目的 建立马六甲肉食螨与转开肉食螨的形态学和分子鉴定方法。方法 2018年5月至2019年7月，从安徽省部分地区农户粮仓的储藏物样本中发现并采集马六甲肉食螨和转开肉食螨，比较2种肉食螨的形态特征，使用MEGA X软件对2种肉食螨的线粒体细胞色素C氧化酶亚基Ⅰ（COⅠ）基因序列进行比对分析，最大似然法构建系统发育树。结果 马六甲肉食螨与转开肉食螨形态特征相似，但在光镜下仍可通过气门板形状、足Ⅰ跗节管毛的形态及其与支持毛的相对比例等特征进行形态学鉴定。基于COⅠ基因种间遗传距离>0.02，种内遗传距离均<0.02，碱基组成分析发现此段序列具有明显的A+T偏向性，BLAST比对结果与形态学鉴定一致，系统发育树显示转开肉食螨与马六甲肉食螨均能各自聚为一支。结论 马六甲肉食螨和转开肉食螨成螨可以依据外部形态特征进行形态学鉴定，也能基于线粒体COⅠ基因进行分子鉴定。2种鉴定方法相结合，可提高近缘种鉴别的准确性。
Objective To investigate methods for the morphological and molecular identification of Cheyletus malaccensis and C. carnifex. Methods From May 2018 to July 2019, C. malaccensis and C. carnifex were obtained from the storage samples of some farmers' granaries in several areas of Anhui province. The morphological characteristics of C. malaccensis and C. carnifex were compared and described. MEGA X software was used to analyze mitochondrial cytochrome c oxidase subunit Ⅰ (COⅠ) gene sequence for C. malaccensis and C. carnifex, and the maximum likelihood method was used to construct the phylogenetic tree. Results C. malaccensis and C. carnifex had similar morphological characteristics, while morphological identification could be performed under a light microscope based on the shape of peritremes, the morphology of guard seta of tarsus I, and the length-ratio between solenidia and pretarsus of tarsus I. Based on the COⅠ gene, the interspecific genetic distance was >0.02 and the intraspecific genetic distance was <0.02. The base composition analysis showed that the sequence had an obvious A+T bias, and the results of BLAST were consistent with those of morphological identification. Phylogenetic tree showed that C. malaccensis and C. carnifex were clustered into one single branch, respectively. Conclusion Identification of C. malaccensis and C. carnifex can be performed based on the external morphological characteristics and the mitochondrial COⅠ gene, and the combination of these two methods can improve the accuracy of the identification of closely related species.
[1] Bochkov AV. Mites of the family Cheyletidae (Acari:Prostigmata):phylogeny,distribution,evolution and analysis of parasite-host relationship[J]. Parazitologiia,2004,38(2):122-138.
[2] Xia B. A review of progress on the systematics and biology of the family Cheyletidae in China,with a checklist of the Chinese cheyletids[J]. Zoosymposia,2010,4(1):158-164. DOI:10. 11646/zoosymposia.4.1.11.
[3] Yang XQ,Ye QT,Xin TR,et al. Population genetic structure of Cheyletus malaccensis (Acari:Cheyletidae) in China based on mitochondrial COⅠand 12S rRNA genes[J]. Exp Appl Acarol,2016,69(2):117-128. DOI:10.1007/s10493-016-0028-3.
[4] 李朝品,湛孝东,孙恩涛,等. 储藏中药材孳生肉食螨种类及其群落生态研究[J]. 中药材,2013,36(9):1412-1416. DOI:10.13863/j.issn1001-4454.2013.09.004.Li CP,Zhan XD,Sun ET,et al. Investigation on species and community ecology of cheyletoid mites breeding in the stored traditional Chinese medicinal materials[J]. J Chin Med Mater,2013,36(9):1412-1416.DOI:10.13863/j.issn1001-4454.2013. 09.004.
[5] 陈静,王康,南丹阳,等. 芜湖市某高校学生宿舍床席螨类和昆虫孳生状况调查[J]. 中国血吸虫病防治杂志,2016,28(2):151-155. DOI:10.16250/j.32.1374.2015231.Chen J,Wang K,Nan DY,et al. Investigation on mites and insects breeding status of bed mats in college dormitories in Wuhu city[J]. Chin J Schistosom Control,2016,28(2):151-155. DOI:10.16250/j.32.1374.2015231.
