Development of the specific rDNA-ITS and SSR primers to detect the ectomycorrhizal fungus Rhizopogon roseolus

Abstract

　By the comparison of sequences of rDNA-ITS (internal transcribed spacer) region and SSR (simple sequence repeat) loci from 45 isolates of Rhizopogon roseolus, the five specific primer sets, one rDNA-ITS primer and four SSR primers, were designed for detection of R. roseolus. All these primer sets amplified the specific PCR products for the total DNAs from mycelia of 45 R. roseolus isolates, respectively, but no amplified product was detected for the DNAs from host plant (Pinus thunbergii) and 16 ectomycorrhizal isolates consist of 13 species including two Rhizopogon species, R. luteolus and R. superiorensis. Furthermore, target regions of these primer sets were also specifically amplified using by DNA templates from ectomycorrhizas of this fungus. Thus, the five sets of primer pair represent a potent tool for the monitoring of introduced isolates of R. roseolus and for molecular ecology applications.