牛樟EST-SSR标记的开发及遗传多态性分析  

Developing EST-SSR Markers in Cinnamomum kanehirae and Analyzing the Genetic Polymorphism

作　　者：郭莺[1] 孟红岩[1] 林文珍[1] 林志楷[1] 刘黎卿[1] GUO Ying;MENG Hongyan;LIN Wenzhen;LIN Zhikai;LIU Liqing(Fujian Institute of Subtropical Botany,Xiamen,Fujian 361006,China)

机构地区：[1]福建省亚热带植物研究所,福建厦门361006

出　　处：《热带作物学报》2018年第8期1561-1569,共9页Chinese Journal of Tropical Crops

基　　金：厦门市对台科技合作项目(No.3502Z20182007;No.3502Z20161234;No.3502Z20131150)

摘　　要：本研究通过测序牛樟叶片cDNA获得17 046条Uni-EST,从中检测出1 316个SSR位点,平均每25.7 kb EST含1个SSR位点。功能注释发现1242条SSR-EST中有690条可被划分为3大功能类、18个功能亚类。牛樟EST-SSR二核苷酸重复类型出现频率最高,三核苷酸次之。184类重复基序中AG/CT出现频率最高(32.4%),其次为AAG/CTT(9.3%)。从1 242条SSR-EST设计SSR引物537对,随机选取150对检测牛樟的多态性及在香樟、大叶樟和沉水樟中的可转移性。结果表明,有44对引物在牛樟中得到特异性扩增,其中17对具有多态性,检测出的等位基因数2~7个,平均3.02个。牛樟SSR标记在香樟、沉水樟和大叶樟的可转移率分别为97.7%、97.7%和95.3%。聚类分析表明,在遗传相似系数为0.76处,14个樟属植物能被划分为5类,聚类结果与植物学分类结果基本吻合。17 046 Uni-ESTs were generated by sequencing cDNA of Cinnamomum kanehirae, of which 1 316 SSR loci were identified with 1 SSR on every 25.7 kb EST. Analysis of GO（gene ontology） showed that 690 SSR-ESTs could be divided into three major functional categories and 18 functional sub-categories. Di-nucleotide repeat was the most abundant repeat type followed by tri-nucleotide repeat in EST of C. kanehirae. In 184 motif types, AG/CT（32.4%） was the most abundant followed by AAG/CTT（9.3%）. Based on the 1 242 SSR-EST, a total of 537 primer pairs were successfully designed. 150 primer pairs were randomly selected to detect the polymorphism in C. kanehirae and test transferability in C. camphora, C. micranthum and C. parthenoxylon. The results showed that 44 primer pairs were specifically amplified in C. kanehirae, of which 17 pairs were polymorphic and the number of alleles detected was 2–7, with an average of 3.02. The transfer rates of C. kanehirae SSR marker in C. camphora, C. micranthum and C. parthenoxylon were 97.7%, 97.7% and 95.3%, respectively. Cluster analysis showed that at the genetic similarity coefficient 0.76, 14 Cinnamomum plant samples could be divided into five groups. The clustering result was in good agreement with the result of botany classification.

关 键 词：牛樟 EST-SSR 标记开发 遗传多态性 

分 类 号：Q949.747.5[生物学—植物学]