New insights into the in vitro organogenesis process: the case of Passiflora

We present evidences that ultrastructural electron microscope findings are valuable ways to understand the in vitro regeneration process, in particular in the yellow passion fruit. Shoot-regeneration was induced in hypocotyl and leaf-derived explants using 4.44 mu M BAP, and the entire organogenic process was analyzed using conventional histology, scanning and transmission electronic microscopy. Both direct and indirect regeneration modes were observed in hypocotyl explants, but only direct regeneration occurred in leaf-derived cultures. In the direct pathway from both explant types, meristemoids developed into globular structures, here called protuberances. The peripheral meristematic layers of the protuberances displayed ultrastructural characteristics indicative of a high metabolic activity, and only these cells originated shoots and leaf primordia, the latter being frequent when leaf explants were used. Moreover, the peripheral cells of the protuberances derived from leaf explants lost adhesion during the culture, diminishing the regeneration rates. We recommend the use of hypocotyls as a source of explant to obtain shoots as well as a genetic transformation system for the yellow passion fruit. However, the direct pathway is preferred because a type of amitosis occurred in the peripheral cells of hypocotyl-derived calli, which has the potential to result in genetic instability of the regenerating plants/tissue.