基于β-AS基因多态性的金铁锁核心种质构建
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摘要
金铁锁是"云南白药"的重要组分,为我国西南地区的特有二级濒危保护植物。其野生资源已不能满足医药生产的需求,亟需通过对其种质资源进行研究,构建金铁锁核心种质。为金铁锁种质改良、分子育种等奠定基础。该研究采用了关键酶基因(β-AS)作为核基因标记对金铁锁进行了谱系地理学研究。研究结果显示,在所有居群中,总共检测到36个单倍型,单倍型多样性(H_d=0.905)较高,居群间存在一定的遗传分化(G_(ST)=0.280)。AMOVA结果显示,金铁锁居群内遗传多样性较低(22.57%),居群间的遗传变异占77.43%。单倍型分布结合网状进化分析结果显示,Hap1为普通单倍型,且位于网状图的中心位置,可判断为祖先单倍型。Hap2,Hap4,Hap15,Hap16共享单倍型,剩余的为居群特有单倍型。在此基础上,为了最大限度的保存金铁锁的遗传多样性,依据药用植物核心种质构建的标准提出了金铁锁核心种质的构建策略。
Psammosilene tunicoides is one of the main ingredients of the "Yunnan Baiyao". P. tunicoides is an endangered species included in the secondary protection list in China Plant Red Data Book as well as the endemic species in Southwest China. Its natural resources could not meet the needs of pharmaceutical production. Construction of core collection of P. tunicoides will lay the foundation for germplasm improvement and molecular breeding. The sequence variation of the key enzymes gene locus(β-AS) were carried out to survey the population structure and population history of the species. Among the 11 populations across its geographical range, 36 haplotypes were identified. The levels of haplotype diversity(H_d=0.905) were high, while the levels of population differentiation(G_(ST)=0.280) were low. Analysisof molecular variance(AMOVA) indicated that a significantly greater proportion of total genetic variationpartitioned among populations thanwithin populations(values of 77.43% and 22.57%, respectively). These results in combination with the star-like phylogenetic network analysis indicate that Hap1 as an ancestral haplotypewas shared in four populations, Hap2, Hap4, Hap15 and Hap16 are occurred in two populations, the remains as private haplotype only distributed in single population. The strategy of core collection was constructed in order to maximumpreserve genetic diversity of P. tunicoides.
Psammosilene tunicoides is one of the main ingredients of the "Yunnan Baiyao". P. tunicoides is an endangered species included in the secondary protection list in China Plant Red Data Book as well as the endemic species in Southwest China. Its natural resources could not meet the needs of pharmaceutical production. Construction of core collection of P. tunicoides will lay the foundation for germplasm improvement and molecular breeding. The sequence variation of the key enzymes gene locus(β-AS) were carried out to survey the population structure and population history of the species. Among the 11 populations across its geographical range, 36 haplotypes were identified. The levels of haplotype diversity(H_d=0.905) were high, while the levels of population differentiation(G_(ST)=0.280) were low. Analysisof molecular variance(AMOVA) indicated that a significantly greater proportion of total genetic variationpartitioned among populations thanwithin populations(values of 77.43% and 22.57%, respectively). These results in combination with the star-like phylogenetic network analysis indicate that Hap1 as an ancestral haplotypewas shared in four populations, Hap2, Hap4, Hap15 and Hap16 are occurred in two populations, the remains as private haplotype only distributed in single population. The strategy of core collection was constructed in order to maximumpreserve genetic diversity of P. tunicoides.
引文
[1] 吴征镒.云南植物志.第6卷.种子植物[M]. 北京:科学出版社, 1995: 247.
[2] 傅立国. 中国植物红皮书:稀有濒危植物.第1册[M]. 北京: 科学出版社, 1992.
[3] 马小军, 肖培根. 种质资源遗传多样性在药用植物开发中的重要意义[J].中国中药杂志, 1998,23(10): 579.
[4] Frankel O H,Brown A H D. Current plant genetic resources-a critical appraisal[M]//Genetics: new frontiers.Vol.Ⅳ.New Delhi, India:Oxford and IBH Publishing, 1984: 1.
[5] 袁庆军, 黄璐琦, 郭兰萍,等. 展望分子谱系地理学在道地药材研究中的应用[J]. 中国中药杂志, 2009, 34(16): 2008.
[6] 黄璐琦. 分子生药学[M]. 2版.北京:北京大学医学出版社, 2006.
[7] 陈宏昊. 基于β-AS基因SNPs的甘草道地性及甘草酸快速积累机制研究[D]. 北京:北京中医药大学, 2014.
[8] 崔光红, 王学勇, 冯华,等. 丹参乙酰CoA酰基转移酶基因全长克隆和SNP分析[J]. 药学学报,2010,45(6):785.
[9] Librado P, Rozas J. DnaSP v5: a softwate for comprehensive analysis of DNA polymorphism data [J].Bioinformatics, 2009,25(11): 1451.
[10] Thompson J D, Gibson T J, Plewniak F, et al. The CLUSTAL_X windows interface: flexible strategies formultiple sequence alignment aided by quality analysis tools[J]. Nucleic Acids Res, 1997,25(24): 4876.
[11] Hall T A. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT [J]. Nucleic Acids Symp Ser,1999,41:95.
[12] Bandelt H J,Froster P, Rohe A. Median-joining networks for inferring intraspecific phylogenies [J]. Mol Biol Evol, 1999, 16(1):37.
[13] Dupanloup I, Schneider S, Excoffier L.A simulated annealing approach to deline the genetic structure of population[J]. Mol Ecol, 2002, 11(12):2571.
