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Research Overview


PhD Project: Receptor kinase - mediated regulation of seed development in A.thaliana


Seed development and formation is a key event in a life cycle of a flowering plant. The proper development of the seed is achieved through coordinated growth of its three components: embryo, endosperm and maternal integuments:

picture4.jpgCrosstalk (black arrows) between different tissues in Arabidopsis seed development: maternal integuments, free nuclear endosperm and embryo (heart stage).



In more important crop species ( wheat, barley, rye, maize) endosperm is an important source of nutrition in human diet:


wheat-kernel.jpg


Coordination between the three key players is mediated by intercellular CELL-CELL COMMUNICATION. This mechanism is thought to involve transmembrane receptor like kinases (RLK) and secreted peptides that act as their ligands.

RLKS

schematic representation (left) and structural model (right) of a transmembrane Receptor Like Kinase



How is the coordination of a seed development coordinated? What are the partners of IKU2 receptor?

EXPERIMENTAL APPROACH

In order to decipher the role of IKU2 receptor in seed development and reveal its interaction with other receptor kinases I have undertaken following genetic, biochemical and transgenic approaches:

  1. Genetic analysis: phenotypic and histological analysis of single and double mutants ( modifiers obtained in mutagenesis, TDNA knock out lines)
  2. Phenotypic analysis of mis - expression transgenic lines ( overexpression:pOp/LhG4 transcription activation system, RNAi knock out lines, 35Spromoter lines), subcellular localisation of the gene driven by its native promoter using GFP and myc tag)
  3. Identification of a novel interactor genes via differential expression profile of the mutant and wild type early seeds (microarray, transcriptome sequencing), mutagenesis.
  4. Biochemical approach: immunolocalisation of the protein, co- immunoprecipitation of the protein complex in order to decipher possible interactors of the protein of interest using functional transgenes driven by the native promoter of the gene of interest.


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