Sahm K, MacGregor B J, Jørgensen B B, Stahl D A
Max Planck Institute for Marine Microbiology, Bremen, Germany.
Environ Microbiol. 1999 Feb;1(1):65-74. doi: 10.1046/j.1462-2920.1999.00007.x.
In the past, enumeration of sulphate-reducing bacteria (SRB) by cultivation-based methods generally contradicted measurements of sulphate reduction, suggesting unrealistically high respiration rates per cell. Here, we report evidence that quantification of SRB rRNA by slot-blot hybridization is a valuable tool for a more realistic assessment of SRB abundance in the natural environment. The distribution of SRB was investigated in a coastal marine sediment by hybridization of membrane-immobilized rRNA with oligonucleotide probes. As represented by general probe-target groups, SRB rRNA contributed between 18% and 25% to the prokaryotic rRNA pool. The dominant SRB were related to complete oxidizing genera (Desulphococcus, Desulphosarcina and Desulphobacterium), while Desulphobacter could not be detected. The vertical profile and quantity of rRNA from SRB was compared with sulphate reduction rates (SRR) measured with 35SO4(2-) tracer in whole-core incubations. While SRB abundance was highest near the surface, peaking at around 1.5 cm, measured sulphate reduction rates were lowest in this region. A second peak of SRB rRNA was observed at the transition zone from oxidized to reduced sediment, directly above the sulphate reduction maximum. Cell numbers calculated by converting the relative contribution of SRB rRNA to the percentage of DAPI-stained cells indicated a population size for SRB of 2.4-6.1 x 10(8) cells cm(-3) wet sediment. Cellular sulphate reduction rates calculated on the basis of these estimated cell numbers were between 0.01 and 0.09 fmol SO4(2-) cell(-1) day(-1), which is below the rates that have been determined for pure cultures (0.2-50 fmol SO4(2-) cell(-1) day(-1)) growing exponentially at nearoptimal temperature with a surplus of substrates.
过去,通过基于培养的方法对硫酸盐还原菌(SRB)进行计数,所得结果通常与硫酸盐还原的测量值相矛盾,这表明每个细胞的呼吸速率高得不切实际。在此,我们报告证据表明,通过狭缝印迹杂交对SRB rRNA进行定量,是更实际评估自然环境中SRB丰度的一个有价值的工具。通过将膜固定的rRNA与寡核苷酸探针杂交,研究了沿海海洋沉积物中SRB的分布。以一般探针 - 靶标组表示,SRB rRNA在原核生物rRNA池中占18%至25%。占主导地位的SRB与完全氧化的属(脱硫球菌属、脱硫八叠球菌属和脱硫杆菌属)有关,而未检测到脱硫杆菌属。将SRB的rRNA垂直分布和数量与在全岩芯培养中用35SO4(2-)示踪剂测量的硫酸盐还原速率(SRR)进行了比较。虽然SRB丰度在地表附近最高,在约1.5厘米处达到峰值,但该区域测量的硫酸盐还原速率最低。在从氧化沉积物到还原沉积物的过渡区,即在硫酸盐还原最大值正上方,观察到SRB rRNA的第二个峰值。通过将SRB rRNA的相对贡献转换为DAPI染色细胞的百分比来计算细胞数量,结果表明SRB的种群大小为2.4 - 6.1×10(8)个细胞/立方厘米湿沉积物。根据这些估计的细胞数量计算出的细胞硫酸盐还原速率在0.01至0.09飞摩尔SO4(2-)/细胞(-1)天(-1)之间,低于在接近最佳温度且底物过剩的情况下指数生长的纯培养物所确定的速率(0.2 - 50飞摩尔SO4(2-)/细胞(-1)天(-1))。