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Divide and conquer: Uncovering the mechanism of vacuole re-organisation during cytokinesis

Principal Supervisor: Professor Lorenzo Frigerio

Secondary Supervisor(s): Dr Joseph McKenna

University of Registration: University of Warwick

BBSRC Research Themes: Sustainable Agriculture and Food (Plant and Crop Science)

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Deadline: 4 January, 2024


Project Outline

The diversity of plant morphology is spectacular, from immense heights of the Californian redwood to the herbs found on our dining tables. These differing morphologies are driven by controlled and coordinated plant cell growth. The vacuole is a hallmark of the plant cell, this single organelle provides the outward pushing turgor pressure fundamental for growth of every plant cell on the planet. As plants are the basis of our planetary food supply, understanding how this vacuole is regulated and remodelled will have direct implications for sustainably enhancing agriculture.  

Professor Lorenzo Frigerio and Dr Joe McKenna have studied a specialised type of vacuole in plant embryos, the protein storage vacuole. We have identified how a process of phase separation drives specialised vacuole biogenesis in plant embryos [1]. This work has led us to ask a simple follow-up question, how is the vacuole regulated during cell division?  

Previous work by other groups has demonstrated that during cell division the vacuole reduces in size by 80% and fragments allowing the spindle fibres to form, chromosomes to separate and dividing cells to organise themselves [2]. However, 15-20 minutes after cell division the vacuole is completely remodelled and expanded to its normal size in both daughter cells. The aim of this project is to determine the molecular mechanism driving rapid shrinkage, fragmentation, merging and subsequent re-expansion of the vacuole during cell division.  

The successful candidate will become a highly trained cell biologist proficient in molecular cloning, confocal imaging, super-resolution Structured Illumination Microscopy (SIM) and proximity labelling proteomics. All these techniques are available in house in the School of Life Sciences at the University of Warwick in our world class imaging and proteomics facility and are readily used in the host lab.

References

  1. Kusumaatmaja H, May AI, Feeney M, et al (2021) Wetting of phase-separated droplets on plant vacuole membranes leads to a competition between tonoplast budding and nanotube formation. Proc Natl Acad Sci USA 118:e2024109118. https://doi.org/10.1073/pnas.2024109118
  2. Seguí-Simarro JM, Staehelin LA (2006) Cell cycle-dependent changes in Golgi stacks, vacuoles, clathrin-coated vesicles and multivesicular bodies in meristematic cells of Arabidopsis thaliana: A quantitative and spatial analysis. Planta 223:223–236. https://doi.org/10.1007/s00425-005-0082-2

Techniques

  • MoClo Golden gate multi gene assembly.
  • Confocal live cell imaging.
  • Structured Illumination Microscopy (SIM) with photomanipulation.
  • Proximity labelling proteomics.