RNA m6 A Methylation Suppresses Insect Juvenile Hormone Degradation to Minimize Fitness Costs in Response to A Pathogenic Attack.

Bioinsecticides and transgenic crops based on the bacterial pathogen Bacillus thuringiensis (Bt) can effectively control diverse agricultural insect pests, nevertheless, the evolution of resistance without obvious fitness costs has seriously eroded the sustainable use of these Bt products. Recently, it has been discovered that an increased titer of juvenile hormone (JH) favors an insect host (Plutella xylostella) to enhance fitness whilst resisting the Bt pathogen, however, the underlying regulatory mechanisms of the increased JH titer are obscure. Here, the involvement of N6 -methyladenosine (m6 A) RNA modification in modulating the availability of JH in this process is defined. Specifically, it is found that two m6 A methyltransferase subunit genes, PxMettl3 and PxMettl14, repress the expression of a key JH-degrading enzyme JH esterase (JHE) to induce an increased JH titer, mitigating the fitness costs associated with a robust defense against the Bt pathogen. This study identifies an as-yet uncharacterized m6 A-mediated epigenetic regulator of insect hormones for maintaining fitness during pathogen defense and unveils an emerging Bt resistance-related m6 A methylation atlas in insects, which further expands the functional landscape of m6 A modification and showcases the pivotal role of epigenetic regulation in host-pathogen interactions.

Txp40, an insecticidal toxin protein from Xenorhabdus nematophila: Purification, toxicity assessment and biophysical characterization.

Txp40 is a ubiquitous toxin from Xenorhabdus and Photorhabdus bacteria, exhibits insecticidal activity against a wide range of insect pests belonging to Lepidoptera and Diptera orders. Initially, Txp40 affects midgut of the target insect and further damages some other tissues like fat bodies but the detailed mode of action is not known. Txp40 shares no significant sequence match to any proteins with known structure or function, suggesting that it is a novel type of insecticidal toxin. Here, we report purification, toxicity and biophysical characterization of the Txp40b toxin from X. nematophila (ATCC, 19061). The recombinant Txp40b was found toxic to Galleria mellonella larvae with LD50 of 30.42 ng larva-1. Circular dichroism spectroscopy revealed that purified Txp40b is an α-helix rich protein with a relatively lower melting temperature of 45 °C. In-silico model generated suggests two domain structure of Txp40b toxin. Detailed structural analysis of Txp40b will provide new insights about the mode of action and possibly it would illustrate a new domain and/or motif in the area of insecticidal proteins.

Pharmacological Properties of the Type 1 Tyramine Receptor in the Diamondback Moth, Plutella xylostella.

Tyramine receptors (TARs) can be activated by tyramine (TA) or octopamine (OA) and have been shown to be related to physiological regulation (e.g., gustatory responsiveness, social organization, and learning behavior) in a range of insect species. A tyramine receptor gene in Plutella xylostella, Pxtar1, was cloned and stably expressed in the HEK-293 cell line. Pharmacological properties and expression profile of Pxtar1 were also analyzed. Tyramine could activate the PxTAR1 receptor, increasing the intracellular Ca2+ concentration ((Ca2+)i) at an EC50 of 13.1 nM and reducing forskolin (10 µM)-stimulated intracellular cAMP concentration ((cAMP)i) at an IC50 of 446 nM. DPMF (a metabolite of amitraz) and L(-)-carvone (an essential oil) were found to act as PxTAR1 receptor agonists. Conversely, yohimbine and mianserin had significant antagonistic effects on PxTAR1. In both larvae and adults, Pxtar1 had the highest expression in the head capsule and expression of Pxtar1 was higher in male than in female reproductive organs. This study reveals the temporal and spatial differences and pharmacological properties of Pxtar1 in P. xylostella and provides a strategy for screening insecticidal compounds that target PxTAR1.

Development of reduced cost diets for the mass rearing of false codling moth (Thaumatotibia leucotreta).

BACKGROUND: Four trials were conducted; the first established optimal cooking time for false codling moth diets using a microwave oven while the other three trials aimed at determining nutrient requirements, testing these, and evaluating alternative cheaper raw materials. RESULTS: Microwave preparation for 8 min, using a 1000 W microwave oven was found to be the optimal cooking time for all treatment diets at 200 g kg-1 moisture. The minimum specification (MS) diet yielded more larvae per jar (mean ± standard deviation) (759 ± 29) than the control (CON) (652 ± 32) diet and the ideal amino acid profile (IAAP) diet (596 ± 46). The third trial subsequently utilized diets formulated according to the MS diet nutrient specifications but using novel and cheaper raw materials (D1 and D2). Production characteristics of diets D1 and D2 was compared to that of CON. Diets D1 and D2 yielded more larvae per jar (D1 = 911 ± 40; D2 = 830 ± 40) than that of CON (428 ± 40) with the added advantage that the newly formulated diets were cheaper. CONCLUSION: Reduced cost feeds were formulated, mixed and tested which resulted in substantial saving in the mass rearing of the species for integrated pest management. This method shows potential for diet development of other insect species as well. © 2019 Society of Chemical Industry.

