Applications of microsatellite markers to genetic management of carps in aquaculture

Carp aquaculture in South Asia suffers severely from a lack of genetic management, which has eroded the genetic quality of both captive and wild populations. Use of molecular markers, especially microsatellites, has revolutionized genetic management of hatchery stocks through its ability to detect kinship between individuals and hence in controlling level of inbreeding and loss of genetic diversity. In the present PhD work, microsatellite markers were applied to breeding programmes for silver carp (Hypophthalmichthys molitrix) and common carp (Cyprinus carpio) to study different genetic management aspects and new markers were generated from rohu (Labeo rohita). A set of newly isolated microsatellite markers from silver carp were characterized and two pentaplex PCR reactions were optimized to enable rapid genotyping of large number of individuals at 10 microsatellite loci. The utility of these markers in parentage, sibship and relatedness analysis were assessed by applying them to groups of fish with known relationship. These markers were used for parentage analysis in a breeding programme designed to estimate heritability of harvest weight and length in silver carp. Full- and half-sib families were created in three sets of partly factorial mating and all the families from each set were reared in communal ponds from very early life stages. With ten microsatellites 96.3% of the offspring could be assigned to a single family. Heritability estimates were found to be 0.65 ± 0.13 for weight and 0.50 ± 0.13 for length. High estimates of h2 suggested that this population should respond rapidly to selection for increased harvest size. Microsatellite markers were also applied to monitor the early stages of a mass selection programme in common carp (Cyprinus carpio). The selection was initiated from a base population synthesized from six different stocks. The selected individuals were divided to create two separate lines. The aims of this study were to monitor whether the stocks were represented in the intended proportions in the F1 selected populations, to investigate the relative contribution of families and its impact on effective population size and to identify any loss of molecular genetic variation. Five highly polymorphic microsatellites were used for parentage analysis of the selected fish to track stock and family contribution. Overall, large perturbations were observed in the relative contributions of two major stocks. Family contribution was also highly variable, causing the Ne to drop to below half the census size. A loss of 6.9%-12.2% of microsatellite alleles was observed but loss of heterozygosity was not very prominent. The replicate lines showed significant differences in allelic distribution after the first generation of selection, but not in genotypic distribution. Finally, 52 microsatellite markers were isolated from a partial genomic library of rohu using a selective hybridization protocol. Characterization of these markers resulted in 36 polymorphic loci, which will be useful in future work on conservation and management of both wild and captive rohu populations

Genome-wide characterization of genetic variants and putative regions under selection in meat and egg-type chicken lines

Abstract\ud \ud Background\ud Meat and egg-type chickens have been selected for several generations for different traits. Artificial and natural selection for different phenotypes can change frequency of genetic variants, leaving particular genomic footprints throghtout the genome. Thus, the aims of this study were to sequence 28 chickens from two Brazilian lines (meat and white egg-type) and use this information to characterize genome-wide genetic variations, identify putative regions under selection using Fst method, and find putative pathways under selection.\ud \ud \ud Results\ud A total of 13.93 million SNPs and 1.36 million INDELs were identified, with more variants detected from the broiler (meat-type) line. Although most were located in non-coding regions, we identified 7255 intolerant non-synonymous SNPs, 512 stopgain/loss SNPs, 1381 frameshift and 1094 non-frameshift INDELs that may alter protein functions. Genes harboring intolerant non-synonymous SNPs affected metabolic pathways related mainly to reproduction and endocrine systems in the white-egg layer line, and lipid metabolism and metabolic diseases in the broiler line. Fst analysis in sliding windows, using SNPs and INDELs separately, identified over 300 putative regions of selection overlapping with more than 250 genes. For the first time in chicken, INDEL variants were considered for selection signature analysis, showing high level of correlation in results between SNP and INDEL data. The putative regions of selection signatures revealed interesting candidate genes and pathways related to important phenotypic traits in chicken, such as lipid metabolism, growth, reproduction, and cardiac development.\ud \ud \ud Conclusions\ud In this study, Fst method was applied to identify high confidence putative regions under selection, providing novel insights into selection footprints that can help elucidate the functional mechanisms underlying different phenotypic traits relevant to meat and egg-type chicken lines. In addition, we generated a large catalog of line-specific and common genetic variants from a Brazilian broiler and a white egg layer line that can be used for genomic studies involving association analysis with phenotypes of economic interest to the poultry industry.CB received a fellowship from the program Science Without Borders - National Council for Scientific and Technological Development (CNPq, grant 370620/2013–5). GCMM and TFG received fellowships from São Paulo Research Foundation (FAPESP, grants 14/21380–9 and 15/00616–7). LLC is recipient of productivity fellowship from CNPq. This project was funded by São Paulo Research Foundation (FAPESP) - thematic project (2014/08704–0)