基于Cytb基因的穿山甲科物种鉴定研究王辰1,杨娅富1,李立2,逯金瑶2,李嘉昊1,李波1,3∗(1.东北林业大学野生动物与自然保护地学院,黑龙江哈尔滨150040;2.湖南省生物多样性保护中心,湖南长沙410000;3.国家林业和草原局野生动植物检测中心,黑龙江哈尔滨150040)摘要为鉴别混有非穿山甲物种疑似检材的来源物种,基于细胞色素b基因设计了穿山甲科特异的新引物DCW-F1/DCW-R1。试验表明:其特异性良好、灵敏度较高、适用检材类型较广。采用BLAST比对法、遗传距离法和系统发育法分析了4组不同长度Cytb序列,研究序列长度对穿山甲物种鉴定的影响。结果表明,多数样本与相应穿山甲物种同源序列相似度均高于98%,建议将该值作为BLAST比对法的判别标准;除马来穿山甲(211bp)和树穿山甲(421和211bp)外,剩余5个穿山甲物种种间最小遗传距离均大于种内最大遗传距离,序列长度的变化和样本量的不平衡会导致该比值不稳定;除马来穿山甲和菲律宾穿山甲,其余物种均为单系,序列长度变短使得系统发育树拓扑结构和分支置信度发生相应变化。关键词物种鉴定;穿山甲;细胞色素b基因(Cytb);序列分析中图分类号S863文献标识码A文章编号0517-6611(2023)03-0089-07doi:10.3969/j.issn.0517-6611.2023.03.020开放科学(资源服务)标识码(OSID):SpeciesIdentificationforPangolinsBasedonCytochromebGeneWANGChen1,YANGYa-fu1,LILi2etal(1.CollegeofWildlifeandNaturalReserve,NortheastForestryUniversity,Harbin,Hei-longjiang150040;2.HunanBiodiversityConservationCenter,Changsha,Hunan410000)AbstractApairofnewspecificprimersDCW-F1/DCW-R1weredesignedbasedonCytochromeb(Cytb)genetoidentifysuspectedspeci-mensmixedwithnon-pangolinspecies.Theresultsshowedthatitwasgoodspecificity,highsensitivityandcouldbeusedforseveraltypesam-ples.Secondly,weusedBLASTcomparison,geneticdistanceandconstructingphylogenetictreetoanalyzethefourgroupsofCytbsequenceswithdifferentlengths,inordertoestimatetheinfluenceofchangingsequencelengthonpangolinidentification.Theresultsindicatedthatthesimilarityofhomologoussequencebetweenmostsamplesandcorrespondingpangolinwashigherthan98%.Thissuggestedthatthevalueshouldbeusedasthecriteriontoidentifypangolinspecies.Theminimuminter-specificgeneticdistance(inter-d)wasgreaterthanthemaximumIn-tra-specificgeneticdistan...
