Application of DNA barcoding of COI gene in identification of ticks in Fujian Province, China
ZHOU Shu-heng1, XIAO Fang-zhen1, LIU Wei-jun1, LIU Jing1, HAN Teng-wei1, DENG Yan-qin1, 2
1.Fujian Center for Disease Control and Prevention, Fujian Provincial Key Laboratory of Zoonosis Research,Fuzhou 350001,China; 2.College of Public Health, Fujian Medical University, Fuzhou 350108, China
Abstract：To explore the feasibility of mitochondrial cytochrome C oxidase subunit I(COI)gene based DNA barcoding for the identification of ticks in Fujian Province, China. Genomic DNA was extracted from 97 tick specimens belonging to 12 species of 5 genera by morphological identification. COI gene fragments were amplified by PCR, sequenced and aligned. The intra- and inter-species genetic distances were calculated by Kimura-2-parameter model, and a phylogenetic tree was constructed by the neighbor-joining (NJ) method.Results showed that the inter-species genetic distance (14.65%-26.79%) was significantly greater than the intra-species genetic distance (0-7.14%);BLAST homology comparison suggested that there was no COI gene sequence of Haemophysalis Yeni and Ixodes spinicoxalis in Genbank database. Morphological identification of H.flava, H.formosensis, H.hystricis, I.granulatus, Rhipicephalus microplus and R.sanguineus were consistent, while Amblyomma testudinarium, Dermacentor taiwanensis, I.sinensis were inconsistent with DNA barcode identification results. The NJ tree showed that different individuals from the same species formed monophyletic groups with high support value,and inter-species branches were clear.The study has shown that DNA barcoding determination of COI gene as new technique for tick identification is fast and accurate, but at this stage, DNA barcoding of COI gene can not be completely independent from traditional morphological identification, and they should be combined organically.
周淑姮, 肖方震, 刘维俊, 刘菁, 韩腾伟, 邓艳琴. COI基因DNA条形码技术在福建省蜱类鉴定中的应用[J]. 中国人兽共患病学报, 2020, 36(1): 25-31.
ZHOU Shu-heng, XIAO Fang-zhen, LIU Wei-jun, LIU Jing, HAN Teng-wei, DENG Yan-qin. Application of DNA barcoding of COI gene in identification of ticks in Fujian Province, China. Chinese Journal of Zoonoses, 2020, 36(1): 25-31.
[1] Horak IG, Camicas JL, Keirans JE. The Argasidae, Ixodidae and Nuttalliellidae (Acari: Ixodida) of the world: a list of valid species names[J]. Exp Appl Acarol,2002,28(1-4):27-54. DOI:10.1023/A:1025381712339 [2] 杨晓军,陈泽,刘敬泽.中国蜱类的有效属和有效种[J].河北师范大学学报(自然科学版),2008, 32(4):529-533. [3] 马广鹏, 孙传范, 赵娜, 等. 中国蜱传病主要流行趋势及防控科技对策[J].中国农业科技导报, 2011, 13(2):105-109. [4] 张瑞玲,王晓旭,张忠. DNA条形码技术在医学媒介生物鉴定中的应用[J].中国热带医学,2017,17(8):831-837. [5] Hebert PDN,Ratnasingham S,deWaard JR. Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species[J]. Proc Biol Sci, 2003, 270(1): 96-99. DOI:10.1098/rsbl.2003.0025 [6] Leo SST,Pybus MJ,Sperling FAH.Deep mitochondrial DNA lineage divergences within alberta populations of Dermacentor albipictus (Acari: Ixodidae) do not indicate distinct species[J]. J Med Entomol,2010,47(4):565-574. DOI:10.1603/ME10006 [7] Lv JZ,Wu SQ,Zhang YN, et al.Assessment of four DNA fragments (COI, 16S rDNA, ITS2, 12S rDNA) for species identification of the Ixodida (Acari: Ixodida)[J]. Parasit Vectors,2014,7(93):1-11. DOI:10.1186/1756-3305-7-93 [8] Zhang RL, Zhang B.Prospects of using DNA barcoding for species identification and evaluation of the accuracy of sequence databases for ticks (Acari: Ixodida)[J]. Ticks Tick Borne Dis, 2014, 5(3):352-358. DOI: 10.1016/j.ttbdis.2014.01.001 [9] 侯海燕. COX I基因DNA条形码技术在蜱类鉴别中的应用[J].国际检验医学杂志,2015,36(24):3570-3571. [10] Zhang R, Zhao A,Wang X,et al.Diversity of tick species on domestic animals in Shandong Province, China, using DNA barcoding[J]. Exp Appl Acarol,2017,73(1): 79-89. DOI:10.1007/s10493-017-0161-7 [11] Ondrejicka DA, Morey KC, Hanner RH.DNA barcodes identify medically important tick species in Canada[J]. Genome,2017,60(1):74-84. DOI:10.1139/gen-2015-0179 [12] Kwaka ML,Griffithsb J,Barry D, et al.The first DNA barcodes for the Australian platypus tick Ixodes ornithorhynchi Lucas, 1846 (Acari: Ixodidae) to facilitate conservation efforts for a declining parasite and its host[J]. Acarologia,2018,58(4):845-849. DOI:10.24349/acarologia/20184287 [13] Li LH, Zhang Y, Wang JZ, et al.High genetic diversity in hard ticks from a China-Myanmar border county[J]. Parasit Vectors,2018,11(469):1-11. DOI:10.1186/s13071-018-3048-5 [14] 邓国藩,姜在阶. 中国经济昆虫志,第39册,蜱螨亚纲,硬蜱科[M]. 北京:科学出版社,1991:1-359. [15] Wassef HY, Hoogstraal H.Dermacentor (Indocentor) auratus (Acari: Ixodoidea: Ixodidae): identity of male and female[J]. J Med Entomol,1984,21(2):169-173. [16] Apanaskevich MA,Apanaskevich DA.Reinstatement of Dermacentor bellulus (Acari: Ixodidae) as a valid species previously confused with D.taiwanensis and comparison of all parasitic stages[J]. J Med Entomol,2015,52(4):573-595. DOI:10.1093/jme/tjv034 [17] Hebert PDN,Cywinska A,Ball SL,et a1.Biological identiflcations through DNA barcodes[J]. P Roy Soc B-Biol Sci,2003,270(1512):313-321.DOI:10.1098/rspb.2002.2218 [18] 武伟,刘洪斌,张泽,等. 节肢动物线粒体基因组碱基组成特征分析[J]. 生物信息学,2007,3(5):102-105. [19] 吴荣泉,张建庆,方义亮,等.DNA条形码技术在福建省蝇类鉴定中的应用[J].中国媒介生物学及控制杂志,2014,12(25):514-517. [20] Bickford D, Lohman DJ, Sodhi NS, et al.Cryptic species as a window on diversity and conservation[J]. Trends Ecol Evol,2007,22(3): 148-155. DOI:10.1016/j.tree.2006.11.004 [21] Guglielmone AA, Robbins RG,Apanaskevich DA, et al.Comments on controversial tick (Acari: Ixodida) species names and species described or resurrected from 2003 to 2008[J]. Exp Appl Acarol, 2009,48(4): 311-327. [22] 周淑姮,肖方震,刘菁,等. 福建省印支革蜱亚属Subgenus Indocentor的分类研究[J].海峡预防医学杂志,2017,23(4):4-6. [23] 周淑姮,陈阳,林代华,等.福建省蜱类(蜱螨亚纲:蜱目)名录[J].寄生虫与医学昆虫学报,2016,23(4):230-236. [24] Yamaguti N, Tipton VJ, Keegan HL, et al.Ticks of Japan, Korea, and the Ryukyu islands[J]. Brigham Young University Science Bulletin,1971:1-226. [25] 陈念,付晓燕,赵树进,等. DNA条形码:物种分类和鉴定技术[J].生物技术通讯,2008,19(4):629-631. [26] 马英,鲁亮. DNA条形码技术[J].中国媒介生物学及控制杂志,2012,23(3):185-205.
