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Functional Genetic Diversity and Culturability of Petroleum-Degrading Bacteria Isolated From Oil-Contaminated Soils

机译：石油污染土壤中石油降解细菌的功能遗传多样性和可培养性

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In this study, we compared the culturability of aerobic bacteria isolated from long-term oil-contaminated soils via enrichment and direct-plating methods; bacteria were cultured at 30°C or ambient temperatures. Two soil samples were collected from two sites in the Shengli oilfield located in Dongying, China. One sample (S0) was close to the outlet of an oil-production water treatment plant, and the other sample (S1) was located 500 m downstream of the outlet. In total, 595 bacterial isolates belonging to 56 genera were isolated, distributed in Actinobacteria, Firmicutes, Bacterioidetes, and Proteobacteria. It was interesting that Actinobacteria and Firmicutes were not detected from the 16S rRNA gene clone library. The results suggested the activation of rare species during culture. Using the enrichment method, 239 isolates (31 genera) and 96 (22 genera) isolates were obtained at ambient temperatures and 30°C, respectively, from S0 soil. Using the direct-plating method, 97 isolates (15 genera) and 163 isolates (20 genera) were obtained at ambient temperatures and 30°C, respectively, from two soils. Of the 595 isolates, 244 isolates (41.7% of total isolates) could degrade n-hexadecane. A greater number of alkane-degraders was isolated at ambient temperatures using the enrichment method, suggesting that this method could significantly improve bacterial culturability. Interestingly, the proportion of alkane degrading isolates was lower in the isolates obtained using enrichment method than that obtained using direct-plating methods. Considering the greater species diversity of isolates obtained via the enrichment method, this technique could be used to increase the diversity of the microbial consortia. Furthermore, phenol hydroxylase genes (pheN), medium-chain alkane monooxygenases genes (alkB and CYP153A), and long-chain alkane monooxygenase gene (almA) were detected in 60 isolates (11 genotypes), 91 isolates (27 genotypes) and 93 isolates (24 genotypes), and 34 isolates (14 genotypes), respectively. This study could provide new insights into microbial resources from oil fields or other environments, and this information will be beneficial for bioremediation of petroleum contamination and for other industrial applications.
机译：在这项研究中，我们比较了通过富集和直接电镀方法从长期被石油污染的土壤中分离出的好氧细菌的可培养性；在30℃或环境温度下培养细菌。从位于中国东营的胜利油田的两个地点采集了两个土壤样品。一个样品（S0）靠近采油水处理厂的出口，另一个样品（S1）位于出口下游500 m。总共分离出属于56属的595个细菌分离株，分布在放线菌属，硬毛菌属，拟杆菌属和变形杆菌属中。有趣的是，从16S rRNA基因克隆文库中未检测到放线菌和Firmicutes。结果表明在培养过程中稀有物种的活化。使用富集方法，分别在环境温度和30°C下从SO土壤中获得了239株（31属）和96株（22属）的菌株。使用直接铺板法，分别在环境温度和30°C下从两种土壤中获得了97株（15属）和163株（20属）。在595株菌株中，有244株（占总菌株的41.7％）可以降解正十六烷。使用富集方法在环境温度下分离出大量烷烃降解剂，这表明该方法可以显着提高细菌的可培养性。有趣的是，在使用富集方法获得的分离物中，烷烃降解分离物的比例要比使用直接铺板方法获得的分离物低。考虑到通过富集方法获得的分离株具有更大的物种多样性，该技术可用于增加微生物群落的多样性。此外，在60株（11基因型），91株（27基因型）和93株中检测到苯酚羟化酶基因（pheN），中链烷烃单加氧酶基因（alkB和CYP153A）和长链烷烃单加氧酶基因（almA）。 （24个基因型）和34个分离株（14个基因型）。这项研究可以为油田或其他环境中的微生物资源提供新的见解，并且该信息将对石油污染的生物修复和其他工业应用有益。

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