Viral Meristem Modulation for Targeted Plant Gene Modification
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Solution Overview
Problem
Crop plants like cotton are recalcitrant to transformation and regeneration, making traditional genome editing inefficient and labor-intensive for targeted genetic modification, particularly in meristems which resist genetic manipulation, hindering the incorporation of desired traits such as pest resistance and improved growth.
Innovation Solution
A method involving silencing a meristem maintenance gene to induce a phase change from indeterminate to determinate, combined with genetic manipulation of a second gene, using viral vectors with siRNA targeting the meristem maintenance gene and engineered RNA sequences to modify the plant meristem, facilitating detectable new phenotypes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional genome editing methods are used in recalcitrant crop plants like cotton, then transformation and regeneration can be attempted, but the process becomes inefficient, time-consuming, and labor-intensive
Solution Approach 1:
The patent segments the genetic modification process into two distinct phases: (1) silencing the meristem maintenance gene to induce phase change from indeterminate to determinate state, and (2) introducing the desired trait gene. This segmentation allows each phase to be optimized independently, with the meristem phase change occurring naturally without requiring tissue culture regeneration, thereby eliminating the time-consuming regeneration step while maintaining genetic modification efficiency
Solution Approach 2:
The patent applies preliminary action by first silencing the meristem maintenance gene (such as WUSCHEL or CLAVATA3) to induce a phase change in the meristem from indeterminate to determinate state before introducing the desired trait. This preliminary meristem modulation creates a permissive state that facilitates subsequent genetic manipulation and direct regeneration, eliminating the need for lengthy tissue culture procedures
2Ease of manufacture
If viral vectors are used to introduce genetic material into meristems, then transformation efficiency improves, but meristems resist genetic manipulation due to poorly understood defenses
Solution Approach 1:
The patent applies preliminary action by first silencing the meristem maintenance gene to induce a phase change from indeterminate to determinate state, which creates a permissive state that facilitates subsequent viral-based genetic manipulation. This preliminary meristem modulation occurs before introducing the viral vector containing the desired trait, thereby reducing the meristem's defensive resistance and enabling efficient transformation without requiring lengthy tissue culture
Solution Approach 2:
The patent changes the physiological state parameter of the meristem by inducing a phase change from indeterminate to determinate through silencing of meristem maintenance genes. This parameter change in meristem development state fundamentally alters the meristem's properties, making it more susceptible to viral-based genetic manipulation and reducing its resistance to foreign nucleic acids
3Ease of operation
If meristem maintenance genes are silenced to induce phase change, then access to germline cells improves, but the meristem's natural defenses against foreign nucleic acids are triggered
Solution Approach 1:
The patent applies preliminary action by first silencing the meristem maintenance gene to induce phase change and create a permissive state, which then facilitates the introduction of viral vectors containing desired traits. The sequence is carefully ordered: meristem gene silencing occurs first to modify the meristem state, followed by viral vector introduction, thereby ensuring that the meristem is more receptive to foreign nucleic acids at the time of introduction
Solution Approach 2:
The patent skips the traditional intermediate step of tissue culture regeneration by directly transitioning from meristem phase change to viral-based genetic manipulation. This rushing through the process eliminates the time-consuming tissue culture step while maintaining efficient access to germline cells, as the phase-changed meristem directly gives rise to edited germline cells in flowers and ultimately edited seeds
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 stabilizes targeted genetic modifications in crop plants by making meristems more susceptible to viral-based manipulation, enabling efficient introduction of traits like pest resistance and improved growth without lengthy tissue culture.
Implementation Method 1
silencing a meristem maintenance gene in the plant to induce a phase change in the plant meristem from an indeterminate to a determinate state
Implementation Method 2
genetic manipulation of a second gene in the plant, wherein genetic manipulation of the second gene in combination with silencing the meristem maintenance gene results in a detectable new phenotype
Data Source
AI summary
The present disclosure includes methods of producing genetically modified plants with a detectable new phenotype via combined silencing of a meristem maintenance gene and genetic manipulation of a second gene. Also included are genetically modified plants having engineered genetic modifications including a silenced meristem maintenance gene and a genetic modification of a second gene and systems for co-modifying meristem maintenance and other genes in a plant.