A compendium of genetic regulatory effects across pig tissues.

The Farm Animal Genotype-Tissue Expression (FarmGTEx) project has been established to develop a public resource of genetic regulatory variants in livestock, which is essential for linking genetic polymorphisms to variation in phenotypes, helping fundamental biological discovery and exploitation in animal breeding and human biomedicine. Here we show results from the pilot phase of PigGTEx by processing 5,457 RNA-sequencing and 1,602 whole-genome sequencing samples passing quality control from pigs. We build a pig genotype imputation panel and associate millions of genetic variants with five types of transcriptomic phenotypes in 34 tissues. We evaluate tissue specificity of regulatory effects and elucidate molecular mechanisms of their action using multi-omics data. Leveraging this resource, we decipher regulatory mechanisms underlying 207 pig complex phenotypes and demonstrate the similarity of pigs to humans in gene expression and the genetic regulation behind complex phenotypes, supporting the importance of pigs as a human biomedical model.

PigBiobank: a valuable resource for understanding genetic and biological mechanisms of diverse complex traits in pigs.

To fully unlock the potential of pigs as both agricultural species for animal-based protein food and biomedical models for human biology and disease, a comprehensive understanding of molecular and cellular mechanisms underlying various complex phenotypes in pigs and how the findings can be translated to other species, especially humans, are urgently needed. Here, within the Farm animal Genotype-Tissue Expression (FarmGTEx) project, we build the PigBiobank (http://pigbiobank.farmgtex.org) to systematically investigate the relationships among genomic variants, regulatory elements, genes, molecular networks, tissues and complex traits in pigs. This first version of the PigBiobank curates 71 885 pigs with both genotypes and phenotypes from over 100 pig breeds worldwide, covering 264 distinct complex traits. The PigBiobank has the following functions: (i) imputed sequence-based genotype-phenotype associations via a standardized and uniform pipeline, (ii) molecular and cellular mechanisms underlying trait-associations via integrating multi-omics data, (iii) cross-species gene mapping of complex traits via transcriptome-wide association studies, and (iv) high-quality results display and visualization. The PigBiobank will be updated timely with the development of the FarmGTEx-PigGTEx project, serving as an open-access and easy-to-use resource for genetically and biologically dissecting complex traits in pigs and translating the findings to other species.

Preselecting Variants from Large-Scale Genome-Wide Association Study Meta-Analyses Increases the Genomic Prediction Accuracy of Growth and Carcass Traits in Large White Pigs.

Preselected variants associated with the trait of interest from genome-wide association studies (GWASs) are available to improve genomic prediction in pigs. The objectives of this study were to use preselected variants from a large GWAS meta-analysis to assess the impact of single-nucleotide polymorphism (SNP) preselection strategies on genome prediction of growth and carcass traits in pigs. We genotyped 1018 Large White pigs using medium (50k) SNP arrays and then imputed SNPs to sequence level by utilizing a reference panel of 1602 whole-genome sequencing samples. We tested the effects of different proportions of selected top SNPs across different SNP preselection strategies on genomic prediction. Finally, we compared the prediction accuracies by employing genomic best linear unbiased prediction (GBLUP), genomic feature BLUP and three weighted GBLUP models. SNP preselection strategies showed an average improvement in accuracy ranging from 0.3 to 2% in comparison to the SNP chip data. The accuracy of genomic prediction exhibited a pattern of initial increase followed by decrease, or continuous decrease across various SNP preselection strategies, as the proportion of selected top SNPs increased. The highest level of prediction accuracy was observed when utilizing 1 or 5% of top SNPs. Compared with the GBLUP model, the utilization of estimated marker effects from a GWAS meta-analysis as SNP weights in the BLUP|GA model improved the accuracy of genomic prediction in different SNP preselection strategies. The new SNP preselection strategies gained from this study bring opportunities for genomic prediction in limited-size populations in pigs.

Integration of non-additive genome-wide association study with a multi-tissue transcriptome analysis of growth and carcass traits in Duroc pigs.

