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本文对我国板蓝根及大青叶原植物的物种进行了探讨,以澄清分类上的混乱现象。利用PCR直接测序法对采集的全国22份样本,测定了核基因ITS2区和叶绿体matK基因片段,所得序列经CodonCode Aligner拼接后,用软件MEGA4.0进行相关数据分析,并构建NJ(邻接)树。结果表明,所研究样本ITS2片段长度为191 bp,测序峰图显示不同研究样本间具有多个单核苷酸多态性(SNP)位点,部分样本具有杂合位点;从构建的NJ树图可以看出,基于核ITS2片段,所研究样本被分成两大支,其中一支与欧洲菘蓝相聚。基于叶绿体matK序列,所研究样本也被明显分成两支。因此本研究支持板蓝根及大青叶的原植物为菘蓝(Isatis indigotica Fortune),与欧洲菘蓝(Isatis tinctoria L.)是独立的两个物种,不支持Flora of China中将二者合并的观点。
In this paper, the species of Banlangen and Daqingye in China were discussed in order to clarify the chaos on the classification. PCR-direct sequencing was used to detect the ITS2 region of chloroplast and the matK gene fragment of chloroplast in 22 samples collected from China. The obtained sequences were stitched with CodonCode Aligner and analyzed with software MEGA4.0 to construct NJ (contiguous) . The results showed that the length of the ITS2 fragment was 191 bp. The sequencing peak map showed that there were multiple single nucleotide polymorphisms (SNPs) between different samples and some of the samples had heterozygous loci. From the constructed NJ tree It can be seen from the figure that based on the nuclear ITS2 fragment, the sample studied is divided into two major branches, one of which is clustered with European blue. Based on the chloroplast matK sequence, the sample studied is also clearly divided into two. Therefore, the original plants of Isatis indigotica Fortune and Isatis tinctoria L. supported by this study are two independent species of Isatis tinctoria L. They do not support the idea of combining the two in Flora of China .