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Phylogenetic study on microbial communities at various habitats in paddy field ecosystem

稻田生态系统不同生境微生物群落系统发育研究

基本信息

批准号：
15380051
负责人：
KIMURA Makoto
金额：
$ 9.86万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

For elucidating the community structure of microorganisms in the paddy field ecosystem, DNA and RNA were extracted from the samples collected from various habitats such as floodwater, rice straw on and in the plow layer, and surface oxidized and reduced soil layers, and they were PCR-amplified with primers specific to eubacteria, eukaryotes, methanogens, methanotrophs, and ammonia oxidizers. PCR products were subjected to denaturing gradient gel electrophoresis (DGGE) to estimate the seasonal variations and the specificity of respective habitats with the subsequent determination of DNA sequence for characteristic DGGE bands.In general, microbial communities estimated from PCR-DGGE patterns were phylogenetically endemic at respective habitats with relatively small seasonal variations. Sequence analysis of characteristic DGGE bands at each habitat indicated the presence of endemic microorganisms representing the community, and the microbial habitats were grouped into floodwater and percolating water (aquatic habitat), rice straw, rice straw compost and plant debris (plant residue habitat), rice roots and microcrustaceans (symbiotic habitat).As eubacterial and methanogenic communities in the plow layer soil that were observed in DGGE analysis of extracted DNA were also detected in DGGE analysis of extracted RNA, all the eubacteria and methanogens were estimated to be active all the year round.Interesting was the finding that microbial communities estimated from the PCR-DGGE analysis of environmental DNA was similar to those estimated from phospholipid fatty acid (PLFA) analysis.

为阐明稻田生态系统中微生物的群落结构，从不同生境（如洪水、耕层上和耕层中的稻草、表面氧化和还原土层）中采集样品，提取DNA和RNA，用真细菌、真核生物、产甲烷菌、甲烷氧化菌和氨氧化菌的特异性引物进行PCR扩增。PCR产物经变性梯度凝胶电泳（DGGE）分析，结果表明，PCR-DGGE图谱所显示的微生物群落在各生境中均具有明显的季节性特征，且季节性变化相对较小。DGGE序列分析表明，各生境中存在代表该群落的特有微生物，并将微生物生境分为洪水生境和淡水生境（水生生境）、稻草、稻草堆肥和植物残体（植物残体生境），水稻根和甲壳类由于在提取的DNA的DGGE分析中观察到的耕层土壤中的真细菌和产甲烷菌群落也在提取的RNA的DGGE分析中检测到，结果表明，利用PCR-DGGE技术分析的微生物群落结构与利用磷脂脂肪酸（PLFA）技术分析的微生物群落结构基本一致。