基于转录组数据的连翘SSR分子标记开发  

SSR Information Analysis and Molecular Marker Development of Forsythia suspensa based on Transcriptome Sequencing

作　　者：马致静 刘灵娣[1] 温春秀[1] 赵敏 李晓东 姜涛 MA Zhijing;LIU Lingdi;WEN Chunxiu;ZHAO Min;LI Xiaodong;JIANG Tao(Institute of Cash Crops,Hebei Academy of Agriculture and Forestry Science,Shijiazhuang 050051,China;College of Landscape and Ecological Engineering,Hebei University of Engineering,Handan 056038,China)

机构地区：[1]河北省农林科学院经济作物研究所,河北石家庄050051 [2]河北工程大学园林与生态工程学院,河北邯郸056038

出　　处：《特产研究》2025年第2期11-17,共7页Special Wild Economic Animal and Plant Research

基　　金：河北省自然科学基金基础研究重大项目(H2023209084);河北省重大科技支撑计划项目(242N6402Z);财政部和农业农村部:国家现代农业产业技术体系资助(CARS-21);河北省省级科技计划资助(22326301D)。

摘　　要：为开发连翘简单重复序列(Simple sequencerepeats,SSR)分子标记,并应用SSR分子标记进行连翘种质的亲缘关系分析,本研究利用MISA软件分析连翘转录组数据,随机筛选设计SSR引物,利用聚丙烯酰胺凝胶电泳开发连翘SSR分子标记。研究分析显示,共获得217959条Unigene序列,平均序列长度为1169bp,N50达到1754bp;对Unigene序列进行分析,发现了54809个SSR位点,位点发生频率13.44%,平均分布距离8.69 kb;连翘转录组SSR信息中,单碱基重复比例最高(49.18%),其次为双碱基重复(29.67%);连翘转录组中单个SSR位点信息中共包含561种重复基原,A/T(48.61%)、AG/TC(30.61%)分别为优势重复基原类型;序列长度在10~27bp之间的SSR位点占SSR位点总数的96.30%。聚丙烯凝胶电泳筛选获得10对条带清晰、稳定性强且多态性明显的引物。利用10对引物对6份连翘种质进行遗传多样性分析,扩增得到25个等位基因,有效等位基因数平均值为1.86。本研究通过聚丙烯凝胶电泳开发出10对连翘SSR分子标记,能有效区分不同连翘种质资源,同时验证了SSR分子标记是连翘遗传多样性分析的一种高效、稳定的技术手段。In order to develop simple sequence repeats(SSR)molecular markers for Forsythia suspensa,and to analyze the genetic relati-onship of Forsythia germplasm by SSR molecular markers.In this study,MISA software was used to analyze the transcriptome data of For-sythia,SSR primers were randomly screened and designed,and SSR molecular markers of Forsythia were developed by polyacrylamide gel electrophoresis.The results showed that a total of 217959 Unigene sequences were obtained,with an average sequence length of 1169 bp and N50 of 1754 bp.Analysis of Unigenes sequences revealed 54809 SSR loci,with a frequency of 13.44%and an average distribution distance of 8.69 kb.In the SSR of Forsythia transcriptome,the proportion of single base repeats was the highest(49.18%),followed by doub-le base repeats(29.67%).A single SSR locus in the transcriptome of Forsythia contained 561 repeat primitives,A/T(48.61%)and AG/TC(30.61%)were the dominant repeat primitive types.The SSR loci with length between 10 bp and 27 bp accounted for 96.30%of the total SSR loci.Ten pairs of primers with clear bands,strong stability and obvious polymorphism were screened,and 25 alleles were amplified.Ten pairs of primers with clear bands,strong stability and obvious polymorphism were screened.Ten pairs of primers were used to analyze the genetic diversity of six Forsythia germplasms,and 25 alleles were amplified,with an average effective number of alleles of 1.86.By stu-dying the information such as the diversity of amplified genes and the number of effective alleles,10 pairs of primers were determined to be accurate and effective.At the same time,it also shows that SSR markers are an efficient and stable technical means for genetic diversity analysis of Forsythia.

关 键 词：连翘 转录组 SSR分析 分子标记 种质资源 

分 类 号：S685.24[农业科学—观赏园艺]