We identified a new member of the Tetraviridae, Providence virus (PrV), persistently infecting a midgut cell line derived from the corn earworm (Helicoverpa zea). Virus purified from these cells also productively infected a H. zea fat body cell line, and a cell line from whole embryos of the beet armyworm, Spodoptera exigua. PrV is thus the first tetravirus shown to replicate in cell culture. PrV virions are isometric particles composed of two structural proteins (60 and 7.4 kDa) that encapsidate both the genomic (6.4 kb) and the subgenomic (2.5 kb) RNAs. The monopartite organization of the PrV genome resembles that of Nudaurelia beta virus and Thosea asigna virus, members of the genus Betatetravirus. The predicted sequence of the PrV structural proteins demonstrates homology to tetraviruses in both genera. The infectivity of PrV for cultured cells uniquely permitted examination of tetravirus RNA and protein synthesis during synchronous infection. The discovery of PrV greatly facilitates studies of tetravirus molecular biology.
This study describes a new baculovirus isolate recovered from infected larvae of the diamondback moth, Plutella xylostella (L.), and identified as a multiple nucleopolyhedrovirus (MNPV). The plaque purified isolate designated as PxMNPVCL3 was found to be pathogenic to P. xylostella, Heliothis virescens (F.), Trichoplusia ni (Hübner), H. subflexa (Guenée), Helicoverpa zea (Boddie), Spodoptera exigua (Hübner), and S. frugiperda (J. E. Smith) larvae in decreasing order of susceptibility. The LC50 for diamondback moth, the most susceptible, was 6 occlusion bodies (OB)/cm2, whereas the most resistant species, namely S. frugiperda, was 577 OB/cm2. PxMNPVCL3 was more pathogenic to diamondback moth by 3-4 log cycles as compared with 2 broad-spectrum baculoviruses, namely Autographa california (alfalfa looper) MNPV and Anagrapha falcifera (celery looper) MNPV. The 3 baculoviruses were compared with each other and characterized by restriction endonuclease (REN) analysis, hybridization, and neutralization tests. Fragmentation profiles generated by REN showed that the 3 baculoviruses shared some fragments in common. Hybridization studies employing digoxigenin labeled PxMNPVCL3 DNA as a probe revealed the close but distinct relationship of these 3 viruses. Neutralization tests confirmed the hybridization studies, namely that the 3 viruses although genetically similar are distinguishable from each other.
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