摘要
对何首乌及其常见混淆品的核糖体DNA内转录间隔区（rDNAITS）序列进行了测序分析．结果表明：何首乌与其混淆品毛脉蓼、翼蓼及耳叶牛皮消ITSI区的差异率分别为6．91％、18．10％和42．20％，ITS2区的差异率分别为10．00％、19．40％和43．90％；而何首乌种内各居群间ITS1和ITS2区的差异率分别为0．00％～2．13％和0．00％～1．03％．基于何首乌及其混淆品的rDNAITS序列的差异，找出一个位于ITS2间隔区的何首乌特征性M6I酶切位点，用M6I酶对何首乌rDNAITS序列扩增产物酶切后得到含约531bp和109bp的两片段的聚合酶链反应一限制性片段长度多态性（PCR—RFLP）图谱，而其混淆品的rDNAITS序列扩增产物不能被NsbI酶切，故图谱呈单一条带．利用建立的PCR．RFLP方法可以很好地区分何首乌及其混淆品．
The rDNA internal transcribed spacers （ITS） of Fallopia multiflora （F. multiflora） and its frequent adulterants, such as Fallopia multiflora var. cilliinerve, Pteroxygonum giraldiiwere and Cynanchum auriculatum, were sequenced and analyzed. The results show that in the ITS1 region, the divergences between F. multiflora and the three adulterants are respectively 6.91%, 18.10% and 42. 20%, while in the ITS2 region, the divergences are respectively 10. 00%, 19.40% and 43.90% ; and that the intraspecies ITS1 and ITS2 divergences of F. multi- flora are 0. 00% -2. 13% and 0. 00% -1.03% , respectively. Based on the divergences, a endonuclease Nsb I restriction site specific for F. multiflora was detected in the ITS2 region and was used for PCR-RFLP （Polymerase Chain Reaction and Restriction Fragment-Length Polymorphism） analysis. It is found that the PCR product from F. multiflora rDNA ITS can be cleaved by Nsb I into two fragments of about 531 bp and 109bp, while that from the adulterants cannot be cleaved, and that the PCR-RFLP map still retains the undigested original rDNA ITS pattern. It is thus concluded that the proposed PCR-RFLP method is effective in discriminating F. multiflora from its adulterants.
出处
《华南理工大学学报（自然科学版）》
EI
CAS
CSCD
北大核心
2009年第6期74-78,83,共6页
Journal of South China University of Technology(Natural Science Edition)
基金
广东省自然科学基金资助项目(6104397)
