Shifts储存沉积物中DNA产量和生物群落组成:对古基因组学研究的影响

K. Brasell, X. Pochon, J. Howarth, J. Pearman, A. Zaiko, Lucy Thompson, M. Vandergoes, K. Simon, S. Wood
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引用次数: 2

摘要

湖泊沉积物含有丰富的过去环境信息,这些信息对古湖泊学重建非常有价值。这些研究越来越多地应用现代分子工具靶向沉积DNA (sedDNA)。然而,沉积物岩心采样可能在后勤上很困难,这使得立即对sedDNA进行亚采样具有挑战性。沉积物岩心通常在次采样前被冷藏(4°C)数周或数月。我们研究了湖面沉积物在4°C下储存长达24周后,储存时间对DNA(纯化或作为细胞裂解物)浓度变化和生物群落变化的影响。沉积物样品(约0.22 g,每个时间点一式三份)用纯化的DNA(100或200 ng)或从产生蓝藻毒素结核素的微咸水蓝藻的裂解物或未加钉的样品加标。在24周的时间内,每1-4周对样本进行分析。液滴数字PCR结果显示靶基因结节素合成酶F亚基无明显减少;在24周内,加入纯化DNA的样品的拷贝数减少了一半以上,而加入细胞裂解液的样品的拷贝数减少了一半以上。随着时间的推移,使用元条形码评估的细菌和真核生物群落组成发生了显著变化。在细菌中,蓝藻的信号在5周后变得可以忽略不计,而变形菌的信号则增加了。在真核生物群落中,尾藻在6周后成为优势种。这些数据表明,当沉积物冷藏超过5周时,DNA产量和群落组成数据发生显著变化。这凸显了古基因组学研究对快速亚采样和适当储存沉积物岩心样本的需求。
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Shifts in DNA yield and biological community composition in stored sediment: implications for paleogenomic studies
Lake sediments hold a wealth of information from past environments that is highly valuable for paleolimnological reconstructions. These studies increasingly apply modern molecular tools targeting sedimentary DNA (sedDNA). However, sediment core sampling can be logistically difficult, making immediate subsampling for sedDNA challenging. Sediment cores are often refrigerated (4 °C) for weeks or months before subsampling. We investigated the impact of storage time on changes in DNA (purified or as cell lysate) concentrations and shifts in biological communities following storage of lake surface sediment at 4 °C for up to 24 weeks. Sediment samples (~ 0.22 g, in triplicate per time point) were spiked with purified DNA (100 or 200 ng) or lysate from a brackish water cyanobacterium that produces the cyanotoxin nodularin or non-spiked. Samples were analysed every 1–4 weeks over a 24-week period. Droplet digital PCR showed no significant decrease in the target gene (nodularin synthetase – subunit F; ndaF) over the 24-week period for samples spiked with purified DNA, while copy number decreased by more than half in cell lysate-spiked samples. There was significant change over time in bacteria and eukaryotic community composition assessed using metabarcoding. Amongst bacteria, the cyanobacterial signal became negligible after 5 weeks while Proteobacteria increased. In the eukaryotic community, Cercozoa became dominant after 6 weeks. These data demonstrate that DNA yields and community composition data shift significantly when sediments are stored chilled for more than 5 weeks. This highlights the need for rapid subsampling and appropriate storage of sediment core samples for paleogenomic studies.
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来源期刊
Metabarcoding and Metagenomics
Metabarcoding and Metagenomics Agricultural and Biological Sciences-Animal Science and Zoology
CiteScore
5.40
自引率
0.00%
发文量
25
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