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昆虫杆状病毒是一类寄生于节肢动物的囊膜包裹的双链环状DNA,杆状病毒作为新型生物杀虫剂逐渐应用于害虫防治。BmK IT是一种从东亚钳蝎分离出来的昆虫毒素,可以与昆虫细胞膜的钠离子通道相互作用,引起钠离子通道动作电位重复发放,并增强钠电流峰值,延缓钠传导关闭,使昆虫产生兴奋性麻痹。为了提高杆状病毒苜蓿银纹夜蛾核型多角体病毒(Autographa californica multicapsid nucleopolyhedrovirus,AcMNPV)杀虫活性,本研究将BmK IT基因插入到AcMNPV基因组中,形成重组杆状病毒AcMNPV-BmK IT。采用MTT法、TUNEL试剂、Western blot、real-time PCR及病毒滴度测定来检测AcMNPVBmK IT对草地夜蛾卵巢细胞Sf9发生凋亡及病毒在Sf9细胞内的复制情况。结果显示AcMNPV介导BmK IT的表达促进了Sf9细胞凋亡及病毒复制。此结果为揭示重组病毒AcMNPV-BmK IT的抗虫机制提供了实验依据。
Baculovirus is a type of double-stranded circular DNA encapsulated by arthropods. As a new biopesticide, baculovirus is gradually applied to pest control. BmK IT is an insect toxin isolated from Scorpion in East Asia that interacts with the sodium ion channels of insect cell membranes, causing repeated release of sodium ion channel action potentials and increasing peak sodium currents, slowing sodium conduction off, and exiting the insects Paralysis. In order to improve the insecticidal activity of the baculovirus Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV), the BmK IT gene was inserted into the AcMNPV genome to form the recombinant baculovirus AcMNPV-BmK IT. MTT assay, TUNEL reagent, Western blot, real-time PCR and virus titer assay were used to detect the apoptosis of Sf9 cells and the replication of Sf9 cells in AcMNPVBmK IT. The results showed that AcMNPV-mediated BmK IT expression promoted Sf9 cell apoptosis and viral replication. The results provide an experimental basis for revealing the insect-resistant mechanism of the recombinant virus AcMNPV-BmK IT.