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目的分析DNA分子鉴定黄芪与其混伪品的方法。方法应用相关试剂和仪器对供试样品进行总DNA提取、引物设计、DNA扩增、测序。结果蒙古黄芪和膜荚黄芪具有基本完全一致的ITS序列(P>0.05),与红芪、紫花苜蓿、蜀葵的ITS序列之间的差异均有统计学意义(P<0.05),黄芪与其5种混伪品的种间序列位点变异个数和信息位点个数分别为112个、98个;蒙古黄芪和膜荚黄芪与其伪混品红芪、紫花苜蓿、蜀葵的种间K-2-P遗传距离为0.001~0.460,其中蒙古黄芪和膜荚黄芪具有最小的种间K-2-P距离,和蓝花棘豆具有相等的遗传距离,和红芪具有稍大的遗传距离,和锦鸡儿具有略大的遗传距离,和紫花苜蓿具有更大一些的遗传距离,和蜀葵具有最大的遗传距离,分别为0.001、0.080、0.153、0.158、0.201、0.460;在和黄芪分支的距离方面,蓝花棘豆比锦鸡儿、红芪、紫花苜蓿、蜀葵更近一些(P<0.05);在分类支上,锦鸡儿和红芪也具有较近的遗传关系(P<0.05)。结论 DNA条形码(ITS2)能够有效鉴定黄芪与其混伪品。
Objective To analyze the method of DNA molecular identification of astragalus and its counterfeit products. Methods Total DNA extraction, primer design, DNA amplification and sequencing were performed on the samples by using relevant reagents and instruments. Results Astragalus membranaceus and Astragalus membranaceus from Mongolia had almost the same sequence of ITS (P> 0.05), and had significant difference with ITS sequences of Radix Hedysari, alfalfa and hollyhock (P <0.05) The number of interspecies loci and the number of information loci were 112 and 98, respectively. The interspecies K-2- (2-methyl-2-pyrimidinyloxy) -induced rates of Astragalus membranaceus, Astragalus membranaceus and its pseudo-compound Radix Hedysari, alfalfa and hollyhock The genetic distance of P was between 0.001 and 0.460, in which the Mongolian Astragalus and Astragalus membranaceus had the smallest distance between species K-2-P and had the same genetic distance with O. japonicus, Had a slightly larger genetic distance, had a greater genetic distance than alfalfa, and Hollyhock had the largest genetic distance, 0.001, 0.080, 0.153, 0.158, 0.201, 0.460, respectively; Ophiopogon japonicus was closer than that of Caragana, Radix Hedysarum, alfalfa and hollyhock (P <0.05). There was a close genetic relationship between Caragana and Radix Hedysari (P <0.05). Conclusion DNA barcode (ITS2) can effectively identify astragalus and its adulterants.