Committee on Evolutionary Biology, University of Chicago, Chicago, Illinois, USA.
PLoS One. 2013 Jul 31;8(7):e70460. doi: 10.1371/journal.pone.0070460. Print 2013.
Recent advances in DNA sequencing technologies have allowed scientists to probe increasingly complex biological systems, including the diversity of bacteria in the environment. However, despite a multitude of recent studies incorporating these methods, many questions regarding how environmental samples should be collected and stored still persist. Here, we assess the impact of different soil storage conditions on microbial community composition using Illumina-based 16S rRNA V4 amplicon sequencing. Both storage time and temperature affected bacterial community composition and structure. Frozen samples maintained the highest alpha diversity and differed least in beta diversity, suggesting the utility of cold storage for maintaining consistent communities. Samples stored for intermediate times (three and seven days) had both the highest alpha diversity and the largest differences in overall beta diversity, showing the degree of community change after sample collection. These divergences notwithstanding, differences in neither storage time nor storage temperature substantially altered overall communities relative to more than 500 previously examined soil samples. These results systematically support previous studies and stress the importance of methodological consistency for accurate characterization and comparison of soil microbiological assemblages.
近年来，DNA 测序技术的进步使科学家能够深入研究日益复杂的生物系统，包括环境中的细菌多样性。然而，尽管最近有许多研究采用了这些方法，但关于如何收集和储存环境样本的许多问题仍然存在。在这里，我们使用基于 Illumina 的 16S rRNA V4 扩增子测序来评估不同土壤储存条件对微生物群落组成的影响。储存时间和温度都影响了细菌群落的组成和结构。冷冻样本保持了最高的 alpha 多样性，beta 多样性的差异最小，这表明冷藏对于维持稳定的群落是有用的。储存时间为三天和七天的样本既有最高的 alpha 多样性，又有最大的整体 beta 多样性差异，表明样本采集后群落的变化程度。尽管存在这些差异，但与之前检查的 500 多个土壤样本相比，储存时间和储存温度的差异并没有显著改变总体群落。这些结果系统地支持了以前的研究，并强调了方法一致性对于准确描述和比较土壤微生物组合的重要性。
