基于nrDNA ITS对23种兜兰属植物的分子系统学分析
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摘要
采用PCR直接测序法,测定23种兜兰属植物的内部转录间隔区(ITS)序列,并结合GenBank中所查到的德氏兜兰(AJ564372)、波瓣兜兰(AY530916)及麻栗坡兜兰(AJ564357)的核糖体DNA(nrDNA)ITS区序列进行比对,确定23种兜兰的ITS1、5.8S及ITS2的界限。结果表明:23种兜兰植物的nrDNA ITS序列长度为717～787 bp;G+C含量为48.6%～51.5%,其中ITS1和ITS2长度分别为393～451 bp及155～180 bp,包括211个变异位点;109个信息位点。基于贝叶斯法和最大简约法构建系统聚类树,结果表明,2种方法构建的树基本一致,整个树分为2个分支3个亚组,表明基于nrDNA ITS序列能够鉴定分析兜兰属植物种间的亲缘关系。
引文
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[2]Guo Y Y,Huang L Q,Liu Z J,et al.Promise and challenge of DNA barcoding in Venus slipper (Paphiopedilum)[J].PLoS One,2016,11(1):1-13.
[3]Gorniak M,Szlachetko D L,Kowalkowska A K,et al.Taxonomic placement of Paphiopedilum canhii(Cypripedioideae;Orchidaceae) based on cytological,molecular and micromorphological evidence[J].Molecular Phylogenetics and Evolution,2014,70(1):429-441.
[4]黄云峰,薛跃规.中国新分布种海伦兜兰的濒危状况[J].植物分类学报,2007,45(3):333-336.
[5]罗毅波,贾建生,王春玲.中国兰科植物保育的现状和展望[J].生物多样性,2003,11(1):70-77.
[6]陈庆文,郭运青.中国兜兰属植物研究现状综述[J].广西农业科学,2010,41(8):818-821.
[7]Cribb P.The genus Paphiopedilum[M].2nd ed.National History Publications,1998:22-30.
[8]陈心启,刘方媛.云南几种兜兰属植物[J].云南植物研究,1982,4(2):163-167.
[9]Doyle J J.A rapid DNA isolation procedure for small amounts of fresh leaf tissue[J].Phytochem Bull,1987,19(1):11-15.
[10]Keyser M.DNAstar's lasergene genomics suite for RNA-Seq alignment and analysis of three Bos taurus breeds[J].Astronomische Nachrichten,2015,25(18):263-272.
[11]Tamura K,Peterson D,Peterson N,et al.MEGA 5:molecular evolutionary genetics analysis using maximum likelihood,evolutionary distance,and maximum parsimony methods[J].Molecular Biology and Evolution,2011,28(10):2731-2739.
[12]Huelsenbeck J P,Ronquist F.Mrbayes:Bayesian inference of phylogenetic trees[J].Bioinformatics,2001,17(8):754-755.
[13]Posada D.Jmodeltest:phylogenetic model averaging[J].Molecular Biology and Evolution,2008,25(7):1253-1256.
[14]Müller J,Müller K.TreeGraph:automated drawing of complex tree figures using an extensible tree description format[J].Molecular Ecology Notes,2004,4(4):786-788.
[15]Wang C,Zhang H,Qian Z Q,et al.Genetic differentiation in endangered Gynostemma pentaphyllum(Thunb.) Makino based on ISSR polymorphism and its implications for conservation[J].Biochemical Systematics and Ecology,2008,36(9):699-705.
[16]Baldwin B G,Sanderson M J,Porter J M,et al.The ITS region of nuclear ribosomal DNA:a valuable source of evidence on angiosperm phylogeny[J].Annals of the Missouri Botanical Garden,1995,82(2):247-277.
[17]Ainouche M L,Bayer R J.On the origins of the tetraploid Bromus species (section Bromus,Poaceae):insights from internal transcribed spacer sequences of nuclear ribosomal DNA[J].Genome,1997,40(5):730.
[18]Haffner E,Hellwing F H.Phylogeny of the tribe Cardueae (Compositae) with emphasis on the subtribe Carduinae:an analysis based on ITS sequence data[J].Willdenowia,1999,29(1/2):27-39.
[19]Donghue M J,Baldwin B G,LI J,et al.Viburnum phylogeny based on chloroplast trnK Intron and nuclear ribosomal ITS DNA sequences[J].Systematic Botany,2004,29(1):188-198.
[20]Graham S A,Freudenstein J V,Luker M.A phylogenetic study of Cuphea(Lythraceae) based on morphology and nuclear rDNA ITS sequences[J].Systematic Botany,2006,31(4):764-778.
[21]Chung S Y,Choi S H.Genetic variability and relationships among interspecific hybrid cultivars and parental species of Paphiopedilum via ribosomal DNA sequence analysis[J].Plant Systematics and Evolution,2012,298(10):1897-1907.
[22]孙彩云,张明永,叶秀粦.利用RAPD和同工酶研究中国兜兰属种间亲缘关系[J].园艺学报,2005,32(2):268-272.