Abstract
An efficient protocol for Agrobacterium-mediated transformation of tomato cultivars Sandra and Rocky was conducted to examine the possibility of producing transgenic tomato plants cultivars harbouring the nptII gene, conferring kanamycin resistance. To achieve this aim, tomato cotyledon explants were transformed using EHA105 Agrobacterium tumefaciens strain harboring the binary vectors pBI121 which contains Gus gene, and neomycin phosphotransferase II (nptII) as selectable marker gene under the control of a CaMV35S promoter and nopaline synthase (nos) Terminator. Transformant detection was carried out in three distinct ways. First antibiotic selection, Kanamycin at a concentration of100 mgl-1 found to be efficient for this purpose. Second histochemical GUS assay revealed the presence of blue colored zones in a number of shoots and leaves for both in vitro and the greenhouse-grown transgenic plants. Third PCR analysis indicated positive result by showing the fragment for nptII gene in tested transformants, while was absent in non-transgenic control (wild type). On the other hand, the results showed that Sandra cultivar was more efficient for regeneration and subsequently transformation frequency than Rocky cultivar, which record 26.66% of transformation frequency compared with 11.57% in Rocky cultivar.
Highlights
Transformation systems refers to the capacity for introducing foreign genes into plants
Plant transformation is an important genetic engineering tool for introducing foreign genes into the genomes of plant species and refers to the transfer and incorporation of engineered genes or plasmids into the plant genome to generate genetically modified plants known as transgenic plants (7)
Plant transformation is a core research tool in plant biotechnology, it is a practical tool for transgenic plant development that is done by several techniques for transfer of isolated genetic materials into a viable host cell. (17)
Summary
Transformation systems refers to the capacity for introducing foreign genes into plants. Plant transformation is an important genetic engineering tool for introducing foreign genes into the genomes of plant species and refers to the transfer and incorporation of engineered genes or plasmids into the plant genome to generate genetically modified plants known as transgenic plants (7). It is an essential biotechnological approach for indepth research of plant growth, and a promising tool for modern agriculture, for the introduction and selection of desired crop traits. It confers transgenic plants resistance to the antibiotic, kanamycin. (3)
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