Direct Organogenesis from Cotyledons in Cultivars of Citrus clementina Hort. ex Tan

Abstract

An efficient protocol to induce shoot buds regeneration in Citrus clementina cultivars (“Monreal”, “SRA 63” and “SRA 64”) by direct organogenesis has been developed using cotyledons as explants. Cotyledons transversely cut in three segments and entire ones were cultured on Murashige and Skoog (1962) solidified medium containing vitamins, 500 mg·l−1 malt extract, 50 g·l−1 sucrose and supplemented with three different concentrations of BAP (8.8, 13.2 and 17.6 μM). In all three cultivars the entire cotyledons showed more shoot morphogenic potential than transversely cut ones and after 60 incubation days the optimum BAP concentration was 17.6 μM in “Monreal” (50% ± 2.89% of frequency regeneration) and 13.2 μM in “SRA 63” (33.33% ± 3.33%) and “SRA 64” (25.93% ± 1.85%). In absence of BAP No morphogenesis occurred, demonstrating the absolute requirement of this hormone for shoots induction. The young shoots showed a regular growth in the culture tubes containing the basal medium without hormones, and the rooted plantlets survived after acclimatization. This protocol may find application in Citrus genetic improvement programs