Research

The focus of my research is the cell biology of plant-pathogen interactions, in particular between the notorious oomycete pathogen Phytophthora infestans, the causal agent of the devastating potato late blight, and its hosts. Pathogens manipulate plant defence responses on multiple levels, suggesting complex exchanges of signals between host and pathogen. I work closely with Paul Birch (of Dundee University Plant Sciences, based at the James Hutton Institute) and Steve Whisson and other members of the strong Phytophthora team in Dundee and we are part of the Dundee Effector Consortium which encourages collaboration and exchange between teams working on different pathogen/pest-plant interactions. We have fruitful collaborations with Phytophthora researchers in New Zealand, South Africa and China.

P. infestans is a hemi-biotroph and the biotrophic phase of growth is essential for infection. It represents a point of vulnerability and is the stage at which the pathogen must exert the most control over the host defences. To do this the pathogen secretes a broad range of pathogenicity factors, referred to as effectors, many of which are protein. A key group of these are the RXLR (+/-EER) effectors (their conserved motif is similar to the one involved in protein translocation from the malaria parasite into host cells). The formation of haustoria, finger-like projections that breach the plant cell wall to make contact with the cell membrane, is crucial for the biotrophic phase of infection. We have shown that effectors are secreted at haustoria and that RXLR effectors are translocated into host cells.

Characterising the activities of several RXLR effectors revealed how the pathogen targets a broad range of host pathways that contribute to plant defence, often using multiple effectors to target different elements within the same pathway. We are also investigating how effectors are delivered to host cells; what is the secretion pathway, the entry route and how effectors reach their final destinations.

One of the first effectors we examined, PiAVR3a, has 14 candidate host protein interactions. The complexities of the interaction between PiAVR3a and the E3 ligase CMPG1 alone took several years to unravel. The nuclear-targeted effector Pi04314, one of the most highly expressed effectors forms a host-pathogen holoenzyme with protein phosphatase type 1 catalytic domain (PP1c), exploiting the phosphatase activity to benefit infection. Similarly the nucleolar-associated effector Pi04089 exploits the activity of a KH type of RNA binding protein (StKRBP1), Pi02860 targets a predicted potato E3 ligase NPH3/RPT2-like 1 (StNRL1) and Pi17316 uses MAP3K StVIK1. Thus StPP1c’s the StKRBP1, StNRL1 and StVIK1 are all susceptibility (S) factors. This reveals the potential of S factors as novel targets for development of plant resistance.

Initially we showed that RXLR-EER motif is required for translocation of PiAVR3a into the plant cytosol using GUS fusions. Subsequently, using effectors that we found were nuclear localised when expressed in plant cells, we managed to observe effector translocation in live cells with fluorescent protein fusions. Inhibitor studies revealed that apolastic effectors and other pathogenicity proteins were conventionally secreted while the RXLR efectors tested were secreted by a different pathway, i.e. non-conventionally secreted. We are investigating how RXLR effectors are targeted to this non-conventional route and how that results in them being translocated into host cells. We have shown that uptake of a range of RXLR effectors by plant cells involves clathrin-mediated endocytosis. The next questions we have are whether specific signals are involved in this endocytosis and how are effectors released from endosomes to reach their host targets in the cytosol or other locations.

The imaging and cell biology facilites and capabilites at The James Hutton Institute are managed by Alison Roberts, Michael Porter and Sean Chapman in addition to their own research activities. High quality imaging work is not possible without great training, ongoing support and facility management.

