Acta Univ. Agric. Silvic. Mendelianae Brun. 2013, 61(5), 1279-1288 | DOI: 10.11118/actaun201361051279

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

Helena Klenotičová, Iva Smýkalová, Lenka ©vábová, Miroslav Griga
Plant Biotechnology Department, Agritec Plant Research, Zemědělská 16, 787 01 ©umperk, Czech Republic

Optimisation of in vitro regeneration systems of two explant types 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 callogenesis, 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 influenced shoot proliferation, except of variant with ascorbic acid. Citric acid was the best and universal antioxidant in faba bean in vitro cultures and its use is recommended for faba bean genetic transformation experiments.

Keywords: acetosyringone, antioxidants, citric acid, co-cultivation agents, faba bean, organogenesis
Grants and funding:

This study was supported by grants: MSMT No. 1M06030, NAZV No. 91A229.

Received: April 30, 2013; Published: September 28, 2013  Show citation

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Klenotičová, H., Smýkalová, I., ©vábová, L., & Griga, M. (2013). Resolving browning during the establishment of explant cultures in Vicia faba L. for genetic transformation. Acta Universitatis Agriculturae et Silviculturae Mendelianae Brunensis, 61(5), 1279-1288. doi: 10.11118/actaun201361051279
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