Bacterial Community Structure During Yard Trimmings Composting
Bacterial Community Structure During Yard Trimmings Composting
复制标题
庭院修剪物堆肥过程中的细菌群落结构
DOI：
10.1007/978-3-662-08724-4_3
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发表时间：
2002
影响因子：
1.7
通讯作者：
C. A. Reddy
中科院分区：
文献类型：
--
作者：
F. C. MichelJr.;T. Marsh;C. A. Reddy
我们小组的长期目标是了解各种堆肥参数如何影响堆肥中的微生物群落结构。在这项研究中，我们使用末端限制性片段长度多态性（T-RFLP）的PCR扩增的16 S rRNA基因分析细菌群落结构在堆肥的庭院修剪。从第0、8、29、64和136天从大型市政设施的堆肥料堆（由树叶、草和灌木按4：2：1的比例组成）中收集的样本中分离出群落DNA。使用靶向细菌结构域16 S rRNA基因的荧光标记引物对DNA进行PCR扩增。产物用HhaI、MspI和Rsal限制性消化以得到细菌群落的指纹图谱。末端限制性片段（TRF）的大小与三个digestion进行了比较，通过计算机模拟扩增和限制性digestion完整的16 S rRNA基因序列的三个片段。T-RFLP模式表明广泛的细菌多样性，在所有的堆肥。观察到的TRFs的一个很大的百分比对应的大小预测的细菌通过计算机模拟消化。将来自三种细菌的片段大小与计算机模拟细菌的预测片段大小进行比较，表明从主要含有革兰氏阴性α、β和γ变形菌的细菌群落（第0天）到含有许多革兰氏阳性芽孢杆菌-梭菌群成员（第8、29和64天）以及CFB和放线菌成员（第29和64天）的群落发生了实质性转变。第8、29和64天鉴定的细菌物种包括之前通过培养从嗜热堆肥中分离的细菌，例如芽孢杆菌和假单胞菌属。以及许多以前在堆肥中没有描述的。丰富的TRFs对应E.大肠杆菌和其他革兰氏阴性γ变形菌，堆肥后的前8天急剧下降。第136天的堆肥含有不同的细菌群，包括与已知的泛菌属和假单胞菌属生物防治剂以及黄单胞菌属和芽孢杆菌属物种一致的许多片段大小。在稳定的第64天和第136天堆肥中观察到细菌的最大多样性，其中分别观察到对应于7个和6个不同系统发育组的成员的115个和111个TRF。
A long-term objective of our group is to understand how various composting parameters affect microbial community structure in composts. In this study, we used terminal restriction fragment length polymorphisms (T-RFLP) of PCR-amplified 16S rRNA genes to analyze bacterial community structure during the composting of yard trimmings. Community DNA was isolated from samples collected on days 0, 8, 29, 64, and 136 from a compost windrow (consisting of leaves, grass, and brush in a 4:2:1 ratio) at a large-scale municipal facility. The DNA was PCR-amplified using fluorescently labeled primers targeted to bacterial domain 16S rRNA genes. The products were restriction-digested with Hhal, Mspl,and Rsal to give fingerprints of the bacterial communities. Terminal restriction fragment (TRF) sizes obtained with the three digestions were compared to the three fragments determined by computer-simulated amplification and restriction digestions of complete 16S rRNA gene sequences. T-RFLP patterns indicated extensive bacterial diversity in all of the composts. A large percentage of the observed TRFs corresponded to sizes predicted for bacteria by computer-simulated digestion. Comparison of fragment sizes from three digestions to those predicted by computer-simulated digestions indicated a substantial shift from a bacterial community containing primarily Gram-negative α, β, and γ Proteobacteria (day 0) to communities containing many members of the Gram-positive Bacillus-Clostridium group (days 8, 29, and 64) and members of the CFB and Actinobacteria (days 29 and 64). Bacterial species identified on days 8, 29, and 64 included those previously isolated from thermophilic composts by cultivation such as Bacillus and Pseudomonas spp. as well as many not previously described in composts. Abundant TRFs corresponding to E. coli and other Gram-negative γ Proteobacteria, decreased dramatically after the first 8 days of composting. The day-136 composts contained a diverse group of bacteria including many fragment sizes consistent with known Pantoea and Pseudomonas biocontrol agents as well as Xanthomonas and Bacillus species. The greatest diversity of bacteria was observed in the stabilized day 64 and 136 composts where 115 and 111 TRFs corresponding to members of 7 and 6 different phylogenetic groups, respectively, were observed.