Spinach Alpha-WOLF 15 Allele Broad-Spectrum Downy Mildew Resistance
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Solution Overview
Problem
Current spinach varieties are vulnerable to multiple races of Peronospora farinosa f. sp. spinaciae, leading to significant productivity losses due to the low durability of R-gene mediated resistance and the lack of characterized resistance genes, necessitating the development of new resistance alleles.
Innovation Solution
Identification and utilization of the alpha-WOLF 15 allele, a CC-NBS-LRR protein, which confers resistance to several downy mildew races by encoding a protein with specific motifs and sequence similarity, allowing for the development of resistant spinach plants through molecular biological tools and breeding methods.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If R-gene mediated resistance is used in spinach varieties, then resistance to downy mildew is achieved, but the durability of resistance is low and susceptibility to multiple races occurs
Solution Approach 1:
The patent identifies and utilizes the alpha-WOLF 15 allele that provides universal resistance across multiple Peronospora farinosa races (Pfs:1-Pfs:16). This single allele serves multiple functions by conferring broad-spectrum resistance against diverse downy mildew races, replacing the need for multiple race-specific R-genes and resolving the contradiction between durability and susceptibility to multiple races.
Solution Approach 2:
The patent employs molecular biological tools to identify and characterize the alpha-WOLF 15 allele through sequence analysis and genetic markers. By changing the parameter of resistance gene identification from phenotypic screening to genotypic characterization, the patent enables precise selection of alleles with durable broad-spectrum resistance properties.
2Reliability
If new resistance alleles are developed through breeding programs, then resistance to emerging isolates is achieved, but the process is time-consuming and complex
Solution Approach 1:
The patent performs preliminary characterization of the alpha-WOLF 15 allele including sequence determination, motif identification (CC-NBS-LRR domains), and resistance profiling against multiple races before deployment. This preliminary action creates a foundation for rapid breeding program implementation, as the allele's properties are already validated and characterized, reducing the time needed for subsequent breeding efforts.
Solution Approach 2:
The patent replaces traditional mechanical phenotypic screening methods with molecular biological tools for allele identification and characterization. By using DNA sequencing, bioinformatic analysis, and molecular markers instead of time-consuming field trials and visual assessment, the patent accelerates the development and deployment of resistance alleles.
3Measurement precision
If resistance genes are characterized at molecular level, then precise identification is achieved, but the current lack of characterization necessitates new research
Solution Approach 1:
The patent uses bioinformatic tools and databases as intermediaries to analyze and characterize the alpha-WOLF 15 allele. By employing computational methods to predict protein domains, motifs, and functional properties from sequence data, the patent achieves molecular-level characterization without requiring complex wet-lab infrastructure for every analysis step.
Solution Approach 2:
The patent creates detailed molecular copies and models of the alpha-WOLF 15 allele including its DNA sequence, predicted protein structure, and domain architecture. These digital copies enable precise identification and characterization while reducing the need for physical manipulation and complex experimental infrastructure.
Data Source
AI summary
The present invention relates to an allele designated alpha-WOLF 15 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 comprises in its amino acid sequence: a) the motif “MAEIGYSVC” (SEQ ID NO: 13) at its N-terminus; and b) the motif “KWMCLR” (SEQ ID NO: 14); 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: 10. The allele when homozygously present in a spinach plant confers complete resistance to Peronospora farinosa f. Sp. spinacea races pfs:1, pfs:2, pfs:3, pfs:4 and pfs:5, pfs:6, pfs:8, pfs:9, pfs:11, pfs:12, pfs:13, pfs:14, pfs:15, pfs:16 and isolates UA1014 and US1508, and confers intermediate resistance to pfs:10, and does not confer resistance to pfs:7.