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A recombinant  Helicoverpa armigera  nucleopolyhedrovirus (HearNPV), Ar1b-HearNPV, was constructed and identified as an improved bio-control agent of  Helicoverpa armigera  larvae. The HearNPV  polyhedrin  promoter was used to express the insect-specific neurotoxin gene,  ar1b , which was originally isolated from the Australian funnel-web spider ( Atrax robustus ). RT-PCR and Western blotting analysis showed that both the  ar1b  transcript and protein were produced successfully in Ar1b-HearNPV-infected HzAM1 cells. In order to investigate the influence of foreign gene insertion in HearNPV, including the  ar1b  gene, chloramphenicol resistance gene,  lacZ , kanamycin resistance gene, and the gentamicin resistance gene, two virus strains (HZ8-HearNPV and wt-HearNPV) were used as controls in the cell transfection analysis. As expected, foreign gene insertion had no impact on budded virus production and viral DNA replication. Both optical microscopy and electron microscopy observations indicated that the formation of the occlusion bodies of recombinant virus was similar to wild type virus. The Ar1b-HearNPV-infected  H .  armigera  larvae exhibited paralysis and weight loss before dying. This recombinant virus also showed a 32.87% decrease in LT 50  assays compared with the wild type virus. Besides, Ar1b-HearNPV also inhibited host larval growth and diet consumption. This inhibition was still significant in the older instar larvae treated with the recombinant virus. All of these positive properties of this novel recombinant HearNPV provide a further opportunity to develop this virus strain into a commercial product to control the cotton bollworm.

plos one

A Novel Neurotoxin Gene ar1b Recombination Enhances the Efficiency of Helicoverpa armigera Nucleopolyhedrovirus as a Pesticide by Inhibiting the Host Larvae Ability to Feed and Grow