Bean tissue culture for introduction of foreign genes

Abstract

The objective of this work was to determine different bean tissue culture parameters necessary to the application of Agrobacterium-mediated transformation technique. Several bean (Phaseolus vulgaris L.) genotypes were cultured in vitro. The protocols for Agrobacterium elimination, after infection of the apical meristems with Agrobacterium tumefaciens and the selective kanamycin concentration were determined. Depending on the genotype, multiple buds were obtained from cotiledonary nodes in a frequency up to 55.7%. The cotiledonary nodes were infected with A. tumefaciens carrying in the Ti plasmid the gene for β-glucuronidase (GUS) under the control of an eucariotic promoter. Regions with GUS activity were detected in 30% of the shoots. The susceptibility of bean to Agrobacterium rhizogenes was also shown. Transgenic roots were obtained after infection of epicotyls with A. rhizogenes