Isolation of subtelomeric sequences of porcine chromosomes for translocation screening reveals errors in the pig genome assembly.

Balanced chromosomal aberrations have been shown to affect fertility in most species studied, often leading to hypoprolificacy (reduced litter size) in domestic animals such as pigs. With an increasing emphasis in modern food production on the use of a small population of high quality males for artificial insemination, the potential economic and environmental costs of hypoprolific boars, bulls, rams etc. are considerable. There is therefore a need for novel tools to facilitate rapid, cost-effective chromosome translocation screening. This has previously been achieved by standard karyotype analysis; however, this approach relies on a significant level of expertise and is limited in its ability to identify subtle, cryptic translocations. To address this problem, we developed a novel device and protocol for translocation screening using subtelomeric probes and fluorescence in situ hybridisation. Probes were designed using BACs (bacterial artificial chromosomes) from the subtelomeric region of the short (p-arm) and long (q-arm) of each porcine chromosome. They were directly labelled with FITC or Texas Red (p-arm and q-arm respectively) prior to application of a 'Multiprobe' device, thereby enabling simultaneous detection of each individual porcine chromosome on a single slide. Initial experiments designed to isolate BACs in subtelomeric regions led to the discovery of a series of incorrectly mapped regions in the porcine genome assembly (from a total of 82 BACs, only 45 BACs mapped correctly). Our work therefore highlights the importance of accurate physical mapping of newly sequenced genomes. The system herein described allows for robust and comprehensive analysis of the porcine karyotype, an adjunct to classical cytogenetics that provides a valuable tool to expedite efficient, cost effective food production.

Effects of low protein diets on pigs with a lean genotype 2. Compositional traits measured with computed tomography (CT).

The effects on compositional changes across the growing-finishing period (40-115 kg) of feeding pig diets with different protein and amino acid levels were investigated using CT scanning (at 60, 85 and 115 kg live weight). Pigs of a lean commercial genotype were fed a commercial control regime (C), or a low protein regime with either high (LP1) or low (LP2) essential amino acid levels, all balanced for net energy. In vivo CT measurements agreed well with post-slaughter sample joint dissection results for carcass tissue weights/proportions, and CT-measured muscle density predicted intramuscular fat accurately. Pigs on C and LP1 regimes did not differ significantly in composition during growth. However, pigs on the LP2 regime had significantly more fat (in carcass, internal and intra-muscular depots) and less muscle, from 85 kg onwards. Although fat levels differed depending on diet regime (LP2>others), proportions of fat in different body depots were unaffected.

Effects of low protein diets on pigs with a lean genotype. 1. Carcass composition measured by dissection and muscle fatty acid composition.

Pigs with a lean genotype were fed diets differing in protein and amino acid contents between 40 and 115 kg live weight. A high protein control regime (C) was compared with one supplying 11% less total protein but the same essential amino acid levels (LP1) and one supplying 16% less protein but lower amino acid levels (LP2). Regime LP2 produced fatter pigs in terms of subcutaneous, intermuscular and intramuscular fat (IMF), the latter measured in longissimus and semimembranosus muscles. The percentage of linoleic acid was lowest and that of oleic acid highest in IMF from LP2 pigs (11.57 and 34.59% respectively in longissimus). Pigs in regime LP1 had more longissimus IMF than C but similar semimembranosus IMF although both muscles had lower percentages of linoleic acid in LP1 than C, suggesting a tendency towards greater fat deposition in LP1. The high IMF content in LP2 produced the most tender, juicy steaks.

Estimation of genetic associations between reproduction and production traits based on a sire and dam line with common ancestry.

