Dr P Hemsley, Dr J Tilsner, Dr A Roberts
No more applications being accepted
Competition Funded PhD Project (European/UK Students Only)
About the Project
Background
Plant viruses are major crop pathogens with a global agro-economic impact. The Tilsner lab has recently made major advances in understanding how viruses transport their genome from infected to naïve cells through plasmodesmata, cell-cell-connecting membrane-lined channels. Using a novel method for detecting protein-protein interactions in vivo developed by the Hemsley lab, this PhD project will characterize a putative membrane-anchored contractile protein machinery at plasmodesmata that is required for virus transport and more broadly for the regulation of intercellular communication in plants.
Project details
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 in vivo detection of protein-protein interactions to identify new components of a putative actin-related contractile system at plasmodesmata. The plasmodesmatal localization of new candidates will be confirmed by live cell fluorescence microscopy (confocal and super-resolution). 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. Parallel experiments will be also performed in Nicotiana benthamiana using transient expression studies and gene silencing.
The PhD project will provide in-depth training in several state-of-the-art technologies such as: molecular biology including restriction-free cloning techniques (Gibson assembly and Gateway); protein biochemistry and mass spectrometry; advanced light (confocal, FRAP, SIM super-resolution) and electron microscopy (tomography); transient and stable plant transformation; handling of genetically modified crop pathogens in class 2 containment laboratories and glass houses. After initial training in required methods, the student will be encouraged to take intellectual leadership of the project from an early stage. They will have ample opportunity to present their work at internal seminars as well as at international conferences in the second half of the PhD, and be encouraged to write portions or first drafts of resulting papers.
The James Hutton Institute is a world-leading plant research center with particular focus on molecular plant-pathogen interactions. It has comprehensive plant growth facilities (including class 2 containment glass houses) and extensive plant imaging facilities. 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. It provides state-of-the-art laboratories and equipment, and a highly international work environment. Together, the two institutions will provide a highly stimulating and interactive environment for the PhD student. Both also offer free access to career planning and transferable skills programmes. The University of St Andrews is the oldest university in Scotland and consistently ranked as amongst the United Kingdom’s top ten universities. Currently, it is UK University of the Year for Student Experience in The Times and Sunday Times Good University Guide 2018.
Funding Notes
The studentship is funded under the James Hutton Institute/University Joint PhD programme, in this case with the University of St Andrews, for 3.5 years. Applicants should have a first-class honours degree in a relevant subject or a 2.1 honours degree plus Masters (or equivalent).Shortlisted candidates will be interviewed in Jan/Feb 2019. A more detailed plan of the studentship is available to candidates upon application. Funding is available for European applications, but Worldwide applicants who possess suitable self-funding are also invited to apply. Interested candidates are strongly encouraged to contact Dr Tilsner ([Email Address Removed]) or Dr Hemsley ([Email Address Removed]) before applying.
References
Tilsner J, Nicolas W, Rosado A, Bayer EM (2016) Staying tight: Plasmodesmal membrane contact sites and the control of cell-to-cell connectivity in plants. Annu. Rev. Plant Biol. 67, 337-364. doi: 10.1146/annurev-arplant-043015-11184
Tilsner J, Linnik O, Louveaux M, Roberts IM, Chapman SNC, Oparka KJ (2013). Replication and trafficking of a plant virus are coupled at the entrances of plasmodesmata. J. Cell Biol. 201: 981-995. doi:10.1083/jcb.201304003