Under supervision of: Kilian Duijts (kilian.duijts@wur.nl)

Aim: Generating and characterizing Crispr knockout mutants of salt-induced root developmental genes.

Soil salinity is an increasing thread to agriculture worldwide. To develop crops that are better adapted to salt stress we need to understand in what way salinity impacts the development of root systems. In this thesis/internship you will be studying (a number of) genes that are involved in the regulation of root branching during salinity stress.

In previous experiments we have identified a salt induced transcription factor (SIT1) that is involved in lateral root development during salt stress. In vitro experiments have revealed a number of potential interaction partners (pip) of SIT1. You will employ use a Crispr Cas9 system to generate knockout(s) of these pip(s), create complementation constructs and phenotype previously generated/related (Crispr) mutants. You will gain experience with various molecular techniques as well as the characterization of Root System Architecture in Arabidopsis.

Are you interested in using molecular techniques to investigate plant developmental regulation and stress tolerance? Get in touch!

Applied techniques:

  • Cloning
  • Crispr knockout generation
  • genotyping
  • qPCR
  • phenotyping of root system architecture
  • microscopy
  • data analysis in R/python

Further reading:

  • Stuttmann, J. et al. Highly efficient multiplex editing: one-shot generation of 8× Nicotiana benthamiana and 12× Arabidopsis mutants. Plant J. 106, 8–22 (2021), doi: 10.1111/tpj.15197
  • Julkowska, M. M. et al. Capturing arabidopsis root architecture dynamics with root-fit reveals diversity in responses to salinity. Plant Physiol. 166, 1387–1402 (2014), doi: 10.1105/tpc.16.00680
  • Karlova, R., Boer, D., Hayes, S. & Testerink, C. Root plasticity under abiotic stress. Plant Physiol. 187, 1057–1070 (2021), https://dx.doi.org/10.1093/plphys/kiab392.
  • Lavenus, Julien, et al. “Lateral root development in Arabidopsis: fifty shades of auxin.” Trends in plant science 18.8 (2013): 450-458 https://dx.doi.org/10.1016/j.tplants.2013.04.006
  • Banda, Jason, et al. “Lateral root formation in Arabidopsis: a well-ordered LRexit.” Trends in plant science 24.9 (2019): 826-839, doi:10.1016/j.tplants.2019.06.015.