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微生物 DNA 样本保存与偏远热带泥炭地实地研究中可能出现的假象。

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

机构信息

School of Life Sciences, Arizona State University, Tempe, AZ 85287, United States; Biodesign Institute, Arizona State University, Tempe, AZ 85287, United States.

School of Life Sciences, Arizona State University, Tempe, AZ 85287, United States; Biodesign Institute, Arizona State University, Tempe, AZ 85287, United States.

出版信息

J Microbiol Methods. 2024 Sep;224:106997. doi: 10.1016/j.mimet.2024.106997. Epub 2024 Jul 14.

Abstract

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.

摘要

在宿主和环境研究中调查细菌和古菌微生物群落需要收集和储存样本。许多研究都是在偏远地区进行的,这对这些前提条件提出了挑战。当无法进行冷冻保存时,使用保存缓冲液是一种重要的替代方法,但是对于具有挑战性的化学物质的样本或在暴露于需氧环境(如泥炭样本)时提供快速细菌或古菌生长机会的样本,其有效性需要进行方法评估。在这里,我们结合一种针对泥炭土壤样本的已确定的最佳 DNA 提取试剂盒,测试了几种商业和自制保存缓冲液的应用,并就能够最有效地保存反映原始状态的微生物组的方法提出了建议。在使用非最佳缓冲液或没有缓冲液的情况下,我们观察到在保存后最早三天就开始出现明显的群落转移,降低了多样性和群落均匀度,少数特定门的生物出现了由生长驱动的假象。然而,其他缓冲液相对于原始状态保持了非常接近的组成,我们描述了一些指标来了解它们之间的一些差异。由于保存缓冲液的化学影响,测试其兼容性和可靠性以在不同环境中保存原始细菌和古菌群落至关重要。

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