Abstract
SUMMARYThe 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 where only few technical limitations remain. Genotype dependency and explant types are important factors affecting transformation efficiency in potato. In the present study, a rapid, reproducible and stable Agrobacterium-mediated transformation procedure in potato was developed by a combination of different plant growth regulators. Leaf discs and internodal explants of five cultivars of potato, i.e. Lady Olympia, Granola, Agria, Désirée and Innovator were infected with Agrobacterium tumefaciens strain LBA4404 containing pBIN19 expression vector with β-glucuronidase gusA gene under the control of 35S CaMV promoter. Kanamycin was 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 for better transformation efficiency. Based on GUS histochemical assay, the transformation efficiency was 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 regenerated plants. The molecular analysis in succeeding progeny showed proper integration and expression of both genes. The results suggest Lady Olympia as the best cultivar for future transformation procedures. Overall, the short duration, rapidity and reproducibility makes this protocol suitable for wider application of transgenic potato plants.
Highlights
Potato (Solanum tuberosum L.) is a primary non-cereal food crop and ranks 4th in the world because of its high productivity, following corn (Zea mays L.), rice (Oryza sativa L.) and wheat (Triticum aestivum L.) in terms of yield, acreage and value [1]
Leaf disc explants cultured on RSM3 showed good results compared to other two selection media, the response of cultivars varied
The maximum percentage of resistant callus development was observed in Désirée, followed by Lady Olympia, Agria, Granola and Innovator, whereas average number of shoots per explant was high in Désirée and
Summary
Potato (Solanum tuberosum L.) is a primary non-cereal food crop and ranks 4th in the world because of its high productivity, following corn (Zea mays L.), rice (Oryza sativa L.) and wheat (Triticum aestivum L.) in terms of yield, acreage and value [1]. Turkey is an important potato producing country globally. Potato accounts for 3 % of the gross national agricultural product, 3.1 % in 27 EU countries, has a significant impact on Turkish economy [3]. Potato is affected by viral, bacterial and fungal diseases, and nematodes [4]. The crop is affected by pathogens at different stages of growth, from the preharvest to the postharvest [5]. The researchers have incorporated different traits related to biotic and abiotic stress tolerance in potato. These technologies transcend traditional plant breeding methods by allowing the rapid and predictable gene transfer across the species boundaries.
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