金花茶边缘种群及变种小果金花茶的遗传多样性分析
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摘要
利用10对SSR引物对南宁地区5个金花茶种群进行遗传多样性分析,探讨金花茶边缘种群及变种小果金花茶的遗传多样性,为金花茶的合理保护提供科学依据。结果显示:小果金花茶平均等位基因数(A)、有效等位基因数(Ae)、期望杂合度(He)和观测杂合度(Ho)分别为4.40、2.433、0.533和0.429,表明小果金花茶具有较低水平的遗传多样性。STRUCTURE聚类分析显示,5个金花茶种群按照两变种分为2个组。AMOVA结果和遗传分化系数(FST)均表明两变种间存在很大的遗传分化。位于平果县坡造乡的金花茶边缘种群具有最低水平的遗传多样性(PPB=50%, A=1.70, Ho=0.100),与主分布区种群间存在很大的遗传分化。基于以上结果表明,小果金花茶和平果县坡造乡边缘种群应当作为独立保护单元保护起来。
In ord er to provide a scientific basis for the reasonable conservation of Camellia nitidissima, we analyzed the genetic diversity of peripheral population C. nitidissima and the variety microcarpa from 5 Camellia nitidissima populations in Nanning area by using 10 pairs of SSR primers. The results showed that the average allele number(A), effective number of alleles(Ae), expected heterozygosity(He) and observed heterozygosity(Ho)were 4.40, 2.433, 0.533 and 0.429, respectively, indicating that the variety microcarpa had relatively low genetic diversity. STRUCTURE clustering analysis indicated that 5 C. nitidissima populations formed two groups according to two varieties. AMOVA result and genetic differentiation coefficient FSTvalue both revealed high genetic differentiation between two varieties. Peripheral population of C. nitidissima which located in Pozao town, Pingguo county had the lowest genetic diversity(PPB=50%, A=1.70, Ho=0.100), and high genetic differentiation between peripheral population and the main distribution area of C. nitidissima. Based on the above results, C. nitidissima var. microcarpa and peripheral population in Pozao town, Pingguo county should be recognized as independent conservation units.
In ord er to provide a scientific basis for the reasonable conservation of Camellia nitidissima, we analyzed the genetic diversity of peripheral population C. nitidissima and the variety microcarpa from 5 Camellia nitidissima populations in Nanning area by using 10 pairs of SSR primers. The results showed that the average allele number(A), effective number of alleles(Ae), expected heterozygosity(He) and observed heterozygosity(Ho)were 4.40, 2.433, 0.533 and 0.429, respectively, indicating that the variety microcarpa had relatively low genetic diversity. STRUCTURE clustering analysis indicated that 5 C. nitidissima populations formed two groups according to two varieties. AMOVA result and genetic differentiation coefficient FSTvalue both revealed high genetic differentiation between two varieties. Peripheral population of C. nitidissima which located in Pozao town, Pingguo county had the lowest genetic diversity(PPB=50%, A=1.70, Ho=0.100), and high genetic differentiation between peripheral population and the main distribution area of C. nitidissima. Based on the above results, C. nitidissima var. microcarpa and peripheral population in Pozao town, Pingguo county should be recognized as independent conservation units.
