Study in molecular systematics of Agaricus blazei Murill

• Main Authors: Huang, Ya-Hui, 黃雅惠
• Other Authors: 許瑞祥
• Format: Others
• Language: zh-TW
• Published: 1999
• Online Access: http://ndltd.ncl.edu.tw/handle/02959179255164814907

碩士 === 國立臺灣大學 === 農業化學研究所 === 87 === Agaricus blazei Murill a mass production target for the biotechnology industry, is developing edible mushroom with the function of nutrition, favor and physiological regulation. The polysaccharides from A. blazei was known for its highest anti-tumor activity in mushroom. After long-term artificial cultivation, A. blazei has developed obviously distinct morphology and chemical composition in different locations. To realize of the relationships of A. blazei with related species, this thesis focused on the classification and identification of A. blazei using molecular techniques. The genus of Agaricus was distinguished from others by the parsimony analysis from nucleotide sequence of the small subunit ribosomal RNA gene (NS3/NS4). Based on phylogenetic analysis of the divergent domain D2 of the large subunit ribosomal RNA gene (LSU-D2) and of the internal transcribed spacers (ITS), A. blazei and A. sylvaticus were considered as the same species. For rapid identification of A. blazei, a species-specific polymerase chain reaction (PCR) technique was developed using a universal primer, NS7 and a primer Bla designed from the specific sequence of ITS1 in A. blazei. The minimal concentration of DNA required for PCR was 10 pg/μl of DNA for PCR. A. blazei and A. sylvaticus could be clearly differentiate by genetic fingerprint using random amplified polymorphic DNA (RAPD) produced from the random primers, OPB15, OPB18 and R4.