The high susceptibility of European grapevine cultivars to downy mildew (DM) and powdery mildew (PM) causes the intensive use of fungicides. Fungus-resistant cultivars (FRCs) with different resistance (R) loci have been bred and could play an important role in reducing plant protection treatments (PPTs). However, little information is available about the extent to which PPTs can be reduced in the field through the use of FRCs and the associated economic advantages. In this study, different strategies with reduced PPTs on FRCs were tested in field experiments. The results demonstrated that the number of PPTs can be reduced by 60 to 90%, resulting in reductions in applied copper and sulfur by 52 to 79% through the use of FRCs compared with susceptible cultivars, without affecting grape or plant health. The saving potential varied among years, depending on the type of R loci and climatic conditions. Furthermore, this study highlights that completely omitting PPTs in the cultivation of FRCs can result in PM or DM infections and possible loss of yield and fruit quality. In addition to the field experiments, a two-year observation of the performance of FRCs in commercial vineyards was undertaken, which highlighted not only the significant reduction in PPTs but also the financial savings that can be achieved through the use of FRCs.

Background The high susceptibility of European grapevine cultivars ( Vitis vinifera ) to downy mildew ( Plasmopara viticola) leads to the intensive use of fungicides in viticulture. To reduce this input, breeding programs have introgressed resistance loci from wild Vitis species into V. vinifera , resulting in new fungus-resistant grapevine cultivars (FRC). However, little is known about how these different resistance loci confer resistance and what the potential reduction in fungicide applications are likely to be if these FRCs are deployed. To ensure a durable and sustainable resistance management and breeding, detailed knowledge about the different defense mechanisms mediated by the respective Rpv ( Resistance to P. viticola ) resistance loci is essential. Results A comparison of the resistance mechanisms mediated by the Rpv3–1, Rpv10 and/or Rpv12 -loci revealed an early onset of programmed cell death (PCD) at 8 hours post infection (hpi) in Rpv12- cultivars and 12 hpi in Rpv10- cultivars, whereas cell death was delayed in Rpv3 -cultivars and was not observed until 28 hpi. These temporal differences correlated with an increase in the trans -resveratrol level and the formation of hydrogen peroxide shortly before onset of PCD. The differences in timing of onset of Rpv -loci specific defense reactions following downy mildew infection could be responsible for the observed differences in hyphal growth, sporulation and cultivar-specific susceptibility to this pathogen in the vineyard. Hereby, Rpv3- and Rpv12/Rpv3- cultivars showed a potential for a significant reduction of fungicide applications, depending on the annual P. viticola infection pressure and the Rpv -loci. Furthermore, we report on the discovery of a new P. viticola isolate that is able to overcome both Rpv3- and Rpv12 -mediated resistance. Conclusion This study reveals that differences in the timing of the defense reaction mediated by the Rpv3- , Rpv10 - and Rpv12 -loci, result in different degrees of natural resistance to downy mildew in field. Vineyard trials demonstrate that Rpv12/Rpv3- and Rpv3 -cultivars are a powerful tool to reduce the dependence of grape production on fungicide applications. Furthermore, this study indicates the importance of sustainable breeding and plant protection strategies based on resistant grapevine cultivars to reduce the risk of new P. viticola isolates that are able to overcome the respective resistance mechanism .

A wild grape haplotype (Rpv3-1) confers resistance to Plasmopara viticola. We mapped the causal factor for resistance to an interval containing a TIR-NB-LRR (TNL) gene pair that originated 1.6-2.6 million years ago by a tandem segmental duplication. Transient coexpression of the TNL pair in Vitis vinifera leaves activated pathogen-induced necrosis and reduced sporulation compared with control leaves. Even though transcripts of the TNL pair from the wild haplotype appear to be partially subject to nonsense-mediated mRNA decay, mature mRNA levels in a homozygous resistant genotype were individually higher than the mRNA trace levels observed for the orthologous single-copy TNL in sensitive genotypes. Allelic expression imbalance in a resistant heterozygote confirmed that cis-acting regulatory variation promotes expression in the wild haplotype. The movement of transposable elements had a major impact on the generation of haplotype diversity, altering the DNA context around similar TNL coding sequences and the GC-content in their proximal 5 '-intergenic regions. The wild and domesticated haplotypes also diverged in conserved single-copy intergenic DNA, but the highest divergence was observed in intraspecific and not in interspecific comparisons. In this case, introgression breeding did not transgress the genetic boundaries of the domesticated species, because haplotypes present in modern varieties sometimes predate speciation events between wild and cultivated species.

List of 2042 differentially expressed genes identified using interaction term analysis and displayed in Venn diagram (Fig. 7). xInteraction term analyses to identify DEG characteristic for a successful defence by comparing the pairwise contrasts of Rpv3–1 samples with susceptible samples. (adjusted P ≤ 0.1). FC, fold change. (XLSX 305 kb)

Background European grapevine cultivars (Vitis vinifera spp.) are highly susceptible to the downy mildew pathogen Plasmopara viticola. Breeding of resistant V. vinifera cultivars is a promising strategy to reduce the impact of disease management. Most cultivars that have been bred for resistance to downy mildew, rely on resistance mediated by the Rpv3 (Resistance toP.viticola) locus. However, despite the extensive use of this locus, little is known about the mechanism of Rpv3-mediated resistance. Results In this study, Rpv3-mediated defense responses were investigated in Rpv3(+) and Rpv3 grapevine cultivars following inoculation with two distinct P. viticola isolates avrRpv3(+) and avrRpv3, with the latter being able to overcome Rpv3 resistance. Based on comparative microscopic, metabolomic and transcriptomic analyses, our results show that the Rpv3-1-mediated resistance is associated with a defense mechanism that triggers synthesis of fungi-toxic stilbenes and programmed cell death (PCD), resulting in reduced but not suppressed pathogen growth and development. Functional annotation of the encoded protein sequence of genes significantly upregulated during the Rpv3-1-mediated defense response revealed putative roles in pathogen recognition, signal transduction and defense responses. Conclusion This study used histochemical, transcriptomic and metabolomic analyses of Rpv3(+) and susceptible cultivars inoculated with avirulent and virulent P. viticola isolates to investigate mechanism underlying the Rpv3-1-mediated resistance response. We demonstrated a strong correlation between the expressions of stilbene biosynthesis related genes, the accumulation of fungi-toxic stilbenes, pathogen growth inhibition and PCD.