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Rivista di Biologia Molecolare e Biotecnologie

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Minerva Biotecnologica 2011 December;23(4):93-102

lingua: Inglese

Apical meristems: potent and competent explants for improved miniprep transformation method in tomato

Safdar N.

Department of Biochemistry, Quaid-i-Azam University, Islamabad, Pakistan


Extensive studies have been carried out on optimizing the regeneration and transformation conditions in tomato using different cultivars like Micro-Tom, UC82B, Monalbo, LA2747. Many of these studies reports a laborious protocol with the transformants obtained in a period of no less than three months. This paper reports the comparative transformation events assessed by two protocols in the tomato cultivars Rio Grande and Roma, the most common and practical cultivars grown in Pakistan. Apical meristems proved to be the best and unique explants for obtaining the maximum number of transformants in both the cultivars tested. The miniprep transformation methodology consisted of meristem explants preculturing for two days on cocultivation medium followed by infecting the explants with Agrobacterium tumefaciens strain C58C1 carrying npt II and gus genes for 48 hours and then shifting these explants directly to half strength MS medium containing 40 mg/L kanamycin which resulted in the production of transformants in the shortest time possible of fifteen days. Transformation efficiency achieved in Rio Grande and Roma was 41.3% and 43.5% respectively which establish this miniprep protocol to be competent and genotype independent. Southern blots and analysis of foreign npt II gene inheritance in the next generation indicated the stable T-DNA integration into the plant genome. The developed genetic transformation protocol is simple, efficient and effortlessly applicable in various cultivars of tomato for the transfer of useful agriculturally important genes.

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