浓香型白酒黄水、窖泥中微生物总DNA提取方法比较  

Comparison of total microbial DNA extraction methods in yellow water and pit mud in strong-flavor liquor production

作　　者：潘玲玲[1] 王媚[1] 罗明有 傅小红[2] 徐坤[2] 曹子建[2] 王涛[3] 游玲[3] 冯学愚[2] 邱树毅[1] 

机构地区：[1]贵州大学酿酒与食品工程学院,贵州贵阳550025 [2]成都师范学院化学与生命科学学院,四川温江611130 [3]宜宾学院生命科学与食品工程学院,四川宜宾644000

出　　处：《中国酿造》2016年第4期42-46,共5页China Brewing

基　　金：国家固态酿造工程技术研究中心开放项目(GCKF201115);四川省科技厅支撑计划项目(2013SZ0054);成都师范学院引进人才专项项目(YJRC2012-1);四川省科技应用基础项目(2014JY0117)

摘　　要：为高效地获得浓香型白酒窖池中窖泥和黄水中的微生物总DNA以更好地分析其过程中微生物区系,该文设计了4种不同的总DNA提取方法,分别提取了泸州某知名浓香型白酒企业黄水和窖泥的DNA,并用紫外吸收和PCR-DGGE方法比较了不同提取方法所获得总DNA的纯度及其所代表的原核微生物多样性状况。结果显示,4种提取方法均能从2种样品中提取到纯度较高的DNA;以这些DNA为模板均能扩增出细菌和古菌16S r DNA的部分片段的特异性条带;其DGGE电泳图谱均呈现出较为丰富的条带多样性,但条带存在明显差异。综合评价DNA纯度、扩增效果以及DGGE谱图,对于黄水和窖泥,"酶+SDS+液氮法"提取样品中原核微生物DNA的效果最优。To efficiently obtain total DNA of microrganism from pit mud and yellow water in strong-flavor liquor production and better analyze the microbial flora of the process, four different methods were designed for total DNA extraction. The total DNA was extracted from the yellow water and pit mud of a well-known strong-flavor liquor company at Luzhou with the four methods. UV absorption was used to measure the total DNA puri- ty and PCR-DGGE was used to study the diversity of prokaryotic microorganisms. The results showed that the total DNA extracted from two samples by four kinds of methods all had high purity. Using these DNA as a template, specific bands of 16S rDNA portion fragments of the bacteria and archaeal can be amplified. The DGGE showed abundant diversity of the bands, but the bands were significantly different. In comprehensive consideration from DNA purity, amplification effect and the DGGE spectrum, the method of ＂enzyme＋SDS＋liquid nitrogen＂ was the optimal for two kinds of samples.

关 键 词：浓香型白酒 黄水 窖泥 原核微生物 DNA提取 PCR—DGGE 

分 类 号：TS26[轻工技术与工程—发酵工程]