Complete mitochondrial genome of the hybrid grouper Epinephelus lanceolatus (♀) × E. moara (♂).

The complete mitochondrial genome of the novel hybrid grouper (Epinephelus lanceolatus ♀ and E. moara ♂) includes 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNA genes, and 1 control region (D-loop), for a total length of 16,743 bp. The overall nucleotide compositions encoded on the heavy strand are 29.73% A, 28.84% C, 15.08% G, and 26.64% T.

The complete mitochondrial genome of the hybrid grouper Epinephelus lanceolatus (♀) × E. akaara (♂).

This study determined the complete mitochondrial genome of the hybrid grouper Epinephelus lanceolatus ♀ × E. akaara ♂. The complete mitochondrial genome is 16,743 bp long and includes 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and a control region. The nucleotide composition of the L-strand was as follows: A 20.00, C 14.10, G 29.21, and T 36.68%. All of the genes are encoded on the H-strand, except for NADH dehydrogenase subunit (ND6) and eight tRNA genes. A phylogenetic tree constructed using the maximum-likelihood (ML) method showed that the hybrid grouper E. lanceolatus ♀ × E. akaara ♂ is similar to E. lanceolatus.

Development of novel tetra- and trinucleotide microsatellite markers for giant grouper Epinephelus lanceolatus using 454 pyrosequencing.

Giant grouper (Epinephelus lanceolatus) is a commercially important species, but its wild population has recently been classified as vulnerable. This species has significant potential for use in aquaculture, though a greater understanding of population genetics is necessary for selective breeding programs to minimize kinship for genetically healthy individuals. High-throughput pyrosequencing of genomic DNA was used to identify and characterize novel tetra- and trinucleotide microsatellite markers in giant grouper from Sabah, Malaysia. In total, of 62,763 sequences containing simple sequence repeats (SSRs) were obtained, and 78 SSR loci were selected to possibly contain tetra- and trinucleotide repeats. Of these loci, 16 had tetra- and 8 had trinucleotide repeats, all of which exhibited polymorphisms within easily genotyped regions. A total of 143 alleles were identified with an average of 5.94 alleles per locus, with mean observed and expected heterozygosities of 0.648 and 0.620, respectively. Among of them, 15 microsatellite markers were identified without null alleles and with Hardy-Weinberg equilibrium. These alleles showed a combined non-exclusion probability of 0.01138. The probability of individual identification (PID) value combined with in descending order 12 microsatellite markers was 0.00008, which strongly suggests that the use of the microsatellite markers developed in this study in various combinations would result in a high resolution method for parentage analysis and individual identification. These markers could be used to establish a broodstock management program for giant grouper and to provide a foundation for genetic studies such as population structure, parentage analysis, and kinship selection.

Genomic organization and functional diversification of two warm-temperature-acclimation-associated 65-kDa protein genes in rockbream (Oplegnathus fasciatus; Perciformes).

Two paralogue genes of warm-temperature-acclimation-associated 65-kDa protein were characterized and their mRNA expression patterns during various experimental stimulations were examined in the rockbream (Oplegnathus fasciatus; Perciformes). Rockbream Wap65 isoforms (rbWap65-1 and rbWap65-2) share basically common structural features with other teleostean orthologues and human hemopexin (HPX) at both amino acid (conserved cysteine and histidine residues) and genomic levels (ten-exon structure), although the rbWap65-2 reveals more homologous characteristics to human HPX than does rbWap65-1 isoform. Southern blot analysis indicates that each rbWap65 isoform exists as a single copy gene in the rockbream genome. Both rbWap65 genes were predicted to possess various transcription factor (TF) binding motifs related with stress and innate immunity in their 5ʹ-upstream regions, in which inflammation-related motifs were more highlighted in the rbWap65-2 than in rbWap65-1. Based on the RT-PCR assay, the liver-predominant expression pattern was more apparent in rbWap65-1 than rbWap65-2 isoform. During thermal elevation, clear upregulation was found only for the rbWap65-1. In contrast, immune stimulations (bacterial challenges, viral infection and iron overload) activated more preferentially the rbWap65-2 isoform in overall, although the inducibility was affected by the kinds of stimulators and tissue types. Taken together, our data suggest that the two paralogue rbWap65 isoforms have experienced subfunctionalization and/or neofunctionalization during their evolutionary history, in which the rbWap65-2 has retained closer, functional orthology to the human HPX while the rbWap65-1 have been diversified to be more related with thermal acclimation physiology.

Induction of Primary Male in Juvenile Red Spotted Grouper Epinephelus akaara by Immersion of 17α-Methyltestosterone.

We investigated the androgenic effects of 17α- methyltestosterone (MT) on gonadal sex reversal in juvenile red spotted grouper Epinephelus akaara. The fish were immersed in 17α-MT at 1 and 5 mg/L. Treatment method of 17α-MT was once weekly for 4 and 8 weeks. Fish were sampled at 12 months after end of the treatment period in order to histological analysis. At the initiation of an experiment (70 day after hatching), juvenile red spotted grouper have the paired primordial gonads with somatic cells bellow kidney in the posterior portion of the body cavity. Formation of ovarian cavity indicates that the ovarian differentiation beginning at 70 DAH in red spotted grouper. At 12 months after end of the treatment period, control group, 17α-MT 1 mg/L treatment group for 4 and 8 weeks, and 17α-MT 5 mg/L treatment group for 4 weeks were all female. However, sex-changed males without ovarian cavity were observed in the 17α-MT 5 mg/L treatment group for 8 weeks. In grouper, we firstly reported that the red spotted grouper be able to induce the primary males by hormone treatment prior to gonadal sex differentiation.

Induced Morphological Changes in Larval Rock Bream, Oplegnathus fasciatus, under Starvation.

Morphological changes in the reared rock bream, Oplegnathus fasciatus, from hatching to six days after hatching were examined during the early growth stage under starvation. All the larvae died within five days when feeding was delayed for three days after hatching. These results imply that initial larval food should be supplied within two days of hatching. Changes in the pectoral angle and the ratios of eye height to head height, gut height to standard length, and gut height to myotome height in the rock bream are alternative indicators for the identification of starving fish. These indicators might prove useful in evaluating the successful transition from endogenous to exogenous feeding in this species.