范张食酪螨和腐食酪螨的形态学与分子生物学鉴定  

Morphological and molecular biological identification of Tyrophagus fanetzhangorum and T.putrescentiae

作　　者：范笑忱 刘璐瑶 方瑜 褚凌渺 冯蕊 李飞燕 潘若兮 王少圣[1] 孙恩涛[1] FAN Xiao-chen;LIU Lu-yao;FANG Yu;CHU Ling-miao;FENG Rui;LI Fei-yan;PAN Ruo-xi;WANG Shao-sheng;SUN En-tao(Department of Hygiene Inspection and Quarantine,Wannan Medical College,Wuhu,Anhui 241002,China)

机构地区：[1]皖南医学院卫生检验与检疫学教研室,安徽芜湖241002

出　　处：《中国媒介生物学及控制杂志》2022年第2期277-280,共4页Chinese Journal of Vector Biology and Control

基　　金：国家自然科学基金(31870352);皖南医学院2019年青年卓越人才资助项目(wyqnyx201902);安徽省大学生创新创业教育训练计划项目(S202010368034)。

摘　　要：目的对范张食酪螨与腐食酪螨进行形态学和分子生物学鉴定。方法采集房舍和储藏物样本,从样本中挑取单个成螨,进行纯培养。从培养标本中挑取成螨制成玻片标本。根据无气门亚目螨类检索表,进行形态学鉴定。然后,提取单只螨基因组DNA,通过PCR扩增、测序获得线粒体细胞色素C氧化酶亚基Ⅰ基因(COⅠ)序列;将所获序列在美国国立生物技术信息中心中用BLAST进行同源性比对。使用ClustalⅩ1.83软件对2种螨的COⅠ序列进行比对,基于MEGAⅩ软件进行序列分析,并以最大似然法构建系统发育树。结果范张食酪螨足Ⅱ基节板后缘1/3处明显向内凹陷,下缘曲线近似“S”形,足Ⅰ跗节感棒ω1在远端2/3处明显增宽;而腐食酪螨足Ⅱ基节板呈三角形,下缘平直或略凹,足Ⅰ跗节感棒ω1在远端1/4处增宽。所测得的范张食酪螨和腐食酪螨COⅠ基因序列经BLAST比对显示,分别与GenBank中收录的范张食酪螨和腐食酪螨COⅠ基因序列相似性在99.00%以上。基于COⅠ基因构建的最大似然法系统进化树显示,范张食酪螨和腐食酪螨均独立聚为一支,且支持率均达100%。结论范张食酪螨和腐食酪螨既可以利用外部特征进行形态学鉴定,也可以基于COⅠ基因进行分子生物学鉴定;形态学和分子生物学鉴定方法相结合,可以提高螨类近缘种鉴定的准确性。Objective To identify Tyrophagus fanetzhangorum and T.putrescentiae by morphological and molecular methods.Methods Samples were collected from houses and stored products,and individual mites were selected for pure culture.The cultured adult mites were used to prepare slide specimens for morphological identification based on the taxonomic keys of Astigmata.Subsequently,genomic DNA was extracted from individual mites for the PCR amplification,and sequencing of mitochondrial cytochrome c oxidase subunitⅠgene(COⅠ).The resulting sequences were subjected to homology search using BLAST in NCBI.The COⅠsequences of T.fanetzhangorum and T.putrescentiae were aligned using ClustalⅩ1.83.Sequence analysis was performed using MEGAⅩ,and a phylogenetic tree was constructed using the maximum likelihood method.Results The posterior 1/3 of the coxal plateⅡof T.fanetzhangorum was concave,the inferior margin was"S"shaped,and the distal 2/3 of solenidionω1on tarsusⅠobviously widened.The coxal plateⅡof T.putrescentiae was triangular,the inferior margin was straight or slightly concave,and the solenidionω1on tarsusⅠwidened at the distal 1/4.The COⅠgene sequences of the isolated T.fanetzhangorum and T.putrescentiae showed>99.00%similarity with the COⅠgene sequences of T.fanetzhangorum and T.putrescentiae in GenBank.The phylogenetic tree showed that T.fanetzhangorum and T.putrescentiae clustered into a branch with a bootstrap value of 100%.Conclusion Identification of T.fanetzhangorum and T.putrescentiae can be performed based on morphological characteristics or molecular analysis of the COⅠgene.The combination of morphological and molecular identification can improve the accuracy of the identification of closely related mite species.

关 键 词：范张食酪螨 腐食酪螨 形态学鉴定 分子生物学鉴定 细胞色素C氧化酶亚基Ⅰ基因 

分 类 号：R384.4[医药卫生—医学寄生虫学]