鸡蛔虫线粒体cox1和nad4基因的遗传变异分析

doi:10.16431/j.cnki.1671-7236.2017.01.016

摘要/Abstract

摘要：

为探究鸡蛔虫的种内遗传变异和系统发育关系，对分离自四川省西昌市的15个鸡蛔虫分离株的线粒体细胞色素c氧化酶第Ⅰ亚基（cox1）基因部分序列和烟酰胺腺嘌呤二核苷酸脱氢酶亚单位Ⅳ（nad4）基因全序列进行PCR扩增、序列测定及分析，并用cox1和nad4基因部分序列构建鸡蛔虫与其他蛔虫的NJ树和贝叶斯树。测序结果显示，所获得的鸡蛔虫cox1和nad4基因全序列长度分别为1 152和1 236 bp，分别有33和40个变异位点，碱基变异率分别为0~2.1%和0~2.3%；分别检测到8和11个单倍型，总的单倍型多样性分别为0.733±0.124和0.933±0.054，核苷酸多样性分别为0.00433±0.00153和0.00541±0.00157，平均遗传距离分别为0.004和0.005。构建的种系发育树均显示15个鸡蛔虫西昌分离株与其他国家/地区的鸡蛔虫分离株聚类形成一个分支，与鸽蛔虫的亲缘关系最近，与其他蛔虫所属的分支相隔较远。研究结果表明鸡蛔虫西昌分离株的遗传变异程度低，且nad4基因比cox1基因更适合作为研究鸡蛔虫分离株遗传变异的分子标记。

关键词: 鸡蛔虫; cox1基因; nad4基因; 遗传变异

Abstract:

To study the genetic variations of Ascaridia galli (A. galli) and the phylogentic relationships with other Ascaridata, fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and complete sequence of NADH dehydrogenase subunit 4 (nad4) gene of 15 adult A. galli isolated from Xichang city in Sichuan province were amplified by PCR, then analyzed the sequences. NJ trees and Bayes trees based on pcox1 and pnad4 genes were reconstructed. The partial sequences of cox1 gene and complete sequence of nad4 gene were 1 152 and 1 236 bp, respectively. There were 33 and 40 variable sites in the pcox1 and nad4 gene sequences, the intraspecific sequence variations within A. galli were 0 to 2.1% for pcox1 gene, 0 to 2.3% for nad4 gene. 8 and 11 haplotypes were detected in pcox1 and nad4 genes, the global haplotype diversity were 0.733±0.124 and 0.933±0.054, the nucleotide diversities were 0.00433±0.00153 and 0.00541±0.00157, and the average genetic distances were 0.004 and 0.005, respectively. Phylogenetic trees based on pcox1 and pnad4 genes with Neighbour-Joining and Bayesian analysis method revealed that all A. galli were clustered in one clade, and they were more closely related to A. columbae than any other members of Ascaridata. These findings indicated that 15 isolates of A. galli from Xichang city kept low genetic variation, nad4 gene was more suitable and effective than cox1 gene as molecular marker for detecting genetic variation of A. galli.

Key words: Ascaridia galli; cox1 gene; nad4 gene; genetic variation

中图分类号:

S813.3

郝桂英, 何学谦, 万润. 鸡蛔虫线粒体cox1和nad4基因的遗传变异分析[J]. 《中国畜牧兽医》, 2017, 44(1): 113-122.

HAO Gui-ying, HE Xue-qian, WAN Run. Genetic Variation Analysis of Mitochondrial cox1 and nad4 Genes of Ascaridia galli[J]. , 2017, 44(1): 113-122.