引文
Bin X.Y.,Tang S.Q.,Zhou J.Y.,Song H.T.,and Li Z.Y.,2005,ISSR analysis on genetic diversity of Camellia nitidissima Chi(Theaceae)in China,Wuhan Zhiwuxue Yanjiu(Journal of Wuhan Botanical Research),23(1):20-26(宾晓芸,唐绍清,周俊亚,宋洪涛,黎尊谊,2005,金花茶遗传多样性的ISSR分析,武汉植物学研究,23(1):20-26)
Chen X.Y.,Lu H.P.,Shen L.,and Li Y.Y.,2002,Identifying populations for priority conservation of important species,Shengwu Duoyangxing(Biodiversity Science),10(3):332-338(陈小勇,陆慧萍,沈浪,李媛媛,2002,重要物种优先保护种群的确定,生物多样性,10(3):332-338)
Doyle J.,ed.,1991,DNA protocols for plants-CTAB total DNAisolation,Springer,Berlin,Germany,pp.283-293
Eckert C.G.,Samis K.E.,and Lougheed S.C.,2008,Genetic variation across species'geographical ranges:the central-marginal hypothesis and beyond,Mol.Ecol.,17(5):1170-1188
Excoffier L.,Laval G.,and Schneider S.,2005,Arlequin(version3.0):an integrated software package for population genetics data analysis,Evol.Bioinform.Online,1:47-50
Frankham R.,Ballou J.D.,and Briscoe D.A.,eds.,Huang H.W.,and Kang M.,trans.,2004,Introduction to conservation genetics,Science Press,Beijing,China,pp.29-30(法兰克汉R.巴卢J.D.,布里斯科D.A.,主编,黄宏文,康明,主译,2004保育遗传学,科学出版社,中国,北京,pp.29-30)
He D.Y.,Li X.Y.,Wang L.L.,Zhang P.,Li S.Y.,and Xu Y.P.2016,Chemical constituents and pharmacological effects of Camellia nitidissima,Zhongguo Shiyan Fangjixue Zazhi(C-hinese Journal of Experimental Traditional Medical Formulae),22(3):231-234(贺栋业,李晓宇,王丽丽,张萍,李淑英,徐永平,2016,金花茶化学成分及药理作用研究进展中国实验方剂学杂志,22(3):231-234)
Huang R.B.,He T.P.,Zhuang J.,and Pang J.,2007,Plant resource and it's conservational countermeasure of Sect.Chrysantha Chang in Fangchenggang,Guangxi Nongye Shengwu Kexue(Journal of Guangxi Agricultural and Biological Science),26(S):32-37(黄瑞斌,和太平,庄嘉,庞洁,2007,广西防城港市金花茶组植物资源及其保育对策,广西农业生物科学,26(S):32-37)
Jakobsson M.,and Rosenberg N.A.,2007,CLUMPP:a cluster matching and permutation program for dealing with labe switching and multimodality in analysis of population structure,Bioinformatics,23(14):1801-1806
Li A.,and Ge S.,2002,Advances in plant conservation genetics Shengwu Duoyangxing(Biodiversity Science),10(1):61-71(李昂,葛颂,2002,植物保护遗传学研究进展,生物多样性10(1):61-71)
Liufu Y.Q.,Peng G.Q.,Lu Y.B.,Ye Q.Q.,and Tang S.Q.,2014Development and characterization of 38 microsatellite markers for Camellia flavida based on transcriptome sequencing,Conserv.Genet.Resour.,6(4):1007-1010
Lu Y.B.,2015,The population genetic structure of Camellia flavida Chang,Thesis for M.S.,College of Life Science,Guangxi Normal University,Supervisor:Tang S.Q.,pp.21-22(卢永彬,2015,淡黄金花茶种群遗传结构研究,硕士学位论文,广西师范大学生命科学学院,导师:唐绍清,pp.21-22)
Lu Y.B.,Liufu Y.Q.,Peng G.Q.,Ye Q.Q.,and Tang S.Q.,2014Development of 21 microsatellite primers for Camellia pingguoensis(Theaceae)using 454 sequencing,Conserv.Genet Resour.,6(3):791-793
Ming T.L.,and Bartholomew B.,2007,Theaceae,In:Editoria Committee(ed.),Flora of China,Science Press,Beijing,China,pp.368-372
Peakall R.,and Smouse P.E.,2012,GenAlEx 6.5:genetic analysis in Excel,Population genetic software for teaching and research-an update,Bioinformatics,28(19):2537-2539