Publications

Journals

Books / chapters

  • Boevink, P.; McLellan, H.; Bukharova, T.; Engelhardt, S.; Birch P.R.J. (2014) In vivo protein-protein interaction studies with BiFC: conditions, cautions, and caveats., In: Birch, P.R.J., Jones, J.T. & Bos, J.I.B. (eds.). Plant-Pathogen Interactions: Methods and Protocols. Methods in Molecular Biology, Humana Press, New York, 2nd ed. Volume 1127, Chapter 6, 81-90.
  • Boevink, P.; Birch, P.R.J.; Whisson, S.C. (2011) Imaging fluorescently tagged Phytophthora effector proteins inside infected plant tissue., In: McDowell, J.M. (ed.). Methods in Molecular Biology Plant Immunity: Methods and Protocols. Humana Press, Totowa, New Jersey, USA, Chapter 712, pp195-209.
  • Oparka, K.J.; Boevink, P. (2004) Techniques for imaging intercellular transport., In: Plasmodesmata (ed. K.J. Oparka). Blackwell Publishing, Oxford, pp241-262
  • Hawes, C.; Boevink, P.; Moore, I. (2000) GFP in plants., In: Fluorescence Microscopy of Proteins: A Practical Approach (ed. V. Allen). Irl Press, pp163-177
  • Neuhaus, J.; Boevink, P. (2000) The green fluorescent protein (GFP) as a reporter in plant cells., In: Plant Cell Biology: A Practical Approach (eds. C.L. Hawes and B. Satiat-jeunemaitre). Oxford University Press, Oxford, pp127-142.

Technical / contract reports

  • Whisson, S.C.; Boevink, P.; Pritchard, L.; Avrova, A.O.; Morales, J.; Birch, P.R.J. (2006) Effector protein translocation in the Phytophthora infestans – potato infection., Annual Report of the Scottish Crop Research Institute for 2006, pp24-25.
  • Latijnhouwers, M.; Boevink, P.; Hawes, C.L.; Oparka, K.J. (2004) In search of plant Golgi matrix proteins., Annual Report of the Scottish Crop Research Institute for 2003/2004, 131-132.
  • Boevink, P.; Calcutt, J.; Roberts, A.G.; Oparka, K.J. (2004) Real time studies of cell-to-cell communication., Annual Report of the Scottish Crop Research Institute for 2003/2004, 129-130.
  • Gillespie, T.; Toth, R.; Haupt, S.; Boevink, P.; Roberts, A.G.; Chapman, S.N.; Oparka, K.J. (2001) A DNA-shuffled movement protein enhances virus transport by evasion of a host-mediated degradation pathway., Annual Report of the Scottish Crop Research Institute for 2000/2001, pp107-111.
  • Roberts, A.G.; Santa, Cruz, S.; Boevink, P.; Roberts, I.M.; Sauer, N.; Oparka, K.J. (1999) The sink-source transition in leaves – new insights., Annual Report of the Scottish Crop Research Institute for 1998/99, 76-79.
  • Santa, Cruz, S.; Boevink, P.; Duncan, G.; Roberts, A.G.; Prior, D.A.M.; Oparka, K.J. (1998) Probing the virus long-distance tranport pathway., Annual Report of the Scottish Crop Research Institute for 1997/98, 114-117.
  • Boevink, P.; Hawes, C.L.; Prior, DA.M.; Santa, Cruz, S.; Oparka, K.J. (1998) New insights into the plant secretory pathway using virus-delivered green fluorescent protein., Annual Report of the Scottish Crop Research Institute for 1997/98, 67-70.

