应用位点特异性扩增反应鉴别垂盆草及其近缘植物

AS-PCR Amplification in Identification of Sedum sarmentosum with Its Relative Species

目的:建立垂盆草药材的DNA分子鉴定方法。方法:不同地区野外采集景天属垂盆草及其近缘药用植物共4种,分别提取总DNA,扩增cp DNA trn L-trn F序列,比对所有样品的该段核苷酸序列,设计特异性鉴别垂盆草及其近缘种的位点特异性引物。结果:垂盆草及其近缘易混淆品的trn L-trn F序列具有多个稳定的SNP位点;选取适宜的变异位点设计的2对特异性引物,在一定的退火温度,只有垂盆草扩增后产生预期长度的阳性扩增产物,而其他3种近缘种在同等条件下不能被有效扩增。结论:所设计的鉴别引物在适宜的反应条件下能特异性鉴别垂盆草及其近缘易混淆植物。

Objective： To develop a scientific and effective method for identification of Sedum sarmentosum and its relative materials easily confused. Methods： DNA templates were extracted from plants,and DNA fragments of cp DNA trn L（tRNA-Leu） gene and trn L-trn F intergenic spacer were amplified and sequenced subsequently. trn L-trn F sequences of all samples were aligned. Results： The allele specific PCR method was studied with the designed allele-specific for distinguishing different samples. The result indicated that300 bp and 500 bp DNA fragments were amplified from Sedum sarmentosum,where no any fragment was amplified from its relative species under the same reaction condition. Conclusion： The primers designed in this study are specific for Sedum sarmentosum from its relative species.