MOLBAS Research Group, Department of Plant Molecular Biology, University of Leiden, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands.
EMBO J. 1984 Dec 20;3(13):3043-7. doi: 10.1002/j.1460-2075.1984.tb02255.x.
The host range of Agrobacterium tumefaciens strain LBA649 (pTiAg57) is limited to grapevine and a few other plant species. Its host range was extended through the introduction of the T-region from the wide host range octopine plasmid pTiAch5. In contrast, R prime plasmids harboring the entire wide host range virulence region were unable to achieve this effect. Via site-directed mutagenesis a search was performed to identify the T-DNA genes which were responsible for the observed host range extension. Inactivation of one of the onc-genes (the cyt gene) was found to abolish the capacity of the T-region to extend the host range of LBA649. Therefore, we cloned the cyt gene into a disarmed T-region plant vector and used it in complementation studies with pTiAg57 via the binary vector strategy. We show that the mere presence of the cyt gene from a wide host range Ti plasmid is sufficient to extend the host range of LBA649 to certain plants. We conclude that the limited host range of LBA649 is not caused by a lack of recognition of plants but is mainly due to the absence or inactivity of a cyt gene in the T-region of pTiAg57.
根癌农杆菌 LBA649 菌株(pTiAg57)的宿主范围仅限于葡萄藤和其他几种植物。通过引入来自广宿主八氢番茄红素质粒 pTiAch5 的 T 区,扩展了其宿主范围。相比之下,携带完整广宿主毒性区的 R 质粒不能达到这种效果。通过定点诱变,我们进行了一项搜索,以确定负责观察到的宿主范围扩展的 T-DNA 基因。发现一个致癌基因(细胞基因)的失活会消除 T 区扩展 LBA649 宿主范围的能力。因此,我们将 cyt 基因克隆到一个无武装的 T 区植物载体中,并通过二元载体策略使用它与 pTiAg57 进行互补研究。我们表明,来自广宿主 Ti 质粒的 cyt 基因的存在足以将 LBA649 的宿主范围扩展到某些植物。我们得出结论,LBA649 的有限宿主范围不是由于缺乏对植物的识别,而是主要归因于 pTiAg57 的 T 区中 cyt 基因的缺失或失活。