ABSTRACT. This study aimed to estimate changes in HSF3 and HSP70 mRNA expression in stress-sensitive tissues of 2 chicken breeds during acute heat stress. Lingshan chickens (LSC) and White Recessive Rock (WRR) (24 chickens of each breed) were randomly divided into 4 groups (0, 2, 3, and 6 h of heat treatment). With increasing heat treatment time, both HSF3 and HSP70 expression first declined and then showed a significant increase in both breeds. However, HSP70 expression decreased in the heart following 6 h of heat treatment, whereas HSF3 expression continued to increase. After 2 h of heat treatment, HSF3 expression was significantly higher in the brain and leg muscle of LSC compared to WRR (P < 0.05, P < 0.01). In comparison, HSP70 expression was significantly higher in the liver and leg muscle of WRR compared to LSC (P < 0.01, P < 0.05). After 3 h of heat treatment, HSF3 expression was significantly higher in the brain and leg muscle of LSC compared to WRR (P < 0.01). In comparison, HSP70 expression was significantly higher in the liver and heart of LSC compared to WRR (P < 0.01). These results indicate that the expression of HSF3 and HSP70 mRNA in LSC and WRR exhibit species-specific and tissue-specific differences during heat treatment.

ABSTRACT. The Anyi tile-like gray chicken is a Chinese indigenous breed with a gray dilution phenotype, having gray feathers, comb, skin, shanks, and beak, which is valuable for genetic research on pigmentation. However, the genetic basis of the gray dilution phenotype remains unknown. The objective of this study was to investigate the genetic basis of the gray dilution phenotype in the Anyi tile-like gray chicken. We found that all Anyi tile-like gray chickens tested in this study carried at least one E allele, which is responsible for the appearance of black feathers, and some of them carried the FM allele, which is responsible for the black skin phenotype. A single nucleotide polymorphism (C.1909A>G) was identified within the melanophilin (MLPH) gene and was significantly associated with the gray dilution phenotype. Our findings suggest that the E and FM alleles act together to cause the development of the "five-black" phenotype (black feather, comb, skin, shank, and beak), whereas the MLPH mutation results in defective melanosome transport, leading to the development of the "five-gray" phenotype.

ABSTRACT. This study was designed to detect the sequence variation of the chicken heat shock protein 70 (HSP70) gene. A total of 102 individuals from 8 native Chinese breeds together with Dwarf White Chicken and Red Junglefowl were used to detect sequence variations. The coding regions of the chicken HSP70 gene from 102 individuals were cloned and sequenced. Thirty-six variations were identified, which included 34 single nucleotide polymorphisms and 2 indel mutations. Fifty-seven haplotypes were observed, of which, 43 were breed-specific and 14 were shared. There were 7 Red Junglefowl-specific haplotypes, while Haidong and Silkie only had 2 specific haplotypes. Eleven and 3 haplotypes were shared between and within species, respectively. The variation in nucleotide diversity (Pi) and average number of nucleotide (2015) differences (K) among species were consistent. The total Pi of HSP70 was 0.0016, and the total K was 4.1998. The Pi value of Red Junglefowl was the highest (0.0018) and K was 4.8000, while the Pi of Silkie was the lowest (0.0010) and K was 2.5000. These results demonstrated that variation in chicken HSP70 was abundant between and within species.

Somatotropic axis-related genes contribute to reproduction of ducks. Five SNP in the 5'-flanking regions of the growth hormone (GH), prolactin (PRL), and pituitary-specific transcription factor (Pit-1) genes were identified and genotyped in a female population of Muscovy ducks. Association analysis of these SNP with Muscovy duck egg production traits was performed. Results showed that SNP C-515G of GH was significantly associated with egg number in ducks at age 59 wk (E59W; P = 0.0009) and egg number in ducks at age 300 d (E300D; P = 0.0022). Single nucleotide polymorphism C-441T of GH was significantly associated with E59W (P = 0.0014). Significant associations of SNP T-884C and T-335C of PRL with the age at first egg (A1D), E59W, and E300D were detected in this population (P < 0.0001). It was concluded that these 4 SNP might be useful markers to use with the aim of increasing Muscovy duck E59W. On the basis of genetic parameter estimation, the heritability of A1D, E300D, E59W, and molting time were 0.43 ± 0.04, 0.45 ± 0.04, 0.36 ± 0.04, and 0.04 ± 0.03, respectively. Strong positive genetic correlation was noted between E59W and E300D (correlation coefficient = 0.80), whereas a negative association was noted between E59W and A1D (correlation coefficient = -0.80). Therefore, the selection for improved A1D should also increase E59W.

ABSTRACT. Studying thermotolerance is important for the prevention of thermostress in chickens. This study aimed to analyze the effect of mutations in the heat shock protein 70 (HSP70) gene on chicken thermotolerance. The C.-69A>G SNP in the 5'-flanking region of the HSP70 gene was genotyped in Lingshan and White Recessive Rock (WRR) chickens. Association of this SNP with thermotolerance traits revealed it to be significantly associated with CD4+/CD8+, and potentially associated with heterophil-to-lymphocyte ratio in WRR chickens exposed to thermoneutral temperature (15°C). Online prediction detected a putative myeloid zinc finger protein 1 binding factor in the C.-69A>G mutation. Under acute thermostress, mRNA levels of HSP70 in individuals with different C.-69A>G genotypes varied in the heart, leg muscle, and liver tissues. The HSP70 protein was expressed at higher levels in individuals with the GG genotype than in those with the AA genotype. In heart and liver, protein expression of HSP70 in individuals with the GG genotype was significantly higher than in those with the AA genotype. In leg muscle, protein expression was higher in birds with the GG genotype than in those with the AA and AG genotypes. Luciferase activity of the GG genotype was significantly higher than that of the AA genotype, suggesting that the C.-69A>G SNP regulates HSP70 gene expression. These results indicate that the C.-69A>G SNP in the 5'-flanking region of the HSP70 gene might affect chicken thermotolerance and that the GG genotype might be advantageous for the prevention of thermostress.