Borin Sara, Crotti Elena, Mapelli Francesca, Tamagnini Isabella, Corselli Cesare, Daffonchio Daniele
University of Milan, Department of Food Science and Microbiology, Milan, Italy.
Saline Syst. 2008 Aug 5;4:10. doi: 10.1186/1746-1448-4-10.
Extracellular dissolved DNA has been demonstrated to be present in many terrestrial and aquatic environments, actively secreted, or released by decaying cells. Free DNA has the genetic potential to be acquired by living competent cells by horizontal gene transfer mediated by natural transformation. The aim of this work is to study the persistence of extracellular DNA and its biological transforming activity in extreme environments like the deep hypersaline anoxic lakes of the Mediterranean Sea. The brine lakes are separated from the upper seawater by a steep chemocline inhabited by stratified prokaryotic networks, where cells sinking through the depth profile encounter increasing salinity values and osmotic stress.
Seven strains belonging to different taxonomic groups isolated from the seawater-brine interface of four hypersaline lakes were grown at medium salinity and then incubated in the brines. The osmotic stress induced the death of all the inoculated cells in variable time periods, between 2 hours and 144 days, depending on the type of brine rather than the taxonomic group of the strains, i.e. Bacillaceae or gamma-proteobacteria. The Discovery lake confirmed to be the most aggressive environment toward living cells. In all the brines and in deep seawater dissolved plasmid DNA was substantially preserved for a period of 32 days in axenic conditions. L'Atalante and Bannock brines induced a decrease of the supercoiled form up to 70 and 40% respectively; in the other brines only minor changes in plasmid conformation were observed. Plasmid DNA after incubation in the brines maintained the capacity to transform naturally competent cells of Acinetobacter baylii strain BD413.
Free dissolved DNA is likely to be released by the lysis of cells induced by osmotic stress in the deep hypersaline anoxic lakes. Naked DNA was demonstrated to be preserved and biologically active in these extreme environments, and hence could constitute a genetic reservoir of traits acquirable by horizontal gene transfer.
细胞外溶解DNA已被证明存在于许多陆地和水生环境中,它可由活细胞主动分泌,也可因细胞腐烂而释放。游离DNA具有通过自然转化介导的水平基因转移被有感受态的活细胞获取的遗传潜力。本研究的目的是探究在地中海深层高盐缺氧湖泊等极端环境中细胞外DNA的持久性及其生物转化活性。这些盐湖通过一个陡峭的化学分层与上层海水隔开,化学分层中存在分层的原核生物网络,细胞在下沉过程中会遇到盐度不断增加和渗透压不断增大的情况。
从四个高盐湖泊的海水-盐水界面分离出的七株属于不同分类群的菌株,先在中等盐度下培养,然后在盐水中孵育。渗透压在2小时至144天的不同时间段内导致所有接种细胞死亡,这取决于盐水的类型而非菌株的分类群,即芽孢杆菌科或γ-变形菌纲。发现湖被证实对活细胞的侵蚀性最强。在所有盐水中以及深海海水中,无菌条件下溶解的质粒DNA在32天内基本保持完整。阿塔兰特和班诺克盐水分别导致超螺旋形式减少高达70%和40%;在其他盐水中,仅观察到质粒构象有微小变化。在盐水中孵育后的质粒DNA仍保持转化鲍氏不动杆菌BD413菌株天然感受态细胞的能力。
在深层高盐缺氧湖泊中,游离溶解DNA可能是由渗透压诱导的细胞裂解释放出来的。在这些极端环境中,裸露的DNA被证明是可以保存且具有生物活性的,因此可能构成一个可通过水平基因转移获取性状的遗传库。