In vitro cloning of date palm phoenix dactylifera l., cv. Deglet bey by using embryogenic suspension and temporary immersion bioreactor (tib)

Abstract

The present work is the first report on in vitro regeneration of an elite date palm cultivar, Deglet Bey (Mnakher) through both somatic embryogenesis and direct shootformation from young leaf explants cultured on MS agar-solidified medium supplemented with 10 mg.l−1 2,4-dichlorophenoxyacetic acidfor 8 months. Factors affecting embryogenic callus and shoot initiation, including antioxidants, adsorbents and 2,4-D concentrations, were investigated. Embryogenic suspensions and culture of shoots in temporary immersion bioreactor (TIB) were developed to improve differentiation of embryogenic callus and proliferation of regenerated shoots, respectively. The yield of cotyledonary somatic embryos produced in half-strength MS liquid media, especially when enriched with 2 mg.l−1 2,4-dichlorophenoxyacetic acid, were shown to be greater than in agar-solidified medium, namely 501 versus 29 per 0.5 g fresh weight of embryogenic callus. The culture of shoot clusters in TIB with an immersion frequency of 5 min every 8 h with MS liquid medium containing 0.04 mg.l−1 α-naphthalenacetic acid, 0.2 mg.l−1 6-benzylaminopurine 0.02 mg.l−1 kinetin for 6 weeks has clearly improved the yield of regenerated shoots per 15 g of shoot clusters. Indeed, it increased 5.5-fold in comparison with that regenerated on agar-solidified medium. For development into plantlets, cotyledonary somatic embryos and regenerated shoots were cultured on MS agar-solidified medium free of 2,4-D and MS medium comprising 0.1 mg.l−1 NAA, respectively. © 2009 Taylor and Francis Group, LLC.