Newmark Civil Engineering Laboratory, MC250, 205 North Mathews Avenue, Urbana, IL 61801, USA.
Appl Environ Microbiol. 2010 Jul;76(13):4179-84. doi: 10.1128/AEM.00193-10. Epub 2010 May 7.
To better understand the influence of environmental conditions on the adsorption of extracellular chromosomal DNA and its availability for natural transformation, the amount and conformation of adsorbed DNA were monitored under different conditions in parallel with transformation assays using the soil bacterium Azotobacter vinelandii. DNA adsorption was monitored using the technique of quartz crystal microbalance with dissipation (QCM-D). Both silica and natural organic matter (NOM) surfaces were evaluated in solutions containing either 100 mM NaCl or 1 mM CaCl(2). The QCM-D data suggest that DNA adsorbed to silica surfaces has a more compact and rigid conformation in Ca(2+) solution than in Na(+) solution and that the reverse is true when DNA is adsorbed to NOM surfaces. While the amounts of DNA adsorbed on a silica surface were similar for Ca(2+) and Na(+) solutions, the amount of DNA adsorbed on an NOM-coated surface was higher in Ca(2+) solution than in Na(+) solution. Transformation frequencies for dissolved DNA and DNA adsorbed to silica and to NOM were 6 x 10(-5), 5 x 10(-5), and 2.5 x 10(-4), respectively. For NOM-coated surfaces, transformation frequencies from individual experiments were 2- to 50-fold higher in the presence of Ca(2+) than in the presence of Na(+). The results suggest that groundwater hardness (i.e., Ca(2+) concentration) will affect the amount of extracellular DNA adsorbed to the soil surface but that neither adsorption nor changes in the conformation of the adsorbed DNA will have a strong effect on the frequency of natural transformation of A. vinelandii.
为了更好地理解环境条件对细胞外染色体 DNA 吸附及其对自然转化可用性的影响,我们使用土壤细菌固氮菌(Azotobacter vinelandii)平行进行转化实验,同时监测不同条件下吸附 DNA 的数量和构象。我们使用石英晶体微天平耗散技术(QCM-D)监测 DNA 吸附。我们在含有 100mM NaCl 或 1mM CaCl2 的溶液中评估了二氧化硅和天然有机物(NOM)表面。QCM-D 数据表明,在 Ca2+溶液中,吸附在二氧化硅表面的 DNA 具有更紧凑和刚性的构象,而在 Na+溶液中则相反;当 DNA 吸附在 NOM 表面时则相反。虽然在 Ca2+和 Na+溶液中,吸附在二氧化硅表面的 DNA 量相似,但在 Ca2+溶液中吸附在 NOM 覆盖表面上的 DNA 量高于在 Na+溶液中。溶解 DNA 和吸附在二氧化硅和 NOM 上的 DNA 的转化频率分别为 6x10(-5)、5x10(-5)和 2.5x10(-4)。对于 NOM 覆盖的表面,在存在 Ca2+的情况下,单个实验的转化频率比在存在 Na+的情况下高 2 到 50 倍。结果表明,地下水硬度(即 Ca2+浓度)会影响吸附到土壤表面的细胞外 DNA 的量,但吸附和吸附 DNA 构象的变化不会对固氮菌自然转化的频率产生强烈影响。