent grapevine leaves. The qPCR assay was specific to P. viticola and sensitive to decreasing amounts of both genomic DNA and numbers of P. viticola oospores used to generate qPCR standard curves. When the qPCR assay was compared to microscope counts of oospores in leaves with different levels of P. viticola infestation, a strong linear relationship (R 2 = 0.70) was obtained between the numbers of P. viticola oospores per gram of leaves as determined by qPCR vs. microscopic observation. Unlike microscopic observation, the qPCR assay was able to detect significant differences between leaf samples with a low level of oospore infestation (25% infested leaves and 75% noninfested leaves) vs. samples without infestation, and this ability was not influenced by the weight of the leaf sample. The results indicate that the qPCR method is sensitive and provides reliable estimates of the number of P. viticola oospores in grapevine leaves. Additional research is needed to determine whether the qPCR method is useful for quantifying P. viticola oospores in grapevine leaf litter.