绍兴黄酒酒曲总DNA的提取及其真菌多样性

王洋怡舟; 李柏林; 欧杰; 赵勇; 刘海泉

doi:10.13386/j.issn1002-0306.2016.15.026

绍兴黄酒酒曲总DNA的提取及其真菌多样性

Extracting total DNA in Shaoxing rice wine wheat starter and its fungal diversity

摘要

摘要: 目的:对于黄酒酒曲样品,选择一种合适的总DNA提取方法,并对其中真菌群落结构进行多样性分析。方法:因其质地,结合以往DNA提取方法,比较分析不同方法所获得的总DNA纯度、得率及凝胶电泳图,对实验结果较好的DNA样本进行真菌引物PCR扩增后纯化定量,再经Miseq高通量测序后进行生物信息分析。结果:实验结果表明,使用试剂盒+物理化学酶法获得了较高质量的样品总DNA,A₂₆₀/A₂₈₀比值也接近理想水平,PCR扩增后凝胶电泳图能够得到目的条带,OTU分类、Shannon曲线和系统进化树图显示了其真菌多样性。结论:通过数据的比较和分析,确立了选择的方法是一种较理想的酒曲总DNA的提取方法,对其真菌群落的测定及分析,也为后期开拓黄酒微生物资源提供了有力工具。

Abstract: Objective: To choose appropriate extraction method of total DNA of yellow rice wine and analyze its fugal diversity.Methods: Because of its character,different methods detection were compared and analyzed in this study,including total DNA purity,yield and gel electrophoresis combining with previous DNA extraction method.Objective bands were purified and quantified after PCR amplification of fungi primer,and conducted biological information analysis after Miseq high throughput sequencing.Results: Experimental result showed that kit + Physical+ enzyme method could get better quality and concentration fungal DNA,A_(260)/ A_(280) ratio was close to the ideal level,and could get obvious target band figure after PCR amplification gel electrophoresis. OTU classification,Shannon diversity curve and phylogenetic tree graph showed fungal diversity.Conclusion: Through the comparison and analysis of data,established method in this paper was an ideal method to extract total DNA in the rice wine wheat starter,determination of the fungal community also provided a powerful tool for later developing rice wine microbial resources.