原油降解微生物的16S rDNA序列研究

16S rDNA sequence study for crude oil degraded by micro-organisms

通过向原油中加入营养源的方法培养出降解菌体系,这些细菌能在21 d内将原油完全乳化降解,用红外光谱仪扫描降解后的原油,结果表明,图谱的4 000～4 500 cm-1和5 500～6 000 cm-1两处原油特征峰消失。采用细菌16S rDNA通用引物和PCR扩增等方法,构建原油降解菌体系16S rDNA克隆文库,从中随机挑选35个克隆子,进行序列测定(约800bp),测序结果进行BLAST比对。结果表明,变形杆菌纲(Proteobacteria)是原油降解体系中的优势微生物种群(65.7%),其中苍白杆菌属(Ochrobactrum sp.)、短波单胞菌属(Brevundimonas sp.)和无色杆菌属(Achromobacter sp.)是重要原油降解菌。因此对16S rDNA克隆文库的分析揭示了宁波港原油降解菌株的组成,为以后利用此类细菌处理原油污染提供了重要依据。

By enriching with the nutrition source,the biodegradation bacterium system was obtained from the crude oil.The system showed the considerable emulsification and biodegradability to the crude oil for 21d,and the Near-infrared Technology was used to scan the characteristic spectra of the crude oil after degradation.The result showed that the characteristic peaks（4 000 cm-1～4 500 cm-1 and 5 500 cm-1～6 000 cm-1）disappeared in near-infrared spectra after degradation.A 16S rDNA clone library from the crude oil biodegradation bacterium system was established by PCR using suitable primers.35 clones（ca.800 bp） was sequenced and blasted.The result showed that the proteobacteria was the predominant group in the system of the crude oil biodegradation bacterium.The sequenced clones of the domain bacteria,Brevundimonas diminnta,Ochrobactrum sp.and Achromobacter sp.were the important biodegradation bacterium for the crude oil.The study on 16S rDNA clone library revealed the composition of biodegradation bacterium for the crude oil and offered an important contribution to the treatment of the crude oil contaminations with these bacteria.