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Brazilian Journal of Poultry Science

Print version ISSN 1516-635XOn-line version ISSN 1806-9061

Braz. J. Poult. Sci. vol.21 no.1 Campinas  2019  Epub Apr 18, 2019

https://doi.org/10.1590/1806-9061-2018-0784 

ORIGINAL ARTICLES

The Complete Mitochondrial Genome of the Donganyellow Chicken and Its Phylogenetic Analyses

IResearch Department of Animal Nutrition and Poultry, Hunan Institute of Animal and Veterinary Science, Changsha, PR China

IIInstitute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha, PR China

IIICollege of Animal Science and Technology, Hunan Agricultural


ABSTRACT

Donganyellow chicken (Gallus gallusdomesticus, DYC) is one of the famous native breeds of Hunan province in China. It is the first time that the complete mitochondrialgenome sequence of DYC was reported. The total length of the mtDNA is16, 786bp. It contains 22transfer RNA genes, 2 ribosomal RNA genes,13 protein-coding genes and 1 D-loop region. The overall composition of the mtDNA is 30.27% for A, 23.74% for T, 32.50% for C and 13.49% for G. Phylogenetic analyses using N-J computational algorithms showed that the analyzed20Galliformes species are divided into three major clades: Phasianidae, Numidiidae and Odontophoridae. In addition, our work confirmed that DYCand Taoyuan chickenhave a closegenetic relationship with fellow tribal members Xuefeng black-boned chicken and Huang Lang chicken. This work will provide an important data set for the study in genetic mechanism of chicken in Hunan province.

Keywords: Gallus gallusdomesticus; Donganyellow chicken; mitochondrial; phylogenetic analyses

INTRODUCTION

Donganyellow chicken (Gallus gallusdomesticus, DYC), is one of the important indigenous protected breeds of chicken in Hunan Province, which has been bred in Dongan county for hundreds of years. The features of DYC is strong adaptability, good stress resistance and good meat quality (Chen, 2013). In addition, domestic chickens (Gallus gallusdomesticus) play the keyrole in the agricultural and economic sectors in Hunan Province. There are numerous domestic chicken breeds in Hunan Province including Taoyuan chicken (TYC), Huang Lang chicken (HLC), Xuefeng black-boned chicken (XBC) and DYC. However, a better characterization of the complete mitochondrial genome and genetic diversity of four domestic chicken breeds mentioned above is in urgent need to increase the conservational application for these four species.

MATERIALS AND METHODS

Determination and analysis of the complete mitochondrial genome sequence of DYC

This study determined complete mitochondrial DNA sequence of DYC, which has been registered in GenBank under accession number KM886937. The chicken was farmed at Hunan Dongan Chicken Technology Co., Ltd. (Yongzhou City, Hunan Province, China). Mitochondrial DNA of DYC was extracted using the EasyPure Kit of Genomic DNA (Transgen Biotech, Beijing, China) according to its instruction manual. PCR was carried out to amplify the complete mitochondrial genome with 11 pairs of primers. PCR products of the Gel electrophoresis were purified by Gel AdvancedTM Gel Extraction (Rich Biotech, Taiwan, China) and sequenced by BioSune Biotech (Shanghai, China). The mitochondrial DNA sequence was analyzed using DNAStar.Lasergene.v7.1 software, tRNA Scan-SE1.21 software and DOGMA software.

Phylogenetic analysis

Phylogenetic analysis was performed using the complete mitochondrial DNA sequences of 20Galliformes. Each of the sequence dataset was aligned by ClustalXand analyzed by neighbor-joining (N-J) in MEGA 4.0, and bootstrap analysis was performed with 100 replications.

RESULTS AND DISCUSSION

The total length of the mtDNA is 16,786bp. It contains the typical structure, including 24 RNA genes, 13 protein-coding genes and 1 D-loop region as that of most other vertebrates (Lin et al. 2016a). The overall composition of the mtDNA was estimated to be 30.27% for A, 23.74% for T, 32.50% for C and 13.49% for G. All the protein initiation codons are ATG, except for COX1 which is GTG (Table 1). All the mtDNA genes were encoded on the heavy (H) strand except for eight types of tRNA genes and ND6 gene. All these genes have15 spaces in the length of 1-9bpand have 7 overlaps in the length of 1-10 bp. These genes had four types of termination codons, including TAA, TAG, AGG and ‘‘T- -’’, ‘‘T- -’’ is the 5’ terminal of the adjacent gene (Anderson et al. 1981). The lengths of 12S rRNA and the 16S rRNA were 975bp and 1623bp.Among 13 protein-coding genes, the longest one was ND5 gene (1818 bp) and the shortest one was ATPase8 gene (165bp). And deduced 22 tRNA genes were distributed in rRNA and protein-coding genes, ranging from 66 to 76 bp in size, and the situation was similar to other breeds of chicken in Hunan province(Yu et al. 2016; Liu et al. 2016a; Liu et al. 2016b). The D-loop region was located between tRNAGlu and tRNAPhe with a length of 1232bp. The putative origin for light strand replication (OLR) had been identified in the tRNATrp-tRNAAla-tRNAAsn-tRNACys-tRNATyr region (WANCY) of most vertebrates (Lin et al. 2016b; Wang et al. 2008). However, DYC might be associated with the deletion of OLR between tRNAAsn and tRNACys genes, which cannot be folded into a OLR-like secondary structure.

