A Novel in Duck Myoblasts: The Transcription Factor Retinoid X Receptor Alpha (RXRA) Inhibits Lipid Accumulation by Promoting CD36 Expression.

Retinoid X receptor alpha (RXRA) is a well-characterized factor that regulates lipid metabolism; however, the regulatory mechanism in muscle cells of poultry is still unknown. The overexpression and the knockdown of RXRA in myoblasts (CS2 cells), RT-PCR, and western blotting were used to detect the expression levels of genes and proteins related to PPAR-signaling pathways. Intracellular triglycerides (TGs), cholesterol (CHOL), and nonesterified free fatty acids (NEFAs) were detected by the Elisa kit. Fat droplets were stained with Oil Red O. The double-fluorescein reporter gene and chromatin immunoprecipitation (CHIP) were used to verify the relationship between RXRA and candidate target genes. The RXRA gene was highly expressed in duck breast muscle, and its mRNA and its protein were reduced during the differentiation of CS2 cells. The CS2 cells, with the overexpression of RXRA, showed reduced content in TGs, CHOL, NEFAs, and lipid droplets and upregulated the mRNA expression of CD36, ACSL1, and PPARG genes and the protein expression of CD36 and PPARG. The knockdown of RXRA expression in CS2 cells enhanced the content of TGs, CHOL, NEFAs, and lipid droplets and downregulated the mRNA and protein expression of CD36, ACLS1, ELOVL6, and PPARG. The overexpression of the RXRA gene, the activity of the double-luciferase reporter gene of the wild-type CD36 promoter was higher than that of the mutant type. RXRA bound to -860/-852 nt, -688/-680 nt, and -165/-157 nt at the promoter region of CD36. Moreover, the overexpression of CD36 in CS2 cells could suppress the content of TGs, CHOL, NEFAs, and lipid droplets, while the knockdown expression of CD36 increased the content of TGs, CHOL, NEFAs, and lipid droplets. In this study, the transcription factor, RXRA, inhibited the accumulation of TGs, CHOL, NEFAs, and fat droplets in CS2 cells by promoting CD36 expression.

Identification of key genes and pathways associated with feed efficiency of native chickens based on transcriptome data via bioinformatics analysis.

BACKGROUND: Improving feed efficiency is one of the important breeding targets for poultry industry. The aim of current study was to investigate the breast muscle transcriptome data of native chickens divergent for feed efficiency. Residual feed intake (RFI) value was calculated for 1008 closely related chickens. The 5 most efficient (LRFI) and 5 least efficient (HRFI) birds were selected for further analysis. Transcriptomic data were generated from breast muscle collected post-slaughter. RESULTS: The differently expressed genes (DEGs) analysis showed that 24 and 325 known genes were significantly up- and down-regulated in LRFI birds. An enrichment analysis of DEGs showed that the genes and pathways related to inflammatory response and immune response were up-regulated in HRFI chickens. Moreover, Gene Set Enrichment Analysis (GSEA) was also employed, which indicated that LRFI chickens increased expression of genes related to mitochondrial function. Furthermore, protein network interaction and function analyses revealed ND2, ND4, CYTB, RAC2, VCAM1, CTSS and TLR4 were key genes for feed efficiency. And the 'phagosome', 'cell adhesion molecules (CAMs)', 'citrate cycle (TCA cycle)' and 'oxidative phosphorylation' were key pathways contributing to the difference in feed efficiency. CONCLUSIONS: In summary, a series of key genes and pathways were identified via bioinformatics analysis. These key genes may influence feed efficiency through deep involvement in ROS production and inflammatory response. Our results suggested that LRFI chickens may synthesize ATP more efficiently and control reactive oxygen species (ROS) production more strictly by enhancing the mitochondrial function in skeletal muscle compared with HRFI chickens. These findings provide some clues for understanding the molecular mechanism of feed efficiency in birds and will be a useful reference data for native chicken breeding.

