Parasitization by Macrocentrus cingulum (Hymenoptera: Braconidae) influences expression of prophenoloxidase in Asian corn borer Ostrinia furnacalis.

A prophenoloxidase (PPO) cDNA (OfPPO) was cloned from the Asian corn borer Ostrinia furnacalis. Sequence analysis revealed a full length transcript of the OfPPO cDNA with 2,686 bp, containing a 2,079 bp open reading frame (ORF), a 73-bp 5'-untranslated region, and a 534-bp 3'-untranslated region with a poly(A) signal. The ORF encodes a 693-amino acid polypeptide, containing two distinct copper-binding regions, a plausible thiol ester site, two proteolytic activation sites, and a conserved C-terminal region, but lacks a signal peptide sequence. Expression of the OfPPO transcript in the plasma, hemocytes, fat body and midgut was inhibited by Macrocentrus cingulum at 4 h post-parasitization (pp). In situ hybridization analysis showed that the hemocytes, especially the oenocytoids, hybridized strongly with the DNA probes of the OfPPO gene. No signal was detected in the cuticular epithelium or fat body of the parasitized larvae. Colloidal gold particles were used to visualize the PPO by immunoelectron microscopy. The time course study revealed a decrease in the labeling of the OfPPO at 4, 6, 8, 12, and 1 day pp in the larval integument and midgut parasitized by M. cingulum. We infer from time course studies of OfPPO gene expression and PO enzymatic activity that OfPPO in the integument is released from hemocytes and that the OfPPO expression was influenced at the transcriptional, translational, and then the post-translational level by parasitization challenge.

Wheat germ oil in larval diet influences gene expression in adult oriental fruit fly.

Culture medium supplemented with wheat germ oil (WGO) causes physiological reactions, such as increased fecundity and mobility, in some insects. Although the impact of WGO on insect physiology is important, the mechanisms of these actions are poorly understood. In this paper, we test the hypothesis that the addition of WGO to medium developed for larval oriental fruit flies modulates gene expression in the corresponding adults. We separately reared larvae of Bactrocera dorsalis on diets lacking or supplemented with WGO, and analyzed for expressed proteins in the resulting adult males and females by 2D-electrophoresis. Analysis of the gels revealed significant changes in expression levels of >70 proteins, 64 of which were identified by mass spectrometric analysis on MALDI-TOF/TOF. Apparent changes in expression levels for 6 of these proteins were confirmed by quantitative real-time PCR, showing that the changes in mRNA expression were reflected in changes in protein expression. These findings support the hypothesis that one mechanism of WGO actions in insect nutrition is the modulation of gene expression.