香菇菌株分子鉴别技术的分辨率比较 张瑞颖 黄晨阳 左雪梅 姜瑞波 王贺祥 张金霞 摘 要：本研究运用酯酶同工酶、RAPD、IGS1和IGS2 4种方法鉴别2个野生香菇菌株和19个栽培香菇菌株,并比较这4种鉴别方法的分辨率,结果表明:16条酯酶同工酶带中有15条具有多态性,将21个供试菌株分成11个类型,分辨率为0.92;10个随机扩增引物共扩增出86个DNA片段,其中95.3%具有多态性,将21个供试菌株分成16个类型,分辨率为0.97;IGS1将21个供试菌株分成7个类型,分辨率为0.81;IGS2将21个供试菌株分成7个类型,分辨率为0.73.这4种鉴别方法中RAPD的分辨率最高,与这4种鉴别方法的综合分析结果相同,因此RAPD可以作为香菇菌株鉴别的可靠依据. 关键词：酯酶同工酶;随机扩增多态性;转录单位间隔区 分类号：Q939.96 文献标识码：A 文章编号：1672-6472(2005)04-0517-0524 DISCRIMINATORY POWER OF MOLECULAR TYPING TECHNIQUES FOR LENTINULA EDODES ZHANG Rui-Ying HUANG Chen-Yang ZUO Xue-Mei JIANG Rui-Bo WANG He-Xiang ZHANG Jin-Xia 基金项目：北京市科委项目(D0705007040291)和国家科技基础条件平台工作项目(2004DKA30560) 作者单位：张瑞颖（中国农业科学院农业资源与农业区划研究所,北京,100081） 黄晨阳（中国农业科学院农业资源与农业区划研究所,北京,100081） 左雪梅（中国农业科学院农业资源与农业区划研究所,北京,100081） 姜瑞波（中国农业科学院农业资源与农业区划研究所,北京,100081） 王贺祥（中国农业大学生物学院,北京,100094） 张金霞（中国农业科学院农业资源与农业区划研究所,北京,100081） 参考文献： [1]Albee SR, Mueller GM, Kropp BR, 1996. Polymorphisms in the large intergenic spacer of the nuclear ribosomal repeat identify Laccaria proxima strains. Mycologia, 88: 970～976 [2]Bowden CG, Toyse DJ, 1991. Linkage relationships of allozyme-encoding loci in shiitake, Lentinula edodes. Genome, 34:652～657 [3]Chiu SW, Ma AM, Lin FC, Moore D, 1996. Genetic homogeneity of cultivated strains of shiitake (Lentinula edodes) used in China as revealed by the polymerase chain reaction. Mycological Research, 100: 1393～1399 [4]Chiu SW, Wang ZM, Chiu WT, Lin FC, Moore D, 1999. An integrated study of individualism in Lentinula edodes in nature and its implication for cultivation strategy. Mycological Research, 103:651～660 [5]He DM, Chen MJ, Tan Q, 1998. Analysis of esterase isozyme in mycelium of Lentinula edodes growing in different media. Acta Edulis Fungi, 5:18～20 (in Chinese) [6]Huang ZL, Xie BG, Xie FQ, Fu RZ, 2002. Molecular identification of 15 Lentinula edodes strains cultivated with artificial synthetic compost. Acta Edulis Fungi, 9:5～8 (in Chinese) [7]Hunter PR, 1990. Reproducibility and indices of discriminatory power of microbial typing methods. Journal of Clinical Microbiology, 28:1903～1905 [8]Kazuhisa T, Teruyuki M, 2004. Strain typing of shiitake (Lentinula edodes) cultivars by AFLP analysis, focusing on a heat-dried fruiting body. Mycoscience, 45:79～82 [9]Ma FY, Luo XC, 2002. Application of molecular markers in genetic and breeding of edible mushroom. Mycosystema, 21:147～151 (in Chinese ) [10]Micaies JA, Bonde MR, Peterson GL, 1986. The use of isozyme analysis in fungal taxonomy and genetics. Mycotaxon, 12:405～449 [11]Rajiv K, 1991. DNA polymorphisms in Lentinula edodes, the shiitake mushroom. Applied and Environmental Microbiology, 57:1735～1739 [12]Royse DJ, May B, 1987. Identification of shiitake genotypes by multilocus enzyme electrophoresis: catalog of lines. Biochemical Genetics, 25:705～717 [13]Saito T, Tanaka N, Shinozawa T, 2002. Characterization of subrepeat regions within rDNA intergenic spacers of the edible basidiomycete Lentinula edodes. B ioscience, Biotechnology, and Biochemistry, 66:2125～2133 [14]Toyomasu T, Zennyozi A, 1981. On applicaton of isoenzyme electrophoresis to identification of strains in Lentinus edodes (shiitake).Mushroom Science, 11:675～684 [15]Wang JZ, 2002. Protein Methods. Beijing: Science Press (in Chinese). 150～200 [16]Wang ZR, 1996. Plant Allozyme Analysis. Beijing: Science Press (in Chinese). 90～134 [17]Zhang Y, Molina F, 1995. Strain typing of Lentinula edodes by random amplified polymorphic DNA assay. FEMS Microbiology Letters,131:17～20 [18]贺冬梅,陈明杰,谭琦,1998.不同培养基对香菇菌丝体酯酶同工酶的影响.食用菌学报,5:18～20 [19]黄志龙,谢宝贵,谢福泉,傅瑞洲,2002.15个代料栽培香菇菌株的分子鉴别.食用菌学报,9:5～8 [20]马富英,罗信昌,2002.分子标记在食用蕈菌遗传育种中的应用.菌物系统,21:147～151 [21]汪家政,范明,2002.蛋白质技术手册.北京:科学出版社.150～200 [22]王中仁,1996.植物等位酶分析.北京:科学出版社.90～134 菌物学报 2005年第4期-HTM文件 No.8
