Genetic and genomic characterization followed by single-step genomic evaluation of withers height in German Warmblood horses

Reliability of genomic predictions is influenced by the size and genetic composition of the reference population. For German Warmblood horses, compilation of a reference population has been enabled through the cooperation of five German breeding associations. In this study, preliminary data from this joint reference population were used to genetically and genomically characterize withers height and to apply single-step methodology for estimating genomic breeding values for withers height. Using data on 2113 mares and their genomic information considering about 62,000 single nucleotide polymorphisms (SNPs), analysis of the genomic relationship revealed substructures reflecting breed origin and different breeding goals of the contributing breeding associations. A genome-wide association study confirmed a known quantitative trait locus (QTL) for withers height on equine chromosome (ECA) 3 close to LCORL and identified a further significant peak on ECA 1. Using a single-step approach with a combined relationship matrix, the estimated heritability for withers height was 0.31 (SE = 0.08) and the corresponding genomic breeding values ranged from − 2.94 to 2.96 cm. A mean reliability of 0.38 was realized for these breeding values. The analyses of withers height showed that compiling a reference population across breeds is a suitable strategy for German Warmblood horses. The single-step method is an appealing approach for practical genomic prediction in horses, because not many genotypes are available yet and animals without genotypes can by this way directly contribute to the estimation system.

Genomic prediction using low-coverage portable Nanopore sequencing

Most traits in livestock, crops and humans are polygenic, that is, a large number of loci contribute to genetic variation. Effects at these loci lie along a continuum ranging from common low-effect to rare high-effect variants that cumulatively contribute to the overall phenotype. Statistical methods to calculate the effect of these loci have been developed and can be used to predict phenotypes in new individuals. In agriculture, these methods are used to select superior individuals using genomic breeding values; in humans these methods are used to quantitatively measure an individual’s disease risk, termed polygenic risk scores. Both fields typically use SNP array genotypes for the analysis. Recently, genotyping-by-sequencing has become popular, due to lower cost and greater genome coverage (including structural variants). Oxford Nanopore Technologies’ (ONT) portable sequencers have the potential to combine the benefits genotyping-by-sequencing with portability and decreased turn-around time. This introduces the potential for in-house clinical genetic disease risk screening in humans or calculating genomic breeding values on-farm in agriculture. Here we demonstrate the potential of the later by calculating genomic breeding values for four traits in cattle using low-coverage ONT sequence data and comparing these breeding values to breeding values calculated from SNP arrays. At sequencing coverages between 2X and 4X the correlation between ONT breeding values and SNP array-based breeding values was > 0.92 when imputation was used and > 0.88 when no imputation was used. With an average sequencing coverage of 0.5x the correlation between the two methods was between 0.85 and 0.92 using imputation, depending on the trait. This suggests that ONT sequencing has potential for in clinic or on-farm genomic prediction, however, further work to validate these findings in a larger population still remains.

43 Single and Multiple-breed Genomic Predictions for Conformation Traits of Canadian Dairy Goats

The use of multiple-breed models can increase the accuracy of estimated breeding values (EBV) when few phenotypes are available for a trait. However, pooling breeds is not always beneficial for genomic evaluations due to the low consistency of gametic phase between individual breeds. The objective of this study was to compare the expected gain in accuracy of single-step genomic breeding values (GEBV) for conformation traits of Canadian Alpine and Saanen goats predicted using single and multiple-breed models. The traits considered were body capacity, dairy character, feet and legs, fore udder, general appearance, rear udder, suspensory ligament, and teats, all recorded by trained classifiers, using a 1 to 9 scale. The full datasets included a total of 7,500 phenotypes for each trait (5,158 Alpine and 2,342 Saanen) and 1,707 50K genotypes (833 Alpine, 874 Saanen). Standard errors of prediction (SEP) were obtained for EBV and GEBV predicted using single-trait animal models on full or validation datasets. Breed difference was accounted for as a fixed effect in the multiple-breed models. Average theoretical accuracies were calculated from the SEP. For Saanen, with fewer records, expected accuracies of EBV and GEBV for the validation animals (selection candidates) were consistently higher for the multiple-breed models. Trait specific gains in theoretical accuracy of GEBV relative to EBV for the selection candidates ranged from 30 to 48% for Alpine and 41 to 61% for Saanen. Averaged across all traits, GEBV predicted from the full dataset were 32 to 38% more accurate than EBV for genotyped animals and the largest gains were found for does without conformation records (49 to 55%) and bucks without daughter records (56 to 82%). Overall, the implementation of genomic selection would substantially increase selection accuracy for young breeding candidates and, consequently, the rate of genetic improvement for conformation traits in Canadian dairy goats.

