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:
Crosstalk (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:
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.
schematic representation (left) and structural model (right) of a transmembrane Receptor Like Kinase
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:
- Genetic analysis: phenotypic and histological analysis of single and double mutants ( modifiers obtained in mutagenesis, TDNA knock out lines)
- 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)
- Identification of a novel interactor genes via differential expression profile of the mutant and wild type early seeds (microarray, transcriptome sequencing), mutagenesis.
- 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.