Application of Genome Reduction, Next Generation Sequencing, and KASPar Genotyping in Development, Characterization, and Linkage Mapping of Single Nucleotide Polymorphisms in the Grain Amaranths and Quinoa
The grain amaranths (Amaranthus sp.) and quinoa (Chenopodium quinoa Willd.) are important seed crops in South America. These crops have gained international attention in recent years for their nutritional quality and tolerance to abiotic stress. We report the identification and development of functional single nucleotide polymorphism (SNP) assays for both amaranth and quinoa. SNPs were identified using a genome reduction protocol and next generation sequencing. SNP assays are based on KASPar genotyping chemistry and were detected using the Fluidigm dynamic array platform. A diversity screen consisting of 41 amaranth accessions showed that the minor allele frequency (MAF) of the amaranth markers ranged from 0.05 to 0.5 with an average MAF of 0.27. A diversity screen of 113 quinoa accessions showed that the MAF of the quinoa markers ranged from 0.02 to 0.5 with an average MAF of 0.28. Linkage mapping in amaranth produced a linkage map consisting of 16 linkage groups, presumably corresponding to each of the 16 amaranth haploid chromosomes. This map spans 1288 cM with an average marker density of 3.1 cM per marker. Linkage mapping in quinoa resulted in a linkage map consisting of 29 linkage groups with 20 large linkage groups, spanning 1,404 cM with a marker density of 3.1 cM per SNP marker. The SNPs identified here represent important genomic tools needed for genetic dissection of agronomically important characteristics and advanced genetic analysis of agronomic traits in amaranth and quinoa. We also describe in detail the scalable and cost effective SNP genotyping method used in this research. This method is based on KBioscience's competitive allele specific PCR amplification of target sequences and endpoint fluorescence genotyping (KASPar) using a FRET capable plate reader or Fluidigm's dynamic array high throughput platform.
College and Department
Life Sciences; Plant and Wildlife Sciences
BYU ScholarsArchive Citation
Smith, Scott Matthew, "Application of Genome Reduction, Next Generation Sequencing, and KASPar Genotyping in Development, Characterization, and Linkage Mapping of Single Nucleotide Polymorphisms in the Grain Amaranths and Quinoa" (2013). Theses and Dissertations. 3548.
single nucleotide polymorphism, kaspar, amaranth, quinoa, genome reduction, next generation sequencing, snp genotyping