Sievert S M, Brinkhoff T, Muyzer G, Ziebis W, Kuever J
Department of Microbiology, Max-Planck-Institute for Marine Microbiology, D-28359 Bremen, Germany.
Appl Environ Microbiol. 1999 Sep;65(9):3834-42. doi: 10.1128/AEM.65.9.3834-3842.1999.
The spatial heterogeneity of bacterial populations at a shallow-water hydrothermal vent in the Aegean Sea close to the island of Milos (Greece) was examined at two different times by using acridine orange staining for total cell counts, cultivation-based techniques, and denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA gene fragments. Concurrent with measurements of geochemical parameters, samples were taken along a transect from the center of the vent to the surrounding area. Most-probable-number (MPN) counts of metabolically defined subpopulations generally constituted a minor fraction of the total cell counts; both counting procedures revealed the highest cell numbers in a transition zone from the strongly hydrothermally influenced sediments to normal sedimentary conditions. Total cell counts ranged from 3.2 x 10(5) cells ml(-1) in the water overlying the sediments to 6.4 x 10(8) cells g (wet weight) of sediment(-1). MPN counts of chemolithoautotrophic sulfur-oxidizing bacteria varied between undetectable and 1.4 x 10(6) cells g(-1). MPN counts for sulfate-reducing bacteria and dissimilatory iron-reducing bacteria ranged from 8 to 1.4 x 10(5) cells g(-1) and from undetectable to 1.4 x 10(6) cells g(-1), respectively. DGGE revealed a trend from a diverse range of bacterial populations which were present in approximately equal abundance in the transition zone to a community dominated by few populations close to the center of the vent. Temperature was found to be an important parameter in determining this trend. However, at one sampling time this trend was not discernible, possibly due to storm-induced disturbance of the upper sediment layers.
通过使用吖啶橙染色法进行总细胞计数、基于培养的技术以及对聚合酶链反应(PCR)扩增的16S rRNA基因片段进行变性梯度凝胶电泳(DGGE)分析,在两个不同时间对希腊米洛斯岛附近爱琴海一处浅水热液喷口处细菌种群的空间异质性进行了研究。在测量地球化学参数的同时,沿着从喷口中心到周边区域的一条样带采集了样本。代谢定义的亚种群的最大可能数(MPN)计数通常仅占总细胞计数的一小部分;两种计数方法均显示,在从受强烈热液影响的沉积物到正常沉积条件的过渡带中细胞数量最多。总细胞计数范围从沉积物上方水体中的3.2×10⁵个细胞/毫升到沉积物(湿重)中的6.4×10⁸个细胞/克。化能自养型硫氧化细菌的MPN计数在无法检测到至1.4×10⁶个细胞/克之间变化。硫酸盐还原细菌和异化铁还原细菌的MPN计数分别在8至1.4×10⁵个细胞/克以及无法检测到至1.4×10⁶个细胞/克之间。DGGE揭示了一种趋势,即从过渡带中存在的种类繁多且丰度大致相等的细菌种群,到喷口中心附近由少数种群主导的群落。温度被发现是决定这种趋势的一个重要参数。然而,在一次采样时这种趋势并不明显,可能是由于风暴对上覆沉积层的扰动所致。