Buziashvili A. Obtaining of genetically modified plants of Solanaceae family with the human lactoferrin gene to enhance their resistance to phytopathogens

The study presents the opportunity of the use of genetic transformation of important crops, tomato and potato, with human lactoferrin gene (hLf) to increase their resistance to bacterial (Ralstonia solanacearum, Clavibacter michiganensis) and fungal (Phytophthora infestans, Fusarium sambucinum) phytopathogens. Transformation of tomato cultivars Money Maker and Lahidny and potato cultivars Vernisage, Svitanok Kyivskyi, Levada and Zarevo was carried out with the use of Agrobacterium-mediated method. For that A. tumefaciens strain EHA105 carrying pBin35Lf plasmid vector containing hLf gene under control of 35S promotor of cauliflower mosaic virus (CaMV35S), and also nptII gene conferring resistance to kanamycin was used. Selection of transgenic tomato and potato lines was carried out on the medium supplemented with 100 mg/l kanamycin. Integration of hLf gene was confirmed by PCR analysis with primers specific to this gene. Expression of lac-toferrin protein in transgenic lines was confirmed using Western blot analysis with the use of specific monoclonal antibodies against lactoferrin. The inhibition of bacterial and fungal growth after treatment with the samples from transgenic plants was shown with the use of agar diffusion assay. With the use of inoculation of transgenic tomato and potato plants with conidia of P. infestans, potato plants – with conidia of F. sambucinum the increase of the resistance of transgenic lines to late blight was established from 1 to 7 points, and to dry rot – from 2 to 7 points of 9-point scale. The results of the present work show the advantages of the genetic transformation of the valuable tomato and potato cultivars with human lactoferrin gene to enhance their resistance to bacterial and fungal phytopathogens