甘薯IbANR基因的多态性分析
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摘要
花青素还原酶(ANR)是原花青素生物合成的关键酶,对甘薯色素的积累起重要作用.为了探讨甘薯花青素还原酶基因(IbANR)的多态性,根据徐紫薯3号cDNA序列设计特异引物,从27个不同的甘薯品种(品系)中扩增出IbANR基因序列,在编码区中发现72个变异位点,其中39个为SNP位点,检出频率为1SNP/488bp;内含子区变异出现的频率约为编码区的3倍.推测氨基酸序列也存在多态性.本研究为甘薯IbANR基因的功能研究及其在甘薯分子辅助育种中的应用提供支持.
Anthocyanidin reductase is a key enzyme in proanthocyanidins biosynthesis pathway.To explore the polymorphism of IbANRgene in sweetpotato,primers designed according to cDNA sequence of Xuzishu No.3 were uesd to amplify part DNA sequences of IbANRgene from 27 varieties of sweetpotato.The results of alignment and sequence analysis showed that total 72 mutate sites were found in the coding region.And among 72 mutations there were 39 SNPs with the frequency of 1 SNP/488 bp.The SNPs frequency in introns was about 3 times of that in coding region.Polymorphisms of putative amino acid were also found.These data might give support to function research of IbANRand application in molecular assistance breeding of sweetpotato.
Anthocyanidin reductase is a key enzyme in proanthocyanidins biosynthesis pathway.To explore the polymorphism of IbANRgene in sweetpotato,primers designed according to cDNA sequence of Xuzishu No.3 were uesd to amplify part DNA sequences of IbANRgene from 27 varieties of sweetpotato.The results of alignment and sequence analysis showed that total 72 mutate sites were found in the coding region.And among 72 mutations there were 39 SNPs with the frequency of 1 SNP/488 bp.The SNPs frequency in introns was about 3 times of that in coding region.Polymorphisms of putative amino acid were also found.These data might give support to function research of IbANRand application in molecular assistance breeding of sweetpotato.
引文
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[24]李雪,李志英,丛汉卿,等.红掌ANR基因克隆及其表达与佛焰苞的相观性分析[J].分子植物育种,2013,11(6):825.
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[33]毛新国,王爱萍,景蕊莲,等.小麦抗旱相关基因TaGSTF6的多态性[J].中国农业科学,2007,40(2):225.
[34]徐田军,陈松林.牙鲆MHC-DAA结构及等位基因多态性[J].遗传,2009,31(10):1120.
[35]Zhang X O,Wang H B,Zhang Y,et al.Complementary sequence-mediated exon circularization[J].Cell,2014,159(1):134.
[36]Gao M,Glorym A,Lakeisha S,et al.Wxintron variations support an allohexaploid origin of the sweetpotato[Ipomoea batatas(L.)Lam][J].Euphytica,2011,177(1):111.
[2]曹清河,刘义峰,李强,等.菜用甘薯国内外研究现状及展望[J].中国蔬菜,2007(10):41.
[3]Alain V,Denis M,Magali S C,et al.A review on SNP and othertypes of molecular markers and their use in animal genetics[J].Genet Sel Evol,2002,34(3):275.
[4]Brookes A J.The essence of SNPs[J].Gene,1999,234(2):177.
[5]杨昭庆,洪坤学,褚嘉佑.单核苷酸多态性的研究进展[J].国外医学:遗传学分册,2000,23(1):4.
[6]刘福平,白俊杰.单核苷酸多态性及其在水产动物遗传育种中的应用[J].中国水产科学,2008,15(4):704.
[7]康静婷,梁前进.单核苷酸多态性的研究及其生物学意义[J].生物学通报,2012,47(3):15.
[8]陈菁,孙桂芹.单核苷酸多态性在多基因遗传病研究中的作用[J].中国微生态学杂志,2007(2):229.
[9]张苏江,李建勇,施静艺,等.急性髓系白血病患者PDGFRβ和SHIP基因突变及其单核苷酸多态性研究[J].中华血液学杂志,2006,27(6):383.
[10]Tsui C,Coleman L E,Griffith J L,et al.Single nucleotide polymorphisms(SNPs)that map to gaps in the human SNP map[J].Nucleic Acids Res,2003,31(16):4910.
[11]Trjk O,Berger D,Meyer R C,et al.Establishment of a high-efficiency SNP-based framework marker set for Arabidopsis[J].Plant J,2003,36(1):122.
[12]Hayashi K,Hashimoto N,Daigen M,et al.Development of PCR-based SNP markers for rice blast resistance genes at the Piz locus[J].Theor Appl Genet,2004,108(7):1212.
[13]Rickert A M,Kim J H,Meyer S,et al.First-generation SNP/InDel markers tagging loci forpathogen resistance in the potato genome[J].Plant Biotechnol J,2003,1(6):399.
[14]陈吉宝,景蕊莲,员海燕,等.小麦TaDREB1基因的单核苷酸多态性分析[J].中国农业科学,2005,38(12):2387.
[15]He X Q,Liu Q C,Ishiki K,et al.Genetic variation and genetic relationships of Chinese sweetpotato landraces revealed by RAPD and AFLP markers[J].Breeding Sci,2006,56(2):201.
[16]He Xueqin,Liu Qingchang,Ishiki K,et al.ISSR analysis of genetic diversity and relationships among sweetpotato(Ipomoea batatas)landraces in China[J].Plant Genet Resour Newsl,2007,150:35.
[17]Zong X X,Redden R J,Liu Q C,et al.Analysis of a diverse global Pisumsp.collection and comparison to a Chinese local P.sativumcollection with microsatellite markers[J].Theor Appl Genet,2009,118(2):193.
[18]Wang L J,Dai C B,Liu D G,et al.Identification of a male-specific amplified fragment length polymorphism(AFLP)marker in Broussonetia papyrifera[J].African J Biotechnol,2012,11(33):8196.
[19]Liu D G,Zhao N,Zhai H,et al.AFLP fingerprinting and genetic diversity of main sweetpotato varieties in China[J].J Integrat Agricult,2012,11(9):1424.
[20]Erich G.The genetics and biochemistry of floral pigments[J].Ann Rev Plant Biol,2006,57(1):761.
[21]候夫云,王庆美,李爱贤,等.植物花青素合成酶的研究进展[J].中国农学通报,2009,25(2):188.
[22]Willians C A,Grayer R J.Anthocyanins and other flavonoids[J].Nat Prod Rep,2001,21(4):539.
[23]马敬,苏磊,袁美,等.花生C4H和ANR的克隆与表达分析[J].核农学报,2012,26(1):43.
[24]李雪,李志英,丛汉卿,等.红掌ANR基因克隆及其表达与佛焰苞的相观性分析[J].分子植物育种,2013,11(6):825.
[25]Murry M G,Thompson W F.Rapid isolation of high molecular weight plant DNA[J].Nucleic Acids Res,1980,8(19):4321.
[26]PBIL-IBCP-Lyon.Sopma secondary structure prediction method[ED/OL].[2016-09-10].http://npsapbil.ibcp.fr/cgi-bin/npsa automat.pl?page=/NPSA/npsa sopma.html.
[27]Shiotani I,Kawase T.Genomic structure of the sweet potato and hexaploid in Ipomoea trifida(H.B.K.)Don[J].Jpn J Breeding,1989,39(1):57.
[28]Hirakawa H,Okada Y,Tabuchi H,et al.Survey of genome sequences in a wild sweet potato,Ipomoea trifida(H.B.K.)G.Don[J].DNA Res,2015,22(2):171.
[29]Hawkins C,Caruana J,Schiksnis E,et al.Genomescale DNA variant analysis and functional validation of a SNP underlying yellow fruit color in wild strawberry[J].Sci Rep,2016(6):29017.
[30]Brookes A J.The essence of SNPs[J].Gene,1999,234(2):177.
[31]Guillet-Claude C,Birolleau-Touchard C,Manicacci D,et al.Nucleotide diversity of the ZmPox3 maize peroxidase gene:relationships between a MITE insertion in exon 2and variation in forage maize digestibility[J].BMC Genet,2004,5(1):1.
[32]Zhu Y L,Song Q J,Hyten D L,et al.Single-nucleotide polymorphisms in soybean[J].Genetics,2003,163(3):1123.
[33]毛新国,王爱萍,景蕊莲,等.小麦抗旱相关基因TaGSTF6的多态性[J].中国农业科学,2007,40(2):225.
[34]徐田军,陈松林.牙鲆MHC-DAA结构及等位基因多态性[J].遗传,2009,31(10):1120.
[35]Zhang X O,Wang H B,Zhang Y,et al.Complementary sequence-mediated exon circularization[J].Cell,2014,159(1):134.
[36]Gao M,Glorym A,Lakeisha S,et al.Wxintron variations support an allohexaploid origin of the sweetpotato[Ipomoea batatas(L.)Lam][J].Euphytica,2011,177(1):111.