[14] Rousset F, Raymond M. Testing heterozygote excess and deficiency [J]. Genetics, 1995,140(4):1413.
[15] Excoffier L, Laval G, Schneider S. Arlequin ver. 3.1: an integrated software package for population genetics data analysis [J]. Evol Bioinform, 2005, 1:47.
[16] Petit R J, Elmousadik A, Pons O. Identifying populations for conservation on the basis of genetic markers[J]. Conserv Biol, 1998, 12(4):844.
[17] 赵保胜,桂海水,朱寅荻, 等.金铁锁化学成分、药理作用和临床应用研究进展[J].中国实验方剂学杂志,2011,17(18):288.
[18] 戴住波, 朱常成, 钱子刚, 等. 金铁锁种质资源的遗传多样性分析[J]. 中草药, 2007, 38(7): 1070.
[19] Qu Y, Yu H, Wu G, et al. Genetic diversity and population structure of the endangered species Psammosilene tunicoides revealed by DALP analysis[J]. Biochem Syst Ecol,2010, 38: 880.
[20] Zhang Q Y, Zhao Y J, Gong X.Genetic variation and phylogeography of Psammosilene tunicoides (Caryophyllaceae), a narrowly distributed and endemic species in south-western China[J].Aust J Bot, 2011, 59(5): 450.
[21] Wolfe K H, Li W H, Sharp P M. Rates of nucleotide substitution vary greatly among plant mitochondrial, chloroplast, and nuclear DNAs[J].Proc Natl Acad Sci USA, 1987, 84(24):9054.
[22] 刘晓光, 黄璐琦, 袁庆军, 等.基于分子谱系地理学的药用植物核心种质构建[J]. 中国中药杂志,2012,37(5): 695.
[2] 傅立国. 中国植物红皮书:稀有濒危植物.第1册[M]. 北京: 科学出版社, 1992.
[3] 马小军, 肖培根. 种质资源遗传多样性在药用植物开发中的重要意义[J].中国中药杂志, 1998,23(10): 579.
[4] Frankel O H,Brown A H D. Current plant genetic resources-a critical appraisal[M]//Genetics: new frontiers.Vol.Ⅳ.New Delhi, India:Oxford and IBH Publishing, 1984: 1.
[5] 袁庆军, 黄璐琦, 郭兰萍,等. 展望分子谱系地理学在道地药材研究中的应用[J]. 中国中药杂志, 2009, 34(16): 2008.
[6] 黄璐琦. 分子生药学[M]. 2版.北京:北京大学医学出版社, 2006.
[7] 陈宏昊. 基于β-AS基因SNPs的甘草道地性及甘草酸快速积累机制研究[D]. 北京:北京中医药大学, 2014.
[8] 崔光红, 王学勇, 冯华,等. 丹参乙酰CoA酰基转移酶基因全长克隆和SNP分析[J]. 药学学报,2010,45(6):785.
[9] Librado P, Rozas J. DnaSP v5: a softwate for comprehensive analysis of DNA polymorphism data [J].Bioinformatics, 2009,25(11): 1451.
[10] Thompson J D, Gibson T J, Plewniak F, et al. The CLUSTAL_X windows interface: flexible strategies formultiple sequence alignment aided by quality analysis tools[J]. Nucleic Acids Res, 1997,25(24): 4876.
[11] Hall T A. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT [J]. Nucleic Acids Symp Ser,1999,41:95.
[12] Bandelt H J,Froster P, Rohe A. Median-joining networks for inferring intraspecific phylogenies [J]. Mol Biol Evol, 1999, 16(1):37.
[13] Dupanloup I, Schneider S, Excoffier L.A simulated annealing approach to deline the genetic structure of population[J]. Mol Ecol, 2002, 11(12):2571.
[14] Rousset F, Raymond M. Testing heterozygote excess and deficiency [J]. Genetics, 1995,140(4):1413.
[15] Excoffier L, Laval G, Schneider S. Arlequin ver. 3.1: an integrated software package for population genetics data analysis [J]. Evol Bioinform, 2005, 1:47.
[16] Petit R J, Elmousadik A, Pons O. Identifying populations for conservation on the basis of genetic markers[J]. Conserv Biol, 1998, 12(4):844.
[17] 赵保胜,桂海水,朱寅荻, 等.金铁锁化学成分、药理作用和临床应用研究进展[J].中国实验方剂学杂志,2011,17(18):288.
[18] 戴住波, 朱常成, 钱子刚, 等. 金铁锁种质资源的遗传多样性分析[J]. 中草药, 2007, 38(7): 1070.
[19] Qu Y, Yu H, Wu G, et al. Genetic diversity and population structure of the endangered species Psammosilene tunicoides revealed by DALP analysis[J]. Biochem Syst Ecol,2010, 38: 880.
[20] Zhang Q Y, Zhao Y J, Gong X.Genetic variation and phylogeography of Psammosilene tunicoides (Caryophyllaceae), a narrowly distributed and endemic species in south-western China[J].Aust J Bot, 2011, 59(5): 450.
[21] Wolfe K H, Li W H, Sharp P M. Rates of nucleotide substitution vary greatly among plant mitochondrial, chloroplast, and nuclear DNAs[J].Proc Natl Acad Sci USA, 1987, 84(24):9054.
[22] 刘晓光, 黄璐琦, 袁庆军, 等.基于分子谱系地理学的药用植物核心种质构建[J]. 中国中药杂志,2012,37(5): 695.