Proteinase activity in the midgut of the pentatomid stinkbug Podisus maculiventris was investigated. The optimal pH for adult and nymph proteolysis was pH 6.0 and pH 6.5, respectively. Proteinase activity was characterised using a range of diagnostic inhibitors. Activity of both adult and nymphal gut extracts, detected by the hydrolysis of Z-Phe-Arg-pNA, was inhibited to <20% of control levels by several inhibitors (e.g. E-64 and chicken egg white cystatin) associated with the inhibition of cysteine proteinases. The less specific inhibitor leupeptin reduced proteolytic activity to around 1.0% of the control values. In-gel analysis of the enzymes revealed that proteolytic activity was due to at least four proteinases, of ca. 30, 36, 50 and 110 kDa, which were all susceptible to E-64 inhibition. Salivary gland extracts gave maximal activity at pH 8.0 when tested for general proteolytic activity using fluorescent BODIPY-FL casein substrate, and showed moderate levels of inhibition when incubated with inhibitors of serine-, cysteine-, aspartic- and metallo-proteinases. Leupeptin and PMSF gave the highest levels of inhibition of salivary proteolytic activity, at ca. 50%, whilst the plant-derived inhibitors SKTI, CpTI and OC-1 did not inhibit proteolysis.
The nature and rate of juvenile hormone (JH) biosynthesis and effects of allatostatins and allatotropin have been investigated in isolated corpora allata (CA) of adults and larvae of the noctuid tomato moth, Lacanobia oleracea. In adult female CA, mean rates of synthesis were relatively constant (10-16 pmol/pr/h) at all times. However, the range of JH synthesis by individual CA of similarly aged insects was quite large (2-30 pmol/pr/h). High performance liquid chromatography (HPLC) separation shows that adult female moth CA synthesise predominantly JH I and JH II. Rates of JH synthesis in vitro are dependent on methionine concentration. Synthetic Manduca sexta allatostatin (Mas-AS) caused a dose-dependent inhibition of JH synthesis by adult female CA but only to a max. of 54%, whilst 10 µM synthetic M. sexta allatotropin caused a 37% stimulation of CA activity. At 1 mM the cockroach allatostatin, Dip-allatostatin-2, had no significant effect on JH synthesis. In larval L. oleracea, rates of JH biosynthesis were very low., Neil Audsley, Robert J. Weaver, John P. Edwards, and Lit
The comparative biology of the solitary endoparasitoid Meteorus gyrator (Thun.) was investigated in five noctuid pest species. Meteorus gyrator parasitized all larval stages of the noctuid pests Lacanobia oleracea, Mamestra brassicae, Spodoptera exigua, Spodoptera littoralis and Chrysodeixis chalcites. When female parasitoids were offered host larvae of all developmental stages, host larvae in their third stadium were parasitized most frequently in all species. When the parasitoid was offered a choice of third stadium larvae from each of the five lepidopteran host species, L. oleracea was the most frequently parasitized, followed by M. brassicae. The growth of L. oleracea and M. brassicae was markedly reduced when larvae were parasitized by M. gyrator, with the effect of parasitism on the remaining species being much less pronounced. In excess of 94% of parasitized hosts gave rise to live wasps in L. oleracea, whilst in M. brassicae only 52% of parasitized hosts gave rise to a live wasp, the remainder dying (44.6%) or pupating (3.1%).
The larval development of the parasitoid was also strongly influenced by the host species. Thus, the development (egg to cocoon) of female wasps was most rapid in C. chalcites (9.7 ± 0.09 days), and longest in M. brassicae (17.2 ± 1.08 days). The cocoon weight of wasps was also significantly affected by host species, with the heaviest female cocoons being derived from C. chalcites (10.4 ± 0.16 mg) and the lightest female cocoons being derived from M. brassicae (7.0 ± 0.14 mg). In all cases, the development times and cocoon weights of male parasitoids were less than those of female wasps.