木霉AFLP分子标记技术体系的建立

以摇床培养的木霉菌丝为材料,采用液氮研磨等方法提取到高质量的基因组DNA,能够用于AFLP实验分析,探索和优化酶切-连接反应、预扩增和选择性扩增反应过程中的关键因素,建立了适合木霉AFLP的最佳试验体系和反应条件。利用该技术体系对不同时期保存的菌株及融合子进行AFLP指纹分析,发现不同时期保存的同一菌株指纹相同,融合子指纹兼有亲本菌株的特性。通过对木霉AFLP条件的探索和研究,建立了木霉基因组AFLP反应体系,得到的扩增多态性条带完全能够用于木霉遗传稳定性和多样性分析。

The Establishment of AFLP Markers Technical System for Trichoderma spp.

In order to provide a basis for identification and diversity analysis of Trichoderma spp.through AFLP reaction system,improved method was used to extract high quality genomic DNA with hypha from Trichoderma spp.through liquid nitrogen rubbing,which could be used in amplified fragment length polymorphism(AFLP)analysis,The key factors were explored and optimized among enzyme-digested reaction,ligation,pre-amplification and selective amplification.AFLP fingerprinting of strains stored in different periods and the fusions was analysed with this system.The results showed that the fingerprinting of the same strain in different periods was the same,and the fingerprinting of the fusions had the characters from the parents.Therefore,it was a useful tool to research the identification and genetic diversity in Trichoderma spp.through optimized AFLP system.