Wolsing Martin, Priemé Anders
Department of Microbiology, Copenhagen University, Sølvgade 83H, Copenhagen K, Denmark.
FEMS Microbiol Ecol. 2004 May 1;48(2):261-71. doi: 10.1016/j.femsec.2004.02.002.
ABSTRACT Temporal and spatial variation of communities of soil denitrifying bacteria at sites receiving mineral fertilizer (60 and 120 kgNha(-1)year(-1)) and cattle manure (75 and 150 kgNha(-1)year(-1)) were explored using terminal restriction fragment length polymorphism (T-RFLP) analyses of PCR amplified nitrite reductase (nirK and nirS) gene fragments. The analyses were done three times during the year: in March, July and October. nirK gene fragments could be amplified in all three months, whereas nirS gene fragments could be amplified only in March. Analysis of similarities in T-RFLP patterns revealed a significant seasonal shift in the community structure of nirK-containing bacteria. Also, sites treated with mineral fertilizer or cattle manure showed different communities of nirK-containing denitrifying bacteria, since the T-RFLP patterns of soils treated with these fertilizers were significantly different. Also, these sites significantly differed from the control plot (no fertilizer treatment), whereas the patterns for low and high N-additions were barely separable from each other. Sequencing and phylogenetic analysis of 54 nirK clones revealed that the major part of the nirK-containing bacteria investigated belonged to a yet uncultivated cluster of denitrifying bacteria.
利用聚合酶链反应（PCR）扩增的亚硝酸还原酶（nirK和nirS）基因片段的末端限制性片段长度多态性（T-RFLP）分析，探究了施用矿物肥料（60和120 kgN·ha⁻¹·年⁻¹）和牛粪（75和150 kgN·ha⁻¹·年⁻¹）的场地土壤反硝化细菌群落的时空变化。全年进行了三次分析：3月、7月和10月。nirK基因片段在这三个月均能扩增，而nirS基因片段仅在3月能扩增。T-RFLP图谱的相似性分析表明，含nirK细菌的群落结构存在显著的季节性变化。此外，施用矿物肥料或牛粪的场地显示出不同的含nirK反硝化细菌群落，因为这些肥料处理土壤的T-RFLP图谱显著不同。而且，这些场地与对照地块（未施肥处理）有显著差异，而低氮和高氮添加量的图谱彼此几乎无法区分。对54个nirK克隆进行测序和系统发育分析表明，所研究的含nirK细菌的主要部分属于一个尚未培养的反硝化细菌簇。
