基于SSR标记的山西晋南漆树种群遗传结构
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摘要
漆树(Toxicodendron vernicifluum (Stokes) F. A. Barkley)是中国重要的多年生落叶乔木,在生态群落保护和生物制药方面均有较高的利用价值,因此,具有重要的研究意义。本研究首先通过单因素试验确定了漆树SSR-PCR体系,进一步经正交试验对其SSR-PCR体系进行优化和筛选。通过PCR扩增筛选出7对条带清晰、多态性好的引物,7对引物最适退火温度分别是:57℃,55.6℃,58℃,57℃,56.8℃,56℃,58℃。基于建立的漆树SSR-PCR体系,利用选出的7对引物,对46个漆树样本进行遗传多样性及遗传结构分析,获得漆树种群内的观察等位基因平均数Na、有效等位基因数Ne、Shannon多样性指数I、Nei's基因多样性指数H分别为1.79、1.24、0.27、0.16,多态位点百分率PPB为88.72%,表明漆树种群内部具有丰富遗传多样性,AMOVA分子方差分析结果显示漆树种群内遗传变异74%高于种群间变异26%,表明种群内的遗传变异大于种群间。导致此种现象可能与漆树的传粉方式、种子特征和种子播撒机制以及环境抑制有一定的关系,本研究结果为合理开发利用漆树天然林,发掘其经济价值提供理论依据。
Toxicodendron vernicifluum(Stokes) F. A. Barkley, perennial deciduous plant, is an important economic and ecological tree in China. In this study, the SSR-PCR system of T. vernicifluum firstly made by single-factor experiment and be optimized, 7 pairs of primers with clear bands and good polymorphism were screened by PCR.And the optimal annealing temperature was 57℃, 55.6℃, 58℃, 57℃, 56.8℃, 56℃, 58℃. Based on the established SSR-PCR system, 7 pairs of primers with good polymorphism were selected to analyze the genetic diversity and genetic structure of 46 T. vernicifluum samples. The average number of alleles(Na), effective number of alleles(Ne), Shannon's information index(I), Nei's genetic diversity index(H) were respectively 1.79, 1.24, 0.27,0.16, percentage of polymorphic band(PPB) was 88.72%, It showed that the genetic diversity of T. vernicifluum population was rich. The results of AMOVA molecular variance analysis showed that the genetic variation in the population of T. vernicifluum(74%) was higher than that among the population(26%), which indicates that the genetic variation within the population is greater than that between populations. This phenomenon maybe relate to the pollination mode, seed characteristics, seed seeding mechanism and environmental inhibition of T. vernicifluum.The results of this study can provide a basis for the rational protection, development and utilization of natural T.vernicifluum forest and the economic value exploration.
Toxicodendron vernicifluum(Stokes) F. A. Barkley, perennial deciduous plant, is an important economic and ecological tree in China. In this study, the SSR-PCR system of T. vernicifluum firstly made by single-factor experiment and be optimized, 7 pairs of primers with clear bands and good polymorphism were screened by PCR.And the optimal annealing temperature was 57℃, 55.6℃, 58℃, 57℃, 56.8℃, 56℃, 58℃. Based on the established SSR-PCR system, 7 pairs of primers with good polymorphism were selected to analyze the genetic diversity and genetic structure of 46 T. vernicifluum samples. The average number of alleles(Na), effective number of alleles(Ne), Shannon's information index(I), Nei's genetic diversity index(H) were respectively 1.79, 1.24, 0.27,0.16, percentage of polymorphic band(PPB) was 88.72%, It showed that the genetic diversity of T. vernicifluum population was rich. The results of AMOVA molecular variance analysis showed that the genetic variation in the population of T. vernicifluum(74%) was higher than that among the population(26%), which indicates that the genetic variation within the population is greater than that between populations. This phenomenon maybe relate to the pollination mode, seed characteristics, seed seeding mechanism and environmental inhibition of T. vernicifluum.The results of this study can provide a basis for the rational protection, development and utilization of natural T.vernicifluum forest and the economic value exploration.
