摘要
【目的】研究不同处理羊粪低温沼气发酵液的细菌群落多样性,为羊粪低温沼气发酵菌剂制备提供理论依据。【方法】对不同处理羊粪沼液总DNA进行16S r DNA V3区PCR扩增,扩增产物进行变性梯度凝胶电泳(DGGE)分离、回收、测序,BLAST软件比对分析,构建发育树;同时利用Biolog-ECO法对沼液样本进行微生物功能多样性分析。【结果】PCR-DGGE方法结果显示,在低温条件下(15℃),在发酵各阶段均存在共有优势菌群,包括梭菌属(Clostridium sp.)、拟杆菌属(Bacteroidales bacterium)、假单胞菌属(Pseudomonas sp.)、Proteiniphilum sp.等,其中菌剂III处理的4号为最优发酵试验组,主要为梭菌属(Clostridium sp.)表现有较高的丰度;Biolog-ECO法分析结果显示,在15℃培养条件下,产气高峰期处理2、3和4中,碳代谢活性相对较高,产气量也比对照组高。通过微生物群落的多样性分析发现,处理2的均匀性指数均高于其它处理组。处理3和4的微生物群落的丰富度、优势度指数相对较高。【结论】在低温条件下(15℃)的不同处理中,添加配伍III(配伍I+2 g ZHC固体菌剂)发酵效果最优。
【Objective】The experiment aims to provide a theoretical basis of low-temperature test in sheep biogas fermentation microbial diversity for the preparation of sheep cryogenic methane fermentation agents.【Method】The total DNA 16 S r DNA sheep manure slurry V3 region by different treatments was amplified by PCR and the amplified products were separated,recycled and sequenced by denaturing gradient gel electrophoresis（ DGGE）,and phylogenetic trees were constructed with BLAST alignment analysis software. At the same time,the microbial functional diversity of biogas slurry samples was analyzed by biolog-eco method.【Result】PCR-DGGE method results showed that at low temperature（ 15℃） in the various stages of fermentation,there were dominant bacteria in each stage of fermentation,including Clostridium sp.,Bacteroidales bacterium,Pseudomonas sp.,and Proteiniphilum sp. Among them,Sample No. 4 was the optimum treatment agents in test group III. Especially clostridium（ Clostridium sp.） showed a higher abundance; Biolog-ECO assay showed that the peak of gas production in 2,3 and 4,the carbon activity was relatively high,and the gas production was higher than the control group at a low temperature（ 15℃）. By microbial community diversity analysis,the process uniformity index of treat 2 was higher then others. The microbial community richness and dominance index of treat 3 and 4 was relatively high.【Conclusion】Compatibility III（ compatibility I ＋ 2 g ZHC solid microbial agent） is the best in different processing at a low temperature（ 15℃）.
出处
《新疆农业科学》
CAS
CSCD
北大核心
2016年第10期1885-1893,共9页
Xinjiang Agricultural Sciences
基金
国家自然科学基金项目(31260016)
国家自然科学基金项目(21666038)~~
