Genetic variability, structure and assignment of Spanish and French pig populations based on a large sampling.

The Spanish and French pig populations share the common practice of quasi systematic paternity control of pure breed and composite line males. Ten microsatellite markers are in common between Spain and France controls, among the 17 markers used in France and the 13 used in Spain. After the adjustment of allele sizes, it is possible to merge the two datasets and to obtain a set of 5791 animals, including the vast majority of the males in the Duroc, Landrace, Large White and Piétrain French and Spanish breeds. Twelve French composite lines are also available. The genetic diversity analysis of these pig populations is presented, as well as the assignment of an individual to its breed. The effects of heterogeneous sampling across time and of relatedness among animals are also assessed. Consistent with the results of the previous studies, we found that different populations from the same breed clearly clustered together. In addition, all populations of this study, whether purebred or composite, are quite well differentiated from the other ones. As a result, we note that the 10 microsatellites commonly used for paternity control ensure a powerful detection of the breed of origin, with the power of detection being 95-99%. The detection of the exact population within breed is more difficult, but the power exceeds 70% for most of the populations. Practical implications include, for instance, the detection of outlier animals, crosses and admixture events.

A set of 37 microsatellite DNA markers for genetic diversity and structure analysis of Atlantic salmon Salmo salar populations.

Atlantic salmon Salmo salar microsatellite markers from a large database were analysed and selected with technical, economic and genetic criteria to provide an optimized set of polymorphic DNA markers for the analysis of the genetic diversity and the structure of anadromous Atlantic salmon populations. A set of 37 microsatellite markers was identified that are easy to use and provide a high level of differentiation power.

Using markers in gene introgression breeding programs.

We investigate the use of markers to hasten the recovery of the recipient genome during an introgression breeding program. The effects of time and intensity of selection, population size, number and position of selected markers are studied for chromosomes either carrying or not carrying the introgressed gene. We show that marker assisted selection may lead to a gain in time of about two generations, an efficiency below previous theoretical predictions. Markers are most useful when their map position is known. In the early generations, it is shown that increasing the number of markers over three per non-carrier chromosome is not efficient, that the segment surrounding the introgressed gene is better controlled by rather distant markers unless high selection intensity can be applied, and that selection on this segment first can reduce the selection intensity available for selection on non-carrier chromosomes. These results are used to propose an optimal strategy for selection on the whole genome, making the most of available material and conditions (e.g., population size and fertility, genetic map).

Identity coefficients in finite populations. I. Evolution of identity coefficients in a random mating diploid dioecious population.

Properties of identity relation between genes are discussed, and a derivation of recurrent equations of identity coefficients in a random mating, diploid dioecious population is presented. Computations are run by repeated matrix multiplication. Results show that for effective population size (Ne) larger than 16 and no mutation, a given identity coefficient at any time t can be expressed approximately as a function of (1--f), (1--f)3 and (1--f)6, where f is the mean inbreeding coefficient at time t. Tables are presented, for small Ne values and extreme sex ratios, showing the pattern of change in the identity coefficients over time. The pattern of evolution of identity coefficients is also presented and discussed with respect to Neu, where u is the mutation rate. Applications of these results to the evolution of genetic variability within and between inbred lines are discussed.

Detection of quantitative trait loci for fat androstenone levels in pigs.

A QTL analysis of fat androstenone levels from a three-generation experimental cross between Large White and Meishan pig breeds was carried out. A total of 485 F2 males grouped in 24 full-sib families, their 29 parents and 12 grandparents were typed for 137 markers distributed over the entire porcine genome. The F2 male population was measured for fat androstenone levels at 100, 120, 140, and 160 d of age and at slaughter around 80 kg liveweight. Statistical analyses were performed using two interval mapping methods: a line-cross (LC) regression method, which assumes alternative alleles are fixed in founder lines, and a half- full-sib (HFS) maximum likelihood method, where allele substitution effects were estimated within each half- and full-sib family. Both methods revealed genomewide significant gene effects on chromosomes 3, 7, and 14. The QTL explained, respectively, 7 to 11%, 11 to 15%, and 6 to 8% of phenotypic variance. Three additional significant QTL explaining 4 to 7% of variance were detected on chromosomes 4 and 9 using LC method and on chromosome 6 using HFS method. Suggestive QTL were also obtained on chromosomes 2, 10, 11, 13, and 18. Meishan alleles were associated with higher androstenone levels, except on chromosomes 7, 10, and 13, although 10 and 13 additive effects were near zero. The QTL had essentially additive effects, except on chromosomes 4, 10, and 13. No evidence of linked QTL or imprinting effects on androstenone concentration could be found across the entire porcine genome. The steroid chromosome P450 21-hydroxylase (CYP21) and cytochrome P450 cholesterol side chain cleavage subfamily XIA (CYP11A) loci were investigated as possible candidate genes for the chromosome 7 QTL. No mutation of coding sequence has been found for CYP21. Involvement of a candidate regulatory mutation of CYP11A gene proposed by others can be excluded in our animals.

