uORF Polynucleotides Regulate GGP Expression and Ascorbate
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Solution Overview
Problem
There is a significant variability in ascorbate content in non-green edible plant tissues, and the mechanisms regulating ascorbate biosynthesis in plants are not well understood, limiting the ability to manipulate ascorbate production effectively.
Innovation Solution
The use of specific polynucleotides encoding peptides or their variants, which are operably linked to the GDP-L-Galactose phosphorylase (GGP) gene, to modulate GGP activity and ascorbate content in plants, allowing for genetic manipulation and regulation of ascorbate production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of information
If the L-Galactose pathway genes are identified and characterized, then the understanding of ascorbate biosynthesis is improved, but the ability to effectively manipulate ascorbate production is limited due to unknown regulatory mechanisms
Solution Approach 1:
The patent performs preliminary identification and characterization of the L-Galactose pathway genes and their regulatory elements (promoters, enhancers, transcription factor binding sites) before attempting manipulation. This preliminary genetic mapping enables subsequent targeted manipulation of ascorbate production by providing the necessary knowledge base of regulatory mechanisms.
2Quantity of substance
If ascorbate content in plant tissues is increased, then the nutritional value and antioxidant capacity are improved, but the variability in ascorbate content across different plant tissues remains unexplained and difficult to control
Solution Approach 1:
The patent identifies feedback regulatory mechanisms in the L-Galactose pathway, where ascorbate levels and related metabolites regulate gene expression through transcription factors and epigenetic modifications. Understanding these feedback loops enables controlled manipulation of ascorbate content by targeting specific regulatory nodes in the pathway.
Solution Approach 2:
The patent employs parameter changes in gene expression levels, promoter strength, and transcription factor activity to control ascorbate content. By modifying these parameters through genetic engineering, the variability in ascorbate content across different plant tissues can be standardized and controlled.
3Stability of the object's composition
If the variability in ascorbate content among different plant species and cultivars is reduced, then consistency in nutritional quality is improved, but the genetic mechanisms underlying this variability are not well understood
Solution Approach 1:
The patent segments the ascorbate biosynthesis pathway into distinct genetic and regulatory components (GGP gene, promoters, enhancers, transcription factors). This segmentation allows systematic analysis of each component's contribution to ascorbate content variability, enabling targeted manipulation to achieve consistency across different plant varieties.
Data Source
AI summary
The invention provides isolated polynucleotides comprising sequences encoding a u ORF peptides and variants and fragments thereof. The invention also provides constructs and vectors containing the polynucleotides. The invention further provides cells, plant cells and plants transformed with the polynucleotides and constructs. The invention also provides methods of using the polynucleotides to control expression of operably linked polynucleotides. The invention also provides methods of manipulating GDP-L-Galactose phosphorylase (GGP) expression and ascorbate production in plants utilizing the polynucleotides of the invention.