Chromosome localization of 19 β-hsd genes from orange-spotted grouper.

Phylogenetic analysis, gene structure and conserved domains of β-hsd genes in orange-spotted grouper. (A) The phylogenetic tree of all β-Hsd in grouper was constructed using Neighbor-Joining method. (B) The exon/intron organization of β-hsd genes of grouper. The red boxes represent exons and black lines indicate introns. (C) The conserved protein domains in the β-Hsd were identified using CD-Search program. Each domain is indicated with a specific color. Yellow: NADB_Rossmann superfamily, green: PLN02864 superfamily, purple: SCP2 superfamily, blue: SDR superfamily.

Phylogenetic relationships of β-Hsd proteins from grouper and other five species. The phylogenetic tree was constructed using MEGA7 by Maximum Likelihood method with the amino acid sequences of β-Hsd from grouper (Epinephelus coioides, Ec), (Homo sapiens, Hs), mouse (Mus musculus, Mm), zebrafish (Danio rerio, Dr), tilapia (Oreochromis niloticus, On), and stickleback (Gasterosteus aculeatus, Ga).

Clustering of the expression profiles of 19 grouper β-hsd genes during sex reversal. Genes were clustered according to phylogenetic relationships in expression profiles. The RPKM values were transformed into Z scores. Z scores were plotted according to Z = (x - μ)/σ, where x is the log 2 transformed gene expression measurement and μ and σ are the mean and standard deviations of expression of the gene. Red and blue colors indicate high and low relative expression levels after homogenization, respectively. The RPKM value for each gene and each stage was provided in the figure.

Quantitative RT-PCR examination of 10 β-hsd genes expressions in the brain and gonad during sex reversal of grouper. Values represent the relative mRNA expression in relation to internal control (β-actin gene). Data were expressed as the mean ± SEM for four replicates. In each panel, different lowercase letters indicate significant differences (p < 0.05).