基于细胞水平高通量测序的土壤水分对活性微生物的影响研究  

Effect of soil water status on active microbes at cellular level based on high⁃throughput sequencing

作　　者：马馨怡 高威 李颜 郭晓彬 贾仲君[3] 吴金水[2] 王连峰[1] MA Xinyi;GAO Wei;LI Yan;GUO Xiaobin;JIA Zhongjun;WU Jinshui;WANG Lianfeng(School of Environmental and Chemical Engineering,Dalian Jiaotong University,Dalian 116028,China;Key Laboratory of Agro-ecological Processes in Subtropical Region,Institute of Subtropical Agriculture,Chinese Academy of Sciences,Changsha 410125,China;State Key Laboratory of Soil and Sustainable Agriculture,Institute of Soil Science,Chinese Academy of Sciences,Nanjing 210008,China)

机构地区：[1]大连交通大学环境与化学工程学院,大连116028 [2]中国科学院亚热带农业生态研究所亚热带农业生态过程重点实验室,长沙410125 [3]中国科学院南京土壤研究所土壤与农业可持续发展国家重点实验室,南京210008

出　　处：《生态学报》2024年第16期7160-7171,共12页Acta Ecologica Sinica

基　　金：国家自然科学基金项目(42107366,42130716,U23A2052,41471205)。

摘　　要：细胞水平高通量测序技术是从复杂土壤微生物群落中鉴定活性微生物的有力工具。针对我国东北典型水稻土,采用微宇宙控制试验,设置空白对照、湿润(60%田间持水量)和淹水(100%田间持水量)三种处理,通过土壤微生物细胞提取和高通量测序等技术研究了土壤水分对土壤活性微生物的影响规律。结果表明:土壤水分含量增加显著降低了土壤微生物呼吸活性,淹水土壤CO_(2)累积排放量是湿润土壤的87.1%。高通量测序分析表明,土壤DNA和细胞DNA两种水平均能较好地表征土壤优势的活性微生物类群,湿润和淹水土壤优势的门均为变形菌门(Proteobacteria)和厚壁菌门(Firmicutes),而优势的属主要为假单胞菌属(Pseudomonas)。然而,不同水分状况下土壤活性微生物数量、多样性和群落结构发生明显分异。土壤水分增加显著增加了微生物数量,但降低了活性微生物多样性,优势的活性Pseudomonas在湿润状况下微生物丰度(6.7%)显著低于淹水土壤(11.6%)。微生物共存网络分析表明,高水分状况增加了土壤活性微生物间的负相关关系,进而强化了微生物间的竞争关系。湿润土壤正向网络关联度占比高达86.2%,显著高于淹水土壤(61.4%)。功能预测结果表明,土壤水分对生物固氮和硫相关的微生物代谢过程影响显著,且土壤总微生物水平和活性微生物水平上变化趋势一致。上述结果表明,土壤水分增加通过强化微生物群落间的竞争关系进而影响微生物区系及其功能。因此,适当的减少稻田土壤水分的管理方式对东北水稻土微生物功能稳定性具有重要意义。High⁃throughput sequencing at cellular level is a powerful tool for identifying active microbes in complex soil microbiome.Soil microcosms were constructed with three treatments including CK(control),moisture soil(60%soil water holding capacity)and flooded soil(100%soil water holding capacity)in a typical paddy soil in northeast China.The effects of soil water on active microbes were investigated based on soil microbial cell extraction technology and high⁃throughput sequencing.The findings of this study suggested that the increase in soil moisture content had a significant suppressive effect on soil microbial respiration activity.The cumulative CO2 emissions in flooded soil accounted for 87.1%of that in moisture soil over a 48⁃h incubation period.The analysis of high⁃throughput sequencing further indicated that the predominant active microbiome was accurately identified at both the DNA and cellular DNA levels,which included the key phyla of Proteobacteria and Firmicutes,and key genus of Pseudomonas in CK,moisture and flooded paddy soil.However,the real⁃time qPCR and high⁃throughput sequencing consistently suggested that significant differences in microbial abundance,diversity and community structure were observed under different soil water conditions in paddy soil over a 48⁃h incubation period.The increase in soil moisture content had a significant stimulative effect on soil total microbial abundance,but a suppressive effect on soil active microbial diversity in the incubated paddy soil.The active dominant genus of Pseudomonas in moisture soil condition had a lower relative abundance(6.7%)than that in flooded soil(11.6%).The analysis of the microbial co⁃occurrence network further revealed that an increase in soil moisture content significantly increased the negative correlations between active phylotypes,thereby intensifying their competitive interactions in the paddy soil.The proportion of positive network correlations in moisture soil(86.2%)was higher than that in flooded soil(61.4%)after the in

关 键 词：水稻土 土壤水分 活性微生物 高通量测序 细胞水平 

分 类 号：S152.7[农业科学—土壤学] S154.3[农业科学—农业基础科学]