夜蛾颗粒体病毒宿主筛选及其增殖因子主次研究

Host Screening and Proliferation Factors of MOGV GXW9-3 Primary and Secondary Research

摘要: 夜蛾颗粒体病毒(GXW9-3 Mamestra oleracea granulosis virus)简称MOGV GXW9-3。该病毒2013年首次由高小文在江苏镇江发现，保藏于中国典型培养物保藏中心，保藏编号为CCTCCNO：V201340，样品呈黑色粉末状，保存于4℃冰箱。MOGV GXW9-3是寄生在鳞翅目夜蛾科昆虫真皮、脂肪组织及血细胞中的一种最重要的杆状病原病毒，以蛋白质包涵体的形式存在，蛋白质包含着一个病毒颗粒，核酸为双链DNA。本文研究影响MOGV GXW9-3增殖宿主及增殖因子的主次，筛选该病毒增殖最佳宿主及其增殖因子主次。研究结果表明，影响MOGV GXW9-3产量的因子及互作因子主次顺序为：宿主细胞 > 饲毒虫日龄与收毒时间互作 > 饲毒虫日龄与收毒时间互作 > 收毒时间 > 温度。明确饲毒宿主，饲毒虫日龄因子对病毒产量有极显著影响，饲毒虫日龄和饲毒虫日龄与收毒时间存在互作且对产量有显著影响。确定MOGV GXW9-3增殖宿主为斜纹夜蛾细胞，最佳生产条件为26~28℃、分别按药剂浓度为：6 × 10⁶ GV/mL、4 × 10⁶ GV/mL、3 × 10⁶ GV/mL、2.4 × 10⁶ GV/mL、2 × 10⁶ GV/mL的剂量饲毒斜纹夜蛾6龄幼虫，14~16 d后提取病毒，斜纹夜蛾6龄幼虫平均单虫个体重约1.046 g，冷冻干燥后每头虫体重量约为：0.085~0.108 g，平均单虫病毒产量约为0.069~0.28亿GV/g，幼虫孵化到收集幼虫提取病毒的生产周期为24 d，病毒增殖倍数为：31.25~43.75倍。

Abstract: GXW9-3 Mamestra oleracea granulosis virus is abbreviated as MOGV GXW9-3. The virus was first discovered by Xiaowen Gao in Zhenjiang, Jiangsu Province in 2013 and was stored in the Chinese Typical Culture Preservation Center with the storage number CCTCCNO: V201340. The samples were in black powder form and stored in a refrigerator at 4°C. MOGV GXW9-3 is one of the most im-portant rod pathogenic viruses in the dermis, adipose tissue and blood cells of lepidoptera. It exists in the form of protein inclusion body. The protein contains a virus particle and the nucleic acid is double-stranded DNA. In this paper, complex factors were used to study the primary and secondary order of MOGV GXW9-3 proliferation hosts and factors, and to screen the best host of the virus pro-liferation and its proliferation influence factors. The results showed that the main order of factors and interaction factors affecting the yield of MOGV GXW9-3 was as follows: host cell > interaction between day age of feeding insects and time of toxification > interaction between day age of feeding insects and time of toxification > time of toxification > temperature. Clearly defining the host of the virus, the virus yield was significantly influenced by the age factors of the feed worms. The age of the feed worms and the age of the feed worms interacted with the time of receiving the poison and had a significant effect on the yield. The proliferation host of MOGV GXW9-3 was determined to be moth cells, and the optimal production conditions were 26~28˚C, according to the drug concentration: The 6th instar larvae were fed with 6 × 10⁶ GV/mL, 4 × 10⁶ GV/mL, 3 × 10⁶ GV/mL, 2.4 × 10⁶ GV/mL, 2 × 10⁶ GV/mL, and the virus was extracted 14~16 days later. The average body weight of the 6th instar larvae was about 1.046 g. After freeze-drying, the weight of each insect was about 0.085~0.108 g, the average virus production was about 0.069~0.28 million GV/g, the production cycle from larvae hatching to larvae collection and virus extraction was 24 d, and the multiplication of virus was 31.25~43.75 times.