中国香菇自然群体遗传多样性的RAPD分析
摘要
用RAPD技术分析了采自我国13个省份、分属于8个不同植物区系区域的53个香菇菌株的遗传多样性。用10个随机引物共扩增出147条RAPD谱带。供试菌株在RAPD带型及DNA相似性上的差异表明，中国香菇自然群体具有相当丰富的遗传多样性，其中，横断山脉、云南高原、台湾及华南地区菌株的多样性尤为丰富。用平均连锁聚类法构建了遗传相关聚类图。以0.658的相似性为切割点，53个菌株可分为4大类群。类群Ⅰ和类群Ⅱ主要由横断山脉、云南高原和华中地区的菌株组成。类群Ⅲ包括了其余地区的菌株。类群Ⅳ则是由华北及四川省某些菌株组成的一个小分支。在0.750的相似性上，这4大类群可进一步分成20个RAPD指纹相似组(RGs)。其中，16个指纹相似组与菌株的地理来源呈明显相关，其余4个指纹相似组则属于广泛分布的类型。
Fifty-three strains of Lentinula edodes collected from 13 provinces distributed in 8 floristic regions of china were assessed for genetic diversity using the RAPD technique. A total of 147 RAPD bands were amplified using 10 random primers. Variations in RAPD banding patterns and DNA similarity among the strains revealed that there is significant genetic diversity in natural population of L.edodes in china, among which the diversity of those in the Hengduanshan, the Yunnan plateau, Taiwan and south china is more abundant. A clustering dendrogram was constructed using average linkage clustering. The 53 strains were classified into 4 groups using 0.658 similarity as a cut-off point. Group I and Group II mainly consisted of strains in the Hengduanshan, the Yunnan plateau and central china. GroupIII comprised those in the other regions. And groupIV was small branch composed of some strains in the north china and Sichuan Province. The 4 groups could subsequently be subdivided into 20 RAPD groups (RGs) at 0.750 similarity. Among them, sixteen of these RGs correlated strongly with the locality of strains, and the remaining 4 RGs belonged in widespread distributive types.
Fifty-three strains of Lentinula edodes collected from 13 provinces distributed in 8 floristic regions of china were assessed for genetic diversity using the RAPD technique. A total of 147 RAPD bands were amplified using 10 random primers. Variations in RAPD banding patterns and DNA similarity among the strains revealed that there is significant genetic diversity in natural population of L.edodes in china, among which the diversity of those in the Hengduanshan, the Yunnan plateau, Taiwan and south china is more abundant. A clustering dendrogram was constructed using average linkage clustering. The 53 strains were classified into 4 groups using 0.658 similarity as a cut-off point. Group I and Group II mainly consisted of strains in the Hengduanshan, the Yunnan plateau and central china. GroupIII comprised those in the other regions. And groupIV was small branch composed of some strains in the north china and Sichuan Province. The 4 groups could subsequently be subdivided into 20 RAPD groups (RGs) at 0.750 similarity. Among them, sixteen of these RGs correlated strongly with the locality of strains, and the remaining 4 RGs belonged in widespread distributive types.
引文
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Penn.state Univ
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[31]．陈春涛，浙江地区香菇栽培种DNA多态性研究，1995，河南农业大学硕士学位论文
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[34]. Kwan H.S.,Chiu S.W.,Pang K.M.&Cheng S.C.,Strain typing in Lentinula edodes by Polymerase Chain Reaction, Experimental Mycology,1992, 16:163～166
[35]. S.M.Udupa,F.Weigand,M.C.Saxena et al. Genotyping with RAPD and microsatellite makers resolves pathotype diversity in the ascochyta blight pathogen of chickpea,Theor Appl Genet 1998, 97:299～307
[36]. Welsh J, Peterson C,Mocelland M. Polymorphisms generated by arbitrarily primed PCR in the mouse:application to stain identification and genetic mapping Nucl Acids Res,1991,19:303～306
[37]. S.W.chiu, W.T.Chiu, F.C.Lin& D.Moore, Diversity of rDNA sequences indicates that China harbours the greatest germplasm resource of the cultivated mushroom Lentinula edodes, Mushroom Science, edited by
L.J.L.D,Van. Griensven, 2000, Ⅸ,Volume 1: 239～243
[38]. Lanner C, Bryngelsson T, Gustaffsson M. Genetic validity of RAPD makers at intrd-and inter-specific level in wild Brassica species with n=9.TAG,1996, 93:9～14
[39]. Nei, M et al, Mathematical mode for studying genetic uariation in terms of restriction endonucleases,1979,Proc. Natl. Acad. Sci.USA,76:5269～5273
[2]．张树庭，Miles P．6，食用蕈菌及其栽培，杨国良、张金霞等译，河北大学出版社，1992
[3]．杨新美等，食用菌栽培学，中国农业出版社1996：123～138
[4]．杨新美等，中国食用菌栽培学，农业出版社1988
[5]．杨新美等，中国食用菌栽培学，农业出版社1988
[6]．代江红，林芳灿等，香菇自然群体中个体间的空间分布和遗传相关，菌物系统，2001，(1)：100～106
[7]. Eric Kay and Rytas Vilgalys, Spital distribution and genetic relationships among individuals in a nature population of the oyster mushroom pleurotus ostreatus.Mycologia, 1992, 84 (2): 173～182
[8]．许东河、高忠、田清震等，中国一年生野生大豆群体的遗传多样性研究，应用与环境生物学报1999，5(5)：439～443
[9]．安贤惠、陈宝元、傅廷栋等，利用RAPD标记研究中国芥菜型油菜遗传多样性，华中农业大学学报1999，18(6)：524～526
[10]. ZHU You-Yong.WANG Yun-Yue等，The relationships betreen DNA Polymorphism and geographical origin of Australian Isolates of Verticillium Dahliae from cotton plants, 菌物系统.1999, 18 (3): 366～373
[11]．李泰辉、陈月琴、陆勇军等，鸡油菌的25SrDNA部分序列及其系统学意义，菌物系统，1999，18(1)：12～19
[12]. Karen W.Hughes.等, Pattern of geograpHic speciation in the genus Flammulina based on sequences of the ribosomal ITSI-5.8S-ITS2 area, Mycologia,1999.91(6): 978～986
[13]. Yilgalys, -R; sun, -B.L et al,Ancient and recent patterns of geograpHic
speciation in the Oyster mushroom Pleurotus revealed by pHylogenetic analysis of ribosomal DNA sequence,1994, 91(10): 4599～4603
[14]. Devid.hibbettd 等, Phylogenetic diversity in Shiitake inferred from nuclear ribosomal DNA sequences, Mycologia, 87(5):618～638.
