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Molecular control of ovule development and early embryogenesis

Objectif



Plant breeding and seed production relies on the sexual reproduction process. This process begins in the ovule with the fertilization of an egg cell by a sperm cell to form a zygote and results in the formation of a seed containing a multi-celled bipolar embryo in which the fundamental body organization of the plant has been established. Despite the extensive knowledge of morphogenetic aspects and the availability of a variety of mutants, little is known about the molecular mechanisms governing ovule and embryo development in plants. The current proposal aims at the isolation and characterization of regulatory genes involved in this developmental pathway and the elucidation of their function by a loss- or gain of function mutagenesis approach. The dicot model species petunia and worlds most important cereals (rice, wheat and maize) will be studied to ensure an efficient extrapolation of basic knowledge to applied plant biotechnology for economically important crops. Each plant system in this integrated project will contribute its unique technical advantage and specific plant materials to study the various developmental processes in ovule formation and the early stages of zygotic embryogenesis. Highly specific cDNA libraries are already available from a number of stages as sources for gene cloning and a couple of interesting genes have already been isolated. These provide an excellent starting point for the current proposal .
The strategy involves the isolation of regulatory genes and genes that are expressed during specific stages of ovule and embryo development. More specifically, genes coding for MADS box and Homeo box transcription factors and their direct target genes will be isolated. These transcription factors are known to act as molecular switches in the determination of cell fate and identity. Protein kinases are part of molecular signalling pathways which lead to the induction of developmental processes. The role in ovule development and induction of embryogenesis as well as their interaction with the transcription factors will be examined. Genes involved in meiosis, the formation of the embryo sac, egg cell and zygote will be isolated by means of differential screening approaches and yeast mutant complementation experiments. The exact role of the isolated genes in ovule and embryo development will be determined by a PCR-based 'target selected gene inactivation' using populations of highly mutagenic petunia and maize transposon lines. In addition, gene expression will be modulated by antisense/cosuppression and ectopic expression studies in all plant species involved in this project. These approaches will be completed by a very careful analysis of the obtained mutants at molecular, cellular and morphological level, leading to an as complete as possible description of the function of the isolated genes.
All these complementary approaches will substantially increase our knowledge of the developmental programmes governing the formation of the zygotic embryo. In this way, we also might obtain important clues on the control of apomixis. Furthermore, this project will deliver a large number of well-defined novel regulatory genes of potential importance for agriculture and plant biotechnology. Highly-specific promoters will be exploited for the introduction of inducible female sterility in crop plants. Other genes and constructs might be applicable to improve quality aspects of embryo and seed or to control sexual and/or apomictic reproduction processes. Potential valuable traits for biotechnology, plant breeders and ultimately farmers will be tested by the industrial partners in the project.

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Coordinateur

Dienst Landbouwkundig Onderzoek
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Adresse
1,Droevendaalsesteeg
6700 AA Wageningen
Pays-Bas

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