Journal
PLANT JOURNAL
Volume 113, Issue 1, Pages 26-46Publisher
WILEY
DOI: 10.1111/tpj.16028
Keywords
genotyping; genome-wide association study; pangenome; Oryza sativa; quantitative trait loci mapping; single-nucleotide polymorphism array
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The advent of the pangenome era has uncovered previously unknown genetic variation in diverse crop plants, but conventional single-reference genotyping methods often fail to capture a large portion of this variation, leading to reference bias. To overcome this, a Rice Pangenome Genotyping Array (RPGA) was developed, enabling rapid genotyping based on the entire 3K rice pangenome. This array provides a cost-effective solution and has been successfully applied in various areas, such as genome-wide association studies, population structure analysis, and marker-assisted selection.
The advent of the pangenome era has unraveled previously unknown genetic variation existing within diverse crop plants, including rice. This untapped genetic variation is believed to account for a major portion of phenotypic variation existing in crop plants. However, the use of conventional single reference-guided genotyping often fails to capture a large portion of this genetic variation leading to a reference bias. This makes it difficult to identify and utilize novel population/cultivar-specific genes for crop improvement. Thus, we developed a Rice Pangenome Genotyping Array (RPGA) harboring probes assaying 80K single-nucleotide polymorphisms (SNPs) and presence-absence variants spanning the entire 3K rice pangenome. This array provides a simple, user-friendly and cost-effective (60-80 USD per sample) solution for rapid pangenome-based genotyping in rice. The genome-wide association study (GWAS) conducted using RPGA-SNP genotyping data of a rice diversity panel detected a total of 42 loci, including previously known as well as novel genomic loci regulating grain size/weight traits in rice. Eight of these identified trait-associated loci (dispensable loci) could not be detected with conventional single reference genome-based GWAS. A WD repeat-containing PROTEIN 12 gene underlying one of such dispensable locus on chromosome 7 (qLWR7) along with other non-dispensable loci were subsequently detected using high-resolution quantitative trait loci mapping confirming authenticity of RPGA-led GWAS. This demonstrates the potential of RPGA-based genotyping to overcome reference bias. The application of RPGA-based genotyping for population structure analysis, hybridity testing, ultra-high-density genetic map construction and chromosome-level genome assembly, and marker-assisted selection was also demonstrated. A web application () was further developed to provide an easy to use platform for the imputation of RPGA-based genotyping data using 3K rice reference panel and subsequent GWAS.
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