Conference papers

  • Wang, H.; Wang, S.; Wang, W.; Welsh, L.; Gierlinski, M.; Whisson, S.; Hemsley, P.; Boevink, P.; Birch, P. (2022) Entering via the front door Cytoplasmic effectors are taken into host cells via endocytosis, 52nd Congress of the Southern African Society for Plant Pathologists (SASSP), 1-3 August 2022, Pretoria.
  • Boevink, P.C.; Wang, S.; Pham, J.; Turnbull, D.; Welsh, L.; Whisson, S.C.; Birch, P.R.J. (2019) The oomycete RXLR motif determines non-conventional secretion for effector delivery., IS-MPMI Molecular Plant Microbe Interactions, XVIII Congress, Glasgow, 14-18 July 2019.
  • He ,Q.; Naqvi, S.; McLellan, H.; Boevink, P.C.; Champouret, N.; Hein, I.; Birch, P.R.J. (2018) A plant pathogen effector utilizes host Susceptibility factor NRL1 to degrade the immune regulator SWAP70, Oomycete Molecular Genetics Network Meeting, Shandong Agricultural University, Taian, China, 8-12 April 2018.
  • Wang, S.; Boevink, P.C.; Whisson, S.C.; Birch, P.R.J. (2018) How are effectors secreted and delivered to the host?, Oomycete Molecular Genetics Network Meeting, Shandong Agricultural University, Taian, China, 8-12 April 2018.
  • Boevink, P.; Wang, X.; McLellan, H.; Armstrong, M.; Bukharova, T.; Birch, P.R.J. (2014) Targeting of PP1c isoforms by an RXLR effector., Oomycete Molecular Genetics Network Meeting, Norwich, July 2014.
  • Stevens, L.; Chapman, S.; Boevink, P.; Engelhardt, S.; Chen, X.; Birch, P.; Hein, I. (2014) Shuffling resistance genes to protect solanaceous plants., British Society for Plant Pathology Presidential Meeting, University of St Andrews, 1-2 September 2014.
  • Breen, S.; Gilroy, E.M.; Armstrong, M.R.; Morales, J.G.; Hein, I.; Douglas, E.; Boevink, P.; McLellan, H.; Randall, E.; Tian, Z.D.; Avrova, A.O.; Pritchard, L.; Lokossou, A.A.; Govers, F.; van der Vossen, E.A.G.; Vleeshouwers, V.G.A.A.; Brunner, F.; Whisson, S.C.; Birch, P.R.J. (2010) Avr2, an RXLR effector from Phytophthora infestans., Oomycete Molecular Genetics Network Congress, Toulouse, France, 6-8 June 2010.
  • Armstrong, M.R.; Boevink, P.; Bukharova, T.; McLellan, H.; Ramos, M.; Gilroy, E.M.; Pritchard, L.; Whisson, S.C.; Beynon, J.; Birch, P.R.J. (2010) Identifying candidate host ‘targets’ of Phytophthora infestans translocated RXLR proteins., Oomycete Molecular Genetics Network Congress, Toulouse, France, 6-8 June 2010.
  • Engelhardt, S.; Armstrong, M.R.; Boevink, P.; Ramos, M.; Gilroy, E.M.; Birch, P.R.J. (2010) R3a-mediated recognition of AVR3a: Cell biological and biochemical characterisation., Oomycete Molecular Genetics Network Congress, Toulouse, France, 6-8 June 2010.
  • Gilroy, E.M.; Breen, S.; Hein, I.; Tian, Z.D.; Armstrong, M.R.; Douglas, E.; Lokossou, A.A.; Boevink, P.; McLellan, H.; Randall, E.; Morales, J.G.; Avrova, A.O.; Pritchard, L.; van der Vossen, E.A.G.; Whisson, S.C.; Birch, P.R.J. (2009) Amino acid polymorphisms upstream of the RXLR motif in Phytophthora infestans effector Avr2 condition R gene recognition., The Fungal Cell, British Mycological Society Meeting, Westpark Conference Centre, Dundee, 1-4 September 2009.
  • Gilroy, E.M.; Armstrong, M.R.; Bos, J.; Taylor, R.; Morales, J.; Hein, I.; Douglas, E.; Boevink, P.; Avrova, A.O.; Breen, S.; van West, P.; Pritchard, L.; Whisson, S.C.; Sadanandom, A.; Kamoun, S.; Birch, P.R.J. (2009) Functional analysis of Phytophthora infestans RXLR effectors Avr2 and Avr3a., Fungal Genetics Conference, Asilomar Conference Grounds, Pacific Grove, California, USA, 17-22 March 2009.
  • Birch, P.R.J.; Armstrong, M.R.; Gilroy, E.M.; Bos, J.; Taylor, R.M.; Morales, J.G.; Hein, I.; Douglas, E.; Boevink, P.; Breen, S.; Pritchard, L.; Whisson, S.C.; Sadanandom, A.; Kamoun, S. (2009) Localisation and function of Phytophthora infestans RXLR effectors and their host targets., XIV MPMI Conference, Montreal, Canada, 19-23 July 2009.
