ABCB9 Gene Variants for CMV-Resistant Cucurbit Breeding
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Solution Overview
Problem
There is no known gene responsible for the genetic basis of Cucumber Mosaic Virus (CMV) resistance in plants, particularly in Cucurbitaceae species like cucumber and melon, despite some sources of resistance existing.
Innovation Solution
Identification of modified ABCB9 and EF1-α genes with specific nucleotide substitutions that confer CMV resistance when homozygously present in plants, particularly cucumber plants, with the ABCB9 gene having a SNP at position 1282 and EF1-α gene having a SNP at position 1115, leading to amino acid changes that enhance resistance.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If no gene modification is applied, then the plant maintains its natural genetic composition, but the plant shows susceptibility to CMV infection
Solution Approach 1:
The invention applies parameter changes by modifying specific nucleotide positions (position 1282 in ABCB9 and position 1115 in EF1-α) to create SNP variants that confer CMV resistance. This targeted nucleotide substitution changes the genetic parameters of the plant to achieve the desired resistance trait while maintaining overall genetic compatibility.
Solution Approach 2:
The invention uses marker-assisted selection to identify and copy plants with the desired SNP modifications. Specific markers detect the presence of modified ABCB9 and EF1-α genes, allowing breeders to copy and propagate the resistant genetic variants through selective breeding without directly observing phenotypic resistance.
2Reliability
If modified ABCB9 and EF1-α genes are introduced, then CMV resistance is enhanced, but the difficulty of detecting and measuring the resistance increases
Solution Approach 1:
The invention creates and uses specific genetic markers that copy the detection capability from the complex modified genes to simpler detectable signals. These markers serve as proxies for the actual resistance-conferring SNPs, making detection straightforward through standard molecular biology techniques rather than requiring complex phenotypic assays.
Solution Approach 2:
The invention introduces marker genes as intermediaries between the actual resistance-conferring SNPs and the detection process. These markers are genetically linked to the modified ABCB9 and EF1-α genes and serve as measurable indicators that mediate the detection of resistance without requiring direct measurement of the resistance mechanism itself.
3Reliability
If homozygous presence of both modified genes is required for maximum resistance, then resistance reliability is maximized, but the ease of manufacture and breeding becomes more difficult
Solution Approach 1:
The invention implements feedback through marker-assisted selection at multiple breeding stages. Breeders can detect the presence of modified genes in heterozygous and homozygous states using specific markers, providing feedback that guides selection decisions. This allows systematic progression from heterozygous to homozygous plants across generations, making the breeding process more predictable and efficient.
Solution Approach 2:
The invention applies preliminary action by using markers to identify and select plants carrying the modified genes before phenotypic resistance can be fully assessed. This allows breeders to pre-select plants with the desired genetic modifications in early generations, reducing the time and effort required to develop homozygous resistant lines compared to traditional phenotypic selection methods.
Data Source
AI summary
The present invention relates to a modified ABCB9 gene encoding a protein conferring resistance to CMV in a plant of the Cucurbitaceae, such as a cucumber plant, in which the protein is expressed, where the gene has a) a nucleotide sequence which encodes a protein with SEQ ID NO: 4; b) a nucleotide sequence with SEQ ID NO: 3; c) a nucleotide sequence encoding a protein derived by substitution, deletion and/or addition of one or more amino acids of the protein with SEQ ID NO: 4; d) a nucleotide sequence that encodes a protein with an amino acid sequence, which is at least 85% identical to SEQ ID NO: 4; e) a nucleotide sequence which is at least 85% identical to SEQ ID NO: 3; or f) a nucleotide sequence according to c) or d) wherein the protein has a methionine (M) on position 428 of SEQ ID NO: 4, or on a position corresponding thereto.