PJB-2017-845
Transformation and molecular analysis of tomato transformed with drought resistant ATAF1 gene
Muhammad Awais
Abstract
Tomato (Solanum lycopersicum L.) is one of the important vegetable crops grown worldwide. It not only provides basic nutritional supplements to the human body but also provides different health benefits. However, its production at large scale is hampered by many limiting factors such as biotic and abiotic stresses. Among the different abiotic stresses, drought poses drastic impact on tomato yield. Drought stress is controlled by many transcription factors that not only regulate the stress responsive mechanism but also facilitate the growth and development of plants. NAC is an important stress related transcription factor genes family, and the ATAF1 gene, a member of NAC family is involved in ABA signaling and stress response. This study focuses to genetically transform tomato by inserting drought resistant ATAF1 gene via Agrobacterium mediated transformation. For this purpose, the ATAF1 gene was cloned in the pK7WFG2 vector having kanamycin selectable marker and then it was introduced in the Agrobacterium tumefaciens strain GV3101 through heat shock method. The tomato cotyledon and hypocotyl ex-plants of variety “Riogrande” were cultured on callus induction medium (MS + 2.5 mg/l IAA + 2 mg/l BAP). The calli were infected with the transformed Agrobacterium tumefaciens strain GV3101 containing ATAF1 gene and selection was carried out on the kanamycin selectable medium (100 mg/l Kan), and regenerate on MS medium containing 1 mg/l IAA + 1 mg/l BAP. Out of 216 putative transformed calli, 13 calli were able to regenerate on the selection medium. Out of the 13 calli, three transgenic tomato plantlets, genetically transformed by ATAF1 gene were recovered, and these were further confirmed through PCR analysis and are under study for stress related experiments.
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