The genomics era brought unprecedented opportunities for genetic analysis of host resistance, but it came with the challenge that accurate and reproducible phenotypes are needed so that genomic results appropriately reflect biology. Phenotyping host resistance by natural infection in the field can produce variable results due to the uncontrolled environment, uneven distribution and genetics of the pathogen, and developmentally regulated resistance among other factors. To address these challenges, we developed highly controlled, standardized methodologies for phenotyping powdery mildew resistance in the context of a phenotyping center, receiving samples of up to 140 grapevine progeny per F family. We applied these methodologies to F families segregating for REN1- or REN2-mediated resistance and validated that some but not all bioassays identified the REN1 or REN2 locus. A point-intercept method (hyphal transects) to quantify colony density objectively at 8 or 9 days postinoculation proved to be the phenotypic response most reproducibly predicted by these resistance loci. Quantitative trait locus (QTL) mapping with genotyping-by-sequencing maps defined the REN1 and REN2 loci at relatively high resolution. In the reference PN40024 genome under each QTL, nucleotide-binding site-leucine-rich repeat candidate resistance genes were identified-one gene for REN1 and two genes for REN2. The methods described here for centralized resistance phenotyping and high-resolution genetic mapping can inform strategies for breeding resistance to powdery mildews and other pathogens on diverse, highly heterozygous hosts.
Growth room experiments were conducted to assess the interaction of soil type, biofungicides, soil compaction and pathotype/host on infection and symptom development caused by Plasmodiophora brassicae, the cause of clubroot on Brassica spp. In two initial experiments, four soil types (peat soil, mineral soil, non‐calcareous sand, soil‐less mix), two biofungicides (Bacillus subtilis, Clonostachys rosea), and two pathotypes (3 and 6, Williams’ differential set) were assessed. Differences in clubroot severity associated with soil type were unexpectedly small and variable. Prestop (C. rosea) was often more effective than Serenade (B. subtilis) at reducing clubroot levels on peat and mineral soils, but less effective than Serenade on sand. Inoculation with pathotype 3 often resulted in a slightly higher mean severity than pathotype 6. The interaction of soil type × biofungicide was similar on both canola (B. napus) and Shanghai pak choy (B. rapa subsp. chinensis), whether the soil was kept saturated or allowed to drain after inoculation. The impact of soil type on biofungicide efficacy might explain, in part, why biofungicides are more effective in one location than another. The observation that clubroot severity in soil‐less mix was affected by compaction led to an investigation of soil bulk density. Severity was higher in soil‐less mix that was more compacted than in the initial experiments, and was lower in peat and mineral soils when soil bulk density was reduced by adding soil‐less mix. In this study, soil bulk density had a larger impact on clubroot than soil type, organic matter or pathotype.
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