African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12481

Full Length Research Paper

Development of long-term and reliable in vitro plant regeneration systems for elite malting barley varieties: Optimizing media formulation and explant selection

Gerardo Armando Aguado-Santacruz1*, Ángeles Velázquez-Ordinola1, Blanca Moreno-Gómez1, Luisa Marina Gómez-Torres2, Luís Febronio Díaz-Espino3 and Francisco Paúl Gámez Vázquez4
1Laboratory of Plant Molecular Physiology, Campo Experimental Bajío, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Km 6.5 Carretera Celaya-San Miguel de Allende, Celaya, Guanajuato 38110, México. 2Departamento de Ingeniería Química y Ambiental. Universidad Nacional de Colombia,Avenida Carretera 30 No. 45-03, Bogotá, D.C., Colombia. 3Programa de Uso Eficiente de Agua, Campo Experimental Bajío, Instituto Nacional de Investigaciones Forestales y Agropecuarias, Km 6.5 Carretera Celaya-San Miguel de Allende, Celaya, Guanajuato 38110, México. 4Fundación Guanajuato Produce, Calle Nuevo León 603, Colonia Alameda, Celaya, Guanajuato 38050, México.
Email: [email protected], [email protected]

  •  Accepted: 07 December 2011
  •  Published: 26 December 2011

Abstract

The response to in vitro tissue culture of five important Mexican malting barley cultivars namely ‘Armida’, ‘Esmeralda’, ‘Adabella’, ‘Esperanza’ and ‘Alina’, was evaluated. Callus induction and plant regeneration were evaluated in shoot apices and immature barley embryos harvested eight, 12, 16, 20 and 24 days after pollination. These explants were cultured on 22 media formulations that included Murashige and Skoog (MS) and Chu (N6) media amended with two carbon sources and different concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D), 3,6-dichloro-2-methoxybenzoic acid (Dicamba), 6-benzyladenine (BA), casein hydrolyzate and L-proline. No regenerable callus lines could be recovered from the experiments using shoot apices as the starting material. Conversely, immature embryos harvested at 12 to 16 days after pollination (1.0 to 1.5 mm long) and cultured on formulations containing the basal MS medium supplemented with 2 mg L-1 of 2,4-D or Dicamba resulted in the production of nodular and friable embryogenic calli with regenerative capacity in all barley cultivars. A medium containing MS, Dicamba and maltose generated the highest percentage of embryogenic calli formation (2.1 in ‘Adabella’ vs. 23.4 in ‘Alina’ cultivar) with the maximal regenerative potential (7.2 in ‘Armida’ vs. 9.1 recovered plants per gram FW callus in ‘Esperanza’ cultivar). Using this medium, ‘Esmeralda’ embryogenic callus lines have been maintained for more than five years without loss of regenerability or the capacity for producing normal plants.

 

Key words: Barley, somatic embryogenesis, plant regeneration, immature embryos.

Abbreviation

2,4-D, 2,4-Dichlorophenoxyacetic acid; Dicamba, 3,6-dichloro-2-methoxybenzoic acid; Picloram, 4-amino-3,5,6-trichloropicolinic acid; BA, 6-benzyladenine, MS, Murashige and Skoog medium; N6, Chu medium; IAA, indole acetic acid