[15]. Fukuda et al. Relationships of mitochondrial genome in TMI shiitake isolates based on FM analysis of distances,1994
[16]. Hibbett-DS. Et al. Phylogeny and biogeography of Lentinula inferred from an expanded rDNA dataset, Mycological-Research.1998,102(9):1041～1049
[17]. Fukuda. Et al. Phylogenetic analysis of Shiitake based on restriction fragment length polymorphisms of three regions of nuclear ribosomal DNA, 18S, 26S andinternal transcribed spacers: New information on Bornean wild strain. Reports of the Tottori Mycological Institute. December,1999,[print](37):36-49
[18]. Wiliams J.G.K, Kubelik A.R.and Lival K.J.,et al, DNA polymorphisms amplified by arbitrary primers are useful as genetic markers,.Nucleic Acidids Res,1990,18:6531～6535
[19]. Welsh J.,and McClell and M.O.,Fingerprinting genomes using PCR with arbitrary primers,Nucleic Acids Res,1990,18:7213～7218
[20]．曾伟，宋思扬，王泽生等，双孢蘑菇及大肥菇的种内及种间多态性RAPD分析，菌物系统，1999，18(1)：55～60
[21]. Chiu siu-wai Ai-Man Ma,Fang-can LIN, and David Moore,Genetic homogeneity of Cultivated Strains of Shiitake(Lentinula edodes) used in china,Mycological Research,1996,100 (11): 6393～6398
[22]．林芳灿，张树庭，中国栽培菌株不亲和性因子的分析，华中农业大学学报，1995，14(5)：459～466
[23]．汪中文，林芳灿，湖北省香菇自然种群交配型因子分析，食用菌学报，1996，3(4)：4～10
[24]. Mile P.G. and S.T.Chang,The collection and conservation of genes of Lentinus. Scentific and Technical Aspects of cultivating Edible fungi.The
Penn.state Univ
[25]．张树庭，林芳灿，蕈菌遗传与育种，中国农业出版社，1997
[26] 卯晓岚，中国菌物物种多样性研究与资源，1998，20：菌物专辑
[27]．钟月会，康亚男，练明总等，中国香菇交配型和基因型的分析，真菌学报，1992，11(4)：314～323
[28]．中国科学院《中国自然地理》编辑委员会，中国自然地理(植物地理)，科学出版社，1983
[29]. Zhang Y.F.,and Francis I.Molina,Studies on the Differentiation of Lentinula edodes Strains by RAPD Assay,食用菌学报，1994，1(1)：22～27
[30] 周文丽，中国香菇野生菌株及杂交组合DNA多态性研究，1996 河南农业大学硕士学位论文
[31]．陈春涛，浙江地区香菇栽培种DNA多态性研究，1995，河南农业大学硕士学位论文
[32]．萨姆布鲁克，E．F．弗里奇，T．曼尼阿蒂斯著，金冬雁，黎孟枫等译，侯云德等校，分子克隆实验指导，第二版，科学出版社，1993
[33] 余家林编，农业多元试验统计，北京农业大学出版社，1993
[34]. Kwan H.S.,Chiu S.W.,Pang K.M.&Cheng S.C.,Strain typing in Lentinula edodes by Polymerase Chain Reaction, Experimental Mycology,1992, 16:163～166
[35]. S.M.Udupa,F.Weigand,M.C.Saxena et al. Genotyping with RAPD and microsatellite makers resolves pathotype diversity in the ascochyta blight pathogen of chickpea,Theor Appl Genet 1998, 97:299～307
[36]. Welsh J, Peterson C,Mocelland M. Polymorphisms generated by arbitrarily primed PCR in the mouse:application to stain identification and genetic mapping Nucl Acids Res,1991,19:303～306
[37]. S.W.chiu, W.T.Chiu, F.C.Lin& D.Moore, Diversity of rDNA sequences indicates that China harbours the greatest germplasm resource of the cultivated mushroom Lentinula edodes, Mushroom Science, edited by
L.J.L.D,Van. Griensven, 2000, Ⅸ,Volume 1: 239～243
[38]. Lanner C, Bryngelsson T, Gustaffsson M. Genetic validity of RAPD makers at intrd-and inter-specific level in wild Brassica species with n=9.TAG,1996, 93:9～14
[39]. Nei, M et al, Mathematical mode for studying genetic uariation in terms of restriction endonucleases,1979,Proc. Natl. Acad. Sci.USA,76:5269～5273