膜荚黄芪和蒙古黄芪的SSR鉴定

Identification of Astragalus membranaceus and Astragalus membranaceus var.mongholicus Based on SSR Marker

目的:分析蒙古黄芪转录组序列中的SSR位点,设计引物并对膜荚黄芪和蒙古黄芪进行鉴定。方法:对蒙古黄芪根和叶进行转录组测序、从头组装,利用MISA软件搜索SSR位点并设计引物,随机选取110对引物对蒙古黄芪和膜荚黄芪进行鉴定。结果:蒙古黄芪根、叶转录组共获得74 383条unigene,平均长度781 bp。在长度大于1 kb的18 040条序列中检索到5 640个SSR位点,选取其中110对SSR引物对膜荚黄芪和蒙古黄芪进行鉴定,发现引物CL609在10个膜荚黄芪样品中均能扩增出约700 bp的特异条带,而蒙古黄芪无此条带。结论:蒙古黄芪转录组序列的SSR标记可用于膜荚黄芪和蒙古黄芪的鉴定,这些SSR将为黄芪的遗传分析、种质鉴定、分子标记辅助育种等研究提供有利的工具。

Objective:To analysis the SSR loci information in the transcriptome of Astragalus membranaceus var.mongholicus,and to design primers in order to identify Astragalus membranaceus and Astragalus membranaceus var.mongholicus.Methods:The Illumina HiSeq 2500 sequencing technology was used to obtain transcriptome data of Astragalus membranceus var.mongholicus leaves and roots.SSR loci was searched by using MISA and 110 pairs of primers were randomly selected to discriminate the two species.Results:A total of 74 383 unigenes were obtained.The average length of unigenes was 781 bp.5 640 SSRs were found in 18 040 unigenes which length were more than 1 kb.The SSR markers showed rich in type.Among 110 pairs of primers,a band of 700 bp from primer CL609 was specifically amplified in the 10 tested Astragalus membranaceus and was absent in the 10 tested Astragalus membranaceus var.mongholicus.Conclusion:The SSR markers in the transcriptome of Astragalus membranaceus var.mongholicus can be used to identify Astragalus membranaceus and Astragalus membranaceus var.mongholicus at molecular level,which will benefit to genetic diversity study,germplasm identification and marker-assisted breeding.