Plant secondary metabolites as potential bioinsecticides? Study of the effects of plant-derived volatile organic compounds on the reproduction and behaviour of the pest beetle Tenebrio molitor.

Modern agriculture has many environmental consequences, such as soil contamination, accumulation of toxic compounds in the environment or risk of adverse effects on nontarget organisms and for these reasons, scientists are seeking a more environmentally friendly alternative to synthetic insecticides. This study investigated the effects of four plant secondary metabolites classified as volatile organic compounds (VOCs), which have potential as bioinsecticides, (E)-2-decenal, furfural, 2-undecanone and (E,E)-2-4-decadienal, in concentrations 10-5 and 10-7 M, on female reproductive processes and larval hatchability of the Tenebrio molitor beetle. Our study indicates proper development of ovaries after application of compounds however the volume of terminal oocytes was significantly reduced, with the strongest effect of (E)- 2-decenal which reduced the volume approximately three times. The relative vitellogenin expression level was reduced, with the strongest effect observed after application of furfural, (E,E)- 2-4-decadienal and (E)- 2-decenal in concentration 10-7 M, at the same time patency index was significantly reduced up to 2-times after application of furfural at 10-7 M. What is more important morphological changes translated into physiological ones. The number of laid eggs was affected, with the strongest inhibition after application of furfural (∼43% reduction), (E,E)- 2-4-decadienal (∼33%) and (E)- 2-decenal at concentration 10-7 M (∼33%). Moreover, we observed up to 13% (in case of 2-undecanone) decrease in larval hatchability. Tested compounds exhibited a repellent effect and caused 60% reduction of insect survivability after (E)- 2-decenal at concentration 10-5 M. Altogether, VOCs seems like potential bioactive compounds in plant protection.

Functional homology of tachykinin signalling: The influence of human substance P on the immune system of the mealworm beetle, Tenebrio molitor L.

Tachykinin-related peptides (TRPs) are one of the most prominent families of neuropeptides in the animal kingdom. Insect TRPs display strong structural and functional homology to vertebrate tachykinins (TKs). To study functional homologies between these two neuropeptide families, the influence of human substance P (SP, one of the essential vertebrate TKs) on the immune system of the mealworm beetle, Tenebrio molitor L., was analysed. Human SP influences the phagocytic abilities of T. molitor haemocytes. Peptide injection leads to an increase in the number of haemocytes participating in the phagocytosis of latex beads. In contrast, incubation of haemocytes from non-injected beetles in a solution of physiological saline and SP causes a decrease in phagocytic activity. Treatment with human SP also led to increased adhesion of haemocytes, but no changes in the arrangement of the F-actin cytoskeleton were observed. Interestingly, 6 h after human SP injection, increased DNA integrity in T. molitor haemocytes was reported. The opposite effects were observed 24 h after SP injection. Human SP caused the upregulation of humoral immune responses, such as phenoloxidase (PO) activity in the T. molitor haemolymph, and the downregulation of immune-related genes encoding coleoptericin A, tenecin 3 and Toll receptor. However, genes encoding attacin 2 and cecropin were upregulated. Despite these differences, the antimicrobial activity of T. molitor haemolymph was significantly lower in beetles injected with SP than in control beetles. Moreover, an analysis of the direct influence of SP on lysozyme activity was performed. Our results suggest that SP at a concentration of 10-6 M can directly inhibit lysozyme activity. However, an opposite effect was reported after the application of SP at a concentration of 10-4 M. The presented results suggest structural and functional homology between TK signalling in vertebrates and insects. Primarily, this was visible in the context of the humoral response and general antimicrobial activity of T. molitor haemolymph. However, some of the results related to haemocyte function may also indicate the importance of the TK and TRP sequences for evoking immunological effects.

A possible role of tachykinin-related peptide on an immune system activity of mealworm beetle, Tenebrio molitor L.

Tachykinin-related peptides (TRPs) are important neuropeptides. Here we show that they affect the insect immune system, especially the cellular response. We also identify and predict the sequence and structure of the tachykinin-related peptide receptor (TRPR) and confirm the presence of expression of gene encoding TRPR on Tenebrio molitor haemocytes. After application of the Tenmo-TRP-7 in T. molitor the number of circulating haemocytes increased and the number of haemocytes participating in phagocytosis of latex beads decreased in a dose- and time-dependent fashion. Also, Tenmo-TRP-7 affects the adhesion ability of haemocytes. Six hours after injection of Tenmo-TRP-7, a decrease of haemocyte surface area was observed under both tested Tenmo-TRP-7 concentrations (10-7 and 10-5 M). The opposite effect was reported 24 h after injection, which indicates that the influence of Tenmo-TRP-7 on modulation of haemocyte behaviour differs at different stages of stress response. Tenmo-TRP-7 application also resulted in increased phenoloxidase activity 6 and 24 h after injection. The assessment of DNA integrity of haemocytes showed that the injection of Tenmo-TRP-7 at 10-7 M led to a decrease in DNA damage compared to control individuals. This effect was only visible 6 h after Tenmo-TRP-7 application. After 24 h, Tenmo-TRP-7 injection increased DNA damage. We also confirmed the expression of immune-related genes in nervous tissue of T. molitor. Transcripts for genes encoding receptors participating in pathogen recognition processes and antimicrobial peptides were detected in T. molitor brain, retrocerebral complex and ventral nerve cord. These results may indicate a role of the insect nervous system in pathogen recognition and modulation of immune response similar to vertebrates. Taken together, our results support the notion that tachykinin-related peptides probably play an important role in the regulation of the insect immune system. Moreover, some resemblances with action of tachykinin-related peptides and substance P showed that insects can be potential model organisms for analysis of hormonal regulation of conserved innate immune mechanisms.

New activity of yamamarin, an insect pentapeptide, on immune system of mealworm, Tenebrio molitor.

In insects, two types of the immune responses, cellular and humoral, constitute a defensive barrier against various parasites and pathogens. In response to pathogens, insects produce a wide range of immune agents that act on pathogens directly, such as cecropins or lysozyme, or indirectly by the stimulation of hemocyte migration or by increasing phenoloxidase (PO) activity. Recently, many new immunologically active substances from insects, such as peptides and polypeptides, have been identified. Nevertheless, in the most cases, their physiological functions are not fully known. One such substance is yamamarin - a pentapeptide isolated from the silk moth Antheraea yamamai. This yamamarin possesses strong antiproliferative properties and is probably involved in diapause regulation. Here, we examined the immunotropic activity of yamamarin by testing its impact on selected functions of the immune system in heterologous bioassays with the beetle Tenebrio molitor, commonly known as a stored grains pest. Our results indicate that the pentapeptide affects the activity of immune processes in the beetle. We show that yamamarin induces changes in both humoral and cellular responses. The yamamarin increases the activity of PO, as well as causes changes in the hemocyte cytoskeleton and stimulates phagocytic activity. We detected an increased number of apoptotic hemocytes, however after the yamamarin injection, no significant variations in the antibacterial activity in the hemolymph were observed. The obtained data suggest that yamamarin could be an important controller of the immune system in T. molitor.