Project properties
Title Proximity labeling in Arabidopsis to identify protein-protein interactions related to aphid and potyvirus resistance
Group Entomology, Laboratory of
Project type thesis
Credits 24-36
Supervisor(s) Janna Damen
Examiner(s) Marcel Dicke
Contact info janna.damen@wur.nl
Begin date 2024/03/01
End date 2025/03/01
Description Background:
Aphids are major crop pests causing direct feeding damage and additionally vector plant pathogens including potyviruses. The current set of aphid management measures is insufficient, raising the need for new sustainable pest management strategies. The two homologous phloem-based resistance proteins SIEVE ELEMENT-LINING CHAPERONE-1 (SLI1) and RESTRICTED TEV MOVEMENT2 (RTM2) confer resistance to aphids and potyviruses respectively, therefore the goal of this research is to identify proteins that interact with SLI1 or RTM2.

The Project:
During your project, you will contribute to unraveling the molecular basis of phloem-based resistance to aphids and potyviruses. We will obtain Arabidopsis lines that express SLI1 or RTM2 fused to a biotin ligase enzyme (TbID). The TbID in these mutant lines will label interacting proteins, which can then be identified by mass spectrometry. During your project, you will focus on the optimization of this proximity labeling system in Arabidopsis. You will use molecular tools to investigate the functionality of the TbID ligase and determine the optimal conditions for biotin feeding to the plants. If time allows, test samples will be processed for mass spectrometry to identify the first interacting peptides.

!!! Since this project is a collaboration between the Laboratories of Entomology and Virology, the majority of your work will be performed at the Laboratory of Virology. However, this might vary depending on the focus of your thesis.
Used skills Cloning, PCR, Gel electrophoresis, Transient expression, Protein pull-down, SDS-PAGE, Western blot, Growing plants, Proximity labeling, Data analysis, Scientific writing
Requirements Depending on your study program, a basic understanding of plant- and molecular-biology.