Genetic linkage mapping of quantitative trait loci for behavioral and neuroendocrine stress response traits in pigs.

A QTL analysis of behavioral and neuroendocrine responses to a "novel environment" stress was conducted in a three-generation experimental cross between Meishan and Large White pig breeds. A total of 186 F2 males and 182 F2 females were studied for their behavioral and neuroendocrine reactivity to a novel environment test at 6 wk of age. Locomotion, vocalization, and defecation rate, as well as exploration time, were measured for 10 min. Blood samples were taken immediately before and after the test to measure plasma levels of ACTH, cortisol, and glucose. Animals were typed for a total of 137 markers covering the entire porcine genome. Analyses were performed using two interval mapping methods: a line-cross regression method, where founder lines were assumed to be fixed for different QTL alleles, and a half-/full-sib maximum likelihood method where allele substitution effects were estimated within each half-/full-sib family. Both methods revealed a highly significant gene effect for poststress cortisol level (P < 0.001) and a significant effect for basal cortisol level (P < 0.05) at the end of the q arm of chromosome 7, explaining, respectively, 20% and 7% of the phenotypic variance. Meishan alleles are associated with higher cortisol levels and are partially dominant (for poststress levels) over Large White alleles. Other significant gene effects on biological measures were detected on chromosomes 1 and 17 (ACTH response to stress), 3, 5, and 8 (glucose levels). The SSC 17 QTL explains 12% of the phenotypic variance of poststress ACTH levels, with a suggestive evidence of imprinting effects. Meishan alleles are associated with lower poststress ACTH levels. Gene effects of low amplitude only were found for behavioral reactivity traits. Considering the effects of stress neuroendocrine systems on energy fluxes and protein deposition, and the importance of stress reactivity for meat quality and animal welfare, these results open new perspectives for pig selection.

Genetic diversity within and between European pig breeds using microsatellite markers.

An important prerequisite for a conservation programme is a comprehensive description of genetic diversity. The aim of this study was to use anonymous genetic markers to assess the between- and the within-population components of genetic diversity for European pig breeds at the scale of the whole continent using microsatellites. Fifty-eight European pig breeds and lines were analysed including local breeds, national varieties of international breeds and commercial lines. A sample of the Chinese Meishan breed was also included. Eleven additional breeds from a previous project were added for some analyses. Approximately 50 individuals per breed were genotyped for a maximum of 50 microsatellite loci. Substantial within-breed variability was observed, with the average expected heterozygosity and observed number of alleles per locus being 0.56 [range 0.43-0.68] and 4.5 respectively. Genotypic frequencies departed from Hardy-Weinberg expectations (P < 0.01) in 15 European populations, with an excess of homozygotes in 12 of them. The European breeds were on average genetically very distinct, with a Wright F(ST) index value of 0.21. The Neighbour-Joining tree drawn from the Reynolds distances among the breeds showed that the national varieties of major breeds and the commercial lines were mostly clustered around their breeds of reference (Duroc, Hampshire, Landrace, Large White and Piétrain). In contrast, local breeds, with the exception of the Iberian breeds, exhibited a star-like topology. The results are discussed in the light of various forces, which may have driven the recent evolution of European pig breeds. This study has consequences for the interpretation of biodiversity results and will be of importance for future conservation programmes.

Genetic diversity in European pigs utilizing amplified fragment length polymorphism markers.

The use of DNA markers to evaluate genetic diversity is an important component of the management of animal genetic resources. The Food and Agriculture Organisation of the United Nations (FAO) has published a list of recommended microsatellite markers for such studies; however, other markers are potential alternatives. This paper describes results obtained with a set of amplified fragment length polymorphism (AFLP) markers as part of a genetic diversity study of European pig breeds that also utilized microsatellite markers. Data from 148 AFLP markers genotyped across samples from 58 European and one Chinese breed were analysed. The results were compared with previous analyses of data from 50 microsatellite markers genotyped on the same animals. The AFLP markers had an average within-breed heterozygosity of 0.124 but there was wide variation, with individual markers being monomorphic in 3-98% of the populations. The biallelic and dominant nature of AFLP markers creates a challenge for their use in genetic diversity studies as each individual marker contains limited information and AFLPs only provide indirect estimates of the allelic frequencies that are needed to estimate genetic distances. Nonetheless, AFLP marker-based characterization of genetic distances was consistent with expectations based on breed and regional distributions and produced a similar pattern to that obtained with microsatellites. Thus, data from AFLP markers can be combined with microsatellite data for measuring genetic diversity.

