dsRNA Plant Expression for Nematode Gene Silencing

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Solution Overview

Problem

Current methods for controlling root knot nematodes are either non-selective, costly, or have limited effectiveness, as they disrupt beneficial organisms and are not durable, leading to the need for an efficient method to reduce nematode multiplication and reproduction across multiple stages.

Innovation Solution

The method involves transforming host plant cells with nucleic acids to produce double-stranded RNA that inhibits the expression of the FMRFamide-like peptide 14 gene in root knot nematodes, using a RNAi vector construct to silence the flp-14 gene, thereby reducing nematode multiplication and reproduction.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If chemical agents are used to control root-knot nematodes, then nematode control effectiveness is improved, but harmful effects on beneficial microorganisms and environmental persistence increase

Engineering Contradiction:

Improvenematode control effectivenessVSAvoidharmful effects on beneficial microorganisms

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent uses RNA molecules as intermediaries to transfer the control function from the plant to the nematode. The plant produces dsRNA that is taken up by the nematode during feeding, which then triggers RNAi to silence essential nematode genes. This intermediary mechanism allows selective control of nematodes without affecting beneficial microorganisms, as the RNAi mechanism is species-specific.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent changes the control mechanism from chemical toxicity to genetic silencing. By targeting specific gene expression in nematodes through dsRNA-mediated RNAi, the control method achieves high specificity. The parameter change from non-selective chemical action to selective genetic targeting eliminates harm to beneficial organisms while maintaining effectiveness against nematodes.

Inventive Principle:

Principle #35Parameter changes

2Reliability

If host plant resistance genes are used to control nematodes, then nematode resistance is improved, but durability decreases due to resistance-breaking populations

Engineering Contradiction:

Improvenematode resistanceVSAvoiddurability of resistance

Core Design Contradiction:

ReliabilityVSDuration of action of stationary object

Solution Approach 1:

The patent changes the resistance mechanism from broad-spectrum genetic resistance to highly specific gene silencing. By targeting essential nematode genes involved in development and reproduction through RNAi, the resistance becomes more durable. The specificity of RNAi to particular gene sequences makes it difficult for nematodes to develop resistance through mutation, as any mutation in the target gene would likely be lethal to the nematode.

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent establishes continuous gene silencing action throughout the nematode lifecycle. The dsRNA is continuously produced in the plant and remains active during nematode feeding, ensuring sustained suppression of nematode development and reproduction. This continuous action prevents the intermittent exposure that allows resistance-breaking populations to emerge.

Inventive Principle:

Principle #20Continuity of useful action

3Productivity

If RNAi is used to silence nematode genes, then nematode multiplication is reduced, but specificity and effectiveness against multiple lifecycle stages must be maintained

Engineering Contradiction:

Improvereduction in nematode multiplicationVSAvoideffectiveness across multiple lifecycle stages

Core Design Contradiction:

ProductivityVSAdaptability or versatility

Solution Approach 1:

The patent uses a universal RNAi mechanism that functions across all nematode lifecycle stages. The dsRNA targets essential genes that are expressed throughout development, from early juvenile stages through reproduction. This universal approach allows a single intervention to affect multiple lifecycle stages, achieving both high productivity in reducing multiplication and versatility across developmental stages.

Inventive Principle:

Principle #6Universality (Multi-functionality)

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 results in a significant reduction of approximately 78% in the nematode multiplication index, along with decreases in female population, egg production, and gall formation, providing a durable and effective control mechanism.

Implementation Method 1

double-stranded RNA that inhibits the expression of the FMRFamide-like peptide 14 gene in root knot nematodes, using a RNAi vector construct to silence the flp-14 gene

Methodology Applied

Scientific EffectRNA interference (RNAi):

Data Source

PatentUS10308954B2Method for the control of nematodes in plants

Publication Date: 2019.06.04 ICAR

AI summary

The present invention relates to the field of double-stranded RNA (ds RNA) mediated gene silencing of root knot nematodes. The invention particularly provides an effective method for reducing the number of reproducing population and the number of progenies per individual of the root knot nematodes. The present invention also relates to host delivered dsRNA for controlling infection of root knot nematodes.