[6] Beroiz B,Couso-Ferrer F,Ortego F,et al. Mite species identification in the production of allergenic extracts for clinical use and in environmental samples by ribosomal DNA amplification[J]. Med Vet Entomol,2014,28(3):287-296. DOI:10.1111/mve.12052.
[7] 李朝品. 医学节肢动物学[M]. 北京:人民卫生出版社,2009:1266-1272.Li CP. Medical arthropodology[M]. Beijing:People's Medical Publishing House,2009:1266-1272.
[8] Corpuz-Raros LA. Twelve new species and one new record of Cheyletidae (Acari) from the Philippines[J]. Int J Acarol,1998,24(4):259-290. DOI:10.1080/01647959808683594.
[9] 易忠权,夏伟,赵盼雯,等. DNA标记技术在螨类系统学研究中的应用[J]. 现代生物医学进展,2017,17(31):6196-6200. DOI:10.13241/j.cnki.pmb.2017.31.046.Yi ZQ,Xia W,Zhao PW,et al. DNA markers in the systematic study of mites[J]. Progr Mod Biomed,2017,17(31):6196-6200. DOI:10.13241/j.cnki.pmb.2017.31.046.
[10] 邓洁,郭浩杰,黄少康,等. 基于线粒体COⅠ基因的印度新曲厉螨的分子鉴定[J]. 环境昆虫学报,2019,41(2):380-386. DOI:10.3969/j.issn.1674-0858.2019.02.21.Deng J,Guo HJ,Huang SK,et al. Molecular identification of Neocypholaelaps indica Evans based on mitochondrial COⅠgene[J]. J Environ Entomol,2019,41(2):380-386. DOI:10.3969/j.issn.1674-0858.2019.02.21.
[11] Li MN,Yang QQ,Shao HF,et al. Species identification and phylogenetic relationships among five species of psocids from Chinese herbal medicines[J]. J Stored Prod Res,2020,85:101539. DOI:10.1016/j.jspr.2019.101539.
[12] Ernieenor FCL,Ernna G,Jafson AS,et al. PCR identification and phylogenetic analysis of the medically important dust mite Suidasia medanensis (Acari:Suidasiidae) in Malaysia[J]. Exp Appl Acarol,2018,76(1):99-107. DOI:10.1007/s10493-018-0285-4.
[13] Sakamoto H,Matsuda T,Suzuki R,et al. Molecular identification of seven species of the genus Stigmaeopsis (Acari:Tetranychidae) and preliminary attempts to establish their phylogenetic relationship[J]. Syst Appl Acarol,2017,22(1):91-101. DOI:10.11158/saa.22.1.10.
[14] Webster LMI,Thomas RH,McCormack GP. Molecular systematics of Acarus siro s. lat.,a complex of stored food pests[J]. Mol Phylogenet Evol,2004,32(3):817-822. DOI:10.1016/j.ympev.2004.04.005.
[15] 郑凌霄,尹灿灿,王逸枭,等. 基于DNA条形码技术的粉螨种类鉴定研究[J]. 中国媒介生物学及控制杂志,2019,30(2):180-184. DOI:10.11853/j.issn.1003.8280.2019.02.015.Zheng LX,Yin CC,Wang YX,et al. Identification of acaroid mite species based on DNA barcoding[J]. Chin J Vector Biol Control,2019,30(2):180-184. DOI:10.11853/j.issn.1003.8280.2019. 02.015.
[16] Yang B,Cai JL,Cheng XJ. Identification of astigmatid mites using ITS2 and COⅠregions[J]. Parasitol Res,2011,108(2):497-503. DOI:10.1007/s00436-010-2153-y.
[17] Palasio RGS,de Almeida Guimarães MC,Ohlweiler FP,et al. Molecular and morphological identification of Biomphalaria species from the state of São Paulo,Brazil[J]. ZooKeys,2017(668):11-32. DOI:10.3897/zookeys.668.10562.
[18] Hebert PDN,Cywinska A,Ball SL,et al. Biological identifications through DNA barcodes[J]. Proc Roy Soc B:Biol Sci,2003,270(1512):313-321. DOI:10.1098/rspb.2002.2218.
