速生薄口螨休眠体的形态和分子特征鉴定
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摘要
目的 通过形态和分子特征鉴定速生薄口螨休眠体。方法 从采集到的生姜样本中分离出速生薄口螨休眠体,分别在光镜和扫描电镜下观察其外部形态和超微形态并进行拍照鉴定。同时提取基因组DNA对cox1和ITS基因进行测序和分析。结果 速生薄口螨休眠体足4对,背面躯体呈扁平状,表皮骨化,前足体呈三角状,足三表皮内突相连,形成一条拱线,分隔开胸板和腹板。足爪和前跗节发达,有刚毛附着,躯体末端吸盘板发达,板上有八个吸盘呈2-4-2分布。经PCR扩增得到cox1基因片段大小为487 bp,ITS全序列长度为1667 bp。结论 根据光镜和电镜下速生薄口螨休眠体的形态,结合cox1和ITS基因序列,为速生薄口螨休眠体的鉴定和生物学分类提供了依据,也可为控制速生薄口螨及其引起的过敏性疾病奠定基础。
Objective To identify Histiostoma feroniarum hypopus according to the morphological and molecular characteristics. Methods The Histiostoma feroniarum hypopus was isolated from the collected ginger samples, and its external morphology and ultrastructure were observed under light microscope and scanning electron microscope, and photographed for identification. Meanwhile, genomic DNA was extracted for sequencing and analysis of cox1 and ITS genes. Results The Histiostoma feroniarum hypopus was composed of 4 pairs of feet, the dorsal body was flat, the the epidermis was of significant ossification, the forefoot body was triangular, and the epidermal processes of foot III were connected to form an arch line separating the thoracic plate from the abdominal plate. The foot claw and anterior tarsus were developed, there were bristles attached, the sucker plate at the end of the body was developed, and there were eight suckers on the plate with 2-4-2 distribution. The size of cox1 gene fragment obtained by PCR amplification was 487 bp, and the total sequence length of ITS was 1667 bp. Conclusion The morphology of the Histiostoma feroniarum hypopus under light microscope and electron microscope combined with the cox1 and ITS gene sequences provide a basis for the identification and biological classification of the Histiostoma feroniarum hypopus and also lay a foundation for the control of the Histiostoma feroniarum hypopus and allergic diseases caused by it.
Objective To identify Histiostoma feroniarum hypopus according to the morphological and molecular characteristics. Methods The Histiostoma feroniarum hypopus was isolated from the collected ginger samples, and its external morphology and ultrastructure were observed under light microscope and scanning electron microscope, and photographed for identification. Meanwhile, genomic DNA was extracted for sequencing and analysis of cox1 and ITS genes. Results The Histiostoma feroniarum hypopus was composed of 4 pairs of feet, the dorsal body was flat, the the epidermis was of significant ossification, the forefoot body was triangular, and the epidermal processes of foot III were connected to form an arch line separating the thoracic plate from the abdominal plate. The foot claw and anterior tarsus were developed, there were bristles attached, the sucker plate at the end of the body was developed, and there were eight suckers on the plate with 2-4-2 distribution. The size of cox1 gene fragment obtained by PCR amplification was 487 bp, and the total sequence length of ITS was 1667 bp. Conclusion The morphology of the Histiostoma feroniarum hypopus under light microscope and electron microscope combined with the cox1 and ITS gene sequences provide a basis for the identification and biological classification of the Histiostoma feroniarum hypopus and also lay a foundation for the control of the Histiostoma feroniarum hypopus and allergic diseases caused by it.
引文
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[2] 莫颂轶,蒋敏丽,孔保庆,等.239名壮、汉族城乡女大学生面部蠕形螨感染情况及影响因素分析[J].右江民族医学院学报,2013,35(2):141-143.
[3] 李朝品,武前文.房舍和储藏物:粉螨[M].北京:中国科学技术出版社,1996:376-378.
[4] 李朝品,沈兆鹏.中国粉螨概论[M].北京:中国科学技术出版社,2016:454-462.
[5] 温硕洋,何晓芳.一种适用于昆虫痕量DNA模板制备的方法[J].昆虫知识,2003,40(3):276-279.
[6] 孙恩涛.椭圆食粉螨线粒体基因组测序及种群遗传结构的研究[D].芜湖:安徽师范大学,2014.
[7] 张曼丽,范青海.螨类休眠体的发育与治理[J].昆虫学报,2007,50(12):1293-1299.
[8] 赵晓平,刘晓光.实验室薄口螨的大量培养及休眠体的诱导[J].动物学杂志,2011,46(4):42-46.
[9] 段彬彬,湛孝东,宋红玉,等.食用菌速生薄口螨休眠体光镜下形态观察[J].中国血吸虫病防治杂志,2015,27(4):414.
[10] 洪勇,陶宁,湛孝东,等.洋葱害螨速生薄口螨的形态观察[J].中国血吸虫病防治杂志,2016,28(3):301-303.
[11] 柴强,郭娇娇,陶宁,等.中药材生姜孳生速生薄口螨的情况调查及形态观察[J].中国血吸虫病防治杂志,2017,29(6):776-777.
[12] 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,2018,41(2):1-11.
[13] 吴太葆,夏斌,邹志文,等.椭圆食粉螨线粒体DNACOⅠ基因片段序列分析[J].蛛形学报,2007,16(2):79-82.
[14] 孔里微.三种叶螨的线粒体基因组进化机制研究[D].南京:南京农业大学,2016.
[15] Ben DT,Melamed S,Gerson U,et al.ITS2 sequences as barcodes for identifying and analyzing spider mites (Acari:Tetranychidae)[J].Experimental and Applied Acarology,2007,41(3):169-181.
[16] 邹志文,陈芬,夏斌,等.几种钝绥螨ITS基因片段的序列分析[J].中国农业科学,2011,44(23):4945-4951.