贝母属植物叶绿体基因组候选SNP位点应用分析

贝母属植物是中国中药材的宝库,由于形态上难以区分,在民间应用、临床应用和中药市场中还存在混淆应用、以次充好的现象,急需开发基于新型分子标记位点的精准检测方法。本研究应用多重序列比对、SNP筛选等生物信息学分析手段,对贝母属15种植物28条叶绿体基因组序列进行分析。结果发现,贝母属叶绿体基因组DNA同源性达97.22%,通过分析共发现SNP位点5879个,其中川贝母类鉴别候选位点71个,川贝母鉴别候选位点4个,瓦布贝母鉴别候选位点37个,太白贝母鉴别候选位点147个,浙贝母鉴别候选位点91个,湖北贝母鉴别候选位点271个,平贝母鉴别候选位点1393个,伊贝母鉴别候选位点89个。本研究还对SNP位点在精准鉴定、精确定量应用方面进行了讨论,可为川贝母中药资源鉴定提供技术支撑。

Candidate SNP Application Analysis of Chloroplast Genome of Fritillaria

Fritillaria is a important source of traditional Chinese medicinal material,but there are also terrible situation of shoddy and confusing application in trading market and clinical application.The precise qualitative and quantitative detection methods based specific and new type of molecular markers,are urgently needed to distinguish species of Fritillaria.In this study,bioinformatics analysis methods such as multiple sequence alignment,SNP screening,are used to analyze 28 chloroplast genome sequences of the 15 botany of Fritillaria L.Results show that the identity of 28 Fritillaria chloroplast genome DNA is 97.22%,and 5879 SNPs were found.We analyzed the species specific SNP further by comparing the allelism,and 71,4,37,147,91,271,1393 and 89’flag-site’were screened out for Fritillariae cirrhosae,F.cirrhosa D.Don,Fritillaria unibracteata var.wabuensis,Fritillaria taipaiensis,Fritillaria thunbergii,Fritillaria hupehensis,and Fritillaria ussuriensis,and Fritillariae pallidiflorae respectively.Moreover,the HRM-based,pyrosequencing-based,and KASP-dPCR-based solution were discussed in SNP detection.This study provided technical support for the identification of traditional Chinese medicine resources.