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Optimized barley phytase gene expression by focused FIND-IT screening for mutations in cis-acting regulatory elements
Claus Krogh Madsen1, Charles Alistair Brearley2, Jesper Harholt3
1Department of Agroecology, Aarhus University, Slagelse, Denmark.
Frontiers in Plant Science
|March 18, 2024
Summary
Researchers modified barley gene expression by altering cis-acting regulatory elements (CREs) in the phytase gene promoter. This improved grain quality for animal feed by enhancing phosphate availability and demonstrated a concept for modulating seed gene expression.
Area of Science:
- Plant molecular biology
- Agricultural science
- Biotechnology
Background:
- Cis-acting regulatory elements (CREs) control gene expression timing and strength in plants.
- Exploiting induced genetic variation in CREs for crop improvement is limited.
Purpose of the Study:
- To discover barley mutants with CRE variants in the promoter of the grain phytase (PAPhy_a) gene.
- To investigate the impact of altered PAPhy_a expression on grain quality and phosphate mobilization.
Main Methods:
- Digital PCR-based FIND-IT technology was used to identify barley mutants.
- Promoter CREs of the PAPhy_a gene were targeted for modification.
- Field trials and inositol phosphate profiling were conducted.
Main Results:
- Mutants with increased or decreased PAPhy_a gene expression and mature grain phytase activity (MGPA) were identified.
- Altered PAPhy_a expression did not affect agronomic performance but shortened phosphate mobilization lag time during germination.
- Increased MGPA enhances grain quality for animal feed by improving phosphate bioavailability.
Conclusions:
- Induced CRE variation in the PAPhy_a promoter offers a strategy to improve grain quality for animal feed.
- The study demonstrates a versatile concept for modulating gene expression in various seed-expressed genes, including storage proteins.
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