光慈菇药材的特异性PCR鉴定

Specific PCR Identification Between Amana edulis and Its Adulterants

目的:研究光慈菇基原植物老鸦瓣及其混伪品特异性PCR鉴定的可行性。方法:基于ITS序列构建光慈菇及其混伪品的系统发育树,并在对ITS序列分析的基础上,为光慈菇设计了一对特异性PCR引物LYB-CP2s/LYB-CP2a,优选扩增条件,用于区别光慈菇及其混伪品。结果:在系统发育树中光慈菇聚类为一个单系。通过对PCR反应的退火温度、循环次数和DNA模板用量进行优化,并对不同型号的PCR仪和Taq酶进行考察,分别获得光慈菇的特异性PCR反应程序。在PCR产物中,正品出现目的条带,而混淆品不出现条带。结论:特异性PCR鉴别方法稳定性高,能够有效地区别光慈菇与其混伪品。

Objective： To study the feasibility of specific PCR identification between Amana edulis and its common adulterants.Methods： Based on all the ITS sequences,the phylogenetic analyses of Amana edulis and its adulterants were conducted using Bayesian inference and maximum parsimony.And based on the ITS sequences of this species and its adulterants,one pairs of specific primer LYB-CP2s/LYB-CP2a were designed for the PCR identification of Amana edulis,and the PCR amplification conditions including the annealing temperature,cycle numbers,and template DNA dose were optimized to establish specific PCR reaction condition.Results： The phylogenetic analysis of ITS revealed that Amana edulis formed one monophyletic group.The results showed that one DNA fragment was amplified from authenticate species by specific PCR reaction condition,whereas no DNA fragment was amplified from its adulterants under the same reaction condition.Conclusion： Amana edulis can be clearly distinguished from its common adulterants by specific PCR with the above primers.