Plant Cell Culture Protocols

Rosa Maria Escobedo-Gracia-Medrano, Carlos Iván Cruz-Cárdenas, Lucila Aurelia Sánchez-Cach, José Roberto Ku-Cauich, Wilma Aracely González-Kantún, Escobedo-Gracia-Medrano, Rosa Maria, Cruz-Cárdenas, Carlos Iván, Sánchez-Cach, Lucila Aurelia, Ku-Cauich, José Roberto, González-Kantún, Wilma Aracely

Most cultivated bananas (Musa spp.) are polyploids, and their fruits are seedless and propagated exclusively vegetatively; however, they can also be cloned by micropropagation techniques, viz., direct organogenesis (DO) or somatic embryogenesis (SE). Banana indirect SE (ISE), with an embryogenic callus phase, is possible using young male or female flowers as direct explant depending on the genotype or shoot tips (scalps). For the False Horn Plantain, cv. Curraré (AAB, plantain subgroup), which has a degenerating male bud, female flowers are used to regenerate plants through ISE. Here, a protocol for increasing the number of initial explant material from a single mother plant and its embryogenic response is described. For those purposes, hands with young female buds are in vitro proliferated in the presence of 1 μM indole-3-acetic acid and 2.5 μM thidiazuron. Friable embryogenic cultures, here called ISE-2, obtained from the new proliferative secondary female bud clusters are initiated on medium containing auxins. Embryogenic suspensions are then established from the ISE-2 cultures. Regeneration of plants is achieved from embryogenic suspensions after plating on semisolid medium free of plant growth regulators; greenhouse acclimatized plantlets are ready for banana farming. This study demonstrates that proliferative female buds are a proper choice for ISE.