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Functional analysis of two cell cycle regulators required for male-germ line development in Arabidopsis

Reference	BB/C004205/1
Principal Investigator / Supervisor	Professor David Twell
Co-Investigators / Co-Supervisors
Institution	University of Leicester
Department	Biology
Funding type	Research
Value (£)	254,523
Status	Completed
Type	Research Grant
Start date	01/12/2005
End date	30/11/2008
Duration	36 months

Abstract

The DUO genes encode two proteins with essential roles in the control of male germ line development. Both DUO genes are required for progression of the generative cell cycle from S to M phase. Whereas DUO1 mRNA appears to be expressed specifically in developing spores, DUO2 transcripts are present in both male gametophytic and sporophytic tissues. The primary objective is to elucidate the gametophytic role of DUO1 and DUO2 to extend our knowledge of the molecular networks involved in cell cycle progression during gametogenesis. The specific aims of the work programme are: 1. To analyse the expression of DUO1 and DUO2 genes and cellular localisation of the DUO1 and DUO2 proteins. Promoter-GUS fusion analysis will be used to monitor DUO1 and DUO2 expression in gametophytic and sporophytic cell types. Non-invasive fluorescent protein imaging and immunolocalisation will be used to monitor the dynamics of DUO protein localisation. 2. To identify and characterise DUO1 and DUO2-interacting proteins. To identify DUO interacting proteins we will use yeast two-hybrid screening. The role of positive DUO interacting proteins in male germ line development will be tested by phenotypic analysis of cell cycle progression in insertional mutants. 3. To investigate changes in phenotype and gene expression resulting from the ectopic expression of DUO proteins. This approach will use microarray hybridisation to identify changes in target gene expression resulting from the ectopic expression of DUO proteins. Putative target genes regulated by DUO will be investigated by insertional mutagenesis and phenotypic analysis. Overall, the results will contribute significantly to understanding the mechanisms of male gametogenesis and cell cycle control and will be of future value in understanding the evolution of heterochrony in gametophytic development. The information may also be useful for the targeted manipulation of gametogenesis, ploidy level and could also provide tools for the manipulation of gene flow in transgenic crops.

Summary

unavailable

Committee	Closed Committee - Genes & Developmental Biology (GDB)
Research Topics	Microbiology, Plant Science
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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