刘爽,王一凡,黄春华,刘德芳,张书环,楼迪栋
LIU Shuang,WANG Yifan,HUANG Chunhua,LIU Defang,ZHANG Shuhuan,LOU Didong

• 1:贵州中医药大学基础医学院
• 2:贵州中医药大学法医中药毒理学特色重点实验室
• 3:贵州中医药大学司法鉴定所

摘要(Abstract)：

目的 利用东北马鹿微卫星DNA遗传标记用于鹿药材的生物鉴别。方法 采用微卫星识别工具(MISA)对欧洲马鹿和中亚马鹿基因组进行生物信息学分析，获得东北马鹿同源微卫星DNA 20个遗传标记；随机选择2个具有一定多态性的微卫星遗传标记，对随机抽取的48头东北马鹿血样DNA进行PCR-STR扩增、电泳分离和分型、等位基因测序及群体遗传学参数调查。结果 CM41S16和CM12S18基因座具有高度多态性，群体遗传学参数显示CM41S16和CM12S18基因座均有5个等位基因及15种基因型，等位基因频率分布范围分别为0.041 7～0.333 3和0.072 9～0.333 3,多态性信息量(PIC)为0.620 6和0.721 2,个体识别率(DP)分别为0.839 6和0.903 5。结论 CM41S16和CM12S18具有高度多态性，可以用于鹿药材生物鉴别。
Objective To implement the biological identification of deer medicine by using micro-satellite DNA genetic markers of Northeast red deer(Cervus elaphus xanthopygus). Methods MIcro-SAtellite identification tool(MISA) was used to conduct bioinformatic analysis on the genomes of European and Central Asian red deer, and 20 genetic markers of homologous micro-satellite DNA of Cervus elaphus xanthopygus were obtained. Two microsatellite genetic markers with certain polymorphism were randomly selected to perform PCR-STR amplification, electrophoretic isolation and typing, allele sequencing and population genetic parameter investigation of the DNA from 48 randomly selected Cervus elaphus xanthopygus blood samples. Results CM41S16 and CM12S18 loci showed high gene polymorphism. In the population genetic parameters: CM41S16 and CM12S18 loci had 5 alleles and 15 genotypes, the allele frequency distribution ranges were 0.041 7-0.333 3 and 0.072 9-0.333 3, the polymorphism information contents(PIC) were 0.620 6 and 0.721 2, and the individual probabilities of discrimination power(DP) were 0.839 6 and 0.903 5, respectively. Conclusion CM41S16 and CM12S18 are highly polymorphic and can be used for the biological identification of deer medicine.

关键词(KeyWords)： 东北马鹿;鹿血晶;微卫星;群体遗传学;生物识别
Cervus elaphus xanthopygus;deer blood crystal body;microsatellite;population genetics;biological recognition

Abstract:

Keywords:

基金项目(Foundation): 贵州省教育厅自然科学研究项目(黔教合KY字[2021]004);; 贵阳市科技计划项目(筑科合同[2020]-18-4);; 贵州省科技计划项目(黔科合支撑[2020]4Y209)

作者(Author): 刘爽,王一凡,黄春华,刘德芳,张书环,楼迪栋
LIU Shuang,WANG Yifan,HUANG Chunhua,LIU Defang,ZHANG Shuhuan,LOU Didong