The number of lines of descent and fixation probabilities of alleles in the pure genetic drift process: analytical approximations.

Analysis of diversity between populations diverged from an evolutionarily small number of generations cannot be done under the assumption that allele frequencies reflect an equilibrium between genetic drift and mutations. An alternative to analysis through coalescence theory is proposed in this situation by developing analytical approximations. Using a Poisson approximation of its distribution, the expected number of genes from one generation whose copies make up the population after a number of generations characterized by fixation index F can be shown to be approximated by 2/F-1, irrespective of population size, and probabilities of fixation of alleles over a finite period of time can be also approximated. These expressions, which were checked numerically, should make it possible to calculate approximate likelihoods for allele frequency distributions promoted by drift.

Statistical decision rules concerning synteny or independence between markers.

Data on the segregation of human markers in somatic hybrids between permanent rodent cell lines and primary human cells were gathered and statistically analyzed, using various criteria of association. The analysis provides evidence that human chromosomes do not segregate independently in somatic cell hybrids. A statistical decision rule concerning synteny or independence between markers is proposed, and its utility in developing the gene maps of other species by means of somatic cell hybridization is explored.

An algorithm for comparing RNA secondary structures and searching for similar substructures.

To access the functional informations carried by RNA molecules at the level of their secondary structure interactions, we propose a comparison method based on a tree edit algorithm which takes into account the tree structure of RNA foldings. Any secondary structure is translated into a tree involving all its elementary substructures; then a shorter condensed tree is built in which any unbranched helix interspersed with bulges and interior loops is taken as a single node. This method includes several parameters: a comparison matrix between structural units, gap penalties, and the scoring between nodes of the condensed trees. Their effects have been analysed using as a model a rapidly divergent domain of the large ribosomal RNA, for which structural variation during evolution is well known. This method allows one to recognize precisely, in large target molecules, definite substructures that present with the query molecules only a limited set of closely related secondary structure features; it is still efficient if intervening features, which can correspond to insertion/deletion of entire stem regions, separate such structural elements. When coupled with a hierarchical clustering algorithm, this method is suitable for classifying RNA molecules according to their secondary structure homologies.

Interactions of selection, linkage and drift in the dynamics of polygenic characters.

We study the dynamics under directional truncation selection of the genetic variability of a quantitative character controlled by a finite number of possibly linked loci with additive effects. After the first generation of selection, the build-up of linkage disequilibria (Bulmer effect) is analytically demonstrated from a genetical point of view in an infinite population. In the following generations, the dynamics of the system in a finite population are predicted using analytic recurrences under a multi-normal approximation, and computer simulations. The effects of recombination on the dynamics of linkage disequilibria induced by selection and drift, and the consequences for the additive genetic variance are then analysed and discussed from the simulation results. Compared to the rapid exploitation of genetic variability promoted by high recombination rates, low recombination rates promote an early storage of genetic variability in repulsion associations of alleles and a possible late release of genetic variance in the population, so that the variability of the character may be maintained over a longer period of time. In some cases, favourable recombination events in tightly linked systems induce an increase of the additive variance of the character, which may explain some results observed in long-term selection experiments. Our results emphasize that the joint effects of selection, linkage and drift must not be neglected in theoretical quantitative genetics, and require further investigation.

Effects of population size and linkage on optimal selection intensity.

Following Robertson (1970a) it is generally considered that for mass selection the selected proportion that maximises ultimate response is 0.5. This prediction has been partly tested by different authors. Here we explicitly address the question using computer simulations of selection in finite populations with linkage. The results show that the response achieved is always lower than the one predicted by standard methods, and that optimum selection intensity may be much lower than predicted unless population size is small.

[A model of enzymatic kinetics]. / Sur un modèle en cinétique enzymatique.

In this Note, we study a system of differential equations representing the kinetics of an enzymatic reaction. For a closed system, and in the domain where it has a biological significance, it is shown that: (i) there exists a unique equilibrium point, which is an asymptotically stable point; (ii) if several enzymes act on the same substrates, the equilibrium values of the substrates concentrations take their values within the range of the equilibrium values achieved with each of these enzymes, (iii) the kinetics of the reaction can be approached by a single first-order differential equation, which may be seen as a generalization of the Michaelis equation.

