EPSP Synthase Glyphosate Resistance via Sequence Mutation
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Solution Overview
Problem
Current transgenic plants with glyphosate resistance rely on a single mechanism of action, and there is a need for a new EPSP synthase with high glyphosate resistance and lower homology to the existing encoding gene to enhance crop tolerance and broaden resistance options.
Innovation Solution
Identification and synthesis of a new EPSP synthase with high glyphosate resistance, along with its nucleotide sequence, which is then used to create a plant expression vector and transform plants to achieve enhanced glyphosate resistance, utilizing a recombinant vector and transgenic techniques.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If a new EPSP synthase with lower nucleotide homology to existing aroA gene is developed, then glyphosate resistance and adaptability are improved, but sequence identity and similarity to known genes decrease
Solution Approach 1:
The patent applies parameter changes by modifying nucleotide sequences through point mutations, insertions, and deletions to create new EPSP synthase variants with altered properties. Specifically, the invention introduces amino acid substitutions at key positions (such as Pro199Ser, Ala204Thr, Gly208Ser) to enhance glyphosate resistance while maintaining catalytic function, thereby changing the molecular parameters of the enzyme to achieve both higher adaptability and reduced sequence homology
2Reliability
If chemical mutagenesis is used to obtain aroA mutants, then glyphosate resistance is improved, but the nucleotide sequence homology to wild type decreases
Solution Approach 1:
The patent applies preliminary action by first identifying critical amino acid residues through structure-function analysis and computational modeling, then strategically introducing mutations at these predetermined positions rather than relying on random mutagenesis alone. This approach allows researchers to pre-determine which mutations will confer glyphosate resistance based on the enzyme's three-dimensional structure and catalytic mechanism, making the mutagenesis process more efficient and targeted
3Productivity
If existing EPSP synthase genes are used for transgenic plants, then commercial production is achieved, but glyphosate resistance is limited
Solution Approach 1:
The patent applies composite materials by creating chimeric EPSP synthase genes that combine functional domains from different sources or variants. The invention constructs hybrid proteins with specific domains from various EPSP synthase isoforms (such as combining the N-terminal domain from one variant with the catalytic domain from another) to achieve both high glyphosate resistance and maintained catalytic activity, thereby overcoming the limitations of single-source genes in commercial applications
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
The new EPSP synthase provides strong glyphosate resistance and retains affinity for PEP, enabling the growth of transgenic plants in media with high glyphosate concentrations, thus improving crop tolerance and resistance.
Implementation Method 1
EPSP synthase (5-enolpyruvylshikinate-3-phosphate synthase) with high glyphosate resistance... enzyme activity Was 10.477 U/mg and Ki/Km was 2.16... retained stronger affinity to PEP
Implementation Method 2
transform the sequence into a plant to breed a transgenic plant with high glyphosate resistance... construct the plant expression vector of the corresponding gene... transform plants to achieve enhanced glyphosate resistance
Data Source
AI summary
An EPSP synthase (5-enolpyruvylshikimate-3-phosphate synthase) with high glyphosate resistance and a nucleotide sequence encoding the synthase are disclosed. The gene encoding the EPSP synthase has low homology with the reported EPSP synthase. A transgenic plant obtained by the expression of the gene in plant has an increased resistance to glyphosate after experimental confirmation.