Spinach WOLF Allele Stacking for Broad Downy Mildew Resistance

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Solution Overview

Problem

Current spinach cultivars lack resistance to all known pathogenic races of Peronospora farinosa f. sp. spinaciae, and new races continuously emerge, posing a threat to the spinach industry, necessitating the development of broad-spectrum resistance genes.

Innovation Solution

Identification and utilization of WOLF genes, specifically alpha- and beta-WOLF genes, and their alleles, which encode CC-NBS-LRR proteins, to modify the resistance profile of spinach plants by introducing or modifying these genes through traditional breeding or genetic engineering, enabling tailored resistance to multiple pathogenic races.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If existing resistance genes are used in spinach cultivars, then resistance to downy mildew is provided initially, but the resistance is overcome by the fungus over time as new pathogenic races emerge

Engineering Contradiction:
Improveresistance durabilityVSAvoidresistance breadth
Core Design Contradiction:
ReliabilityVSAdaptability or versatility

Solution Approach 1:

The patent combines multiple WOLF alleles (alpha-WOLF and beta-WOLF variants) into single spinach cultivars to create stacked resistance profiles. This merging of different resistance genes allows the plant to resist multiple pathogenic races simultaneously, preventing the fungus from overcoming resistance through single-race adaptation.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The WOLF gene family serves multiple resistance functions against different pathogenic races of Peronospora farinosa. By identifying and deploying universal resistance mechanisms that recognize conserved fungal elements, the cultivars achieve broad-spectrum resistance that remains effective across evolving pathogen populations.

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

2Measurement precision

If phenotypic assays are used to screen for resistance, then resistance can be detected, but the process is time-consuming and requires screening many accessions

Engineering Contradiction:
Improveresistance detection accuracyVSAvoidbreeding cycle duration
Core Design Contradiction:
Measurement precisionVSLoss of time

Solution Approach 1:

The patent performs preliminary genotypic screening using molecular markers linked to WOLF alleles before conducting phenotypic resistance assays. This preliminary identification of resistance-associated genes allows breeders to select promising accessions early in the breeding process, reducing the number of time-consuming phenotypic screenings required and accelerating the overall breeding cycle.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent replaces labor-intensive manual phenotypic screening with molecular marker-based genotypic detection. By using DNA markers associated with WOLF alleles, the system can rapidly identify resistance traits without requiring extensive growing and visual assessment, significantly reducing time and resource requirements.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Data Source

PatentEP3410845B1Method for modifying the resistance profile of spinacia oleracea to downy mildew
Publication Date: 2026.04.29 RIJK ZWAAN ZAADTEELT & ZAADHANDEL BV
  • EP3410845B1 patent drawingFigure 1
  • EP3410845B1 patent drawingFigure 2
  • EP3410845B1 patent drawing

AI summary

The present invention relates to a method for modifying the resistance profile of a spinach plant to Peronospora farinosa f. sp. spinaciae, comprising introducing a WOLF allele or a resistance-conferring part thereof into the genome of said spinach plant, or modifying an endogenous WOLF allele in the genome of said spinach plant. A WOLF allele encodes a CC-NBS- LRR protein that comprises in its amino acid sequence: the motif "MAEIGYSVC" at its N- terminus, and the motif "KWMCLR" for alpha-type WOLF proteins or "HVGCVVDR" for beta- type WOLF proteins. The invention relates to the WOLF alleles referred to in Table 3. The invention further provides a method for selecting a spinach plant comprising a novel WOLF gene that confers resistance to Peronospora farinosa f. sp. spinaciae in a spinach plant and a method for identifying a WOLF allele that confers resistance to one or more pathogenic races of Peronospora farinosa f. sp. spinaciae in a spinach plant and to primers for use in these methods.