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Rapid identification of homozygosity and site of wild relative introgressions in wheat through chromosome-specific KASP genotyping assays

Grewal, Surbhi; Hubbart-Edwards, Stella; Yang, Caiyun; Devi, Urmila; Baker, Lauren; Heath, Jack; Ashling, Stephen; Scholefield, Duncan; Howells, Caroline; Yarde, Jermaine; Isaac, Peter; King, Ian P.; King, Julie


Caiyun Yang

Urmila Devi

Lauren Baker

Jack Heath

Stephen Ashling

Duncan Scholefield

Caroline Howells

Jermaine Yarde

Peter Isaac


For future food security, it is important that wheat, one of the most widely consumed crops in the world, can survive the threat of abiotic and biotic stresses. New genetic variation is currently being introduced into wheat through introgressions from its wild relatives. For trait discovery, it is necessary that each introgression is homozygous and hence stable. Breeding programmes rely on efficient genotyping platforms for marker‐assisted selection (MAS). Recently, single nucleotide polymorphism (SNP)‐based markers have been made available on high‐throughput Axiom® SNP genotyping arrays. However, these arrays are inflexible in their design and sample numbers, making their use unsuitable for long‐term MAS. SNPs can potentially be converted into Kompetitive allele‐specific PCR (KASP™) assays that are comparatively cost‐effective and efficient for low‐density genotyping of introgression lines. However, due to the polyploid nature of wheat, KASP assays for homoeologous SNPs can have difficulty in distinguishing between heterozygous and homozygous hybrid lines in a backcross population. To identify co‐dominant SNPs, that can differentiate between heterozygotes and homozygotes, we PCR‐amplified and sequenced genomic DNA from potential single‐copy regions of the wheat genome and compared them to orthologous copies from different wild relatives. A panel of 620 chromosome‐specific KASP assays have been developed that allow rapid detection of wild relative segments and provide information on their homozygosity and site of introgression in the wheat genome. A set of 90 chromosome‐nonspecific assays was also produced that can be used for genotyping introgression lines. These multipurpose KASP assays represent a powerful tool for wheat breeders worldwide.


Grewal, S., Hubbart-Edwards, S., Yang, C., Devi, U., Baker, L., Heath, J., …King, J. (2020). Rapid identification of homozygosity and site of wild relative introgressions in wheat through chromosome-specific KASP genotyping assays. Plant Biotechnology Journal, 18(3), 743-755.

Journal Article Type Article
Acceptance Date Aug 17, 2019
Online Publication Date Aug 29, 2019
Publication Date 2020-03
Deposit Date Oct 16, 2019
Publicly Available Date Feb 7, 2020
Journal Plant Biotechnology Journal
Print ISSN 1467-7644
Electronic ISSN 1467-7652
Publisher Wiley
Peer Reviewed Peer Reviewed
Volume 18
Issue 3
Pages 743-755
Public URL
Publisher URL