摘要
采用转录组测序的方法挖掘防风SSR位点信息并设计特异引物,以期为防风的种质资源遗传多样性研究提供有力依据。对防风种子净度、千粒重、发芽率、活力等种子特性进行统计,并对转录组数据库中长度1 kb以上的12606条unigene,利用MISA软件搜索SSR位点信息,使用Primer 3设计引物,以筛选得到的43对SSR引物分析28份不同来源防风材料的多态性。结果显示不同种源防风在净度、千粒重、发芽率、活力、种子长宽等方面存在差异,其中发芽率、活力的差异较为明显,HB-ZJK1、NMG-CF4、NMG-BT、NMG-HLE1、NMG-CF2的千粒重、发芽率、活力方面明显高于其他种源。在得到的86233条unigene序列中利用MISA软件发现有12606条unigene序列上共包含了15958个SSR位点,SSR位点unigene数占unigene总数的14.62%,平均每5009 bp出现1个SSR位点,SSR位点的重复类型以单核苷酸和二核苷酸为主,G/C、TC/AG是单核苷酸和二核苷酸的优势基元,利用28个不同产地防风材料对所有引物进行筛选,得到条带清晰、稳定多态性好的引物。聚类分析表明,28份防风种质资源的生物性状及遗传多样性的聚类结果大致吻合,相同产地(HB-AG1、HB-AG2、HB-ZJK1、HB-ZJK2)的大多聚在一起,亲缘关系较近。防风转录组中SSR出现频率较高、类型丰富且多态性高,为其种质资源鉴别、遗传图谱构建和分子辅助育种等提供了候选分子标记。
Transcriptome sequencing was employed to mine the simple sequence repeat(SSR)locus information of Saposhnikovia divaricata and design specific primers,which aimed to provide a basis for the research on the genetic diversity of S.divaricata germplasm resources.The seed purity,1000-seed weight,germination rate,and seed vigor were determined.MISA was used to obtain the SSR locus information from 12606 unigene longer than 1 kb in the transcriptome database.Forty-three pairs of SSR primers designed in Primer 3 were used to analyze the polymorphism of 28 S.divaricata samples of different sources.The results showed that there were differences in the seed purity,1000-seed weight,germination rate,vigor,and seed length and width among S.divaricata samples of different sources.Particularly,the germination rate and seed vigor had significant differences,and HB-ZJK1,NMG-CF4,NMG-BT,NMG-HLE1,and NMG-CF2 had significantly higher 1000-seed weight,germination rate,and seed vigor than the samples of other sources.Among the 86233 unigene,12606(14.62%)unigene contained 15958 SSR loci,with one SSR locus every 5009 bp on average.The SSR loci were mainly single nucleotide and dinucleotide repeats,which were dominated by G/C and TC/AG,respectively.All the primers were screened by using 28 S.divaricata sample from different habitats,and the primers corresponding to the amplification products with clear bands and stable polymorphism were obtained.The clustering results of the biological characteristics and genetic diversity of the 28 S.divaricata samples were basically consistent,and the samples of the same origin(HB-AG1,HB-AG2,HB-ZJK1,and HB-ZJK2)generally gathered together and had close genetic relationship.The SSRs in S.divaricata transcriptome has high frequency,rich types,and high polymorphism,which provides candidate molecular markers for the germplasm identification,genetic map construction,and molecular-assisted breeding.
作者
李爽
寇佩雯
许祎珂
刘长乐
孙晓春
李铂
黄文静
LI Shuang;KOU Pei-wen;XU Yi-ke;LIU Chang-le;SUN Xiao-chun;LI Bo;HUANG Wen-jing(State Key Laboratory of Research and Development of Characteristic Resources of Qin Medicine(Cultivation),Shaanxi Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,Shaanxi University of Chinese Medicine,Xianyang 712083,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第3期717-727,共11页
China Journal of Chinese Materia Medica
基金
内蒙古自治区2019年度重大科技专项(2019ZD005)
财政部和农业农村部国家现代农业产业技术体系专项(CARS-21)
2018年中医药公共卫生服务补助专项(财社[2018]43号)
陕西省教育厅青年创新团队建设科研计划项目(21JP031)。
关键词
防风
转录组
SSR标记
种子特性
遗传多样性
Saposhnikovia divaricate
transcriptome
SSR marker
seed characteristics
genetic diversity
