Biotecnología Vegetal (Jul 2017)

Effect of parameters involved in the transformation via <i>Agrobacterium tumefaciens</i> in the regeneration of <i>Phaseolus vulgaris</i> L. cultivars 'ICA Pijao' and 'BAT 93'

  • Dianella Iglesias,
  • Lyselle Ruiz de León,
  • Damaris Torres,
  • Raúl Collado

Journal volume & issue
Vol. 17, no. 3

Abstract

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Phaseolus vulgaris is a crop widely used in human consumption. Its production is affected by the incidence of biotic and abiotic factors. Genetic transformation provides the possibility of reducing these affections, but the low efficiency in vitro regeneration constitutes a limiting factor for the development of protocols for obtaining plants transformed in this species. The aim of this work was to determine the effect of bacterial suspension concentration, vacuum infiltration time and co-culture on the regeneration of P. vulgaris shoots in 'ICA Pijao' and 'BAT 93' cultivars. Green nodular calli were inoculated with a suspension of A. tumefaciens at a DO600 0.1, 0.2 and 0.3. Three times of infiltration by vacuum were evaluated: 1, 2 and 3 min and co-culture period of three and five days. The concentration of the bacterial suspension, the vacuum infiltration time and the co-culture period affected the regeneration of shoots of P. vulgaris cv. 'ICA Pijao' and 'BAT 93'. The highest percentages of P. vulgaris shoot regeneration were achieved by inoculating the calli with a bacterial suspension adjusted at DO600=0.1, without vacuum infiltration and 3 days in co-culture. The results of this research constitute the first contributions to the scientific community in show the effect of vacuum infiltration on the regeneration of P. vulgaris tissue inoculated with A. tumefaciens. Keywords:co-culture, vacuum infiltration, regeneration, bacterial suspension