An efficient transformation and VAT (resistance to Aphis gossypii) gene transformation of watermelon (Citrullus lanatus (Thunb.) Matsum and Nakai

In this thesis, optimum media for regeneration and gene transformation technique with Agrobacterium tumefaciens to watermelon (Citrullus lanatus (Thunb.) Matsum and Nakai) plant was studied. The best result obtained from MS medium containing 2 mg/l BAP +0.1 mg/l IAA added, for regeneration of the watermelon seeds which are incubated for germination for 5 days. Regeneration protocol has been achively applied to obtain transformed plants, using Agrobacterium tumefaciens strain C58C1-pmp90 and C58C1-pch32 which is harboring a binary vector pBin19 carring Vat coding region, 2,5 kb of 5? and 3? regulatory sequences.Watermelon seed explants which are treated with bacteria solution cultured in MS media containing 2 mg/l BAP + 0.1 mg/l IAA + 200 mg/l timentin + 7.5 g agar added. To determine the transformant plants, selective media containing (2 mg/l BAP + 0.1 mg/l IAA + 750 mg/l sefuroksim sodium + 200 mg/l kanamisin + 7.5 g agar) was used. The plants which are carried on living in selection medium were transferred to the MSG (medium of shoot growth). After completed their growth, the plants were harvested for DNA isolation. Transformed plants were detected by PCR, Gel Electrophoresis and UV decumantation system. Bands are not seen in the intended region.Key words : Citrullus lanatus, transformation, Agrobacterium tumefaciens, rejeneration, Aphis gossypii