Genome-wide genetic structure and selection signatures for color in 10 traditional Chinese yellow-feathered chicken breeds.

BACKGROUND: Yellow-feathered chickens (YFCs) have a long history in China. They are well-known for the nutritional and commercial importance attributable to their yellow color phenotype. Currently, there is a huge paucity in knowledge of the genetic determinants responsible for phenotypic and biochemical properties of these iconic chickens. This study aimed to uncover the genetic structure and the molecular underpinnings of the YFCs trademark coloration. RESULTS: The whole-genomes of 100 YFCs from 10 major traditional breeds and 10 Huaibei partridge chickens from China were re-sequenced. Comparative population genomics based on autosomal single nucleotide polymorphisms (SNPs) revealed three geographically based clusters among the YFCs. Compared to other Chinese indigenous chicken genomes incorporated from previous studies, a closer genetic proximity within YFC breeds than between YFC breeds and other chicken populations is evident. Through genome-wide scans for selective sweeps, we identified RALY heterogeneous nuclear ribonucleoprotein (RALY), leucine rich repeat containing G protein-coupled receptor 4 (LGR4), solute carrier family 23 member 2 (SLC23A2), and solute carrier family 2 member 14 (SLC2A14), besides the classical beta-carotene dioxygenase 2 (BCDO2), as major candidates pigment determining genes in the YFCs. CONCLUSION: We provide the first comprehensive genomic data of the YFCs. Our analyses show phylogeographical patterns among the YFCs and potential candidate genes giving rise to the yellow color trait of the YFCs. This study lays the foundation for further research on the genome-phenotype cross-talks that define important poultry traits and for formulating genetic breeding and conservation strategies for the YFCs.

Expression of genes related to lipid transport in meat-type ducks divergent for low or high residual feed intake.

OBJECTIVE: This study examined the effects of divergence in residual feed intake (RFI) on expression profiles of key genes related to lipid transport in the liver and duodenal epithelium and their associations with feed efficiency traits in meat-type ducks. METHODS: A total of 1,000 male ducks with similar body weight (1,042.1±87.2 g) were used in this study, and their individual RFI was calculated from 21 to 42 d of age. Finally, the 10 highest RFI (HRFI) and 10 lowest RFI (LRFI) ducks were chosen for examining the expression of key genes related to lipid transport in the liver and duodenal epithelium using quantitative polymerase chain reaction. RESULTS: In the liver, expression levels of albumin (ALB), CD36 molecule (CD36), fatty acid hydroxylase domain containing 2 (FAXDC2), and choline kinase alpha (CHKA) were significantly higher in LRFI ducks than in HRFI ducks (p<0.01); negative correlations (p<0.05) between expression levels of ALB, CD36, FAXDC2, and CHKA and RFI were detected in the liver. Additionally, ALB expression was strongly positively correlated (p<0.05) with CD36, FAXDC2, CHKA, and apolipoprotein H (APOH) expression in the liver. In duodenal epithelium, we found that mRNA levels of ALB, CD36, FAXDC2, and APOH were significantly higher in LRFI ducks than in HRFI ducks (p<0.01); RFI was strongly negatively correlated (p<0.05) with ALB, FAXDC2, and APOH expression, while ALB expression was strongly positively correlated with APOH expression (p<0.01) in duodenal epithelium. Furthermore, expression levels of both ALB and FAXDC2 genes were significantly associated with feed conversion ratio and RFI in both liver and duodenal epithelium (p<0.05). CONCLUSION: Our findings therefore suggest that ALB and FAXDC2 genes might be used as potential gene markers designed to improve feed efficiency in future meat-type duck breeding programs.

Association analysis between feed efficiency and expression of key genes of the avTOR signaling pathway in meat-type ducks.

