DNA BARCODING OF FISH SPECIES FROM THE MEDITERRANEAN COAST OF ISRAEL

Accurately-classified genomic data in the Barcode of Life Data System (BOLD) database is vital to the protection and conservation of marine biodiversity in the Mediterranean Sea. The taxonomic classifications of 468 fish of 50 Mediterranean species were analyzed using the BOLD Identifier tool for variation in the cytochrome oxidase subunit I (COI) mitochondrial gene. Within species, nucleotide maximum composite likelihood was low with a mean of 0.0044±0.0008. Three presumptive species had significantly higher values e.g., Arnoglossus spp. (0.07), Torquigener flavimaculosus (0.013) and Boops boops (0.028). However, samples of Arnoglossus species were sub-classified into two groups that were finally identified as two different species e.g., Arnoglossus laterna and Arnoglossus thori. For the different species, BLAST searches against the BOLD database using our DNA barcoding data as the query sequences designated the most similar targets into groups. For each analyzed species, the similarity of the first and second threshold groups ranged from 95 to 99% and from 83 to 98%, respectively. Sequence based classification for the first threshold group was concordant with morphology-based identification. However, for 34 analyzed species (68%) overlaps of species between the two threshold groups hampered classification. Tree-based phylogeny analysis detected more than one cluster in the first threshold group for 22 out of 50 species, representing genetic subgroups and geographic origins. There was a tendency for higher conservation and lower number of clusters in the Lessepsian (Red Sea) migrant versus indigenous species

Linkage and Physical Mapping of Sex Region on LG23 of Nile Tilapia (Oreochromis niloticus)

Evidence supports that sex determination (SD) in tilapia is controlled by major genetic factors that may interact with minor genetic as well as environmental factors, thus implying that SD should be analyzed as a quantitative trait. Quantitative trait loci (QTL) for SD in Oreochromis niloticus were previously detected on linkage groups (LG) 1 and 23. Twenty-one short single repeats (SSR) of >12 TGs and one single nucleotide polymorphism were identified using the unpublished tilapia genome sequence on LG23. All markers showed two segregating alleles in a mapping family that was obtained by a cross between O. niloticus male (XY) and sex-reversed female (ΔXY) yielding 29 females (XX) and 61 males (XY and YY). Interval mapping analysis mapped the QTL peak between SSR markers ARO172 and ARO177 with a maximum F value of 78.7 (P < 7.6 × 10−14). Twelve adjacent markers found in this region were homozygous in females and either homozygous for the alternative allele or heterozygous in males. This segment was defined as the sex region (SR). The SR encompasses 1.5 Mbp on a single tilapia scaffold (no. 101) harboring 51 annotated genes. Among 10 candidate genes for SD that were tested for gene expression, anti-Müllerian hormone (Amh), which is located in the center of the SR, showed the highest overexpression in male vs. female embryos at 3 to 7 days postfertilization

Genomic insight into the common carp (Cyprinus carpio) genome by sequencing analysis of BAC-end sequences

<p>Abstract</p> <p>Background</p> <p>Common carp is one of the most important aquaculture teleost fish in the world. Common carp and other closely related Cyprinidae species provide over 30% aquaculture production in the world. However, common carp genomic resources are still relatively underdeveloped. BAC end sequences (BES) are important resources for genome research on BAC-anchored genetic marker development, linkage map and physical map integration, and whole genome sequence assembling and scaffolding.</p> <p>Result</p> <p>To develop such valuable resources in common carp (<it>Cyprinus carpio</it>), a total of 40,224 BAC clones were sequenced on both ends, generating 65,720 clean BES with an average read length of 647 bp after sequence processing, representing 42,522,168 bp or 2.5% of common carp genome. The first survey of common carp genome was conducted with various bioinformatics tools. The common carp genome contains over 17.3% of repetitive elements with GC content of 36.8% and 518 transposon ORFs. To identify and develop BAC-anchored microsatellite markers, a total of 13,581 microsatellites were detected from 10,355 BES. The coding region of 7,127 genes were recognized from 9,443 BES on 7,453 BACs, with 1,990 BACs have genes on both ends. To evaluate the similarity to the genome of closely related zebrafish, BES of common carp were aligned against zebrafish genome. A total of 39,335 BES of common carp have conserved homologs on zebrafish genome which demonstrated the high similarity between zebrafish and common carp genomes, indicating the feasibility of comparative mapping between zebrafish and common carp once we have physical map of common carp.</p> <p>Conclusion</p> <p>BAC end sequences are great resources for the first genome wide survey of common carp. The repetitive DNA was estimated to be approximate 28% of common carp genome, indicating the higher complexity of the genome. Comparative analysis had mapped around 40,000 BES to zebrafish genome and established over 3,100 microsyntenies, covering over 50% of the zebrafish genome. BES of common carp are tremendous tools for comparative mapping between the two closely related species, zebrafish and common carp, which should facilitate both structural and functional genome analysis in common carp.</p

