Juvenile hormone induces reproduction via miR-1175-3p in the red imported fire ant, Solenopsis invicta.

Juvenile hormone (JH) acts in the regulation of caste differentiation between queens and workers (i.e., with or without reproductive capacity) during vitellin synthesis and oogenesis in social insects. However, the regulatory mechanisms have not yet been elucidated. Here, we identified a highly expressed microRNA (miRNA), miR-1175-3p, in the red imported fire ant, Solenopsis invicta. We found that miR-1175-3p is prominently present in the fat bodies and ovaries of workers. Furthermore, miR-1175-3p interacts with its target gene, broad-complex core (Br-C), in the fat bodies. By utilizing miR-1175-3p agomir, we successfully suppressed the expression of the Br-C protein in queens, resulting in reduced vitellogenin expression, fewer eggs, and poorly developed ovaries. Conversely, decreasing miR-1175-3p levels led to the increased expression of Br-C and vitellogenin in workers, triggering the "re-development" of the ovaries. Moreover, when queens were fed with JH, the expression of miR-1175-3p decreased, whereas the expression of vitellogenin-2 and vitellogenin-3 increased. Notably, the suppression of fertility in queens caused by treatment with agomir miR-1175-3p was completely rescued by the increased vitellogenin expression induced by being fed with JH. These results suggest the critical role of miR-1175-3p in JH-regulated reproduction, shedding light on the molecular mechanism underlying miRNA-mediated fecundity in social insects and providing a novel strategy for managing S. invicta.

Labor division of worker ants can be controlled by insulin synthesis targeted through miR-279c-5p in Solenopsis invicta (Hymenoptera: Formicidae).

BACKGROUND: In social insects, the labor division of workers is ubiquitous and controlled by genetic and environmental factors. However, how they modulate this coordinately remains poorly understood. RESULTS: We report miR-279c-5p participation in insulin synthesis and behavioral transition by negatively regulating Rab8A in Solenopsis invicta. Eusocial specific miR-279c-5p is age-associated and highly expressed in nurse workers, and localized in the cytoplasm of neurons, where it is partly co-localized with its target, Rab8A. We determined that miR-279c-5p agomir suppressed Rab8A expression in forager workers, consequently decreasing insulin content, resulting in the behavioral shift to 'nurse-like' behaviors, while the decrease in miR-279c-5p increased Rab8A expression and increased insulin content in nurse workers, leading to the behavioral shift to 'foraging-like' behaviors. Moreover, insulin could rescue the 'foraging behavior' induced by feeding miR-279c-5p to nurse workers. The overexpression and suppression of miR-279c-5p in vivo caused an obvious behavioral transition between foragers and nurses, and insulin synthesis was affected by miR-279c-5p by regulating the direct target Rab8A. CONCLUSION: We first report that miR-279c-5p is a novel regulator that promotes labor division by negatively regulating the target gene Rab8A by controlling insulin production in ants. This miRNA-mediated mechanism is significant for understanding the behavioral plasticity of social insects between complex factors and potentially provides new targets for controlling red imported fire ants. © 2023 Society of Chemical Industry.

Osa-miR162a Enhances the Resistance to the Brown Planthopper via α-Linolenic Acid Metabolism in Rice (Oryza sativa).

The brown planthopper (BPH) is the most serious pest causing yield losses in rice. MicroRNAs (miRNAs) are emerging as key modulators of plant-pest interactions. In the study, we found that osa-miR162a is induced in response to BPH attack in the seedling stage and tunes rice resistance to the BPH via the α-linolenic acid metabolism pathway as indicated by gas chromatography/liquid chromatography-mass spectrometry analysis. Overexpression of osa-miR162a inhibited the development and growth of the BPH and simultaneously reduced the release of 3-hexenal and 3-hexen-1-ol to block host recognition in the BPH. Moreover, knockdown of OsDCL1, which is targeted by osa-miR162a, inhibited α-linolenic acid metabolism to enhance the resistance to the BPH, which was similar to that in miR162a-overexpressing plants. Our study revealed a novel defense mechanism mediated by plant miRNAs developed during the long-term evolution of plant-host interaction, provided new ideas for the identification of rice resistance resources, and promoted a better understanding of pest control.

Halloysite Nanotube-Based Pesticide Formulations with Enhanced Rain Erosion Resistance, Foliar Adhesion, and Insecticidal Effect.

