A reliable system for the transformation of cantaloupe charentais melon (Cucumis melo L. var. cantalupensis) leading to a majority of diploid regenerants

An efficient system of transformation leading to a majority of transformed diploid plants from led explants of Cucumis melo L. var. Cantalupensis (cv. Vedrantais) was developed. Several regeneration protocols using cotyledon or leaf explants were analysed with particular emphasis on the regeneration efficiency and the ploidy level of the regenerated melon plants. The use of leaf explants excised from 10 day-old seedlings, cultured in Murashige and Skoog's medium supplemented with 1 mu M 6-benzylaminopurine (BAP) and 1 mu M 6-(gamma,gamma-dimethylallylamino)-purine (2iP), resulted in a high regeneration frequency (73%). In these conditions, more than 84% of the regenerated plants were found to be diploid. Addition of an Agrobacterium-mediated transformation step did not significantly change the percentage (81.8%) of diploid plants regenerated. This protocol was successfully used to produce diploid transgenic melon plants expressing the antisense ACC oxidase gene, encoding ACC oxidase which catalyses the last step of ethylene biosynthesis. Ethylene production and ACC oxidase activity of the leaf explants from transgenic plants was reduced by more than 80% as compared to the control untransformed tissues. This transformation/regeneration method could be routinely used for the introduction of other genes of interest in melon. (C) 2000 Elsevier Science B.V. All rights reserved.