Cold effect analysis and screening of SNPs associated with cold-tolerance in the olive flounder Paralichthys olivaceus

Olive flounder Paralichthys olivaceus is an important mariculture fish and cold stress limits to its growth and survival during winter. In the present study, the cold-tolerant (CT) and cold-sensitive (CS) flounder were defined after cold treatment (0.7 +/- 0.05 degrees C). Cold effects on histological and physiological and gene expression levels were analyzed. The correlation analyses between single nucleotide polymorphism locus (SNP) in hsp70, yb-1 and hmgb1 and the trait of cold tolerance were performed. The results showed that flounder gill was obviously damaged after treatment and more serious lesions in some of the epithelial cell layers were observed in the CS group. ATP concentration and pyruvate kinase activity in the CT group were significantly higher than those in the CS group (p < 0.05). Expression levels of hsp70 and yb-1 were both up-regulated markedly after cold treatment (p < 0.05). Nine, twenty-one and ten SNPs were screened from hsp70, yb-1 and hmgb1 partial sequences, respectively. Among them, 3 SNPs in hsp70 and 2 SNPs in hmgb1 showed significant difference in the two groups. Allele G of the SNP 8 (locus 1797) in hsp70 was only observed in the CT group. In the SNP 7 (locus 725) of hmgb1, genotype TT and allele T, and genotype CC and allele C were associated with cold tolerance and cold sensitive, respectively. Furthermore, one haplotype (TTG) generated from SNPs of hsp70 gene among CT and CS individuals showed significant relationship with cold tolerance (p = 0.031). Two haplotypes (ATG) and (TCT) generated from SNPs of hmgb1 were significantly associated with resistance (p = 0.001) and sensitive (p = 0.002) in cold treatment, respectively. These SNPs, genotype, alleles and haplotypes of hsp70 and hmgb1 may be related to the cold resistance of flounder, and could be candidate markers potentially for further selective breeding.