Institute of Microbiology, University of Innsbruck, Technikerstrasse 25, A-6020 Innsbruck, Austria.
Sci Total Environ. 2012 Apr 1;421-422:184-96. doi: 10.1016/j.scitotenv.2012.01.043. Epub 2012 Mar 3.
A polyphasic approach combining culture-based methods with molecular methods is useful to expand knowledge on microbial diversity in contaminated soil. Microbial diversity was examined in soil samples from a former industrial site in the European Alps (mainly used for aluminum production and heavily contaminated with petroleum hydrocarbons) by culture-dependent and culture-independent methods. The physiologically active eubacterial community, as revealed by fluorescence-in-situ-hybridization (FISH), accounted for 6.7% of the total (DAPI-stained) bacterial community. 4.4% and 2.0% of the DAPI-stained cells could be attributed to culturable, heterotrophic bacteria able to grow at 20°C and 10°C, respectively. The majority of culturable bacterial isolates (34/48) belonged to the Proteobacteria (with a predominance of Alphaproteobacteria and Gammaproteobacteria), while the remaining isolates were affiliated with the Actinobacteria, Cytophaga-Flavobacterium-Bacteroides and Firmicutes. A high fraction of the culturable, heterotrophic bacterial population was able to utilize hydrocarbons. Actinobacteria were the most versatile and efficient degraders of diesel oil, n-alkanes, phenol and PAHs. The bacterial 16S rRNA gene clone library contained 390 clones that grouped into 68 phylotypes related to the Proteobacteria, Bacteroidetes, Actinobacteria and Spirochaetes. The archaeal 16S rRNA gene library contained 202 clones and 15 phylotypes belonging to the phylum Euryarchaeota; sequences were closely related to those of methanogenic archaea of the orders Methanomicrobiales, Methanosarcinales, Methanobacteriales and Thermoplasmatales. A number of bacterial and archaeal phylotypes in the clone libraries shared high similarities with strains previously described to be involved in hydrocarbon biodegradation. Knowledge of the bacterial and archaeal diversity in the studied soil is important in order to get a better insight into the microbial structure of contaminated environments and to better exploit the bioremediation potential by identifying potential hydrocarbon degraders and consequently developing appropriate bioremediation strategies.
一种结合基于培养的方法和分子方法的多相方法有助于扩展对污染土壤中微生物多样性的认识。通过依赖培养和非依赖培养的方法，对阿尔卑斯山（主要用于铝生产，受石油烃严重污染）一个前工业遗址的土壤样本进行了微生物多样性检测。荧光原位杂交（FISH）显示的生理活性真细菌群落占总（DAPI 染色）细菌群落的 6.7%。可培养异养细菌占 DAPI 染色细胞的 4.4%和 2.0%，分别能够在 20°C 和 10°C 下生长。大多数可培养细菌分离株（34/48）属于变形菌门（以α变形菌门和γ变形菌门为主），其余分离株属于放线菌门、噬纤维菌-黄杆菌-拟杆菌门和厚壁菌门。可培养异养细菌种群的很大一部分能够利用烃类。放线菌是柴油、正烷烃、苯酚和多环芳烃最具通用性和高效的降解菌。细菌 16S rRNA 基因克隆文库包含 390 个克隆，分为 68 个与变形菌门、拟杆菌门、放线菌门和螺旋体门相关的型。古菌 16S rRNA 基因文库包含 202 个克隆和 15 个属于广古菌门的型；序列与甲烷微生物目、甲烷杆菌目、甲烷杆菌目和热原体目等产甲烷古菌密切相关。克隆文库中的一些细菌和古菌型与先前描述的参与烃类生物降解的菌株具有很高的相似性。了解研究土壤中的细菌和古菌多样性对于更好地了解污染环境中的微生物结构以及通过鉴定潜在的烃类降解菌并相应地开发适当的生物修复策略来更好地利用生物修复潜力非常重要。
