Abstract
A protocol for Agrobacterium-mediated transformation was developed for tomato plant (cv. Summer set). Transformation was carried out using disarmed A. tumefaciens strain LBA4404 harbouring a binary vector pITB-AFP. Factors that affect transformation/regeneration protocols were optimized in a series of experiments. Results indicated that exposure of cotyledonary explants to Agrobacterium inoculums of 0.8 O.D600 for 30 mins, selection on Hygromycin-containing medium (25mg hygromycin/L) and subsequent regeneration on MS medium supplemented with 2.5mg/L BA and 1.0 mg/L IAA resulted in transformation efficiency of 7%. GUS expression was observed in transformed tomato shoots but never in the control plants. PCR amplification of DNA extracted from the transformed tissues demonstrated the generation of the expected amplicon, corresponding to AFP gene. This result strongly verifies the successful transformation of the tomato cultivar Summer set, an endeavour which is reported for the first time in Sudan. Moreover, this protocol paves the way for problem solving-applications encompassing other Sudanese crops of economic importance.
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