Biological Control of Insects Research Laboratory, USDA-ARS 1503 S. Providence Road, Research Park, Columbia, Missouri, 65203-3535 USA.
J Insect Sci. 2008;8:1-19. doi: 10.1673/031.008.4201.
A multiple nucleopolyhedrovirus (MNPV) was isolated from Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) larvae that had been stung by the parasitoid Cotesia marginiventris (Cresson) (Hymenoptera: Braconidae). The wild type virus was plaque purified by infecting a Heliothis subflexa (BCIRL- HsAM1) cell line and isolating several clones. The mean estimated genomic size of this virus based on PstI, BstEII, StyI, HindIII restriction profiles was estimated to be 106 +/- 2.5 kbp (mean+/-SE). A clone designated as TnMNPV/CmBCL9 was used in bioassays against several lepidopteran pests and in comparative studies with the baculoviruses AcMNPV, AgMNPV, AfMNPV, PxMNPV and HzSNPV of Autographa califomica, Anticarsia gemmatalis, Anagrapha falcifera, Plutella xylostella, and Helicoverpa zea, respectively. Infectivity studies showed that TnMNPV/CmBCL9 was highly infectious for Heliothis subflexa and T. ni, with an LC(50) value 0.07 occlusion bodies/mm(2) in both species and also infectious for H. zea and Heliothis virescens with LC(50) values of 0.22 and 0.27 occlusion bodies/mm(2), respectively. Restriction endonuclease analysis of the isolate and selected baculoviruses revealed profiles that were very similar to AfMNPV but different from the restriction endonuclease profiles of the other baculoviruses. Hybridization studies suggest that the TnMNPV/CmBCL9 was closely related to AfMNPV and AcMNPV-HPP. Further support for this comes from a phylogenetic analysis employing a split-graphs network, comparing the polh, egt, and p10 genes from TnMNPV/CmBCL9 with those from other baculoviruses and suggests that this virus is closely related to the AcMNPV variants, AfMNPV and RoMNPV of Rachiplusia ou.
一种多粒包埋型核型多角体病毒(MNPV)从被寄生蜂 Cotesia marginiventris(膜翅目:Braconidae)刺伤的烟夜蛾(Trichoplusia ni)幼虫中分离出来。野生型病毒通过感染 Heliothis subflexa(BCIRL-HsAM1)细胞系并分离出几个克隆进行了噬菌斑纯化。根据 PstI、BstEII、StyI、HindIII 限制图谱估计,该病毒的平均估计基因组大小为 106 +/- 2.5 kbp(平均值+/-SE)。一个被命名为 TnMNPV/CmBCL9 的克隆被用于针对几种鳞翅目害虫的生物测定,并与 AcMNPV、AgMNPV、AfMNPV、PxMNPV 和 HzSNPV 进行了比较研究,这些病毒分别来自棉铃虫、银纹夜蛾、斜纹夜蛾、小菜蛾和玉米螟。感染性研究表明,TnMNPV/CmBCL9 对 Heliothis subflexa 和 T. ni 具有高度感染力,在这两种物种中的 LC(50)值均为 0.07 个封闭体/mm(2),对玉米螟和 Heliothis virescens 的感染力分别为 0.22 和 0.27 个封闭体/mm(2)。对分离株和选定的杆状病毒进行的限制性内切酶分析显示,其图谱与 AfMNPV 非常相似,但与其他杆状病毒的限制性内切酶图谱不同。杂交研究表明,TnMNPV/CmBCL9 与 AfMNPV 和 AcMNPV-HPP 密切相关。进一步的支持来自于使用分裂图网络对 TnMNPV/CmBCL9 的 polh、egt 和 p10 基因与其他杆状病毒的基因进行的系统发育分析,表明该病毒与 AcMNPV 变体、AfMNPV 和 Rachiplusia ou 的 RoMNPV 密切相关。