Individual Microspore Genotyping in Sorghum (S. bicolor) Cultivars Using a KASP Panel of Single Nucleotide Polymorphism Markers

Dilovan K. Yahya, Matthew W. Blair, Aron M. Felts, Ahmad N. Aziz

Abstract


Sweet sorghum (Sorghum bicolor L. Moench) cultivars show promise as a biofuel source, necessitating innovative tools for genetic enhancements. This study employed whole genome amplification (WGA) to amplify genomic DNA from individually isolated haploid microspores. These microspores were genotyped using a panel of novel, locus-specific, single nucleotide polymorphism (SNP) markers. Five sorghum cultivars were grown to the flowering stage, and 30 early developing microspores were individually isolated from each variety. Kompetitive Allele Specific PCR (KASP) based markers' analysis was performed on 20 SNP loci from all varieties using microspore and leaf DNAs. The KASP success rate was 80% for microspore DNAs and 91.3% for leaf samples. Eight SNPs detected 16-42% mono-allelic markers in microspores while the other 12 detected mono-allelism at 10% or less. Microspores that displayed similar profiles were grouped together for each variety. Notably, 13 SNPs were selected for such gametic groups, demonstrating the utility of WGA products from individual gametophytes. This study marks the first application of molecular markers using individually isolated microspores of sorghum varieties.


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