四环素降低致倦库蚊共生Wolbachia密度方法的建立及效果评价

ESTABLISHMENT AND EFFECT EVALUATION OF A METHOD FOR REDUCING THE SYMBIOTIC WOLBACHIA DENSITY OF CULEX QUINQUEFASCIATUS WITH TETRACYCLINE

胞内共生菌Wolbachia在昆虫生理生殖和天然免疫方面发挥着重要作用,蚊虫体内的Wolbachia密度与其抗病毒作用有很强的相关关系。本研究建立了一种利用四环素处理幼虫降低致倦库蚊共生Wolbachia密度的方法,并采用实时荧光定量PCR进行了效果评估。结果显示,使用0.05 mg/mL四环素处理致倦库蚊海口株和温州株,处理后Wolbachia密度中位数分别下降2729和3507倍;卵巢及其余组织的Wolbachia密度均下降至0.001以下,其中,卵巢中Wolbachia密度下降825629倍。可见,0.05 mg/mL四环素处理幼虫能显著降低蚊虫体内Wolbachia密度,为后续清除Wolbachia、探究Wolbachia对致倦库蚊感染和传播病毒的影响提供了技术基础。

The intracellular symbiotic Wolbachia plays an important role in insect physiological reproduction and innate immunity,and its density in mosquitoes was shown strongly related the antiviral effect of mosquitoes.In this study,a method for reducing the symbiotic Wolbachia loads of Culex pipiens quinquefasciatus by tetracycline treatment on mosquito larvae was established,and effects was evaluated by real-time quantification PCR.The results showed that treated with optimal concentration tetracycline at 0.5 mg/mL,the median density of Wolbachia in Haikou,and Wenzhou strains decreased by 2729 times and 3507 times respectively.The Wolbachia loads in the ovaries and rest carcass decreased to less than 0.001,and the load in the ovaries decreased by 825629 times.Tetracycline treatment on larvae can significantly reduce the density of Wolbachia in Culex pipiens quinquefasciatus,which provides a reasonable strategy to explore the physiology roles of Wolbachia in maintaining and transmitting of mosquito borne pathogens in C.pipiens quinquefasciatus through eliminating or loads controlling of Wolbachia in host mosquitoes.