Protein digestion in insects relies on several groups of proteases, among which trypsin plays a prominent role. In the current study, larvae of Pieris brassicae L. were fed radish leaves treated with 1 mM concentrations of three specific inhibitors of trypsin: AEBSF.HCl [4-(2- -aminoethyl)-benzenesulfonyl fluoride, monohydrochloride], TLCK (N-a-tosyl-l-lysine chloromethyl ketone) and SBTI (Soybean Trypsin Inhibitor) to find their potential effects on gene expression of trypsin. Initially, RT-PCR analysis revealed a gene of 748 bp responsible for synthesizing the digestive trypsin in P. brassicae larvae. Also, qRT-PCR data indicated a statistically greater expression of trypsin gene in the larvae fed 1 mM concentrations of AEBSF.HCl, TLCK and SBTI than the control. Results of the current study indicated that synthetic inhibitors can not only negatively affect the gene expression of P. brassicae trypsin, but also the insect can activate a compensatory mechanism against interruption of protein digestion by inducing more expression of the gene and producing more trypsin into the midgut lumen.
The effect of monoterpenoid 1,8-cineol on the toxicity and physiology of elm leaf beetle, Xanthogaleruca luteola Müller under laboratory conditions (26 ± 1°C, 65 ± 10% RH and 16L : 8D h) was investigated. Initially, LC30 and LC50 values of the constituent were estimated to be 23.5 ppm and 31.9 ppm for the last instar larvae after 48 h, respectively. Significant changes were observed in the values of relative growth rate (RGR), efficiency of conversion of ingested food (ECI), efficiency of conversion of digested food (ECD), approximate digestibility (AD) and consumption index (CI) between control and treated larvae with 1,8-cineol. The amounts of protein, glucose and urea decreased in the treated larvae in comparison with control. Similar findings were observed in the activities of alkaline phosphatase and lactate dehydrogenase while the activities of glutathione S-transferase and esterase significantly increased in the treated larvae using CDNB and α-naphtyl acetates as the substrates. Morphological and histological changes brought about by 1,8-cineol in the present study are indicative of growth inhibition targeting specific organs such as those of reproduction. We believe that 1,8-cineol can be considered as a safe and environmentally friendly compound.