The incorporation of green and sustainable nanomaterials in pesticide formation is an effective method to lower the use of conventional pesticides without adverse effects on productivity. Here pesticide Pickering emulsions stabilized by halloysite nanotubes (HNTs) were developed for low cost, less environmental pollution, low toxic effects, and better emulsion stability. HNTs were added to chlorantraniliprole (CAP) emulsions, and good stability was exhibited due to the adsorption and aggregation of HNTs at the interface of CAP oil droplets, forming a three-dimensional network structure that prevented the emulsion from aggregation. In addition, Spodoptera frugiperda was used as a pest model and corn was used as a plant model to explore the washout resistance, insecticidal effect, and biological safety of HNTs-CAP emulsion. After spraying emulsion on corn leaves and washing for 10 min, the HNTs-CAP emulsion (5 wt % HNTs) pesticide residue rate was 2.7 times that of pristine CAP emulsion. When the HNT dispersion concentration was 2 wt %, the larva mortality was 83%, which was 1.5 times that of the CAP emulsion group. These results demonstrated that HNTs-CAP emulsion showed good foliar adhesion, rainfall resistance, and insecticidal effect. The tubular clay-based nanopesticide formulations show potential applications in the control of crop pests with modern agriculture technology.

Role of the foraging gene in worker behavioral transition in the red imported fire ant, Solenopsis invicta (Hymenoptera: Formicidae).

BACKGROUND: Worker division of labor is predominant in social insects. The foraging (for) gene, which encodes cGMP-dependent protein kinase (PKG), has been implicated in the regulation of behavioral transitions in honeybees, but information regarding its function in other social insects is scarce. RESULTS: We investigated the role of the for (Sifor) gene in the red imported fire ant, Solenopsis invicta, and found that Sifor and PKG exhibited different expression patterns in different castes, body sizes, ages and tissues of fire ants, especially in foragers and nurses. Foragers displayed greater locomotor activity but showed no preference for larval or adult odors, whereas nurses showed lesser locomotor activity but had a strong preference for larval odors. We found that the expression of Sifor was significantly higher in the heads of foragers (compared to nurses). RNA interference-mediated Sifor knockdown in foraging workers induced behavioral transition of foragers toward the nurse phenotype characterized by reduced locomotor activity and a stronger preference for larval odors. By contrast, treating nurses with 8-Br-cGMP, an activator of PKG, resulted in behavioral transition toward the forager phenotype characterized by higher locomotor activity but reduced preference for larval odors. CONCLUSION: Our results suggest that Sifor plays a critical role in the behavioral transition between foragers and nurses of workers, which may be a promising target for RNAi-based management of worker caste organization in S. invicta. © 2022 Society of Chemical Industry.

Preference and effect of gustatory sense on sugar-feeding of fire ants.

BACKGROUND: The red imported fire ant is one of the notorious species of ants all over the world. Sugar is one of the most important components of food and necessary for the survival of ants. Because more than 70% food of fire ants is honeydew produced by Homopteran insects such as aphids and scales. METHODOLOGY: It is well known that beetles, flies, and honey bees can recognize the sugar taste through their legs and antennae, but in the case of fire ants, no records regarding gustatory sense were published. In the current study, considering the importance of sugar bait, we investigated the gustatory sense of the fire ant workers to sucrose via behavioral sequence and gustatory behavior. First, the feeding sequence (ethogram) of the fire ant workers on most preferred sugar (sucrose) solution was observed and categorized. Secondly, the gustatory behavior of treated fire ant workers (without flagellum and foreleg tarsi treated with HCL solution) was observed on the sucrose solution. In addition, using scanning electron microscopy (SEM) techniques, we identified the possible porous sensilla types on antenna flagellum and foreleg tarsi of fire ant workers. RESULTS: Based on the results of feeding sequence, foreleg tarsi of workers were the main body appendages in the detection of the sucrose droplet as compared to antennae flagellum and palps. Feeding time of treated workers with HCL solution was significantly decreased on sucrose solution as compared to those workers having no flagellum. While both types of treated workers have less feeding time in comparison to normal workers. Based on the results of feeding sequence analysis and feeding time, it is indicating that the foreleg tarsi of workers play a more important role in the detection of sucrose solution as compared to antennae flagellum. Based on the SEM results, sensilla chaetic, trichoid II, and basiconic I and II have a clear pore at their tip. This study provides a substantial basis for elucidating the gustatory function of antennal and tarsal sensilla on appendages of fire ant workers to sugars and further baits improvement for the management of fire ants.

Thermally Induced Actinidine Production in Biological Samples.

