CONSTRUCTION OF THE DEEP-SEA MICROBIAL IN SITU INCUBATION SYSTEM TO SEARCH FOR DIVERSIFIED DEEP-SEA BAROPHILIC MICROORGANISMS

建设深海微生物原位培养系统，寻找多样化的深海嗜压微生物

• 批准号: 07556139
• 负责人: OHWADA Kouichi
• 金额: $ 1.98万
• 依托单位: UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07556139/
• 关键词: barophile; marine microorganism; deep-sea bacteria; Deep-Sea Microbial in situ Incubation System; morring; pressurized culture; ZoBell 2216E; growth; 好圧性; 好圧性微生物; 深海; 多様性

The Deep-Sea Microbial in situ Incubation System was constructed with the following specificaton ;Maximum depth of use : 6,000 m Maximum duration of mooring : 1 year Weight and size : as small and light as possible so that submersible can bring to the deep-sea floor Operation : battery and timer Water sampling : tubing pump system (Yamato, Co., Japan) Incubators : 8 sets of sterile " Blood Bag System " (TERFLEX,Japan) with volume of 150 ml eachInto the 8 sets of incubators, small volume of the different substrates were added beforehand, and all the tubes from the incubators were connected to the tube pumping systems. The timer was set to start operation one day after the mooring to the deep-ocean. It was constructed that the battery inside the pressure tolerant battery house operated by a timer forces the motor to rotate which makes the cutter to rotate and cut off all the intake tubes to incubators and makes the tube pumping systems connected to the incubators to take ambient water in … More to the incubators.The Deep-Sea Microbial in situ Incubation System was moored at the site of the Japan Trench at a depth of 5,000 m on November 21,1996, at 38ﾟ01'N,145ﾟ32'E with several sets of water current meters at different depths during R/V Hakuho-maru cruise. On September 2,1997, it was successfully retrieved on board the vessel during cruise by R/V Tansei-maru after setting it for about 9 months. Whole set of incubation system was safe. Unfortunately, however, it was found that in the 8 sets of tubing pumping systems only one worked well and other 7 sets didn't take any deep-sea water sample at all into incubation bags. In one incubation bag in which ZoBell 2216 E was added as a substrate, dense growth of deep-sea bacteria (1.1x10^8 cells/ml) was observed. Experimental culture of them under 1 atm condition did not show any growth at all. On the contrary they showed good growth under pressurized condition (500 atm) at 2 C.Isolation of these bacterial strains and study for phylogeny of them are now undertaken. Less

深海微生物原位培养系统按照以下规格建造;最大使用深度：用途：6，000 m最长系泊时间：1年重量和尺寸：尽可能小和轻，以便潜水器可以带到深海海底操作：电池和计时器水取样：管道泵系统（Yamato，Co.，日本）培养箱：8套无菌的“Blood Bag System“（TERFLEX，日本），每一套的体积为150 ml。在8套培养箱中，预先加入小体积的不同底物，并将来自培养箱的所有管连接到管泵送系统。计时器被设置为在深海系泊后一天开始运行。其构造为，由定时器操作的耐压电池室内的电池迫使马达旋转，这使得切割器旋转并切断所有通向培养箱的进水管，并使得连接到培养箱的管泵送系统将环境水吸入 ...更多信息 深海微生物原位培养系统于1996年11月21日停泊在日本海沟5，000 m深的地点，北纬38度01分，东经145度32分，在“白凤丸”号考察船航行期间，在不同深度安装了几套水流计。1997年9月2日，在放置了约9个月后，由R/V Tansei Maru在巡航期间成功地将其打捞到船上。整套孵化系统是安全的。然而，不幸的是，在8套管道泵系统中，发现只有一套工作良好，其他7套根本没有将任何深海水样品放入培养袋中。在一个添加ZoBell 2216 E作为底物的培养袋中，观察到深海细菌密集生长（1.1x10^8个细胞/ml）。在1个大气压条件下进行的实验培养根本没有显示出任何生长。相反，它们在2 ℃的加压条件（500 atm）下显示出良好的生长。现在正在进行这些细菌菌株的分离和它们的繁殖研究。少

项目成果

崎山徳起.大和田紘一: "Isolation and groioth charaete ristice of deep-sea barophilic baeteua from the Japan Trench." Fisheries Science. 63. 228-232 (1997)

Tokuoki Sakiyama 和 Koichi Owada：“日本海沟深海嗜压细菌的分离和生长特征”，63. 228-232 (1997)。

崎山 徳起・大和田 絋一: "Effect of hydrostatic pressure on the groeuet of deep-sea bacteriel communities." Proceedings NIPR Symp.Polar Biology. 12. 印刷中 (1998)

Tokioki Sakiyama 和 Keiichi Owada：“静水压力对深海细菌群落的影响。”Proceedings NIPR Symp.Polar Biology 12。出版中（1998 年）。

大和田紘一: "バイオマーカーによる海洋微生物群集の解析" 月刊 海洋. 29. 707-709 (1997)

Koichi Owada：“利用生物标志物分析海洋微生物群落”，《海洋月刊》29. 707-709 (1997)。

崎山徳起,大和田紘一: "Isolation and Growth Characteristies of Deep-Sea Barophilec Bacteria from the Japan Trench" Fisheries Science. 63・2(印刷中). (1997)

崎山德兴、小和田浩一：“日本海沟深海嗜压细菌的分离和生长特征”渔业科学 63・2（出版中）。

深川 秀敏・塚本 久美子・大和田 絋一: "Restriction fragment lergil polymorphism areelepis of 16SrRNA genes of the family Vibrionaceal isolated from cold marine enuironments." Proceedings NIPR Symp.Polar Biology. 12. 印刷中 (1998)

Hidetoshi Fukakawa、Kumiko Tsukamoto、Keiichi Owada：“从寒冷海洋环境中分离出的弧菌科 16SrRNA 基因的限制性片段 lergil 多态性。” NIPR Symp.Polar Biology 12 论文集（1998 年）。