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One of the interesting features of  Anticarsia gemmatalis multiple nucleopolyhedrovirus  isolate 2D (AgMNPV-2D) genome is the absence of chitinase ( chiA ) and cathepsin ( v - cath ) genes. This characteristic may be responsible for the lack of liquefaction and melanization in  A. gemmatalis  larvae killed by AgMNPV-2D infection. This study aimed to test the hypothesis that CHIA and V-CATH proteins from  Choristonera fumiferana  DEF  multiple nucleopolyhedrovirus  (CfDEFNPV) are able to liquefy and melanize the cuticle of  A. gemmatalis  larvae infected by a recombinant AgMNPV containing  chiA  and  v - cath  genes inserted in its genome. A fragment from the CfDefNPV genome containing  chiA  and  v - cath  genes was inserted into the genome of AgMNPV-2D. The recombinant virus (vAgp2100Cf.chiA/v-cath) was purified and used to infect insect cells and larvae. Transcripts of  v - cath  and  chiA  genes were detected along the infection of insect cells by qRT-PCR, from early to late phases of infection. The analysis of  A. gemmatalis  larvae killed by vAgp2100Cf.chiA/v-cath infection confirmed the hypothesis proposed. The vAgp2100Cf.chiA/v-cath showed higher insecticidal activity against third instar  A. gemmatalis  larvae when compared to AgMNPV-2D. The mean time to death was also lower for the vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D at 10 days post infection. Occlusion body production was higher in  A. gemmatalis  larvae infected with vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D. Enzyme assays showed higher chitinase and cysteine protease activities in insect cells and insects infected with vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D. The introduction of  chiA  and  v-cath  genes into the genome of AgMNPV improves its insecticidal activity against  A. gemmatalis  larvae and this recombinant virus could be used as an alternative to the wild type virus to control this important insect pest.

plos one

A Recombinant Anticarsia gemmatalis MNPV Harboring chiA and v-cath Genes from Choristoneura fumiferana Defective NPV Induce Host Liquefaction and Increased Insecticidal Activity