Analysis of the population dynamics of methanogens and sulfate-reducers in aquatic sediments by nucleic acid probes

利用核酸探针分析水生沉积物中产甲烷菌和硫酸盐还原菌的种群动态

• 批准号: 09680509
• 负责人: TAKII Susumu
• 金额: $ 2.24万
• 依托单位: TOKYO METROPOLITAN UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680509/
• 关键词: Sediment; Methanogens; Sulfate-reducing bacteria; rRNA; Hybridization; ドット・ブロット・ハイブリダイゼーション; メタン生成細菌; 底沢堆積物; リボソームRNA; オリゴヌクレオチド・プローブ; ハイブリダイゼーション; メタン生成速度; 硫酸還元速度; 底泥堆積物

We improved the hydroxyapatite spin column method to extract nucleic acids from aquatic sediments with low biological activities. On the rRNA extracted by this method from sediments of Lakes Kizaki and Biwa, and Tokyo Bay quantitative dot-blot hybridization was carried out using several oligonucleotide probes and the results were compared with metabolic activities such as methane production and sulfate reduction. The relative abundance of rRNA of Desulfobulbus was high compared with other sulfate-reducing bacteria (SRB) and correlated with the rate of sulfate reduction in sediment of L. Kizaki. Therefore, Desulfobulbus appears to be dominant in the active SRB population there. In Tokyo Bay Desulfobacter was a dominant SRB genus at a dredged site where the bottom water tended to be stagnant, while the generic composition of SRB was complex at a natural sea bed. At the former site the rate of sulfate reduction did not correlate with the relative abundance of Desulfobocter rRNA, but did with its ATP amount which calculated from the product of total ATP content in sediment and its relative RNA abundance. In the sediment of L. Biwa, the relative RNA abundances of Desulfobulbus and Desulfobacterium increased in winter when sulfate reduction rate was relatively high. The relative RNA abundance of archaea containing methanogens increased with depth of the sediment, but was low from 1.3 to 3.0%. The hybridization signals with probes targeting genera of methanogens were too week to be detected.

我们改进了羟基磷灰石自旋柱法，从生物活性较低的水生沉积物中提取核酸。用该方法从日本奇崎湖、琵琶湖和东京湾的沉积物中提取rRNA，用几种寡核苷酸探针进行定量斑点杂交，并将结果与甲烷产生和硫酸盐还原等代谢活动进行比较。与其他硫酸盐还原细菌(SRB)相比，Desulfobulbus的rRNA相对丰度较高，且与L.Kizaki沉积物中硫酸盐还原速率相关。因此，在那里活跃的SRB种群中，Desulfobulbus似乎是优势种。在东京湾疏浚底水趋于停滞的海域，硫杆菌属是主要的硫杆菌属，而在天然海床，硫杆菌属组成复杂。在前者，硫酸盐还原速率与脱硫菌rRNA的相对丰度无关，但与沉积物中总的ATP含量及其相对RNA丰度的乘积计算的ATP量有关。Biwa底泥中硫酸盐还原速率较高的冬季，硫酸盐还原菌和硫杆菌的相对RNA丰度较高。含产甲烷菌的古细菌的相对RNA丰度随着沉积物深度的增加而增加，但相对丰度较低，为1.3%~3.0%。针对产甲烷菌属的探针杂交信号太弱，检测不到。

项目成果

Purdy,K.J. et al.: "Use of 16S rRNA-targeted oligonucleotide probes to investigate occurrence and selection of sulphate-reducing bacteria in response to nutrient addition to sediment slurry microcosms from a Japanese estuary"FEMS Microbiology Ecol.ogy. 24

珀迪，K.J.

Takii, S. et al.: "Methane production and sulfate reduction in profundal sediments in Lake Kizaki, Japan"The Japanese Journal of Limnology. 58. 373-383 (1997)

Takii, S. 等人：“日本木崎湖深部沉积物中的甲烷产生和硫酸盐还原”《日本湖沼学杂志》。

Li, J-h., Purdy, K.J., Takii, S., Hayashi, H.: "Seasonal changes in ribosomal RNA of sulfate-reducing bacteria and sulfate reducing activity in a freshwater lake sediment" FEMS Microbiology Ecology. 28・1. 31-39 (1999)

Li，J-h.，Purdy，K.J.，Takii，S.，Hayashi，H.：“淡水湖沉积物中硫酸盐还原细菌的核糖体 RNA 和硫酸盐还原活性的季节性变化”FEMS 微生物学生态学 28・1-。 39 (1999)

Takii,S. et al.: "Methane production and sulfate reduction in profundal sediments in Lake Kizaki, Japan."The Japanese. Journal of Limnology. 58. 373-383 (1997)

泷井，S.

Li, J-H. et al.: "Seasonal changes in ribosomal RNA of sulfate-reducing bacteria and sulfate reducing activity in a freshwater lake sediment"FEMS Microbiology Ecol.ogy. 28. 31-39 (1999)

李，J-H。