Investigating the role of DA1 in growth control

Increasing global demand for food is a major issue facing modern day agriculture. For crops such as wheat and rice, where the seed constitutes the harvestable yield, the engineering of larger seeds provides a possible strategy for yield improvement. A detailed understanding of the growth of plant or...

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Bibliographic Details
Main Author: Dumenil, Jack
Published: University of East Anglia 2013
Subjects:
570
Online Access:http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.614541
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Summary:Increasing global demand for food is a major issue facing modern day agriculture. For crops such as wheat and rice, where the seed constitutes the harvestable yield, the engineering of larger seeds provides a possible strategy for yield improvement. A detailed understanding of the growth of plant organs in general is paramount if such advances are to be made. Utilising previously characterised regulators of plant organ growth, this thesis explores the molecular mechanisms involved in the setting of final organ size. This thesis capitalises on previous studies that have identified DA1 as a negative regulator of organ growth; it explores the role of the DA1 protein and investigates its interactions with other proteins. In vitro studies reveal that DA1 forms homo-­‐ and hetero-­‐multimeric complexes with its sister protein DAR1 and in vitro and in yeast assays reveal interactions between DA1 and the transcription factor TCP15 and the growth-­‐regulating receptor-­‐like kinase TMK4. In addition, biochemical assays described in this thesis identify an active ubiquitin interacting motif (UIM) in the N-­‐terminal region of DA1 and an ubiquitin-­‐activated metallopeptidase in its C-­‐terminal region. Further studies reveal that, in addition to being activated by the RING E3 ligases EOD1/BB and DA2, the DA1 peptidase is active towards both EOD1/BB and DA2. In vitro and in vivo studies demonstrate that DA1 cleaves a peptide fragment from the N-­‐terminus of EOD1 and the C-­‐terminus of DA2. Finally, this thesis reports two genetic screens carried out in two separate Arabidopsis mapping populations in order to identify novel regulators of organ growth. Analyses of petal and seed phenotypes in the MAGIC RIL-­type population and in a natural Swedish population identify novel and a priori candidate genes for further characterisation.