Genes involved in the target of rapamycin (TOR) signaling pathway are implicated in nutrient translation, cell proliferation and differentiation, and anabolism, which can affect both growth and feed intake. However, the role of TOR signaling in the regulation of feed intake and feed efficiency in poultry is not clear. In the present study, a total of 1000 ducks, of similar initial weight, were chosen and transferred to individual cages to determine their residual feed intake (RFI) from the age of 21 to 42 days. Subsequently, 60 ducks, which were divided into high (HRFI) and low (LRFI) groups according to their RFI, were chosen to analyze the TOR signaling activities in the liver. The differential expression level of genes involved in the TOR signaling pathway was assayed by the real-time polymerase chain reaction. In the liver, the expression of AKT, avTOR, avLST8, and S6K1 was significantly higher in LRFI ducks than in HRFI ducks; avTOR and AKT were negatively associated with the feed conversion ratio and RFI. Furthermore, PI3K was moderately positively associated with AKT; AKT was strongly positively associated with PI3K, avTOR, avLST8, and S6K1; and avTOR was strongly positively associated with S6K1. In conclusion, the activation of avTOR signaling in the liver of LRFI ducks might be ascribed to higher energy state or more active nutrient transport (amino acids), or both, than those in the liver of HRFI ducks. The results of the present study indicate that AKT and avTOR of TOR signaling might be used as candidate genes to assess molecular regulation of feed efficiency.

Association of polymorphisms in Pit-1 gene with growth and feed efficiency in meat-type chickens.

OBJECTIVE: The pituitary specific transcription factor-1 (Pit-1) gene is responsible for pituitary development and growth hormone expression and is regarded as a pivotal candidate gene for growth and production in chickens. Therefore, the aim of this study was to investigate the association of Pit-1 polymorphisms with growth and feed efficiency traits in yellow meat-type chickens. METHODS: In the present study, five single nucleotide polymorphisms (SNPs) of Pit-1 were selected and genotyped by high-throughput matrix-assisted laser desorption-ionization time-of-flight mass spectrometry in 724 meat-type chickens. RESULTS: Association analysis showed that rs13687126 of Pit-1 was strongly associated with body weight gain (BWG) and feed intake (FI) (p<0.05), and that rs13687128 was significantly correlated with body weight at 70 days of age (BW70), BWG and feed conversion ratio (FCR) (p<0.05). SNP rs13905622 was strongly related to BW70 and FCR (p<0.05). Furthermore, birds with the GG genotype of rs13687126 had larger BWG and FI than those with the AG genotype (p<0.05). Individuals with the TT genotype of rs13687128 were significantly higher BW70 and BWG than those of the CT and CC genotype, while FCR was just the opposite (p<0.05). For rs13905622, the AA chickens showed strongly larger BW70 and lower FCR compared with the AT and TT chickens (p<0.05). Additionally, an ACA haplotype based on rs13687126, rs13687128, and rs13905622 had significant effects on BW70 and FCR (p<0.05). CONCLUSION: Our studies thus provide crucial evidence for the relationship between polymorphisms of Pit-1 and growth and feed efficiency traits which may be useful for meat-type chicken breeding programs.

Polymorphisms in the uncoupling protein 3 gene and their associations with feed efficiency in chickens.

OBJECTIVE: The uncoupling protein 3 (UCP3) is a member of the mitochondrial anion carrier superfamily and has crucial effects on growth and feed efficiency in many species. Therefore, the objective of the present study was to examine the association of polymorphisms in the UCP3 gene with feed efficiency in meat-type chickens. METHODS: Six single nucleotide polymorphisms (SNPs) of the UCP3 gene were chosen to be genotyped using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry in meat-type chicken populations with 724 birds in total. Body weight at 49 (BW49) and 70 days of age (BW70) and feed intake (FI) in the interval were collected, then body weight gain (BWG) and feed conversion ratio (FCR) were calculated individually. RESULTS: One SNP with a low minor allele frequency (<1%) was removed by quality control and data filtering. The results showed that rs13997809 of UCP3 was significantly associated with BWG and FCR (p<0.05), and that rs13997811 had significant effects on BW70 and BWG (p<0.05). Rs13997812 of UCP3 was strongly associated with BW70, FI, and FCR (p<0.05). Furthermore, individuals with AA genotype of rs13997809 had significantly higher BWG and lower FCR (p<0.05) than those with AT genotype. The GG individuals showed strongly higher BW70 and BWG than AA birds in rs13997811 (p<0.05). Birds with the TT genotype of rs13997812 had significantly greater BW70 and lower FCR compared with the CT birds (p<0.05). In addition, the TAC haplotype based on rs13997809, rs13997811, and rs13997812 showed significant effects on BW70, FI, and FCR (p<0.05). CONCLUSION: Our results therefore demonstrate important roles for UCP3 polymorphisms in growth and feed efficiency that might be used in meat-type chicken breeding programs.

