云南白莺山茶树种质资源遗传多样性及群体结构分析
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摘要
为研究云南白莺山茶树种质资源的遗传多样性,以白莺山68份茶树种质为试验材料,进行生物学性状的调查,并采用EST-SSR分子标记毛细管电泳的方法对云南白莺山12个类型茶树遗传多样性、亲缘关系及群体遗传结构等内容进行研究。结果表明:(1)该群体生物学形态变异非常丰富,涵盖了乔木、小乔木和灌木;大叶种、中叶种和小叶种;叶形有椭圆形、长椭圆形和披针形;芽叶茸毛量呈现从无、少、中、多到特多的连续变化。(2)利用34对EST-SSR引物进行分子标记分析,表明云南白莺山群体期望杂合度平均值为0.6981,观测杂合度值为0.5982;PIC信息指数为0.6686,Shannon’s信息指数为1.6068,有效等位基因数和观测等位基因数分别为4.1761和9.7353。(3)基于Evanno统计模型的群体遗传结构分析可将白莺山茶树群体资源分为5个亚群,5个亚群间的基因流值为1.8995,群体间遗传分化系数(Fst)0.1163,表明各亚群是一个中度分化的群体,各亚群间存在着基因交流。
The paper aims to study the genetic diversity of tea germplasm in Baiying mountain. The capillary electrophoresis detection with fluorescent EST-SSR marker was used to analyze the genetic polymorphism and genetic relationship of 68 tea plant germplasm. The results showed that:(1) the population biology morphological variation was very rich, covering trees, small trees and shrubs; large leaf, middle leaf and small leaf; oval, oblong and lanceolate; the amount of bud fuzz changed continuously from less to more;(2) the 34 pairs of EST-SSR primers showed that the average of heterozygosity was 0.6981 in Baiying mountain, the observe heterozygosity was 0.5982, the polymorphism information content was 0.6686, the Shannon's information index was 1.6068; the number of digits and observed alleles were 4.1761 and 9.7353, respectively;(3) the tea germplasm was divided into 5 subpopulations based on Evanno statistical model, the gene flow value among five subpopulations was1.8995, and the coefficient of genetic differentiation between populations(Fst) was 0.1163. Each subgroup was a moderatelydifferentiatedgroup.Thereweregeneexchangesamong subgroups.
The paper aims to study the genetic diversity of tea germplasm in Baiying mountain. The capillary electrophoresis detection with fluorescent EST-SSR marker was used to analyze the genetic polymorphism and genetic relationship of 68 tea plant germplasm. The results showed that:(1) the population biology morphological variation was very rich, covering trees, small trees and shrubs; large leaf, middle leaf and small leaf; oval, oblong and lanceolate; the amount of bud fuzz changed continuously from less to more;(2) the 34 pairs of EST-SSR primers showed that the average of heterozygosity was 0.6981 in Baiying mountain, the observe heterozygosity was 0.5982, the polymorphism information content was 0.6686, the Shannon's information index was 1.6068; the number of digits and observed alleles were 4.1761 and 9.7353, respectively;(3) the tea germplasm was divided into 5 subpopulations based on Evanno statistical model, the gene flow value among five subpopulations was1.8995, and the coefficient of genetic differentiation between populations(Fst) was 0.1163. Each subgroup was a moderatelydifferentiatedgroup.Thereweregeneexchangesamong subgroups.
引文
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[9]Meegahakubura M K,Wambulwa M C,Thapa K.K,et al.Indications for ThteeIndenpent Domestication Events for the Tea Plant(Camellia sinensis(L)O.Kunte)and New Insights into the origin of Tea germplasm in China and India revealed by Nuclear Microsatellites[J].PLOS ONE,2016,11(5):e0155369.
[10]Ma J Q,Yao M Z,Ma C L,et al.Construction of a SSR-basedgenetic map and identification of QTLs for catechinscontentin tea plant(Camellia sinensis)[J].PLoS ONE,2014,9(3):e93131.
[11]Hu Chih-Yi,Lee Tair-Chyang,Tsai Hsien-Tsung,et al.Construction of an integrated genetic map based on maternal and paternal lineages of tea(Camellia sinensis)[J].Euphytica,2013,191:141-152.
[12]许鲲,吴金峰,李锋,等.2013-2015年中国冬油菜参试品种(系)DNA指纹数据库的初步构建及分析[J].中国油料作物学报,2017(2):160-169.
[13]陈文雄,季鹏章,黄兴奇,等.本山茶与勐库茶疑似杂交后代的RAPD鉴定[J].北方园艺,2008(10):153-155.
[14]毛娟,江鸿键,李崇兴,等.云南白莺山地区茶树遗传多样性研究[J].茶叶科学,2018,38(1):69-77.
