Rice Yield Improvement via PYL Gene Knockout

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Solution Overview

Problem

Current methods for improving rice yield are limited by the growth inhibition associated with increased stress resistance induced by Abscisic Acid (ABA), which is mediated by the PYL family of ABA receptor genes, with little research on specific gene functions in rice and no reported rice PYL mutants.

Innovation Solution

The method involves genetically engineering rice plants to knock out multiple members of the PYL gene family using CRISPR/Cas9 technology, reducing their expression or activity to enhance traits such as increased yield, biomass, and reduced seed dormancy without significant delays in the heading stage.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If ABA content is increased to enhance stress resistance, then plant resistance to stress is improved, but plant growth is inhibited

Engineering Contradiction:

Improvestress resistanceVSAvoidplant growth

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The invention segments the PYL gene family into multiple individual gene targets (PYL1, PYL4, PYL6, etc.) and selectively knocks out specific members rather than treating ABA signaling as a unified pathway. This allows differential modulation of stress resistance and growth functions that are mediated by different PYL genes

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The invention applies local quality by creating non-uniform modification across the PYL gene family - specifically knocking out PYL1, PYL4, and PYL6 while leaving other PYL genes intact. This selective approach allows different regions of the gene family to have different functional outcomes, achieving growth promotion without completely abolishing stress resistance

Inventive Principle:

Principle #3Local quality

2Productivity

If multiple PYL genes are knocked out to promote growth, then yield and biomass are increased, but seed dormancy is reduced

Engineering Contradiction:

ImproveyieldVSAvoidseed dormancy

Core Design Contradiction:

ProductivityVSStability of the object's composition

Solution Approach 1:

The invention applies partial action by knocking out only three specific PYL genes (PYL1, PYL4, PYL6) out of the entire PYL gene family, rather than knocking out all PYL genes. This partial modification is sufficient to promote growth and increase yield while maintaining some level of seed dormancy control through the remaining intact PYL genes

Inventive Principle:

Principle #16Partial or excessive action

3Productivity

If CRISPR/Cas9 is used to edit multiple PYL genes simultaneously, then gene editing efficiency is improved, but off-target effects and plant complexity increase

Engineering Contradiction:

Improvegene editing efficiencyVSAvoidplant complexity

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The invention segments the gene editing task into multiple independent sgRNA targets within a single CRISPR/Cas9 system. By designing specific guide RNAs for PYL1, PYL4, and PYL6, the system can simultaneously edit multiple genes without requiring multiple transformation events, thus improving efficiency while managing complexity

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The invention applies universality by using a single CRISPR/Cas9 expression system to perform multiple gene editing functions simultaneously. The same Cas9 nuclease and selection marker system is used to target and edit multiple different PYL genes, reducing the overall complexity compared to using separate editing systems for each gene

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 significantly promotes rice growth and increases yield, with simultaneous knockout of PYL1, PYL4, and PYL6 showing the best growth status and agronomic traits, resulting in a 25-31% increase in grain yield compared to wild-type rice.

Implementation Method 1

Cas9 nuclease is guided by short sgRNA (single guide RNA) to cut the DNA sequence complementary to the sgRNA recognition region

Methodology Applied

Scientific EffectCRISPR/Cas9 gene editing:

Implementation Method 2

The binding of ABA to PYL causes the conformational change of the protein, which causes and promotes the binding of PYL to PP2C (clade A type 2C protein phosphatase) protein. The formation of ABA-PYL-PP2C complex inhibits the activity of PP2C

Methodology Applied

Scientific EffectProtein-protein binding:

Data Source

PatentUS11603537B2Method for improving rice yield by jointly knocking out ABA receptor PYL family genes and use thereof

Publication Date: 2023.03.14 CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI

AI summary

Provided are a method for improving rice yield by jointly knocking out ABA receptor PYL family genes and a use thereof.