一种新厌氧氨氧化菌的16S rRNA基因序列测试

Determination of 16S rRNA Gene Sequence for a New ANAMMOX Bacterial Species

运用序批式试验测定厌氧氨氧化污泥的氨氮和亚硝态氮消耗量,求得厌氧氨氧化活性为9.84×10-4mg.(mg.h)-1,厌氧氨氧化菌消耗NO2--N与NH4+-N之比为1.311;采用分子生物学方法从EGSB反应器颗粒污泥中提取细菌总DNA,经纯化、特异引物PCR扩增、克隆、测序等过程,得到厌氧氨氧化菌部分16S rDNA序列;通过系统发育树分析可以发现,在EGSB反应器中富集得到的厌氧氨氧化菌种(anaerobic ammonium-oxidizingPlanctomycete cquenviron-1)与Candidatus'Anammoxoglobus propionicus'和Candidatus'Jettenia asiatica'同属,anaerobic ammonium-oxidizingPlanctomycete cquenviron-1与其他厌氧氨氧化菌基因序列的同源性最大为93%.结果表明,前期EGSB反应器富集得到了一种新型厌氧氨氧化菌,该菌株命名为anaerobic ammonium-oxidizing Planctomycete cquenviron-1.

The anaerobic ammonium oxidation （ANAMMOX） activity of the sludge was about 9.84 × 10^4 mg· （mg·h）^-1 by measuring the simultaneous consumption of ammonium and nitrite under anoxic conditions in the batch tests. The consumption of NO2^- -N and NH4^＋ -N was 1.311 for ANAMMOX bacteria. The partial 16S rDNA sequence was obtained by using molecule biology methods. Crude DNA of the total bacteria in granular sludge from EGSB reactor was extracted and purified. Then, PCR amplification by using specific primer, clone and sequence determination was performed. ANAMMOX bacterial species（ anaerobic ammonium-oxidizing Planctomycete cquenviron-1 ）which was enrichment cultivated from EGSB reactor were the same genera with Candidatus ＂ Anammoxoglobus propionicus＂ and Candidatus ＂ Jettenia asiatica＂ by analyzing phylogenetic tree. The maximum identities of anaerobic ammonium-oxidizing Planctomycete cquenviron- 1 with other ANAMMOX bacterial species was about 93%. The results showed that a new ANAMMOX bacterial species which was enrichment cultivated from EGSB reactor was found and anaerobic ammonium-oxidizing Planctomycete cquenviron- 1 was denominated.