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The quantitative evaluation offixed nitrogen by the endophytic bacteria in sweet potato and sugarcane

甘薯和甘蔗内生细菌固定氮的定量评价

基本信息

批准号：
16380053
负责人：
AKAO Shoichiro
金额：
$ 7.01万
依托单位：
University of Miyazaki
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2007
项目状态：
已结题

项目摘要

The nitrogen-fixing bacteria which inhabits the inside plant is being called nitrogen-fixation endophytic bacteria. The plant which fixes nitrogen is limited to the leguminous plants which have a symbiotic relationship with RhJzobia and about ten kinds of plants that have a symbiotic relationship with Frankia However, several twenty years ago ; the existence of the nitrogen-fixing endophitic bacteria was isolated from sugarcane in Brazil And also it was reported that the rate of fixed nitrogen to total nitrogen might reach 60-70% in the sugarcane that nitrogen- fixing endophytic bacteria inhabited. The rate of contribution of N2 fixation to total plant N is equal to the Rhizobium-legume symbiosis. As for the nitrogen fixing endophytic bacteria, they were isolated from wetland rice, wheat, maize, sorghum, and various forage grasses in addition to sugarcane. This showed the possibility that a nitrogen fixing function is given to the non-leguminous plants. But, there are some problems to … More solve to realize this. They were the selection of useful endophytic bacteria and a colonization to the inside plants. The infection and multiplication to the plant of the nitrogen fixing endophytic bacteria and the estimation method of the nitrogen fixed by endophytic diazotrophs was examined in this research.A research period is four years from the 16th year of Heisei until 19. At the first half of research period, the inoculation examination by GFP-labeled Herbaspirillum sp. was conducted. And the isolation of the endophytic diazotrophs, are responsible for the observed N2 fixation were tried from sugarcane and sweet potato. Moreover the gene encoding green fluorescent protein (GFP) via conjugation and electotroporation At the latter half of research period, inoculation effectiveness and internal colonization ability as an introduced in the non host plants of endophytic diazotrophs isolated from sugarcane and sweet potato was examined. And also the method for the amount of nitrogen fixed a period of duration was examined.As a result, we determined the relative abundance of N2-fixing enndophytic bacteria in the stem of sugarcane cvs. NiF8, Nil5, N7 and KTN94-88, and in the stem, tuber, leaf and root of sweet potato cvs. Koganesengan and Shiroyutaka and 39 isolates, 27 isolates from sugarcane and 12 isolates from sweet potato, were obtained from the above-mentioned materials. Herbaspirillum sp., Enterobacter sp. and Pantoea sp. used in this experiment could be established in plants other than their original host, suggesting a lack of host specificity, although the extent of colonization may vary depending on the combination of host and endophyte. And, higher bacterial numbers were detected in plant tissues inoculated with higher inoculum (10^8 cells/ml)concentration treatment.We have used the root part of the micropropagated sugarcane plant to the target mainly as an inoculation part of the useful N_2-fixing endophytic bacteria in this study. However we got the conclusion that a suitable delivery method for introducing endophytic bacteria into the vegetative propagation plants such as sugarcane is the vacuum infiltration method to stem pieces. Less

居住在植物内部的固氮细菌被称为固氮内生细菌。固氮植物仅限于与根瘤菌属有共生关系的豆科植物和与根瘤菌属有共生关系的十余种植物。从巴西甘蔗中分离到固氮内生细菌的存在，并报道固氮内生细菌所栖息的甘蔗中固氮占总氮的比例可达60-70%。固氮对植株总氮的贡献率与根瘤菌-豆科植物共生速率相等。固氮内生细菌除从甘蔗中分离到外，还从湿地水稻、小麦、玉米、高粱和各种牧草中分离到。这表明非豆科植物具有固氮功能的可能性。但是，要实现这一点，还有一些问题需要解决。它们是对有益内生细菌的选择和对植物内部的定植。研究了固氮内生细菌对植物的侵染和繁殖，以及内生重氮营养体固氮量的估算方法。研究时间为4年，从平成16年到19年。在研究的前半期，用gfp标记的Herbaspirillum sp.进行接种检查。并从甘蔗和甘薯中分离出了固氮的内生重氮营养菌。在研究后期，对甘蔗和甘薯内生重氮养菌的接种效果和内部定殖能力进行了检测。并对一段时间内固定氮量的方法进行了探讨。因此，我们测定了甘蔗茎秆中固氮内生细菌的相对丰度。NiF8、Nil5、N7和KTN94-88在甘薯茎、块茎、叶和根中的表达。从上述材料中分离得到Koganesengan和Shiroyutaka菌株39株，甘蔗27株，甘薯12株。本实验中使用的Herbaspirillum sp., Enterobacter sp.和Pantoea sp.可以在其原始宿主以外的植物中建立，这表明缺乏宿主特异性，尽管定植程度可能取决于宿主和内生菌的组合。较高接种量（10^8个细胞/ml）处理的植物组织中细菌数量较高。在本研究中，我们主要利用甘蔗植株的根部分作为接种有用的固氮内生细菌的部位。然而，我们得出结论，将内生细菌引入甘蔗等无性繁殖植物的一种合适的传递方法是真空渗透法。少

项目成果

期刊论文数量（35）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Detection of gfp expression from gfp-labelled bacteria spot inoculated onto sugarcane tissues

甘蔗组织上gfp标记细菌斑点接种后gfp表达的检测

DOI：
发表时间：
2005
期刊：
African Journal of Biotechnology Vol.4
影响因子：
0
作者：
J. Njoloma;M. Oota;Y. Saeki;S. Akao
通讯作者：
S. Akao

^<15>N同位体希釈法によるサトウキビの固定窒素量の推定

^<15>利用N同位素稀释法估算甘蔗中的固定氮量

DOI：
发表时间：
2005
期刊：
宮崎大学農学部研究報告 51(1・2)
影响因子：
0
作者：
西口;清水;ジョイス;大田;佐伯;赤尾
通讯作者：
赤尾

Colonization and growth promotion characteristics of Herbaspirillum sp.B501 and Enterobacter sp.for Brasica oleracea

甘蓝中Herbaspirillum sp.B501和Enterobacter sp.的定植和促生特性

DOI：
发表时间：
2007
期刊：
影响因子：
0
作者：
Zakria M.;大迫明日香・佐伯雄一・山本昭洋・赤尾勝一郎
通讯作者：
大迫明日香・佐伯雄一・山本昭洋・赤尾勝一郎

サトウキビに接種した窒素固定エンドファイトHerbaspirillumの動態追跡のための緑色蛍光タンパク質の利用

利用绿色荧光蛋白追踪接种甘蔗的固氮内生菌草螺菌的动态

DOI：
发表时间：
2006
期刊：
土と微生物 60
影响因子：
0
作者：
Njoloma;J. P.;M. Oota;K. Taroura;Y. Saeki;S. Akao;清水友・豊里学・久綱泰代・佐伯雄一・南澤究・中西康博・Joyce Nioloma・明石良・赤尾勝一郎
通讯作者：
清水友・豊里学・久綱泰代・佐伯雄一・南澤究・中西康博・Joyce Nioloma・明石良・赤尾勝一郎

N2 fixation in sugarcane and population of N2-fixing endophytes in stem apoplast solution

甘蔗中的 N2 固定以及茎质外体溶液中固氮内生菌的种群