Associations of polymorphisms in GHRL, GHSR, and IGF1R genes with feed efficiency in chickens.

The ghrelin (GHRL), ghrelin receptor (GHSR), and insulin-like growth factor 1 receptor (IGF1R) genes have crucial effects on body weight (BW), body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) in many species. However, few studies on associations of GHRL, GHSR, and IGF1R with BWG, FI, and FCR have been reported in chickens. In this study, 16 SNPs in GHRL, GHSR, and IGF1R genes were genotyped by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The objective of this study was to examine the associations of GHRL, GHSR, and IGF1R genes polymorphisms with BW at 49 days (BW49) and 70 days (BW70) of age, BWG, FI, and FCR in the interval in two yellow meat-type populations with a total of 724 birds. The results showed that rs15675067 of GHRL was significantly associated with BW70, BWG, and FCR (P < 0.05). For GHSR, rs16675844 had significant effects on FI and FCR (P < 0.01), and that rs14678932 showed significant association with BWG and FI (P < 0.05). Rs14011780 of IGF1R was strongly associated with BW49, BW70, and FCR (P < 0.05). Furthermore, haplotypes based on three SNPs of rs14986828, rs15675067, and rs15675065 in GHRL were significantly associated with BW70 and FCR (P < 0.05). Meanwhile, a three-SNP haplotype comprising rs14011783, rs14011780, and rs14011776 in IGF1R showed significant effects on BW49, BW70, and FCR (P < 0.05). Therefore, it was concluded that the identified SNPs and analyzed haplotypes in this study might be useful for broiler breeding programs.

Association of leptin receptor gene polymorphisms with growth and feed efficiency in meat-type chickens.

Many genetic factors influence the growth and feed intake of birds. In the current study, we evaluated the association of 8 previously reported SNP in the chicken leptin receptor (LEPR) gene with BW, BW gain (BWG), feed intake (FI), and feed conversion ratio (FCR). Four SNP with a very low minor allele frequency were removed by genotype quality control. The experimental population consisted of 796 pedigreed males from 2 genetically unrelated yellow meat-type chicken strains, 335 chickens from N202, and 461 chickens from N301. The BW at 49 (BW49) and 70 d of age (BW70) and FI (from 49 to 70 d of age) were measured individually. The BWG and FCR were calculated based on BW and FI in the interval between 49 to 70 d. The results indicated that rs14657336, rs13684613, rs13684615, and rs13684616 were found in strong linkage disequilibrium. This linkage disequilibrium block was significantly associated with BW49 (P < 0.05), BW70 (P < 0.05), and FI (P < 0.05) in the N202 strain, and FCR (P < 0.01) in the N301 strain, respectively. In addition, the GTACGTAC diplotype had the highest BWG and FI in both strains. The association revealed in this study suggests the need for further functional study on the role of LEPR gene in regulating feed intake and FCR of chickens.

Prevalence and molecular characterization of Trichomonas gallinae from pigeons in Anhui, China.

