| Title of Project |
| Is the function of a key effector protein in the fungal plant pathogen Z. tritici specific to pathogenic lifestyles? |
| This project going to be… |
| Experimental (lab/field) |
| Full Name of Supervisor |
| Luca Steel |
| Institution Department and Address |
| Biointeractions and Crop Protection, Rothamsted Research Harpenden, Herts AL5 2JQ United Kingdom Map It |
| Telephone |
| 07703690350 |
| luca.steel@rothamsted.ac.uk |
| Position held |
| PhD student |
| Full name of the day to day supervisor and/or arrangements for supervision |
| Luca Steel, with additional support from Jason Rudd and Carlos Bayon. |
| Date of Project Commencement |
| 04/07/2022 |
| Duration (weeks) |
| 10 |
| Brief Description of Project |
| Background: Zymoseptoria tritici is a major fungal pathogen of wheat, causing a disease called Septoria Tritici Blotch. This leads to reduced yields and financial losses. Z. tritici secretes an important effector protein, Zt3LysM, during infection of wheat. This allows Z. tritici to mask its own chitin (a surface cell wall component) to evade detection, and to avoid its hyphae being degraded by enzymes secreted by wheat leaves. We have a fungal strain which has been engineered to lack this gene (deleted – “ΔZt3LysM”) which has strongly reduced virulence on wheat leaves. We have also found that many other fungal species with wide-ranging lifestyles contain proteins with similar sequences. But are they able to perform the same function as Zt3LysM? This project will: complete the first stages of an investigation into whether proteins with similar sequences in other fungal species are functional homologues of Zt3LysM. The student will analyse fungal genomes to design and generate constructs for use in a functional complementation assay. To do this, fungal species (with various lifestyles including non-pathogens) of interest containing proteins with similar sequences to Zt3LysM will be identified. Six of these will be chosen for further analysis, to cover at least: one known orthologous protein from a plant pathogen to act as a positive control; two additional plant pathogens (one wheat pathogen and one non-wheat pathogen); one saprophytic species; and one endophytic species. This will ensure a range of fungal lifestyles are investigated. Genes encoding the selected proteins will be cloned and prepared for transformation into the Z. tritici ΔZt3LysM strain. In a related PhD project, these strains will then be tested to see if they can infect wheat plants thereby testing if the genes from these other fungi are “real” LysM effectors. This will involve a literature review, as well as training in PCR and primer design, gel electrophoresis, Gibson assembly and cloning. This will enhance our understanding of a key effector protein and whether its function is specific to pathogenic lifestyles and potentially reveal cryptic (currently unknown) interactions of other fungi with plants. This will also establish a Z. tritici based method for examining the function of proteins in fungal species which are less amenable to laboratory investigation. Other aims: Through this project, the student will learn and use key laboratory skills which are used widely across disciplines, such as cloning, PCRs, gel electrophoresis, micropipetting, microbial culture, and sterile technique. The student will also gain an understanding of plant pathology and molecular biology. Finally, the student will experience life as a scientist – honing skills in recording, analysing and reporting results, attending internal meetings and seminars, and contributing work to a paper for publication. |
| Attach the recommended reading for the project |
| Dean, R., Van Kan, J. A., Pretorius, Z. A., Hammond-Kosack, K. E., Di Pietro, A., Spanu, P. D., Rudd, J. J., Dickman, M., Kahmann, R., Ellis, J., & Foster, G. D. (2012). The Top 10 fungal pathogens in molecular plant pathology. Molecular plant pathology, 13(4), 422. https://doi.org/10.1111/j.1364-3703.2011.00783.x Orton, E. S., Deller, S., & Brown, J. K. (2011). Mycosphaerella graminicola: from genomics to disease control. Molecular plant pathology, 12(5), 413–424. https://doi.org/10.1111/j.1364-3703.2010.00688.x Tian, H, MacKenzie, CI, Rodriguez-Moreno, L, et al (2021). Three LysM effectors of Zymoseptoria tritici collectively disarm chitin-triggered plant immunity. Molecular Plant Pathology, 22(6), 683– 693. https://doi.org/10.1111/mpp.13055 |
