川芎内生真菌PCR-DGGE分析条件的建立和优化

Establishment and optimization of PCR-DGGE technique on endophytic fungi for Ligusticum chuanxiong

目的采用PCR-DGGE反应技术体系分析川芎内生真菌的群落结构。方法以栽培的川芎根茎为材料,利用PCR-DGGE技术,对基于ITS区的PCR-DGGE体系的变性剂梯度范围、电泳时间进行了优化。结果最佳变性剂梯度的范围为45%～80%,最佳电泳时间为160 min。结论所用方法简便快捷,为进一步研究川芎内生真菌的群落结构分析奠定了基础。

OBJECTIVE To optimize the ploymerase chain reaction and denature gradient gel electrophoresis（PCR-DGGE） technique that was used to analyze the community composition of endophytic fungi for Ligusticum chuanxiong Hort.METHODS The gradient range and electrophoresis time of PCR-DGGE system based on its region were optimized by using rhizomatic material of cultivated Ligusticum chuanxiong.RESULTS The results suggest that the best denaturant gradient range is from 45% to 80%,the optimal electrophoresis time is 160 min.CONCLUSION This method is simple and quick,and layed a foundation for the further study on the analysis of the community of endophytic fungi for Ligusticum chuanxiong.