ITS、matK 及rbc L 是植物常用 DNA 条形码基因(序列).为筛选适合凌云牛心李的DNA条形码基因,对 9 份凌云牛心李种质进行 ITS、matK 及rbc L 序列 PCR 扩增和序列比对分析.通过序列比对分析,9 份凌云牛心李种质 ITS 序列长度在 713～717 bp 之间,G+C 含量59.33%～60.95%之间,存在 27 个变异位点;matK 序列长度为 935～937 bp 之间,G+C 含量33.33%～33.55%之间,存在 5 个变异位点;rbc L 序列长度为 743 bp,G+C 含量在 42.20%～43.07%之间,存在 3 个变异位点.相对于matK及rbc L序列,ITS序列进化速率较快,存在更为丰富的遗传变异位点,更适宜用来开展凌云牛心李分子鉴定及亲缘关系分析.基于 ITS序列构建了9 份凌云牛心李、22 份其他地区李种质、5 份杏种质和 5 份梅种质的NJ 系统进化树,凌云牛心李与其他李种质亲缘关系较近,其中 4 份凌云牛心李种质与中国宁波的茄皮李及日本的大石早生李聚为一个类群,另 5 份凌云牛心李种质与中国嘉兴的槜李、中国山东的玉皇李、中国贵州的九阡李聚为一个分支.凌云牛心李作为地方优质李品种,在长期种植过程中产生了较丰富的遗传变异.
DNA barcode gene screening and genetic relationship analysis of Lingyun Niuxin plum
ITS,matK,and rbc L are commonly used DNA barcode genes(sequences)in plants.To screen DNA barcode genes suitable for Lingyun Niuxin plum,PCR amplified ITS,matK,and rbc L sequences in 9 Lingyun Niuxin plum germplasms was comparatively analyzed.Through sequence alignment analysis,it was found that the ITS sequence length of 9 Lingyun Niuxin plum germplasms ranged from 713 to 717 bp,with G+C content ranging from 59.33%to 60.95%,with 27 variation sites.The length of the matK sequence was between 935 and 937 bp,with G+C content ranging from 33.33%to 33.55%,with 5 variation sites.The length of the rbcL sequence was 743 bp,with G+C content ranging from 42.20%to 43.07%,with three variation sites.Compared with matK and rbc L sequences,the ITS sequence had a faster evolution rate and more abundant genetic variation,more suitable for molecular identification and genetic relationship analysis of Lingyun Niuxin plum.A NJ phylogenetic tree was constructed based on ITS sequences for 9 Lingyun Niuxin plum germplasm resources,22 plum germplasm resources from other regions,5 apricot germplasm resources,and 5 Vatica mangachapoi germplasm resources.Lingyun Niuxin plum germplasm resources were closely related to other plum germplasm resources,with 4 Lingyun Niuxin plum germplasm resources clustered into a group with Solanaceae plum from Ningbo and Dashi Zaosheng plum from Japan.The other 5 Lingyun Niuxin plum germplasm resources clustered into a branch with Hemp plum from Jiaxing,Yuhuang plum from Shan-dong,and Jiuqian plum from Guizhou,China.Lingyun Niuxin plum,as a high-quality local plum variety,had generated the rich genetic variation during long-term cultivation.
万方数据
维普
