Pleiotropic immunoregulation by growth-blocking peptide in Ostrinia furnacalis.

Insects rely on their innate immune system to eliminate pathogenic microbes. As a system component, cytokines transmit intercellular signals to control immune responses. Growth-blocking peptide (GBP) is a member of the stress-responsive peptide family of cytokines found in several orders of insects, including Drosophila. However, the physiological role of GBP in defence against pathogens is not thoroughly understood. In this study, we explored the functions of GBP in a lepidopteran pest, Ostrinia furnacalis. Injection of recombinant O. furnacalis GBP (OfGBP) precursor (proGBP) and chemically synthesised GBP significantly induced the transcription of antimicrobial peptides (AMPs) and other immunity-related genes including immune deficiency (IMD) and Dorsal. The level of OfGBP mRNA was upregulated after bacterial infection. Knockdown of OfGBP expression led to a decrease in IMD, Relish, MyD88 and Dorsal mRNA levels. OfGBP induced phenoloxidase activity and affected hemocyte behaviours in O. furnacalis larvae. In summary, GBP is a potent cytokine, effectively regulating AMP synthesis, melanization response and cellular immunity to eliminate invading pathogens.

Identification and characteristic analysis of an extracellular signal-regulated kinase from Ostrinia furnacalis Guenée.

The extracellular signal-regulated kinase (ERK) pathway, a critical genetic determinant, controls diverse physiological functions, including innate immunity, development, and stress response. In the current study, a full-length cDNA (1592bp) encoding the ERK gene (OfERK) was cloned from Ostrinia furnacalis Guenée (GenBank accession number: MF797866). The open reading frame of the OfERK gene encoded 364 amino acids and shared 96.43%-98.08% amino acid identities with other insect mitogen-activated protein kinases. For spatiotemporal analysis of the expression pattern, OfERK exhibited a significant peak expression on the 3rd day of the pupa stage and showed the highest expression in hemocytes specifically. Indirect immunofluorescence assays and immuno-electron microscopy revealed a wide distribution of the OfERK protein in hemocytes and epidermis. Moreover, the results demonstrated that the Bt Cry1Ab-activated toxin significantly induces the expression of OfERK. Other genes related to immune response, development, and stress response exhibited dynamic changes in expression after Cry1Ab oral treatment. The expression of OfERK was downregulated through RNA interference, and the correlation of its expression with other related genes was verified using quantitative real-time polymerase chain reaction. Our study provides valuable insights into the regulatory mechanism of ERK in insects for future studies.

JAK/STAT signaling pathway and its regulation on insect immunity.

JAK/STAT signaling pathway is involved in important biological processes, such as cell growth, differentiation, apoptosis and immune regulation, and is one of the most important signal transduction pathways mediated by cytokines. JAK/STAT is a relatively conserved signaling pathway in insects, which is the main immune pathway together with Toll signaling pathway and Imd signaling pathway to resist the invasion of pathogens. It plays a vital role in insect immunity, hormone regulation and other physiological regulation processes. In this review, we summarize the cytokine receptor superfamily, JAKs family, STATs family, JAK/STAT signaling pathway and its mechanism of negative feedback regulation. We analyze the important functions and the latest research progress of JAK/STAT signaling pathway in insect infected by parasites, viruses and fungi. Finally, we propose the remaining problems in the study of JAK/STAT signaling pathway, so as to provide direction and reference for further research in this field.

Toxicity of Cry1-Class, Cry2Aa, and Vip3Aa19 Bt proteins and their interactions against yellow peach Moth, Conogethes punctiferalis (Guenée) (Lepidoptera: Crambidae).

Transgenic plants expressing insecticidal proteins from the Bacillus thuringiensis (Bt) have provided an effective way to control target pests. However, the toxicity of Bt proteins against yellow peach moth (YPM), Conogethes punctiferalis (Guenée), one of the most serious maize pests in China, has not received much study. Therefore, we performed diet-overlay bioassays to evaluate the insecticidal activities of Cry1Ab, Cry1Ac, Cry1Fa, Cry1Ah, Cry1Ie, Cry2Aa, and Vip3Aa19, as well as the interaction between Cry1-Class, Cry2Aa, and Vip3Aa19 against YPM. Results showed that the LC50 values ranged from 1.08 to 178.12 ng/cm2 (protein/diet). Among these proteins, Cry1Ab and Cry1Ac had lower LC50 values and LC90 values. In YPM bioassays, the combinations of Cry2Aa with Cry1Ac, Cry1Ie, and Cry1Ab showed antagonism while a mixture of Cry2Aa with Cry1Fa and Cry1Ah exhibited synergism. When Vip3Aa19 was combined with Cry proteins, all combinations interacted positively, with variation in synergistic factors (SF). Three ratios 1:1, 1:2, and 2:1 of Cry1Ah and Vip3Aa19 protein combination showed SF values of 5.20, 5.63, and 8.98, respectively. These findings can be applied in the establishment of new pyramided transgenic crops with suitable candidates as well as in resistance management strategies.

