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.
In vitro embryogenic callus is a critical factor for genetic transformation of rice, especially for indica varieties. In this study, we investigated the relationship between polyamines, including putrescine (Put), spermidine (Spd) and spermine (Spm), and callus browning, and we studied the effect of exogenous Put on callus regeneration and on the content of endogenous polyamines. In addition, the expression levels of arginine decarboxylase gene (Adcl) and S-adenosylmethionine decarboxylase gene (Samdc) in embryogenic callus were studied by quantitative Real-time PCR analysis. The results showed that the contents of endogenous Put and Spd in the browning callus were significantly lower than those in normal callus. Exogenous Put could effectively improve the growing state of callus of indica rice and enhance the development of embryogenic callus. The content of endogenous polyamines in embryogenic callus, especially Spd and Spm, was increased after addition of exogenous Put. Additionally, exogenous Put also had an obvious impact on the expression levels of Adcl but partial effect on the expression levels of Samdc gene. This study could increase the knowledge of both embryogenic callus induction and polyamine catabolism in callus in indica rice.
Arbuscular mycorrhizal fungi are the most widespread root fungal symbionts, forming associations with the vast majority of plant species. Ectomycorrhizal development alters gene expression in plant symbionts. In this work we examined the effect of arbuscular mycorrhizal fungi spores on the growth and development of Brassica and on the expression of BnMT2 in winter rape. In a pot experiment, rape seedlings growing on different types of sterile and nonsterile soils were inoculated simultaneously with mycorrhizal fungi spores of Acaulospora longula,Glomus geosporum, Glomus mosseae and Scutellospora calospora. As compared with control plants growing in the absence of spores, ten-week-old seedlings of Brassica napus L. in sterile soil inoculated with arbuscular spores had longer shoots and higher fresh biomass of above-ground plant parts. In other types of substrates enriched with mycorrhizal fungi spores, the plants were smaller than non-inoculated plants. The presence of AMF spores stimulated the elongation growth of hypocotyls in both analyzed substrates. BnMT2 expression was highest in plants growing on the sterile substrate. Generally, the presence of mycorrhizal fungi spores appeared to have an adverse effect on the growth of rape plants.
This paper presents methods for optimal test frequencies search with the use of heuristic approaches. It includes a short summary of the analogue circuits fault diagnosis and brief introductions to the soft computing techniques like evolutionary computation and the fuzzy set theory. The reduction of both, test time and signal complexity are the main goals of developed methods. At the before test stage, a heuristic engine is applied for the principal frequency search. The methods produce a frequency set which can be used in the SBT diagnosis procedure. At the after test stage, only a few frequencies can be assembled instead of full amplitude response characteristic. There are ambiguity sets provided to avoid a fault tolerance masking effect.