The fall armyworm, Spodoptera frugiperda, has developed extremely high levels of resistance to chlorantraniliprole and other classes of insecticides in the field. As microRNAs (miRNAs) play important roles in various biological processes through gene regulation. we examined the miRNA profile of S. frugiperda in response to Chlorantraniliprole, Indoxacarb and Thiamethoxam. Transcriptome analysis showed significant changes in the abundance of some miRNAs after treatment of S. frugiperda larvae with LC20 concentrations of three insecticides. A total of 197 miRNAs were systematically identified from S. frugiperda, and 16, 9, 2 miRNAs were differentially expressed after treatments of three insecticides. Importantly, three miRNAs were significantly downregulated and three were upregulated by RT-qPCR after treatment the LC50 of three insecticides with S. frugiperda larvae. Microinjection of agomirs of these six miRNAs into S. frugiperda larvae resulted in significant changes in mortality rates when exposed to three insecticides. Additionally, we also screened potential target genes for some of differentially expressed miRNAs, which may play important roles in insecticide resistance development. These findings provide valuable insights into the molecular mechanisms of insecticide resistance and underscore the potential of miRNAs as targets for the development of novel pest control strategies in S. frugiperda.

Thiamethoxam (TMX) is a neonicotinoid insecticide. Residues of TMX have been detected in various crops. Although it has specific high toxicity to insects and is designed to exterminate them, the toxicity has also found in mammals recently. Differ from acetylcholine toxicity, TMX has peroxide toxicity in mammals. Matured oocytes have the capacity of fertilization, but oocytes own abundant mitochondria and its maturation is vulnerable to reactive oxygen species (ROS). Excessive production of reactive oxygen species (ROS) can override antioxidant defenses, produce oxidative stress and DNA damage that triggers apoptosis and necrosis in organisms. However little is known about the harm of ROS induced by TMX during oocytes maturation. Here, bovine germ-vesicle (GV) oocytes were cultured to metaphase of the second meiosis (MII) stage in vitro with or without TMX. During this process, oocytes were evaluated by various methods. Microscopic examination showed that 1.6 mM TMX significantly inhibited the maturation process in which oocytes were arrested before MI stage or between MI and MII stage. Correspond to this two periods, immunofluorescence staining and enzyme activity analysis showed that active CDC25 and CDC2 reduced in TMX group compared to control; time lapse and immunofluorescence staining gave results that Cyclin B could not be degraded, actin cap could not form, and Bub3 could not be removed from kinetochores. In addition, MII oocytes exposed to TMX showed disordered chromosomes and spindle. To study further, oocytes cultured for 24 h were analyzed. On the one hand, these oocytes in TMX group accumulated more ROS and produced significantly decreased mitochondrial membrane potential and increased apoptotic signal compared to control by methods of quantities for dichlorodihydrofluorescein diacetate (DCHFDA), 5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide and Annexin-V, but the level of γH2AX protein in TMX group did not decline significantly compared with control. On the another hand, these oocytes were activated to be parthenogenetic embryos and cultured. Assessment for embryo development showed decreased rates of cleavage, morula and blastocyst in TMX group compared to control in vitro. In conclusion, these results suggest that ROS induced by TMX results in dysfunction of mitochondria and apoptosis, which block bovine oocytes to MI stage, trap them at AI/TI stage and trigger disordered chromosomes and spindle at MII stage. Additionally, MII oocytes with poor qualities result from TMX lose abilities to cleavage and develop to be morulae and blastocysts.

This study was performed to investigate the effect of flonicamid and thiamethoxam treated at sublethal concentration (LC10, LC30) on development period, adult longevity and fecundity and the feeding behaviour of Myzus persicae adult. Developmental period of M. Persicae nymph took 5.9 days in LC10, and 6.1 days in LC30 in both insecticides, comparing with control (5.7 days), it showed longer than those of the control, but there was no significance. Adult longevity treated at LC10 and LC30 of flonicamid was showed 13.2 and 13.7 days, respectively, and LC10 of thiamethoxam was examined as 14.7 days, it showed longer than control of 11.6 days. Mean daily fecundity exhibited higher in LC10 (3.1) and LC30 (3.1) of flonicamid than that of control (2.5), but thiamethoxam are not. Total fecundity exhibited higher in LC10 (41.8) and LC30 (43.0) of flonicamid, in LC10 (42.1) of thiamethoxam than that of control (29.5). Feeding behavior was examined using EPG (electrical penetration graph). EPG data indicated that flonicamid and thiamethoxam increased the duration of non-probing periods and decreased the duration of phloem ingestion.

To investigate resistance of Myzus persicae, which is resistant to etofenprox belonging to pyrethriods, to other insecticides, fenpropathrin and thiamethoxam, mortality and LC₅₀ values for these two insecticides against the etofenprox-resistant and -susceptible populations were obtained and EPG analysis for feeding behaviors of thiamethoxam or fenpropathrin treated and non-treated M. persicae was conducted. For fenpropathrin, mortality after 48 h treatment was 28 and 29%, respectively for the resistant and susceptible population. The LC₅₀ value was 193.15 and 93.46 ppm, respectively. For thiamethoxam, mortality after 48 h treatment was 87 and 57%, respectively for the resistant and susceptible population. The LC₅₀ value was 3.17 and 30.34 ppm, respectively. There was no significant difference in feeding behavior between fenprorpathrin treated and non-treated M. persicae. They showed a continuous feeding pattern. However, contact signal frequency increased in the thiamethoxam treated M. persicae 1 hour after treatment and significant difference was shown.