Virus Vector RNAi Silencing for Anthracnose Disease Control
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Solution Overview
Problem
Current control strategies for anthracnose disease in plants, caused by Colletotrichum spp., rely on expensive and environmentally harmful synthetic fungicides, and there is a need for a more effective and specific method to prevent infection without introducing transgenic plants.
Innovation Solution
A plant virus-based vector is modified to carry and express a partial sequence of the β-tubulin 2 gene of Colletotrichum spp., which, when ingested by the fungus, triggers RNA interference, silencing the fungal gene and preventing infection, using a transient RNAi system without causing viral symptoms in plants.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If synthetic fungicides are used to control anthracnose disease, then disease control effectiveness is improved, but environmental harm and cost increase
Solution Approach 1:
The patent replaces chemical fungicides with a biological mechanism (RNA interference) to control the disease. The VIGS vector delivers dsRNA that triggers RNAi in the fungus, substituting chemical intervention with a molecular biological approach that is environmentally friendly while maintaining disease control effectiveness
Solution Approach 2:
The patent introduces a plant virus-based VIGS vector as an intermediary to deliver the silencing sequence to the fungus. This vector acts as a mediator that carries the β-tubulin gene sequence into the fungal cells during infection, enabling specific gene silencing without direct chemical application
2Reliability
If transgenic plants are used to achieve disease resistance, then protection efficiency is improved, but biosafety concerns increase
Solution Approach 1:
The patent applies preliminary action by pre-introducing the VIGS vector containing the β-tubulin silencing sequence into the plant before fungal infection occurs. The plant is prepared with the silencing mechanism in advance, so when the fungus infects, the RNAi is immediately triggered, providing protection without making the plant transgenic
Solution Approach 2:
The patent uses a transient RNAi system where the VIGS vector is introduced temporarily to deliver the silencing sequence. The vector and dsRNA are short-lived and transient, achieving the protective effect without permanent genetic modification of the plant, thus avoiding transgenic biosafety issues
3Manufacturing precision
If a virus vector is introduced into plants to deliver silencing sequence, then gene silencing efficiency is improved, but viral symptom expression may occur
Solution Approach 1:
The patent extracts only the essential elements needed for silencing (the coat protein gene and movement protein gene sequences) from the viral genome, while removing or inactivating the pathogenic components. This allows the vector to deliver the silencing sequence efficiently without causing disease symptoms in the plant host
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 provides efficient and specific protection against anthracnose disease in olive plants and other hosts by replicating within the plant and activating the plant's defense mechanism to destroy the fungus, reducing the need for synthetic fungicides and ensuring biosafety.
Implementation Method 1
When a dsRNA sequence of a gene essential for the fungus survival is introduced into a plant through a virus vector, and the fungus 'feeds' on that plant, it ingests the plant cytoplasm and acquires the dsRNA; RNAi will be triggered, homologous RNAs will be targeted, the expression of that fungal gene will be prevented and fungus will be destroyed
Data Source
AI summary
The present invention discloses an isolated sequence having at least 95% homology with SEQ ID No 1, preferably at least 98% homology, even more preferably having at least 99% homology, the isolated sequence comprising at least one point mutation selected from: adenine to thymine at nucleotide 3200, cytosine to guanine at nucleotide 3361, guanine to cytosine at nucleotide 3365 and/or guanine to thymine at nucleotide 3368. The invention also relates to a process for silencing a target gene in a plant comprising the steps of i) obtaining a construct according to the previous claim; ii) introducing the construct inside the plant, preferably by using viral suspensions; inoculating into plants, preferably by spraying plants, even more preferably by spraying plants with a compressor set comprising an airbrush; iii) activating the plant's specific defence mechanism, consequently silencing the target gene. The present invention also discloses a composition for plant disease management and/or plant immunization.