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Revista de Biología Tropical
On-line version ISSN 0034-7744Print version ISSN 0034-7744
Abstract
SOLIS-RAMOS, Laura Yesenia et al. Agrobacterium tumefaciens-mediated transformation of common bean (Phaseolus vulgaris) var. Brunca. Rev. biol. trop [online]. 2019, vol.67, n.2, pp.83-94. ISSN 0034-7744. http://dx.doi.org/10.15517/rbt.v67i2supl.37208.
Common bean is a crop recalcitrant to in vitro regeneration and therefore it lacks an efficient transformation protocol that can be reproduced using A. tumefaciens. The main goal of this study was to establish a protocol for A. tumefaciens mediated transformation of Phaseolus vulgaris var. Brunca by marker genes (gusA and nptII) together with the gene for trehalose-6-phosphate synthase from Saccharomyces cerevisiae (TPS1) used in other species to increase tolerance to abiotic stress. The β-glucuronidase activity was detected in 45 % of the LBA4404 ElectroMAX® pCAMBIA1301 infected explants. Transformed explants regenerated new shoots after four to five months period in a kanamycin rich media. Surviving plants were evaluated by PCR and presented an 0.5 % efficiency of transformation. The established protocol for genetic transformation of common bean has two additional advantages with respect to previous reports: (1) it allows for obtaining transformed regenerants and (2) the genetic transformation was stable for the selective gene.
Keywords : common bean; recalcitrant species; genetic transformation; A. tumefaciens; gusA activit; nptII.