Trichomonas gallinae, a protozoan parasite causing avian trichomonosis, exhibits a widespread global prevalence. It primarily affects the upper digestive tract of birds and has resulted in significant ecological problems worldwide. This study aimed to investigate the prevalence and genotypes of T. gallinae in Anhui Province, China. A total of 1612 oropharyngeal swab samples were collected from pigeon farms in Anhui Province to determine the prevalence of T. gallinae infection. The results revealed 565 (35.1%) positive samples of T. gallinae. Significant differences in infection rates were observed among different regions and age groups. Furthermore, the ITS1/5.8 S/ITS2 region was amplified, sequenced, and subjected to phylogenetic analysis. Genotypes A and B of T. gallinae were identified, and genotype B was the dominant genotype in Anhui Province. This is the first report on the prevalence and molecular characterization of T. gallinae in Anhui Province, China. Additionally, we integrated reports on the prevalence and genotype of T. gallinae in relevant provinces in China.

Polymorphisms in the transforming growth factor ß3 gene and their associations with feed efficiency in chickens.

Growth and feeding traits such as BW, BW gain (BWG), feed intake (FI), and feed conversion ratio (FCR) are of economic importance in poultry production. In this study, 8 SNP of the transforming growth factor ß3 (TGF-ß3) gene, which are located in the proximity of quantitative trait loci affecting BW and FCR, were selected to be genotyped by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in 2 yellow meat-type chicken populations with 724 birds in total. Body weights at 49 (BW49) and 70 (BW70) d of age and FI in the interval were recorded, and respective BWG and FCR were calculated for each bird. Two SNP with a very low minor allele frequency (<1%) were discarded from further analysis. The results showed that both rs13586818 and rs14535174 had significant effects (P < 0.05) on BWG and FCR, and that rs14535177 was significantly (P < 0.05) associated with BW49, BW70, and FI. Furthermore, birds with the GA genotype of rs13586818 showed strongly higher FCR and lower BWG (P < 0.05) compared with AA individuals. The TT birds had significantly larger BWG (P < 0.05) than GT birds in rs14535174, whereas FCR was the opposite. Individuals with the GC genotype of rs14535177 had significantly higher BW49, BW70, FCR, and FI (P < 0.05) than those with the GG genotype. Additionally, haplotypes based on 3 SNP of rs13586818, rs14535174, and rs14535177 were significantly associated with FCR (P < 0.05). The SNP and analyzed haplotypes identified in this study might be used as potential genetic markers in meat-type chicken breeding.

The Duck RXRA Gene Promotes Adipogenesis and Correlates with Feed Efficiency.

BACKGROUND: The accumulation of fat in ducks is the main cause of low feed efficiency and metabolic diseases in ducks. Retinoic acid X receptor alpha (RXRA) is a member of the nuclear receptor superfamily involved in lipid, glucose, energy, and hormone metabolism. The effect of the RXRA gene on lipid metabolism in duck preadipocytes (DPACs) and the relationship between SNPs and the feed efficiency traits of ducks are unclear. METHODS: qRT-PCR and Western blotting analyses were used to detect changes in mRNA and protein in cells. Intracellular triglycerides (TGs) were detected using an ELISA kit. A general linear model analysis was used to determine the association between RXRA SNPs and feed efficiency. RESULTS: The duck RXRA gene was highly expressed on the fourth day of DPAC differentiation. The RXRA gene increased the content of fat and TG in DPACs and promoted the expression of cell differentiation genes; g.5,952,667 correlated with average daily feed intake (ADFI), residual feed intake (RFI), and feed conversion ratio (FCR). CONCLUSIONS: Duck RXRA can accelerate fat accumulation, and the polymorphism of the RXRA gene is closely related to feed efficiency, which provides basic data for breeding high feed efficiency ducks.

Next-Generation Sequencing of the Complete Huaibei Grey Donkey Mitogenome and Mitogenomic Phylogeny of the Equidae Family.

