猕猴桃DNA条形码标记的筛选  

Screening of DNA Barcode for Actinidia

作　　者：叶婵娟 杨妙贤[1] 刘文[1,2] 梁红[1] 

机构地区：[1]仲恺农业工程学院生命科学学院,广东广州510225 [2]华南农业大学农学院,广东广州510642

出　　处：《北方园艺》2015年第24期79-83,共5页Northern Horticulture

基　　金：广东省教育厅科技攻关重点资助项目(粤财教[2011]473号);广东省科技计划资助项目(2013B090900012)

摘　　要：以5种野生猕猴桃为试验材料,采用DNA条形码技术,对6个DNA条形码标记rbcL、matK、psbA、trnL-F、ITS、trnH-psbA进行序列分析,筛选出能鉴别猕猴桃种间分子差异的DNA条形码,以期利用DNA条形码技术用于猕猴桃育种。结果表明：通过序列碱基含量分析,trnL-F和ITS条形码序列的GC含量最高,为51.1%～55.2%,trnH-psbA的GC含量最小,约为32.0%;通过Blast比对,结果显示5个猕猴桃野生种与基因库中已登录种类的6个DNA条形码相似度在99%以上,而有些与基因库中已登录种类有所差异,体现出猕猴桃种内的遗传多样性或种间存在基因渗透;从系统进化树结果分析显示,ITS标记能将上述5个野生种明显区分,基因进化多样性两两比对分析显示,5个猕猴桃野生种之间差异明显;Tajima’s中性检验中,trnL-F,ITS和matK具有较高的核苷酸多样性和中性检验值。研究比较了6种DNA条形码标记,认为ITS种间多样性较高,差异明显,较适宜作为猕猴桃DNA条形码。DNA barcode technology could used for the kiwifruit breeding usually.In this study,six DNA barcode,rbcL,matK,psbA,trnL-F,ITSand trnH-psbA were used to identify five wild species of Actinidia.The results showed that,GC contents of trnL-Fand ITS were higher,with the range from 51.1%to 55.2%,while trnH-psbAhad the lowest GC content（32.0%）.Above all six DNA barcode markers were blasted in NCBI（National Center for Biotechnology Information）,the results showed that the sequences similarity was more than 99%between the wild species of Actinidia and the species of Actinidialogged on NCBI,and there were different similarity levels among the different DNA barcode.Thus,these results revealed the intraspecific diversity of Actinidiaor penetration among different species of Actinidia.Then,the ITSmarker could be used to identify all the above 5species of Actinidiaaccording to the NJ（Neighbor-Joining）tree.And the evolutionary divergence among 5species was obvious in ITSsequences.The trnL-F,ITSand matKbarcode had higher diversity of nucleotide and Tajima＇s test value.By the means of sequence analysis with the six DNA markers,ITSpresented excellently in variation of Actinidia,because of its higher nucleotide diversity and obvious difference among different taxa.

关 键 词：猕猴桃 野生种 DNA条形码 筛选 

分 类 号：S663.4[农业科学—果树学]