  • Boevink, P.; Ramos, M.; Armstrong, M.R.; Allen, R.; Gilroy, E.M.; Donovan, S.; Rehmany, A.; Beynon, J.; Birch, P.R.J. (2009) Evolutionarily distinct RXLR effectors from distantly related oomycetes target the plant exocyst., 14th International Congress on Molecular Plant-Microbe Interactions, Quebec, Canada, 19-23 July 2009.
  • Grouffaud, S.; Boevink, P.; Avrova, A.O.; van West, P.; Birch, P.R.J.; Whisson, S.C. (2008) Translocation of effector proteins from the oomycete Phytophthora infestans into plant cells., 9th European Conference on Fungal Genetics, Edinburgh, 5-8 April 2008.
  • McLellan, H.; Gilroy, E.M.; Hein, I.; Boevink, P.; Birch, P.R.J.; Loake, G. (2007) The roles of cathespin B-like proteases in plant disease resistance., XIII International Congress on Molecular Plant-Microbe Interactions, Sorrento, Italy, 22-26 July 2007.
  • Avrova, A.O.; Boevink, P.; Grenville-Briggs, L.J.; van West, P.; Birch, P.R.J.; Whisson, S.C. (2007) Identification of novel pathogenicity factors in the late blight pathogen, Phytophthora infestans., XIII International Congress on Molecular Plant-Microbe Interactions, Sorrento, Italy, 22-26 July 2007.
  • Whisson, S.C.; Boevink, P.; Moleleki, L.N.; Avrova, A.O.; Morales, J.G.; Gilroy, E.M.; Armstrong, M.R.; Toth, I.K.; Pritchard, L.; Birch, P.R.J. (2007) A translocation signal for delivery of oomycete effector proteins into host plant cells., XIII International Congress on Molecular Plant-Microbe Interactions, Sorrento, Italy, 22-26 July 2007.
  • Gilroy, E.M.; Venter, E.; McLellan, H.; Hein, I.; Hrubikova, K.; Holeva, M.C.; Boevink, P.; Loake, G.J.; Lacomme, C.; Birch, P.R.J. (2006) Inducible disease resistance involves an apoplastic cysteine protease Cathepsin B., Proceedings of IOBC/wprs Joint Working Group Meeting: Breeding for Inducible Resistance against Pests and Diseases, Heraklion, Crete, 27-29 April 2006.
  • Carvalho, C.; Latijnhouwers, M.; Osterrieder, A.; Boevink, P.; Oparka, K.J.; (2005) Screening for plant Golgi-associated proteins., Mini-Symposium on Improving the Plant Secretory System for Nutrition and Health, Grasmere, 23-26 May 2005.
  • Latijnhouwers, M.; Hawes, C.L.; Oparka, K.J.; Carvalho, C.; Boevink, P. (2005) Plant golgi matrix proteins., Mini-Symposium on Improving the Plant Secretory System for Nutrition and Health, Grasmere, 23-26 May 2005.
  • Haupt, S.; Wright, K.M.; Cowan, G.H.; Boevink, P.; Gillespie, T.; Ziegler, A.; Roberts, A.G.; Chapman, S.N.; Oparka, K.J.; Torrance, L. (2005) Plant virus proteins traffic in the endomembrane system., Mini-Symposium on Improving the Plant Secretory System for Nutrition and Health, Grasmere, 23-26 May 2005.
  • Boevink, P.; Wright, K.M.; Chapman, S.N.; Oparka, K.J. (2005) Tools for protein tracking., Plant Sciences Seminar and Workshop; Applications for Fluorescence Microscopy, Rothamsted Research, Harpenden, 11-15 April 2005.
  • Boevink, P.; Chapman, S.N.; Bell, K.; Jackson, K.; Wright, K.M.; Latijnhouwers, M.; Gillespie, T.; Oparka, K.J.; Roberts, A.G. (2005) Investigating protein location, motion and function in living plant cells., Biotechnology, Havana, Cuba, 27 November – 2 December 2005.
  • Boevink, P.; Wright, K.M.; Roberts, A.G.; Wood, N.; MacKenzie, K.; Oparka, K.J. (2005) Towards a new model for TMV movement., Plasmodesmata 2004, Monterey, California, USA, 17-21 August 2004.
  • Roberts, A.G.; Boevink, P.; Calcutt, J.; Chapman, S.N.; Wright, K.M.; Oparka, K.J. (2004) New fluorescent proteins light up plant intercellular communication., SEB Plant Frontiers – The Visible Cell Biosensors and Bioreceptors Session, Exeter 3-6 July 2004.
  • Wright, K.M.; Wood, N.T.; Roberts, A.G.; Boevink, P.; Gillespie, T.; MacKenzie, K.; Oparka, K.J. (2004) Plasmodesmal targeting of TMV movement protein utilises the ER/actin network., Plasmodesmata 2004, Monterey, California, USA, August 17-21 2004.
  • Boevink, P.; Oparka, K.J. (2004) Real-time studies of plasmodesmatal gating in developing leaf cells., Plasmodesmata 2004, Monterey, California, USA, 17-21 August 2004.

