Soybean KASP Marker Genotyping for Oligosaccharide Content
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Solution Overview
Problem
Current methods for screening soybean oligosaccharide content are time-consuming, costly, and laborious, failing to meet demands for efficient and stable identification, especially due to their sensitivity to environmental factors.
Innovation Solution
Development of a soybean oligosaccharide-related KASP marker, including specific SNP sites and primer pairs (S18_51868868 T/G and S10_38081012 T/C), which allow for accurate genotyping of raffinose and stachyose content through PCR amplification and fluorescence signal analysis, enabling efficient breeding of high oligosaccharide soybeans.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional screening methods are used to determine oligosaccharide content of each soybean germplasm, then identification can be performed, but the identification period is long, cost is high, and the process is time-consuming and laborious
Solution Approach 1:
The patent replaces conventional mechanical/chemical screening methods with molecular marker-based genotyping. By using KASP markers associated with oligosaccharide content, the system substitutes direct phenotypic measurement with genotypic analysis, achieving rapid identification without time-consuming cultivation and chemical analysis.
Solution Approach 2:
The patent uses molecular markers as copies or proxies for the actual oligosaccharide traits. Instead of measuring the oligosaccharide content directly, the system detects genetic variants (SNPs) that correlate with high oligosaccharide content, creating an indirect but efficient identification method through genetic copying.
2Measurement precision
If conventional screening methods are used to determine oligosaccharide content, then identification can be performed, but the cost is high and the process is laborious
Solution Approach 1:
The patent replaces complex conventional screening procedures with a streamlined molecular genotyping system. The KASP marker assay substitutes multiple labor-intensive steps (planting, harvesting, chemical extraction, HPLC analysis) with a single PCR-based genotyping reaction, dramatically simplifying the process.
Solution Approach 2:
The patent extracts the essential identification function from the complex conventional screening process. By isolating and utilizing specific molecular markers associated with oligosaccharide content, the system removes unnecessary steps and retains only the critical identification function, making the process simpler and more efficient.
3Reliability
If conventional screening methods are used, then oligosaccharide content can be determined, but the method cannot meet the demands of stable and efficient screening due to environmental factor sensitivity
Solution Approach 1:
The patent inverts the conventional approach by moving from phenotypic screening (measuring oligosaccharide content in mature plants) to genotypic screening (detecting genetic markers). This inversion allows identification before environmental factors can influence the trait expression, ensuring stable and reliable results independent of environmental conditions.
Solution Approach 2:
The patent performs preliminary identification at the genetic level before environmental factors can affect oligosaccharide accumulation. By detecting molecular markers associated with high oligosaccharide content early in the breeding process, the system establishes reliable identification that is not influenced by subsequent environmental variations during plant growth.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The KASP markers achieve high accuracy rates (91.3% for raffinose and 93.5% for stachyose) in detecting oligosaccharide content, facilitating faster and more efficient soybean breeding by distinguishing genotypes with higher or lower oligosaccharide levels, thus improving the breeding process.
Implementation Method 1
conducting PCR amplification using a genomic DNA of a soybean to be tested as a template with the primer pair
Implementation Method 2
genotyping according to a fluorescence signal of a PCR amplification product
Data Source
AI summary
Provided is a soybean oligosaccharide-related kompetitive allele-specific PCR (KASP) marker and use thereof, belonging to the technical field of molecular breeding. The KASP marker includes one or two of S18_51868868 T/G and S10_38081012 T/C, where the S18_51868868 T/G is a base T or a base G at a 51868868bp position on chromosome 18 of a soybean genome; and the S10_38081012 T/C is a base T or a base C at a 38081012bp position of chromosome 10 of the soybean genome. In the present disclosure, the KASP marker can accurately genotype the traits of raffinose and stachyose contents. Through genotyping, it is found that a soybean germplasm with a genotype GG has a higher raffinose content than that of a soybean germplasm with a genotype TT, and a soybean germplasm with a genotype TT has a higher stachyose content than that of a soybean germplasm with a genotype CC.