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目的:比较黄草乌及其混淆品滇南草乌ITS序列的差别。方法:分别提取黄草乌及滇南草乌总DNA,进行PCR扩增、扩增产物直接测序,利用DNAStar、ClustalX1.81及MEGA4.0软件进行序列分析。结果:通过对黄草乌及其混淆品的ITS数据矩阵,得到ITS1产生229个一致位点、19个变异位点和13个信息位点;5.8S产生162个一致位点、2个变异位点和1个信息位点;ITS2产生217个一致位点、3个变异位点和1个信息位点。除了5.8S没有碱基的转换和颠换,ITS1存在2个碱基转换和1个颠换,ITS2只存在1个颠换。黄草乌2个居群和滇南草乌3个居群的ITS测序结果经数据矩阵分析后,发现二者ITS2区间第596个碱基处为鉴定二者的稳定的信息位点,黄草乌2个居群的所有样品在此位点的碱基为(C),而滇南草乌3个居群的所有样品在此位点的碱基为(A)。结论:黄草乌与其混淆品滇南草乌的核rDNA的ITS序列存在碱基的差异,有特定的变异位点,且位点变异表现出ITS1大于ITS2,因此,能在分子水平上将二者分开。
OBJECTIVE: To compare the differences of ITS sequences between A. chinensis and its confused species A. chinensis. Methods: The total DNA of Radix Aconitum and Radix Aconiti Preparata was extracted and amplified by PCR. The products were directly sequenced and sequenced using DNAStar, ClustalX1.81 and MEGA4.0 softwares. Results: Based on the ITS data matrix of Huangcaowu and its mixture, 229 consensus sites, 19 variation sites and 13 information sites were generated by ITS1. 5.8S produced 162 consensus sites and 2 variant sites Point and 1 information site; ITS2 produced 217 consensus sites, 3 mutation sites and 1 information site. In addition to 5.8S no base conversion and transversion, there are two base conversion ITS1 and a transversion, ITS2 there is only one transversion. ITS sequencing results of 2 populations of Huangcaowu and 3 populations of A. indicus were analyzed by data matrix and found that the 596th base of the ITS2 interval was a stable information locality for identification of the two populations. All samples of the two populations of Uzbekistan had (C) bases at this site, while all samples of the three populations of A. nipponense had (A) bases at this site. CONCLUSIONS: There is a base variation in the ITS sequence of A. rhamnoides nuclear rDNA ITS with specific variation sites, and the site variation shows that ITS1 is greater than ITS2. Therefore, at the molecular level, Separated.