26个草菇菌株SRAP分析
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摘要
为充分利用和保护草菇种质资源,笔者利用SRAP-PCR技术对来源于不同地区的26个草菇菌株进行遗传多样性分析,采用改进的SDS法提取DNA。由9条正向引物和17条反向引物组成153对引物中,筛选出11对多态性引物,共扩增出148个位点,其中多态性位点142个,片段大小在0.11～2.51 kb之间,扩增位点多态性比率在75.0%～100%之间,多态性比率为95.5%。利用NTSYS-PC2.1e软件对26个草菇品种进行聚类分析,标记在相似系数0.76时,26个草菇菌株可划分为3个类群。其中,天达V971、高邮草V112、单生草菇等19个菌株聚为一类;聚为第二类的6个菌株包含绵草V23、习酒草V11、绵草931、习酒V23、高邮V11、土草V23;而银丝草菇自成一类。说明草菇存在相互引种后又重新保藏命名的现象,同时草菇在一定程度上出现了地域基因分化。探究草菇分子基因库,为山东省草菇种质资源库的建立、草菇育种和遗传改良提供了基础数据。
To fully utilize and protect the germplasm resources of Volvariella volvacea, the authors analyzed the genetic diversity of 26 V. volvacea strains from different provinces and regions by using SRAP-PCR, andextracted DNA adopting the improved SDS method. Among 153 pairs of primers composed of 9 positiveprimers and 17 reverse primers, 11 pairs of polymorphic primers were screened and 148 polymorphic loci wereamplified, of which 142 were polymorphic loci, the fragment size was between 0.11-2.51 kb, the ratio ofpolymorphic loci was between 75.0%-100%, and the polymorphism ratio was 95.5%. Cluster analysis of 26 V.volvacea varieties was carried out by NTSYS-PC2.1 e software, the results showed that: when the similaritycoefficient was 0.76, 26 strains of V. volvacea could be divided into 3 groups; among them, 19 strains of TiandaV971, Gaoyou grass V112 and single V. volvacea were clustered into one class, and 6 strains of the secondgroup included V23, V11, 931, V23, Gaoyou V11, and V23, and silver fillet V. volvacea was clustered into a class. It indicates that after being introduced to each other, V. volvacea is re-preserved and named. At the sametime, V. volvacea has regional gene differentiation to some extent. Exploring the molecular gene pool of V.volvacea could provide basic data for the establishment of V. volvacea germplasm resource bank, and the breeding and genetic improvement of V. volvacea in Shandong.
To fully utilize and protect the germplasm resources of Volvariella volvacea, the authors analyzed the genetic diversity of 26 V. volvacea strains from different provinces and regions by using SRAP-PCR, andextracted DNA adopting the improved SDS method. Among 153 pairs of primers composed of 9 positiveprimers and 17 reverse primers, 11 pairs of polymorphic primers were screened and 148 polymorphic loci wereamplified, of which 142 were polymorphic loci, the fragment size was between 0.11-2.51 kb, the ratio ofpolymorphic loci was between 75.0%-100%, and the polymorphism ratio was 95.5%. Cluster analysis of 26 V.volvacea varieties was carried out by NTSYS-PC2.1 e software, the results showed that: when the similaritycoefficient was 0.76, 26 strains of V. volvacea could be divided into 3 groups; among them, 19 strains of TiandaV971, Gaoyou grass V112 and single V. volvacea were clustered into one class, and 6 strains of the secondgroup included V23, V11, 931, V23, Gaoyou V11, and V23, and silver fillet V. volvacea was clustered into a class. It indicates that after being introduced to each other, V. volvacea is re-preserved and named. At the sametime, V. volvacea has regional gene differentiation to some extent. Exploring the molecular gene pool of V.volvacea could provide basic data for the establishment of V. volvacea germplasm resource bank, and the breeding and genetic improvement of V. volvacea in Shandong.
引文
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[9]宫志远,任海霞,姚强,等.35个山东主栽平菇菌株的ISSR遗传差异分析[J].基因组学应用生物学,2010,29(3):507-512.
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[12]任海霞,宫志远,蒋志琴,等.26个香菇菌株的遗传多样性分析[J].山东农业科学,2017,49(11):20-23.
[13]秦莲花,宋春艳,谭琦,等.用ITS和ISSR分子标记技术鉴别香菇生产用种[J].菌物学报,2006,25(1):94-100.
[14]彭方仁,吴莺莺,郝明灼,等.利用ISSR和SRAP标记分析油茶遗传多样性[J].南京林业大学学报:自然科学版,2012,36(5):19-25.
