The diapause inducement condition, cold hardiness, and flight ability in Cry1Ac-resistant (BtR) and Cry1Ac-susceptible (96S) strains of Helicoverpa armigera (Hübner) were compared in the laboratory. The BtR strain was derived from the 96S strain and shows 1375-fold resistance to the Cry1Ac toxin after having been selected for 52 generations. Compared with the 96S strain, the Bt-resistant strain was more likely to go into diapause under a short-photoperiod environment. At 11L : 13D, 12L : 12D and 13L : 11D photoperiods, the percentages of BtR insects entering diapause were 72.7%, 82.9% and 68.7%, respectively, which were significantly higher than those in the 96S strain (58.6%, 67.4% and 46.3%, respectively) under the same conditions. The supercooling points (SCP) and freezing points (FP) were not significantly different between the BtR and 96S strains. The LT50 (50% lethal time) and LT90 (90% lethal time) of BtR pupae were also not significantly different from those of the 96S strain at -15°C. The moths from both strains had similar flight ability when their larvae were fed with nontoxic control diet. However, the total flight distance of these BtR moths was 56.2 km whose larvae fed on normal diet, which was more than twice as much as for those feeding on Bt diet (26.2 km). Flight duration for these BtR moths was longer after feeding on normal diet (11.6 h) than after feeding on Bt diet (7.3 h).
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.