Bertioli David J, Ozias-Akins Peggy, Chu Ye, Dantas Karinne M, Santos Silvio P, Gouvea Ediene, Guimarães Patricia M, Leal-Bertioli Soraya C M, Knapp Steven J, Moretzsohn Marcio C
Departamento de Genética/IB, Universidade de Brasília, Brasília, DF, 70910-900, Brazil.
G3 (Bethesda). 2014 Jan 10;4(1):89-96. doi: 10.1534/g3.113.007617.
Single nucleotide polymorphic markers (SNPs) are attractive for use in genetic mapping and marker-assisted breeding because they can be scored in parallel assays at favorable costs. However, scoring SNP markers in polyploid plants like the peanut is problematic because of interfering signal generated from the DNA bases that are homeologous to those being assayed. The present study used a previously constructed 1536 GoldenGate SNP assay developed using SNPs identified between two A. duranensis accessions. In this study, the performance of this assay was tested on two RIL mapping populations, one diploid (A. duranensis × A. stenosperma) and one tetraploid [A. hypogaea cv. Runner IAC 886 × synthetic tetraploid (A. ipaënsis × A. duranensis)(4×)]. The scoring was performed using the software GenomeStudio version 2011.1. For the diploid, polymorphic markers provided excellent genotyping scores with default software parameters. In the tetraploid, as expected, most of the polymorphic markers provided signal intensity plots that were distorted compared to diploid patterns and that were incorrectly scored using default parameters. However, these scorings were easily corrected using the GenomeStudio software. The degree of distortion was highly variable. Of the polymorphic markers, approximately 10% showed no distortion at all behaving as expected for single-dose markers, and another 30% showed low distortion and could be considered high-quality. The genotyped markers were incorporated into diploid and tetraploid genetic maps of Arachis and, in the latter case, were located almost entirely on A genome linkage groups.
单核苷酸多态性标记(SNPs)因其能以合理成本在平行分析中进行评分,故而在遗传图谱构建和标记辅助育种中颇具吸引力。然而,在花生这类多倍体植物中对SNP标记进行评分存在问题,因为与被检测的DNA碱基同源的碱基会产生干扰信号。本研究使用了先前构建的1536个金门SNP分析方法,该方法是利用两个A. duranensis种质间鉴定出的SNP开发的。在本研究中,对两个重组自交系作图群体测试了该分析方法的性能,一个是二倍体群体(A. duranensis×A. stenosperma),另一个是四倍体群体[A. hypogaea cv. Runner IAC 886×人工合成四倍体(A. ipaënsis×A. duranensis)(4x)]。使用GenomeStudio 2011.1版本软件进行评分。对于二倍体群体,多态性标记在默认软件参数下提供了出色的基因分型评分。在四倍体群体中,正如预期的那样,与二倍体模式相比,大多数多态性标记提供的信号强度图出现了扭曲,并且使用默认参数进行评分时出现错误。然而,使用GenomeStudio软件可以轻松校正这些评分。扭曲程度变化很大。在多态性标记中,约10%完全没有扭曲,表现如同单剂量标记所预期的那样,另外30%显示出低扭曲,可以被认为是高质量的。将基因分型标记整合到花生的二倍体和四倍体遗传图谱中,在四倍体图谱的情况下,这些标记几乎完全位于A基因组连锁群上。