Project properties
Title Functional characterization of flavor-related genes in pea (Pisum sativum L.) using CRISPR/Cas9 and genetic transformation
Group Plant Breeding, Laboratory of
Project type thesis
Credits 30-39
Supervisor(s) Prof.dr. Luisa Trindade and Ornela Bocova
Examiner(s) Prof.dr. Luisa Trindade
Contact info ornela.bocova@wur.nl ; luisa.trindade@wur.nl
Begin date 2024/02/01
End date 2025/06/30
Description In the protein transition era, pulses and in particular pea, are regaining interest for their high protein level and adaptation to the European climate. However, food application of pulse ingredients as meat and dairy replacers is hindered by their off-flavor. Off-flavors result from a combination of volatile and non-volatile compounds responsible for the aroma/odor and taste. In this project we aim to understand the genetic determinants of flavor compounds in pea and to develop tools for breeding varieties without or less off-flavors.
The student will work on the functional characterization of key genes involved in pathways leading to off-flavor compounds. We will target and Knock Out genes of interest using targeted mutagenesis methods like Crispr/Cas and Agrobacterium-mediated plant transformation.
The student will screen protocols, and perform transformation experiments with constructs containing selection and reporter genes. The work will also involve the generation of constructs to Knock Out (CRISPR/Cas) candidate genes.
Used skills Plant Cell and Tissue culture
Plant Transformation using Agrobacterium tumefaciens
Validation of transformation by phenotype (microscopy) and PCR
Generation of constructs for candidate genes using CRISPR/Cas with Golden Gate Cloning system
Requirements Students that followed several of the following courses are preferred: Plant Cell and Tissue culture, Gene technology, Plant Biotechnology.
Additional courses: Breeding for Stress Tolerance and Quality or Advanced Bioresources.