Growth and carcass traits are of economic importance in the pig production, which affect pork quality and profitability of finishing pig production. This study used whole-genome and transcriptome sequencing technologies to identify potential candidate genes affecting growth and carcass traits in Duroc pigs. The medium (50-60 k) single nucleotide polymorphism (SNP) arrays of 4 154 Duroc pigs from three populations were imputed to whole-genome sequence data, yielding 10 463 227 markers on 18 autosomes. The dominance heritabilities estimated for growth and carcass traits ranged from 0.000 ± 0.041 to 0.161 ± 0.054. Using non-additive genome-wide association study (GWAS), we identified 80 dominance quantitative trait loci for growth and carcass traits at genome-wide significance (false discovery rate < 5%), 15 of which were also detected in our additive GWAS. After fine mapping, 31 candidate genes for dominance GWAS were annotated, and 8 of them were highlighted that have been previously reported to be associated with growth and development (e.g. SNX14, RELN and ENPP2), autosomal recessive diseases (e.g. AMPH, SNX14, RELN and CACNB4) and immune response (e.g. UNC93B1 and PPM1D). By integrating the lead SNPs with RNA-seq data of 34 pig tissues from the Pig Genotype-Tissue Expression project (https://piggtex.farmgtex.org/), we found that the rs691128548, rs333063869, and rs1110730611 have significantly dominant effects for the expression of SNX14, AMPH and UNC93B1 genes in tissues related to growth and development for pig, respectively. Finally, the identified candidate genes were significantly enriched for biological processes involved in the cell and organ development, lipids catabolic process and phosphatidylinositol 3-kinase signalling (P < 0.05). These results provide new molecular markers for meat production and quality selection of pig as well as basis for deciphering the genetic mechanisms of growth and carcass traits.

Evaluation of six machine learning classification algorithms in pig breed identification using SNPs array data.

Breed identification utilizing multiple information sources and methods is widely applicated in the field of animal genetics and breeding. Simultaneously, with the development of artificial intelligence, the integration of high-throughput genomic data and machine learning techniques is increasingly used for breed identification. In this context, we used 654 individuals from 15 pig breeds, evaluating the performance of machine learning and stacking ensemble learning classifiers, as well as the function of feature selection and anomaly detection in different scenarios. Our results showed that, when using a training set of 16 individuals per breed and 32 features (SNPs), the accuracy of breed identification with feature selection (eXtreme Gradient Boosting, XGBoost) could exceed 95.00% (nine breeds), and was improved by 7.04% over the results with random selection. For stacking ensemble learning, feature selection methods (including random selection method) were used before different base learners. When these base learners' training set had 16 individuals per breed and 32 features, the accuracy of stacking ensemble learning improved by 9.24% over the best base learner (nine breeds), but did not significantly increase the advantage over the models with XGBoost feature selection. When using a training set of 16 individuals and 512 features per breed, breed identification with anomaly detection (local outlier factor, LOF) and random selection could achieve an accuracy of 89.06% (15 breeds). These results show that machine learning could be an effective tool for breed identification and this study will also provide useful information for the application of machine learning in animal genetics and breeding.

Model Comparison of Heritability Enrichment Analysis in Livestock Population.

Heritability enrichment analysis is an important means of exploring the genetic architecture of complex traits in human genetics. Heritability enrichment is typically defined as the proportion of an SNP subset explained heritability, divided by the proportion of SNPs. Heritability enrichment enables better study of underlying complex traits, such as functional variant/gene subsets, biological networks and metabolic pathways detected through integrating explosively increased omics data. This would be beneficial for genomic prediction of disease risk in humans and genetic values estimation of important economical traits in livestock and plant species. However, in livestock, factors affecting the heritability enrichment estimation of complex traits have not been examined. Previous studies on humans reported that the frequencies, effect sizes, and levels of linkage disequilibrium (LD) of underlying causal variants (CVs) would affect the heritability enrichment estimation. Therefore, the distribution of heritability across the genome should be fully considered to obtain the unbiased estimation of heritability enrichment. To explore the performance of different heritability enrichment models in livestock populations, we used the VanRaden, GCTA and α models, assuming different α values, and the LDAK model, considering LD weight. We simulated three types of phenotypes, with CVs from various minor allele frequency (MAF) ranges: genome-wide (0.005 ≤ MAF ≤ 0.5), common (0.05 ≤ MAF ≤ 0.5), and uncommon (0.01 ≤ MAF < 0.05). The performances of the models with two different subsets (one of which contained known CVs and the other consisting of randomly selected markers) were compared to verify the accuracy of heritability enrichment estimation of functional variant sets. Our results showed that models with known CV subsets provided more robust enrichment estimation. Models with different α values tended to provide stable and accurate estimates for common and genome-wide CVs (relative deviation 0.5−2.2%), while tending to underestimate the enrichment of uncommon CVs. As the α value increased, enrichments from 15.73% higher than true value (i.e., 3.00) to 48.93% lower than true value for uncommon CVs were observed. In addition, the long-range LD windows (e.g., 5000 kb) led to large bias of the enrichment estimations for both common and uncommon CVs. Overall, heritability enrichment estimations were sensitive for the α value assumption and LD weight consideration of different models. Accuracy would be greatly improved by using a suitable model. This study would be helpful in understanding the genetic architecture of complex traits and provides a reference for genetic analysis in the livestock population.

Genomic Prediction Using LD-Based Haplotypes in Combined Pig Populations.

