Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, 361021, China.
University of Chinese Academy of Sciences, Beijing, 100049, China.
Sci China Life Sci. 2018 Dec;61(12):1451-1462. doi: 10.1007/s11427-018-9364-7. Epub 2018 Aug 13.
Microorganisms are major drivers of elemental cycling in the biosphere. Determining the abundance of microbial functional traits involved in the transformation of nutrients, including carbon (C), nitrogen (N), phosphorus (P) and sulfur (S), is critical for assessing microbial functionality in elemental cycling. We developed a high-throughput quantitative-PCR-based chip, Quantitative microbial element cycling (QMEC), for assessing and quantifying the genetic potential of microbiota to mineralize soil organic matter and to release C, N, P and S. QMEC contains 72 primer pairs targeting 64 microbial functional genes for C, N, P, S and methane metabolism. These primer pairs were characterized by high coverage (average of 18-20 phyla covered per gene) and sufficient specificity (>70% match rate) with a relatively low detection limit (7-102 copies per run). QMEC was successfully applied to soil and sediment samples, identifying significantly different structures, abundances and diversities of the functional genes (P<0.05). QMEC was also able to determine absolute gene abundance. QMEC enabled the simultaneous qualitative and quantitative determination of 72 genes from 72 samples in one run, which is promising for comprehensively investigating microbially mediated ecological processes and biogeochemical cycles in various environmental contexts including those of the current global change.
微生物是生物圈中元素循环的主要驱动者。确定参与养分转化的微生物功能特征的丰度,包括碳(C)、氮(N)、磷(P)和硫(S),对于评估元素循环中微生物的功能至关重要。我们开发了一种高通量基于定量聚合酶链式反应(PCR)的芯片,称为量化微生物元素循环(QMEC),用于评估和量化微生物群落将土壤有机质矿化并释放 C、N、P 和 S 的遗传潜力。QMEC 包含 72 对针对 C、N、P、S 和甲烷代谢的微生物功能基因的引物对。这些引物对具有较高的覆盖度(平均每个基因覆盖 18-20 个门)和足够的特异性(>70%的匹配率),检测限相对较低(每个运行 7-102 个拷贝)。QMEC 成功应用于土壤和沉积物样品,确定了功能基因的显著不同结构、丰度和多样性(P<0.05)。QMEC 还能够确定绝对基因丰度。QMEC 能够在一个运行中同时定性和定量测定 72 个样本中的 72 个基因,这对于全面研究各种环境背景下(包括当前全球变化的环境背景)微生物介导的生态过程和生物地球化学循环具有广阔的前景。