野生虫草无性型分离株分子鉴定及遗传多样性的研究  

Molecular Identification of Wild Cordyceps Asexual Isolates and Investigation of Their Genetic Diversity

作　　者：冯德龙[1,2] 胡惠萍[2] 黄龙花[2] 谢意珍[2] 吴清平[2] 霍光华[1] 

机构地区：[1]江西农业大学生物科学与工程学院,江西南昌330045 [2]广东省微生物研究所省部共建华南应用微生物国家重点实验室广东省菌种保藏与应用重点实验室广东省微生物新技术公共实验室,广东广州510070

出　　处：《现代食品科技》2017年第4期108-113,167,共7页Modern Food Science and Technology

基　　金：广东省科技计划农业攻关项目(2012A020100010);国家科技支撑计划项目(2013BAD16B05);国家自然基金项目(31560574)

摘　　要：虫草具有较高的食用与药用价值,本文以19株野生虫草分离株为研究材料,利用ITS引物进行PCR扩增、测序,并从Genbank上下载虫草属相关序列,构建系统发育树,并对19株虫草无性型分离物进行ERIC-PCR扩增,同时还检测了上述无性分离物发酵液中虫草素的含量。系统发育分析表明在分子水平上19株虫草的无性型分离菌株,都为蛹虫草(Cordyceps militaris)。与高雄山虫草(C.takaomontana)、蝉花(C.cicadae)、古尼虫草(C.gunnii)以及冬虫夏草(C.sinensis)平均遗传距离分别为0.1269、0.1228、0.2251和0.2354,在遗传距离上,与高雄山虫草和蝉花较近,与冬虫夏草较远。ERIC-PCR相似系数在0.8水平上,可以将19个蛹虫草菌株分为3组,相似系数在0.9水平上19株蛹虫草菌种可以分为8组。对蛹虫草发酵液中虫草素含量检测发现,所有蛹虫草无性型分离物发酵液都含有虫草素,最高的可达到126.33 mg/L,为现有的蛹虫草生产菌株JD的119倍。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 Gen Bank 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. militarisstrains 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.

关 键 词：虫草 鉴定 遗传多样性 虫草素 

分 类 号：S567.3[农业科学—中草药栽培]