Conference posters / abstracts

  • Chapman, S.; Stevens, L.; Boevink, P.; Engelhardt, S.; Alexander, C.; Champouret, N.; Birch, P.; Hein, I. (2014) Artificial evolution of the potato resistance gene R3a to detect the virulent form of AVR3a., British Society for Plant Pathology Presidential Meeting, University of St Andrews, 1-2 September 2014. (Poster)
  • Birch, P.R.J.; Boevink, P.C.; Gilroy, E.M.; Hein, I.; Pritchard, L.; Whisson, S.C. (2012) Effectors from the potato late blight pathogen: their roles in suppressing or triggering the plant immune system., EPS Autumn-school, Wageningen, 6 March 2012.
  • Birch, P.R.J.; Boevink, P.C.; Gilroy, E.M.; Hein, I.; Pritchard, L.; Randall, E.; Whisson, S.C. (2011) Effectors from the potato late blight pathogen: their roles in suppressing or triggering the plant immune system., British Mycological Society Meeting, Exeter, September 2011.
  • Boevink, P.; Bukharova, T.; McLellan, H.; Armstrong, M.R.; Gilroy, E.M.; Birch, P.R.J. (2010) Imaging plant-pathogen interactions: Phytophthora infestans RXLR effector localisation in planta., 9th International Mycological Congress,The Biology of Fungi, Edinburgh, 1-6 August 2010 (Poster).
  • Gilroy, E.M.; Breen, S.; Morales, J.G.; Lokossou, A.A.; Ramos, B.; Armstrong, M.R.; Hein, I.; Boevink, P.; Pritchard, L.; van der Vossen, E.A.G.; Vleeshouwers, V.G.A.A.; Whisson, S.C.; Birch, P.R.J. (2009) Cloning and functional analysis of Phytophthora infestans RXLR effector, Avr2., 22nd New Phytologist Symposium on Effectors in Plant-Microbe Interactions, INRA Versailles Research Centre, Paris, France, 13-16 September 2009 (Poster).
  • Avrova, A.O.; Boevink, P.; Grenville-Briggs, L.J.; van West, P.; Birch, P.R.J.; Whisson, S.C. (2007) Transient gene silencing – a step forward in identifying novel pathogenicity factors in the late blight pathogen., BSPP Presential Meeting, University of Bath, 12-14 September 2007 (Poster).
  • Roberts, A.G.; Wright, K.M.; Wood, N.T.; Gillespie, T.; Boevink, P.; Haupt, S.; Oparka, K.J. (2003) Intercellular targeting of viral movement protein to plasmodesmata., Abstracts of the International Conference on Phloem Transport, University of Bayreuth, Germany, 31 August – 5 September 2003.
  • Wright, K.M.; Wood, N.T.; Gillespie, T.; Boevink, P.; Roberts, A.G.; Oparka, K.J. (2003) Intercellular viral movement protein targeting to plasmodesmata., Abstracts of the Seventh International Botanical Microscopy Meeting, Lisbon, Portugal, 12-17 April 2003.
  • Roberts, I.M.; Boevink, P.; Roberts, A.G.; Sauer, N.; Reichel, C.; Oparka, K.J. (2001) Dynamic changes in the frequency and architecture of plasmodesmata during the sink-source transition in tobacco leaves., Abstracts of the 29th Scottish Microscopy Symposium, Glasgow, 14 November 2001, 12.
