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Sugarbeet leaf disc culture: an improved procedure for inducing morphogenesis
Authors:Lowell D Owens  Debra R Eberts
Institution:(1) Plant Molecular Biology Laboratory, Agricultural Research Service, United States Department of Agriculture, 20705 Beltsville, MD, USA;(2) Present address: Environmental Sciences Section, Bureau of Reclamation, United States Department of Interior, Denver Federal Center, 80225 Denver, CO, USA
Abstract:In preparation for gene transfer experiments we investigated factors that might affect the production of shoots and somatic embryos from the wound callus of cultured sugarbeet leaf discs. A complex interaction was found between the leaf disc plating density, the disc culture medium, the source-shoot culture medium and the frequency of disc transfer to fresh medium. The most productive protocol utilized: source shoots maintained on MS medium containing 0.25 mg 1-1 BA; multiple leaf discs (ten 4-mm discs/plate) plated onto an enriched modification of MS medium (RV) containing 1.0 mg 1-1 BA and solidified with 0.3% Gelrite (not permitted to dry during hardening); and transfer of the discs to fresh medium every two weeks during the first month. This standard protocol produced more callus per plate and higher rates of morphogenesis per unit dry weight of callus than did the one-step method of Saunders and Doley. Water availability considerations were found to be critical to obtaining high morphogenic rates. Root induction frequency and quality was superior on shoots transplanted to MS medium containing 1 mg 1-1 NAA as the sole growth regulator compared to IAA at the same concentration.Abbreviations BA N6-benzyladenine - IAA indole-3-acetic acid - NAA agr-naphthaleneacetic acid
Keywords:Beta vulgaris  morphogenesis  regeneration  tissue culture
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