Analysis of Diversity in Pythium aphanidermatum Populations from a Single Greenhouse Reveals Phenotypic and Genotypic Changes over 2006 to 2011

A study was conducted to investigate phenotypic and genotypic changes within Pythium aphanidermatum populations during the period 2006 to 2011. In total, 92 isolates of P. aphanidermatum (59 in 2006 and 33 in 2011) were obtained from different planting sites (soil) of cucumber from a single greenhouse. Generated sequences of the internal transcribed spacer (ITS) ribosomal DNA showed that all, except one isolate, share an identical sequence of the ITS region. Most (89%) P. aphanidermatum isolates were found to be aggressive on cucumber seedlings, with no significant differences in the aggressiveness level between populations obtained from different planting rows or different years. Sensitivity to metalaxyl among populations of P. aphanidermatum increased significantly from concentration resulting in 50% growth inhibition levels of 0.070 to 1.823 (average 0.824 μg ml ) in 2006 to 0.002 to 0.564 (average 0.160 μg ml ) in 2011. Amplified fragment length polymorphism analysis of the 92 isolates produced 92 different genotypes and 985 polymorphic loci. P. aphanidermatum populations from 2006 and 2011 were found to have low levels of genetic diversity (H = 0.1425), which implies introduction of the isolates into the greenhouse via common sources. Results from analysis of molecular variance (F = 0.0307 in 2006 and 0.0222 in 2011) provided evidence for frequent exchange of Pythium inoculum between different planting locations within the same year. However, the analysis showed moderate levels (F = 0.1731) of genetic differentiation among populations from the 2 years. This was supported by unweighted pair group method with arithmetic means analysis, which showed clustering of many of the 2006 isolates in separate clusters. The change in the metalaxyl sensitivity of the populations from 2006 to 2011 accompanied by the genetic differences among these two populations may suggest that many of the isolates from 2006 were lost and were replaced by new and highly sensitive P. aphanidermatum isolates by 2011.