The Huaibei grey donkey (HGD) is an endangered species and a vital native breed in Anhui Province, China. However, its complete mitogenome, phylogeny, and maternal origin remain unclear. The objectives of this study were to detect the genetic diversity of the HGD and investigate its phylogenetic relationship with other breeds to inform conservation management. The complete mitogenome of the HGD was sequenced through next-generation sequencing, and the most variable region in the mitochondrial DNA displacement-loop (D-loop) was amplified via a polymerase chain reaction (PCR). Next, we used the median-joining network (MJN) to calculate the genetic relationships among populations and the neighbor-jointing method to build a phylogenetic tree and speculate as to its origin. The results showed that the mitogenome contains 22 tRNAs, 2 rRNAs, 13 PCGs, and 1 D-loop region. Analyzing the D-loop region of the HGDs, we identified 23 polymorphic sites and 11 haplotypes. The haplotype and nucleotide diversity were 0.87000 (Hd) and 0.02115 (Pi), respectively. The MJN analysis indicated that the HGD potentially has two maternal lineages, and phylogenetic analysis indicated that the Somali lineage could be the most probable domestication center for this breed. Therefore, our mitogenome analysis highlights the high genetic diversity of the HGD, which may have originated from the Somali wild ass, as opposed to the Asian wild ass. This study will provide a useful resource for HGD conservation and breeding.

Mitochondrial genome and phylogenetic analysis of Chaohu duck.

A complete mitochondrial genome sequence is important for the accurate determination of phylogenetic relationships. Chaohu duck is a dominant native breed in Anhui Province, China. We aimed to ascertain the complete mitochondrial genome sequence of Chaohu duck via high-throughput sequencing and primer walking. Phylogenetic analysis of Chaohu duck was performed following Kimura 2-parameter model. The total length of the mitogenome was 16,597 bp, and comprised 29.2 %A, 22.2 % T, 32.8 % C, and 15.8 % G. It included 2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA genes and a control region (D-loop). Furthermore, the haplotype diversity and nucleotide diversity values were 0.9028(Hd) and 0.01162(Pi) respectively. This indicates that Chaohu duck has high population diversity. Twenty-two haplotypes were identified in sixty Chaohu ducks which were divided into two haplogroups. Therefore, we inferred that Chaohu duck may originate from Anas platyrhynchos, and was influenced by Anas poecilorhyncha during evolution. Our results provide mitochondrial genome information for further studies on Chaohu ducks and lays a foundation for germplasm resources conservation.

Impact of divergence of residual feed intake on triglyceride metabolism-related gene expression in meat-type ducks.

Triglyceride (TG) metabolism is a key factor that affects residual feed intake (RFI); however, few studies have been conducted on the related gene expression in poultry. The aim of the present study was to investigate the expression of genes and their associations with RFI in meat-type ducks. Weight gain and feed intake (FI) at an age 21-42 days were measured and the RFI was calculated. Quantitative PCR was used to test the expression of the six identified genes, namely peroxisome proliferator activated receptor γ (PPARγ), glycerol kinase 2 (GK2), glycerol-3-phosphate dehydrogenase 1 (GPD1), glycerol kinase (GYK), lipase E (LIPE), and lipoprotein lipase (LPL) in the duodenum in the high RFI (HRFI) and low RFI (LRFI) groups. The results demonstrated that daily feed intake, feed conversion ratio (FCR), and RFI were markedly higher in HRFI ducks than those in LRFI ducks. Moreover, the levels of expression of PPARγ, GK2, and LIPE were significantly higher in the LRFI group than those in the HRFI group. Correlation analysis showed that PPARγ, GK2, and LIPE were significantly negatively associated with FCR and RFI. Furthermore, gene expression levels were negatively associated with the measured phenotype. The association of GK2 with PPARγ, GPD1, LPL, and LIPE was positive. The relationship between the TG related gene and RFI was further verified to potentially develop pedigree poultry breeding programs. The results of this study suggested that the expression of genes correlated with TG metabolism and transport is up-regulated in the duodenum of ducks with high feed efficiency. PPARγ, GK2, and LIPE are important genes that affect RFI. The results of the present study provide information that could facilitate further explorations of the mechanism of RFI and potential markers at the molecular and cellular levels.

