四川三例疑似偶蹄目动物的分子鉴定

测定了3例来自四川崇州的疑似偶蹄目动物样本细胞色素b基因(Cytb),利用NCBI数据库中的BLAST功能对目标序列进行相似性搜索,发现与所测序列相似性高的分别为偶蹄目鹿科毛冠鹿Elaphodu scephalophus和牛科台湾鬣羚Capricornis swinhoei;联合下载四川有分布偶蹄目动物4科19种以及1条台湾鬣羚Cytb基因序列进行系统发育关系重建分析,结果显示有两例待鉴定样本(4#、5#)以置信度值100与毛冠鹿聚在一起;另一例待鉴定样本(1#)对应序列和鬣羚Capricornis milneedwardii聚在一起后与台湾鬣羚形成姊妹关系,然后以置信度值100和斑羚Naemorhedus griseus又形成姊妹关系。因此,3例疑似偶蹄目动物有2例可能为毛冠鹿,1例可能为鬣羚;在MEGA4.0软件中以Kimura2-parameter模型计算2例疑似为毛冠鹿(4#、5#)的目标序列与GenBank上毛冠鹿及赤麂Mun-tiacus muntjak、小麂Muntiacus reevesi序列间的遗传距离,发现两例疑似为毛冠鹿个体间遗传距离为0,2例疑似为毛冠鹿样本序列和下载的毛冠鹿序列间的遗传距离小(0.017),和赤麂、小麂间遗传距离大(0.119～0.120、0.116～0.119);1例疑似鬣羚(1#)的样本和下载鬣羚序列间遗传距离很小(0.006～0.009),和台湾鬣羚的遗传距离较大(0.043),和斑羚遗传距离更大(0.098)。结合四川有分布偶蹄目状况,认为3例待鉴定样本2例可能为毛冠鹿,1例可能是鬣羚。

Molecular Identification of Three Animals of Artiodactyla in Sichuan Province

We sequenced mitochondrial cytochrome b (cyt b) gene of 3 undetermined specimens of Artiodactyla, which were given from Chongzhou county, Sichuan province, China. The results of BLAST through NCBI website showed that one target sequence of the examined animal (1#) had great similarity to Taiwan serow and another two (4# and 5#) had great similarity to tufted deer. In order to build the phylogenetic tree, forty-four cyt b sequences were downloaded from GenBank. These sequences represented 20 species, 4 family, which all distributed in Sichuan province except Taiwan serow. The phylogenetic trees were constructed by using maximum likelihood method through RAxML BlackBox. The results show that one sample (1#) clustered with Chinese serow first and then clustered with Taiwan serow, finally formed a sister clade with Chinese goral. However, 4# and 5# clustered together with tufted deer with high bootstrap value (100). Therefore, 1# may be Chinese serow and 4# and 5# may be tufted deer. In addition, we calculated the genetic distance based on Kimura 2-parameter in MEGA 4.0. The result showed that the genetic distance between 1# and Chinese serow was very small (0.006~0.009), but that between 1# and Capricornis swinhoei and Naemorhedus griseus were large (0.043 and 0.098). The genetic distance between 4# and 5# was 0, and that between them and tufted deer was only 0.017, but that between them and Muntiacus muntjak and Muntiacus reevesi were 0.119~0.120 and 0.116~0.119. Thus, according to the current distribution of Artiodactyla in Sichuan provice and the phylogenetic tree and genetic distance, 1# should belong to Chinese serow and 4# and 5# to the tufted deer.