Project details – Institut for Plante- og Miljøvidenskab - Københavns Universitet

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Single-cell proteomics to identify plant “gatekeeper”

Main area:
Target group:Biology-Biotechnology, Agriculture
Educational level:Master, Project in practice, Project outside course scope
Project description:

The goal of this project is to identify and characterize the ‘gatekeeper’ that is key for driving the glucosinolate defense compounds into their storage site, the S-cells. Understanding this process will advance basic knowledge and plant biotechnology towards sustainable agriculture.  

Background: In the model plant Arabidopsis thaliana, glucosinolates are stored to high concentrations (>130 mM) in the vacuole of the 1 mm long laticifer-like sulfur-rich cells called the S-cells. An energy-requiring importer is responsible loading the glucosinolates into the vacuole. Single-cell proteomics on S-cells has enabled identification of candidate vacuolar importers. The project is aimed at screening and characterization the candidates in planta and in vitro.  

Activities:

a)     Clone candidate transporter into yeast and plant expression vector

b)     Biochemical characterization of transporter in yeast (e.g. substrate specificity, Km value) and in planta (localization, expression level)

c)     In planta validation with mutants, metabolite profiling and bioassay

Methodologies: molecular biology, heterologous expression in yeast and plant hosts, biochemical characterization of transporters, LC-MS analytical chemistry, transport engineering, plant genotyping, bioinformatics on candidate gene family.

Keywords: transport biology, yeast, plant, metabolite profiling   

Work environments: We expect you to be enthusiastic, dedicated and eager to learn and advance your skills in many disciplines (e.g. research, oral and written presentations). In return, we provide a stimulating, international research environment for you to advance your skills.  

Main supervisor Prof. Barbara Ann Halkier, DynaMo Center of Excellence, PLEN, Frederiksberg. Email: bah@plen.ku.dk

Co-supervisor: Dr.Deyang Xu, Email: dyxu@plen.ku.dk.  

 

Methods used:Single cell sampling; Molecular biology;Metabolite profiling;Proteomics
Keywords:transport biology, defense compounds, cell biology
Supervisor(s): Barbara Ann Halkier;Deyang Xu
Email:bah@plen.ku.dk