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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2014, Vol. 40 Issue (1): 103-110    DOI: 10.3785/j.issn.1008-9209.2013.06.012
动物科学     
东乡绿壳蛋鸡线粒体DNA控制区多态性及与9种地方乌鸡的进化分析
章学东1*， 李庆海1， 张成先2， 陈贤惠2， 楼立峰1， 王欢欢1
（1.杭州市农业科学研究院,杭州310024；2.浙江光大种禽业有限公司,浙江 海宁314423）
Mitochondrial DNA control region polymorphisms in Dongxiang blue-eggshell chicken and their evolutionary relatedness with nine domestic Wugu breeds
Zhang Xuedong1*, Li Qinghai1, Zhang Chengxian2, Chen Xianhui2, Lou Lifeng1, Wang Huanhuan1
(1. Hangzhou Academy of Agricultural Sciences, Hangzhou 310024, China; 2. Zhejiang Guangda Poultry Breeding Co. Ltd., Haining， Zhejiang 314423, China)
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摘要： 采用PCR产物直接测序的方法，对39只东乡绿壳蛋鸡（♂19、♀20）的线粒体DNA（mtDNA）控制区进行遗传多态性分析；并与9个国内其他地方乌鸡种的209个个体进行序列比对和进化分析。结果显示：东乡绿壳蛋鸡mtDNA控制区534 bp序列的G+C含量为43.1%；序列共存在21个变异位点，其中转换占71.4%（15/21）、颠换占28.6%（6/21）；核苷酸多样度为0.008 29±0.001 30。分析10个乌鸡种的248个个体共获得73种单倍型，其中东乡绿壳蛋鸡含9种单倍型，单倍型多样度为0.773±0.048。品种间进化分歧显示：东乡绿壳蛋鸡与同处江西省的泰和丝羽乌骨鸡遗传距离最近（0.009）；湖北省的郧县乌骨鸡与云南省的盐津乌骨鸡、腾冲雪鸡距离最远（0.019、0.020）。结合邻接系统发生树发现，国内10个乌鸡种群体共可归纳出6大分支起源，其中，与红色原鸡（Gallus gallus spadiceus）亚种同源的B分支包含的个体数最多（77/248，31.0%），A分支其次（69/248，27.8%）；东乡绿壳蛋鸡含3个分支，A分支最多（25/39，64.1%）。本研究表明，东乡绿壳蛋鸡的遗传多态性呈中等；同时，国内各地方乌鸡种之间存在较为明显的地域关联性，而其母系来源则各有偏重。
Abstract: Wugu chicken is a marvel of traditional Chinese poultry breeds and is characterized by black skin and bones, and grayish-black meat. There are nearly 17 species of Wugu chicken, not including breeds with partially black skin and bones, recorded in Animal Genetic Resources in China published by the China National Commission of Animal Genetic Resources. Among these breeds, the Dongxiang blue-eggshell chicken was originally bred in Dongxiang County, Jiangxi Province, China. This breed is characterized by black feathers and most of the crest, beak, skin, muscle, bone, shins, and its toes are also black. The unique characteristic of the Dongxiang chicken is a blue eggshell. Mitochondrial DNA (mtDNA) analysis has become an effective method to explore genetic relation among populations because of its low molecular mass, simple structure, rapid evolutionary rate, maternal inheritance, and non-tissue specification, among other features. The aim of the present study was to analyze mtDNA control region polymorphisms and evolution in conservative groups of Dongxiang blue-eggshell chickens in comparison to other domestic Wugu breeds to better elucidate the genetic structure of Dongxiang chickens to further develop conservation and exploitation programs.
Blood samples were collected from 39 (19 cocks and 20 hens) eleventh generation Dongxiang blue-eggshell conservative chickens and DNA was extracted and then PCR-amplified and the products were sequenced. Nine other samples from domestic Wugu chicken breeds bred by our team or collected from the NCBI database were used for nucleotide sequence analysis. All breeds used in this study were recorded in Animal Genetic Resources in China and all collected data were analyzed using DNASTAR 5.0, DNASP 5.10, and MEGA 5.10 softwares.
The results showed that the 534 bp mtDNA control region sequence of the Dongxiang blue-eggshell chicken had a G+C content of 43.1% and included 21 variable sites, of which 71.4% (15/21) were transitional and 28.6% (7/21) were transversional. The nucleotide diversity was 0.008 29 ± 0.001 30. There were 73 haplotypes found in 248 samples from 10 Wugu chicken breeds. The Dongxiang blue-eggshell chicken had nine haplotypes with a diversity of 0.773 ± 0.048. Estimates of evolutionary divergence between breeds showed that the Dongxiang blue-eggshell chicken was genetically closest (0.009) to the Taihe silky chicken, which was also originally bred in Jiangxi Province. Yunxian Wugu chicken originally bred in Hubei Province had the farthest genetic distance (0.019, 0.020) from Yanjin Wugu and Tengchong snow chickens originally bred in Yunnan Province. A neighbor-joining phylogeny tree revealed six original clades among 10 domestic Wugu chicken breeds. Clade B included Gallus gallus spadiceus  subspecies and had the largest number of samples (31.0%, 77/248), followed by clade A (27.8%, 69/248). The Dongxiang blue-eggshell chicken was closely related to three clades, of which clade A was the largest (64.1%, 25/39).
It is concluded that the Dongxiang blue-eggshell chicken is moderately genetically diverse and there is an apparent geographical association between domestic Wugu chicken breeds and their maternal origins, although each had individual characteristics.
出版日期: 2014-01-20
CLC:  S 831.2  
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王欢欢1
章学东1*
李庆海1
张成先2
楼立峰1
陈贤惠2

引用本文:

章学东1*， 李庆海1， 张成先2， 陈贤惠2， 楼立峰1， 王欢欢1. 东乡绿壳蛋鸡线粒体DNA控制区多态性及与9种地方乌鸡的进化分析[J]. 浙江大学学报(农业与生命科学版), 2014, 40(1): 103-110.

Zhang Xuedong1*, Li Qinghai1, Zhang Chengxian2, Chen Xianhui2, Lou Lifeng1, Wang Huanhuan1. Mitochondrial DNA control region polymorphisms in Dongxiang blue-eggshell chicken and their evolutionary relatedness with nine domestic Wugu breeds. Journal of Zhejiang University (Agriculture and Life Sciences), 2014, 40(1): 103-110.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2013.06.012    或    http://www.zjujournals.com/agr/CN/Y2014/V40/I1/103

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