Lou Jun, Yang Li, Wang Haizhen, Wu Laosheng, Xu Jianming
Institute of Soil and Water Resources and Environmental Science, College of Environmental and Resource Sciences, Zhejiang Provincial Key Laboratory of Agricultural Resources and Environment, Zhejiang University, Hangzhou, China.
Department of Environmental Sciences, University of California, Riverside, United States of America.
PeerJ. 2018 Mar 14;6:e4514. doi: 10.7717/peerj.4514. eCollection 2018.
Microbial ecological studies have been remarkably promoted by the high-throughput sequencing approach with explosive information of taxonomy and relative abundance. However, relative abundance does not reflect the quantity of the microbial community and the inter-sample differences among taxa. In this study, we refined and applied an integrated high-throughput absolute abundance quantification (iHAAQ) method to better characterize soil quantitative bacterial community through combining the relative abundance (by high-throughput sequencing) and total bacterial quantities (by quantitative PCR). The proposed iHAAQ method was validated by an internal reference strain EDL933 and a laboratory strain WG5. Application of the iHAAQ method to a soil phenanthrene biodegradation study showed that for some bacterial taxa, the changes of relative and absolute abundances were coincident, while for others the changes were opposite. With the addition of a microbial activity inhibitor (NaN), the absolute abundances of soil bacterial taxa, including several dominant genera of , , and , decreased significantly, but their relative abundances increased after 28 days of incubation. We conclude that the iHAAQ method can offer more comprehensive information to reflect the dynamics of soil bacterial community with both relative and absolute abundances than the relative abundance from high-throughput sequencing alone.
微生物生态学研究因高通量测序方法而得到显著推动，该方法能提供有关分类学和相对丰度的海量信息。然而，相对丰度并不能反映微生物群落的数量以及不同分类群之间的样本间差异。在本研究中，我们改进并应用了一种综合高通量绝对丰度定量（iHAAQ）方法，通过结合（高通量测序得到的）相对丰度和（定量PCR得到的）细菌总量，来更好地表征土壤细菌群落的数量特征。所提出的iHAAQ方法通过内部参考菌株EDL933和实验室菌株WG5进行了验证。将iHAAQ方法应用于一项土壤菲生物降解研究表明，对于某些细菌分类群，相对丰度和绝对丰度的变化是一致的，而对于其他分类群，变化则相反。添加微生物活性抑制剂（NaN）后，包括、和几个优势属在内的土壤细菌分类群的绝对丰度显著下降，但在培养28天后它们的相对丰度增加。我们得出结论，与仅通过高通量测序得到的相对丰度相比，iHAAQ方法能够提供更全面的信息，以反映土壤细菌群落相对丰度和绝对丰度的动态变化。
