Prophenoloxidase (PPO) plays an important role in melanization, necessary for defense against intruding parasitoids. Parasitoids have evolved to inject maternal virulence factors into the host hemocoel to suppress hem...Prophenoloxidase (PPO) plays an important role in melanization, necessary for defense against intruding parasitoids. Parasitoids have evolved to inject maternal virulence factors into the host hemocoel to suppress hemolymph melanization for the successful development of their progeny. In this study, the full-length complementary DNA (cDNA) of a Pieris rapae PPO was cloned. Its cDNA contained a 2 076-base pair (bp) open reading frame (ORF) encoding 691 amino acids (aa). Two putative copper-binding sites, a proteolytic activation site, three conserved hemocyanin domains, and a thiol ester motif were found in the deduced amino acid sequence. According to both multiple alignment and phylogenetic analysis, P. rapae PPO gene cloned here is a member of the lepidopteran PPO-2 family. Injection of Cotesia glomerata venom or calyx fluid resulted in reduction of P. rapae hemolymph phenoloxidase activity, demonstrating the ability to inhibit the host's melanization. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) showed that transcripts of P. rapae PPO-2 in the haemocytes from larvae had not significantly changed following venom injection, suggesting that the regulation of PPO messenger RNA (mRNA) expression by venom was not employed by C. glomerata to cause failure of melanization in parasitized host. While decreased P. rapae PPO-2 gene expression was observed in the haemocytes after calyx fluid injection, no detectable transcriptional change was induced by parasitization, indicating that transcriptional down-regulation of PPO by calyx fluid might play a minor role involved in inhibiting the host's melanization.展开更多
Endoparasitoid wasps can develop inside permissive host due to their ability to overcome or to evade the host’s cellular and humoral immune response. Oviposition of Campoletis chlorideae (Hymenoptera: Ichneumonidae) ...Endoparasitoid wasps can develop inside permissive host due to their ability to overcome or to evade the host’s cellular and humoral immune response. Oviposition of Campoletis chlorideae (Hymenoptera: Ichneumonidae) in larvae of Helicoverpa armigera (Lepidoptera) was accompanied by inhibition of phenoloxidase (PO) activity and melanization reaction in host hemolymph in vitro. PO activity in host plasma was decreased about 83% 48 h post para-sitization. A similar result was found when the host insect was injected with 0.5 wasp equivalent calyx fluid. This indicated that the calyx fluid was concerned with suppression of PO activity after parasitization. Furthermore, the prophen-oloxidase (proPO) in host haemocytes could be activated by bovine trypsin in unparasitized insects, while it could not be activated in parasitized or calyx fluid-injected host. The results suggested that inhibition of PO activity by parasitization was related to the calyx fluid of Campoletis chlorideae, and the components of calyx展开更多
基金supported by the Applied Basic Research Program of Yunnan Province (No. 2010CD063)the Science Foundation of Southwest Forestry University (No. 110903)+2 种基金the Science Foundation of the Department of Education of Yunnan Province (No. 2010Y294)the Open Foundation of Key Laboratory of Forest Disaster Warning and Control of Yunnan Province (No. ZK09A101)the Key Subject of Forest Protection of Yunnan Province (No. XKZ200905),China
文摘Prophenoloxidase (PPO) plays an important role in melanization, necessary for defense against intruding parasitoids. Parasitoids have evolved to inject maternal virulence factors into the host hemocoel to suppress hemolymph melanization for the successful development of their progeny. In this study, the full-length complementary DNA (cDNA) of a Pieris rapae PPO was cloned. Its cDNA contained a 2 076-base pair (bp) open reading frame (ORF) encoding 691 amino acids (aa). Two putative copper-binding sites, a proteolytic activation site, three conserved hemocyanin domains, and a thiol ester motif were found in the deduced amino acid sequence. According to both multiple alignment and phylogenetic analysis, P. rapae PPO gene cloned here is a member of the lepidopteran PPO-2 family. Injection of Cotesia glomerata venom or calyx fluid resulted in reduction of P. rapae hemolymph phenoloxidase activity, demonstrating the ability to inhibit the host's melanization. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) showed that transcripts of P. rapae PPO-2 in the haemocytes from larvae had not significantly changed following venom injection, suggesting that the regulation of PPO messenger RNA (mRNA) expression by venom was not employed by C. glomerata to cause failure of melanization in parasitized host. While decreased P. rapae PPO-2 gene expression was observed in the haemocytes after calyx fluid injection, no detectable transcriptional change was induced by parasitization, indicating that transcriptional down-regulation of PPO by calyx fluid might play a minor role involved in inhibiting the host's melanization.
基金This work was supported by a special fund for the Major State Basic Research Project of China (Grant No. 2000016208) and the National Natural Science Foundation of China (Grant No. 30070517).
文摘Endoparasitoid wasps can develop inside permissive host due to their ability to overcome or to evade the host’s cellular and humoral immune response. Oviposition of Campoletis chlorideae (Hymenoptera: Ichneumonidae) in larvae of Helicoverpa armigera (Lepidoptera) was accompanied by inhibition of phenoloxidase (PO) activity and melanization reaction in host hemolymph in vitro. PO activity in host plasma was decreased about 83% 48 h post para-sitization. A similar result was found when the host insect was injected with 0.5 wasp equivalent calyx fluid. This indicated that the calyx fluid was concerned with suppression of PO activity after parasitization. Furthermore, the prophen-oloxidase (proPO) in host haemocytes could be activated by bovine trypsin in unparasitized insects, while it could not be activated in parasitized or calyx fluid-injected host. The results suggested that inhibition of PO activity by parasitization was related to the calyx fluid of Campoletis chlorideae, and the components of calyx
