• ISSN 1008-505X
  • CN 11-3996/S
陈霄龙, 郭腾飞, 张倩, 岳克, 张珂珂, 宋晓, 丁世杰, 张水清, 黄绍敏. 稳定同位素核酸探针(DNA-SIP)示踪同化利用玉米秸秆碳的微生物群落[J]. 植物营养与肥料学报. DOI: 10.11674/zwyf.2023413
引用本文: 陈霄龙, 郭腾飞, 张倩, 岳克, 张珂珂, 宋晓, 丁世杰, 张水清, 黄绍敏. 稳定同位素核酸探针(DNA-SIP)示踪同化利用玉米秸秆碳的微生物群落[J]. 植物营养与肥料学报. DOI: 10.11674/zwyf.2023413
CHEN Xiao-long, GUO Teng-fei, ZHANG Qian, YUE Ke, ZHANG Ke-ke, SONG Xiao, DING Shi-jie, ZHANG Shui-qing, HUANG Shao-min. Using DNA stable-isotope probing method tracing the microbial communities assimilating the maize straw-derived carbon[J]. Journal of Plant Nutrition and Fertilizers. DOI: 10.11674/zwyf.2023413
Citation: CHEN Xiao-long, GUO Teng-fei, ZHANG Qian, YUE Ke, ZHANG Ke-ke, SONG Xiao, DING Shi-jie, ZHANG Shui-qing, HUANG Shao-min. Using DNA stable-isotope probing method tracing the microbial communities assimilating the maize straw-derived carbon[J]. Journal of Plant Nutrition and Fertilizers. DOI: 10.11674/zwyf.2023413

稳定同位素核酸探针(DNA-SIP)示踪同化利用玉米秸秆碳的微生物群落

Using DNA stable-isotope probing method tracing the microbial communities assimilating the maize straw-derived carbon

  • 摘要:
    目的 研究秸秆分解过程中同化利用秸秆碳源的微生物生态演替过程,明确参与秸秆分解的主要微生物类群,为秸秆高效利用提供科学依据。
    方法 首先采用13C标记高丰度玉米秸秆。微宇宙室内培养试验供试土壤为华北平原石灰性潮土。共设置三个处理:不添加秸秆(soil)、添加自然丰度秸秆(soil+12C-straw)和添加13C标记秸秆(soil+13C-straw)。培养30天时间,结合同位素示踪与高通量测序等技术,测定秸秆添加对土壤有机碳的激发效应,分析利用秸秆碳源的细菌、真菌群落结构,及其与土壤胞外酶活的关系。
    结果 1)秸秆添加显著提高了土壤CO2排放,添加秸秆对土壤有机碳的激发效应在第1天达到峰值,土壤和秸秆CO2排放比例均在第3天达到峰值。2)整个培养时期共发现参与秸秆碳同化利用的细菌微生物操作分类单元(Operational Taxonomic Units, OTUs) 234个,真菌OTUs24个。细菌OTUs主要属于细菌变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)与绿弯菌门(Chloroflexi),真菌OTUs主要为子囊菌门中的粪壳菌纲(Sordariomycetes)。3)秸秆分解不同时期,微生物群落结构具有显著差异。在整个培养时期中,以热单胞菌属(Thermomonas)与溶杆菌属(Lysobacter)为代表的7个细菌属快速响应秸秆添加,以假黄单胞菌属(Pseudoxanthomonas)与土生单胞菌属(Terrimonas)为代表的6个细菌属延迟响应;两个真菌属枝鼻菌属(Cladorrhinum)与葡萄穗霉属(Stachybotrys)表现为对秸秆添加的快速响应,新赤壳属(Neocosmospora)与unclassified_f_Halosphaeriaceae为延迟响应。4)曼特尔分析表明细菌热单胞菌属(Thermomonas)、溶杆菌属(Lysobacter)、绿弯菌门(Chloroflexi)以及真菌裂壳菌属(Schizothecium)与碳氮转化相关土壤胞外酶活性呈显著正相关关系。
    结论 外源秸秆添加对土壤有机碳产生正激发效应,显著增加了土壤CO2排放。同化秸秆碳源的微生物随着培养时间发生群落演替,细菌的热单胞菌属(Thermomonas)、溶杆菌属(Lysobacter)、绿弯菌门(Chloroflexi)以及真菌的裂壳菌属(Schizothecium)在玉米秸秆分解过程中具有重要作用,可为秸秆高效腐熟菌剂的研发提供依据。

     

    Abstract:
    Objectives This study explored the microbial ecological succession of assimilating straw-derived carbon, identified the microbial taxa involved in the process of straw decomposition to provide a scientific basis for the efficient use of straw resources.
    Methods 13C straw was prepared firstly. A microcosm incubation experiment was performed using calcareous fluvo-aquic soil as the test soil. The three treatments were without straw addition, addition of 12C-straw, and addition of 13C straw in soil, and the incubation period was 30 days. Using 13C-labeled maize straw and high-throughput sequencing, we aimed to investigate The priming effect of straw addition on soil organic carbon, and the bacterial and fungal microbial communities assimilating straw-derived carbon were analyzed by combining utilization of 13C-tracing and high throughput methods, and the soil extracellular enzymes activity were analyzed at the same time.
    Results 1) Straw addition significantly increased soil CO2 flux. The priming effect on soil native organic carbon reached peaks at the first day of incubation, and both the soil and straw-derived CO2 emission proportion reached peaks at day 3. 2) During the 30-day incubation period, a total of 234 bacterial OTUs and 24 fungal OTUs, which utilized straw-derived carbon, were identified. The bacteria OTUs were primarily distributed in the bacterial phyla of Proteobacteria, Actinobacteria, Bacteroidetes and Chloroflexi, and the fungal OTUs were belong to class of Sordariomycetes within Ascomycota. 3) The microbial communities varied significantly along the straw decomposition process. Seven bacterial genera, especially Thermomonas and Lysobacter, respond rapidly to straw addition, and 6 bacterial genera, represented by Pseudoxanthomonas and Terrimonas, were identified as delayed responder. Similarly, fungi Cladorrhinum and Stachybotrys respond rapidly to straw addition, Neocosmospora and unclassified_f_ Halosphaeriaceae were identified as delayed responder. 4) Mantel analysis showed that bacterial Thermomonas, Lysobacter, Chloroflexi and fungal Schizothecium were positively correlated with soil extracellular enzyme activity related to C and N transformation (P<0.05).
    Conclusions Exogenous straw has obvious priming effect on soil organic carbon, and could increase soil CO2 emission significantly. The microbes that assimilate the carbon source of straw showed obvious community succession along the decomposition process, in which bacteria of Thermomonas, Lysobacter, Chloroflexi, and along with fungi of Schizothecium played an important role in the maize straw decomposition.

     

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