Agrobacterium mediated transformation of cucumis melo with replicase gene from papaya ringspot virus and regeneration of transformed paints

Abstract

Transgenic muskmelon (Cucumis melo) plants were produced efficiently by inoculating cotyledon explants with the Agrobacterium tumefaciens strain EHA 101 bearing a Ti binary vector. The T-DNA region of this vector includes chimeric genes for neomycin phosphotransferase II (NPTII) and (3-Glucuronidase (GUS). Additionally the plant expressible replicase gene (Nib) of papaya ringspot virus (PRV) was flanked by the NPT II and GUS genes. After co-cultivation for three days, explants were transferred to muskmelon regeneration medium with kanamycin for the selection of transformed tissue. Shoot regeneration occurred within 3-5 weeks after culturing. Detection of GUS activity in 5 plant lines verified the transformation of the T-DNA unit and two of them were further analyzed by PCR amplification for the presence of the replicase gene. These results show that the Agrobacterium mediated gene transfer method and regeneration via tissue culture are effective methods for the transfer of foreign genes into Cucumis melo.