PhD Topics

Characterization of components of cytokinin metabolism involved in shoot regeneration.

Supervisor: Mgr. Markéta Pernisová, Ph.D.        

Annotation:

Plants, unlike animals, possess a unique developmental plasticity that enables them to develop new organs throughout their entire life. This ability requires the presence of regulators that trigger specific spatiotemporal changes in developmental programs. In de novo shoot regeneration, cytokinins are the prime candidates for the signal that determines the shoot identity. The process is accompanied by changes in the metabolism, signalling and by differential regulation of gene expression.

The project will aim to elucidate the role and the impact of cytokinins during de novo shoot regeneration. A wide range of molecular biology methods will be used, mainly CRISPR/Cas9 system to generate mutants in cytokinin metabolic enzymes, as well as several cloning methodologies (Gibson assembly, Gateway etc.), in vitro cultivation, luciferase assay or microscopy and image analysis.

Requirements on candidates:

coursework in molecular biology

Recommended literature:

• Pernisová M, Grochová M, Konečný T, Plačková L, Haruštiaková D, Kakimoto T, Heisler MG, Novák O, Hejátko J: Cytokinin signalling regulates organ identity via AHK4 receptor in Arabidopsis. Development 2018, Jul 20;145(14). doi: 10.1242/dev.163907.
• Pernisová M, Vernoux T: Auxin does the SAMba: Auxin signalling in the shoot apical meristem. Cold Spring Harb Perspect Biol 2021 Apr 26; a039925. doi: 10.1101/cshperspect.a039925.
• Capdeville N, Merker L, Schindele P, Puchta H: Sophisticated CRISPR/Cas tools for fine-tuning plant performance. J Plant Physiol. 2021 Feb;257:153332. doi: 10.1016/j.jplph.2020.153332.

Keywords: Arabidopsis, CRISPR/Cas, crops, cytokinins, in vitro regeneration, shoot