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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。
Trichoderma harzianum cultured in a shaker was used as the material, and high-quality genomic DNA was extracted by liquid nitrogen milling and the like, which can be used for AFLP experimental analysis to explore and optimize the enzymatic cleavage-ligation reaction, the pre-amplification and the selective amplification reaction The key factors in the process of establishing the best Trichoderma AFLP test system and reaction conditions. AFLP fingerprinting analysis of strains and fusants stored in different periods using this technology system showed that the fingerprints of the same strain preserved at different times had the same characteristics and the fusion fingerprints had the characteristics of both parental strains. Through the exploration and research on the conditions of Trichoderma AFLP, the AFLP reaction system of Trichoderma genome was established. The amplified polymorphic bands could be completely used for the genetic stability and diversity analysis of Trichoderma.