Actinidine, a methylcyclopentane monoterpenoid pyridine alkaloid, has been found in many iridoid-rich plants and insect species. In a recent research on a well-known actinidine- and iridoid-producing ant species, Tapinoma melanocephalum (Fabricius) (Hymenoptera: Formicidae), no actinidine was detected in its hexane extracts by gas chromatography-mass spectrometry analysis using a common sample injection method, but a significant amount of actinidine was detected when a solid injection technique with a thermal separation probe was used. This result led us to hypothesize that heat can induce the production of actinidine in iridoid-rich organisms. To test our hypothesis, the occurrence of actinidine was investigated in four iridoid-rich organisms under different sample preparation temperatures, including two ant species, T. melanocephalum and Iridomyrmex anceps Roger (Hymenoptera: Formicidae), and two plant species, Actinidia polygama Maxim (Ericales: Actinidiaceae) and Nepeta cataria L. (Lamiales: Lamiaceae). Within a temperature range of 50, 100, 150, 200, and 250 °C, no actinidine was detected at 50 °C, but it appeared at temperatures above 100 °C for all four species. A positive relationship was observed between the heating temperature and actinidine production. The results indicate that actinidine could be generated at high temperatures. We also found that the presence of methylcyclopentane monoterpenoid iridoids (iridodials and nepetalactone) was needed for thermally induced actinidine production in all tested samples. These results suggest that the presence of actinidine in iridoid-rich plants and ants might be a consequence of using high temperatures during sample preparation.

Volatile Terpenes and Terpenoids from Workers and Queens of Monomorium chinense (Hymenoptera: Formicidae).

Twenty-one volatile terpenes and terpenoids were found in Monomorium chinense Santschi (Hymenoptera: Formicidae), a native Chinese ant, by using headspace solid-phase microextraction (HS-SPME) coupled with gas-phase chromatography and mass spectrometry (GC-MS), which makes this ant one of the most prolific terpene producers in insect. A sesquiterpene with unknown structure (terpene 1) was the main terpene in workers and neocembrene in queens. Terpenes and terpenoids were detected in poison, Dufour's and mandibular glands of both workers and queens. Worker ants raised on a terpene-free diet showed the same terpene profile as ants collected in the field, indicating that de novo terpene and terpenoid synthesis occurs in M. chinense.

Gland Origin and Electroantennogram Activity of Volatile Compounds in Ghost Ants, Tapinoma melanocephalum (Hymenoptera: Formicidae) and Behavioral Response to (Z)-9-Nonadecene.

Volatile compounds in Tapinoma melanocephalum (Fabricius) (Hymenoptera: Formicidae, Dolichoderinae) workers were analyzed using gas chromatography-mass spectrometry and electroantennogram responses of workers to these compounds were investigated using coupled gas chromatography-electroantennographic detection. Among 18 detected compounds, six elicited electroantennogram response, including 6-methyl-5-hepten-2-one, 4 iridodials, and (Z)-9-nonadecene. (Z)-9-Nonadecene and iridodials were identified the first time in T. melanocephalum. (Z)-9-Nonadecene is a major component in mandibular glands and iridodials are produced in pygidial glands. In contrast to previous report, actinidine was not found in pygidial glands. Behavioral response of workers to the synthetic (Z)-9-nonadecene was investigated. (Z)-9-nonadecene is an attractant to T. melanocephalum. It also affected their locomotion patterns.

Effects of Insecticides on Oviposition and Host Discrimination Behavior in Trichogramma chilonis (Hymenoptera: Trichogrammatidae).

Trichogramma chilonis Ishii is an important natural enemy of many lepidopterous pests on various crops. Effects of two conventional insecticides on oviposition and host discrimination behavior in T. chilonis were evaluated in the laboratory through video tracking. During the oviposition period, when the number of host eggs was limited (only one host egg), females exposed to beta-cypermethrin LC1 or LC20 exhibited significantly higher feeding and re-drilling rate and significantly longer post-oviposition duration than control females. Spinosad LC20 treatment not only decreased the wasp oviposition rate but also significantly extended the oviposition duration. When the number of host eggs was six, similarly, females exposed to spinosad LC20 exhibited a significantly lower oviposition rate (79.2%) than control ones (100%). In the host discrimination experiment, females exposed to LC20 of both tested insecticides (beta-cypermethrin and spinosad) spent significantly more time on the extra-patch area. The females that survived spinosad LC20 could not discriminate between unparasitized and parasitized host eggs. Our study suggests that even the LC1 and LC20 of the tested insecticides had negative effects on the oviposition and host discrimination behavior of T. chilonis Thus, the use of these two insecticides should be carefully evaluated.