A comparison of simple sequence repeat and single nucleotide polymorphism marker technologies for the genotypic analysis of maize (Zea mays L.)

摘要

We report on the comparative utilities of simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers for characterizing maize germplasm in terms of their informativeness, levels of missing data, repeatability and the ability to detect expected alleles in hybrids and DNA pools. Two different SNP chemistries were compared; single-base extension detected by Sequenom MassARRAY, and invasive cleavage detected by Invader chemistry with PCR. A total of 58 maize inbreds and four hybrids were genotyped with 80 SSR markers, 69 Invader SNP markers and 118 MassARRAY SNP markers, with 64 SNP loci being common to the two SNP marker chemistries. Average expected heterozygosity values were 0.62 for SSRs, 0.43 for SNPs (pre-selected for their high level of polymorphism) and 0.63 for the underlying sequence haplotypes. All individual SNP markers within the same set of sequences had an average expected heterozygosity value of 0.26. SNP marker data had more than a fourfold lower level of missing data (2.1-3.1%) compared with SSRs (13.8%). Data repeatability was higher for SNPs (98.1% for MassARRAY SNPs and 99.3% for Invader) than for SSRs (91.7%). Parental alleles were observed in hybrid genotypes in 97.0% of the cases for MassARRAY SNPs, 95.5% for Invader SNPs and 81.9% for SSRs. In pooled samples with mixtures of alleles, SSRs, MassARRAY SNPs and Invader SNPs were equally capable of detecting alleles at mid to high frequencies. However, at low frequencies, alleles were least likely to be detected using Invader SNP markers, and this technology had the highest level of missing data. Collectively, these results showed that SNP technologies can provide increased marker data quality and quantity compared with SSRs. The relative loss in polymorphism compared with SSRs can be compensated by increasing SNP numbers and by using SNP haplotypes. Determining the most appropriate SNP chemistry will be dependent upon matching the technical features of the method within the context of application, particularly in consideration of whether genotypic samples will be pooled or assayed individually.