参考文献(References)：

• [1] 韩欢胜，郭喜明，徐馨，等.鹿血功能研究进展[J].特种经济动植物，2021,24(5):31-32.
• [2] 李顺才，杜利强，安瑞永.中国野生马鹿资源的保护与利用[J].草业科学，2008,(4):82-85.
• [3] 秦瑜，张明海.中国马鹿的研究现状及展望[J].野生动物，2009,30(2):100-104.
• [4] 袁媛，徐德平，吴盼盼，等.鹿血中甾体化合物的分离与结构鉴定[J].食品与生物技术学报，2014,33(6):667-671.
• [5] 田新民，张明海，张辉，等.黑龙江省完达山东部林区马鹿种群遗传多样性的微卫星分析[J].生态学杂志，2010,29(3):543-548.
• [6] 张辉，张明海，罗理扬，等.黑龙江省完达山东部林区马鹿微卫星位点的筛选[J].野生动物，2010,31(2):59-62.
• [7] 尹君，张明海，谢绪昌，等.STR在野生东北马鹿个体识别中的应用[J].野生动物，2007,28(3):42-44.
• [8] BA H,CAI Z,GAO H,et al.Chromosome-level genome assembly of Tarim red deer,Cervus elaphus yarkandensis[J].Sci Data,2020,7(1):1-8.
• [9] BANA N A,NYIRI A,NAGY J,et al.The red deer Cervus elaphus genome CerEla1.0:sequencing,annotating,genes,and chromosomes[J].Mol Genet Genomics,2018,293(3):665-684.
• [10]耿广耀，由玉岩，刘群秀.豚鹿SSR标记开发与遗传多样性分析[J].野生动物学报，2022,43(3):816-820.
• [11]SIM Z,MONDERMAN L,HILDEBRAND D,et al.Development and implementation of a STR based forensic typing system for moose (Alces alces)[J].Forensic Sci Int Genet,2021,53:102536.
• [12]张苏云，席继锋，哈孜，等.鹿微卫星遗传分析的研究现状[J].草食家畜，2005,26(2):30-33.
• [13]张沼，张蕊，李晓宇，等.内蒙古赛罕乌拉自然保护区东北马鹿种群遗传多样性与性别结构分析[J].兽类学报，2021,41(1):42-50.
• [14]吴琼，邢秀梅，杨福合.分子遗传标记在鹿科动物遗传多样性研究中的应用[J].特产研究，2007,118(4):69-73.
• [15]WALSH P S,METZGER D A,HIGUCHI R,et al.Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material[J].Bio Techniques,1991,10(4):506-513.
• [16]张苏云，钟林，贾斌，等.新疆塔里木马鹿遗传多样性的微卫星分析[J].畜牧与兽医，2006,56(11):17-20.
• [17]JIA B,WANG G,ZHENG J,et al.Development of novel EST microsatellite markers for genetic diversity analysis and correlation analysis of velvet antler growth characteristics in Sika deer[J].Hereditas,2020,157(1):1-14.
• [18]张振祥，陈艳霞，习向玉，等.应用微卫星标记分析山丹军马场甘肃马鹿遗传多样性[J].高原科学研究，2022,6(1):63-75.
• [19]李文斌，闻亮，郑维平，等.微卫星标记分析5种鹿类动物群体遗传多样性[J].野生动物学报，2010,31(5):227-231.
• [20]LIU H,JU Y,TAMATE H,et al.Phylogeography of sika deer (Cervus nippon) inferred from mitochondrial cytochrome-b gene and microsatellite DNA[J].Gene,2021,772:145375.
• [21]任战军，熊建杰，邓铸疆，等.白唇鹿的微卫星遗传多态性分析[J].西北农业学报，2011,20(5):6-9.
• [22]高惠，乔付杰，滕丽微，等.马鹿阿拉善亚种遗传多样性和群体结构[J].兽类学报，2020,40(5):458-466.
• [23]房瑞新，田雪琪，邹晨，等.马鹿的起源进化与遗传多样性研究进展[J].中国畜牧兽医，2021,48(12):4575-4587.
• [24]陈艳霞，张振祥，肖荣荣，等.甘肃马鹿(Cervus elaphus kansuensis)转录组SSR和SNP特征分析[J].基因组学与应用生物学，2021,40(Z3):2976-2987.
• [25]夏米西丁·阿不都热依木，艾斯卡尔·买买提，布左拉·吐尔逊，等.野生天山马鹿哈密种群微卫星遗传多样性[J].野生动物学报，2014,35(1):38-44.

文章评论(Comment)：