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Molecular mechanisms of T-DNA transfer from Agrobacterium to plant cells during formations of crown gas tumor.

冠气瘤形成过程中 T-DNA 从农杆菌转移到植物细胞的分子机制。

基本信息

批准号：
60480011
负责人：
MACHIDA Yasunori
金额：
$ 4.22万
依托单位：
Nagoyo Univetsity
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1987
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60480011/
关键词：
Agrobacterium Ti plasmid T-DNA Crown gal tumor Plants Monocotyledonous plants Vir gene DNA technology

项目摘要

We have demonstrated that when Agrobacterium infects dicotyledonous plants, the T-DNA region of Ti plasmid is circularized in bacterial cells. This reaction is induced by the diffusible plant factor(s) secreted from plant cells. We also have shown that monocotyledonous plants which have been thought to be resistant to agrobacterial infection contain inducing substance(s), yet it differs from that of dicotyledonous plants. We try to purify the plant factor from wheat bran and determine its molesular structures. During purification, inducing activity was split into two fractions, one was hydrophilic and the other hydrophobic, both of which were required for induction of T-DNA circularization. In addition, we have revealed that the virD locus of Ti plasmid is involved in the circularization event. The virD1 and D2 genes in this locus are together necessary. Products encoded by these genes seem to have activity of endonuclease or recombinase specific for the T-DNA border sequences. RMA polymerase of Agrobacterium was shown to be a negative regulator for T-DNA circularization which competes with products of virD1 and D2. We try to purify both virD proteins as well as other virD proteins (D3 and D4), and elucidate enzymatic properties of these proteins.

我们已经证明，当农杆菌侵染双子叶植物时，钛质粒的T-DNA区域在细菌细胞中循环。这种反应是由植物细胞分泌的可扩散植物因子(S)诱导的。我们还发现，被认为是抗农杆菌侵染的单子叶植物含有诱导物质(S)，但它不同于双子叶植物。我们尝试从小麦麸皮中提纯植物因子，并确定其分子结构。在纯化过程中，诱导活性被分成两个组分，一个是亲水性的，另一个是疏水性的，这两个组分都是诱导T-DNA环化所必需的。此外，我们还揭示了钛质粒的病毒D位点参与了环化事件。该基因座上的Vird1和D2基因一起是必需的。这些基因编码的产物似乎具有T-DNA边界序列所特有的内切酶或重组酶的活性。农杆菌RMA聚合酶是T-DNA环化的负调控因子，与Vird1和D2的产物竞争。我们试图纯化病毒D蛋白以及其他病毒D蛋白(D3和D4)，并阐明这些蛋白的酶学性质。