The size of reference population is an important factor affecting genomic prediction. Thus, combining different populations in genomic prediction is an attractive way to improve prediction ability. However, combining multireference population roughly cannot increase the prediction accuracy as well as expected in pig. This may be due to different linkage disequilibrium (LD) pattern differences between population. In this study, we used the imputed whole-genome sequencing (WGS) data to construct LD-based haplotypes for genomic prediction in combined population to explore the impact of different single-nucleotide polymorphism (SNP) densities, variant representation (SNPs or haplotype alleles), and reference population size on the prediction accuracy for reproduction traits. Our results showed that genomic best linear unbiased prediction (GBLUP) using the WGS data can improve prediction accuracy in multi-population but not within-population. Not only the genomic prediction accuracy of the haplotype method using 80 K chip data in multi-population but also GBLUP for the multi-population (3.4-5.9%) was higher than that within-population (1.2-4.3%). More importantly, we have found that using the haplotype method based on the WGS data in multi-population has better genomic prediction performance, and our results showed that building haploblock in this scenario based on low LD threshold (r 2 = 0.2-0.3) produced an optimal set of variables for reproduction traits in Yorkshire pig population. Our results suggested that whether the use of the haplotype method based on the chip data or GBLUP (individual SNP method) based on the WGS data were beneficial for genomic prediction in multi-population, while simultaneously combining the haplotype method and WGS data was a better strategy for multi-population genomic evaluation.

Impact of linkage disequilibrium heterogeneity along the genome on genomic prediction and heritability estimation.

BACKGROUND: Compared to medium-density single nucleotide polymorphism (SNP) data, high-density SNP data contain abundant genetic variants and provide more information for the genetic evaluation of livestock, but it has been shown that they do not confer any advantage for genomic prediction and heritability estimation. One possible reason is the uneven distribution of the linkage disequilibrium (LD) along the genome, i.e., LD heterogeneity among regions. The aim of this study was to effectively use genome-wide SNP data for genomic prediction and heritability estimation by using models that control LD heterogeneity among regions. METHODS: The LD-adjusted kinship (LDAK) and LD-stratified multicomponent (LDS) models were used to control LD heterogeneity among regions and were compared with the classical model that has no such control. Simulated and real traits of 2000 dairy cattle individuals with imputed high-density (770K) SNP data were used. Five types of phenotypes were simulated, which were controlled by very strongly, strongly, moderately, weakly and very weakly tagged causal variants, respectively. The performances of the models with high- and medium-density (50K) panels were compared to verify that the models that controlled LD heterogeneity among regions were more effective with high-density data. RESULTS: Compared to the medium-density panel, the use of the high-density panel did not improve and even decreased prediction accuracies and heritability estimates from the classical model for both simulated and real traits. Compared to the classical model, LDS effectively improved the accuracy of genomic predictions and unbiasedness of heritability estimates, regardless of the genetic architecture of the trait. LDAK applies only to traits that are mainly controlled by weakly tagged causal variants, but is still less effective than LDS for this type of trait. Compared with the classical model, LDS improved prediction accuracy by about 13% for simulated phenotypes and by 0.3 to ~ 10.7% for real traits with the high-density panel, and by ~ 1% for simulated phenotypes and by - 0.1 to ~ 6.9% for real traits with the medium-density panel. CONCLUSIONS: Grouping SNPs based on regional LD to construct the LD-stratified multicomponent model can effectively eliminate the adverse effects of LD heterogeneity among regions, and greatly improve the efficiency of high-density SNP data for genomic prediction and heritability estimation.

Utilization Strategies of Two Environment Phenotypes in Genomic Prediction.

Multiple environment phenotypes may be utilized to implement genomic prediction in plant breeding, while it is unclear about optimal utilization strategies according to its different availability. It is necessary to assess the utilization strategies of genomic prediction models based on different availability of multiple environment phenotypes. Here, we compared the prediction accuracy of three genomic prediction models (genomic prediction model (genomic best linear unbiased prediction (GBLUP), genomic best linear unbiased prediction (GFBLUP), and multi-trait genomic best linear unbiased prediction (mtGBLUP)) which leveraged diverse information from multiple environment phenotypes using a rice dataset containing 19 agronomic traits in two disparate seasons. We found that the prediction accuracy of genomic prediction models considering multiple environment phenotypes (GFBLUP and mtGBLUP) was better than the classical genomic prediction model (GBLUP model). The deviation of prediction accuracy of between GBLUP and mtGBLUP or GFBLUP was associated with the phenotypic correlation. In summary, the genomic prediction models considering multiple environment phenotypes (GFBLUP and mtGBLUP) demonstrated better prediction accuracy. In addition, we could utilize different genomic prediction strategies according to different availability of multiple environment phenotypes.