| Project/Area Number |
14560051
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Plant nutrition/Soil science
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| Research Institution | Nagoya University |
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Principal Investigator |
ASAKAWA Susumu Nagoya University, Graduate School of Bioagricultural Sciences, Associate Professor, 大学院・生命農学研究科, 助教授 (50335014)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | Paddy field soil / Methanogenic archaea / Denaturing gradient gel electrophoresis method / PCR / 16S rDNA / Analysis of microbial community / 変性剤濃度勾配ゲル電気泳動(DGGE)法 |
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| Research Abstract |
The objective of the present research project was to establish a denaturing gradient gel electrophoresis (DGGE) method for analyzing 16S rDNA of methanogenic archaeal community in paddy field soil. Five specific primers for 16S rDNA of methanogenic archaea, which were modified from the primers for archaea, were first evaluated by PCR and DGGE using genomic DNAs of 13 pure culture strains of methanogenic archaea. The DGGE analysis was possible with two primer pairs (0348aF-GC and 0691R;0357F-GC and 0691R) of the five pairs tested although 16S rDNA of some nonmethanogenic archaea was amplified with O348aF-GC and 0691R. These two primer pairs were further evaluated for use in analysis of methanogenic archaeal communities in soil of two paddy fields in Chikugo and Anjo, Japan. Good separation and quality of patterns were obtained in DGGE analysis with the both primer pairs. Totally 41 DNA fragments were excised from the DGGE gels and their sequences were determined. All fragments belonged to methanogenic archaea and belonged to Methanomicrobiales, Methanosarcinales and "Rice clusters". These results indicate that the procedure of DGGE analysis with the primer pair of 0357F-GC and 0691R is suitable for investigating methanogenic archaeal community in paddy field soil. Now we are analyzing seasonal dynamics of their communities in the above two fields by this method.
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