Vascular tissues are essential for the development of any higher organism and in plants, dead tracheary element cells (TEs) rely on secondary cell wall thickenings on the cell sides to maintain the water and minerals sap flow to all organs (Pesquet and Lloyd, 2011; Ménard and Pesquet, 2015). The specific organization of TE cell walls into distinct patterns is controlled by underlying microtubule bundles which are organized by specific microtubule associated/interacting proteins (Pesquet et al., 2010; Derbyshire et al., 2015). The aim of the present master thesis is to unravel the function of candidate microtubule associated/interacting proteins identified recently (Derbyshire et al., 2015).

Supervision: Edouard Pesquet – edouard.pesquet@su.se

Experimental approach: This is a genetic, molecular and cell biology project which will include (i) genetic analysis of insertional mutants, (ii) gene constructs design for stable genetic transformation, (iii) phenotypic characterization of mutants and transgenic plants and (iv) biochemical characterization of candidate microtubule associated/interacting protein.

Techniques involved: Training in this project (genetic, molecular and cell biology) will include (i) DNA and RNA extraction and characterization, (ii) PCR and RT-PCR, (iii) bio-informatic sequence analysis (Blast, alignment, phylogeny), (iv) Gateway cloning and associated microbiology techniques, (v) stable genetic transformation, (vi) phenotypic analysis of mutant and transgenic plants (β-glucuronidase promoter analysis,…) and (vii) purification of candidate microtubule associated/interacting protein.
 

Impact of microtubule organization in sap conduit cells of Arabidopsis thaliana plants. Single cell sap conduits exhibiting different cell wall pattern including spiral structure (A) or pitted organization (B). Bar = 10 μm.
Impact of microtubule organization in sap conduit cells of Arabidopsis thaliana plants. Single cell sap conduits exhibiting different cell wall pattern including spiral structure (A) or pitted organization (B). Bar = 10 μm.

 

References:

  • Ménard D. and Pesquet E. (2015) Cellular interactions during tracheary elements formation and function. Current Opinion in Plant Biology, 23, 109-115.
  • Derbyshire,P., Ménard D. Green P., Saalbach G., Lloyd CW. and Pesquet E. (2015) Proteomic analysis of microtubule interacting proteins over the course of xylem tracheary element formation in Arabidopsis. Plant Cell, in press
  • Pesquet E. and Lloyd C (2011) Microtubules, MAPs and xylem formation, In Springer Books: The Plant Cytoskeleton, Edited by Bo Liu, Advances in Plant Biology 2, Chapter 13 (Edited by B. Liu), 277-306.
  • Pesquet E., Korolev A.V., Calder G. and Lloyd C.W. (2010) The microtubule-associated protein AtMAP70-5 regulates secondary wall patterning in Arabidopsis wood cells. Current Biology, 20, 744-749.