Resolving browning during the establishment of explant cultures in Vicia faba L. for genetic transformation

Optimisation of two in vitro regeneration systems for low-tannine cultivars of faba bean based on culturing of shoot apices and cotyledonary nodes were provided by usage of various antioxidants - ascorbic acid, citric acid, glutathione and activated charcoal. In subsequent testing, the combined effects of antioxidants with transformation co-cultivation compounds acetosyringone and L-cysteine was studied. The application of antioxidants lead to decreased caullogenesis, citric acids treatments (50 mg.l-1) dramatically decreased necrotic response of explants. However, citric acid, used together with ascorbic acid completely inhibited shoot growth in shoot apex cultures. Glutathion evoked hyperhydricity of explants. Activated charcoal induced rooting on media which are commonly used for shoot proliferation. Combination of acetosyringone with antioxidants did not negatively influenced shoot proliferation, except of variant with ascorbic acid. Citric acid worked as the best and universal antioxidant in faba bean in vitro cultures utilizable and its use is recommended for faba bean genetic transformation experiments.

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