Plant Immunity via Pathogen sRNA Neutralization
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Solution Overview
Problem
Botrytis cinerea, a fungal pathogen, suppresses plant immunity by using small RNAs (sRNAs) that hijack the host's gene silencing mechanisms, leading to increased susceptibility of crops to infection, resulting in significant annual losses worldwide.
Innovation Solution
Introduction of a heterologous expression cassette in plants with a promoter operably linked to a polynucleotide that is complementary to or mediates destruction of pathogen sRNAs, or encodes an sRNA-resistant target protein, to enhance pathogen resistance. This includes using short tandem target mimics (STTMs) or antisense nucleic acids to block pathogen sRNAs and expressing sRNA-resistant target genes involved in plant immunity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If pathogen sRNAs are allowed to function in the host plant, then the pathogen can suppress host immunity and establish infection, but the plant becomes highly susceptible to disease and suffers crop losses
Solution Approach 1:
The patent applies preliminary anti-action by introducing polynucleotides into the plant that are complementary to pathogen sRNAs before the pathogen can establish infection. These polynucleotides preemptively bind to and neutralize pathogen sRNAs, preventing them from suppressing host immunity genes. This anticipatory defense mechanism blocks the pathogen's immune suppression strategy before it can take effect, thereby reducing plant susceptibility while allowing normal pathogen interaction processes to occur.
2Object-affected harmful factors
If the plant's natural gene silencing mechanism is used against the pathogen, then pathogen sRNAs can be targeted and destroyed, but this requires introducing heterologous expression cassettes and polynucleotides into the plant
Solution Approach 1:
The patent applies self-service by harnessing the plant's own endogenous RNA interference (RNAi) machinery to combat pathogen sRNAs. The introduced polynucleotides serve as triggers that activate the plant's natural gene silencing pathways, including Dicer-like enzymes and Argonaute proteins, which then process and degrade the pathogen sRNAs. This approach leverages the plant's existing molecular defense infrastructure rather than requiring entirely new systems, thereby reducing the complexity of the genetic modification while effectively neutralizing pathogen sRNA activity.
3Reliability
If polynucleotides complementary to pathogen sRNAs are introduced, then pathogen sRNA function is blocked and plant immunity is enhanced, but the plant requires genetic modification with heterologous expression cassettes
Solution Approach 1:
The patent applies universality by designing polynucleotides that can target multiple pathogen sRNA sequences with a single genetic construct. The expression cassettes are engineered to produce polynucleotides with broad specificity that can simultaneously bind to and neutralize various pathogen sRNAs involved in immune suppression. This multi-functional approach enhances plant immunity against multiple sRNA targets while minimizing the number of genetic modifications required, thereby improving ease of manufacture compared to introducing separate constructs for each sRNA target.
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 approach significantly reduces the susceptibility of plants to Botrytis cinerea infection by blocking the pathogen's ability to suppress host immunity, leading to enhanced disease resistance and reduced fungal biomass, thereby mitigating the economic impact of crop losses.
Implementation Method 1
a polynucleotide that is complementary to, or mediates destruction, of a plant immunity suppressing sRNA of a pathogen
Implementation Method 2
sRNAs induce gene silencing by binding to Argonaute (AGO) proteins and directing the RNA-induced silencing complex (RISC) to genes with complementary sequences
Data Source
AI summary
The present invention relates to pathogen-resistant plants comprising a heterologous expression cassette, the expression cassette comprising a promoter operably linked to a polynucleotide that is complementary to, or mediates destruction, of a plant immunity suppressing sRNA of a pathogen, wherein the plant is less susceptible to the pathogen compared to a control plant lacking the expression cassette. Methods of making and cultivating pathogen-resistant plants are also provided.