37 Single-step Genomic BLUP Fitting Snps or Haplotypes in Genetically-diverse Populations: A Simulation Study

The successful implementation of genomic selection in more genetically diverse populations (e.g., sheep and goats) require larger training populations. Haplotype-based genomic predictions are hypothesized to perform better in comparison to single-SNP methods mainly due to the possibility of better capturing QTL effects in linkage disequilibrium (LD) with the markers. However, most genomic-prediction studies based on haplotypes were performed in populations with low effective population size (Ne < 150). We aimed to investigate alternative approaches for fitting haplotypes using the single-step GBLUP method (ssGBLUP) in a genetically diverse population (Ne = 400). We simulated a composite sheep population, mimicking real populations based on literature parameters, using the QMSim software, with five replicates. We simulated a HD panel (600K) and two traits with different heritabilites (0.10 and 0.30). Pseudo-SNPs from unique haplotype alleles derived from LD blocks with thresholds of 0.1, 0.3, and 0.6 (LD01, LD03, and LD06, respectively) were used in the analyses. The LD-blocks were constructed using a 50K panel designed from the simulated HD. The training population was composed of 60,000 individuals with phenotypes, 8,000 of them also had genotypes, and 2,000 young genotyped individuals were used as the validation set. The genomic relationship (G) in the ssGBLUP was constructed using both independent markers and pseudo-SNPs (haplotypes). A linear mixed effects model was used to test the effect of the G on the accuracies of prediction, followed by the Tukey test with 5% of significance. No blocks were created with LD06. The accuracies with the 50K panel, LD01, and LD03 for the moderate heritability were 0.41(0.00), 0.40(0.01) and 0.41(0.00), respectively, and 0.24(0.01) 0.23(0.01), and 0.24(0.01) for the low heritability scenario. No statistical differences were observed. Based on our findings, haplotype-based predictions did not improve the accuracy of genomic breeding values in genetically diverse populations.

The identification of grain size genes by RapMap reveals directional selection during rice domestication

Cloning quantitative trait locus (QTL) is time consuming and laborious, which hinders the understanding of natural variation and genetic diversity. Here, we introduce RapMap, a method for rapid multi-QTL mapping by employing F2 gradient populations (F2GPs) constructed by minor-phenotypic-difference accessions. The co-segregation standard of the single-locus genetic models ensures simultaneous integration of a three-in-one framework in RapMap i.e. detecting a real QTL, confirming its effect, and obtaining its near-isogenic line-like line (NIL-LL). We demonstrate the feasibility of RapMap by cloning eight rice grain-size genes using 15 F2GPs in three years. These genes explain a total of 75% of grain shape variation. Allele frequency analysis of these genes using a large germplasm collection reveals directional selection of the slender and long grains in indica rice domestication. In addition, major grain-size genes have been strongly selected during rice domestication. We think application of RapMap in crops will accelerate gene discovery and genomic breeding.

The Bnapus50K array: a quick and versatile genotyping tool for Brassica napus genomic breeding and research

Rapeseed is a globally cultivated commercial crop, primarily grown for its oil. High-density single nucleotide polymorphism (SNP) arrays are widely used as a standard genotyping tool for rapeseed research, including for gene mapping, genome-wide association studies, germplasm resource analysis, and cluster analysis. Although considerable rapeseed genome sequencing data has been released, DNA arrays are still an attractive choice for providing additional genetic data in an era of high-throughput whole-genome sequencing. Here, we integrated re-sequencing DNA array data (32,216, and 304 SNPs) from 505 inbred rapeseed lines, allowing us to develop a sensitive and efficient genotyping DNA array, Bnapus50K, with a more consistent genetic and physical distribution of probes. A total of 42,090 high quality probes were filtered and synthesized, with an average distance between adjacent SNPs of 8 kb. To improve the practical application potential of this array in rapeseed breeding, we also added 1,618 functional probes related to important agronomic traits such as oil content, disease resistance, male sterility, and flowering time. The additional probes also included those specifically for detecting genetically modified material. These probes show a good detection efficiency and are therefore useful for gene mapping, along with crop variety improvement and identification. The novel Bnapus50K DNA array developed in this study could prove to be a quick and versatile genotyping tool for B. napus genomic breeding and research.