Tepary Bean (<i>Phaseolus acutifolius</i>) <i>Agrobacterium</i> Protocols

Phaseolus beans are among the major legumes for food consumption, especially in Latin America, Africa, and Asia. Tepary bean ( Phaseolus acutifolius L. Gray) is one of the five cultivated species of the genus Phaseolus. This chapter describes an Agrobacterium -mediated transformation protocol for P. acutifolius based on cocultivation of callus, derived from cotyledonary nodes, with Agrobacterium . The selectable marker gene used is neomycin phosphotransferase II ( nptII ), and the selection agent is geneticin. Selection of transgenic callus material is achieved through four to five passages on geneticin-containing medium, after which shoots are induced on medium without selection agent. The protocol as described here has been applied to transform a cultivated variety of P. acutifolius, TB1, and also with some modifications to a wild genotype, NI576 and another cultivated variety, PI440795.