引文
Doyle J.J.,and Doyle J.L.,1987,A rapid DNA isolation procedure for small quantities of fresh leaf tissue,Phytochem.Bull.,19:11-15
Fischer M.,and Matthies D.,1998,Rapd variation in relation to population size and plant fitness in the rare Gentianella germanica(Gentianaceae),Am.J.Bot.,85(6):811-819
Ge X.J.,Zhang L.B.,and Yuan Y.M.,2005,Strong genetic differentiation of the East-Himal ayan Megacodon stylophorus(Gentianaceae)detected by Inter-Simple Sequence Repeats(ISSR),Biodivers.Conserv.,14(4):849-861
Guo C.Y.,Wang Z.L.,Wei P.F.,Su Y.,Li F.M.,and Chen Q.H.,2018,Cloning and expression analysis of FAD2 gene in Toxicodendron vernicifluum,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),16(19):6282-6291(郭崇炎,王志龙,韦鹏飞,苏莹,黎芳梅,陈秋红,2018,漆树TvFAD2基因的克隆与表达分析,分子植物育种,16(19):6282-6291)
Lai Y.,Jia J.L.,Wang J.M.,Ren L.,LüZ.Y.,Zhu H.Q.,Ma H.,Yang L.N.,and Li Z.R.,2017,Analysis of genetic diversity and association with agronomic traits in barley(Hordeum vulgare L.)introduced from abroad using SSR markers,Mailei Zuowu Xuebao(Journal of Triticeae Crops),37(2):197-204(赖勇,贾建磊,王晋民,任龙,吕仲昱,朱惠琴,马辉,杨莉娜,李宗仁,2017,外引大麦SSR标记遗传多样性及其与农艺性状的关联分析,麦类作物学报,37(2):197-204)
Li X.H.,Zhang H.,Wang D.Y.,Zhang L.,Shao J.W.,and Zhang X.P.,2013,The genetic structure of endemic plant Pteroceltis tatarinowii by ISSR markers,Shengtai Xuebao(Acta Ecologica Sinica),33(16):4892-4901(李晓红,张慧,王德元,张莉,邵剑文,张小平,2013,我国特有植物青檀遗传结构的ISSR分析,生态学报,33(16):4892-4901)
Nybom H.,and Bartish I.V.,2000,Effects of life history traits and sampling strategies on genetic diversity estimates obtained with RAPD markers in plants,Perspectives in Plant Ecology Evolution&Systematics,3(2):93-114
Slatkin M.,1987,Gene flow and the geographic structure of natural populations,Science,236(4803):787-792
Tong Z.J.,Chen X.J.,Fang D.H.,Zeng J.M.,Wu X.F.,and Xiao B.G.,2017,SSR marker-based analyses on genetic diversity and relevant variations of agronomic traits and chemical composition of 231 flue-cured tobacco germplasm resources,Zhongguo Yancao Xuebao(Acta Tabacaria Sinica),23(5):31-61(童治军,陈学军,方敦煌,曾建敏,吴兴富,肖炳光,2017,231份烤烟种质资源SSR标记遗传多样性及其与农艺性状和化学成分的关联分析,中国烟草学报,23(5):31-61)
Wei S.N.,Zhao X.P.,Tian M.J.,Li L.,and Hu Z.H.,2010,Application of plant morphology and AFLP molecular markers to identify Toxicodendron vernicifluum varieties of Shaanxi,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),30(4):665-671(魏朔南,赵喜萍,田敏爵,李林,胡正海,2010,应用植物形态学和AFLP分子标记鉴别陕西漆树品种,西北植物学报,30(4):665-671)
Wen Y.F.,Han W.J.,and Wu S.,2010,Plant genetic diversity and its influencing factors,Zhongnan Linye Keji Daxue Xuebao(Journal of Central South University of Forestry&Technology),30(12):80-87(文亚峰,韩文军,吴顺,2010,植物遗传多样性及其影响因素,中南林业科技大学学报,30(12):80-87)
Wright S.,1965,The interpretation of population structure by f-statistics with special regard to systems of mating,Evolution,19(6):395-420
Wu W.B.,He K.K.,Di H.,Niu S.H.,Ma Y.G.,Zhang Z.J.,and Li Y.,2018,Genetic structure and geographic system of geographical population of Pinus tabuliformis mountain range based on SSR in Shanxi Province of northern China,Beijing Linye Daxue Xuebao(Journal of Beijing Forestry University),40(10):51-59(武文斌,贺快快,狄皓,钮世辉,马彦光,张子杰,李悦,2018,基于SSR标记的山西省油松山脉地理种群遗传结构与地理系统,北京林业大学学报,40(10):51-59)
Zhang C.Q.,2015,Genetic diversity of population in Acer mono Maxim,Thesis for M.S.,Shanxi Normal University,Supervisor:Wang Y.L.,pp.22-28(张翠琴,2015,五角枫种群遗传多样性研究,硕士学位论文,山西师范大学,导师:王祎玲,pp.22-28)
