Genetic Marker Generation via Random Mutagenesis for Plant Breeding

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Solution Overview

Problem

Marker-assisted selection in plant breeding is hindered by the challenge of identifying relevant markers, especially for traits encoded in regions of low polymorphism or complex quantitative trait loci, where markers may not be unique or closely linked, leading to issues like linkage drag.

Innovation Solution

The method involves de novo generation and introduction of genetic markers through random mutagenesis, allowing for the identification and selection of markers linked to genomic regions of interest, even in areas of low polymorphism, without prior knowledge of the trait's location, thereby fine-mapping the locus and minimizing linkage drag.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If phenotypic screening and selection is used to identify plants with desired traits, then accurate trait evaluation can be achieved, but the process becomes cumbersome and time-consuming as plants must be grown to maturity

Engineering Contradiction:
Improvetrait evaluation accuracyVSAvoidbreeding cycle duration
Core Design Contradiction:
Measurement precisionVSLoss of time

Solution Approach 1:

The patent applies preliminary action by introducing mutations and creating genetic markers in advance, before phenotypic evaluation is needed. These pre-established markers can then be used for rapid selection without waiting for plants to mature, thus reducing breeding cycle time while maintaining selection accuracy

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent uses genetic markers as an intermediary between the trait of interest and the selection process. Instead of directly evaluating the trait phenotype, markers serve as proxies that indicate the presence of desired traits, enabling indirect but accurate selection without time-consuming phenotypic assessment

Inventive Principle:
Principle #24Intermediary (Mediator)

2Productivity

If markers are used for early stage selection, then breeding efficiency is improved, but relevant markers are difficult to identify especially in regions of low polymorphism or complex QTL

Engineering Contradiction:
Improvebreeding efficiencyVSAvoidmarker identification difficulty
Core Design Contradiction:
ProductivityVSDifficulty of detecting and measuring

Solution Approach 1:

The patent segments the challenge of marker identification by focusing on creating markers through random mutagenesis in specific genomic regions rather than searching for existing natural polymorphisms. This approach divides the problem into manageable steps: induce mutations, screen for markers, and validate linkage

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent applies preliminary action by proactively introducing mutations to create markers before the breeding selection process begins. This eliminates the need to search for pre-existing markers in low-polymorphism regions, as markers are generated de novo through mutagenesis

Inventive Principle:
Principle #10Preliminary action

3Measurement precision

If markers are used to select against linkage drag, then selection precision is improved, but markers must be in close proximity to the trait which limits available options

Engineering Contradiction:
Improveselection precisionVSAvoidmarker availability
Core Design Contradiction:
Measurement precisionVSAdaptability or versatility

Solution Approach 1:

The patent applies preliminary action by introducing multiple random mutations across the genome before selection. This creates a pool of potential markers at various distances from target traits, providing versatility in marker selection while ensuring some markers will be in close proximity for precise selection against linkage drag

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent uses partial or excessive action by introducing more mutations than strictly necessary. This excess mutagenesis ensures that sufficient markers are created in close proximity to all relevant traits, providing multiple options for precise selection and overcoming the limitation of marker availability

Inventive Principle:
Principle #16Partial or excessive action

Data Source

PatentEP3918911A1Marker generation by random mutagenesis in plants
Publication Date: 2021.12.08 KWS SAAT SE & CO KGAA
  • EP3918911A1 patent drawingFigure 1
  • EP3918911A1 patent drawingFigure 2
  • EP3918911A1 patent drawingFigure 3

AI summary

The present invention relates to a method for introducing mutations in the nucleotide sequence of a 5' or 3' flanking region of a genomic region of interest in plants by chemical treatment or radiation. These introduced mutations are used to develop genetic markers, which are linked to said genomic region of interest and allow for precise selection thereof. The present invention further relates to a method for generating such genetic markers, a method for detecting a genomic region of interest with said genetic markers as well as to a method for identifying plants comprising such genetic markers.