Development and Application of a High-Efficiency 1.8K SNP Liquid Chip for Tobacco Breeding Based on Multiplex PCR Sequencing
Based on the publicly available tobacco K326 genome and data from the 430K SNP solid-phase chip for tobacco,this study developed a tobacco 1.8K breeding liquid-phase chip(YT1.8K.1)targeting 20 polymorphic markers on each chromosome between seven pairs of germplasm combinations,utilizing multiplex PCR amplification and pinpoint sequencing-based genotyping technology(mGPS,Genotyping by Pinpoint Sequencing of multiplex PCR products).This chip was used for genotyping in 21 hybrid combinations of the seven germplasms mentioned,with an average of 650 differential loci between each combination,effectively supporting the selective breeding for high genetic background recovery rate in each combination.The chip was further used for genotyping and cluster analysis of 23 tobacco varieties,where the clustering classification results aligned well with the their pedigree information.Additionally,The chip also successfully identified five individual plants with background recovery rates above 94.96%out of 367 BC2F1 populations,surpassing the theoretical mean of 87.5%,indicating that this breeding chip can be applied to cluster analysis of tobacco germplasm resources and genetic background selection for targeted improvement breeding.
万方数据
维普
