Marcos M S, Lozada M, Dionisi H M
Environmental Microbiology Laboratory, Patagonian National Research Center (CENPAT - CONICET), Puerto Madryn (U9120ACF), Chubut, Argentina.
Lett Appl Microbiol. 2009 Nov;49(5):602-8. doi: 10.1111/j.1472-765X.2009.02711.x. Epub 2009 Aug 11.
The goal of this study was to identify functional targets to detect polycyclic aromatic hydrocarbon (PAH)-degrading bacterial populations in cold marine ecosystems.
We designed a degenerate primer set targeting genes encoding the alpha subunit of PAH-dioxygenases from Gram-positive bacteria. This primer set was used to amplify gene fragments from metagenomic DNA isolated from Subantarctic marine sediments (Ushuaia Bay, Argentina). These gene fragments were cloned and sequenced. We identified 14 distinct groups of genes, most of them showing significant relatedness with dioxygenases from Gram-positive bacteria of the genera Rhodococcus, Mycobacterium, Nocardioides, Terrabacter and Bacillus. The level of identity with these genes, however, was low to moderate (33-62% at the amino acid level).
These results indicate the presence of a high diversity of hitherto unidentified dioxygenase genes in this cold polluted environment.
Subantarctic marine ecosystems are particularly vulnerable to hydrocarbon pollution, and the development of environmental restoration strategies for these environments is pressing. The information obtained in this work will be the starting point for the design of quantitative molecular tools to analyse the abundance and dynamics of these aromatic hydrocarbon-degrading bacterial populations in the marine environment.
本研究的目的是确定功能性靶点,以检测寒冷海洋生态系统中多环芳烃(PAH)降解细菌种群。
我们设计了一套简并引物,靶向革兰氏阳性菌中编码PAH-双加氧酶α亚基的基因。该引物用于从南极海洋沉积物(阿根廷乌斯怀亚湾)分离的宏基因组DNA中扩增基因片段。这些基因片段被克隆并测序。我们鉴定出14个不同的基因群,其中大多数与红球菌属、分枝杆菌属、诺卡氏菌属、土杆菌属和芽孢杆菌属革兰氏阳性菌的双加氧酶有显著相关性。然而,与这些基因的同一性水平较低至中等(氨基酸水平为33%-62%)。
这些结果表明,在这个寒冷污染环境中存在高度多样性的迄今未鉴定的双加氧酶基因。
南极海洋生态系统特别容易受到碳氢化合物污染,为这些环境制定环境恢复策略迫在眉睫。本研究获得的信息将作为设计定量分子工具的起点,用于分析海洋环境中这些芳烃降解细菌种群的丰度和动态。