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Chemical Studies on the Infection Mechanism of Crown Gall Disease

冠胆病感染机制的化学研究

基本信息

批准号：
01560152
负责人：
KAWAZU Kazuyoshi
金额：
$ 1.34万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01560152/
关键词：
Crown gall Potato disk method Plant transformation inhibition Julichrome Q1・3 Julimycine B-II Agrobacterium tumefaciens Octopine Nopaline クラウンゴ-ル形質転換阻害 Oxazolomycin JulichromeQ1・3 JulimycinBーII アントラキノン

项目摘要

1. Agrobacterium tumefaciens is a soil bacterium which induces crown gall tumors in a wide variety of dicotyledonous plants. This tumorigenic interaction between the bacterium and the plant is a natural example of the genetic engineering of eukaryotes. Indeed, the oncogenic bacterium harbors a large Ti plasmid, whose T-DNA region is transferred to and integrated into the plant genome during tumorous transformation. However, very little information is available concerning the early process leading to tumor formation. In order to study further the mechanism of tumor induction, it is necessary to develop a chemical probe that will help in exploring the early steps of tumor formation. We searched transformation inhibitors which do not inhibit the growth of A. tumefaciens and are nontoxic against the transformed plant cells.2. A few esters derived from oxazolomycin isolated from Streptomyces sp. KBFP-2025 as well as two active compounds of bisanthraquinone type, julichrome Q_<1・3> and julimycin B-II, isolated from Streptomyces sp. TM-71, inhibit crown gall formation, but are nontoxic against this bacterium and the host plant.3. These compounds administered to a potato disk 12 - 24 hours after the inoculation of the bacterium hardly inhibited crown gall formation, and no longer necrosed the potato disk. This suggests that these compounds act on any step of transformation process, but have no toxicity toward transformed plant cells whose genome has integrated the T-DNA of A. tumefaciens. It is probable that the transformation process of this system is completed within 12 - 24 hours after the inoculation. For a direct proof of this hypothesis, a quantitative microanalytical method of opines, indices of transformation, has to be established. This method involves HPLC analysis of fluorescent compound formed by reaction of benzoin with guanidino group of the characteristic amino acid. Time course of transformation will be possibly traced by this method.

1.根癌农杆菌是一种土壤细菌，可诱导多种双子叶植物的根癌瘤。细菌和植物之间的这种致瘤相互作用是真核生物基因工程的一个自然例子。事实上，致癌细菌携带一个大的Ti质粒，其T-DNA区域在肿瘤转化过程中转移并整合到植物基因组中。然而，关于导致肿瘤形成的早期过程的信息很少。为了进一步研究肿瘤诱导的机制，有必要开发一种有助于探索肿瘤形成早期步骤的化学探针。我们寻找不抑制A.对转化的植物细胞无毒.从链霉菌KBFP-2025中分离的恶唑霉素衍生的几种酯类化合物以及从链霉菌TM-71中分离的两种双蒽醌型活性化合物：尤利铬Q_<1·3>和尤利霉素B-II对冠瘿菌的形成有抑制作用，但对该菌和寄主植物无毒性.在接种细菌后12 - 24小时，将这些化合物施用于马铃薯圆片几乎不抑制冠瘿形成，并且不再使马铃薯圆片坏死。这表明，这些化合物作用于转化过程的任何步骤，但对基因组已整合A.根瘤菌该系统的转化过程可能在接种后12 - 24小时内完成。为了直接证明这一假设，必须建立一种观点的定量微观分析方法，即转化指数。该方法涉及苯偶姻与特征氨基酸的胍基反应形成的荧光化合物的HPLC分析。利用这种方法可以追踪到转化的时间过程。