Factors affecting protoplast culture of Cucumis melo 'Green Delica'

Abstract

Hypocotyls, cotyledons and etiolated half-expanded leaves of Cucumis melo 'Green Delica' were used as explants for protoplast isolation and culture. Protoplasts isolated from cotyledons and etiolated half-expanded leaves cultured in Durand, Potrykus and Donn (DPD) medium supplemented with 0.9 μM benzylaminopurine (BAP), 3.6 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1% sucrose, using the agarose bead culture method, were able to form cell walls and subsequently go through cell division. Pretreatment of half-expanded leaf explants in the dark for 14 d provided the best material for protoplast isolation and cell division. Approximately one third of protoplasts from etiolated half-expanded leaves formed microcolonies. For hypocotyl protoplasts, none of the treatments used were suitable to induce cell division. There was no significant difference between sucrose, glucose, and sucrose plus glucose, in culture media on the plating efficiency of leaf protoplasts of C. melo 'Green Delica'; however, bigger colonies were formed in media supplemented with 1% sucrose. No shoot or whole plant regeneration was achieved. However, the methods reported here provide further information on C. melo protoplast culture.