Tuorto Steven J, Brown Chris M, Bidle Kay D, McGuinness Lora R, Kerkhof Lee J
Department of Marine and Coastal Science, Rutgers University, New Brunswick, New Jersey, United States of America.
Environmental Proteomics N.B. Inc, Sackville, New Brunswick, Canada.
PLoS One. 2015 Dec 28;10(12):e0144686. doi: 10.1371/journal.pone.0144686. eCollection 2015.
This report describes BioDry (patent pending), a method for reliably preserving the biomolecules associated with aquatic microbial biomass samples, without the need of hazardous materials (e.g. liquid nitrogen, preservatives, etc.), freezing, or bulky storage/sampling equipment. Gel electrophoresis analysis of nucleic acid extracts from samples treated in the lab with the BioDry method indicated that molecular integrity was protected in samples stored at room temperature for up to 30 days. Analysis of 16S/18S rRNA genes for presence/absence and relative abundance of microorganisms using both 454-pyrosequencing and TRFLP profiling revealed statistically indistinguishable communities from control samples that were frozen in liquid nitrogen immediately after collection. Seawater and river water biomass samples collected with a portable BioDry "field unit", constructed from off-the-shelf materials and a battery-operated pumping system, also displayed high levels of community rRNA preservation, despite a slight decrease in nucleic acid recovery over the course of storage for 30 days. Functional mRNA and protein pools from the field samples were also effectively conserved with BioDry, as assessed by respective RT-PCR amplification and western blot of ribulose-1-5-bisphosphate carboxylase/oxygenase. Collectively, these results demonstrate that BioDry can adequately preserve a suite of biomolecules from aquatic biomass at ambient temperatures for up to a month, giving it great potential for high resolution sampling in remote locations or on autonomous platforms where space and power are limited.
本报告介绍了BioDry（专利正在申请中），这是一种可靠地保存与水生微生物生物量样本相关的生物分子的方法，无需使用危险材料（如液氮、防腐剂等）、冷冻或笨重的存储/采样设备。对实验室中采用BioDry方法处理的样本的核酸提取物进行凝胶电泳分析表明，在室温下储存长达30天的样本中，分子完整性得到了保护。使用454焦磷酸测序和末端限制性片段长度多态性（TRFLP）分析对微生物的16S/18S rRNA基因的存在/缺失和相对丰度进行分析，结果显示与采集后立即在液氮中冷冻的对照样本相比社区无统计学差异。用现成材料和电池驱动的抽水系统构建的便携式BioDry“现场单元”采集的海水和河水生物量样本，尽管在30天的储存过程中核酸回收率略有下降，但也显示出高水平的群落rRNA保存。通过对核酮糖-1,5-二磷酸羧化酶/加氧酶分别进行RT-PCR扩增和蛋白质印迹分析评估，BioDry也有效地保存了现场样本中的功能性mRNA和蛋白质库。总体而言，这些结果表明，BioDry可以在环境温度下将一系列来自水生生物量的生物分子充分保存长达一个月，这使其在空间和电力有限的偏远地区或自主平台上进行高分辨率采样具有巨大潜力。
