基于转录组测序的浙江楠EST-SSR分子标记开发  

Development of EST-SSR markers based on transcriptome sequences of Phoebe chekiangensis

作　　者：郑永杰[1] 廖枝锋 刘新亮 张月婷 涂白连 伍艳芳[1] ZHENG Yongjie;LIAO Zhifeng;LIU Xinliang;ZHANG Yueting;TU Bailian;WU Yanfang(Camphor Engineering and Technology Research Center of National Forestry and Grassland Administration,Jiangxi Academy of Forestry,Nanchang Jiangxi 330013,China;Jiangxi Shennong Bencao Technology Co Ltd,Jiujiang Jiangxi 330017,China)

机构地区：[1]江西省林业科学院,国家林业草原樟树工程技术研究中心,江西南昌330013 [2]江西神农本草科技有限公司,江西九江330117

出　　处：《南方林业科学》2024年第5期8-12,共5页South China Forestry Science

基　　金：江西省林业科技创新项目(创新专项[2021]15号);江西省林业科学院青年人才培养项目(2023522701;2023522704)。

摘　　要：浙江楠是我国重要的珍稀濒危植物,开展浙江楠SSR位点分布特征及分子标记开发,可为浙江楠的资源评价和保护利用提供科学依据。研究基于高通量测序技术获得浙江楠转录组数据,利用生物信息学方法分析浙江楠转录组SSR位点,开发浙江楠SSR分子标记。结果表明,在浙江楠转录组中共获得116 582个SSR位点,SSR的发生频率为42.38%,平均距离为1.56 kb。浙江楠SSR位点以单核苷酸(63.42%)和二核苷酸重复(25.58%)为主,核苷酸优势基元优势重复基元排前三位的是A/T、AG/CT、AT/AT;SSR重复基元以10~20次重复为主,占总数的64.3%;SSR序列长度在10~646 bp之间,超过20 bp的SSR位点占SSR总数的29.05%,表明浙江楠转录组SSR位点具有较高的多态性。挑选96对SSR引物进行多态性检测,72对引物能够扩增出条带,扩增成功率为75%,最终开发到35对引物在浙江楠可检测到较高的遗传变异,丰富了楠属现有SSR分子标记。Phoebe chekiangensis is an important rare and endangered plant in China.The distribution of SSR loci and the development of molecular markers in P.chekiangensis provide a scientific basis for the resource evaluation,protection and utilization.The Simple Sequence Repeats(SSRs)loci were developed by bioinformatic analyses of P.chekiangensis transcriptome sequences.A total of 116582 SSR loci were obtained in the transcriptome of P.chekiangensis with the occurrence frequency of 42.38%.The average distribution distance of SSRs was 1.56 kb.The SSR of P.chekiangensis transcriptome was dominated by mononucleotides(63.42%)and dinucleotide repeats(25.58%),and the top three dominant SSR motifs are A/T,AG/CT and AT/AT;SSR repeat motifs are dominated by 10 to 20 repeats,accounting for 64.3%;the length of SSR sequences ranged from 10 to 646 bp,and the SSR loci exceeding 20 bp accounted for 29.05%of the total SSRs,indicating that the SSR loci in the transcriptome of P.chekiangensis have good polymorphism potential.72 of 96 pairs of SSR primers were successfully amplified bands,the amplification achievement ratio was 75%,and 35 pairs of primers showed high polymorphic information content and enriched the existing SSR molecular marker of Phoebe.

关 键 词：浙江楠 SSR重复类型 引物开发 转录组 遗传多样性 

分 类 号：S718.46[农业科学—林学]