Pritchard J.K.,Stephens M.,and Donnelly P.,2000,Inference of population structure using multilocus genotype data,Genetics,155(2):945-959
Qin H.N.,Yang Y.,Dong S.Y.,He Q.,Jia Y.,Zhao L.N.,Yu S.X.,Liu H.Y.,Liu B.,Yan Y.H.,Xiang J.Y.,Xia N.H.,Peng H.,Li Z.Y.,Zhang Z.X.,He X.J.,Yin L.K.,Lin Y.L.,Liu Q.R.,Hou Y.T.,Liu Y.,Liu Q.X.,Cao W.,Li J.Q.,Chen S.L.,Jin X.H.,Gao T.G.,Chen W.L.,Ma H.Y.,Geng Y.Y.,Jin X.F.,Chang Z.Y.,Jiang H.,Cai L.,Zang C.X.,Wu J.Y.,Ye J.F.,Lai Y.J.,Liu B.,Lin Q.W.,and Xue N.X.,2017,Threatened species list of China's higher plants,Shengwu Duoyangxing(Biodiversity Science),25(7):696-744(覃海宁,杨永,董仕勇,何强,贾渝,赵莉娜,于胜祥,刘慧圆,刘博,严岳鸿,向建英,夏念和,彭华,李振宇,张志翔,何兴金,尹林克,林余霖,刘全儒,侯元同,刘演,刘启新,曹伟,李建强,陈世龙,金效华,高天刚陈文俐,马海英,耿玉英,金孝锋,常朝阳,蒋宏,蔡蕾,臧春鑫,武建勇,叶建飞,赖阳均,刘冰,林秦文,薛纳新,2017,中国高等植物受威胁物种名录,生物多样性,25(7):696-744)
Rice W.R.,1989,Analyzing tables of statistical tests,Evolution,43(1):223-225
Rosenberg N.A.,2004,DISTRUCT:a program for the graphical display of population structure,Mol.Ecol.Notes,4(1):137-138
Rousset F.,2008,Genepop'007:a complete re-implementation of the genepop software for Windows and Linux,Mol.Ecol.Resour.,8(1):103-106
Su Z.M.,and Mo X.L.,1988,Geographic distribution of Camellia section Chrysantha from China,Guangxi Zhiwu(Guihaia),8(1):75-81(苏宗明,莫新礼,1988,我国金花茶组植物的地理分布,广西植物,8(1):75-81)
Tang S.Q.,Bin X.Y.,Wang L.,and Zhong Y.,2006,Genetic diversity and population structure of yellow Camellia(Camellia nitidissima)in China as revealed by RAPD and AFLPmarkers,Biochem.Genet.,44(9-10):449-461
Wei X.,Cao H.L.,Jiang Y.S.,Ye W.H.,Ge X.J.,and Li F.,2008,Population genetic structure of Camellia nitidissima(Theaceae)and conservation implications,Bot.Stud.,49(2):147-153
Wright S.,1931,Evolution in mendelian populations,Genetics,16(2):97-159
Chen X.Y.,Lu H.P.,Shen L.,and Li Y.Y.,2002,Identifying populations for priority conservation of important species,Shengwu Duoyangxing(Biodiversity Science),10(3):332-338(陈小勇,陆慧萍,沈浪,李媛媛,2002,重要物种优先保护种群的确定,生物多样性,10(3):332-338)
Doyle J.,ed.,1991,DNA protocols for plants-CTAB total DNAisolation,Springer,Berlin,Germany,pp.283-293
Eckert C.G.,Samis K.E.,and Lougheed S.C.,2008,Genetic variation across species'geographical ranges:the central-marginal hypothesis and beyond,Mol.Ecol.,17(5):1170-1188
Excoffier L.,Laval G.,and Schneider S.,2005,Arlequin(version3.0):an integrated software package for population genetics data analysis,Evol.Bioinform.Online,1:47-50
Frankham R.,Ballou J.D.,and Briscoe D.A.,eds.,Huang H.W.,and Kang M.,trans.,2004,Introduction to conservation genetics,Science Press,Beijing,China,pp.29-30(法兰克汉R.巴卢J.D.,布里斯科D.A.,主编,黄宏文,康明,主译,2004保育遗传学,科学出版社,中国,北京,pp.29-30)
He D.Y.,Li X.Y.,Wang L.L.,Zhang P.,Li S.Y.,and Xu Y.P.2016,Chemical constituents and pharmacological effects of Camellia nitidissima,Zhongguo Shiyan Fangjixue Zazhi(C-hinese Journal of Experimental Traditional Medical Formulae),22(3):231-234(贺栋业,李晓宇,王丽丽,张萍,李淑英,徐永平,2016,金花茶化学成分及药理作用研究进展中国实验方剂学杂志,22(3):231-234)
Huang R.B.,He T.P.,Zhuang J.,and Pang J.,2007,Plant resource and it's conservational countermeasure of Sect.Chrysantha Chang in Fangchenggang,Guangxi Nongye Shengwu Kexue(Journal of Guangxi Agricultural and Biological Science),26(S):32-37(黄瑞斌,和太平,庄嘉,庞洁,2007,广西防城港市金花茶组植物资源及其保育对策,广西农业生物科学,26(S):32-37)
