偏远热带泥炭地的微生物 DNA 样本保存和可能的实地研究人工制品。

IF 1.7 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS
Mark C. Reynolds , Hinsby Cadillo-Quiroz
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引用次数: 0

摘要

在宿主和环境研究中调查细菌和古生物微生物群落需要收集和储存样本。许多研究都是在遥远的地方进行的,对这些先决条件提出了挑战。在无法进行低温保存的情况下,使用保存缓冲液是一种重要的替代方法,但是,对于化学性质具有挑战性的样品,或者暴露在有氧环境中会导致细菌或古生物快速生长的样品(如泥炭样品),其有效性需要进行方法学评估。在这里,结合已确定的泥炭土样本最佳 DNA 提取试剂盒,我们测试了几种商用缓冲液和一种自制缓冲液的应用情况,并就能最有效地保存反映原始状态的微生物群的方法提出了建议。在使用非最佳缓冲液或不使用缓冲液的处理过程中,我们观察到,早在保存后三天,群落就发生了明显的变化,多样性和群落均匀度降低,少数几个特定门类出现了生长驱动的假象。不过,其他缓冲液的组成与原始状态非常接近,我们描述了几种指标,以了解它们之间的一些差异。由于保存缓冲液的化学效应,测试它们在不同环境中保存原始细菌和古细菌群落的兼容性和可靠性至关重要。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Microbial DNA sample preservation and possible artifacts for field-based research in remote tropical peatlands

Microbial DNA sample preservation and possible artifacts for field-based research in remote tropical peatlands

Surveying bacterial and archaeal microbial communities in host and environmental studies requires the collection and storage of samples. Many studies are conducted in distant locations challenging these prerequisites. The use of preserving buffers is an important alternative when lacking access to cryopreservation, however, its effectivity for samples with challenging chemistry or samples that provide opportunities for fast bacterial or archaeal growth upon exposure to an aerobic environment, like peat samples, requires methodological assessment. Here, in combination with an identified optimal DNA extraction kit for peat soil samples, we test the application of several commercial and a homemade preservation buffer and make recommendations on the method that can most effectively preserve a microbiome reflective of the original state. In treatments with a non-optimal buffer or in the absence, we observed notable community shifts beginning as early as three days post-preservation lowering diversity and community evenness, with growth-driven artifacts from a few specific phyla. However other buffers retain a very close composition relative to the original state, and we described several metrics to understand some variation across them. Due to the chemical effects of preservation buffers, it is critical to test their compatibility and reliability to preserve the original bacterial and archaeal community in different environments.

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来源期刊
Journal of microbiological methods
Journal of microbiological methods 生物-生化研究方法
CiteScore
4.30
自引率
4.50%
发文量
151
审稿时长
29 days
期刊介绍: The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.
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