基于不同序列对不同来源浙贝母的鉴定分析

Identification and analysis of Fritillaria thunbergii Miq.based on different sequences

目的:基于ITS2、psbA-trnH、matK序列,探讨不同来源浙贝母的分子鉴定方法及系统进化关系。方法:分别提取7个来源浙贝母的DNA,经PCR扩增及测序,并用MEGA 7.0等软件对测序结果进行分析。结果:ITS2序列未检测到变异位点;psbA-trnH序列共检测到124个变异位点,遗传距离变化范围为0~1.103;matK序列共检测到174个变异位点,遗传距离范围为0~0.289。3个野生种相比,象山野生种与各地栽培种源的遗传距离更小。邻接法(NJ)系统树分为两支,象山野生种和各地栽培种源聚为一类,长兴野生种和临平野生种聚为一类,与遗传距离分析结果一致。结论:基于ITS2、psbA-trnH、matK序列,栽培浙贝母地域分化不明显,有待进一步研究。另外,psbA-trnH和matK序列分析结果表明,3个野生种相比,象山野生种与各地栽培种源的亲缘关系更为接近,提示目前的栽培浙贝母可能均起源于象山,具体演化规律有待后续深入研究。

Objective:ITS2,psbA-trnH and matK were used to investigate molecular methods and evolution relationship of Fritillaria thunbergii Miq.from different sources.Methods:The total DNA was extracted from 7 different sources,and the ITS2,psbA-trnH and matK sequences were amplified by PCR and sequenced.The result was analysed by the software MEGA 7.0.Results:No mutation sites were detected in ITS2 sequence.A total of 124 mutation sites were detected in psbA-trnH sequence,and the genetic distance ranged from 0 to 1.103.And 174 mutation sites were detected in matK sequence,and the distance ranged from 0 to 0.289.Compared with the three wild species,the genetic distance between the wild species of Xiangshan and the cultivated provenances was smaller.The neighbor joining algorithm(NJ) tree was divided into two branches.The wild species of Xiangshan and the cultivated provenances were clustered into one class,and the wild species of Changxing and Linping were clustered into another class,which was consistent with the results of genetic distance analysis.Conclusion:Based on ITS2,psbA-trnH and matK sequences,the regional differentiation of cultivated Fritillaria is not obvious,which needs further study.In addition,the results of psbA-trnH and matK sequence analysis show that the genetic relationship between the wild species of Xiangshan and the cultivated provenances is closer comparing with the three wild species,which suggests that the current cultivated Fritillaria may originate from Xiangshan,but the specific evolution law needs to be further studied.