Spinach Downy Mildew Resistance via Alpha-WOLF 8 Allele

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Solution Overview

Problem

Current spinach varieties are vulnerable to multiple races of Peronospora farinosa f. sp. spinaciae, leading to significant losses in the spinach industry, as existing resistance genes are not durable and new races continually break through existing resistances.

Innovation Solution

Identification and utilization of the alpha-WOLF 8 allele, a CC-NBS-LRR protein-encoding gene that confers resistance to several downy mildew races, including Pfs:16, by amplifying and sequencing specific primers for the LRR domain, providing a molecular tool for breeding resistant spinach plants.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If existing resistance genes are used in spinach varieties, then resistance to current downy mildew races is achieved, but resistance durability is poor as new races continually break through existing resistances

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

Solution Approach 1:

The WOLF gene serves as a universal resistance gene that provides protection against multiple downy mildew races (Pfs:1, Pfs:2, Pfs:6, Pfs:8, Pfs:15, and intermediate resistance to Pfs:5, Pfs:10, and Pfs:16) through a single genetic locus, replacing the need for multiple race-specific resistance genes and enabling broad-spectrum durability

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

Solution Approach 2:

The invention utilizes allelic variation at the WOLF locus, where different alleles confer resistance to different races. By identifying and deploying specific alleles like WOLF 8, the resistance profile can be tailored to match emerging race patterns, dynamically adapting the resistance parameters to changing pathogen pressures

Inventive Principle:
Principle #35Parameter changes

2Adaptability or versatility

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

Engineering Contradiction:
Improveresistance spectrumVSAvoidgenetic complexity
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

A single WOLF gene allele provides resistance to multiple downy mildew races, eliminating the need to stack multiple resistance genes to achieve broad-spectrum coverage. This simplifies the genetic architecture from multiple genes to a single multi-functional gene locus

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

Solution Approach 2:

The invention merges the functions of multiple race-specific resistance genes into a single WOLF gene locus, where one gene performs what previously required multiple genes, thereby reducing genetic complexity while maintaining or expanding the resistance spectrum

Inventive Principle:
Principle #5Merging (Combining)

Data Source

PatentUS10674688B2<i>Peronospora </i>resistance in spinacia oleracea
Publication Date: 2020.06.09 RIJK ZWAAN ZAADTEELT & ZAADHANDEL BV

AI summary

The present invention relates to an allele designated alpha-WOLF 8 which confers resistance to at least one Peronospora farinosa f. sp. spinaciae race, wherein the protein encoded by said allele is a CC-NB S-LRR protein that comprises in its amino acid sequence: a) the motif “MAEIGYSVC” (SEQ ID NO: 15) at its N-terminus; and b) the motif “KWMCLR” (SEQ ID NO: 16); and wherein the LRR domain of the protein has in order of increased preference at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence similarity to SEQ ID NO: 12. When the allele is homozygously present in a spinach plant confers complete resistance to Peronospora farinosa f. sp. spinaciae races Pfs:1, Pfs:2, Pfs:6, Pfs:8 and Pfs:15, and confers intermediate resistance to Pfs:5, Pfs:10 and Pfs:16, and does not confer resistance to Pfs:3, Pfs:4, Pfs:7, Pfs:9, Pfs:11, Pfs:12, Pfs:13 and Pfs:14.