鸡粪好氧堆肥过程中细菌群落结构和功能分析

The structure and function analysis of bacterial community during aerobic composting of chicken manure

摘要:

为了明确鸡粪好氧堆肥过程中细菌群落结构和功能的变化,采用高通量测序技术测定了好氧堆肥前、中、后3个时期样品的16S rRNA基因序列,并进行了生物信息学分析。结果表明, 3个堆肥阶段中仅有10%左右的分类操作单元(operational taxonomic units, OTUs)具有阶段特异性;不同发酵阶段细菌α多样性指数ACE、Chao1和Simpson均呈现先升高后降低的趋势,但各阶段间差异不显著(P<0.05)。3个发酵阶段优势菌门类相同,但丰度存在差异。线性判别分析[line discriminant analysis (LDA) effect size, LEfSe]法对细菌生物标志分析表明,从门到属水平共有49个物种,堆肥前期样品组中显著富集的物种最多,后期最少。原核分类群功能注释(functional annotation of prokaryotic taxa,FAPROTAX)对细菌功能多样性分析表明,堆肥前期细菌功能多样性最高,而随着发酵进行细菌功能富集程度增加、多样性降低。该研究为畜禽粪污好氧堆肥过程调控提供理论支撑和技术指导。

关键词:鸡粪;好氧堆肥;细菌群落;微生物功能;高通量测序

Abstract:

In order to determine the changes of bacterial community structure and function in the early, middle and late stage of aerobic composting of chicken manure, high-throughput sequencing and bioinformatics methods were used to determine and analyze the 16S rRNA sequence of samples at different stages of composting. Wayne analysis showed that most of the bacterial OTUs in the three composting stages were the same, and only about 10% of the operational taxonomic units (OTUs) showed stage specificity. The diversity indexes including Ace, Chao1 and Simpson showed a trend of increasing at first, followed by decreasing. However, there was no significant difference among different composting stages (P<0.05). The dominant bacteria groups in three composting stages were analyzed at the phylum and genus levels. The dominant bacteria phyla at three composting stages were the same, but the abundances were different. LEfSe (line discriminant analysis (LDA) effect size) method was used to analyze the bacterial biological markers with statistical differences among three stages of composting. From the phylum to genus level, there were 49 markers with significant differences among different groups. The markers included 12 species, 13 genera, 12 families, 8 orders, 1 boundary, and 1 phylum. The most biomarkers were detected at early stage while the least biomarkers were detected at late stage. The microbial diversity was analyzed at the functional pathway level. The function diversity was the highest in the early stage of composting. Following the composting, the microbial function was enriched relatively while the diversity decreased. This study provides theoretical support and technical guidance for the regulation of livestock manure aerobic composting process.

Key words:chicken manure;aerobic composting;bacterial community;microbial function;high-throughput sequencing