  • Roberts, I.M.; Boevink, P.; Roberts, A.G.; Sauer, N.; Reichel, C.; Oparka, K.J. (2001) Dynamic changes in the frequency and architecture of plasmodesmata during the sink-source transition in tobacco leaves., Abstract for the ‘Plasmodesma 2001’ Meeting, Cape Town, South Africa, 19-24 August 2001, 6-8.
  • Boevink, P.; Haupt, S.; Toth, R.L.; Roberts, A.G.; Oparka, K.J. (2001) Cell-cell movement of TMV in the absence of microtubules., Abstract for the ‘Plasmodesma 2001’ Meeting, Cape Town, South Africa, 19-24 August 2001, 57.
  • Gillespie, T.; Toth, R.L.; Haupt, S.; Boevink, P.; Roberts, A.G.; Chapman, S.N.; Oparka, K.J. (2001) A DNA-shuffled movement protein enhances transport of TMV by evasion of a host degradation pathway., Abstract for the ‘Plasmodesma 2001’ Meeting, Cape Town, South Africa, 19-24 August 2001, 38-40.
  • Roberts, A.G.; Boevink, P.; Santa Cruz, S.; Sauer, N.; Oparka, K.J. (2000) Functional dynamics in sink-source transition leaves., Abstracts of the 28th Scottish Microscopy Symposium, Moredun Institute, Edinburgh, 15 November 2000, 6.
  • Roberts, A.G.; Boevink, P.; Santa Cruz, S.; Sauer, N.; Oparka, K.J. (2000) Using GFP to study the plasmodesmatal size exclusion limit in sink and source tissues., Abstracts of the Annual SEB Meeting, Exeter, 27-31 March 2000.
  • Roberts, A.G.; Boevink, P.; Santa Cruz, S.; Sauer, N.; Oparka, K.J. (2000) A study of plasmodesmata during the sink-source transition: GFP labelling reveals dynamic changes in architecture and function., Abstracts of the ICHC, York University, York, 3-8 September 2000, 45.
  • Roberts, A.G.; Boevink, P.; Santa Cruz, S.; Sauer, N.; Oparka, K.J. (1999) Plasmodesmata and the sink-source transition – new insights., Abstracts of the International Congress on Assimilate Transport and Partitioning, Newcastle, Australia, August 1999, 121.
  • Brandizzi, F.; Boevink, P.; Oparka, K.J.; Hawes, C.L. (1999) Endoplasmic reticulum is highlighted in higher plants., Abstracts of the BSCB Meeting, Bristol, 20-22 September 1999.
  • Boevink, P.; Roberts, A.G.; Brandizzi, F.; Saint-Jore, C.; Oparka, K.J.; Hawes, C.L. (1999) Golgi bodies track along the endoplasmic reticulum in plants., Abstracts of the BSCB Meeting, Bristol, 20-22 September 1999.
  • Hawes, C.L.; Saint-Jore, C.; Moore, I.; Oparka, K.J.; Santa Cruz, S.; Boevink, P. (1999) Green fluorescent protein to study the dynamics of the Golgi and the plant secretory pathway., Abstracts of the Italian Botanical Society, Ferrera, Italy, 1999.
  • Boevink, P.; Lacomme, C.; Wright, K.M.; Hawes, C.L.; Santa Cruz, S.; Oparka, K.J. (1999) GFP and plant-virus vectors as tools for elucidating protein localisation and function., Abstracts of Molecular Biology and Fungal Pathogens, Gregynog, Wales, July 1999.