基于整合高通量绝对定量法的土壤微生物多样性分析
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1.浙江大学土水资源与环境研究所;2.湖州师范学院生命科学学院

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基金项目:

国家重点研发计划项目(2016YFD0200302)和国家自然科学基金项目(41771344)资助


Analysis of Soil Microbial Community Structure via Integrated High-throughput Absolute Abundance Quantification (iHAAQ) Method
Author:
Affiliation:

1.Institute of Soil and Water Resources and Environmental Science,Zhejiang Provincial Key Laboratory of Agricultural Resources and Environment,Zhejiang University;2.Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Province,College of Life Sciences,Huzhou University,Huzhou

Fund Project:

Supported by the National Key Research and Development Program of China (No. 2016YFD0200302) and the National Natural Science Foundation of China (No. 41771344)

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    摘要:

    目前,常用的高通量测序技术只能得到微生物群落结构的物种种类和相对丰度,而整合高通量绝对定量法(iHAAQ)结合了高通量测序技术和qPCR技术,可以进一步计算得到微生物群落结构的绝对含量。相对丰度和绝对含量均是描述微生物群落结构的必备指标,其中相对丰度适合描述和评价微生物群落结构在单个样本内微生物之间的关系,而绝对含量更适于描述和评价微生物群落结构真实的数量变化以及在样本间微生物之间的关系。本研究基于发表的香蕉土传病害、再造沙地农业生态系统和微生物抑制剂作用3篇文献的高通量测序和qPCR数据,通过整合高通量绝对定量法获得3篇文献中古菌域、细菌域和真核生物域的真菌三类微生物群落结构的绝对含量数据,并进一步计算出这三类微生物整体(简称三域微生物)的绝对含量和相对丰度,从而对土壤微生物群落结构不同物种的相对丰度和绝对含量进行分析,旨在更加深入、准确地揭示土壤微生物群落及其生态功能。结果表明:(1)土壤中细菌的物种丰度和绝对含量高于古菌和真菌,在三域微生物中占据主导地位,缺乏绝对含量指标可能造成对微生物群落结构变化理解的偏差;(2)基于相对丰度和绝对含量数据分析,微生物群落结构α多样性指数结果相同,但β多样性指数结果不同;(3)在香蕉土传病害和再造沙地农业生态系统研究中,三域微生物的PCoA结果与细菌的结果较为相似,表明这两项研究中三域微生物群落结构主要受细菌的影响,但在微生物抑制剂的研究中未发现类似结果。整合高通量绝对定量法可以应用于所有同时进行高通量测序和qPCR测序的研究,在未来的土壤微生物群落结构研究中,采用高通量绝对定量法开展三域微生物群落的物种种类、相对丰度和绝对含量的整体评价,具有重要的生态学意义。

    Abstract:

    【Objective】The high-throughput sequencing technology can easily access the species varieties and the relative abundance of a microbial community. Nevertheless, the relative abundance cannot fully reflect the microbial quantities when using it to evaluate the community. On the other hand, the absolute abundance which is one of the essential parameters for describing a microbial community structure in ecology has been ignored. The absolute abundance can be calculated by integrated high-throughput absolute abundance quantification (iHAAQ); which combines high-throughput sequencing and quantitative PCR (qPCR). Both relative and absolute abundances are essential parameters for describing a microbial community structure in ecology. The relative abundance could describe and evaluate the relationship of specific taxa with others in the same sample, while the absolute abundance as a constant parameter is more suitable for describing and evaluating the quantitative variations of specific taxa in a sample or among samples. Thus, this study aimed to provide a comprehensive and in-depth analysis of soil microbial community through the relative and absolute abundances of the archaeal, bacterial, and eukaryotic (fungi) domains of microorganisms (described as three-domain microorganisms).【Method】Based on data of soil microbial community in the studies of banana panama disease, the reclaimed sandy agricultural ecosystem and microbial inhibitor, the absolute abundances of archaea, bacteria and fungi in three papers were obtained by iHAAQ. Then the absolute and relative abundances of three-domain microorganisms were further calculated. The α- and β-diversity analyses of archaea, bacteria, fungi and three-domain microorganisms were conducted according to their relative and absolute abundances, respectively.【Result】The results showed that: (1) Compared to archaea and fungi, the bacteria dominated the soil microbial community with higher species varieties and absolute abundance. Meanwhile, the lack of absolute abundance parameters might lead to the misunderstanding of the microbial community. (2) The α-diversity indexes calculated by relative and absolute abundances were the same, while the β diversity indexes were different. (3) In the research of banana panama disease and reclaimed sandy agricultural ecosystem, PCoA results of three-domain microorganisms and bacteria were relatively similar, which indicated that the community structures of three-domain microorganisms were mainly affected by bacteria in these two studies. But no similar results were found in the research of microbial inhibitors.【Conclusion】The iHAAQ method can be applied to the studies that perform high-throughput sequencing and qPCR analyses, simultaneously. It is of great ecological significance to study the three-domain microbial community with the species varieties, relative and absolute abundances obtained by the iHAAQ method, and it should be encouraged for future research.

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引用本文

王昌毅,楼 骏,严 康,汪海珍,徐建明.基于整合高通量绝对定量法的土壤微生物多样性分析[J].土壤学报,DOI:10.11766/trxb202010200588,[待发表]
WANG Changyi, LOU Jun, YAN Kang, WANG Haizhen, XU Jianming. Analysis of Soil Microbial Community Structure via Integrated High-throughput Absolute Abundance Quantification (iHAAQ) Method[J]. Acta Pedologica Sinica, DOI:10.11766/trxb202010200588,[In Press]

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  • 收稿日期:2020-10-20
  • 最后修改日期:2021-07-04
  • 录用日期:2021-09-08
  • 在线发布日期: 2021-09-10
  • 出版日期: