The anatomy and morphology of bud regeneration were investigated in melon (Cucumis melo L.) cv. Galia, which regenerates in vitro only by direct organogenesis from the cotyledon explant. Explants were cut from the cotyledon proximal to the apex from 3-d-old in vitro seedlings, After 3 d on Murashige and Skoog medium with N6-benzyladenine, cell division can be observed in the epidermal layer on the adaxial side in the center of the explant, near the most proximal (wounded) cut edge. Over the next week, the area of the meristem increases laterally. Additional cell layers are added to the meristematic area by cell division in the epidermis. In places the epidermis remains active in cell division. Alongside those active areas there are zones where the epidermis has become inactive, although the subepidermal layers continue to divide. In transverse section, the explant now has small protuberances on the adaxial surface. After 10 d on cytokinin-containing medium, the first signs of development are visible on the adaxial surface adjacent to the proximal cut edge. The protuberances observed after 10 d are neither primordia nor buds, although some meristematic bulges are observed. The first regenerated shoot buds are observed histologically after 15 d, by which time the surface has many protuberances and some small leaves. The first shoot is found by histology after 22 d. By this time the surface is covered with protrusions and leaves, mostly without accompanying buds. The leaves may be produced from the protrusions initially visible after 10 d.
Seedling-derived cotyledon explants of squash (Cucurbita pepo L.) of commercial cultivars True French, Ma'yan and Goldy were regenerated in vitro on Murashige and Skoog medium augmented with 1 mg/l benzyladenine. After 4 weeks in culture small shoots and buds regenerated only on the most proximal cotyledon edge. Culture on an elongation medium with a reduced cytokinin concentration (0.1 mg/l) with or without 1 mg/l gibberellic acid (GA 3 ) facilitated the recovery of shoots. Fresh shoots could be recovered at each subculture of the regenerating mass. Peak productivity was during the third cycle of subculture, and shoot production ceased after the fifth subculture. Culture on elongation medium supplemented with GA 3 was 55% more effective with respect to overall shoot production than that on medium without GA 3 , with 22 shoots recovered in total per explant from the former. Regeneration occurred under both light and dark conditions. All of the shoots tested were diploid. The shoots were rooted and transferred to the greenhouse where they grew and flowered normally.
Hypocotyl explants of three cultivars of melon (Cucumis melo L.) (cvs. Revigal, Topmark and Kirkagac), and a cucumber (C. sativus L. cv. Taoz) rapidly directly regenerated multiple shoots on Murashige and Skoog medium augmented with 4.4 μm benzyladenine. Regeneration from the hypocotyl resulted in nearly 100% diploid shoots, whereas regeneration from the cotyledons resulted in 40% to 70% polyploid regenerants. Regeneration from cotyledon explants of melon cv. Revigal required light, whereas regeneration from hypocotyl explants of melon cv. Revigal occurred in both light and darkness. Direct regeneration also occurred from the hypocotyl of cucumber cv. Taoz in both light and darkness, even though cotyledonary explants did not regenerate buds or shoots under the same conditions. This is the first report of regeneration from the Cucumis genus producing a fully diploid plant population.
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