Transcriptome and Metabolome Analysis Reveals the Importance of Amino-Acid Metabolism in Spodoptera Frugiperda Exposed to Spinetoram.

Pests are inevitably exposed to sublethal and lethal doses in the agroecosystem following the application of pesticides indispensable to protect food sources. The effect of spinetoram on amino-acid metabolism of fall armyworm, Spodoptera frugiperda (J.E. Smith), was investigated, at the dose of LC10 and LC90, by transcriptome and LC-MS/MS analysis. Using statistics-based analysis of both POS and NEG mode, a total of 715,501 metabolites in S. frugiperda were significantly changed after spinetoram treatment. The enhancement of glucose metabolism provides energy support for detoxification in larvae. The decrease in valine and isoleucine is associated with an increase in leucine, without maintaining the conservation of citric acid in the larvae. The down-regulation of phenylalanine may retard the tricarboxylic acid cycle to produce GTP. The abundance of lysine was decreased in response to spinetoram exposure, which damages the nervous system of the larvae. The abundance of arginine increases and causes non-functional contraction of the insect's muscles, causing the larva to expend excess energy. Tryptophan provides an important substrate for eliminating ROS. The changes in glutamic acid, aspartic acid, and lysine cause damage to the nerve centers of the larvae. The results of transcriptome and LC-MS/MS analysis revealed the effects of pesticide exposure on amino-acid metabolism of S. frugiperda successfully and provide a new overview of the response of insect physio-biochemistry against pesticides.

Sublethal Effects of the Microbial-Derived Insecticide Spinetoram on the Growth and Fecundity of the Fall Armyworm (Lepidoptera: Noctuidae).

The fall armyworm, Spodoptera frugiperda (J. E. Smith), is one of the most important pests in tropical and subtropical regions of American. S. frugiperda was first detected in Southern China in January 2019, and then subsequently invaded in 26 provinces. Spinetoram widely used for pest management is recommended for S. frugiperda control. The sublethal effects of spinetoram on S. frugiperda were investigated in the present study. The toxicity of spinetoram against S. frugiperda larvae was determined after one oral dose of spinetoram at sublethal concentration. The results showed that spinetoram LC10 and LC30 were 0.011 and 0.044 mg/liter for the larvae, respectively. Spinetoram at sublethal concentration significantly increased developmental time but reduced larval body weight. In addition, spinetoram had a post-exposure effect on pupal weight, but not on pupal duration, pupation rate, emergence rate, eggs number, or adults longevity. In conclusion, the sublethal effects of spinetoram could negatively affect the growth and development of S. frugiperda that have important implications for pest management.

Comparison of larval performance and oviposition preference of Spodoptera frugiperda among three host plants: Potential risks to potato and tobacco crops.

Fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is a major polyphagous pest with the potential to seriously damage various crops. A better understanding of FAW's performance on different hosts may help to predict which plants will be attacked when the preferred host is absent, and facilitate the development of effective pest management practices. We compared the larval performance of FAW fed on maize with that of FAW fed on potato and tobacco, which are important crops in China, using an age-stage two-sex life table and adult female oviposition preference experiments. In cage experiments with potato, tobacco, or maize as the host, FAW reared on maize exhibited the strongest performance with shorter developmental time in the larval stage, longer longevity, and a higher reproductive rate in adults. Females oviposited on maize in preference to potato or tobacco. Compared with larvae fed on maize, those fed on potato and tobacco exhibited significantly lower survival, with only 31.61% and 8.13% developing to the adult stage, respectively. Several life table parameters, including the mean generation time (T), net reproductive rate (R0 ), finite rate of increase (λ), and intrinsic rate of natural increase (r) were negatively affected in FAW fed on potato and tobacco. Our results support the preference-performance hypothesis, that is, that herbivore females maximize fitness by choosing host plants associated with strong larval performance. Although larvae and adults performed poorly on potato and tobacco, their offspring will be capable of establishing populations on them, posing a potential threat to these crops in China.