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Organic chemial approach to the elucidation of the mechanism of plant transformation induced by Agrobacterium tumefaciens

有机化学方法阐明根癌农杆菌诱导植物转化的机制

基本信息

批准号：
03453145
负责人：
KAWAZU Kazuyoshi
金额：
$ 4.42万
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 (B)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1993
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03453145/
关键词：
Regulators of plant transfromation Agrobacterium tumefaciens beta-glucuronidase tabacco BY-2 suspension culture potato disk assay Ageratum oxazolomycin curromycin B curromycin B diacetate GUS遺伝子タバコ培養細胞BY-2 Julimycin B-II Oxazolomy

项目摘要

Plant transformation by Agrobacterium tumefaciens has been dramatically developed to create plants acquiring new characters, e.g., herbicide- or virus-resistance. Many scientists have been exploring the mechanism for the plant transformation, although the entire mechanism is not yet fully understood. As the plant transformation consists of several complex steps, a specific chemical probe affecting each step is required to elucidate its mechanism. The discovery of transformation regulators and the classification of these probes based on their mode of action are both helpful. We devised a rapid and quantitative bioassay method for regulators of plant tranformation by A.tumefaciens with an intron-GUS binary vector as an alternative method to the time-consuming potato tuber disk assay. Among several host plants tested, tobacco BY-2 suspension cells were the most preferable for this GUS expression assay. Furthermore, as the phytotoxicity of the test material, if any, is apparent from the growth inhibition of BY-2 suspension cells, this method can exclude phytotoxic compounds.In the course of our screening by the potato tuber disk assay for inhibitors of crown gall formation, oxazolomycin was isolated. Oxazolomycin inhibited crown gall formation owing to strong anitbacterial activity to the pathogen, A.tumefaciens. Interestingly, oxazolomycin esters had no antibacterial activity, although they inhibited crown gall formation. Curromycins, oxazolomycin analogs, were reproted as antibiotics. We isolated curromycin B from the curromycin-producing strain and prepared curromycin B diacetate. Curromycin B showed both crown gall formation inhibitory and antibacterial activities against A.tumefaciens. Curromycin B diacetate did not show any antibacterial activity, although it inhibited crown gall fromation. These results indicated that curromycin B diacetate was a promising chemical probe for studying the mechanism of plant transformation by A.tumefaciens.

通过根癌农杆菌进行的植物转化已经被显著地发展以产生获得新性状的植物，例如，抗除草剂或抗病毒。许多科学家一直在探索植物转化的机制，尽管整个机制尚未完全理解。由于植物转化由几个复杂的步骤组成，因此需要影响每个步骤的特定化学探针来阐明其机制。转化调节剂的发现和基于其作用模式的这些探针的分类都是有帮助的。我们设计了一种快速和定量的生物测定方法，植物转化调节剂的根癌农杆菌与内含子-GUS双元载体作为一种替代方法，费时的马铃薯块茎圆盘法。在所测试的几种宿主植物中，烟草BY-2悬浮细胞对于该GUS表达测定是最优选的。此外，由于试验材料的植物毒性（如果有的话）从BY-2悬浮细胞的生长抑制中是明显的，因此该方法可以排除植物毒性化合物。恶唑霉素抑制冠瘿的形成，由于强大的抗菌活性的病原菌，根癌农杆菌。有趣的是，恶唑霉素酯没有抗菌活性，虽然他们抑制冠瘿形成。Curromycins，恶唑霉素类似物，被认为是抗生素。从库罗霉素产生菌中分离得到库罗霉素B，并制备了库罗霉素B二乙酸酯。Curromycin B对根癌农杆菌表现出冠瘿形成抑制和抗菌活性。Curromycin B diacetate没有显示任何抗菌活性，虽然它抑制冠瘿的形成。这些结果表明，curromycin B diacetate是一种很有前途的化学探针，用于研究根癌农杆菌转化植物的机制。