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首页> 外文期刊>Cytology and genetics >Obtaining Transgenic Potato Plants Expressing the Human Lactoferrin Gene and Analysis of Their Resistance to Phytopathogens
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Obtaining Transgenic Potato Plants Expressing the Human Lactoferrin Gene and Analysis of Their Resistance to Phytopathogens

机译：获得表达人乳铁蛋白基因的转基因马铃薯植物并分析它们对植物病变的抗性

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摘要

The human lactoferrin gene was transferred into genomes of several potato (Solanum tuberosum) cultivars of Ukrainian breeding using the Agrobacterium-mediated transformation. The plasmid vector pBIN35LF carrying the human lactoferrin gene hLf controlled by the 35S cauliflower mosaic virus promoter (CaMV35S) and the octopine synthase terminator, as well as the selective marker neomycin phosphotransferase II gene (nptII) conferring the resistance to kanamycin, was used. As a result of selection, 44 lines of Vernisage, 26 lines of cv. Levada, 25 lines of cv. Svitanok Kyivskyi, and 16 lines of cv. Zarevo cultivars resistant to 100 mg/L of kanamycin were obtained. PCR and Western blot analyses were carried out for transformed lines with primers specific to the hLf gene and a monoclonal antibody against lactoferrin to confirm the transgenic nature of selected tomato plants and hLf gene expression. The selected transgenic potato lines were tested for resistance to bacterial and fungal phytopathogens. The disk diffusion assay revealed that the juice of transgenic potato lines possesses an antibacterial effect against phytopathogenic bacteria Ralstonia solanacearum (causing potato brown rot) and Clavibacter michiganensis subsp. sepedonicus (causing potato ring rot). The resistance of transgenic potato plants to late blight was investigated by in vitro infection of plants with the Phytophthora infestans isolate. As a result, it was found that the obtained transgenic potato lines have enhanced resistance to P. infestans as compared to the control. Thus, the obtained data show that the transfer of the hLf gene into the potato genome enhances potato's resistance to bacterial and fungal pathogens.
机译：使用农杆菌介导的转化转移到几种马铃薯（Salanum Tuberosum）品种的几种薯类（Solanum Tuberosum）品种的基因组中。使用由35S花椰菜马赛克病毒启动子（CAMV35S）和章鱼合酶终止剂控制的人乳铁蛋白基因HLF的质粒载体PBIN35LF以及赋予赋予卡那霉素的耐药性的选择性标记新霉素磷酸转移酶II基因（NPTII）。由于选择，44行葡萄酒，26行CV。 Levada，25行CV。 Svitanok Kyivskyi和16行CV。获得抗抗100mg / L的卡那霉素的Zarevo品种。对具有特异于HLF基因的引物和针对乳铁蛋白的单克隆抗体的转化线进行PCR和Western印迹分析，以确认所选番茄植物和HLF基因表达的转基因性质。测试所选择的转基因斑点系对细菌和真菌植物病变的抗性。磁盘扩散测定显示转基因薯系的果汁对植物疗法细菌的抗菌作用具有抗菌作用，对植物疗法细菌茄科（导致马铃薯棕色腐）和克拉维奇密歇根群。术术（导致马铃薯环腐烂）。通过对植物的体外感染植物的植物与植物植物嗜患者分离物进行体外感染，研究了转基因马铃薯植物的抗性。结果，结果发现，与对照相比，所获得的转基因薯系具有增强对P.缺血性的抗性。因此，所获得的数据表明，将HLF基因转移到马铃薯基因组中提高了马铃薯的细菌和真菌病原体的抵抗力。

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