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目的:建立何首乌(Fallopia multiflora)DNA分子鉴别技术。方法:对何首乌与其近缘种及其混淆品的trnL-trnF(trnL基因和trnF基因间隔区)序列进行比较分析。结果:何首乌与其近缘种及其混淆品的trnL-trnF差异率为2.1%～22%,何首乌种内各居群间trnL-trnF差异率为0%～1.5%。基于何首乌与其近缘种及其混淆品的trnL-trnF序列的差异,找出一个位于trnL5’-trnL3’间区的何首乌特征性Xba I酶切位点(T↓CTAGA),用Xba I酶对不同采集地的何首乌样品trnL-trnF序列扩增产物酶切后均得到含约804～819 bp和256 bp两个片段的PCR-RFLP图谱,而其混淆品trnL-trnF序列扩增产物因不能被Xba I酶切,图谱呈单一条带。结论:利用建立的PCR-RFLP方法可以很好地区分何首乌及其混淆品植物。
Objective: To establish a DNA molecular identification technique for Fallopia multiflora. Methods: The sequence of trnL-trnF (trnL gene and trnF gene spacer region) of Polygonum multiflorum and its related species and their confused products were comparatively analyzed. Results: The difference of trnL-trnF between Polygonum multiflorum and its related species and their confused products ranged from 2.1% to 22%. The difference of trnL-trnF among the populations of Polygonum multiflorum was 0% -1.5%. Based on the difference of trnL-trnF sequence between Polygonum multiflorum and its related species and their confused products, we found a characteristic Xba I site (T ↓ CTAGA) located in the region between trnL5’-trnL3 ’ PCR-RFLP patterns of two fragments of about 804 ~ 819 bp and 256 bp were obtained after the trnL-trnF sequence amplification products were harvested in different samples of Polygonum multiflorum Thunb., While the product of the trnL-trnF sequence was not Xba I digestion, the map was a single band. Conclusion: Polygonum multiflorum and its mixed plants can be distinguished by using the established PCR-RFLP method.