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为了阐明神农架大九湖湿地的古野生湖北海棠的遗传多样性及其进化关系,本研究采集了14株神农架大九湖湿地的野生湖北海棠和2株宜昌区域的湖北海棠品种,通过核糖体转录间隔区(internal translation spacer,ITS)序列分析技术,对16个样品的基因组DNA进行了ITS-PCR扩增,以14个蔷薇科植物的ITS序列作为对照,进行多重对比和系统进化树分析。序列分析结果表明,16个湖北海棠品种的ITS序列长度介于590~596 bp之间,与Gen Bank已公布的ITS序列高度一致,同源性达到98%以上。系统进化树分析表明,DJH2、DJH5、DJH6、DJH7、DJH10同位于一个大的分支,与丽江山荆子亲缘关系较近;DJH8、DJH12、DJH13则处于一个独立的分支;DJH9、DJH11、DJH15、DJH16同位于另一个大的分支,与毛山荆子和三叶海棠亲缘关系较近;DJH3、DJH4与已登录的湖北海棠标准ITS序列最为接近,亲缘关系最近。本项目研究对于神农架大九湖湿地野生湖北海棠的分类和种质资源保护利用具有一定积极意义。
In order to elucidate the genetic diversity and evolutionary relationship of the ancient wild Begonia in the Daciyuhu Lake wetland in Shennongjia, 14 wild crabapple crabapples and two wild crabapple crabapple varieties from Hubei Province were collected and analyzed by ribosome transcription ITS (internal translation spacer) sequence analysis was used to amplify the genomic DNA of 16 samples. ITS sequences of 14 Rosaceae plants were used as controls, and multiple comparisons and phylogenetic tree analyzes were performed. Sequence analysis showed that ITS sequences of 16 Hubei Begonia cultivars ranged from 590 to 596 bp in length, which was highly consistent with the published ITS sequences of Gen Bank, and the homology was over 98%. Phylogenetic tree analysis showed that DJH2, DJH5, DJH6, DJH7 and DJH10 were located in a large branch, which was closely related to P. yunnanensis; DJH8, DJH12 and DJH13 were in an independent branch; DJH9, DJH11, DJH15, DJH16 is located on the other large branch, which has close genetic relationship with C. angustifolia and Trilobata lobata. DJH3 and DJH4 are closest to the registered ITS sequences of Begonia of Hubei, and have the closest genetic relationship. The research of this project is of certain positive significance for the classification and wild germplasm resources conservation and utilization of the wild Begonia in Hubei Province in the Shijiu Lake wetland of Shennongjia.