Soybean ROD1 Gene Modulation for Oleic Acid Control

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Solution Overview

Problem

Current methods for modifying fatty acid composition in soybeans to increase unsaturated fatty acids, such as oleic acid, are limited by the complexity of the ROD1 gene sequences and the difficulty in identifying functional ROD1 genes in soybeans, particularly due to polyploid history, which complicates the isolation of mutant alleles.

Innovation Solution

The introduction of knock-out ROD1 genes or inhibitory RNAs targeting ROD1 genes in soybeans to modulate the expression of these genes, thereby increasing the levels of oleic acid (C18:1) in soybean seed oil, using chimeric genes with plant-expressible promoters and transcription termination regions, and employing mutagenic treatments to identify plants with increased C18:1 levels.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If FAD2 gene silencing is used to increase oleic acid levels, then C18:1 content increases, but membrane unsaturation is reduced and plant growth is compromised

Engineering Contradiction:
Improveoleic acid contentVSAvoidplant growth
Core Design Contradiction:
Quantity of substanceVSReliability

Solution Approach 1:

The invention targets ROD1 gene specifically in seed tissue to modify fatty acid composition, leaving other tissues unaffected. This spatially localized approach allows high oleic acid content in seeds while maintaining normal membrane unsaturation and plant growth in vegetative tissues.

Inventive Principle:
Principle #3Local quality

Solution Approach 2:

The invention uses ROD1 gene as an intermediary target instead of directly targeting FAD2. ROD1 acts as a regulatory gene that controls FAD2 expression specifically in seeds, allowing indirect modification of fatty acid composition without directly disrupting FAD2 function in other tissues.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Device complexity

If multiple ROD1 gene copies are present due to polyploidy, then gene complexity increases, but identification of functional genes becomes difficult

Engineering Contradiction:
Improvegene sequence complexityVSAvoidfunctional gene identification
Core Design Contradiction:
Device complexityVSDifficulty of detecting and measuring

Solution Approach 1:

The invention isolates and characterizes individual ROD1 gene copies (ROD1-1 and ROD1-2) separately, determining their specific sequences, expression patterns, and functional contributions. This segmentation allows clear identification of which copies are functional and how they contribute to fatty acid composition.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The invention uses molecular markers and expression analysis to feedback on the functional status of each ROD1 copy, enabling identification of functional genes among multiple copies through systematic characterization of sequence variations and expression patterns.

Inventive Principle:
Principle #23Feedback

3Quantity of substance

If partial downregulation of FAD2 is used, then oil composition is modified, but trans fatty acids are still produced and nutritional value is limited

Engineering Contradiction:
Improvefatty acid compositionVSAvoidtrans fatty acid production
Core Design Contradiction:
Quantity of substanceVSObject-affected harmful factors

Solution Approach 1:

The invention creates transgenic copies of ROD1 gene with modified expression characteristics (e.g., ROD1-1 and ROD1-2 with different expression patterns). These engineered copies allow precise control over FAD2 regulation to achieve complete elimination of trans fatty acids while maintaining high oleic acid content.

Inventive Principle:
Principle #26Copying

Data Source

PatentUS9968041B2Soybean ROD1 gene sequences and uses thereof
Publication Date: 2018.05.15 BASF AGRICULTURAL SOLUTIONS SEED US LLC

AI summary

The present invention relates to soybean ROD1 nucleic acid sequences and proteins and the use thereof to create plants with increased levels of C18:1 and reduced levels of saturated fatty acids in the seeds.