High-throughput sequencing shows inconsistent results with a microscope-based analysis of the soil prokaryotic community
High-throughput sequencing shows inconsistent results with a microscope-based analysis of the soil prokaryotic community
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DOI：
10.1016/j.soilbio.2014.05.010
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发表时间：
2014-09
影响因子：
9.7
通讯作者：
Masayuki Ushio;K. Makoto;J. Klaminder;H. Takasu;S. Nakano
中科院分区：
文献类型：
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作者：
Masayuki Ushio;K. Makoto;J. Klaminder;H. Takasu;S. Nakano
在本研究中，我们首次直接分析了原核生物土壤群落的组成如何根据高通量测序或荧光原位杂交(FISH)结合催化报告沉积(CARD)的使用而不同。土壤样本是沿着瑞典北部较短(<；3米)的苔原植被坡度采集的。高通量测序估计的酸性细菌和类杆菌的相对丰度高于CARD-FISH估计的相对丰度，而古生菌和α-Prote杆菌的相对丰度低于CARD-FISH估计的相对丰度。结果表明，如果我们假设卡鱼可以提供潜在的更多定量数据，那么高通量测序高估/低估了一些微生物分类群的相对丰度。在解释分子技术(高通量测序和卡片-FISH)产生的数据时应非常谨慎，需要多种方法的支持才能得出可靠的结论。
In the present study, we perform the first direct analysis on how the composition of the prokaryotic soil community differs depending on whether high-throughput sequencing or fluorescentin situhybridization (FISH) coupled with catalyzed reporter deposition (CARD) is used. Soil samples were collected along short (<3 m) tundra vegetation gradients from Northern Sweden. Relative abundances ofAcidobacteriaandBacteroidetesestimated by the high-throughput sequencing were higher than those estimated by CARD–FISH, while relative abundances ofArchaeaandα-Proteobacteriaestimated by high-throughput sequencing were lower than those estimated by CARD–FISH. The results indicated that the high-throughput sequencing overestimates/underestimates the relative abundance of some microbial taxa if we assume that CARD–FISH can provide potentially more quantitative data. Great caution should be taken when interpreting data generated by molecular technologies (both of high-throughput sequencing and CARD–FISH), and supports by multiple approaches are necessary to make a robust conclusion.