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通过测序进行基因分型实现菜豆中BY-2马铃薯Y病毒抗性等位基因的定位与标记开发

Genotyping-by-Sequencing Enabled Mapping and Marker Development for the By-2 Potyvirus Resistance Allele in Common Bean.

作者信息

Hart John P, Griffiths Phillip D

机构信息

USDA-ARS, Tropical Agriculture Res. Stn. (TARS), 2200 P.A. Campos Ave., Suite 201, Mayagüez, PR 00680-5470.

School of Integrative Plant Science, Plant Breeding and Genetics Section, Cornell Univ., 314 Hedrick Hall, Geneva, NY, 14456.

出版信息

Plant Genome. 2015 Mar;8(1):eplantgenome2014.09.0058. doi: 10.3835/plantgenome2014.09.0058.

Abstract

Since its emergence in 2001, an aphid-transmitted virus disease complex has caused substantial economic losses to snap bean (Phaseolus vulgaris L.) production and processing in the Great Lakes Region of the United States. The general ineffectiveness of chemical control measures for nonpersistently transmitted viruses established an urgent need for the development and deployment of cultivars with resistance to the component viruses. Our objectives were to further characterize the inheritance of resistance to Bean yellow mosaic virus (BYMV), which is conditioned by the By-2 allele, to adapt genotyping-by-sequencing (GBS) to common bean to discover and genotype genome-wide single nucleotide polymorphisms (SNPs) in a set of recombinant inbred lines (RILs) derived from an introgression program, and to enable and validate marker-assisted selection for By-2. We optimized ApeKI for GBS in common bean and retained 7530 high-quality SNPs that segregated in our introgression RILs. A case-control genome-wide association study (GWAS) was used to discover 44 GBS SNPs that were strongly associated with the resistance phenotype and which delimited a 974 kb physical interval on the distal portion of chromosome 2. Seven of these SNPs were converted to single-marker Kompetitive Allele-Specific Polymerase chain reaction (KASP) assays and were demonstrated to be tightly linked to BYMV resistance in an F population of 185 individuals. This research enables marker-assisted selection of By-2, provides enhanced resolution for fine mapping, and demonstrates the potential of GBS as a highly efficient, high-throughput genotyping platform for common bean breeding and genetics.

摘要

自2001年出现以来,一种由蚜虫传播的病毒病复合体已给美国大湖地区的菜豆(Phaseolus vulgaris L.)生产和加工造成了巨大经济损失。化学防治措施对非持久性传播病毒普遍无效,这迫切需要开发和推广对组成病毒具有抗性的品种。我们的目标是进一步表征由By-2等位基因决定的对菜豆黄花叶病毒(BYMV)抗性的遗传特性,使测序基因分型(GBS)适用于普通菜豆,以便在一组源自渐渗计划的重组自交系(RIL)中发现全基因组单核苷酸多态性(SNP)并进行基因分型,并实现和验证对By-2的标记辅助选择。我们优化了普通菜豆GBS中的ApeKI,并保留了7530个在我们的渐渗RIL中分离的高质量SNP。采用病例对照全基因组关联研究(GWAS)来发现44个与抗性表型强烈相关的GBS SNP,这些SNP在2号染色体远端界定了一个974 kb的物理区间。其中7个SNP被转化为单标记竞争性等位基因特异性聚合酶链反应(KASP)分析,并在一个由185个个体组成的F群体中被证明与BYMV抗性紧密连锁。这项研究实现了对By-2的标记辅助选择,提高了精细定位的分辨率,并证明了GBS作为普通菜豆育种和遗传学中一种高效、高通量基因分型平台的潜力。

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