Polymorphism of Duck HSP70 Gene and mRNA Expression under Heat Stress Conditions
DOI:
https://doi.org/10.3923/ijps.2019.591.597Keywords:
Heat resistance, heat stress, HSP70, local ducks, mRNA expressions, polymorphismAbstract
Background and Objective: Tropical ambient temperature conditions that exceed comfortable temperatures for ducks and changes in maintenance systems from traditional to intensive conditions under low confined water cause ducks to have thermoregulation difficulties, causing the ducks to experience heat stress. The heat shock protein 70 (HSP70) gene, a known marker gene of heat resistance, can be a gene candidate for the selection of heat-resistant traits in local ducks to cope with heat stress. The objective of this study was to identify polymorphisms and the mRNA expression of the HSP70 gene under heat stress conditions. Materials and Methods: Genotyping was performed on 110 local duck breeds from West Sumatra (Pitalah, Bayang, Kamang and Payakumbuh). Polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) and Sanger sequencing assays were used to identify polymorphisms. Polymorphism analysis was conducted with the MEGA 7 software. mRNA expression was identified in 12 liver tissue samples from 12 local ducks (3 birds of each local duck breed) under control conditions or heat stress treatment performed at 35°C for 1 or 2 h. Results: The amplification product was 466-bp. HSP70/HhaI was polymorphic in all the breeds and two SNPs (Single nucleotide Polymorphism), g.1696G >A and g. 1762C >T, were recognized in all the HSP70/HhaI loci. The g.1696g >A locus produced two alleles (A and G) with three genotypes (AA, AG and GG) and the g. 1762C >T locus also produced two alleles (T and C) with three genotypes (TT, TC and CC). The analysis showed that the G allele had a higher frequency than that of the A allele and the T allele had a higher frequency than that of the allele C in all breeds. χ2 analysis showed that all the local duck breeds fit the Hardy-Weinberg equilibrium. The relative expression of the HSP70 gene mRNA in the liver and ovary tissues showed that heat stress had a significant effect (p<0.05) on the local ducks in the same breed. After 1 h of heat treatment, liver tissues from the Payakumbuh ducks showed the highest HSP70 mRNA expression, while after 2 h of treatment, the highest HSP70 mRNA expression was observed in the Bayang ducks. After the heat treatment of the ovarian tissues for 1 and 2 h, the Bayang ducks showed the highest HSP70 mRNA expression. Conclusion: HSP70 gene polymorphism and mRNA expression can be used as a candidate marker to increase heat resistance in ducks.
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