Abstract

PEREZ BERNAL, Maylin et al. Quantitative transient GUS expression in J-104 rice calli through manipulation of in vitro culture conditions.. Rev. colomb. biotecnol [online]. 2009, vol.11, n.2, pp.75-84. ISSN 0123-3475.

This paper purposes suitable conditions for callus induction and co-cultivation with Agrobacterium tumefaciens of J-104 rice cultivar. It was evaluated the effect of different concentrations of 2.4-D and agar, and the inclusion of L-proline and L-glutamine in callus culture medium. The use of 2.5 mg/L 2.4-D and 0.8% agar allowed the highest percentage of embryogenic calli. Callus formation was improved considerably with 500 mg/L of L-proline and L-glutamine in the culture medium. Different factors were studied throughout co-cultivation of calli with A. tumefaciens: inoculation time, co-cultivation temperature, concentration of acetosyringone and co-cultivation period. Transient GUS expression was quantified by fluorometry in all co-cultivated calli. The best results were obtained with the following conditions: 10 min as inoculation time, 100µM acetosyringone in co-cultivation medium, temperature of 20°C, and 3 days as co-cultivation period.

Keywords : Agar; callus; co-cultivation; fluorometric GUS activity.

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