A specific and sensitive PCR assay was developed for the detection and identification of Rhizoctonia solani AG-3, the main causal pathogen of stem canker and black scurf of potato. A conventional primer set (Rs1F2 and Rs2R1) was designed from the nuclear ribosomal internal transcribed spacer (ITS1 and ITS2) regions of R. solani . Following PCR amplification, a 0·5-kb product was amplified from DNA of all isolates of AG-3 using primers Rs1F2 and Rs2R1. No product was amplified when DNA from isolates belonging to a range of other R. solani anastomosis groups or from a selection of other potato pathogens was tested, confirming the specificity of the primers for AG-3 only. Rhizoctonia solani AG-3 was also detected in potato tissue with varying black scurf severity, and in soil inoculated with sclerotia of R. solani to a minimum detection level of 5 × 10 − 4 g sclerotia /g soil. In addition, specific primers RsTqF1 (based on the Rs1F2 sequence) and RsTqR1, and a TaqMan™ fluorogenic probe RQP1, were designed to perform real-time quantitative (TaqMan) PCR. The conventional PCR and real-time PCR assays were compared and combined with direct DNA extraction from soil and a seed-baiting method to determine the most reliable method for the detection and quantification of AG-3 in both artificially inoculated field soil and naturally infested soils. It was shown that direct DNA extractions from soil could be problematic, although AG-3 was detectable using this method combined with the real-time PCR assay. The amplification of Rhizoctonia solani by seed baiting increased the sensitivity of the assay compared with direct extraction of DNA from the soil, and AG-3 was detectable in artificially inoculated and naturally infested soils when seed baiting was combined with either the conventional PCR or the real-time PCR assay. The potential for using these rapid and quantitative AG-3-specific assays to address epidemiological questions and as tools for decision-making in disease management is discussed.

Chlorophyll a / b ratio did not respond to the applied treatment. Plants exposed to low UV-B transmission levels were visibly greener than those exposed to high transmission levels after approximately 23 d. The closest fits between the treatment and concentrations of UV-B screening pigments were associated with increased ambient biologically-weighted UV-B dose received by plants in the 5.5 h before each sampling, which coincided with the passage of the ozone hole over Rothera Point. As C. varians emerged from melting snow and ice, concentrations of the anthocyanin-like pigment and chlorophyll respectively increased and decreased faster in plants exposed to high transmission levels of UV-B, relative to those exposed to low UV-B transmission levels.2

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SummaryOptimum storage conditions to identify resistance to silver scurf among commercially grown potato cultivars were determined in a series of experiments. Inoculation of field-grown tubers with a conidial suspension of H. solani (concentration 104 conidia per ml of water) and incubation at 15 ~ with 95% RH for 1 month followed by 2 months at 85% RH produced the greatest differences in disease severity between potato cultivars. Results with glasshousegrown tubers were comparable with those from field-grown tubers, indicating that the test could be used to identify resistance in wild species of potato and to understand the inheritance of this resistance.