Abstract
SummaryFor 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.
Highlights
Bread wheat (Triticum aestivum L.) is one of the most widely grown crops in the world and accounts for almost one-fifth of the human calorie intake (FAO, 2017)
We focused on single nucleotide polymorphism (SNP)-containing probes on the array that potentially had a single-copy in the wheat genome
This study has described the design, validation and implementation of chromosome-specific KASP markers in wheat
Summary
Bread wheat (Triticum aestivum L.) is one of the most widely grown crops in the world and accounts for almost one-fifth of the human calorie intake (FAO, 2017). Recently called ‘introgressiomics’ (Prohens et al, 2017), consists of a whole-genome introgression approach involving transfer of chromosome segments from the entire genome of a wild relative species into the wheat background, irrespective of any traits that the wild relative might carry and a number of such studies have already been undertaken (Grewal et al, 2018a,b; King et al., 2017, 2018; Valkoun, 2001) In this prebreeding strategy, the interspecific hybrids are repeatedly backcrossed to the elite wheat parent to reduce the number and size of the introgressed segments and self-fertilized to obtain stable homozygous introgressions that can be utilized for trait analysis (King et al, 2019). Molecular markers provide high-throughput and cost-effective evaluation of introgressions in large numbers of lines (Thomson, 2014)
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