Molecular Identification of Wild Cordyceps Asexual Isolates and Investigation of Their Genetic Diversity
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Abstract:
Cordyceps has high edible and medicinal values. Using nineteen wild Cordyceps isolates as the experimental material, internal transcribed spacer (ITS) universal primers were used for polymerase chain reaction (PCR) amplification and the products were sequenced. Cordyceps-related sequences were downloaded from GenBank to construct a phylogenetic tree, the enterobacterial repetitive intergenic consensus (ERIC)-PCR amplification of 19 Cordyceps asexual isolates was conducted, and the cordycepin content of fermentation broths of the isolates was determined. Phylogenetic analysis showed that all 19 Cordyceps asexual isolates were C. militaris. The average genetic distances of C. militaris from C. takaomontana, C. cicadae, C. gunnii, and C. sinensis were 0.1269, 0.1228, 0.2251, and 0.2354, respectively. In terms of genetic distance, C.militaris was relatively close to C. takaomontana, and far from C. sinensis. ERIC-PCR results showed that the 19 C. militaris strains could be divided into three distinct groups with a similarity coefficient of 0.8, and could also be divided into eight groups with a similarity coefficient of 0.9. Measurement of the cordycepin contents of the fermentation broths of the isolates showed that cordycepin could be detected in all broths, and the highest content reached 126.33 mg/L, which was 119 times as much as that of the existing production strain JD.