Characteristics of the complete mitochondrial genome of Douhua chicken (Gallus gallus) and phylogenetic considerations.

Douhua chicken is a unique local breed from Anhui Province, China. This study aimed to illustrate the Douhua chicken mitogenome and clarify its phylogenetic status by sequencing and annotating the complete mitochondrial genome using high-throughput sequencing and primer walking. Phylogenetic analysis through the Kimura 2-parameter model indicated the maternal origin of Douhua chicken. The results revealed that the mitochondrial genome is a closed circular molecule (16,785 bp) that consists of 13 protein-coding genes, 22 transfer RNA (tRNA) coding genes, two ribosomal RNA (rRNA) coding genes, and a control region. The base composition of the Douhua chicken mitogenome contains 30.3% A, 23.7% T, 32.5% C, and 13.5% G, and the haplotype and nucleotide diversity values are 0.829 (Hd) and 0.00441 (Pi), respectively. Furthermore, 10 haplotypes of D-loop sequences among 60 Douhua chickens were identified and distributed into four haplogroups (A, C, D, and E). Overall, the result of the present study indicates that Douhua chicken may have originated from Gallus gallus, and this process was influenced by Gallus gallus spadiceus, Gallus gallus murghi, and Gallus gallus bankiva. This study provides novel mitogenome data to support further phylogenetic and taxonomic studies on Douhua chicken. Additionally, the findings of this study will provide deeper insights for identifying the genetic relationships among populations and tracing maternal origins based on phylogenetic considerations for use in studies on the geographic conservation, utilization, and molecular genetics of poultry species.

In the modern poultry industry, resources of native varieties have become major aspects. Douhua chicken is a medium-sized, slow-growing, and white-feathered local breed that represents a popular local chicken breed in Anhui Province, China. This breed is adaptable and exhibits important production traits and a stable inheritable characteristics, such as delicious meat and stable egg-laying performance. The present study aimed to provide a better understanding of the germplasm characteristics and phylogenetic relationships of Douhua chicken by analyzing its complete mitochondrial genome sequence and a describing its genomic composition, nucleotide composition, and gene structure. The present study provides theoretical support for the protection, development, and utilization of Douhua chicken resources. Additionally, this study provides new mitochondrial genome data to support further phylogenetic and taxonomic studies conducted on Douhua chicken.

Research Note: Genetic parameters for egg production and clutch-related traits in indigenous Beijing-You chickens.

Egg products from indigenous chickens have growing market shares as consumers are pursuing differentiation in egg consumption. The genetic improvement in egg production performance of those breeds is crucial for increasing the economic profit. This study aimed to estimate genetic parameters for egg production and clutch-related traits in indigenous Beijing-You chickens for understanding the genetic architecture and exploring proper biological traits for selection. Data on traits including age at first egg (AFE), egg number (EN), average clutch length (ACL), maximum clutch length (MCL), number of clutches (NC) and pauses (NP), and average pause length (APL) were collected from 4 generations of purebred Beijing-You chickens based on the 43-wk and 66-wk of individual egg production record. The heritabilities, genetic and phenotypic correlations were analyzed by the DMU software with the restricted maximum likelihood method in a multivariate animal model. The results showed that the AFE of Beijing-You chickens was 174.45 d of age, and its heritability was as high as 0.62. The heritability was 0.26 for EN43 and 0.18 for EN66. The clutch traits including ACL, MCL, NC, and NP were moderate to high heritable (h2 = 0.15-0.39), but APL was very low heritable (h2 = 0.05). Genetic correlations were high between AFE and EN (rG(AFE, EN43) = -0.79, rG(AFE, EN66) = -0.39), whereas low between AFE and ACL (rG(AFE, ACL43) = -0.08, rG(AFE, ACL66) = 0.01) and MCL (rG(AFE, MCL) = -0.07). EN had higher correlations with ACL (rG(EN43, ACL43) = 0.59, rG(EN66, ACL66) = 0.40) than that with MCL (rG(EN43, MCL43) = 0.56, rG(EN66, MCL66) = 0.32). The heritability for ACL43 (h2 = 0.38) was higher than that for MCL43 (h2 = 0.33). ACL43 had a positive correlation with EN66 (rG(ACL43, EN66) = 0.62). These results indicated that the egg production of whole laying period could be improved by early selection for AFE and ACL at the same time in Beijing-You chickens.

