A RhoGAP venom protein from Microplitis mediator suppresses the cellular response of its host Helicoverpa armigera.

Female parasitoid wasps normally inject virulence factors together with eggs into their host to counter host immunity defenses. A newly identified RhoGAP protein in the venom of Microplitis mediator compromises the cellular immunity of its host, Helicoverpa armigera. RhoGAP1 proteins entered H. armigera hemocytes, and the host cellular cytoskeleton was disrupted. Depletion of MmGAP1 by injection of dsRNA or antibody increased the wasp egg encapsulation rate. An immunoprecipitation assay of overexpressed MmGAP1 protein in a Helicoverpa cell line showed that MmGAP1 interacts with many cellular cytoskeleton associated proteins as well as Rho GTPases. A yeast two-hybrid and a pull-down assay demonstrated that MmGAP1 interacts with H. armigera RhoA and Cdc42. These results show that the RhoGAP protein in M. mediator can destroy the H. armigera hemocyte cellular cytoskeleton, restrain host cellular immune defense, and increase the probability of successful parasitism.

Identification and temporal expression profiles of cuticular proteins in the endoparasitoid wasp, Microplitis mediator.

Recently, parasitoid wasp species Microplitis mediator has evoked increasing research attention due to its possible use in the control of Lepidoptera insects. Because insect development involves changes in cuticle composition, identification and expression analysis of M. mediator cuticular proteins may clarify the mechanisms involved in parasite development processes. We found 70 cuticular proteins from the M. mediator transcriptome and divided them into seven distinct families. Expression profiling indicated that most of these cuticular protein genes have expression peaks specific for one particular developmental stage of M. mediator. Eggs and pupae have the highest number of transcriptionally active cuticular protein genes (47 and 52 respectively). Only 12 of these genes maintained high expression activity during late larval development. Functional analysis of two larval proteins, MmCPR3 and MmCPR14, suggested their important role in the proper organization of the cuticle layers of larvae. During M. mediator larval development, normal cuticle formation can be supported by a limited number of cuticular proteins.

Insights into the venom protein components of Microplitis mediator, an endoparasitoid wasp.

Endoparasitoid wasps deliver a variety of maternal factors, such as venom proteins, viruses, and virus-like particles, from their venom and calyx fluid into hosts and thereby regulate the hosts' immune response, metabolism and development. The endoparasitoid, Microplitis mediator, is used as an important biological agent for controlling the devastating pest Helicoverpa armigera. In this study, using an integrated transcriptomic and proteomic analysis approach, we identified 75 putative venom proteins in M. mediator. The identified venom components were consistent with other known parasitoid wasps' venom proteins, including metalloproteases, serine protease inhibitors, and glycoside hydrolase family 18 enzymes. The metalloprotease and serpin family showed extensive gene duplications in venom apparatus. Isobaric tags for relative and absolute quantitation (iTRAQ) based quantitative proteomics revealed 521 proteins that were differentially expressed at 6 h and 24 h post-parasitism, including 10 wasp venom proteins that were released into the host hemolymph. Further analysis indicated that 511 differentially expressed proteins (DEP) from the host are primarily involved in the immune response, material metabolism, and extracellular matrix receptor interaction. Taken together, our results on parasitoid wasp venoms have the potential to enhance the application of endoparasitoid wasps for controlling insect pest.

A Metalloprotease Homolog Venom Protein From a Parasitoid Wasp Suppresses the Toll Pathway in Host Hemocytes.

Parasitoid wasps depend on a variety of maternal virulence factors to ensure successful parasitism. Encapsulation response carried out by host hemocytes is one of the major host immune responses toward limiting endoparasitoid wasp offspring production. We found that VRF1, a metalloprotease homolog venom protein identified from the endoparasitoid wasp, Microplitis mediator, could modulate egg encapsulation in its host, the cotton bollworm, Helicoverpa armigera. Here, we show that the VRF1 proenzyme is cleaved after parasitism, and that the C-terminal fragment containing the catalytic domain enters host hemocytes 6 h post-parasitism. Furthermore, using yeast two-hybrid and pull-down assays, VRF1 is shown to interact with the H. armigera NF-κB factor, Dorsal. We also show that overexpressed of VRF1 in an H. armigera cell line cleaved Dorsal in vivo. Taken together, our results have revealed a novel mechanism by which a component of endoparasitoid wasp venom interferes with the Toll signaling pathway in the host hemocytes.