焦磷酸测序鉴定建泽泻方法的建立

Establishment of a Pyrosequencing Method to Identify Alisma Orientalis

目的:建立一种基于内部转录间隔区2(ITS 2)序列突变位点鉴别建泽泻的焦磷酸测序方法。方法:分别提取建泽泻与川泽泻的须根、叶片、叶柄DNA,采用焦磷酸测序方法进行分析鉴定,并通过毛细管电泳测序验证方法的正确性。结果:建立了一种基于ITS 2序列焦磷酸测序鉴定方法,经毛细管电泳测序验证结果准确可靠。建泽泻ITS 2序列中的突变位点A-T的突变频率均不小于83%。结论:焦磷酸测序方法可以准确鉴定不同基源的泽泻物种。

Objective Establish a pyrosequencing method based on the ITS 2 sequence mutation sites to identify Alisma orientalis. Methods DNA was extracted respectively from roots, leaves and petiole of Alisma orientalis and analyzed by pyrosequencing. The cor- rectness of the method was verified by capillary electrophoresis sequencing. Results A pyrosequencing method based on the ITS 2 se- quence was established. This method was verified by capillary electrophoresis sequencing to be accurate and reliable. The mutation fre- quency from A to T site was not less than 83% in the ITS 2 sequence of Alisma Orientalis. Conclusion Pyrosequencing can accurately identify Alisma species from different sources.