Polymerase chain reaction-based assay for specific detection of Rhizoctonia solani AG-3 isolates.

摘要

R. solani AG-3 causes considerable yield losses in potato fields in eastern Canada and the USA. To obtain a fast and reliable identification method, RAPD experiments were carried out to obtain specific genetic markers of AG-3 isolates. DNA from various isolates of R. solani was submitted to RAPD amplification using random 10 mers primers. One of the 40 primers used yielded a 2.6 kbp fragment present in all isolates of AG-3. The specificity of this fragment was assessed by Southern blot analysis. Itwas partly sequenced and DNA primers were designed for PCR amplification experiments. A PCR-based restriction mapping method, using one restriction endonuclease, Xho I, was developed for specific detection of AG-3 isolates. Analysis of data showed thatAG-3 isolates were distinct to other AGs R. solani. This detection method is very specific and reliable and it can be applied on plant tissue and soil infected with R. solani AG-3.