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Molecular Identification of Cordyceps sinensis Related Fungi
Author(s) -
Zhu JiaShi,
Yao Yisang,
Guo Yinlan,
Liu Xinjie,
Wu Zimei
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a431-b
Subject(s) - cordyceps , biology , camellia sinensis , angelica sinensis , plant use of endophytic fungi in defense , genomic dna , dna , ribosomal dna , internal transcribed spacer , microbiology and biotechnology , botany , genetics , gene , ribosomal rna , phylogenetics , medicine , alternative medicine , pathology , traditional chinese medicine
Multiple fungi have been isolated and identified from natural Cordyceps sinensis, a precious tonic herb used in China for centuries. We reported simultaneous isolation at a temperature of 10–15 degree C of 2 major fungi, Hirsutella sinensis and Paecilomyces hepiali, from 200 pieces of fresh C. sinensis, supporting the multi‐fungi hypothesis for C. sinensis (FASEB J 19(5): A1033, 2005). We further confirmed P. hepiali and H. sinensis via molecular approaches in this study. Total rDNA was isolated from P. hepiali and H. sinensis. ITS segments of the DNA containing the 3¡ ¯ end of nuclear 18S rDNA, ITS1, 5.8S rDNA, and ITS2 and the 5¡ ¯ end of 28S rDNA were amplified by use of primers ITS4 (5‐TCCTCCGCTTATTGATATGC) and ITS5 (5‐GGAAGTAAAAGTCGTAACAAGG), followed by DNA sequencing. The ITS segments of the 2 fungi showed 73.6% sequence identity. The distinct regions of the sequences were identified and used to design “specific” primers for P. hepiali. The specificity of the “specific” primers was confirmed by amplifying DNA segments by PCR against P. hepiali DNA templates, but not against H. sinensis templates. Restriction enzymes were selected on the basis of ITS DNA sequences of P. hepiali and H. sinensis: Ava I and Alu I specific for H. sinensis, and Bam HI and Acc I specific for P. hepiali. The above results allow us to further examine the molecular existence of P. hepiali and H. sinensis in natural C. sinensis (on going).