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东北黑土微生物群落对长期施肥及作物的响应
引用本文:丁建莉,姜 昕,关大伟,马鸣超,赵百锁,周宝库,曹凤明,李 力,李 俊.东北黑土微生物群落对长期施肥及作物的响应[J].中国农业科学,2016,49(22):4408-4418.
作者姓名:丁建莉  姜 昕  关大伟  马鸣超  赵百锁  周宝库  曹凤明  李 力  李 俊
基金项目:国家自然科学基金(41573066)、国家“973”计划(2015CB150506)、国家“863”计划(2013AA102802-04)、农业微生物产品质量安全风险评估专项任务(GJFP2016013)、国家现代农业产业技术体系建设专项(CARS-04)
摘    要:【目的】为表征长达35年轮作与施肥条件下东北黑土微生物群落特征,解析长期施肥及作物对土壤微生物丰度和群落结构的影响,探讨东北黑土微生物群落变化与施肥及不同作物间的相互关系,为进一步改良耕作制度与施肥方式提供依据。【方法】依托黑龙江省农业科学院长期定位试验站,选取玉米和大豆两种作物季的4个不同施肥处理:不施肥处理(CK)、有机肥处理(M)、无机肥处理(NPK)和有机肥配施无机肥处理(MNPK)的耕层土壤为研究对象,处理前加字母m表示玉米季样品,加字母s表示大豆季样品。借助Illumina Miseq高通量测序平台和Real-time PCR技术,以16S rRNA基因为分子标靶,研究作物与施肥方式对黑土中微生物群落结构和丰度的影响,分析群落变化与土壤化学性质的相关性。【结果】玉米季土壤16S rRNA基因拷贝数(6.32×10~8—8.83×10~8/ng DNA)比大豆季的低(0.96×10~9—2.30×10~9/ng DNA);玉米季土壤微生物多样性(ACE指数为3 674.58—4 034.84)也低于大豆季(ACE指数为4 167.47—4 887.36);玉米季的细菌丰度以Acidobacteria(24.47%—27.90%)最高,而大豆季丰度最高的是Proteobacteria(27.78%—34.40%),Bacteroidetes和Actinobacteria丰度在两季作物中差异明显。同一作物季的有机肥无机肥配施处理的16Sr RNA基因拷贝数高于无机肥处理,且有机肥配施无机肥处理的微生物α多样性指数也比无机肥处理的高(sMNPK的Chao1指数比sNPK高出11.89%);不同施肥处理之间群落组成存在差异,Alphaproteobacteria在sMNPK和sNPK处理的相对丰度分别比sCK增加3.31%、5.24%;Gammaproteobacteria在sMNPK和sNPK处理均比sCK处理增加1.72%和1.2%,二者相对丰度变化大。相关性分析显示,16Sr RNA基因拷贝数与土壤硝态氮和速效钾正相关;微生物菌落多样性指数与土壤全氮、硝态氮、铵态氮、有效磷和速效钾等化学性质密切相关。【结论】东北黑土不同作物季和不同施肥均会影响土壤微生物丰度、α多样性和群落结构。有机肥无机肥配施能够有效改变微生物群落结构,提高微生物的丰度和多样性,并提高土壤p H,减缓土壤酸化。

关 键 词:施肥  作物  微生物群落结构  土壤化学性质  高通量测序  黑土
收稿时间:2016-05-13

Responses of Micropopulation in Black Soil of Northeast China to Long-Term Fertilization and Crops
DING Jian-li,JIANG Xin,GUAN Da-wei,MA Ming-chao,ZHAO Bai-suo,ZHOU Bao-ku,CAO Feng-ming,LI Li,LI Jun.Responses of Micropopulation in Black Soil of Northeast China to Long-Term Fertilization and Crops[J].Scientia Agricultura Sinica,2016,49(22):4408-4418.
Authors:DING Jian-li  JIANG Xin  GUAN Da-wei  MA Ming-chao  ZHAO Bai-suo  ZHOU Bao-ku  CAO Feng-ming  LI Li  LI Jun
Affiliation:1.Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081;2.Laboratory of Quality &Safety Risk Assessment for Microbial Products, Ministry of Agriculture, Beijing 100081;3.Institute of    Soil Fertility and Environmental Sources, Heilongjiang Academy of Agricultural Sciences, Harbin 150086
Abstract:【Objective】This experiment was carried out to investigate the characteristics of micropopulation in northeast China black soil under more 35 years of fertilization of two crop seasons. The effects of inorganic fertilizer and manure on the abundance and structure of micropopulation were identified. Black soil micropopulation responses to the interaction of long-term fertilization and crops would be revealed. This study will provide evidences for further enhancing fertilization and tillage method.【Method】Based on a long-term fertilization experiment carried out in Heilongjiang Academy of Agricultural Sciences, four different fertilization treatment samples in soybean and maize growing seasons were selected including samples without fertilizer (CK); manure (M); inorganic fertilizer (NPK); and inorganic fertilizers with manure (MNPK). Letter “m” before treatment represented maize season sample, and letter “s” represented soybean season sample. The Illumina Miseq sequencing and quantitative PCR of 16S rRNA gene were applied to analyze the effects of fertilization and crops on micropopulation in black soil. Correlation analysis was carried out between micropopulation and the soil properties.【Result】The 16S rRNA gene copy numbers in maize growing season (6.32×108-8.83×108/ng DNA) was lower than that in soybean growing season (0.96×109-2.30×109/ng DNA). Alpha diversity in maize growing season (ACE index was between 3 674.58-4 034.84) was lower than that of soybean season (ACE index was between 4 167.47-4 887.36), too. The top phylum was Acidobacteria (24.47%-27.90%) in maize growing season, but it was Proteobacteria (27.78%-34.40%) in soybean growing season. The relative abundances of Bacteroidetes and Actinobacteria were significantly different between two crop growing seasons. The 16S rRNA gene copy numbers in treatment of inorganic fertilizers with manure was greater than that of inorganic fertilizers. Alpha diversity in treatment of inorganic fertilizers with manure was higher than that of inorganic fertilizers (Chao1 index of sMNPK compared with that of sNPK was more 11.89%). The composition of micropopulation in different fertilization treatments of one crop growing season was different. The relative abundances of Alphaproteobacteria in sMNPK and sNPK compared with that of sCK were more 3.31% and 5.24%, Gammaproteobacteria in sMNPK and sNPK were higher 1.72% and 1.20% than that in sCK, and were sensitive to fertilizers. Correlation analysis showed that 16S rRNA gene copy number was positively correlated with soil NO3--N and available K, the diversity index and soil total N, NO3--N, NH4+-N, available P and available K were closely related. 【Conclusion】Results of the research demonstrated that of long-term different fertilizations and different crop growing seasons have effects on microbial richness, α diversity and community structure. Inorganic and organic fertilizers improved the soil pH, slowed down soil acidification, changed microbial structures, increased microbial richness and diversity as well as the metabolic activity of micropopulation.
Keywords:fertilizer  crop  microbial community structure  soil chemical properties  high-throughput sequencing  black soil
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