Genetic parameters for survival, reproduction and production traits were estimated for a sire and dam line, originating from one Large White breed separated more than 25 years ago. The change in parameters due to different selection pressure on reproduction and production traits in both lines was also examined. Data collected between 1990 and 2007 were available for the analysis of reproduction traits in 4713 litters (sire line) and 14836 litters (dam line) and for the production traits in 58329 pigs (sire line) and 108912 pigs (dam line). Genetic parameters were estimated using a Bayesian approach. Average phenotypic differences between lines were substantial with 1.5 more piglets born in the dam line and 1.7 mm less backfat thickness (BF) in the sire line. Based on a multiple trait analysis which included both reproduction and production traits, heritabilities for survival and litter size traits in the sire (or dam) line were estimated at 0.03 ± 0.01 (0.06 ± 0.01) for percentage of stillborn piglets (SB), 0.10 ± 0.03 (0.11 ± 0.01) for total number of piglets born (NBT) and 0.09 ± 0.03 (0.09 ± 0.01) for number of piglets born alive. Heritabilities for production traits were estimated at 0.29 ± 0.01 (0.29 ± 0.01) for average daily gain, 0.50 ± 0.01 (0.42 ± 0.01) for BF and 0.41 ± 0.01 for muscle depth. Selection pressure on litter size in the dam line resulted in a slightly unfavourable correlation for SB-NBT (0.21 ± 0.11), which was only marginally unfavourable in the sire line (0.06 ± 0.24). Selection pressure on BF in the sire line may have resulted in the moderately undesirable correlation with SB (-0.46 ± 0.15), which was not significant in the dam line (-0.08 ± 0.06). Changing the base population in the dam line to animals born since the year 2000 indicated that selection pressure on different traits has altered the heritabilities and correlations of the traits within the line. The undesirable correlations between survival at birth and reproduction traits or production traits were low so that simultaneous improvement of all traits can be achieved. Heritabilities for survival at birth and reproduction traits were low, but genetic variation was substantial and extensive pedigree information can be used to improve the accuracy of breeding values, so that genetic improvement is expected to be efficient.

Mapping of quantitative trait loci for growth and carcass traits in commercial sheep populations.

Quantitative trait loci analyses were applied to data from Suffolk and Texel commercial sheep flocks in the United Kingdom. The populations comprised 489 Suffolk animals in three half-sib families and 903 Texel animals in nine half-sib families. Phenotypic data comprised measurements of live weight at 8 and 20 wk of age and ultrasonically measured fat and muscle depth at 20 wk. Lambs and their sires were genotyped across candidate regions on chromosomes 1, 2, 3, 4, 5, 6, 11, 18, and 20. Data were analyzed at the breed level, at the family level, and across extended families when families were genetically related. The breed-level analyses revealed a suggestive QTL on chromosome 1 in the Suffolk breed, between markers BM8246 and McM130, affecting muscle depth, although the effect was only significant in one of the three Suffolk families. A two-QTL analysis suggested that this effect may be due to two adjacent QTL acting in coupling. In total, 24 suggestive QTL were identified from individual family analyses. The most significant QTL affected fat depth and was segregating in a Texel family on chromosome 2, with an effect of 0.62 mm. The QTL was located around marker ILSTS030, 26 cM distal to myostatin. Two of the Suffolk and two of the Texel sires were related, and a three-generation analysis was applied across these two extended families. Seven suggestive QTL were identified in this analysis, including one that had not been detected in the individual family analysis. The most significant QTL, which affected muscle depth, was located on chromosome 18 near the callipyge and Carwell loci. Based on the phenotypic effect and location of the QTL, the data suggest that a locus similar to the Carwell locus may be segregating in the United Kingdom Texel population.

Increased heterozygosity and allele variants are seen in Texel compared to Suffolk sheep.