[15]唐源江,曹雯静,吴坤林.基于SRAP标记的国兰种质资源遗传多样性分析及分子身份证构建[J].中国农业科学,2015,48(9):1795-1806.
[16]李亚利,于铁峰,罗爱玉,等.SRAP分子标记在茄果类蔬菜航天诱变育种中的应用[J].中国农学通报,2015,31(28):60-64.
[17]蔡丹凤,陈美元,蔡志欣,等.鸡腿蘑栽培菌株的SRAP分析[J].食用菌学报,2010,17(3):12-15.
[18]徐凯,唐传红,王天娇,等.基于SRAP,ISSR和RAPD分析灵芝G0130菌株单核体多态性[J].工业微生物,2014,44(4):39-45.
[10]温志强,熊芳,陈吉娜,等.分子标记鉴别灰树花种质资源的研究[J].热带作物学报,2011,32(7):1330-1336.
[20]江玉姬,邓优锦,刘新锐,等.74株草菇遗传多样性分析[J].核农学报2015,29(11):2084-2092.
[21]韦仕岩,吴圣进,汪茜,等.草菇菌株的ISSR遗传差异分析[J].热带作物学报,2013,34(11):2209-2213.
[2]张树庭,游中骥.草菇及其栽培[M].香港:香港艺美图书公司出版,1978.
[3]胡显海,周晓飞.草菇高产栽培技术[J].现代农业科技,2006(21):55-58.
[4]李丹青,王杰.草菇菌种退化相关分子标记的筛选[J].西北农林科技大学学报:自然科学版,2015,43(8):195-201.
[5]傅俊生,刘新锐,谢宝贵,等.草菇SCAR遗传标记建立及其杂种鉴定应用[J].中国农学通报,2010,26(17):41-46.
[6] Li G, Quiros C F. Sequence-related amplified polymorphism(SRAP), a new marker system based on a simple PCR reaction:its application to mapping and gene tagging in Brassica[J].Theoretical and Applied Genetics,2001,103(2/3):455-461.
[7]蔡志欣,蔡丹凤,陈美元,等.32个茯苓菌株的SRAP分析[J].食药用菌,2013(2):96-98.
[8]王景雪,孙毅,高武军,等.一种简便实用的植物总DNA提取方法[J].山西大学学报:自然科学版,2000,23(3):271-272.
[9]宫志远,任海霞,姚强,等.35个山东主栽平菇菌株的ISSR遗传差异分析[J].基因组学应用生物学,2010,29(3):507-512.
[10]郭仲杰.双孢蘑菇栽培菌株的农艺学性状与DNA指纹分析[D].福州:福建农林大学,2009.
[11]应正河,林衍铨,马璐,等.绣球菌栽培菌株RAPD和SRAP遗传多样性分析[J].西南农业学报,2014,27(2):777-780.
[12]任海霞,宫志远,蒋志琴,等.26个香菇菌株的遗传多样性分析[J].山东农业科学,2017,49(11):20-23.
[13]秦莲花,宋春艳,谭琦,等.用ITS和ISSR分子标记技术鉴别香菇生产用种[J].菌物学报,2006,25(1):94-100.
[14]彭方仁,吴莺莺,郝明灼,等.利用ISSR和SRAP标记分析油茶遗传多样性[J].南京林业大学学报:自然科学版,2012,36(5):19-25.
[15]唐源江,曹雯静,吴坤林.基于SRAP标记的国兰种质资源遗传多样性分析及分子身份证构建[J].中国农业科学,2015,48(9):1795-1806.
[16]李亚利,于铁峰,罗爱玉,等.SRAP分子标记在茄果类蔬菜航天诱变育种中的应用[J].中国农学通报,2015,31(28):60-64.
[17]蔡丹凤,陈美元,蔡志欣,等.鸡腿蘑栽培菌株的SRAP分析[J].食用菌学报,2010,17(3):12-15.
[18]徐凯,唐传红,王天娇,等.基于SRAP,ISSR和RAPD分析灵芝G0130菌株单核体多态性[J].工业微生物,2014,44(4):39-45.
[10]温志强,熊芳,陈吉娜,等.分子标记鉴别灰树花种质资源的研究[J].热带作物学报,2011,32(7):1330-1336.
[20]江玉姬,邓优锦,刘新锐,等.74株草菇遗传多样性分析[J].核农学报2015,29(11):2084-2092.
[21]韦仕岩,吴圣进,汪茜,等.草菇菌株的ISSR遗传差异分析[J].热带作物学报,2013,34(11):2209-2213.