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Molecular biological analysis of the nitrogen-fixation ability of Sphingomonas paucimobilis

少动鞘氨醇单胞菌固氮能力的分子生物学分析

基本信息

批准号：
13660082
负责人：
HIDAKA Makoto
金额：
$ 0.51万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

Modern agriculture relies on large amount of chemical nitrogen fertilizer. This dependence has led to environmental eutorophicaton and consumption of much fossil fuel which causes environmental impact such as the promotion of global warming. To improve this tendency of modern agriculture, we are planning the continuous supply of nitrogen source in the field by taking advantage of biological nitrogen fixation. A diazotroph in genus Sphingomonas (Sphingomonas paucimobilis Y39) was isolated form rice rhizosphere. Afterward, it was discovered that may Sphingomonas bacteria are present on the surface of rice leaves or in rice plants. I expected that S.paucimobilis Y39 could be an endophyte of rice and provide efficient nitrogen to rice from both sides of the rice rhizosphexe and in plant by biological nitrogen fixation.At present, Sphingomonas paucimobilis Y39 was renamed Sphinomonas azotifigens Y39. At the beginning, I aimed to introduce the GFP gene into S.azotifigens Y39 for the purpose of demonstration of its presence in rice plants, i.e. demonstration of endophytic ability of S.azotifigens Y39. However, transformation of this bacterium was difficult and I have not obtained a GFP-expressing transformant. Thus, I started to isolate other endophytic Sphingomonas diazotrophs from flag leaves of rice, expecting that the obtained Sphingomonas bacteria will be easy to introduce the GEP gene. However, I have not yet isolated Sphingomonas diazotrophs.

现代农业依赖于大量的化学氮肥。这种依赖导致了环境污染和大量化石燃料的消耗，从而造成了环境影响，例如促进了全球变暖。为了改善现代农业的这一趋势，我们正在计划利用生物固氮的优势，在田间持续供应氮源。从水稻根际分离到一株鞘氨醇单胞菌(Sphingomonas Paucimobilis Y39)。后来，人们发现可能的鞘氨醇单胞菌存在于水稻叶片表面或水稻植株中。本研究认为Y39是一种水稻内生菌，可以通过生物固氮从水稻根茎两侧和植株内为水稻提供高效氮素。目前，该菌被重新命名为偶氮鞘氨醇单胞菌Y39。本研究首先将绿色荧光蛋白基因导入固氮链霉菌Y39，以证明其在水稻植株中的存在，即证明固氮链霉菌Y39的内生能力。然而，这种细菌的转化很困难，我还没有得到一个表达GFP的转化子。因此，我开始从水稻旗叶中分离出其他内生重氮鞘氨醇单胞菌，以期获得的鞘氨醇单胞菌能够容易地导入GEP基因。然而，我还没有分离出重氮鞘氨醇单胞菌。