Molecular marker assisted detection of the mycoparasite <i>Coniothyrium minitans</i> A69 in soil

Abstract

Coniothyrium minitans A69 has been shown to have biological control activity against the plant pathogen Sclerotinia sclerotiorum and a PCR based assay has been developed to specifically identify this isolate The practical application of this PCR assay for detection of C minitans from soil was assessed Sterile and nonsterile soil was inoculated with spores from C minitans A69 at five different concentrations and DNA recovered using a SDS/Phenol/Chloroform method A number of factors affected DNA recovery and subsequent PCR with a maximum sensitivity of down to 1x10² spores/g soil achieved in sterile soil Detection of C minitans in nonsterile soil was hampered by failure of the fungus to germinate However this method has improved throughput and cost effectiveness compared with conventional detection methods involving quantitative colony recovery