Københavns Universitet

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MSc-project Gene editing towards increased yield and photosynthetic efficiency in Potato at Københavns Universitet

Project description:

Current breeding strategies are limited in their capabilities to push the yield barrier while our society is facing dramatic changes in climate and population structure. By 2050, the primary agricultural productivity must be doubled to meet the anticipated requirements to feed population and livestock. The challenge will be to achieve this expansion sustainably while maintaining food quality and reducing the environmental impact of current resource intensive agricultural practices.

Enhancing photosynthetic efficiency in a targeted fashion towards increased plant growth and biomass production is the main goal of the ReTraQue-Project here at PLEN. This is tackled in several important crop species like: Alfalfa, barley, ryegrass, wheat and potato. We target regulatory genes in the photosynthetic network to alter their expression profile and thereby modulate the crops towards increases in biomass and yield.

Perspectives:

As part of an ongoing project, you will be involved in the phenotypic validation, targeted genome sequencing and genome editing of two potato cultivars. Based on previous greenhouse and field trials for photosynthetic efficiency at the site of our collaborator DANESPO, we selected two cultivars for detailed photosynthetic evaluation under controlled conditions. Further, both cultivars will be gene edited in order to modulate the photosynthetic gene regulatory network, aiming towards improved tuber yield. This project spans from target gene selection and identification over gene editing towards mutant line evaluation including the following methods.

  • Photosynthetic evaluation of the selected potato cultivars under control and stress conditions (fluctuating light).
    • Imaging PAM, Dual PAM and OJIP
    • Western Blot, RT-qPCR
  • Determination of target genes according the data obtained.
    • Comparison to already exiting greenhouse and field trail data.
  • Target sequence identification
    • Targeted DNA sequencing by pulldown enrichment (MinION, Oxford Nanopore Technologies)
  • Guide RNA design and cloning
  • Cell culture and transformation (Agrobacterium mediated transformation on potato callus)
    • Including the transformation of already existing control constructs
  • Regeneration of edited plantlets and genotyping
    • Genotyping PCR (targeted amplicon sequencing)
  • Evaluation of candidate lines.
    • Photosynthetic efficiency (Imaging PAM, Dual PAM and OJIP)
    • Network response (RT-qPCR, Western Blot)
    • Yield parameter (Tuber size and quality measurement in cooperation with DANESPO)
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