Development and application of a cost-effective multiplex Kompetitive Allele-Specific polymerase chain reaction assay for pyramiding resistant genes of fusarium head blight and powdery mildew in wheat
Abstract Background Wheat (Triticum aestivum L.) cultivation suffers from significant yield loss owing to diseases such as Fusarium head blight (FHB) and powdery mildew (PM). Utilization of host resistance is an effective strategy for controlling diseases. Fhb7 and Pm21 are derived from wild relativ...
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| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-07-01
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| Series: | BMC Plant Biology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12870-025-07005-8 |
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| Summary: | Abstract Background Wheat (Triticum aestivum L.) cultivation suffers from significant yield loss owing to diseases such as Fusarium head blight (FHB) and powdery mildew (PM). Utilization of host resistance is an effective strategy for controlling diseases. Fhb7 and Pm21 are derived from wild relatives of wheat, which confer broad-spectrum resistance to FHB and PM, respectively, and can be used in breeding through marker-assisted selection. Kompetitive Allele Specific polymerase chain reaction (KASP) is a homogeneous fluorescence-based technology, which identifies single nucleotide polymorphisms (SNP), and is suitable for marker-assisted selection (MAS) in large-scale breeding. However, KASP typically identifies the alleles of a single SNP, which limits the efficiency of pyramiding multiple genes during breeding. Results In this study, we developed and applied a novel multiplex KASP (Multi-KASP) system that enabled concurrent detection of two distinct genes in a single reaction. The Multi-KASP system increased the flexibility of primer requirements and differentiated genes using only basic fluorescent cassettes, thereby significantly enhancing genotyping efficiency while ensuring high specificity and accuracy. This system accurately distinguished homozygous and heterozygous genotypes for both genes, which was validated by comparison with conventional marker detection. Phenotyping showed that polymerization of Fhb7 and Pm21 using the multi-KASP system enhanced resistance to FHB and PM in the wheat breeding program. Conclusions A cost-effective Multi-KASP genotyping system, capable of simultaneously detecting two distinct genes in a single reaction, is an efficient and convenient solution for molecular breeding. The system demonstrates high potential for enhancing the efficiency of MAS in wheat breeding for resistance to FHB and PM. |
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| ISSN: | 1471-2229 |