Table 1 Organization of the mitochondrial genome of Dongan yellow chicken. 

Gene name Position Size Codon Anti-codon Strand Space/overlap+
Start End Start Stop
D-loop 1 1232 1232
tRNA Phe 1233 1302 70 GAA H
12S rRNA 1303 2277 975 H
tRNA Val 2278 2350 73 TAC H
16S rRNA 2351 3973 1623 H
tRNA Leu 3974 4047 74 TAA H
ND1 4057 5031 975 ATG TAA H 9
tRNA Ile 5032 5103 72 GAT H
tRNA Gln 5109 5179 71 TTG L 5
tRNA Met 5179 5247 69 CAT H -1
ND2 5248 6288 1041 ATG TAG H
tRNA Trp 6287 6362 76 TCA H -2
tRNA Ala 6369 6437 69 TGC L 6
tRNA Asn 6441 6513 73 GTT L 3
tRNA Cys 6515 6580 66 GCA L 1
tRNA Tyr 6580 6650 71 GTA L -1
COX1 6652 8202 1551 GTG AGG H 1
tRNA Ser 8194 8268 75 TGA L -9
tRNA Asp 8271 8339 69 GTC H 2
COX2 8341 9024 684 ATG TAA H 1
tRNA Lys 9026 9093 68 TTT H 1
ATPase8 9095 9259 165 ATG TAA H 1
ATPase6 9250 9933 684 ATG TAA H -10
COX3 9933 10716 784 ATG T- - H -1
tRNA Gly 10717 10785 69 TCC H
ND3 10786 11137 352 ATG TAA H
tRNA Arg 11139 11206 68 TCG H 1
ND4L 11207 11503 297 ATG TAA H
ND4 11497 12874 1378 ATG T- - H -7
tRNA His 12875 12943 69 GTG H
tRNA Ser 12944 13010 67 AGA H
tRNA Leu 13011 13081 71 TAG H
ND5 13082 14899 1818 ATG TAA H
Cytb 14904 16046 1143 ATG TAA H 4
tRNA Thr 16050 16118 69 TGT H 3
tRNA Pro 16119 16188 70 TGG L
ND6 16195 16716 522 ATG TAA L 6
tRNA Glu 16719 16786 68 TTC L 2

T- - means incomplete termination codon. +Negative numbers indicate overlapping nucleotides.

An N-J tree showed that the analyzed species are divided into three major clades: Phasianidae, Numidiidae and Odontophoridae, moreover, the outgroup, Anseranser, is located at the base of the tree (Figure 1). Phasianidae makesup the first lineage, which is sister to the second group, Numidiidae; Odontophoridae forms the third group and is sister to Phasianidae and Numidiidae. This N-Jtree generated from the complete mitochondrial DNA for 20 Galliformes species have basically similar topologies. Three major branches are represented: Phasianidae, Acrylliumvulturinum and Colinus virginianus.The first lineage, subfamily Phasianidae, includes species of tribes Gallus, Bambusicola, Pavo, Tetraophasis, Meleagris, Phasianus, Chrysolophus and Coturnix. Acrylliumvulturinum is the sole member of the second group Numidiidae. The Phasianidae and Numidiidae are sister groups, which then group together with Colinus virginianus (Odontophoridae). Therefore, our work confirmed that DYC and TYC have a closegenetic relationship with fellow tribal members XBC and HLC (The overall average genetic distance between the complete mitochondrial genome sequences is 0.127). Meanwhile, we also found that DYC and TYC have highly similar genetic relationship which are consistent with those reported previously (Zhao et al. 2016).

Figure 1 Phylogenetic analysis based on complete mitochondrial genome sequences. An N-J tree was built based on the phylogenetic analysis of 20Galliformes species’ complete mitochondrial genomes. The mitochondrial genome sequences of the Galliformes species were obtained from the GenBank databases (Accession numbers have marked on the figure). Abbreviation of species indicates: DYC, Donganyellow chicken; TYC, Taoyuan chicken; XBC, Xuefeng black-bonedchicken; HLC, Huang Lang chicken. 

ACKNOWLEDGEMENTS

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article. This research was supported by International S&T Cooperation Program of China (ISTCP, 2013DFG31810) and Science & Technology Project of Hunan Province (HHN2017SC09).

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Received: March 23, 2018; Accepted: August 27, 2018

Corresponding author e-mail address Hai-Feng Yan Hunan Institute of Animal and Veterinary Science, No. 8 Changlang Road, Changsha City, Hunan Province, PR China. Email: hfyan2005@163.com

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