香菇菌株分子鑒別技術的分辨率比較 張瑞穎　 黃晨陽　 左雪梅　 姜瑞波　 王賀祥　 張金霞　 摘　要：本研究運用酯酶同工酶、RAPD、IGS1和IGS2 4種方法鑒別2個野生香菇菌株和19個栽培香菇菌株,并比較這4種鑒別方法的分辨率,結果表明:16條酯酶同工酶帶中有15條具有多態性,將21個供試菌株分成11個類型,分辨率為0.92;10個隨機擴增引物共擴增出86個DNA片段,其中95.3%具有多態性,將21個供試菌株分成16個類型,分辨率為0.97;IGS1將21個供試菌株分成7個類型,分辨率為0.81;IGS2將21個供試菌株分成7個類型,分辨率為0.73.這4種鑒別方法中RAPD的分辨率最高,與這4種鑒別方法的綜合分析結果相同,因此RAPD可以作為香菇菌株鑒別的可靠依據. 關鍵詞：酯酶同工酶;隨機擴增多態性;轉錄單位間隔區 分類號：Q939.96　文獻標識碼：A 文章編號：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　 基金項目：北京市科委項目(D0705007040291)和國家科技基礎條件平臺工作項目(2004DKA30560) 作者單位：張瑞穎（中國農業科學院農業資源與農業區劃研究所,北京,100081）　 　　　　　黃晨陽（中國農業科學院農業資源與農業區劃研究所,北京,100081）　 　　　　　左雪梅（中國農業科學院農業資源與農業區劃研究所,北京,100081）　 　　　　　姜瑞波（中國農業科學院農業資源與農業區劃研究所,北京,100081）　 　　　　　王賀祥（中國農業大學生物學院,北京,100094）　 　　　　　張金霞（中國農業科學院農業資源與農業區劃研究所,北京,100081）　 參考文獻： [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]賀冬梅,陳明杰,譚琦,1998.不同培養基對香菇菌絲體酯酶同工酶的影響.食用菌學報,5:18～20 [19]黃志龍,謝寶貴,謝福泉,傅瑞洲,2002.15個代料栽培香菇菌株的分子鑒別.食用菌學報,9:5～8 [20]馬富英,羅信昌,2002.分子標記在食用蕈菌遺傳育種中的應用.菌物系統,21:147～151 [21]汪家政,范明,2002.蛋白質技術手冊.北京:科學出版社.150～200 [22]王中仁,1996.植物等位酶分析.北京:科學出版社.90～134 菌物學報 2005年第4期-HTM文件 No.8