Identification of stable reference genes for quantitative gene expression analysis in the duodenum of meat-type ducks.

Quantitative polymerase chain reaction (qPCR) is an important method to detect gene expression at the molecular level. The selection of appropriate housekeeping genes is the key to accurately calculating the expression level of target genes and conducting gene function studies. In this study, the expression of eight candidate reference genes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-actin (ß-actin), 18S ribosomal RNA (18S rRNA), hydroxymethylbilane synthase (HMBS), hypoxanthine phosphoribosyltransferase 1 (HPRT1), TATA box binding protein (TBP), ribosomal protein L13 (RPL13), and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein (YWHAZ), in the duodenal epithelial tissue of 42-day-old meat-type ducks were detected using qPCR. Furthermore, their expression stability was analyzed using the geNorm, NormFinder, and BestKeeper programs. The results indicated that HMBS and YWHAZ were the most stably expressed genes. All three programs indicated that the expression of 18S rRNA was the least stable, making it unsuitable for the study of gene expression in meat-type duck tissues. This study provides stable reference genes for gene expression analysis and contributes to further studies on the gene function of meat-type ducks.

Complete Mitochondrial Genome, Genetic Diversity and Phylogenetic Analysis of Pingpu Yellow Chicken (Gallus gallus).

In this study, the complete mitochondrial genome sequence of one female Pingpu Yellow chicken (PYC) and the D-loop sequences obtained from 60 chickens were analyzed to investigate their genetic diversity and phylogeny. The total length of the PYC mitogenome is 16,785 bp and that of the complete D-loop is 1231 to 1232 bp. The mitogenome comprises 22 transfer ribonucleic acids (tRNAs), 2 ribosomal ribonucleic acids (rRNAs), 13 protein-coding genes (PCGs), and 1 non-coding control region (D-loop). Additionally, the total length of the 13 PCGs is 11,394 bp, accounting for 67.88% of the complete mitogenome sequence, and the PCGs region has 3798 codons. A majority of the PCGs have ATG as the start codon. The haplotype and nucleotide diversity of PYC were 1.00000 ± 0.00029 and 0.32678 ± 0.29756, respectively. In the D-Loop data set, we found 25 polymorphic sites, which determined 18 haplotypes and 3 major haplogroups (A-C). Therefore, PYC has a classical vertebrate mitogenome, with comparatively high nucleotide diversity and potentially three maternal lineages. The neighbor-joining (NJ) tree analysis results showed PYC grouped with the Luhua (MT555049.1) and Nandan chickens (KP269069.1), which indicates that PYC is closely related to these two breeds.

Complete mitochondrial genome and phylogenetic analysis of Huangshan Black chicken (Gallus gallus).

In this study, we sequenced the complete mitochondrial genome (mitogenome) of the Huangshan Black chicken (HBC). Results showed that the complete HBC mitogenome was 16,785 bp in size, comprising 22 transfer RNA genes, 2 ribosomal RNA genes, 13 protein-coding genes, and 1 non-coding control region (D-loop). The overall nucleotide composition was 32.5% for C, 30.3% for A, 23.7% for T, and 13.5% for G. Phylogenetic analysis showed that the HBC mitogenome was clustered with Xianju chicken, which belonged to the haplogroup D2. Our results therefore demonstrate that the origin of HBC corresponds to haplogroup D2 distribution and might have at least one maternal lineage originated from Southeast Asia.