B Chromosomes Have a Functional Effect on Female Sex Determination in Lake Victoria Cichlid Fishes

The endemic cichlid fishes in Lake Victoria are a model system for speciation through adaptive radiation. Although the evolution of the sex-determination system may also play a role in speciation, little is known about the sex-determination system of Lake Victoria cichlids. To understand the evolution of the sex-determination system in these fish, we performed cytogenetic analysis in 11 cichlid species from Lake Victoria. B chromosomes, which are present in addition to standard chromosomes, were found at a high prevalence rate (85%) in these cichlids. In one species, B chromosomes were female-specific. Cross-breeding using females with and without the B chromosomes demonstrated that the presence of the B chromosomes leads to a female-biased sex ratio in this species. Although B chromosomes were believed to be selfish genetic elements with little effect on phenotype and to lack protein-coding genes, the present study provides evidence that B chromosomes have a functional effect on female sex determination. FISH analysis using a BAC clone containing B chromosome DNA suggested that the B chromosomes are derived from sex chromosomes. Determination of the nucleotide sequences of this clone (104.5 kb) revealed the presence of several protein-coding genes in the B chromosome, suggesting that B chromosomes have the potential to contain functional genes. Because some sex chromosomes in amphibians and arthropods are thought to be derived from B chromosomes, the B chromosomes in Lake Victoria cichlids may represent an evolutionary transition toward the generation of sex chromosomes

Comparative physical maps derived from BAC end sequences of tilapia (Oreochromis niloticus)

Background: The Nile tilapia is the second most important fish in aquaculture. It is an excellent laboratory model, and is closely related to the African lake cichlids famous for their rapid rates of speciation. A suite of genomic resources has been developed for this species, including genetic maps and ESTs. Here we analyze BAC endsequences to develop comparative physical maps, and estimate the number of genome rearrangements, between tilapia and other model fish species. Results: We obtained sequence from one or both ends of 106,259 tilapia BACs. BLAST analysis against the genome assemblies of stickleback, medaka and pufferfish allowed identification of homologies for approximately 25,000 BACs for each species. We calculate that rearrangement breakpoints between tilapia and these species occur about every 3 Mb across the genome. Analysis of 35,000 clones previously assembled into contigs by restriction fingerprints allowed identification of longer-range syntenies. Conclusions: Our data suggest that chromosomal evolution in recent teleosts is dominated by alternate loss of gene duplicates, and by intra-chromosomal rearrangements (~one per million years). These physical maps are a useful resource for comparative positional cloning of traits in cichlid fishes. The paired BAC end sequences from these clones will be an important resource for scaffolding forthcoming shotgun sequence assemblies of the tilapia genome. (Résumé d'auteur

