基于SSR标记分析四川省桑树种质资源的群体结构和遗传多样性
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摘要
利用14对SSR引物对四川省89份桑树种质资源的基因组DNA进行PCR,分析种质资源间的群体结构和遗传多样性。共扩增出迁移率不同的条带65条,多态性条带65条,多态性条带比率达100%;Structure群体结构分析将89份桑树种质资源划分为4个群体,4个群体的多态性信息含量在0.552 0～0.599 8之间,总体为0.657 7;基因多样度在0.554 8～0.600 3之间,总体为0.579 0;等位基因数目在66～76个之间,总体为134个;等位基因丰富度在3.327 2～3.549 1之间,总体为3.983 1;遗传多样性主要来源于品种内,属中等偏高水平;14对SSR引物位点的总基因多样度Ht在0.291～0.931之间,整体为0.699;Nei's种群内基因多样度Hs为0.287～0.910,整体为0.662;种群间基因多样度为0.003～0.098,整体为0.037;种群间基因分化系数为0.008～0.047,整体为0.053;基因流为2.901～62.000,整体为8.934,遗传变异主要发生在群体内,群体间分化程度低,基因流大。UPGMA聚类分析与群体结构分析结果大致相同,2种分析结果都显示分群无地理迁移规律。
By using simple sequence repeat(SSR) molecular markers to amplify the genomic DNAs of 89 mulberry germplasm resources in Sichuan Province, the population structure and genetic diversity of these tested mulberry germplasm resources were analyzed. The result showed that amplification by 14 pairs of SSR primers yielded 65 bands, which showed polymorphism with a polymorphic percentage of 100%. Using software Structure, 89 mulberry germplasm resources were divided into 4 groups. The polymorphism information contents(PIC) of 4 groups were between 0.552 0 and 0.599 8, and the overall was 0.657 7. The gene diversity was between 0.554 8 and 0.600 3, 0.579 0 overall. The number of alleles was between 66 and 76, totally 134. The allelic richness of alleles was between 3.327 2 and 3.549 1, 3.983 1 overall. The genetic diversity was mainly from inter-varieties with a moderately high level. The total genetic diversity Ht of 14 SSR primer loci ranged from 0.291 to 0.931, 0.699 overall. The genetic diversity Hs in Nei's population was between 0.287 and 0.911, 0.662 overall. The genetic diversity among populations was from 0.003 to 0.098, 0.037 overall. The degree of gene differentiation among populations was from 0.008 to 0.047, 0.053 overall. The gene flow was from 2.901 to 62.000, 8.914 overall. The genetic variation mainly occurred within populations, while the degree of differentiation among groups was very low, and the gene flow was high. The results of UPGMA cluster analysis and population structure analysis are roughly the same, both of which show that there is no geographical migration rule.
By using simple sequence repeat(SSR) molecular markers to amplify the genomic DNAs of 89 mulberry germplasm resources in Sichuan Province, the population structure and genetic diversity of these tested mulberry germplasm resources were analyzed. The result showed that amplification by 14 pairs of SSR primers yielded 65 bands, which showed polymorphism with a polymorphic percentage of 100%. Using software Structure, 89 mulberry germplasm resources were divided into 4 groups. The polymorphism information contents(PIC) of 4 groups were between 0.552 0 and 0.599 8, and the overall was 0.657 7. The gene diversity was between 0.554 8 and 0.600 3, 0.579 0 overall. The number of alleles was between 66 and 76, totally 134. The allelic richness of alleles was between 3.327 2 and 3.549 1, 3.983 1 overall. The genetic diversity was mainly from inter-varieties with a moderately high level. The total genetic diversity Ht of 14 SSR primer loci ranged from 0.291 to 0.931, 0.699 overall. The genetic diversity Hs in Nei's population was between 0.287 and 0.911, 0.662 overall. The genetic diversity among populations was from 0.003 to 0.098, 0.037 overall. The degree of gene differentiation among populations was from 0.008 to 0.047, 0.053 overall. The gene flow was from 2.901 to 62.000, 8.914 overall. The genetic variation mainly occurred within populations, while the degree of differentiation among groups was very low, and the gene flow was high. The results of UPGMA cluster analysis and population structure analysis are roughly the same, both of which show that there is no geographical migration rule.
