基于液相芯片的大豆育成品种核心KASP标记体系构建与应用OA

To establish an efficient and streamlined molecular marker system dedicated to soybean,fulfill the demand for accurate variety identification and expand the repertoire of soybean molecular markers,this study took 115 released soybean varieties as experimental materials.Core SNP loci were screened via liquid chip data under the criteria of allelic frequency,chromosomal distribution and detection efficiency.A core KASP molecular marker system was then constructed,followed by performance evaluation of the system and the development of molecular identity cards for soybean varieties.The results showed that SNP genotyping was performed on a broad collection of soybean germplasm using the soybean 10 K liquid chip.A multi-stage hierarchical screening strategy was adopted based on polymorphic information content(PIC),minor allele frequency(MAF),sample call rate,even chromosomal distribution and clear genotyping performance.In the first-round screening,a total of 9 200 SNPs were retained from 30 323 SNPs under the thresholds of PIC>0.3,MAF>0.2 and sample call rate>0.95.After further selection,400 representative SNPs were retained,from which 238 KASP markers were developed via primer design.Subsequently,192 KASP markers with distinct genotyping profiles were obtained,and finally 60 core SNP markers were selected with stricter criteria of PIC>0.4 and uniform genome-wide distribution to establish a KASP marker system for soybean variety identification.The genetic distances calculated using these 60 markers showed an extremely significant linear correlation(r=0.622 1,P<0.01)with those derived from30 323 SNP loci of the soybean10 K liquid chip.Genotyping analysis of 115 soybean cultivars revealed that this system achieved a 100%variety discrimination efficiency,with pairwise genetic distances ranging from 0.100 to 0.799.Cluster analysis divided the 115 varieties into 5 groups,clearly reflecting their pedigree backgrounds and breeding histories.Additionally,a molecular identification system was established to realize digital labeling of varieties.The study demonstrates that this core KASP marker system effectively improves the accuracy and timeliness of soybean variety identification,authenticity testing,and DNA fingerprinting.It provides strong technical support for research on genetic diversity of soybean varieties,germplasm resource management,and protection of variety rights.