BBSRC Thematic Group: Frontier Bioscience
Plasmodesmata are membrane-lined cytoplasmic channels which are of fundamental importance to plant development, stress responses and defence against pathogens. They also serve as conduits for infection by plant viruses, major crop pathogens with a global agro-economic impact. Building on recent advances in understanding plasmodesmata architecture and virus transport by the Tilsner lab, and a novel method for identifying protein-protein interactions developed by the Hemsley lab, this project will characterize a contractile machinery at plasmodesmata putatively required for the regulation of intercellular communication in plants, and likely co-opted for virus transport.
Several actin-associated proteins have been localized to plasmodesmata and in some cases shown to be involved in regulation of plant cell-cell connectivity and virus transport. The aims of the project are:
• Analyse interactions between actin-related proteins at plasmodesmata
• Identify additional components of the putative contractile structure
• Test effects of individual and combined knock-out mutations on plant development, cell-cell connectivity, virus transport and plasmodesmata ultrastructure.
The PhD candidate will use a novel system for detection of protein-protein interactions in vivo to identify new components of a putative actin-related contractile system at plasmodesmata. The plasmodesmal localization of new candidates will be confirmed by fluorescence and electron microscopy. Co-immunoprecipitation and yeast-two-hybrid assays will be used to analyse reciprocal interactions between all previously known and newly identified components. Functional relations between these proteins will be probed using crosses of Arabidopsis knock out lines, which will be analysed for developmental phenotypes, virus susceptibility and intercellular molecular transport. The student will travel to a collaborating lab for ultrastructural (electron tomography) analysis of plasmodesmata morphology in mutant lines.
The project will provide in-depth training in state-of-the-art technologies such as: restriction-free cloning (Gibson assembly and Gateway); protein biochemistry and mass spectrometry; advanced light (confocal, FRAP, SIM super-resolution) and electron microscopy; transient and stable plant transformation; handling of genetically modified crop pathogens in class 2 containment laboratories and glass houses.
The University of St Andrews is the oldest university in Scotland and consistently ranked as amongst the UK’s top ten universities, currently ranked 2nd place in the Guardian University Guide 2020. The University of Dundee is one of Scotland’s leading universities, ranked 4th for student satisfaction in the 2019 UK National Student Survey. The Dundee Plant Science department is based at the James Hutton Institute, a world-leading plant research center with particular focus on molecular plant-pathogen interactions which offers comprehensive plant growth facilities (including class 2 containment glass houses) and an extensive imaging suite. The Biomedical Sciences Research Complex at St Andrews is an interdisciplinary research centre where biological, chemical, physical and medical laboratories collaborate under one roof, focusing on host-pathogen interactions. Together, the institutions will provide a highly stimulating and interactive environment with free access to career planning and transferable skills programmes.
Interested candidates are strongly encouraged to contact Dr Tilsner ([email protected]
), Dr Hemsley ([email protected]
), or Dr Roberts ([email protected]
) before applying.