ISOLATION AND CHARACTERIZATION OF EXTREMELY THERMOPHILIC ARCHAEBACTERIA RELATED TO THE GENUS THERMOCOCCUS FROM DEEP-SEA HYDROTHERMAL GUAYMAS BASIN
ISOLATION AND CHARACTERIZATION OF EXTREMELY THERMOPHILIC ARCHAEBACTERIA RELATED TO THE GENUS THERMOCOCCUS FROM DEEP-SEA HYDROTHERMAL GUAYMAS BASIN
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DOI：
10.1007/bf00293552
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发表时间：
1995-09-01
影响因子：
2.6
通讯作者：
BARBIER, G
中科院分区：
文献类型：
--
作者：
ANTOINE, E;GUEZENNEC, J;BARBIER, G
1991 年 11 月，IFREMER 进行的“Guaynaut”海洋学巡航期间，从瓜伊马斯盆地（加利福尼亚州中部海湾）活跃热液区的高温和富含石油的沉积物中收集了沉积物岩心。这些样本是由法国深海载人潜水器“鹦鹉螺号”在2000米深处采集的。在沉积物内的活跃吸烟者块体组合的底部钻了四个 20-40 厘米长的沉积岩芯，其顶部温度为 3.5 摄氏度，20 厘米深度为 105 摄氏度。接种到无硫 SME 液体培养基后，将它们分成 22 个 5 厘米厚的切片，用于分离异养超耐热微生物。从这些富集中获得了 18 个分离株，其中 2 个在 95 摄氏度下生长，16 个在 80 摄氏度下生长，并对其进行了分类学表征。脂质分析表明细胞壁和细胞膜中存在二醚和四醚，这是古细菌的特征。扫描电子显微镜检查显示，分离株是大小不一的球菌（直径0.1至0.5微米）或厚0.5微米、直径1微米的厚堆积圆盘。两种形式都嵌入密集的光纤网络中。在生理学上，它们被发现是厌氧、异养和超嗜热的（80°C-95°C）。 DNA 碱基组成的测定得出的 G + C mol % 值范围为 36 至 57。 18 个分离 DNA 与超耐热古细菌参考菌株 DNA 的定性杂交表明，既不与两种火球菌属杂交，也不与两种脱硫球菌属杂交，也不与海葡萄球菌杂交。另一方面，所有分离株均与所测试的三种热球菌属（T. celer、T. stetteri、T. litoralis）中的至少一种杂交。 rrn操纵子PCR扩增片段的限制性多态性显示，12个分离株与T. celer和T. stetteri具有相同的谱，4个分离株与T. litoralis具有相同的谱，2个具有新的谱，表明它们是新种。
During the ''Guaynaut'' oceanographic cruise performed by IFREMER in November 1991, sediment cores were collected from high-temperature and petroleum-rich deposits in an active hydrothermal zone, at the Guaymas basin (Central gulf of California). Those samples were collected by the French deep-sea manned submersible ''Nautile'' at a depth of 2000 meters. Four sediment cores of 20-40-cm length were drilled at the bottom of a block assemblage of active smokers inside sediments whose temperatures were 3.5 degrees C on the top to 105 degrees C at 20 cm depth. They were subsampled in 22 slices of 5-cm thickness and used for isolation of heterotrophic hyperthermophilic microorganisms, after inoculation in sulfur-free SME liquid medium. From those enrichments 18 isolates were obtained, 2 growing at 95 degrees C and 16 at 80 degrees C, and their taxonomic characterization was undertaken. Lipid analysis indicated the presence of diethers and tetraethers in the cell walls and membranes, characteristic of Archaebacteria. Examinations by scanning electron microscopy showed that isolates were cocci of heterogeneous sizes (diameter from 0.1 to 0.5 micrometers) or thick, piled-up discs 0.5 mu m thick and 1 mu m in diameter. Both forms were embedded in a dense fiber network. Physiologically they were found to be anaerobic, heterotrophic, and hyperthermophilic (80 degrees-95 degrees C). Determination of the DNA base composition resulted in G + C mol % values ranging from 36 to 57. Qualitative hybridizations of the 18 isolate DNAs with hyperthermophilic Archaebacteria reference strain DNAs showed that hybridizations occurred neither with the two species of Pyrococcus nor with the two species of Desulfurococcus, nor with Staphylothermus marinus. On the other hand, all isolates hybridized with at least one of the three species of Thermococcus tested (T. celer, T. stetteri, T. litoralis). Restriction polymorphism on a PCR-amplified fragment of the rrn operon showed that 12 isolates had the same profile as T. celer and T. stetteri, 4 isolates had the same profile as T. litoralis, and 2 had new profiles, suggestive that they are new species.