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成品堆肥和蚯蚓堆肥中的细菌多样性:通过对PCR扩增的16S rRNA基因进行非培养分析揭示的差异

Bacterial diversity in a finished compost and vermicompost: differences revealed by cultivation-independent analyses of PCR-amplified 16S rRNA genes.

作者信息

Fracchia Letizia, Dohrmann Anja B, Martinotti Maria Giovanna, Tebbe Christoph C

机构信息

Department of Chemical, Food, Pharmaceutical and Pharmacological Sciences, University of Eastern Piedmont, Via Bovio 6, 28100, Novara, Italy.

出版信息

Appl Microbiol Biotechnol. 2006 Aug;71(6):942-52. doi: 10.1007/s00253-005-0228-y. Epub 2006 Jan 5.

Abstract

Bacterial communities are important catalysts in the production of composts. Here, it was analysed whether the diversity of bacteria in finished composts is stable and specific for the production process. Single-strand conformation polymorphism (SSCP) based on polymerase chain reaction amplified partial 16S rRNA genes was used to profile and analyse bacterial communities found in total DNA extracted from finished composts. Different batches of compost samples stored over a period of 12 years and a 1-year-old vermicompost were compared to each other. According to digital image analysis, clear differences could be detected between the profiles from compost and vermicompost. Differences between three different periods of compost storage and between replicate vermicompost windrows were only minor. A total of 41 different 16S rRNA genes were identified from the SSCP profiles by DNA sequencing, with the vast majority related to yet-uncultivated bacteria. Sequences retrieved from compost mainly belonged to the phyla Actinobacteria and Firmicutes. In contrast, vermicompost was dominated by bacteria related to uncultured Chloroflexi, Acidobacteria, Bacteroidetes and Gemmatimonadetes. The differences were underscored with specific gene probes and Southern blot hybridizations. The results confirmed that different substrates and composting processes selected for specific bacterial communities in the finished products. The specificity and consistency of the bacterial communities inhabiting the compost materials suggest that cultivation-independent bacterial community analysis is a potentially useful indicator to characterize the quality of finished composts in regard to production processes and effects of storage conditions.

摘要

细菌群落是堆肥生产中的重要催化剂。在此,分析了成品堆肥中细菌的多样性对于生产过程是否稳定且具有特异性。基于聚合酶链反应扩增的部分16S rRNA基因的单链构象多态性(SSCP)被用于对从成品堆肥中提取的总DNA中发现的细菌群落进行分析和剖析。将储存了12年的不同批次堆肥样品与一份1年陈的蚯蚓堆肥进行了相互比较。根据数字图像分析,堆肥和蚯蚓堆肥的图谱之间可检测到明显差异。堆肥储存的三个不同时期之间以及重复的蚯蚓堆肥条垛之间的差异较小。通过DNA测序从SSCP图谱中总共鉴定出41个不同的16S rRNA基因,绝大多数与尚未培养的细菌相关。从堆肥中检索到的序列主要属于放线菌门和厚壁菌门。相比之下,蚯蚓堆肥主要由与未培养的绿弯菌门、酸杆菌门、拟杆菌门和芽单胞菌门相关的细菌主导。通过特异性基因探针和Southern杂交进一步强调了这些差异。结果证实,不同的底物和堆肥过程会选择特定成品中的特定细菌群落。栖息在堆肥材料中的细菌群落的特异性和一致性表明,不依赖培养物的细菌群落分析是一种潜在有用指标,可以表征成品堆肥在生产过程和储存条件影响方面的质量。

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