Costs and benefits of plant allelochemicals in herbivore diet in a multi enemy world.

Sequestration of plant defensive chemicals by herbivorous insects is a way of defending themselves against their natural enemies. Such herbivores have repeatedly evolved bright colours to advertise their unpalatability to predators, i.e. they are aposematic. This often comes with a cost. In this study, we examined the costs and benefits of sequestration of iridoid glycosides (IGs) by the generalist aposematic herbivore, the wood tiger moth, Parasemia plantaginis. We also asked whether the defence against one enemy (a predator) is also effective against another (a parasitoid). We found that the larvae excrete most of the IGs and only small amounts are found in the larvae. Nevertheless, the amounts present in the larvae are sufficient to deter ant predators and also play a role in defence against parasitoids. However, excreting and handling these defensive plant compounds is costly, leading to longer development time and lower pupal mass. Interestingly, the warning signal efficiency and the amount of IGs in the larvae of P. plantaginis are negatively correlated; larvae with less efficient warning signals contain higher levels of chemical defence compounds. Our results may imply that there is a trade-off between production and maintenance of coloration and chemical defence. Although feeding on a diet containing IGs can have life-history costs, it offers multiple benefits in the defence against predators and parasitoids.

Sex pheromones in mate assessment: analysis of nutrient cost of sex pheromone production by females of the moth Heliothis virescens.

It has been postulated that sex pheromones, in addition to their role in mate recognition and/or finding, may also serve a role in assessment of mate quality. For this, a sex pheromone must give honest information about a signaler's quality, with honesty ensured by a direct metabolic or indirect fitness cost to the signaler. Using a stable isotope tracer-tracee method, we characterized the nutrient pools that fuel sex pheromone production in females of the moth Heliothis virescens, as well as the relative importance of larval- and adult-acquired nutrients to this process. Females used three pools for de novo biosynthesis of sex pheromone, hemolymph trehalose, glycogen (via trehalose) and fat, and produced ca. 25% of pheromone directly from stored (previously synthesized) precursor fatty acids. Pheromone was produced roughly equally from carbohydrate and fat. Adult feeding was very important for pheromone biosynthesis, with a maximum of 65% of de novo biosynthesized pheromone produced from a single adult feed (carbohydrate). Although these nutrient pools are shared with other reproductive physiologies, notably oocyte production, it is unlikely that pheromone production imposes a significant metabolic cost on females, because (i) the amount of nutrients used for pheromone production is negligible compared with that available, (ii) the hemolymph trehalose pool is readily replaceable throughout the adult life, and (iii) in mated females, carbohydrate shortages result in reduced allocation to pheromone.

Assessment of in vivo antimicrobial activity of the carbene silver(I) acetate derivative SBC3 using Galleria mellonella larvae.

The antimicrobial drug candidate 1,3-dibenzyl-4,5-diphenyl-imidazol-2-ylidene silver(I) acetate (SBC3) was evaluated for its ability to function in vivo using larvae of Galleria mellonella. A SBC3 concentration of 25 µg/ml inhibited the growth of Staphylococcus aureus by 71.2% and Candida albicans by 86.2% in vitro. Larvae inoculated with 20 µl of SBC3 solution showed no ill effects up to a concentration of 250 µg/ml but administration of 500 µg/ml resulted in a 40% reduction in larval survival and administration of a dose of 1,000 µg/ml resulted in total larval death at 24 h. Larvae inoculated with S. aureus or C. albicans and subsequently administered SBC3 showed increased survival. Administration of SBC3 to larvae did not boost the insect immune response as indicated by lack of an increase in the density of circulating haemocytes (immune cells). The abundance of a number of proteins involved in the insect immune response was reduced in larvae that received 20 µl SBC3 solution of 100 µg/ml. This is the first demonstration of the in vivo activity of SBC3 against S. aureus and C. albicans and demonstrates that SBC3 does not stimulate a non-specific immune response in larvae.

A free lunch? No cost for acquiring defensive plant pyrrolizidine alkaloids in a specialist arctiid moth (Utetheisa ornatrix).

