Chen C J, Quentin M E, Brennan L A, Kukel C, Thiem S M
Program in Genetics, Michigan State University, East Lansing 48224-1115, USA.
J Virol. 1998 Mar;72(3):2526-31. doi: 10.1128/JVI.72.3.2526-2531.1998.
The gypsy moth (Lymantria dispar) is nonpermissive for Autographa californica nucleopolyhedrovirus (AcNPV) infection. We previously isolated a gene, host range factor 1 (hrf-1), from L. dispar nucleopolyhedrovirus that promotes AcNPV replication in Ld652Y cells, a nonpermissive L. dispar cell line (S. M. Thiem, X. Du, M. E. Quentin, and M. M. Berner, J. Virol. 70:2221-2229, 1996). In the present study, we investigated the ability of hrf-1 to alter the larval host range of AcNPV. Bioassays using recombinant AcNPV bearing hrf-1 were conducted with insect larvae by use of oral infection. AcNPV bearing hrf-1 was infectious for neonate L. dispar larvae, with a 50% lethal concentration of 1.2 x 10(5) polyhedral inclusion bodies/ml of diet, which is similar to that of wild-type AcNPV for permissive hosts. AcNPV can kill neonate L. dispar larvae at high doses, but it does not kill third-instar larvae. However, electron microscopy studies of AcNPV-inoculated third-instar larvae revealed virus replication in the midgut cells. PCR analyses indicated that the virus was AcNPV. These results suggest that the block for AcNPV infection of L. dispar larvae is its inability to spread systematically from primary infection sites in the midgut epithelium and that this barrier is leaky in neonates. hrf-1 allows AcNPV to overcome this barrier. AcNPV recombinants bearing hrf-1 were also significantly more infectious for Helicoverpa zea, a resistant species, suggesting that the blocks for AcNPV infection of L. dispar and H. zea larvae may be similar.
舞毒蛾(Lymantria dispar)对苜蓿银纹夜蛾核型多角体病毒(AcNPV)感染不敏感。我们之前从舞毒蛾核型多角体病毒中分离出一个基因,宿主范围因子1(hrf - 1),它能促进AcNPV在Ld652Y细胞(一种对AcNPV不敏感的舞毒蛾细胞系)中复制(S. M. 蒂姆、X. 杜、M. E. 昆廷和M. M. 伯纳,《病毒学杂志》70:2221 - 2229,1996年)。在本研究中,我们研究了hrf - 1改变AcNPV幼虫宿主范围的能力。使用携带hrf - 1的重组AcNPV对昆虫幼虫进行口服感染生物测定。携带hrf - 1的AcNPV对初孵舞毒蛾幼虫具有感染性,其50%致死浓度为每毫升饲料1.2×10⁵个多角体包涵体,这与野生型AcNPV对敏感宿主的致死浓度相似。AcNPV在高剂量时能杀死初孵舞毒蛾幼虫,但不能杀死三龄幼虫。然而,对感染AcNPV的三龄幼虫进行电子显微镜研究发现病毒在中肠细胞中复制。PCR分析表明该病毒为AcNPV。这些结果表明,AcNPV感染舞毒蛾幼虫的障碍在于其无法从中肠上皮的初始感染部位系统性扩散,并且这种屏障在初孵幼虫中存在漏洞。hrf - 1使AcNPV能够克服这一屏障。携带hrf - 1的AcNPV重组体对抗性物种玉米夜蛾(Helicoverpa zea)的感染性也显著更高,这表明AcNPV感染舞毒蛾和玉米夜蛾幼虫的障碍可能相似。