Zhiyan Zhang

Marker imputation with low-density marker panels in Dutch Holstein cattle

The availability of high-density bovine genotyping arrays made implementation of genomic selection possible in dairy cattle. Development of low-density single nucleotide polymorphism (SNP) panels will allow the extension of genomic selection to a larger portion of the population. Prediction of ungenotyped markers, called imputation, is a strategy that allows using the same low-density chips for all traits (and for different breeds). In the present study, we evaluated the accuracy of imputation with low-density genotyping arrays in the Dutch Holstein population. Five different sizes of genotyping arrays were tested, from 384 to 6,000 SNP. According to marker density, the overall allelic imputation error rate obtained with the program DAGPHASE, which relies on linkage disequilibrium and linkage, ranged from 11.7 to 2.0%, and that obtained with the program CHROMIBD, which relies on linkage and the set of all genotyped ancestors, ranged from 10.7 to 3.3%. However, imputation efficiency was influenced by the relationship between low-density and high-density genotyped animals. Animals with both parents genotyped had particularly low imputation error rates: <1% with 1,500 SNP or more. In summary, missing marker alleles can be predicted with 3 to 4% errors with approximately 1 SNP/Mb (approximately 3,000 markers). The CHROMIBD program proved more efficient than DAGPHASE only at lower marker densities or when several genotyped ancestors were available. Future studies are required to measure the effect of these imputation error rates on accuracy of genomic selection with low-density SNP panels.

Serial translocation by means of circular intermediates underlies colour sidedness in cattle

Colour sidedness is a dominantly inherited phenotype of cattle characterized by the polarization of pigmented sectors on the flanks, snout and ear tips. It is also referred to as ‘lineback’ or ‘witrik’ (which means white back), as colour-sided animals typically display a white band along their spine. Colour sidedness is documented at least since the Middle Ages and is presently segregating in several cattle breeds around the globe, including in Belgian blue and brown Swiss. Here we report that colour sidedness is determined by a first allele on chromosome 29 (Cs29), which results from the translocation of a 492-kilobase chromosome 6 segment encompassing KIT to chromosome 29, and a second allele on chromosome 6 (Cs6), derived from the first by repatriation of fused 575-kilobase chromosome 6 and 29 sequences to the KIT locus. We provide evidence that both translocation events involved circular intermediates. This is the first example, to our knowledge, of a phenotype determined by homologous yet non-syntenic alleles that result from a novel copy-number-variant-generating mechanism.

A splice site variant in the bovine RNF11 gene compromises growth and regulation of the inflammatory response.

We report association mapping of a locus on bovine chromosome 3 that underlies a Mendelian form of stunted growth in Belgian Blue Cattle (BBC). By resequencing positional candidates, we identify the causative c124-2A>G splice variant in intron 1 of the RNF11 gene, for which all affected animals are homozygous. We make the remarkable observation that 26% of healthy Belgian Blue animals carry the corresponding variant. We demonstrate in a prospective study design that approximately one third of homozygous mutants die prematurely with major inflammatory lesions, hence explaining the rarity of growth-stunted animals despite the high frequency of carriers. We provide preliminary evidence that heterozygous advantage for an as of yet unidentified phenotype may have caused a selective sweep accounting for the high frequency of the RNF11 c124-2A>G mutation in Belgian Blue Cattle.

Ancestral haplotype-based association mapping with generalized linear mixed models accounting for stratification

MOTIVATION In many situations, genome-wide association studies are performed in populations presenting stratification. Mixed models including a kinship matrix accounting for genetic relatedness among individuals have been shown to correct for population and/or family structure. Here we extend this methodology to generalized linear mixed models which properly model data under various distributions. In addition we perform association with ancestral haplotypes inferred using a hidden Markov model. RESULTS The method was shown to properly account for stratification under various simulated scenari presenting population and/or family structure. Use of ancestral haplotypes resulted in higher power than SNPs on simulated datasets. Application to real data demonstrates the usefulness of the developed model. Full analysis of a dataset with 4600 individuals and 500 000 SNPs was performed in 2 h 36 min and required 2.28 Gb of RAM. AVAILABILITY The software GLASCOW can be freely downloaded from www.giga.ulg.ac.be/jcms/prod_381171/software. CONTACT francois.guillaume@jouy.inra.fr SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online.

A missense mutation accelerating the gating of the lysosomal Cl−/H+-exchanger ClC-7/Ostm1 causes osteopetrosis with gingival hamartomas in cattle

Chloride-proton exchange by the lysosomal anion transporter ClC-7/Ostm1 is of pivotal importance for the physiology of lysosomes and bone resorption. Mice lacking either ClC-7 or Ostm1 develop a lysosomal storage disease and mutations in either protein have been found to underlie osteopetrosis in mice and humans. Some human disease-causing CLCN7 mutations accelerate the usually slow voltage-dependent gating of ClC-7/Ostm1. However, it has remained unclear whether the fastened kinetics is indeed causative for the disease. Here we identified and characterized a new deleterious ClC-7 mutation in Belgian Blue cattle with a severe symptomatology including perinatal lethality and in most cases gingival hamartomas. By autozygosity mapping and genome-wide sequencing we found a handful of candidate variants, including a cluster of three private SNPs causing the substitution of a conserved tyrosine in the CBS2 domain of ClC-7 by glutamine. The case for ClC-7 was strengthened by subsequent examination of affected calves that revealed severe osteopetrosis. The Y750Q mutation largely preserved the lysosomal localization and assembly of ClC-7/Ostm1, but drastically accelerated its activation by membrane depolarization. These data provide first evidence that accelerated ClC-7/Ostm1 gating per se is deleterious, highlighting a physiological importance of the slow voltage-activation of ClC-7/Ostm1 in lysosomal function and bone resorption.

Identifying cows with subclinical mastitis by bulk single nucleotide polymorphism genotyping of tank milk

Mastitis remains the most important health issue in dairy cattle. Improved methods to identify cows developing subclinical mastitis would benefit farmers. We herein describe a novel method to determine the somatic cell counts (SCC) of individual cows by bulk genotyping a sample of milk from the milk tank with panels of genome-wide single nucleotide polymorphisms (SNP). We developed a simple linear model to estimate the contribution of individual cows to the genomic DNA present in the tank milk from 1) the known genotypes of individual cows for the interrogated SNP and 2) the ratio of SNP alleles in the tank milk. Using simulations, we estimate that 3,000, 50,000, and 700,000 SNP are sufficient to accurately (R(2)>0.98) estimate individual SCC in tanks containing milk from 25, 100, and 500 cows, respectively. Using actual data, we demonstrate that the SCC of 21 cows can be estimated with a coefficient of determination of 0.60 using approximately 9,000 SNP. The proposed method increases the value of the proposition of SNP genotyping individual cows for genomic selection purposes.