FLS3 Protein Engineering for Broad Bacterial Pathogen Detection
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Solution Overview
Problem
Current methods lack effective identification and utilization of plant-specific pattern recognition receptors (PRRs) for broadening disease resistance in crops, as few cloned PRR genes are available, and progress in genetic control of pathogen resistance is limited.
Innovation Solution
Development of a nucleic acid construct encoding the FLAGELLIN-SENSING 3 (FLS3) protein, including a 5′ heterologous DNA promoter and 3′ terminator sequence, to enhance disease resistance in plants by expressing the FLS3 protein or applying the flgII-28 peptide, thereby recognizing a broader range of bacterial pathogens.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If FLS2 is used to detect flagellin, then detection of bacterial pathogens is achieved, but pathogens with divergent flg22 region can evade detection
Solution Approach 1:
The patent introduces a second PRR (FLS3) that recognizes a different epitope (flgII-28) on flagellin, creating a multi-functional detection system. This allows the plant to recognize both flg22 and flgII-28 containing pathogens, broadening the recognition range while maintaining reliable detection of specific bacterial patterns.
Solution Approach 2:
The patent changes the detection parameter from recognizing only flg22 epitope to also recognizing flgII-28 epitope through FLS3 expression. This parameter expansion allows detection of pathogens that have evolved to evade FLS2 by modifying their flg22 region, while FLS3 continues to detect them through the conserved flgII-28 region.
2Adaptability or versatility
If species-specific PRRs are transferred to broaden disease resistance, then pathogen recognition is enhanced, but few cloned PRR genes are available for transfer
Solution Approach 1:
The patent performs preliminary identification and characterization of FLS3 from tomato before transfer to other species. By having the FLS3 gene sequence and functional understanding ready in advance, the patent enables immediate transfer and expression in non-Solanaceous plants, overcoming the limitation of unavailable cloned PRR genes.
Solution Approach 2:
The patent uses FLS3 as an intermediary receptor that mediates recognition of flgII-28 in plants that lack this natural capability. By introducing FLS3 into Arabidopsis and other non-Solanaceous plants, it acts as a bridge to confer new recognition specificity for bacterial pathogens.
3Adaptability or versatility
If FLS3 is expressed in non-Solanaceous plants, then recognition of flgII-28 is conferred, but genetic engineering complexity increases
Solution Approach 1:
The patent segments the FLS3 gene into manageable components (coding sequence, promoter, terminator) that can be assembled into an expression construct. This segmentation simplifies the genetic engineering process by allowing modular assembly of the transgene, making it easier to introduce FLS3 into diverse plant species despite the increased complexity of cross-species expression.
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 expression of FLS3 protein in plants confers increased resistance to bacterial infections, allowing for enhanced immune responses and improved disease resistance, leveraging natural variation to combat pathogens that evade detection by conventional mechanisms.
Implementation Method 1
the FLS3 protein, which binds a flagellin-derived MAMP that involves a peptide region known as flgII-28 thereby activating innate immunity
Data Source
AI summary
One aspect of the present invention relates to a nucleic acid construct that includes a nucleic acid molecule that encodes FLAGELLIN-SENSING 3 (“FLS3”) protein; a 5′ heterologous DNA promoter sequence; and a 3′ terminator sequence, where the nucleic acid molecule, the DNA promoter sequence, and the terminator sequence are operatively coupled to permit transcription of the nucleic acid molecule. The present invention also relates to a method of imparting disease resistance to a plant. This method involves transforming a plant or a plant seed with a nucleic acid molecule that increases expression of an FLS3 protein, where said transforming is effective in imparting disease resistance to the transformed plant or to a transgenic plant produced from the transformed plant seed. The present invention also relates to methods of expressing a nucleic acid molecule in a plant, identifying a candidate plant suitable for breeding that displays enhanced disease resistance, and enhancing efficiency of transformation of a plant by Agrobacterium.