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Convergence of developmental pathways: in vitro analysis of organ formation capacity in tomato mutants affecting leaf architecture The in vitro culture of plant cells, tissues and organs, started at the beginning of the 20th century, has been leading to scientific and biotechnological advances ever since. The process of in vitro regeneration depends on the growth conditions as well as the genotype of the explant. Several authors postulated that regeneration can bedivided into steps and that the hormonal balance will determinate the fate of the explant (e.g. formation of roots, shoots or callus). In a previous study, it was found that the mutant Mouse-ears, which exhibits a more complex leaf architecture, has high capacity of shoot regeneration. Hence, in the present work others mutants affecting leaf architecture such as entire, procera, clausa, potato-leaf and Mouse-ears were measured ex vitro and in vitro in the MT background. These assessments aimed to investigate if there are similarities in organogenesis between in vitro and ex vitro. The phenotypic characterization of the genotypes showed that these mutations affect several aspects of plant development and not only the leaf architecture. Many differences observed ex vitro are in part due to the transition from vegetative to reproductive phase. When analyzing the results of in vitro regeneration (formation of adventitious shoots and roots; and callus growing) it is noticed that the organogenetic capacity of each mutant have no correlation with the degree of leaf complexity but with the type of mutation involved. Morphological analysis of in vitro and ex vitro events, such as formation of adventitious shoots and shaping of shoot apical meristem, respectively, denotes similarities and divergences between the processes. The results indicate that probably the mutants entire (constitutive response to auxin due to a loss-of-function in an AUX/IAA) and Mouse-ears (overexpression of a KNOX TKN2) act at the induction step of root and shoot, respectively, in the in vitro regeneration process. On the other hand, procera (constitutive response to gibberellin) acts at the competence step or the arrest of the development after induction. Furthermore, TKN2 showed an important role in the formation of in vitro adventitious shoots. The pathways for ex vitro organ formation (e.g. leaf, root, axillary buds, etc.) are different from that for in vitro. Nevertheless, genes and molecules recruited in the pathway of ex vitro development may interfere in the in vitro organogenesis.

Convergência de vias de desenvolvimento: análise da capacidade de formar órgãos in vitro em mutantes de tomateiro afetando a arquitetura foliar