施肥对设施菜地nirK型反硝化细菌群落结构和丰度的影响  

Effects of different fertilization regimes on abundance and community structure of the nirKtype denitrifying bacteria in greenhouse vegetable soils

作　　者：曾希柏[1] 王亚男[1] 王玉忠[2] 白玲玉[1] 李莲芳[1] 段然[1] 苏世鸣[1] 吴翠霞[1] 

机构地区：[1]中国农业科学院农业环境与可持续发展研究所/农业部农业环境重点实验室,北京100081 [2]甘肃省武威市凉州区农业技术推广中心,甘肃武威733000

出　　处：《应用生态学报》2014年第2期505-514,共10页Chinese Journal of Applied Ecology

基　　金："十二五"国家科技支撑计划项目(2012BAD05B06);中央级公益性科研院所基本科研业务费专项资金资助

摘　　要：采用末端限制性片段多态性分析（T-RFLP）和实时荧光定量PCR（Real-timePCR）方法,研究了甘肃武威设施菜地不同施肥条件下0-20cm、20-40cm土层中土壤nirK型反硝化细菌群落结构和丰度的变化。结果表明：施肥对土壤中nirK型反硝化细菌的群落结构具有明显影响,且对70、156、190bp片段所代表设施菜地土壤优势种群影响最显著。施肥对0-20cm土层nirK型反硝化细菌丰度有明显影响,其最大值出现在全有机肥（M）处理、为每克干土2.16107个拷贝数,分别是对照（CK）和全化肥（NPK）处理的2.04和2.02倍。设施菜地土壤0-20cm与20-40cm土层nirK型反硝化细菌的优势种群及其基因丰度均存在显著差异,且设施菜地土壤中nirK型反硝化细菌的群落结构和丰度与大田差异明显。土壤pH值、有机质及硝酸盐含量均影响nirK型反硝化细菌的群落结构和丰度。系统发育分析结果表明,土壤中除存在与厌氧反硝化细菌亲缘相近的nirK型反硝化微生物外,还存在与好氧反硝化菌亲缘关系相近的nirK型反硝化微生物,如根瘤菌属、苍白杆菌属、土壤杆菌属等。The community structure and abundance of nirKtype denitrifying bacteria in different soil layers （0-20 cm and 20-40 cm） under various fertilization regimes in Wuwei, Gansu Province were investigated by the combination of terminal restriction fragment length polymorphism （T-RFLP） and realtime quantitative PCR. Results showed that the nirKtype denitrifying bacteria community structure was significantly affected by fertilization regimes, especially for 70, 156 and 190 bp T-RFs that represented the dominant populations in greenhouse soil. Fertilization regimes significantly influenced the abundance of nirK gene in the 0-20 cm soil layer with the highest abundance of nirK gene copy number （2.16×107 copies·g-1 soil） detected in the manure treatment （M）, which was 2.04 and 2.02 times of that in the control （CK） and chemical fertilizer （NPK） treatments, respectively. Both the dominant population and abundance of nirKtype denitrifying bacteria in the greenhouse soil were significantly different between the 0-20 cm and 20-40 cm soil layers, and the nirKtype denitrifying bacteria community structure and abundance in the greenhouse soil were obviously different from that in the field. Soil pH, soil organic matter content and nitrateN content had the greatest influence on the bacterial community composition. Phylogenetic analysis indicated that there were not only anaerobic nirKtype denitrifying bacteria in greenhouse soil, but also aerobic denitrifying bacteria, such as Rhizobium, Ochrobactrum, Agrobacterium.

关 键 词：nirK型反硝化细菌 末端限制性片段多态性分析 实时荧光定量PCR 群落结构和丰度 

分 类 号：S626[农业科学—园艺学]