Fischer M.,and Matthies D.,1998,Rapd variation in relation to population size and plant fitness in the rare Gentianella germanica(Gentianaceae),Am.J.Bot.,85(6):811-819
Ge X.J.,Zhang L.B.,and Yuan Y.M.,2005,Strong genetic differentiation of the East-Himal ayan Megacodon stylophorus(Gentianaceae)detected by Inter-Simple Sequence Repeats(ISSR),Biodivers.Conserv.,14(4):849-861
Guo C.Y.,Wang Z.L.,Wei P.F.,Su Y.,Li F.M.,and Chen Q.H.,2018,Cloning and expression analysis of FAD2 gene in Toxicodendron vernicifluum,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),16(19):6282-6291(郭崇炎,王志龙,韦鹏飞,苏莹,黎芳梅,陈秋红,2018,漆树TvFAD2基因的克隆与表达分析,分子植物育种,16(19):6282-6291)
Lai Y.,Jia J.L.,Wang J.M.,Ren L.,LüZ.Y.,Zhu H.Q.,Ma H.,Yang L.N.,and Li Z.R.,2017,Analysis of genetic diversity and association with agronomic traits in barley(Hordeum vulgare L.)introduced from abroad using SSR markers,Mailei Zuowu Xuebao(Journal of Triticeae Crops),37(2):197-204(赖勇,贾建磊,王晋民,任龙,吕仲昱,朱惠琴,马辉,杨莉娜,李宗仁,2017,外引大麦SSR标记遗传多样性及其与农艺性状的关联分析,麦类作物学报,37(2):197-204)
Li X.H.,Zhang H.,Wang D.Y.,Zhang L.,Shao J.W.,and Zhang X.P.,2013,The genetic structure of endemic plant Pteroceltis tatarinowii by ISSR markers,Shengtai Xuebao(Acta Ecologica Sinica),33(16):4892-4901(李晓红,张慧,王德元,张莉,邵剑文,张小平,2013,我国特有植物青檀遗传结构的ISSR分析,生态学报,33(16):4892-4901)
Nybom H.,and Bartish I.V.,2000,Effects of life history traits and sampling strategies on genetic diversity estimates obtained with RAPD markers in plants,Perspectives in Plant Ecology Evolution&Systematics,3(2):93-114
Slatkin M.,1987,Gene flow and the geographic structure of natural populations,Science,236(4803):787-792
Tong Z.J.,Chen X.J.,Fang D.H.,Zeng J.M.,Wu X.F.,and Xiao B.G.,2017,SSR marker-based analyses on genetic diversity and relevant variations of agronomic traits and chemical composition of 231 flue-cured tobacco germplasm resources,Zhongguo Yancao Xuebao(Acta Tabacaria Sinica),23(5):31-61(童治军,陈学军,方敦煌,曾建敏,吴兴富,肖炳光,2017,231份烤烟种质资源SSR标记遗传多样性及其与农艺性状和化学成分的关联分析,中国烟草学报,23(5):31-61)
Wei S.N.,Zhao X.P.,Tian M.J.,Li L.,and Hu Z.H.,2010,Application of plant morphology and AFLP molecular markers to identify Toxicodendron vernicifluum varieties of Shaanxi,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),30(4):665-671(魏朔南,赵喜萍,田敏爵,李林,胡正海,2010,应用植物形态学和AFLP分子标记鉴别陕西漆树品种,西北植物学报,30(4):665-671)
Wen Y.F.,Han W.J.,and Wu S.,2010,Plant genetic diversity and its influencing factors,Zhongnan Linye Keji Daxue Xuebao(Journal of Central South University of Forestry&Technology),30(12):80-87(文亚峰,韩文军,吴顺,2010,植物遗传多样性及其影响因素,中南林业科技大学学报,30(12):80-87)
Wright S.,1965,The interpretation of population structure by f-statistics with special regard to systems of mating,Evolution,19(6):395-420
Wu W.B.,He K.K.,Di H.,Niu S.H.,Ma Y.G.,Zhang Z.J.,and Li Y.,2018,Genetic structure and geographic system of geographical population of Pinus tabuliformis mountain range based on SSR in Shanxi Province of northern China,Beijing Linye Daxue Xuebao(Journal of Beijing Forestry University),40(10):51-59(武文斌,贺快快,狄皓,钮世辉,马彦光,张子杰,李悦,2018,基于SSR标记的山西省油松山脉地理种群遗传结构与地理系统,北京林业大学学报,40(10):51-59)
Zhang C.Q.,2015,Genetic diversity of population in Acer mono Maxim,Thesis for M.S.,Shanxi Normal University,Supervisor:Wang Y.L.,pp.22-28(张翠琴,2015,五角枫种群遗传多样性研究,硕士学位论文,山西师范大学,导师:王祎玲,pp.22-28)