不同悬钩子属植物ITS序列的多态性分析  

Polymorphism Analysis of ITS Sequences in Different Rubus Genera

作　　者：高柱 郭芙蓉 杨静远 汪琪 GAO Zhu;GUO Fu-rong;YANG Jing-yuan(Anhui Guoke Testing Technology Co.,Ltd.,Hefei,Anhui 230041;School of Life Sciences,Anhui Agricultural University,Hefei,Anhui 230036)

机构地区：[1]安徽国科检测科技有限公司,安徽合肥230041 [2]安徽农业大学生命科学学院,安徽合肥230036

出　　处：《安徽农业科学》2023年第19期95-99,共5页Journal of Anhui Agricultural Sciences

基　　金：安徽省科技重大专项(202103a07020003);安徽省重点研究与开发-标准化专项项目(202104h04020036);安徽省重点研究与开发-面上攻关项目(202104a07020013);安徽省重点研究与开发-生态环境专项项目(202104i07020006)。

摘　　要：以13份悬钩子属植物DNA为模板,用PCR技术克隆获得其ITS序列。测序结果分析表明,ITS序列总长在462~464 bp,其中5.8S、ITS1和ITS2序列长度分别为164、208~210、254~255 bp。序列差异比对分析表明,5.8S序列相对保守,不同悬钩子属植物仅有1个碱基差异;ITS2序列变异较大,GC含量在53.73%~55.12%,存在32个变异位点,其中单核苷酸差异位点14个。基于ITS2序列的系统进化树分析表明,13份悬钩子种质资源可分为4簇,其中簇Ⅰ包含掌叶覆盆子种质资源A9、A29和甜叶覆盆子种质资源GH14,簇Ⅱ为HH2、W5和JF9等山莓种质资源,簇Ⅲ为红树莓及黑莓种质资源,簇Ⅳ为TW1、GA1、S和GB3等插田泡种质资源。ITS2序列聚类分析的结果与形态特征分类的结果是一致的,表明外缘标准的ITS2序列对悬钩子属植物资源进行物种分类和鉴定是可靠的。Thirteen germplasm resources in the genus Rubus were used as materials for internal transcribed spacer(ITS)sequence cloning by PCR methods.Sequence alignment analysis revealed that the full length of ITS sequences ranged from 462 to 464 bp.The sequence lengths of 5.8S,ITS1 and ITS2 were 164 bp,208-210 bp and 254-255 bp,respectively.The comparative analysis of sequence differences showed that the 5.8S sequence was relatively conservative,with only one base difference among different rubus genera.ITS2 sequence was highly variable,with GC content ranging from 53.73%to 55.12%,and there were 32 variation sites,including 14 single nucleotide difference sites.Phylogenetic tree analysis based on ITS2 sequence showed that the germplasm resources collected can be divided into four sub-clusters.Among these germplasm resources,the genetic distance between Rubus chingii Hu A9,A29 and Rubus suavissimus S.Lee GH14 were divided into sub-clusterⅠ.Rubus corchonfolius L.F.HH2,W5 and JF9 were divided into sub-clusterⅡ,and red raspberries and black berries were divided into sub-clusterⅢ.And Rubus coreanus Miq.TW1,GA1,S and GB3 were divided into sub-clusterⅣ.The ITS2 analysis is consistent with the results of morphological feature classification,indicating that the ITS2 sequence is reliable in the classification and identification of species in the genus Rubus.

关 键 词：悬钩子属 ITS序列 克隆 分子鉴定 

分 类 号：S663.2[农业科学—果树学]