Real-time PCR quantification of anaerobic ammonia-oxidizing microorganisms in red paddy soil
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摘要: 利用厌氧氨氧化细菌16S rDNA基因特异引物对红壤稻田土壤总DNA 进行扩增、PCR产物纯化、克隆和测序,得到属浮霉菌(Planctomycetes)的厌氧氨氧化细菌的部分16S rDNA序列(长度为502 bp).以含该序列的重组质粒作为荧光定量PCR的标准品,对水稻不同生育期红壤稻田土壤中的厌氧氨氧化细菌数量进行检测.结果表明水稻各生育期内稻田表层和根层土壤中均存在一定数量的厌氧氨氧化细菌,且其数量随水稻生长有所变化.表土中的数量随水稻生长呈逐渐增加的趋势,根层土的数量在水稻生长前期变化不大,孕穗期后明显增加.红壤稻田土壤中存在的厌氧氨氧化细菌对稻田土壤的硝化作用可能具有一定贡献.Abstract: DNA in red paddy soil was amplified by polymerase chain reaction(PCR) through specific primers of 16S rDNA gene of anaerobic ammonia-oxidizer.PCR product was purified,cloned and sequenced.A 502 bp sequence of 16S rDNA gene of anaerobic ammonia-oxidizer belonging to Planctomycetes was obtained.Quantity of anaerobic ammonia-oxidizer in the red paddy soil during the rice growing season was monitored by the real-time PCR.The standard curve was made using the plasmid DNA with this rDNA sequence.The results showed that anaerobic ammonia-oxidizers existed in the surface as well as the root zone of the red paddy soil.The quantity changed during the rice growth season.The quantity of anaerobic ammonia-oxidizers in the surface soil increased gradually during the growing season.On the other hand,in the root zone,the number of the bacteria in the soil increased only at the late stage after booting.The anaerobic ammonia-oxidizers might contribute to the nitrification in the red paddy soil.
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Key words:
- red paddy soil /
- anaerobic ammonium-oxidizer /
- real-time PCR
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[1] DRAAF A A,MULDER A,PETER DE B,et al.Anaerobic oxidation of ammonium is a biologically mediated process[J].Appl Environ Microbiol,1955,61(4):1246-1251. [2] 任宏洋,张代钧,丛丽影,等.一种未见报道的厌氧氨氧化细菌分子生物学鉴定[J].环境工程学报,2008,2 (3):314-318. -
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