Methanogenic archaeal communities developed in paddy fields in the Kojima Bay polder, estimated by denaturing gradient gel electrophoresis, real-time PCR and sequencing analyses
Methanogenic archaeal communities developed in paddy fields in the Kojima Bay polder, estimated by denaturing gradient gel electrophoresis, real-time PCR and sequencing analyses
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DOI：
10.1111/j.1747-0765.2008.00334.x
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发表时间：
2009-02
影响因子：
2
通讯作者：
Takeshi Watanabe;V. Cahyani;J. Murase;E. Ishibashi;M. Kimura;S. Asakawa
中科院分区：
文献类型：
--
作者：
Takeshi Watanabe;V. Cahyani;J. Murase;E. Ishibashi;M. Kimura;S. Asakawa
摘要采用聚合酶链式反应-变性梯度凝胶电泳法、实时荧光定量聚合酶链式反应和测序技术对小岛湾围塘稻田土壤中产甲烷古菌群落进行了研究。2006年，从1692年至1954年开垦的四块稻田采集了耕层和底土层的土壤样本。从样品中提取的DNA扩增出的目标16S rRNA基因的DGGE带型与洪积和冲积区稻田土壤中的带型不同。与产甲烷古生菌和其他古生菌序列相关的靶向16SRRNA基因的数量在耕层和下层分别约为107g-108g和106g−-1干土。从主要的DGGE条带中获得了产甲烷古生菌的16S rRNA基因序列，它们分别属于甲烷菌目(稻米簇I)、甲烷琥珀酸目和甲烷杆菌目。而在洪积和冲积稻田中占主导地位的甲烷微生物群的序列没有被恢复。已知的嗜盐产甲烷菌和甲基化产甲烷菌是盐碱和海洋环境的特征，没有检测到。这些结果表明，小岛湾滩涂稻田土壤中发育了独特的产甲烷古生菌群落。
Abstract Methanogenic archaeal communities inhabiting the paddy field soils in the Kojima Bay polder were investigated using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), real-time PCR and sequencing analyses. Soil samples of the plow and subsoil layers were collected in 2006 from four paddy fields that were reclaimed between 1692 and 1954. The DGGE band patterns of the targeted 16S rRNA genes amplified from the extracted DNA from the samples were different from the patterns from the paddy field soils in diluvial and alluvial areas. The numbers of targeted 16S rRNA genes, which were involved with methanogenic archaeal and other archaeal sequences, were approximately 107–108 and 106 g−1 dry soil in the plow and subsoil layers, respectively. Sequences of methanogenic archaeal 16S rRNA genes belonging to Methanocellales (Rice cluster I), Methanosarcinales and Methanobacteriales were obtained from the major DGGE bands. Whereas sequences in Methanomicrobiales, which were predominant methanogens in the diluvial and alluvial paddy fields, were not recovered. Known halophilic and methylotrophic methanogens, which are characteristic of saline and marine environments, were not detected. These results indicate that distinctive methanogenic archaeal communities have developed in the paddy field soils in the Kojima Bay polder.