The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Development and Validation of a Perfect KASP Marker for Fusarium Head Blight Resistance Gene Fhb1 in Wheat

  • Singh, Lovepreet (Department of Plant Science and Landscape Architecture, University of Maryland) ;
  • Anderson, James A (Department of Agronomy and Plant Genetics, University of Minnesota) ;
  • Chen, Jianli (Department of Plant, Soil, and Entomological Sciences, University of Idaho) ;
  • Gill, Bikram S (Wheat Genetics Resource Center, Department of Plant Pathology, Kansas State University) ;
  • Tiwari, Vijay K (Department of Plant Science and Landscape Architecture, University of Maryland) ;
  • Rawat, Nidhi (Department of Plant Science and Landscape Architecture, University of Maryland)
  • Received : 2019.01.27
  • Accepted : 2019.03.31
  • Published : 2019.06.01
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Abstract

Fusarium head blight (FHB) is a devastating wheat disease with a significant economic impact. Fhb1 is the most important large effect and stable QTL for FHB resistance. A pore-forming toxin-like (PFT) gene was recently identified as an underlying gene for Fhb1 resistance. In this study, we developed and validated a PFT-based Kompetitive allele specific PCR (KASP) marker for Fhb1. The KASP marker, PFT_KASP, was used to screen 298 diverse wheat breeding lines and cultivars. The KASP clustering results were compared with gelbased gene specific markers and the widely used linked STS marker, UMN10. Eight disagreements were found between PFT_KASP and UMN10 assays among the tested lines. Based on the genotyping and sequencing of genes in the Fhb1 region, these genotypes were found to be common with a previously characterized susceptible haplotype. Therefore, our results indicate that PFT_KASP is a perfect diagnostic marker for Fhb1 and would be a valuable tool for introgression and pyramiding of FHB resistance in wheat cultivars.

Keywords

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Fig. 1. Multiple sequence alignment of wheat accessions representing R and S3 haplotypes for PFT. WZHHS, Shanasui, Wannin2, Sumai-3, Huoshawmai, Shuilizhan, and 701 Chokwang contain R haplotype, whereas, Jingzhou1, Nanda2419, and Emai6 possess S3 haplotype. Asterisks designate the consensus sequence among all the genotypes. The position of allele specific and common primer is highlighted as bold underlined. The SNP targeted for developing PFT_KASP is highlighted as bold italics.

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Fig. 3. Order and location of genes and markers on Fhb1 genomic assembly (Rawat et al., 2016) used to resolve the haplotypes of the eight breeding lines/cultivars with disagreement between UMN10 and PFT.

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Fig. 2. Genotyping results of PFT_KASP marker on a panel of different genotypes and controls. Genotypes clustered as blue squares in the upper left corner are homozygous resistant (R) haplotype. Genotypes clustered in the lower right corner as orange circles are homozygous susceptible (S3) haplotype. Genotypes which do not possess either allele are clustered with NTCs (No template controls) as black diamonds in the lower left corner.

Table 1. List of primer sequences used in this study

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Table 2. Genotyping results of PFT_KASP, UMN10, SGNH_GSP, and PFT_GSP markers on the set of University of Idaho wheat breeding lines used for validation of PFT_KASP marker

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Table 2. Continued

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Table 3. Haplotypes of genes spanning Fhb1 region in the eight lines showing disagreements between PFT_KASP and UMN10 genotyping

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