摘要
【目的】探究不同浓度石油烃条件下微生物群落结构特征,定向培育高效石油烃降解微生物菌群,并挖掘具有石油烃降解功能的菌株资源。【方法】以0#柴油作为唯一碳源,通过逐级提高其浓度对石油污染土壤样品进行逐级驯化,共进行5代驯化。基于16S rRNA基因扩增子测序技术揭示微生物群落结构变化,利用稀释涂布和平板划线法分离纯化具有石油烃降解潜力的菌株。通过改进2,6-二氯靛酚培养体系验证实验筛选高效降解菌株。【结果】随着驯化过程中0#柴油浓度的升高,在7000 mg/L条件下拟杆菌门(Bacteroidota)、芽孢杆菌门(Bacillota)等具有石油烃降解功能的细菌门相对丰度显著升高。共分离得到58株细菌,分属于4门22科25属。其中,假单胞菌门(Pseudomonadota)31株,占比53.45%;放线菌门(Actinomycetota)13株,占比22.41%;芽孢杆菌门(Bacillota)11株,占比18.97%;拟杆菌门(Bacteroidota)3株,占比5.17%。同时,筛选获得18株具有潜在石油烃降解功能的菌株。【结论】通过逐级驯化培养,成功富集获得7个在7000 mg/L 0#柴油浓度下石油烃降解率超过70%的驯化菌群。扩增子测序表明,不同浓度的0#柴油明显改变了微生物群落结构,并筛选得到18株能以0#柴油作为唯一碳源进行生长的细菌,为微生物修复石油污染土壤提供了潜在的菌种资源。
[Objective]To investigate the structural characteristics of microbial consortia in different concentrations of petroleum hydrocarbons,cultivate efficient petroleum hydrocarbondegrading microbial consortia,and mine the strain resources capable of degrading petroleum hydrocarbons.[Methods]We used 0#diesel as the sole carbon source to domesticate oilcontaminated soil samples through five successive generations by gradually increasing the 0#diesel concentration.The structural changes of microbial consortia were uncovered by 16S rRNA gene amplicon sequencing.The strains with petroleum hydrocarbon-degrading potential were isolated and purified via dilution plating and streaking.Finally,the improved 2,6-dichlorophenol indophenol(DCPIP)cultivation system was employed to identify efficient degrading strains.[Results]During domestication,when the concentration of 0#diesel was raised to 7000 mg/L,the relative abundance of petroleum hydrocarbon-degrading bacteria including Bacteroidota and Bacillota significantly increased.A total of 58 bacterial strains belonging to 25 genera,22 families of 4 phyla were isolated,including 31(53.45%)strains of Pseudomonadota,13(22.41%)strains of Actinomycetota,11(18.97%)strains of Bacillota,and 3(5.17%)strains of Bacteroidota.From the isolated strains,18 petroleum hydrocarbon-degrading strains were screened out.[Conclusion]Through gradient domestication,seven natural microbial consortia were successfully enriched,achieving over 70%degradation of petroleum hydrocarbons at 7000 mg/L of 0#diesel.Amplicon sequencing revealed that varying 0#diesel concentrations altered the microbial consortium structure.Additionally,18 strains capable of using 0#diesel as the sole carbon source were identified,providing potential microbial resources for the bioremediation of oil-contaminated soil.
作者
独学渊
李书鹏
邱景琮
郭丽莉
常慧
李嘉晨
张冉
李丽杰
DU Xueyuan;LI Shupeng;QIU Jingcong;GUO Lili;CHANG Hui;LI Jiachen;ZHANG Ran;LI Lijie(National Engineering Laboratory for Site Remediation Technologies,Beijing,China;BCEG Environmental Remediation Co.,Ltd.,Beijing,China)
出处
《微生物学报》
北大核心
2025年第8期3794-3812,共19页
Acta Microbiologica Sinica
基金
北京建工集团科技计划(SA10000000272023051)。
关键词
石油烃降解
梯度驯化
微生物群落
细菌分离
petroleum hydrocarbon degradation
gradient domestication
microbial consortium
bacterial isolation