参考文献

[1] 杨光友.动物寄生虫病学[M].第3版.成都:四川科学技术出版社,2009.
[2] Abdelqder A,Gauly M,Wollny B A,et al.Prevalence and burden of gastrointestinal helminthes among local chickens, in Northern Jordan[J].Prev Vet Med,2008,85(1-2):17-22.
[3] Jansson D S,Nyman A,Vagsholm I,et al.Ascarid infections in laying hens kept in different housing systems[J].Avian Pathol,2010,39(6):525-532.
[4] 贾万忠,闫鸿斌,倪兴维,等.线虫线粒体基因组全系列分析研究进展[J].中国农业科学,2011,44(6):1255-1265.
[5] Xie Y,Niu L,Zhao B,et al.Complete mitochondrial genomes of chimpanzee- and gibbon- derived Ascaris isolated from a zoological garden in Southwest China[J].PLoS One,2013,8(12):e82795.
[6] 梁轩,刘毅.犬弓首蛔虫线粒体cox1基因的克隆及序列分析[J].中国动物检疫,2015,32(3):66-69.
[7] Chang Q C,Gao J F,Sheng Z H,et al.Sequence variability in three mitochondrial genes among four roundworm species from wild animals in China[J].Mitochondrial DNA,2015,26(1):75-78.
[8] 宋美冉,李康信,石先利,等.华南虎狮弓蛔虫三种线粒体基因的遗传进化分析[J].中国动物传染病学报,2015,23(6):68-75.
[9] Mikaeili F,Mirhendi H,Mohebali M,et al.Sequence variation in mitochondrial cox1 and nad1 genes of Ascaridoid nematodes in cats and dogs from Iran[J].J Helminthol,2015,89(4):496-501.
[10] 萨姆布鲁克J,拉塞尔D W著.黄培堂,王嘉玺,朱厚础,等译.分子克隆实验指南[M].第3版.北京:科学出版社,2002.
[11] Katakam K K,Nejsum P,Kyvsgaard N C,et al.Molecular and parasitological tools for the study of Ascaridia galli population dynamics in chickens[J].Avian Pathol,2010,39(2):81-85.
[12] Li J Y,Liu G H,Wang Y,et al.Sequence variation in three mitochondrial DNA genes among isolates of Ascaridia galli originating from Guangdong,Hunan and Yunnan provinces,China[J].J Helminthol,2013, 87(3):371-375.
[13] Liu G H,Shao R F,Li J Y,et al.The complete mitochondrial genomes of three parasitic nematodes of birds:A unique gene order and insights into nematode phylogeny[J].BMC Genomics,2013,14(1):414.
[14] Liu G H,Wu C Y,Song H Q,et al.Comparative analyses of the complete mitochondrial genomes of Ascaris lumbricoides and Ascaris suum from humans and pigs[J].Gene,2012,492(1):110-116.
[15] Li M W,Lin R Q,Song H Q,et al.The complete mitochondrial genomes for three Toxocara species of human and animal health significance[J].BMC Genomics,2008,9(1):224.
[16] Wickramasinghe S, Yatawara L, Rajapakse R P, et al.Toxocara vitulorum (Ascaridida:Nematoda):Mitochondrial gene content,arrangement and composition compared with other Toxocara species[J]. Mol Biochem Parasitol, 2009,166(1):89-92.
[17] Liu G H,Zhou D H,Zhao L,et al.The complete mitochondrial genome of Toxascaris leonina:Comparison with other closely related species and phylogenetic implications[J].Infect Genet Evol,2014,21(1):329-333.
[18] Xie Y,Zhang Z,Wang C,et al.Complete mitochondrial genomes of Baylisascaris schroederi, Baylisascaris ailuri and Baylisascaris transfuga from giant panda,red panda and polar bear[J]. Gene, 2011,482(1-2):59-67.
[19] Xie Y, Zhang Z, Niu L, et al. The mitochondrial genome of Baylisascaris procyonis[J].PLoS One, 2011, 6(10):e27066.
[20] Lavrov D V, Brown W M. Trichinella spiralis mtDNA:A nematode mitochondrial genome that encode a putative ATP8 and normally structured tRNAs and has a gene arrangement relatable to those of coelomate metazoans[J].Genetics,2001,157(2):621-637.
[21] 伍慧兰,谭美英,刘国华,等.湖南省鸡蛔虫线粒体cox1和nad1基因的序列测定及种系发育分析[J].畜牧兽医学报,2010,41(12):1592-1597.
[22] 李佳缘,刘国华,王燕,等.鸡蛔虫线粒体cox1基因的克隆及序列分析[J].中国畜牧兽医,2012,39(5):41-43.
[23] 李淼,蔡丝丝,姚明锐,等.湖南省鸡蛔虫线粒体nad4基因序列的测定及分析[J].现代农业科技,2012,4:321.
[24] 李淼,侯强红,蔡丝丝,等.鸡蛔虫cox2和nad4 rDNA的序列测定及分析[J].中国兽医杂志,2013,49(1):19-21.