Soybean Transformation via External Herbicide Application
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Solution Overview
Problem
Current methods for soybean transformation using ALS inhibitors in culture media result in high false-positive rates and low transformation efficiency, posing challenges for large-scale genetic transformation and breeding with herbicide tolerance traits.
Innovation Solution
A method involving the external application of sulfonylurea herbicide hydrolase as a selective marker, using a recombinant vector with a gene encoding sulfonylurea herbicide hydrolase, and applying ALS inhibitors like tribenuron methyl to select transformed soybean cells through an Agrobacterium-mediated process.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If ALS inhibitors are disposed in culture media for selecting transformed plants, then herbicide resistance traits can be selected, but false-positive proportion increases and transformation efficiency decreases
Solution Approach 1:
The patent introduces a sulfonylurea herbicide hydrolase gene as an intermediary enzyme that degrades the selective agent (sulfonylurea herbicide) into non-toxic products. This mediator allows transformed cells to survive herbicide treatment while preventing false positives, thereby resolving the contradiction between selection accuracy and transformation efficiency
Solution Approach 2:
The patent changes the chemical parameter of the selective agent by using sulfonylurea herbicide hydrolase to convert the toxic herbicide into non-toxic degradation products. This parameter change enables differentiated survival between transformed and non-transformed cells, improving both selection accuracy and transformation efficiency
2Reliability
If antibiotic resistance genes are used as selective markers, then transformation selection is effective, but biosafety concerns arise regarding ecological environment and food safety
Solution Approach 1:
The patent converts the potentially harmful antibiotic resistance genes into beneficial herbicide degradation enzymes. The sulfonylurea herbicide hydrolase gene provides selection capability while degrading herbicides into safe products, eliminating biosafety risks associated with antibiotic resistance and providing ecologically friendly alternative markers
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach significantly improves transformation efficiency and positive plant rates, optimizing the screening system and reducing costs, while enabling the development of transgenic soybeans with high tolerance to sulfonylurea herbicides.
Implementation Method 1
a gene encoding a sulfonylurea herbicide hydrolase
Data Source
AI summary
The present invention relates to a method of improving soybean transformation efficiency, which comprises: transforming a plant cell using a recombinant vector containing a gene of interest and a gene encoding a sulfonylurea herbicide hydrolase; screening and culturing the transformed plant cell by external application of an ALS inhibitor, using the gene encoding the sulfonylurea herbicide hydrolase as a selective marker; selecting a plant cell that has not been killed and/or not been inhibited. The present invention firstly proposes that a selective agent is added to a proliferation medium and a differentiation medium in a manner of external application during plant transformation process, and optimizes the effective screening concentration range of the selective agent, so the transformation efficiency is remarkably increased, and the proportion of positive plants obtained in the progeny thereof is significantly increased; at the same time, the transgenic plants obtained by the transformation using the sulfonylurea herbicide hydrolase gene as a selective marker in the present invention have high commercial value, good resistance and genetic stability.