Genetic pollution and number of matings in a black honey bee (Apis mellifera mellifera) population.

In the south-east of France, local honey bees possess only the B allele at the MDH locus, whereas the races which are usually imported into this area do not have this allele. The proportion of non-B genes in a sample of drones was used to measure the "genetic pollution" in the local population. Within the course of a breeding scheme of local bees, 99 queens, whose genotypes are BB, were naturally mated between April 25 and June 10, 1985 at la Tave (Gard, France). Twenty daughters-workers of each queen were analysed at the MDH locus. The frequency of the B allele in drones that mated with these queens is estimated by the proportion of workers with genotype BB and the genetic pollution by the cumulated frequency of the other alleles. The sampling variances of these frequencies involve a coefficient which is a function of the average number of drones mated with a queen. This latter parameter is estimated through the maximum likelihood method. In addition to the three well-known alleles, a rare allele (frequency=0.0055), possibly equivalent to the S1 allele described by Badino et al. (1983), has been found in three different colonies. Cumulating the frequencies of the non-B alleles results in an estimation of the genetic pollution equal to 0.0394 (±0.0071). This low value allows us to proceed to the next step of the selection project. The mean number of drones mated to a queen is 12.4 with a (10.4-19.3) confidence interval at the 90% level.

Optimal structure of protocol designs for building genetic linkage maps in livestock.

We investigated protocol designs for gene mapping in livestock. The optimization of the population structure was based on the empirical variance of the recombination rate estimator. We concluded that a mixture of half-sib and full-sib families is preferred to half-sib families; a knowledge of parental phases does not improve the quality of the estimation for typical livestock families with five offspring or more; and measurements of the genotype of the mates in half-sib families are not useful. Graphs and algebraic approximations for the practical choice of family size and structure are given.

Regional assignment of genetic markers using a somatic cell hybrid panel: a WWW interactive program available for the pig genome.

MOTIVATION: Quick and easy gene mapping by the use of a panel of cytogenetically characterized somatic cell hybrids is possible, even if some discordant experimental results arise. RESULTS: An interactive program is proposed and is made available on a WWW site to users of a somatic cell hybrid panel. Assignments to chromosomes and subchromosomal regions are based on likelihood calculations and Bayes' theorem, and a confidence level is provided. The method is illustrated in the case of the pig genome.

Localization of pig Na+,K(+)-ATPase alpha and beta subunit genes to chromosome 4 by radioactive in situ hybridization.

Two genes coding for Na+,K(+)-ATPase alpha and beta subunits are localized on pig chromosome 4, to the q1.6-->q2.3 and 1.3-->q2.1 regions, respectively, by radioactive in situ hybridization. According to nucleotide and amino acid sequence comparisons with different human isoforms of Na+,K(+)-ATPase, these pig alpha beta ATPase genes show strong homologies with human alpha 1 and beta subunit ATPase genes, respectively. These results are discussed with respect to comparative mapping data of conserved genes in mammalian species. We showed that the pig cDNA probes encoding ATPase alpha and beta genes reveal DNA polymorphism in Meishan and Large White pigs.

Genetic analysis of fingerprints in Mérinos d'Arles x Booroola Merino crossbred sheep.

The M13.13 minisatellite probe, consisting of a polymer of the M13 VNTR consensus sequence, cross-hybridized to ovine DNA and allowed detection of several polymorphic loci. Individual specific patterns were obtained in sheep using this probe. Pedigree analysis showed that individuals were heterozygous for most of the DNA fragments detected (88%). By studying the segregation of male's variable DNA fragments, a minimum of 10 loci were defined. The ovine DNA 'fingerprint' obtained with M13.13 is polymorphic enough to be used efficiently in animal identification, paternity testing, and possibly as a source of genetic markers for linkage analysis.

Genetic components of litter size variability in sheep.

Classical selection for increasing prolificacy in sheep leads to a concomitant increase in its variability, even though the objective of the breeder is to maximise the frequency of an intermediate litter size rather than the frequency of high litter sizes. For instance, in the Lacaune sheep breed raised in semi-intensive conditions, ewes lambing twins represent the economic optimum. Data for this breed, obtained from the national recording scheme, were analysed. Variance components were estimated in an infinitesimal model involving genes controlling the mean level as well as its environmental variability. Large heritability was found for the mean prolificacy, but a high potential for increasing the percentage of twins at lambing while reducing the environmental variability of prolificacy is also suspected. Quantification of the response to such a canalising selection was achieved.