| 长期施用有机和无机肥对黑土nirS型反硝化菌种群结构和丰度的影响 |
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| 引用本文： | 尹昌,范分良,李兆君,宋阿琳,朱平,彭畅,梁永超. 长期施用有机和无机肥对黑土nirS型反硝化菌种群结构和丰度的影响[J]. 环境科学, 2012, 33(11): 3967-3975 |
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| 作者姓名： | 尹昌 范分良 李兆君 宋阿琳 朱平 彭畅 梁永超 |
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| 作者单位： | 1. 中国农业科学院农业资源与农业区划研究所,农业部作物营养与施肥重点实验室,北京100081
2. 吉林省农业科学院资源与环境研究中心,长春,130033 |
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| 基金项目： | 国家自然科学基金项目(41171208,40901119);浙江省重大科技专项(2008C12045-2);国家科技支撑计划项目(2012BAD05B06) |
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| 摘 要： | 利用末端限制性片段长度多态性(T-RFLP)和实时荧光定量PCR(real-time quantitative PCR,Q-PCR)技术,结合反硝化潜势(DEA)和土壤理化性质的测定,探索了长期施用有机和无机肥对公主岭黑土nirS型反硝化细菌的群落结构和丰度的影响.试验设不施肥(CK)、单施有机肥(OM)、单施无机肥(NPK)以及有机肥和无机肥混施(MNPK)等4个处理.结果表明,长期施用有机肥显著增加了土壤的DEA,其中OM、NPK和MNPK处理分别为CK处理的5.92、1.81和6.03倍,而NPK和CK间无差异.有机肥处理增加了黑土nirS型反硝化细菌的丰度,OM、NPK和MNPK处理中nirS基因的拷贝数分别为CK的2.73、1.30和3.98倍;NPK处理对nirS基因的拷贝数影响不显著.T-RFLP图谱显示施用有机肥改变了nirS反硝化细菌的群落结构;相比于非有机肥处理,有机肥处理中增加了一类79 bp的片段类型,显著降低了84 bp的片段类型,并完全抑制了一类99 bp的片段类型,而有机肥处理间和非有机肥处理间的nirS群落结构分别相似.系统发育分析表明:黑土中nirS型反硝化菌主要由α、β和γ-变形菌纲及一些尚未培养的微生物组成,79 bp的片段类型与γ-变形菌纲的假单胞菌科(Pseudomonadaceae)和β-变形菌纲的伯克氏菌目(Burkholderiales)相似,84 bp片段类型与Burkholderiales和红环菌目(Rhodocyclales)相似.相关性分析表明,pH、全磷(TP)、全氮(TN)、总有机碳(TOC)、硝态氮(NO3--N)和铵态氮(NH4+-N)依次与nirS型反硝化细菌的种群丰度(r为0.724～0.922,P<0.05)和DEA(r为0.453～0.938,P<0.01)显著相关,DEA与nirS型反硝化细菌的种群丰度显著线性正相关(r=0.85,P<0.01);冗余度分析表明,除含水量外,TN、TP、pH、TOC、NH4+-N和NO3--N(r为0.440～0.862,P<0.01)依次与nirS型反硝化细菌群落结构的变化显著相关,DEA的变化和nirS型反硝化细菌群落结构的变化亦显著相关(r=0.863,P<0.01).本研究表明相比于无机肥处理,公主岭黑土中nirS型反硝化菌的群落结构与丰度对有机肥处理有更显著的响应,且其群落结构的改变与种群丰度的增加与DEA的提高显著相关.
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| 关 键 词： | 黑土 反硝化菌 nirS 实时荧光定量PCR 冗余分析 末端限制性片段长度多态性 |
| 收稿时间： | 2012-02-16 |
| 修稿时间： | 2012-05-24 |
Influences of Long-term Application of Organic and Inorganic Fertilizers on the Composition and Abundance of nirS-type Denitrifiers in Black Soil |
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| YIN Chang,FAN Fen-liang,LI Zhao-jun,Song A-lin,ZHU Ping,PENG Chang and LIANG Yong-chao. Influences of Long-term Application of Organic and Inorganic Fertilizers on the Composition and Abundance of nirS-type Denitrifiers in Black Soil[J]. Chinese Journal of Environmental Science, 2012, 33(11): 3967-3975 |
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| Authors: | YIN Chang FAN Fen-liang LI Zhao-jun Song A-lin ZHU Ping PENG Chang LIANG Yong-chao |
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| Affiliation: | Key Laboratory of Crop Nutrition and Fertilization, Ministry of Agriculture, Institute of Agricultural Resources and Regional Planning, China Academy of Agricultural Sciences, Beijing 100081, China;Key Laboratory of Crop Nutrition and Fertilization, Ministry of Agriculture, Institute of Agricultural Resources and Regional Planning, China Academy of Agricultural Sciences, Beijing 100081, China;Key Laboratory of Crop Nutrition and Fertilization, Ministry of Agriculture, Institute of Agricultural Resources and Regional Planning, China Academy of Agricultural Sciences, Beijing 100081, China;Key Laboratory of Crop Nutrition and Fertilization, Ministry of Agriculture, Institute of Agricultural Resources and Regional Planning, China Academy of Agricultural Sciences, Beijing 100081, China;Agriculture Environment and Resources Center, Jilin Academy of Agricultural Sciences, Changchun 130033, China;Agriculture Environment and Resources Center, Jilin Academy of Agricultural Sciences, Changchun 130033, China;Key Laboratory of Crop Nutrition and Fertilization, Ministry of Agriculture, Institute of Agricultural Resources and Regional Planning, China Academy of Agricultural Sciences, Beijing 100081, China |
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| Abstract: | |
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| Keywords: | black soil denitrifiers nirS real-time quantitative PCR redundancy analysis T-RFLP |
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