Department of Biology, Appalachian State University, Boone, NC 28608, USA.
Environ Microbiol. 2011 Mar;13(3):589-603. doi: 10.1111/j.1462-2920.2010.02360.x. Epub 2010 Oct 26.
Measurements of dissolved, ascorbate-reducible and total Mn by ICP-OES revealed significantly higher concentrations during estuarine turbidity maxima (ETM) events, compared with non-events in the Columbia River. Most probable number (MPN) counts of Mn-oxidizing or Mn-reducing heterotrophs were not statistically different from that of other heterotrophs (10³ -10⁴ cells ml⁻¹) when grown in defined media, but counts of Mn oxidizers were significantly lower in nutrient-rich medium (13 cells ml⁻¹). MPN counts of Mn oxidizers were also significantly lower on Mn(III)-pyrophosphate and glycerol (21 cells ml⁻¹). Large numbers of Rhodobacter spp. were cultured from dilutions of 10⁻² to 10⁻⁵, and many of these were capable of Mn(III) oxidation. Up to c. 30% of the colonies tested LBB positive, and all 77 of the successfully sequenced LBB positive colonies (of varying morphology) yielded sequences related to Rhodobacter spp. qPCR indicated that a cluster of Rhodobacter isolates and closely related strains (95-99% identity) represented approximately 1-3% of the total Bacteria, consistent with clone library results. Copy numbers of SSU rRNA genes for either Rhodobacter spp. or Bacteria were four to eightfold greater during ETM events compared with non-events. Strains of a Shewanella sp. were retrieved from the highest dilutions (10⁻⁵) of Mn reducers, and were also capable of Mn oxidation. The SSU rRNA gene sequences from these strains shared a high identity score (98%) with sequences obtained in clone libraries. Our results support previous findings that ETMs are zones with high microbial activity. Results indicated that Shewanella and Rhodobacter species were present in environmentally relevant concentrations, and further demonstrated that a large proportion of culturable bacteria, including Shewanella and Rhodobacter spp., were capable of Mn cycling in vitro.
通过电感耦合等离子体发射光谱法(ICP-OES)测量溶解态、抗坏血酸还原态和总锰,发现与非河口浑浊事件相比,哥伦比亚河河口浑浊最大值(ETM)事件期间的浓度明显更高。在限定培养基中生长时,锰氧化或锰还原异养菌的最可能数(MPN)计数与其他异养菌(10³-10⁴个细胞/ml)没有统计学差异,但在富营养培养基中,锰氧化菌的计数明显较低(13 个细胞/ml)。在焦磷酸锰和甘油上,锰氧化菌的 MPN 计数也明显较低(21 个细胞/ml)。从 10⁻²到 10⁻⁵的稀释度中培养了大量的 Rhodobacter spp.,其中许多能够氧化 Mn(III)。多达 30%的测试菌落呈 LBB 阳性,所有 77 个成功测序的 LBB 阳性菌落(形态各异)均产生与 Rhodobacter spp.相关的序列。qPCR 表明,一组 Rhodobacter 分离株和密切相关的菌株(95-99%的同一性)代表了大约 1-3%的总细菌,与克隆文库结果一致。在 ETM 事件期间,Rhodobacter spp.或细菌的 16S rRNA 基因的拷贝数比非 ETM 事件高 4-8 倍。从锰还原剂的最高稀释度(10⁻⁵)中回收了 Shewanella sp.的菌株,这些菌株也能够进行锰氧化。这些菌株的 16S rRNA 基因序列与克隆文库中获得的序列具有很高的同一性得分(98%)。我们的结果支持先前的发现,即 ETM 是微生物活性高的区域。结果表明,Shewanella 和 Rhodobacter 属以环境相关浓度存在,并且进一步表明,可培养细菌的很大一部分,包括 Shewanella 和 Rhodobacter spp.,能够在体外进行锰循环。