[15]Wu Hualing,Chen Dong,Li Jiaxian,et al.De Novo Characterization of Leaf Transcriptome Using 454 Sequencing and Development of EST-SSR Markers in Tea[J].Plant MolBiol Rep,2013,31:524-538.
[16]Stewart C N Jr,Via L E.A rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications[J].Biotechniques,1993,14(5):748-750.
[17]Nei M.Estimation of average heterozygosity and genetic distance from a small number of individuals[J].Genetics,1978,89:583-590.
[18]Sewall Wright.Evolution and the Genetics of Populateons,Variability Within and Among Natural Populate ions[M].Chicago:The University of Chicago Press,1978:4.
[19]Evanno G,Regnaut S,Goudet J.Detecting the number of clusters of individuals using software STRUCTURE:A simulation study[J].Molecular Ecology,2005,14:2611-2620.
[20]黄晓霞,唐探,姜永雷,等.千家寨不同海拔野生茶树的EST-SSR遗传多样性研[J].茶叶科学,2015,35(4):347-353.
[2]张颖君,杨崇仁,曾恕芬,等.白莺山古茶的化学成分分析与栽培茶树的起源[J].云南植物研究,2010(1):77-82.
[3]吴华玲,李崇兴,陈栋,等.基于形态鉴定及叶片解剖结构对云南白莺山古茶树资源的聚类分析[J].中国农学通报,2014(31):114-121.
[4]Yao M Z,Ma C L,Qiao T T,et al.Diversity distribution and population structure of tea germplasms in China revealed by EST-SSR markers[J].Tree Genetics&Genomes,2012,8:205-220.
[5]Qiao T T,Ma C L,Zhou Y H,et al.EST-SSR genetic diversity and population structure of tea landraces anddeveloped cultivars(lines)in Zhejiang Province,China[J].Acta Agr Sinica,2010,36:744-753.
[6]李赛君,雷雨,段继华,等.基于EST-SSR的祁门种群体遗传多样性和亲缘关系分析[J].茶叶科学,2015,4:329-335.
[7]刘振,王新超,赵丽萍,等.基于EST-SSR的西南茶区茶树资源遗传多样性和亲缘关系分析[J].分子植物育种,2008,1:100-110.
[8]刘振,姚明哲,王新超,等.基于EST-SSR的福建地区茶树资源遗传多样性和亲缘关系分析[J].中国农业科学,2009,5:1720-1727.
[9]Meegahakubura M K,Wambulwa M C,Thapa K.K,et al.Indications for ThteeIndenpent Domestication Events for the Tea Plant(Camellia sinensis(L)O.Kunte)and New Insights into the origin of Tea germplasm in China and India revealed by Nuclear Microsatellites[J].PLOS ONE,2016,11(5):e0155369.
[10]Ma J Q,Yao M Z,Ma C L,et al.Construction of a SSR-basedgenetic map and identification of QTLs for catechinscontentin tea plant(Camellia sinensis)[J].PLoS ONE,2014,9(3):e93131.
[11]Hu Chih-Yi,Lee Tair-Chyang,Tsai Hsien-Tsung,et al.Construction of an integrated genetic map based on maternal and paternal lineages of tea(Camellia sinensis)[J].Euphytica,2013,191:141-152.
[12]许鲲,吴金峰,李锋,等.2013-2015年中国冬油菜参试品种(系)DNA指纹数据库的初步构建及分析[J].中国油料作物学报,2017(2):160-169.
[13]陈文雄,季鹏章,黄兴奇,等.本山茶与勐库茶疑似杂交后代的RAPD鉴定[J].北方园艺,2008(10):153-155.
[14]毛娟,江鸿键,李崇兴,等.云南白莺山地区茶树遗传多样性研究[J].茶叶科学,2018,38(1):69-77.
[15]Wu Hualing,Chen Dong,Li Jiaxian,et al.De Novo Characterization of Leaf Transcriptome Using 454 Sequencing and Development of EST-SSR Markers in Tea[J].Plant MolBiol Rep,2013,31:524-538.
[16]Stewart C N Jr,Via L E.A rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications[J].Biotechniques,1993,14(5):748-750.
[17]Nei M.Estimation of average heterozygosity and genetic distance from a small number of individuals[J].Genetics,1978,89:583-590.
[18]Sewall Wright.Evolution and the Genetics of Populateons,Variability Within and Among Natural Populate ions[M].Chicago:The University of Chicago Press,1978:4.
[19]Evanno G,Regnaut S,Goudet J.Detecting the number of clusters of individuals using software STRUCTURE:A simulation study[J].Molecular Ecology,2005,14:2611-2620.
[20]黄晓霞,唐探,姜永雷,等.千家寨不同海拔野生茶树的EST-SSR遗传多样性研[J].茶叶科学,2015,35(4):347-353.
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