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      13C-DNA-SIP Distinguishes the Prokaryotic Community That Metabolizes Soybean Residues Produced Under Different CO 2 Concentrations

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          Abstract

          The amendment of crop residues produced under elevated CO 2 (eCO 2) may alter soil microbial community structure and their functions on residue decomposition and carbon (C) cycling in soil. The key to understanding this process is to elucidate the structure of prokaryotic communities that metabolize crop residues derived from eCO 2. A soil incubation experiment was conducted to explore the response of soil microbial community to the amendment of 13C-labeled soybean residues produced under ambient CO 2 (aCO 2) and eCO 2 . The residues were applied to a Mollisol, followed by 13C-DNA stable isotope probing (SIP) and Illumina sequencing on soil prokaryotic community over time. The structure of residue-metabolizing community differed in response to the amendment of eCO 2- and aCO 2-derived residues after 28 days of incubation. In particular, genera Actinomadura, Nocardia, Non-omuraea, and Shimazuella were the dominant members of the residue-metabolizing bacteria, which contributed to this difference. The relative abundances of genera Actinomadura, Nocardia and Shimazuella were 118–144%, 71–113%, and 2–4-fold higher in the Mollisol amended with aCO 2-derived than eCO 2-derived residue. In contrast, the relative abundance of Non-omuraea was 87–90% greater in the eCO 2-residue treatment. However, during the incubation period, there was no difference between the two residue treatments in the community structure as a whole without SIP. These results implied that a pioneering prokaryotic community metabolized the residue initially prior to the entire community. Those bacteria genera being inhibited with the amendment of the eCO 2-derived residue, compared to aCO 2-derived residue, were likely preferential to metabolize recalcitrant C, which might be associated with changes of chemical composition of the residue under eCO 2.

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          Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample.

          The ongoing revolution in high-throughput sequencing continues to democratize the ability of small groups of investigators to map the microbial component of the biosphere. In particular, the coevolution of new sequencing platforms and new software tools allows data acquisition and analysis on an unprecedented scale. Here we report the next stage in this coevolutionary arms race, using the Illumina GAIIx platform to sequence a diverse array of 25 environmental samples and three known "mock communities" at a depth averaging 3.1 million reads per sample. We demonstrate excellent consistency in taxonomic recovery and recapture diversity patterns that were previously reported on the basis of metaanalysis of many studies from the literature (notably, the saline/nonsaline split in environmental samples and the split between host-associated and free-living communities). We also demonstrate that 2,000 Illumina single-end reads are sufficient to recapture the same relationships among samples that we observe with the full dataset. The results thus open up the possibility of conducting large-scale studies analyzing thousands of samples simultaneously to survey microbial communities at an unprecedented spatial and temporal resolution.
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            Stable-isotope probing as a tool in microbial ecology.

            Microorganisms are responsible for driving the biogeochemical cycling of elements on Earth. Despite their importance and vast diversity, the taxonomic identity of the microorganisms involved in any specific process has usually been confined to that small fraction of the microbiota that has been isolated and cultivated. The recent coupling of molecular biological methods with stable-isotope abundance in biomarkers has provided a cultivation-independent means of linking the identity of bacteria with their function in the environment. Here we show that 13C-DNA, produced during the growth of metabolically distinct microbial groups on a 13C-enriched carbon source, can be resolved from 12C-DNA by density-gradient centrifugation. DNA isolated from the target group of microorganisms can be characterized taxonomically and functionally by gene probing and sequence analysis. Application of this technique to investigate methanol-utilizing microorganisms in soil demonstrated the involvement of members of two phylogenetically distinct groups of eubacteria; the alpha-proteobacterial and Acidobacterium lineages. Stable-isotope probing thus offers a powerful new technique for identifying microorganisms that are actively involved in specific metabolic processes under conditions which approach those occurring in situ.
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              Functional responses of plants to elevated atmospheric CO2- do photosynthetic and productivity data from FACE experiments support early predictions?

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                Author and article information

                Contributors
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                24 September 2019
                2019
                : 10
                : 2184
                Affiliations
                [1] 1Key Laboratory of Mollisols Agroecology, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences , Harbin, China
                [2] 2Centre for Experiment, Guizhou University of Traditional Chinese Medicine , Guiyang, China
                [3] 3Centre for AgriBioscience, La Trobe University , Bundoora, VIC, Australia
                Author notes

                Edited by: Malin Bomberg, VTT Technical Research Centre of Finland Ltd., Finland

                Reviewed by: Xuesong Luo, Huazhong Agricultural University, China; Lauren Hale, Agricultural Research Service, United States Department of Agriculture, United States

                *Correspondence: Jian Jin, jinjian29@ 123456hotmail.com

                This article was submitted to Terrestrial Microbiology, a section of the journal Frontiers in Microbiology

                Article
                10.3389/fmicb.2019.02184
                6798031
                cb610c27-2def-4b7e-ac94-540e656f757c
                Copyright © 2019 Wang, Yu, Li, Wang, Tang, Liu, Liu, Xie and Jin.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 18 April 2019
                : 05 September 2019
                Page count
                Figures: 3, Tables: 5, Equations: 0, References: 55, Pages: 10, Words: 0
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                black soil,carbon cycle,crop residue,isopycnic centrifugation,miseq sequencing

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