魏潇;章秋平;刘威生;刘宁;刘硕;杨巍;

• 1:辽宁省果树科学研究所·国家果树种质熊岳李杏圃

摘要(Abstract)：

【目的】建立一种基于单碱基突变的双向等位基因特异PCR(bi-directional PCR amplification of specific alleles,Bi-PASA)技术,并将其应用于杏单核苷酸多态性分型研究。【方法】利用直接测序的方法在2份不同果肉质地杏EXP基因的DNA序列上发现1个单碱基变异。在其上、下游两端设计双向等位基因特异性引物及其外侧互补引物,对120份杏材料的SNP进行分型。同时,对部分材料的基因型进行测序验证。【结果】利用双向等位基因特异PCR对杏EXP基因315 bp处SNP分型结果与直接测序完全一致。【结论】双向等位基因特异PCR技术是一种简单、经济、快速而可靠的SNP分型方法,可有效地应用于杏基因组上已知SNP突变的分型研究。

关键词(KeyWords)： 杏;单核苷酸多态性;双向等位基因特异PCR

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(31071767);; 公益性行业(农业)科研专项(201003058)

作者(Author): 魏潇;章秋平;刘威生;刘宁;刘硕;杨巍;

Email:

DOI: 10.13925/j.cnki.gsxb.2013.06.013

参考文献(References)：

• [1]BROOKES A J.The essence of SNPs[J].Gene,1999,234:177-186.
• [2]YU X Q,BAI G H,LIU S W,LUO N,WANG Y,RICHMOND D S,PIJUT P M,JACKSON S A,YU J M,JIANG Y W.Association of candidate genes with drought tolerance traits in diverse perennial ryegrass accessions[J].Journal of Experimental Botany,2013,64(6):1537-1551.
• [3]GANAL M W,ALTMANN T,RODER M S.SNP identification in crop plants[J].Current Opinion in Plant Biology,2009,12(2):211-217.
• [4]CHEN Ji-bao,JING Rui-lian,YUAN Hai-yan,WEI Bo.Development of allele-specific PCR[J].Journal of Plant Genetic Resources,2006,6(4):469-473.陈吉宝,景蕊莲,员海燕,卫波.等位基因特异PCR技术的研究与应用[J].植物遗传资源学报,2006,6(4):469-473.
• [5]WANG Wei-peng,NI Kun-yi,ZHOU Guo-hua.Approaches for SNP genotyping[J].Hereditas,2006,28(1):117-126.汪维鹏,倪坤仪,周国华.单核苷酸多态性检测方法的研究进展[J].遗传,2006,28(1):117-126.
• [6]ALKAN C,COEL B P,EICHLER E E.Genome structural variation discovery and genotyping[J].Nature Reviews Genetics,2011,12:363-376.
• [7]YE S,DHILLON S,KE X,COLLINS A R,DAY I N M.An efficient procedure for genotyping single nucleotide polymorphisms[J].Nucle-ic Acids Research,2001,29(17):E88-8.
• [8]WANG Ke,ZHANG Jin-tao,YUN Yu-xia,WU Xiao-bing,CHEN A-qun,WANG Peng,WANG Kai-juan,ZHANG Jian-ying,DAI Li-ping.Improvement on PCR-CTPP:a SNP genotyping approach based on mismatch technique[J].Hereditas,2011,33(2):182-188.王柯,章金涛,运玉霞,吴晓冰,陈阿群,王鹏,王凯娟,张建营,代丽萍.PCR-CTPP:一种基于错配技术的SNP分型方法的改良[J].遗传,2011,33(2):182-188.
• [9]ALVES D M T,PEREIRA R W,LEAL-BERTIOLI S C M,MORETZSOHN M C,GUIMAR?ES P M,BERTIOLI D J.Development and use of single nucleotide polymorphism markers for candidate resistance genes in wild peanuts[J].Genetics and Molecular Research,2008,7(3):631-642.
• [10]CHIAPPARINO E,LEE D,DONINI P.Genotyping single nucleotide polymorphisms in barley by tetra-primer ARMS-PCR[J].Genome,2004,47:414-420.
• [11]HAYASHI K,HASHIMOTO N,DAIGEN M,ASHIKAWA I.Development of PCR-based SNP markers for rice blast resistance genes at the Piz locus[J].Theoretical and Applied Genetics,2004,108:1212-1220.