In this study, the Suffolk and Texel sheep breeds were compared for microsatellite marker heterozygosity throughout seven chromosomal regions in the sheep genome. A total of 623 Texel animals and 489 Suffolk animals in five and three half-sib families, respectively, were genotyped for microsatellite markers across the seven different chromosomes. Using the observed allele frequencies, the expected levels of heterozygosity were calculated for each family. The expected levels of heterozygosity did not significantly differ between the breeds across all regions studied. However, levels of expected heterozygosity were 32% higher in Texel animals on chromosome 4 due to a region of increased heterozygosity between BMS648 and BM3212. The number of allelic variants significantly differed between the breeds, solely due to a region of increased number of alleles on chromosome 20. This region of higher numbers of allele variants in the Texel breed extended from the MHC to c. 15 cM distal to the MHC region incorporating markers OMHC1, CSRD226, TGLA387 and BM1818, which had 3.30, 7.02, 3.09 and 6.75 more alleles in Texel than in Suffolk animals, respectively. No difference was observed in the variance of allele frequency between the two breeds. It is proposed that previous selective sweeps may have reduced numbers of alleles and levels of heterozygosity in the Suffolk breed.

Highly prolific Booroola sheep have a mutation in the intracellular kinase domain of bone morphogenetic protein IB receptor (ALK-6) that is expressed in both oocytes and granulosa cells.

The Booroola fecundity gene (FecB) increases ovulation rate and litter size in sheep and is inherited as a single autosomal locus. The effect of FecB is additive for ovulation rate (increasing by about 1.6 corpora lutea per cycle for each copy) and has been mapped to sheep chromosome 6q23-31, which is syntenic to human chromosome 4q21-25. Bone morphogenetic protein IB (BMP-IB) receptor (also known as ALK-6), which binds members of the transforming growth factor-beta (TGF-beta) superfamily, is located in the region containing the FecB locus. Booroola sheep have a mutation (Q249R) in the highly conserved intracellular kinase signaling domain of the BMP-IB receptor. The mutation segregated with the FecB phenotype in the Booroola backcross and half-sib flocks of sheep with no recombinants. The mutation was not found in individuals from a number of sheep breeds not derived from the Booroola strain. BMPR-IB was expressed in the ovary and in situ hybridization revealed its specific location to the oocyte and the granulosa cell. Expression of mRNA encoding the BMP type II receptor was widespread throughout the ovary. The mutation in BMPR-IB found in Booroola sheep is the second reported defect in a gene from the TGF-beta pathway affecting fertility in sheep following the recent discovery of mutations in the growth factor, GDF9b/BMP15.

Combined analyses of data from quantitative trait loci mapping studies. Chromosome 4 effects on porcine growth and fatness.

For many species several similar QTL mapping populations have been produced and analyzed independently. Joint analysis of such data could be used to increase power to detect QTL and evaluate population differences. In this study, data were collated on almost 3000 pigs from seven different F(2) crosses between Western commercial breeds and either the European wild boar or the Chinese Meishan breed. Genotypes were available for 31 markers on chromosome 4 (on average 8.3 markers per population). Data from three traits common to all populations (birth weight, mean backfat depth at slaughter or end of test, and growth rate from birth to slaughter or end of test) were analyzed for individual populations and jointly. A QTL influencing birth weight was detected in one individual population and in the combined data, with no significant interaction of the QTL effect with population. A QTL affecting backfat that had a significantly greater effect in wild boar than in Meishan crosses was detected. Some evidence for a QTL affecting growth rate was detected in all populations, with no significant differences between populations. This study is the largest F(2) QTL analysis achieved in a livestock species and demonstrates the potential of joint analysis.

Mapping of quantitative trait loci on porcine chromosome 4.

A F2 population derived from a cross between European Large White and Chinese Meishan pigs was established in order to study the genetic basis of breed differences for growth and fat traits. Chromosome 4 was chosen for initial study as previous work had revealed quantitative trait loci (QTLs) on this chromosome affected growth and fat traits in a Wild Boar x Large White cross. Individuals in the F2 population were typed for nine markers spanning a region of approximately 124 CM. We found evidence for QTLs affecting growth between weaning and the end of test (additive effect: 43.4 g/day) and fat depth measured in the mid-back position (additive effect: 1.82 mm). There was no evidence of interactions between the QTLs and sex, grandparents or F1 sires, suggesting that the detected QTLs were fixed for alternative alleles in the Meishan and Large White breeds. Comparison of locations suggests that these QTLs could be the same as those found in the Wild Boar x Large White cross.