A low‐coverage skim‐sequencing and imputation pipeline for genomic selection

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Main Authors: Sajal R. Sthapit, Jared Crain, Steve Larson, James A. Anderson, Prabin Bajgain, Lee R. DeHaan, Jesse Poland
Format: Artículo Open Access
Published: Wiley 2025
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author Sajal R. Sthapit
Jared Crain
Steve Larson
James A. Anderson
Prabin Bajgain
Lee R. DeHaan
Jesse Poland
author_facet Sajal R. Sthapit
Jared Crain
Steve Larson
James A. Anderson
Prabin Bajgain
Lee R. DeHaan
Jesse Poland
Sajal R. Sthapit
Jared Crain
Steve Larson
James A. Anderson
Prabin Bajgain
Lee R. DeHaan
Jesse Poland
collection Wiley Open Access
contents A low‐coverage skim‐sequencing and imputation pipeline for genomic selection Sajal R. Sthapit Jared Crain Steve Larson James A. Anderson Prabin Bajgain Lee R. DeHaan Jesse Poland The Plant Genome Abstract Genomic selection (GS) can accelerate plant breeding gains by reducing breeding cycle times, reducing phenotyping costs, or improving selection accuracy. GS is especially promising for perennial crops such as intermediate wheatgrass (IWG, Thinopyrum intermedium ) that may require multiple years of evaluation under phenotypic recurrent selection. A major obstacle in implementing GS is the need for an affordable, high‐density, genetic marker system that is scalable to thousands or tens‐of‐thousands of samples in breeding programs, especially in emerging or minor crop species. As sequencing costs continue to decrease, low‐coverage whole genome skim‐sequencing (skim‐seq) has become an attractive method for GS. Using commercial laboratory products and open‐source software, we implemented whole genome prediction at breeding program scale using ultra‐low coverage (0.01x– 0.05x, 100–125 million reads per sample) whole genome skim‐seq. Using STITCH (Sequencing to Imputation Through Constructing Haplotypes) imputation software, we evaluated optimization of imputation parameters including sequence coverage and number of assumed ancestral haplotypes. Finally, we evaluated whole genome prediction cross‐validation accuracies using reduced representation genotyping‐by‐sequencing (GBS) versus skim‐seq data for IWG, an outcrossing, heterozygous, large genome (12.7 Gb), polyploid perennial species. Our results indicate correlations between cross‐validation accuracies across five traits in IWG using skim‐seq data ( r  = 0.29–0.61) can be used as effectively as GBS ( r  = 0.29–0.55) while generating low‐coverage archival sequence data that will be robust to technological advances. These methods will be applicable to a wide range of crops and scale to breeding program size, allowing for more tractable implementation of GS within breeding programs. 10.1002/tpg2.70139 http://creativecommons.org/licenses/by/4.0/
doi_str_mv 10.1002/tpg2.70139
format Artículo Open Access
id wiley_oa_10_1002_tpg2_70139
institution Wiley Open Access
license_str_mv http://creativecommons.org/licenses/by/4.0/
publishDate 2025
publisher Wiley
record_format wiley_oa
spellingShingle A low‐coverage skim‐sequencing and imputation pipeline for genomic selection
Sajal R. Sthapit
Jared Crain
Steve Larson
James A. Anderson
Prabin Bajgain
Lee R. DeHaan
Jesse Poland
The Plant Genome
A low‐coverage skim‐sequencing and imputation pipeline for genomic selection Sajal R. Sthapit Jared Crain Steve Larson James A. Anderson Prabin Bajgain Lee R. DeHaan Jesse Poland The Plant Genome Abstract Genomic selection (GS) can accelerate plant breeding gains by reducing breeding cycle times, reducing phenotyping costs, or improving selection accuracy. GS is especially promising for perennial crops such as intermediate wheatgrass (IWG, Thinopyrum intermedium ) that may require multiple years of evaluation under phenotypic recurrent selection. A major obstacle in implementing GS is the need for an affordable, high‐density, genetic marker system that is scalable to thousands or tens‐of‐thousands of samples in breeding programs, especially in emerging or minor crop species. As sequencing costs continue to decrease, low‐coverage whole genome skim‐sequencing (skim‐seq) has become an attractive method for GS. Using commercial laboratory products and open‐source software, we implemented whole genome prediction at breeding program scale using ultra‐low coverage (0.01x– 0.05x, 100–125 million reads per sample) whole genome skim‐seq. Using STITCH (Sequencing to Imputation Through Constructing Haplotypes) imputation software, we evaluated optimization of imputation parameters including sequence coverage and number of assumed ancestral haplotypes. Finally, we evaluated whole genome prediction cross‐validation accuracies using reduced representation genotyping‐by‐sequencing (GBS) versus skim‐seq data for IWG, an outcrossing, heterozygous, large genome (12.7 Gb), polyploid perennial species. Our results indicate correlations between cross‐validation accuracies across five traits in IWG using skim‐seq data ( r  = 0.29–0.61) can be used as effectively as GBS ( r  = 0.29–0.55) while generating low‐coverage archival sequence data that will be robust to technological advances. These methods will be applicable to a wide range of crops and scale to breeding program size, allowing for more tractable implementation of GS within breeding programs. 10.1002/tpg2.70139 http://creativecommons.org/licenses/by/4.0/
title A low‐coverage skim‐sequencing and imputation pipeline for genomic selection
topic The Plant Genome
url https://acsess.onlinelibrary.wiley.com/doi/10.1002/tpg2.70139