基于EST-SSR分子标记对乌天麻和红天麻的鉴定
Based on EST-SSR Molecular Marker Designing to Identify G.e lata Bl.f.glauca S.Chow and G.e lata Bl.f.elata
何薇 1胡艺镨 1李胜男 1徐娇 1欧小宏1
作者信息
- 1. 贵州中医药大学中药民族药资源研究院,贵阳 550025
- 折叠
摘要
本研究利用 MISA软件对乌天麻转录组筛选得到 SSR位点并对其特征进行分析,通过设计引物对 25 份乌天麻和红天麻样本进行PCR扩增和聚丙烯酰胺凝胶电泳检测,对扩增条带进行聚类分析.结果表明,利用 MISA 软件共筛选得到EST-SSR位点 12 744 个,发生率 27.9%,其中单核苷酸、二核苷酸、三核苷酸为主要重复类型,复合型和多复合型总数 1 071 个,占比 8.41%.利用Primer 3 软件进行SSR引物设计并随机筛选出 40 对,进行PCR扩增和聚丙烯酰胺凝胶电泳检测,其中 4 对扩增表现出多态性,均为复合型和多复合型 SSR,平均有 3 个等位基因,2.31 个有效等位基因,Shannon'多样性指数为 0.69～1.33,平均为0.87,Nei's多样性指数为 0.499 2～0.678 4,平均为 0.552 8.人工读取扩增条带,利用PCoA分析、聚类树、STRUCTURE分析均能将 25 份天麻样本明显地区分为乌天麻和红天麻.
Abstract
This study used MISA software to screen SSR sites from the transcriptome of G.elata Bl.f.elata,then analyzed the SSR characteristics.Primers for PCR amplification and polyacrylamide gel electrophoresis test,which was used for clustering analysis,with 25 G.elata samples were designed.Results showed 12 744 EST-SSR sites were screened out,with a incidence rate of 27.9%.The main repeat types of EST-SSR were mono-,di-and tri-nucleotide,meanwhile the total number of com-pound and multi-compound repeat types was 1 071,accounting of 8.41%.Primers were designed,40 pairs random primers were used for verification by PCR and polyacrylamide gel electrophoresis,that with the result of 4 pair primers amplification showed polymorphism.These SSRs were all compound or multi-compound types with 3 allele,2.31 effective allele,Shannon'diversity index 0.69-1.33,Nei's diversity index 0.449 2-0.678 4.The amplified bands of manually read,with the analysis of PCoA,cluster and STRUCTURE,could classify the 25 samples into G.elata Bl.f.glauca S.Chow and G.elata Bl.f.elata.
关键词
天麻/分子标记/EST-SSR/鉴定Key words
Gastrodia elata/molecular marker/EST-SSR/identification引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据