Based on these induction and knockdown analyses, they concluded that IMD-dependent AMP genes were induced by gram negative bacteria while one Toll-dependent AMP gene was induced by gram positive bacteria (Bs that possesses DAP-type PG and gram-negative Flavobacterium sp.). These inconsistencies between their and our results may be attributed somewhat to differences of experimental conditions mentioned above. Zou et al. used Ml, Ec, Sc and a pathogenic fungus Candida albicans as elicitors, measured induction of T. castaneum AMP mRNAs by qRT-PCR and presented a schematic illustration of gene induction in their paper [39]. In this illustration,

they showed that Att2, Cec3, Col1, Def1 and Def2 mRNAs were dramatically induced in adults 24 h after microbial challenge and Ml-challenged animals showed BMS-777607 cell line this website the strongest induction of these mRNAs. Our data showed of the three microbes, Ec was the most potent elicitor for seven AMP genes out of the nine tested

genes when examined with pupae at 24 h post microbial challenge, whereas Ml acted as a potent elicitor comparable to Ec at 6 h. The remaining two (Att3 and Def1 constituting group IV) were not induced well by any of the five microbes at the two time points tested. The inconsistencies may also be ascribed to differences in experimental methods such as developmental stages of animals and microbe handing. They used adult beetles and pricked them with needles dipped in PBS containing the microbes. Our experimental procedures may provide more

accurate profiles of AMP induction Liothyronine Sodium because we challenged animals with microbes prepared at defined concentrations. Our study showed that the basal mRNA amounts of group IV genes Att3 and Def1 were relatively low compared to other AMP mRNA, and their induction by the five microbes was very weak or negligible. These low basal levels may be accounted for by tissue- or stage-specific induction of these AMP genes. In Drosophila, tissue- and stage-specific AMP gene induction has been reported using transgenic fly lines expressing green fluorescent protein under the control of AMP gene promoters [43]. According to their results, induction profiles of some Drosophila AMP genes, such as Cecropin, Defensin and Attacin, were highly tissue-, developmental stage- and sex-specific. In this study, total RNA was extracted from the whole body pupae and used as a template for qRT-PCR. When we assume that Tribolium Att3 and Def1 are induced only in a small portion of pupal tissues and basal low level expression occurs in a wider variety of tissues, the observed induction levels may be very low or negligible. Zou et al. annotated T. castaneum components related to immune reaction [39]. Their study showed that T.