Isolation and Analysis of New Molecular Markers for Early Embryogenesis in Rice(Oryza sativa L.)

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  • Isolation and Analysis of New Molecular Markers for Early Embryogenesis in Rice (<i>Oryza sativa</i> L.)
  • Isolation and Analysis of New Molecular

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Two cDNA fragments that preferentially accumulate in very young rice embryos at 4 days after anthesis (DAA) were isolated using simplified differential display. The first morphological change is observed in the embryos at 4 DAA and most of the embryo tissue has already differentiated by 7 DAA. RT-PCR was used to quantitate the expression levels of the genes corresponding to the two cDNA clones, REE5 and REE8. They were expressed at higher levels in the embryo at 4 DAA than in the embryo at 7 and 14 DAA as well as in the pistil 1 day before anthesis. The consistent result was obtained by in situ hybridization experiments using REE5 antisense probe. REE5 gene was expressed in the whole embryo at 4 DAA and 5 DAA. The lower expression levels were observed in the embryos at 3 and 7 DAA. RT-PCR analysis revealed that both the REE5 and REE8 transcripts also accumulated in inflorescence meristems prior to the emergence of floral organs, but not in roots, leaves or flowers. The common feature of the young embryos and the inflorescence meristems is the onset of differentiation of various tissues. These may be useful molecular markers for analysis of the events of differentiation during early embryogenesis.

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