长期施用尿素对东北黑土中氨氧化古菌群落的影响  

Effects of Long Term Application of Urea on Ammonia Oxidizing Archaea Community in Black Soil in Northeast China

作　　者：周晶[1,2] 姜昕[1,3] 周宝库[4] 马鸣超[1,3] 关大伟[1,3] 赵百锁 陈三凤[2] 李俊[1,3] 

机构地区：[1]中国农业科学院农业资源与农业区划研究所,北京100081 [2]中国农业大学生物学院,北京100094 [3]农业部微生物产品质量安全风险评估实验室,北京100081 [4]黑龙江省农业科学院土壤肥料与环境资源研究所,哈尔滨150086

出　　处：《中国农业科学》2016年第2期294-304,共11页Scientia Agricultura Sinica

基　　金：国家"973"计划(2015CB150506);国家自然科学基金(41573066;41450004);国家"863"计划(2013AA102802-04);国家现代农业产业技术体系建设专项(CARS-04);农业微生物产品质量安全风险评估专项任务(GJFP2014011)

摘　　要：【目的】表征34年施肥条件下东北黑土中氨氧化古菌(AOA)群落特征,明确不同尿素水平对其丰度和群落结构的影响,并与土壤化学性质进行关联分析,探讨引起影响东北黑土中AOA群落变化的主效环境因子,为进一步揭示黑土土壤硝化作用过程和机制,以及改良施肥方式提供依据。【方法】依托黑龙江省农业科学院34年长期定位试验,选取3组不同施肥处理:无尿素组(对照CK和磷钾处理PK)、一倍尿素组(单施一倍氮肥处理N1,氮磷处理NP,氮磷钾处理NPK和氮钾处理NK)和二倍尿素组(单施二倍氮肥处理N2)共7个处理的耕层土壤为研究对象,借助454高通量测序和Real-time PCR技术,以AOA Arch-amo A基因为分子标靶,解析不同尿素水平对黑土中AOA群落组成和丰度的影响,并对AOA群落结构与环境因子进行相关性分析。【结果】随着尿素施用量的增加,黑土中AOA的Arch-amo A拷贝数由2.64×107/g土壤显著降低至8.34×105/g土壤,且黑土p H是引起其数量降低的直接原因;OTU水平上的聚类和非度量多维度分析(NMDS)结果均表明,相同尿素水平的各处理AOA群落结构相似,不同尿素水平处理间的AOA群落差异显著;冗余分析表明,土壤p H、水溶性有机碳和硝态氮浓度是影响土壤AOA群落变化的主效环境因子(P<0.05);系统发育分析表明,东北黑土中AOA主要有Nitrososphaera和Nitrosotalea两个属:无尿素组中99.3%和一倍尿素组中90.1%的Arch-amo A序列归类于Nitrososphaera,二倍尿素组中67.9%的Arch-amo A序列归类于Nitrosotalea。【结论】本研究表征了长期不同施肥下黑土中的AOA群落特征,通过与土壤化学性质耦合分析,确定了引起黑土中AOA群落变化的主要因素。长期施用氮肥的黑土中,AOA Arch-amo A拷贝数和群落结构受尿素施用量的影响显著,低倍尿素提高其多样性,而高倍尿素则使之降低,土壤p H、有机碳和硝态氮浓度是导致AOA群落差异主要因素。【Objective】 This paper aimed to investigate the characteristics of ammonia oxidizing archaeal（AOA） communities in black soil under long-term fertilization in Northeast China to identify the effects of different doses of urea on the abundance and structure of the AOA community. Combined with the soil physiochemical characteristics, the main driving factors would be revealed. This study provides evidence for further understanding of the nitrification process and mechanism in black soil and enhanced fertilization method.【Method】Based on a 34 year fertilization experiment in Heilongjiang Academy of Agricultural Sciences, three groups of samples were selected with different fertilization including samples without urea（without fertilizer CK and phosphorus-potassium PK）, samples with one-time urea（one time nitrogen N1, nitrogen-phosphorus NP, nitrogen-phosphoruspotassium NPK and nitrogen-potassium NK） and samples with two-time urea（two time nitrogen N2）. We applied 454 pyrosequencing and quantitative PCR of the Arch-amo A gene to analyze the effects of urea on the AOA communities in black soil. A correlation analysis was carried out to reveal the main important factors for determining the AOA community composition. 【Result】It was found that AOA Arch-amo A copy numbers in black soil were obviously reduced from 2.64×107 to 8.34×105 /g soil along with the increase of the urea amount, and p H in black soil was the direct reason for the decrease of the Arch-amo A gene. Both clustering and Nonmetric Multidimensional Scaling（NMDS） analyzed result proved that the AOA community structure in each fertilizer treatment with the same urea inputs was similar with each other, while the AOA community structure in fertilizer treatments with different urea addition was significantly different with each other. And a redundancy analysis indicated that soil p H, concentration of water soluble organic carbon and nitrate were the main environmental factors（P 0.05） affecting the AOA community variation. Wh

关 键 词：尿素施用量 氨氧化古菌 土壤酸化 冗余分析 454测序 

分 类 号：S154.3[农业科学—土壤学] S143.14[农业科学—农业基础科学]