引文
[1] 陈仁芳.桑属系统学研究[D].武汉:华中农业大学,2010
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[13] 彭波,胡兴明,邓文,等.SSR标记与桑树发芽率、节间距性状的相关性研究[J].蚕业科学,2010,36(2):201-208
[14] 彭波,胡兴明,邓文,等.桑树种质资源SSR标记的遗传多样性分析[J].湖北农业科学,2010,49(4):779-784
[15] 高丽霞.桑树EST-SSR引物开发[J].南方农业学报,2013,44(8):1254-1257
[16] 罗义维,亓希武,帅琴,等.桑树杂交组合亲本的SSR标记多态性及遗传背景分析[J].蚕业科学,2014,40(4):576-581
[17] 姚刘慧,唐翠明,戴凡炜,等.基于桑树转录组测序的SSR标记开发和引物筛选[J].蚕业科学,2015,41(1):18-27
[18] WEISING K H,WOLFF K,MEYER W.DNA fingerprinting in plants and fungi[M].Boca Raton:CRC Press,1995:50-54
[19] 赵卫国.桑种质资源的遗传多样性及分子系统学研究[D].北京:中国农业科学院研究生院,2005
[20] PRITCHARD J K,STEPHENS M,DONNELLY P.Inference of population structure using multilocus genotype data[J].Genetics,2000,155(2):945-959
[21] YEH F C,YANG R C,BOYLE T.POPGENE:microsoft windows-based freeware for population genetic analysis release 1.31[M].Edmonton:University of Alberta,1999:1-28
[22] GOUDET J.FSTAT,a program to estimate and test gene diversity and fixation indices (version 2.9.3)[CP].http://www2.unil.ch/popgen/softwares/fstat.htm
[23] ROHLF F J.NTSYS-pc 2.1:numerical taxonomy and multivariate analysis system version 2.1[M].New York:Exeter Publishing,1988:75
[24] BOTSTEIN D,WHITE R L,SKOLNICK M,et al.Construction of a genetic linkage map in man using restriction fragment length polymorphisms[J].Am J Hum Genet,1980,32(3):314-331
[25] 中国科学院中国植物志编辑委员会.中国植物志[M].北京:科学出版社,1998:17-18
[26] 中国农业科学院蚕业研究所.中国桑树栽培学[M].上海:上海科学技术出版社,1985:46-49
[27] 刘凯旋,陈仁芳,刘玲,等.滇桑Morus yunnanensis Koidz.的特征、分布与系统位置[J].四川蚕业,2015,43(3):43-44
[2] 何大彦,朱万民.四川桑树品种[M].成都:四川科学技术出版社,1988:1-42
[3] ZIETKIEWICZ E,RAFAISKI A,LABUDA D.Genome fingerprinting by simple sequence repeat(SSR) anchored polymerase chain reaction amplification[J].Genomics,1994,20(2):176-183
[4] 郭瑞星,刘小红,荣廷昭,等.植物SSR标记的发展及其在遗传育种中的应用[J].玉米科学,2005,13(2):8-11
[5] AGGARWAL R K,UDAYKUMAR D,HENDRE P S,et al.Isolation and characterization of six novel microsatellite markers for mulberry (Morus indica)[J].Mol Ecol Resour,2010,4(3):477-479
[6] VENKATESWARLU M,URS S R,NATH B S,et al.A first genetic linkage map of mulberry (Morus,spp.) using RAPD,ISSR,and SSR markers and pseudotestcross mapping strategy[J].Tree Genet Genom,2006,3(1):15-24
[7] MATHITHUMILAN B,KADAM N N,BIRADAR J,et al.Development and characterization of microsatellite markers for Morus spp.and assessment of their transferability to other closely related species[J/OL].BMC Plant Biol,2013,13:194[2018-04-25].https://www.ncbi.nlm.nih.gov/pubmed/24289047.DOI:10.1186/1471-2229-13-194
