Development of Efficient, Reproducible and Stable Agrobacterium-Mediated Genetic Transformation of Five Potato Cultivars
Department of Agricultural Genetic Engineering, Faculty of Agricultural Sciences and Technologies, Nigde Omer Halisdemir University, 51240 Nigde, Turkey
Received: 25 December 2018
Accepted: 6 March 2020
potato genetic transformation, genotypes, plant growth regulators, cost-effective protocol
The developments in transformation technology have enabled the scientists to incorporate, mutate or substitute gene(s) leading to a particular trait; advancing it to a point whereonly few technical limitations remain. Genotype dependency and explant types are importantfactors affecting transformation efficiency in potato. In the present study, a rapid, reproducibleand stable Agrobacterium-mediated transformation procedure in potato wasdeveloped by a combination of different plant growth regulators. Leaf discs and internodalexplants of five cultivars of potato, i.e. Lady Olympia, Granola, Agria, Désirée and Innovatorwere infected with Agrobacterium tumefaciens strain LBA4404 containing pBIN19 expressionvector with β-glucuronidase gusA gene under the control of 35S CaMV promoter. Kanamycinwas used as plant selectable marker for screening of primary transformants at concentration of 100 mg/L. Both explants responded positively; internode being more suitable explant forbetter transformation efficiency. Based on GUS histochemical assay, the transformation efficiencywas 22, 20, 18.6, 15 and 10 % using the internodal explant, and 15, 12, 17, 8 and 6 %using leaf discs as explant in Lady Olympia, Granola, Agria, Désirée and Innovator respectively.Furthermore, PCR assays confirmed the presence of gusA and nptII genes in regeneratedplants. The molecular analysis in succeeding progeny showed proper integration andexpression of both genes. The results suggest Lady Olympia as the best cultivar for futuretransformation procedures. Overall, the short duration, rapidity and reproducibility makethis protocol suitable for wider application of transgenic potato plants.