Microsatellite Support for Active Inbreeding in a Cichlid Fish

In wild animal populations, the degree of inbreeding differs between species and within species between populations. Because mating with kin often results in inbreeding depression, observed inbreeding is usually regarded to be caused by limited outbreeding opportunities due to demographic factors like small population size or population substructuring. However, theory predicts inclusive benefits from mating with kin, and thus part of the observed variation in inbreeding might be due to active inbreeding preferences. Although some recent studies indeed report kin mating preferences, the evidence is still highly ambiguous. Here, we investigate inbreeding in a natural population of the West African cichlid fish Pelvicachromis taeniatus which showed clear kin mating preferences in standardized laboratory experiments but no inbreeding depression. The presented microsatellite analysis reveals that the natural population has, in comparison to two reference populations, a reduced allelic diversity (A = 3) resulting in a low heterozygosity (Ho = 0.167) pointing to a highly inbred population. Furthermore, we found a significant heterozygote deficit not only at population (Fis = 0.116) but also at subpopulation level (Fis = 0.081) suggesting that inbreeding is not only a by-product of population substructuring but possibly a consequence of behavioral kin preferences

Hybrid Origin of the Thai-Chitralada Tilapia Strain Using DNA Barcoding and Microsatellite Analysis

The Thai-Chitralada strain originates from Egypt and was transferred to Japan. From there a stock of 50 fish were introduced to the Royal Chitralada Palace in Thailand. The commercially cultured strain of Nile tilapia O. niloticus in Thailand, Thai-Chitralada, possesses desirable traits for aquaculture, such as high growth rate integrated with reasonable fecundity and excellent performance in Asian countries. In 2010 a few dozen Thai-Chitralada fish were introduced to Israel from Thailand. The objective of this study is to trace the origin of Thai-Chitralada using DNA barcoding and microsatellite genetic markers. Cytochrome Oxidase I and D-loop sequences of 19 Thai-Chitralada fish clustered into three groups comprising 6, 7, and 6 individuals were homologous to the consensus sequences of O. aureus, O. niloticus, and O. mossambicus, respectively. The allele ranges for microsatellites UNH168 and G7A exclude O. niloticus (Ghana) as potential contributor to the ThaiChitralada strain. Genotyping for three microsatellites indicate overlap of alleles between Thai-Chitralada and O. niloticus (Egypt) and O. aureus, while O. mossambicus was not tested. Thus, our data based on mitochondrial and genomic analyses demonstrate that O. aureus, O. niloticus (Egypt), and O. mossambicus contributed to the formation of Thai-Chitralada. Thai-Chitralada can be used as a genetic resource for selection and adaptation to different geographical regions because of its diverse genetic background and desirable traits

Genetic Stock Identification of the Flathead Grey Mullet (Mugil cephalus) in Lake Tiberias Based on ParentOffspring Relationship

The Lake Tiberias (Sea of Galilee) grey mullet (Mugil cephalus) population originates from translocated wild-caught, or hatchery-reared, fish. The aim of the study was to identify the taxonomic status and stock origin of a sample of 32 mullet individuals caught in the Sea of Galilee, based on the mitochondrial Cytochrome Oxidase I (COI) sequence analysis and nuclear microsatellite markers. A total of 13 microsatellite markers were selected from nine different linkage groups with a number of alleles ranging from 5-23, with an average of 9.85 alleles/locus. By using COI sequences and the Barcode of Life Data (BOLD) identification system, seven individuals were taxonomically classified as Thinlip grey mullet (Liza ramda) and the remaining 25 as M. cephalus. Of 663 nucleotides, 122 (18.4%) differed between the COI sequences of the two distinct mullet species. A preliminary parentage analysis of the hatchery-reared stocking batch, based on 13 markers, assigned them as progeny of three mating pairs. These three mating pairs only were tested as potential parents of the 25 M. cephalus individuals captured from the lake. Marker-based comparison showed that a parent-offspring relationship was rejected for 17 M. cephalus individuals by at least four genetic markers. Eight of 25 M. cephalus individuals (32%), were identified as progeny of the three parental pairs with the overall probability of 3.88 x 10-7 for correspondence by chance to any of these three parental pairs. The 13 markers used has high statistical power to reject a putative parent-offspring relationship obtained by chance thus resulting in familial identification. This approach represents an accurate method of genetic stock identification and should also be applicable to populations of other species. The results confirm that the mullet hatcheryreared fingerlings survived in the Sea of Galilee when released alongside wildcaught fingerlings