Temperate Tree Fruit and Vegetable Research Unit, U.S. Department of Agriculture Agricultural Research Service, Prosser, WA 99350.
Grain Legume Genetics and Physiology Research Unit, U.S. Department of Agriculture Agricultural Research Service, Prosser, WA 99350.
Plant Dis. 2020 Jun;104(6):1824-1830. doi: 10.1094/PDIS-09-19-1920-RE. Epub 2020 Apr 9.
As pesticides have become heavily relied on for management of insect pests vectoring economically important pathogens of vegetable crops, development of pathogen-resistant germplasm remains a promising alternative to reduce or eliminate costly and timely chemical inputs. Molecular markers can be used to rapidly identify resistant genotypes to aid breeders in advancing germplasm. This study developed two kompetitive allele-specific PCR (KASP) genotyping markers for rapid screening of genotypes for resistance to pathotype P1 (PSbMV-P1), the most economically devastating strain worldwide. The KASP markers differentiate two PSbMV-P1-resistant allelic variants from susceptible variants. A single nucleotide polymorphism (Resistant 1) and a 3-basepair deletion (Resistant 2) present in either of the two resistant alleles were used for marker design. Forty-four lines previously characterized for resistance to PSbMV were inoculated with PSbMV-P1 in a greenhouse, observed for visual symptoms, assayed for virus susceptibility by enzyme-linked immunosorbent assay (ELISA), and genotyped by KASP marker analysis. The KASP markers were 100% accurate in characterizing PSbMV-P1-susceptible and PSbMV-P1-resistant genotypes when correlated with the ELISA results. The Resistant 1 marker also correlated with resistance to PSbMV pathotypes P2 and P4 completely, making this marker a new advanced tool for breeding programs.
由于杀虫剂在防治传播蔬菜作物重要经济病原菌的害虫方面已被大量依赖,因此培育具有病原体抗性的种质资源仍然是减少或消除昂贵且及时的化学投入的一种很有前景的替代方法。分子标记可用于快速鉴定抗性基因型,以帮助培育者推进种质资源的发展。本研究开发了两种竞争性等位基因特异性 PCR(KASP)基因分型标记,用于快速筛选对 路径型 P1(PSbMV-P1)具有抗性的基因型,PSbMV-P1 是全球经济破坏性最强的菌株。KASP 标记可区分两种对 PSbMV-P1 具有抗性的等位基因变体和易感变体。用于标记设计的是两个抗性等位基因中存在的单个核苷酸多态性(抗性 1)和 3 个碱基对缺失(抗性 2)。先前已对 44 个 PSbMV 抗体制品进行了鉴定,在温室中用 PSbMV-P1 接种这些抗体制品,观察其视觉症状,通过酶联免疫吸附试验(ELISA)测定其对病毒的敏感性,并通过 KASP 标记分析进行基因型鉴定。当与 ELISA 结果相关联时,KASP 标记在鉴定 PSbMV-P1 敏感和 PSbMV-P1 抗性基因型时的准确率达到 100%。抗性 1 标记也与 PSbMV 路径型 P2 和 P4 的抗性完全相关,使该标记成为 育种计划的新先进工具。