Alpha-WOLF 26 Allele Resolves Spinach Downy Mildew Resistance

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Solution Overview

Problem

Current spinach varieties are susceptible to multiple races of Peronospora farinosa f. sp. spinaciae, leading to downy mildew, which threatens spinach productivity as the pathogen continuously evolves to break existing resistances.

Innovation Solution

A new allelic variant of the Alpha-WOLF gene, designated alpha-WOLF 26, confers resistance to at least downy mildew races Pfs:15, Pfs:16, and Pfs:17, characterized by specific amino acid motifs and sequence identity in the CC-NBS-LRR protein, providing a molecular basis for resistance.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If existing resistance genes are used in spinach varieties, then resistance to certain Peronospora races is achieved, but the pathogen continuously evolves new races that break these resistances

Engineering Contradiction:
Improvedurability of resistanceVSAvoidbreadth of resistance against multiple races
Core Design Contradiction:
ReliabilityVSAdaptability or versatility

Solution Approach 1:

The patent applies dynamics by creating a stacks of multiple R-genes (DMR1, DMR6, p10, and p25) that can dynamically respond to different Peronospora races. This stack configuration allows the plant to maintain resistance as the pathogen evolves, since multiple resistance mechanisms work together rather than relying on a single static resistance gene that can be easily broken by pathogen adaptation.

Inventive Principle:
Principle #15Dynamics

Solution Approach 2:

The patent employs composite materials by combining multiple resistance genes (DMR1, DMR6, p10, p25) into a stacked configuration. This composite genetic structure provides broader and more durable resistance against multiple Peronospora races compared to individual resistance genes, as the combined effect of multiple R-genes creates a more robust defense system that is harder for the pathogen to overcome.

Inventive Principle:
Principle #40Composite materials

2Adaptability or versatility

If multiple resistance genes are stacked to broaden resistance spectrum, then resistance against more races is achieved, but the genetic complexity and breeding difficulty increase

Engineering Contradiction:
Improvebreadth of resistance against multiple racesVSAvoidgenetic stack complexity
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent applies universality by designing a resistance stack where multiple R-genes (DMR1, DMR6, p10, p25) work together to provide broad-spectrum resistance against multiple Peronospora races. This multi-functional genetic stack can simultaneously defend against different pathogen races, making it a universal solution for downy mildew resistance in spinach rather than requiring separate resistance genes for each specific race.

Inventive Principle:
Principle #6Universality (Multi-functionality)

3Productivity

If conventional breeding methods are used to develop resistant varieties, then resistance can be introduced, but the process is slow and cannot keep pace with rapidly evolving pathogen races

Engineering Contradiction:
Improvespeed of resistance developmentVSAvoideffectiveness against new races
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent applies preliminary action by pre-characterizing and validating the resistance profiles of multiple R-genes (DMR1, DMR6, p10, p25) against different Peronospora races before deployment. This advance preparation allows breeders to quickly assemble effective resistance stacks without needing to conduct extensive field trials for each new pathogen race, thereby accelerating the development of resistant varieties while maintaining reliability against emerging threats.

Inventive Principle:
Principle #10Preliminary action

Data Source

PatentUS11820993B2<i>Peronospora </i>resistance in <i>Spinacia oleracea </i>
Publication Date: 2023.11.21 RIJK ZWAAN ZAADTEELT & ZAADHANDEL BV

AI summary

The present invention relates to an allele designated alpha-WOLF 26 which confers resistance to at least one Peronospora farinosa f. sp. spinacea race, wherein the protein encoded by said allele is a CC-NBS-LRR protein that may comprise in its amino acid sequence: a) the motif “MAEIGYSVC” SEQ ID NO: 1 at its N-terminus; and b) the motif “KWMCLR” SEQ ID NO: 2; and wherein the LRR domain of the protein has in order of increased preference at least 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 100% sequence identity to SEQ ID NO: 10. The allele when present in a spinach plant confers complete resistance to at least Peronospora farinosa f. sp. spinacea race Pfs:7, Pfs:10, Pfs:11, Pfs:12, Pfs:13, Pfs:14, Pfs:15, Pfs:16, Pfs:17, and does not confer resistance to downy mildew race Pfs:8 and Pfs:9.