This study describes the successful formation of floral organ pistil from the callus of pistil explants of Oryza sativa L. For induction of floral organs, different explants--including young embryo, lemma, palea and pistil--were used for callus induction with different combinations of N(6)-benzyladenine and 2,4-dichlorophenoxyacetic acid (2,4-D). High frequencies of callus formation from pistil and young embryo explants were achieved. Floral organs were induced after calli from pistils were transferred to medium containing both zeatin and 2,4-D. The morphological characteristics of the pistil-like organs are very similar to those formed in planta though with minor differences. Further histological study revealed that the in vitro pistil contains an ovule within its ovary. Furthermore, a pistil-specific gene, OsMADS3 used as a molecular marker for pistil identity, was expressed in the pistil-like organs as it was in pistils in the flower of the plant.
The petal is an important floral organ of higher plants. To study the mechanism of petal development, the in vitro regeneration system of petals was established in Petunia. High-frequency induction of petals occurred directly from explants on the media containing the combination of N6-benzyladenine (6-BA) and indole-3-acetic acid (IAA). Expression analysis of genes involved in flower development indicated that these genes were classified into three types. ABERRANT LEAF AND FLOWER (ALF) gene was induced during petal regeneration. Whereas, B-class and E-class genes, and genes involved in cell division were constitutively upregulated. In contrast, C-class and D-class genes were not expressed in explants and regenerated tissues. Further, in situ hybridization analysis showed that both ALF and GREEN PETAL (GP) expression were spatially regulated. The results suggest that differential regulation of gene expression occurs in the presence of hormones during petal regeneration, and hormone-regulated gene expression might be required for petal regeneration. This study provides the preliminary information to understand the mechanism of petal regeneration.
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