Jakobsson M.,and Rosenberg N.A.,2007,CLUMPP:a cluster matching and permutation program for dealing with labe switching and multimodality in analysis of population structure,Bioinformatics,23(14):1801-1806
Li A.,and Ge S.,2002,Advances in plant conservation genetics Shengwu Duoyangxing(Biodiversity Science),10(1):61-71(李昂,葛颂,2002,植物保护遗传学研究进展,生物多样性10(1):61-71)
Liufu Y.Q.,Peng G.Q.,Lu Y.B.,Ye Q.Q.,and Tang S.Q.,2014Development and characterization of 38 microsatellite markers for Camellia flavida based on transcriptome sequencing,Conserv.Genet.Resour.,6(4):1007-1010
Lu Y.B.,2015,The population genetic structure of Camellia flavida Chang,Thesis for M.S.,College of Life Science,Guangxi Normal University,Supervisor:Tang S.Q.,pp.21-22(卢永彬,2015,淡黄金花茶种群遗传结构研究,硕士学位论文,广西师范大学生命科学学院,导师:唐绍清,pp.21-22)
Lu Y.B.,Liufu Y.Q.,Peng G.Q.,Ye Q.Q.,and Tang S.Q.,2014Development of 21 microsatellite primers for Camellia pingguoensis(Theaceae)using 454 sequencing,Conserv.Genet Resour.,6(3):791-793
Ming T.L.,and Bartholomew B.,2007,Theaceae,In:Editoria Committee(ed.),Flora of China,Science Press,Beijing,China,pp.368-372
Peakall R.,and Smouse P.E.,2012,GenAlEx 6.5:genetic analysis in Excel,Population genetic software for teaching and research-an update,Bioinformatics,28(19):2537-2539
Pritchard J.K.,Stephens M.,and Donnelly P.,2000,Inference of population structure using multilocus genotype data,Genetics,155(2):945-959
Qin H.N.,Yang Y.,Dong S.Y.,He Q.,Jia Y.,Zhao L.N.,Yu S.X.,Liu H.Y.,Liu B.,Yan Y.H.,Xiang J.Y.,Xia N.H.,Peng H.,Li Z.Y.,Zhang Z.X.,He X.J.,Yin L.K.,Lin Y.L.,Liu Q.R.,Hou Y.T.,Liu Y.,Liu Q.X.,Cao W.,Li J.Q.,Chen S.L.,Jin X.H.,Gao T.G.,Chen W.L.,Ma H.Y.,Geng Y.Y.,Jin X.F.,Chang Z.Y.,Jiang H.,Cai L.,Zang C.X.,Wu J.Y.,Ye J.F.,Lai Y.J.,Liu B.,Lin Q.W.,and Xue N.X.,2017,Threatened species list of China's higher plants,Shengwu Duoyangxing(Biodiversity Science),25(7):696-744(覃海宁,杨永,董仕勇,何强,贾渝,赵莉娜,于胜祥,刘慧圆,刘博,严岳鸿,向建英,夏念和,彭华,李振宇,张志翔,何兴金,尹林克,林余霖,刘全儒,侯元同,刘演,刘启新,曹伟,李建强,陈世龙,金效华,高天刚陈文俐,马海英,耿玉英,金孝锋,常朝阳,蒋宏,蔡蕾,臧春鑫,武建勇,叶建飞,赖阳均,刘冰,林秦文,薛纳新,2017,中国高等植物受威胁物种名录,生物多样性,25(7):696-744)
Rice W.R.,1989,Analyzing tables of statistical tests,Evolution,43(1):223-225
Rosenberg N.A.,2004,DISTRUCT:a program for the graphical display of population structure,Mol.Ecol.Notes,4(1):137-138
Rousset F.,2008,Genepop'007:a complete re-implementation of the genepop software for Windows and Linux,Mol.Ecol.Resour.,8(1):103-106
Su Z.M.,and Mo X.L.,1988,Geographic distribution of Camellia section Chrysantha from China,Guangxi Zhiwu(Guihaia),8(1):75-81(苏宗明,莫新礼,1988,我国金花茶组植物的地理分布,广西植物,8(1):75-81)
Tang S.Q.,Bin X.Y.,Wang L.,and Zhong Y.,2006,Genetic diversity and population structure of yellow Camellia(Camellia nitidissima)in China as revealed by RAPD and AFLPmarkers,Biochem.Genet.,44(9-10):449-461
Wei X.,Cao H.L.,Jiang Y.S.,Ye W.H.,Ge X.J.,and Li F.,2008,Population genetic structure of Camellia nitidissima(Theaceae)and conservation implications,Bot.Stud.,49(2):147-153
Wright S.,1931,Evolution in mendelian populations,Genetics,16(2):97-159