Jones E S, Sullivan H, Bhattramakki D, Smith J S C
Pioneer Hi-Bred International Inc. (DuPont Agriculture and Nutrition), 7300 NW 62nd Avenue, Johnston, IA 51031-1004, USA.
Theor Appl Genet. 2007 Aug;115(3):361-71. doi: 10.1007/s00122-007-0570-9. Epub 2007 May 22.
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.
我们报告了简单序列重复(SSR)和单核苷酸多态性(SNP)标记在玉米种质特征鉴定方面的比较效用,包括信息性、缺失数据水平、重复性以及在杂交种和DNA混合池中检测预期等位基因的能力。比较了两种不同的SNP化学方法;通过Sequenom MassARRAY检测的单碱基延伸法,以及通过带有PCR的Invader化学法检测的侵入性切割法。对58个玉米自交系和4个杂交种进行了基因分型,使用了80个SSR标记、69个Invader SNP标记和118个MassARRAY SNP标记,两种SNP标记化学方法共有64个SNP位点。SSR的平均期望杂合度值为0.62,SNP(因其高度多态性而预先选择)为0.43,潜在序列单倍型为0.63。同一组序列内的所有单个SNP标记的平均期望杂合度值为0.26。与SSR(13.8%)相比,SNP标记数据的缺失数据水平低四倍多(2.1 - 3.1%)。SNP的数据重复性高于SSR(MassARRAY SNP为98.1%,Invader为99.3%,SSR为91.7%)。在杂交种基因型中,97.0%的情况可观察到MassARRAY SNP的亲本等位基因,95.5%的情况可观察到Invader SNP的亲本等位基因,81.9%的情况可观察到SSR的亲本等位基因。在具有等位基因混合物的混合样本中,SSR、MassARRAY SNP和Invader SNP在检测中高频等位基因方面同样有效。然而,在低频时,使用Invader SNP标记检测等位基因的可能性最小,且该技术的缺失数据水平最高。总体而言,这些结果表明,与SSR相比,SNP技术可提供更高质量和数量的标记数据。与SSR相比多态性的相对损失可通过增加SNP数量和使用SNP单倍型来弥补。确定最合适的SNP化学方法将取决于在应用背景下匹配方法的技术特征,特别是考虑基因型样本是混合还是单独检测。