球孢白僵菌NJBb2101发酵液对Sf9细胞的增殖抑制和凋亡诱导作用  

The effects of the fermentation filtrate from Beauveria bassiana NJBb2101 on the cell viability and apoptosis of Sf9 cells

作　　者：胡媛媛 王照英 张懿熙 刘泽文[1] HU Yuanyuan;WANG Zhaoying;ZHANG Yixi;LIU Zewen(College of Plant Protection,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学植物保护学院,江苏南京210095

出　　处：《南京农业大学学报》2020年第3期446-451,共6页Journal of Nanjing Agricultural University

基　　金：国家重点研发计划项目(2019YFD0300103)。

摘　　要：[目的]本文分析了对多种昆虫具有高毒力的球孢白僵菌菌株NJBb2101发酵液产物对细胞增殖抑制和凋亡诱导的作用,旨在为筛选球孢白僵菌杀虫成分提供理论基础。[方法]以昆虫细胞Sf9为材料,利用Cell Counting Kit-8(CCK-8)试剂盒检测发酵液产物对细胞增殖的抑制作用;利用AO/EB(吖啶橙-溴化乙锭)染色和Annexin V-FITC/PI(Annexin V-fluorescein isothiocyante/propidium iodide)染色-流式细胞仪2种检测方法测定了高、低质量浓度(1.92和0.48 mg·mL^-1)发酵液产物对细胞凋亡的诱导作用。[结果]NJBb2101发酵液产物对Sf9细胞增殖具有很强的抑制作用,测定的抑制中浓度(IC50)为0.907 mg·mL^-1(95%置信限为0.500~3.075 mg·mL^-1)。AO/EB和Annexin V-FITC/PI染色-流式细胞仪检测发现高浓度(1.92 mg·mL^-1)发酵液产物可以显著增强细胞凋亡率,平均凋亡率均在70%以上。[结论]球孢白僵菌发酵液中存在有效的杀虫成分,对昆虫细胞表现出增殖抑制和诱导凋亡的作用。[Objectives]An insecticidal strain of Beauveria bassiana NJBb2101 showed high toxic to several insect species.Here we want to evaluate the effects of the fermentation filtrate from B.bassiana on the cell viability and apoptosis of Sf9(Spodoptera frugiperda ovary cells).[Methods]The inhibition effects of the fermentation filtrate on Sf9 cells were tested using Cell Counting Kit-8(CCK-8).The effects of the fermentation filtrate on Sf9 apoptosis were determined through AO/EB(acridine orange/ethidium bromide)and Annexin V-FITC/PI(Annexin V-fluorescein isothiocyante/propidium iodide)method.[Results]The fermentation filtrate showed high inhibition effects on Sf9 viability with IC50 of 0.907 mg·mL^-1(95%confidence interval was 0.500-3.075 mg·mL^-1).Both AO/EB and Annexin V-FITC/PI methods showed that the high dose(1.92 mg·mL^-1)could significantly increase the apoptosis,with the average rate above 70%.[Conclusions]The results showed that some important insecticidal components existed in the the fermentation filtrate from B.bassiana,which affected Sf9 cells through cell viability and apoptosis aspects.

关 键 词：球孢白僵菌 NJBb2101 增殖抑制 细胞凋亡 

分 类 号：S476.12[农业科学—农业昆虫与害虫防治]