Muchero Wellington, Diop Ndeye N, Bhat Prasanna R, Fenton Raymond D, Wanamaker Steve, Pottorff Marti, Hearne Sarah, Cisse Ndiaga, Fatokun Christian, Ehlers Jeffrey D, Roberts Philip A, Close Timothy J
Departments of Nematology and Botany and Plant Sciences, University of California, Riverside, CA 92521, USA.
Proc Natl Acad Sci U S A. 2009 Oct 27;106(43):18159-64. doi: 10.1073/pnas.0905886106. Epub 2009 Oct 13.
Consensus genetic linkage maps provide a genomic framework for quantitative trait loci identification, map-based cloning, assessment of genetic diversity, association mapping, and applied breeding in marker-assisted selection schemes. Among "orphan crops" with limited genomic resources such as cowpea [Vigna unguiculata (L.) Walp.] (2n = 2x = 22), the use of transcript-derived SNPs in genetic maps provides opportunities for automated genotyping and estimation of genome structure based on synteny analysis. Here, we report the development and validation of a high-throughput EST-derived SNP assay for cowpea, its application in consensus map building, and determination of synteny to reference genomes. SNP mining from 183,118 ESTs sequenced from 17 cDNA libraries yielded approximately 10,000 high-confidence SNPs from which an Illumina 1,536-SNP GoldenGate genotyping array was developed and applied to 741 recombinant inbred lines from six mapping populations. Approximately 90% of the SNPs were technically successful, providing 1,375 dependable markers. Of these, 928 were incorporated into a consensus genetic map spanning 680 cM with 11 linkage groups and an average marker distance of 0.73 cM. Comparison of this cowpea genetic map to reference legumes, soybean (Glycine max) and Medicago truncatula, revealed extensive macrosynteny encompassing 85 and 82%, respectively, of the cowpea map. Regions of soybean genome duplication were evident relative to the simpler diploid cowpea. Comparison with Arabidopsis revealed extensive genomic rearrangement with some conserved microsynteny. These results support evolutionary closeness between cowpea and soybean and identify regions for synteny-based functional genomics studies in legumes.
一致性遗传连锁图谱为数量性状基因座鉴定、基于图谱的克隆、遗传多样性评估、关联作图以及标记辅助选择育种方案中的应用育种提供了基因组框架。在诸如豇豆[Vigna unguiculata (L.) Walp.](2n = 2x = 22)等基因组资源有限的“孤儿作物”中,利用转录本衍生的单核苷酸多态性(SNP)构建遗传图谱为基于共线性分析的自动化基因分型和基因组结构估计提供了机会。在此,我们报告了一种针对豇豆的高通量EST衍生SNP检测方法的开发与验证、其在构建一致性图谱中的应用以及与参考基因组的共线性测定。从17个cDNA文库测序得到的183,118个EST中挖掘SNP,得到了约10,000个高可信度SNP,据此开发了一种Illumina 1,536-SNP金标准基因分型芯片,并应用于来自六个作图群体的741个重组自交系。约90%的SNP在技术上成功,提供了1,375个可靠标记。其中,928个被整合到一个包含11个连锁群、跨度为680 cM、平均标记间距为0.73 cM的一致性遗传图谱中。将该豇豆遗传图谱与参考豆科植物大豆(Glycine max)和蒺藜苜蓿(Medicago truncatula)进行比较,发现分别有85%和82%的豇豆图谱区域存在广泛的宏观共线性。相对于较为简单的二倍体豇豆,大豆基因组重复区域明显。与拟南芥的比较揭示了广泛的基因组重排以及一些保守的微观共线性。这些结果支持了豇豆与大豆之间的进化亲缘关系,并确定了豆科植物中基于共线性的功能基因组学研究区域。