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Functional analysis of fungal effector proteins in the barley/wheat powdery mildew plant-pathogen interactions.

  • Full or part time
  • Application Deadline
    Applications accepted all year round
  • Self-Funded PhD Students Only
    Self-Funded PhD Students Only

Project Description

Background
Pathogens secrete small proteins called effectors or virulence factors which are important for the successful invasion of the host. These effectors act by compromising the host immunity. Cereal powdery mildews (Blumeria graminis) are economically important pathogens in the UK, affecting main cereal crops such as barley and wheat. Powdery mildews can only grow on “healthy and alive” cells of their targeted host. Thus they have a life style typical of obligate pathogens, which requires the ability of compromising the host resistance mechanisms for successful infection. The sequenced genome of the barley powdery mildew (Spanu et al, Science, 2010) and mass spectrometry based proteomics has allowed to identifying novel Blumeria effector candidates (BECs, Bindschedler et al, MCP, 2009; Bindschedler et al, Methods, 2011). These BEC proteins were expressed mostly in specialised cells of biotrophic fungi. Thus they were suspected to be involved in the infection process. Seven BECs were then experimentally validated as Blumeria effector proteins (Pliego et al. MPMI 2013). Five of these effectors are novel species specific small secreted proteins, some of which only share similarities with wheat powdery mildew proteins. One of the identified Blumeria effector shares similarity to metalloproteases from other fungi.

Aim of the research
Molecular and proteomic approaches will be used to investigate the role of powdery mildew effector proteins of this plant pathogen during infection of barley by its powdery mildew Blumeria graminis f.sp. hordei.

The PhD project will address some of the following objectives:
- When and where are the effectors expressed?
- Is the universal metalloprotease-like effector a virulence factor of other cereal pathogens?
- What are the effectors mode of action?
- Can a novel RNAi (gene silencing) technology be used to target specific effectors and reduce their expression for functional analysis studies and for controlling fungal diseases?
- Which plant proteins are targetted by these effectors? This will be investigated by protein-protein interactions affinity pull-downs. Orthogonal methods will be used to validate putative candidates. Interactor proteins will be identified by mass spectrometry. Barley proteins which are interacting with the Blumeria effectors will be cloned for further investigation (e.g. gene silencing, reciprocal protein interaction, cell localization) towards understanding their role in the infection/resistance process.
- A protein interaction study will also be undertaken for the metalloprotease like protein.In addition, protease preferential cleavage sites will be investigated, including setting up a mass spectrometry based “degradomic” strategy using the whole barley proteome, i.e. by identifying the barley protease target(s). The mode of action of this protein will be further validated using the RNAi gene silencing technique developed in my lab.

Candidate profile
We are looking for a highly motivated student, with interest in molecular plant pathology and qualifications in biology and/or biochemistry. Preference will be given to students with some knowledge and expertise in molecular biology, and/or protein analysis, including proteomics mass spectrometry.

Funding Notes


References

1. C. Pliego et al. 2013. Host-Induced Gene Silencing in barley powdery mildew reveals a class of ribonuclease-like effectors. MPMI. 26: 633-642.
2. LV. Bindschedler et al. 2011. Proteogenomics and in silico structural and functional annotation of the barley powdery mildew Blumeria graminis f. sp. hordei. Methods 54:432-41.
3. PD Spanu, et al. 2010. Genome expansion 1 and gene loss in powdery mildew fungi reveal functional tradeoffs in extreme parasitism. Science 330:1343-1346.
4. LV. Bindschedler et al. 2009. In Planta Proteomics and Proteogenomics of the biotrophic barley fungal pathogen Blumeria graminis f.sp. hordei. Mol Cell Proteomics 8: 2368-81

How good is research at Royal Holloway, University of London in Biological Sciences?

FTE Category A staff submitted: 24.00

Research output data provided by the Research Excellence Framework (REF)

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