[8] KRISHNAN R R,SUMATHY R,BINDROO B B,et al.MulSatDB:a first online database for mulberry microsatellites[J].Trees,2014,28(6):1793-1799
[9] KRISHNAN R R,NAIK V G,RAMESH S R,et al.Microsatellite marker analysis reveals the events of the introduction and spread of cultivated mulberry in the Indian subcontinent[J].Plant Genet Resour,2014,12(1):129-139
[10] ZHAO W G,MIAO X X,JIA S H,et al.Isolation and characterization of microsatellite loci from the mulberry,Morus L.[J].Plant,2005,168(2):519-525
[11] ZHAO W G,ZHOU Z H,MIAO X X,et al.A comparison of genetic variation among wild and cultivated Morus Species (Moraceae:Morus) as revealed by ISSR and SSR markers[J].Biodivers Conserv,2007,16(2):275-290
[12] 彭波,胡兴明,邓文,等.SSR标记与桑树表型性状的关联分析[C]//广西壮族自治区蚕业技术推广总站.中南五省区蚕桑育种协作研讨会论文集.武汉:武汉大学生命科学学院,2009:51-55
[13] 彭波,胡兴明,邓文,等.SSR标记与桑树发芽率、节间距性状的相关性研究[J].蚕业科学,2010,36(2):201-208
[14] 彭波,胡兴明,邓文,等.桑树种质资源SSR标记的遗传多样性分析[J].湖北农业科学,2010,49(4):779-784
[15] 高丽霞.桑树EST-SSR引物开发[J].南方农业学报,2013,44(8):1254-1257
[16] 罗义维,亓希武,帅琴,等.桑树杂交组合亲本的SSR标记多态性及遗传背景分析[J].蚕业科学,2014,40(4):576-581
[17] 姚刘慧,唐翠明,戴凡炜,等.基于桑树转录组测序的SSR标记开发和引物筛选[J].蚕业科学,2015,41(1):18-27
[18] WEISING K H,WOLFF K,MEYER W.DNA fingerprinting in plants and fungi[M].Boca Raton:CRC Press,1995:50-54
[19] 赵卫国.桑种质资源的遗传多样性及分子系统学研究[D].北京:中国农业科学院研究生院,2005
[20] PRITCHARD J K,STEPHENS M,DONNELLY P.Inference of population structure using multilocus genotype data[J].Genetics,2000,155(2):945-959
[21] YEH F C,YANG R C,BOYLE T.POPGENE:microsoft windows-based freeware for population genetic analysis release 1.31[M].Edmonton:University of Alberta,1999:1-28
[22] GOUDET J.FSTAT,a program to estimate and test gene diversity and fixation indices (version 2.9.3)[CP].http://www2.unil.ch/popgen/softwares/fstat.htm
[23] ROHLF F J.NTSYS-pc 2.1:numerical taxonomy and multivariate analysis system version 2.1[M].New York:Exeter Publishing,1988:75
[24] BOTSTEIN D,WHITE R L,SKOLNICK M,et al.Construction of a genetic linkage map in man using restriction fragment length polymorphisms[J].Am J Hum Genet,1980,32(3):314-331
[25] 中国科学院中国植物志编辑委员会.中国植物志[M].北京:科学出版社,1998:17-18
[26] 中国农业科学院蚕业研究所.中国桑树栽培学[M].上海:上海科学技术出版社,1985:46-49
[27] 刘凯旋,陈仁芳,刘玲,等.滇桑Morus yunnanensis Koidz.的特征、分布与系统位置[J].四川蚕业,2015,43(3):43-44