基于查尔酮合成酶基因序列多态性鉴定红花不同种质资源

Identification of Different Carthamus tinctorius Germplasm Resources Based on Gene Polymorphism of Chalcone Synthase Gene

目的:开展查尔酮合成酶基因序列多态性研究,以便更准确地对红花种内不同种质资源进行分子鉴定。方法:研究选用全国6个省收集到29份红花种质资源,对常用条形码片段(ITS2、psbA、ITS、ycf5及rpoC1)及查尔酮合成酶基因片段(CtCHS1)进行克隆,采用DnaSP 4.1分析序列多态性,采用软件MAGA 5.0判断克隆片段对红花不同种质资源进行鉴定。结果:psbA、ITS及ycf5对29份红花种质资源的克隆序列完全一致,不能区分29份材料;ITS2序列存在1个位点差异,将29份材料分为2类;rpoC1序列存在4个位点差异,将29份材料分为4类;CtCHS1序列存在23个位点差异,能将29份材料分为13类。结论:研究结果表明基于查尔酮合成酶基因序列多态性能对红花不同种质资源进行鉴定,结果丰富了中药条形码鉴定系统,为后续开发功能标记应用于红花分子辅助选育奠定了基础。

Objective:To analyze chalcone synthase gene sequence polymorphism,in order to accurately identify the different germplasm resources of Carthamus tinctorius.Methods:29 Carthamus tinctorius cultivars collected from six provinces of China were used,and the barcode fragment(ITS2,psbA,ITS,ycf5 and rpoC1)and chalcone synthase gene fragment(CtCHS1)were cloned.DnaSP 4.1 was used to analyze the sequence polymorphism.MAGA 5.0 was used to determine the ability of identification in different germplasm resources.Results:The results showed that the sequences of psbA,ITS and ycf5 were identical and could not be used to discriminate the 29 Carthamus tinctorius cultivars.There was one locus difference in the ITS2,which could divide the 29 Carthamus tinctorius cultivars into two groups.There were four loci differences in the rpoC1 sequence,29 Carthamus tinctorius cultivars could be divided into four groups.In CtCHS1 sequence,there were 23 loci differences,29 Carthamus tinctorius cultivars could be divided into 13 groups.Conclusion:The results show that the different germplasm resources of Carthamus tinctorius can be identified based on the sequence polymorphism of chalcone synthase gene,which enriched the barcode system in the identification of traditional Chinese medicine and laid the foundation for the development of Carthamus tinctorius molecular breeding.