Many herbivorous insects sequester defensive chemicals from their host plants. We tested sequestration fitness costs in the specialist moth Utetheisa ornatrix (Lepidoptera: Arctiidae). We added pyrrolizidine alkaloids (PAs) to an artificial diet at different concentrations. Of all the larval and adult fitness components measured, only development time was negatively affected by PA concentration. These results were repeated under stressful laboratory conditions. On the other hand, the amount of PAs sequestered greatly increased with the diet PA concentration. Absence of a detectable negative effect does not necessarily imply a lack of costs if all individuals express the biochemical machinery of detoxification and sequestration constitutively. Therefore, we used qPCR to show that expression of the gene used to detoxify PAs, pyrrolizidine-alkaloid-N-oxygenase (pno), increased 41-fold in our highest PA treatment. Nevertheless, fitness components were affected only slightly or not at all, suggesting that sequestration in this species does not incur a strong cost. The apparent lack of costs has important implications for our understanding of the evolution of ecological interactions; for example, it implies that selection by specialist herbivores may decrease the levels of certain chemical defences in plant populations.

Enhanced toxicity and induction of cytochrome P450s suggest a cost of "eavesdropping" in a multitrophic interaction.

The inducibility of cytochrome P450 monooxygenases (P450s) and other xenobiotic-metabolizing enzymes is thought to reflect material and energy costs of biosynthesis. Efforts to detect such costs of detoxification enzyme induction, however, have had mixed success. Although they are rarely considered, ecological costs of induction may be a more significant evolutionary constraint on herbivores than material and energy costs. Because some P450-mediated metabolic transformations are bioactivation reactions that increase, rather than reduce, toxicity, maintaining high levels of P450 activity places an organism at risk of greater mortality in the presence of compounds that are bioactivated. We show that P450 inducibility in the generalist moth Helicoverpa zea in response to plant signaling substances, an adaptive response in a ditrophic interaction between herbivore and plant, becomes detrimental in the presence of a third trophic association with a plant pathogen that produces aflatoxin, a toxin that can be bioactivated by P450s. Consumption of plant signaling molecules, such as methyl jasmonate (MeJA) and salicylic acid (SA) enhanced the toxicity of aflatoxin B1 (AFB1) to H. zea that resulted in substantially more damage to larval growth and development. Among the P450 transcripts already cloned from this organism, two in the CYP6B and CYP321A subfamilies are shown to be induced in response to MeJA and SA, suggesting that they may mediate some of the observed bioactivations.

Assessment of pesticide residues in army cutworm moths (Euxoa auxiliaris) from the Greater Yellowstone Ecosystem and their potential consequences to foraging grizzly bears (Ursus arctos horribilis).

During summer, a grizzly bear (Ursus arctos horribilis) in the Greater Yellowstone Ecosystem (GYE) (USA) can excavate and consume millions of army cutworm moths (Euxoa auxiliaris) (ACMs) that aggregate in high elevation talus. Grizzly bears in the GYE were listed as threatened by the US Fish and Wildlife Service in 1975 and were proposed for delisting in 2005. However, questions remain about key bear foods. For example, ACMs are agricultural pests and concern exists about whether they contain pesticides that could be toxic to bears. Consequently, we investigated whether ACMs contain and transport pesticides to bear foraging sites and, if so, whether these levels could be toxic to bears. In 1999 we collected and analyzed ACMs from six bear foraging sites. ACMs were screened for 32 pesticides with gas chromatography with electron capture detection (GC-ECD). Because gas chromatography with tandem mass spectrometry (GC-MS/MS) can be more sensitive than GC-ECD for certain pesticides, we revisited one site in 2001 and analyzed these ACMs with GC-MS/MS. ACMs contained trace or undetectable levels of pesticides in 1999 and 2001, respectively. Based on chemical levels in ACMs and numbers of ACMs a bear can consume, we calculated the potential of chemicals to reach physiological toxicity. These calculations indicate bears do not consume physiologically toxic levels of pesticides and allay concerns they are at risk from pesticides transported by ACMs. If chemical control of ACMs changes in the future, screening new ACM samples taken from bear foraging sites may be warranted.

[Food energy expenditure by the moth Lymantria dispar L. (Lepidoptera, Lymantriidae) at different stages of ontogenesis]. / Raskhodovanie énergii korma neparnym shelkopriadom Lymantria dispar L. (Lepidoptera, Lymantriidae) na raznykh étapakh ontogeneza.

Food energy expenditures by gypsy moth at different developmental stages under conditions of feeding on larch needles are described. The distribution of total food energy (%) for metabolism, the formation of exuvia (throughout ontogeny), and the formation of an adult insect is assessed. The proportion of food energy lost with excrements over the entire period of feeding is the same (58%) in females and males. The assimilated part of food energy is distributed differently: energy expenditures for the formation of exuvia and adult insects are greater in females than in males. The proportion of food energy expended for metabolism throughout the insect life span is approximately 30% in both females and males.