Endophytic Fungi-Mediated Defense Signaling in Maize: Unraveling the Role of WRKY36 in Regulating Immunity against Spodoptera frugiperda.

Seed priming with beneficial endophytic fungi is an emerging sustainable strategy for enhancing plant resistance against insect pests. This study examined the effects of Beauvaria bassiana Bb20091317 and Metarhizium rileyi MrCDTLJ1 fungal colonization on maize growth, defence signalling, benzoxazinoid levels and gene expression. The colonization did not adversely affect plant growth but reduced larval weights of Spodoptera frugiperda. Maize leaves treated with M. rileyi exhibited higher levels of jasmonic acid, jasmonoyl-Isoleucine, salicylic acid, and indole acetic acid compared to control. B. bassiana and M. rileyi accelerated phytohormone increase upon S. frugiperda herbivory. Gene expression analysis revealed modulation of benzoxazinoid biosynthesis genes. We further elucidated the immune regulatory role of the transcription factor zmWRKY36 using virus-induced gene silencing (VIGS) in maize. zmWRKY36 positively regulates maize immunity against S. frugiperda, likely by interacting with defense-related proteins. Transient overexpression of zmWRKY36 in tobacco-induced cell death, while silencing in maize reduced chitin-triggered reactive oxygen species burst, confirming its immune function. Overall, B. bassiana and M. rileyi successfully colonized maize, impacting larval growth, defense signalling, and zmWRKY36-mediated resistance. This sheds light on maize-endophyte-insect interactions for sustainable plant protection.

Antagonistic and additive effect when combining biopesticides against the fall armyworm, Spodoptera frugiperda.

Fall armyworm, Spodoptera frugiperda (FAW) is a cosmopolitan crop pest species that has recently become established in sub-Saharan Africa and Southeast Asia. Current FAW control is almost entirely dependent on synthetic pesticides. Biopesticides offer a more sustainable alternative but have limitations. For example, pyrethrum is an effective botanical insecticide with low mammalian toxicity but is highly UV labile, resulting in a rapid loss of efficacy in the field. Beauveria bassiana is an entomopathogenic fungus that is more persistent, but there is a time lag of several days before it causes insect mortality and leads to effective control. The combination of these biopesticides could mitigate their drawbacks for FAW control. Here we evaluated the efficacy of pyrethrum and B. bassiana as individual treatments and in combination against 3rd instar FAW. Four different combinations of these two biopesticides were tested, resulting in an antagonistic relationship at the lowest concentrations of B. bassiana and pyrethrum (1 × 104 conidia mL-1 with 25 ppm) and an additive effect for the other 3 combined treatments (1 × 104 conidia mL-1 with 100 ppm and 1 × 105 conidia mL-1 with 25 ppm and 100 ppm pyrethrum). Additionally, a delay in efficacy from B. bassiana was observed when combined with pyrethrum as well as a general inhibition of growth on agar plates. These results appear to show that this particular combination of biopesticides is not universally beneficial or detrimental to pest control strategies and is dependent on the doses of each biopesticide applied. However, the additive effect shown here at specific concentrations does indicate that combining biopesticides could help overcome the challenges of persistence seen in botanical pesticides and the slow establishment of EPF, with the potential to improve effectiveness of biopesticides for IPM.

SfMBP: A novel microbial binding protein and pattern recognition receptor in the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae).

The fall armyworm, Spodoptera frugiperda, poses a significant threat as a highly destructive agricultural pest in many countries. Understanding the complex interplay between the insect immune system and entomopathogens is critical for optimizing biopesticide efficacy. In this study, we identified a novel microbial binding protein, SfMBP, in S. frugiperda. However, the specific role of SfMBP in the immune response of S. frugiperda remains elusive. Encoded by the LOC118269163 gene, SfMBP shows significant induction in S. frugiperda larvae infected with the entomopathogen Beauveria bassiana. Consisting of 115 amino acids with a signal peptide, an N-terminal flexible region and a C-terminal ß-sheet, SfMBP lacks any known functional domains. It is expressed predominantly during early larval stages and in the larval epidermis. Notably, SfMBP is significantly induced in larvae infected with bacteria and fungi and in SF9 cells stimulated by peptidoglycan. While recombinant SfMBP (rSfMBP) does not inhibit bacterial growth, it demonstrates binding capabilities to bacteria, fungal spores, peptidoglycan, lipopolysaccharides, and polysaccharides. This binding is inhibited by monosaccharides and EDTA. Molecular docking reveals potential Zn2+-interacting residues and three cavities. Furthermore, rSfMBP induces bacterial agglutination in the presence of Zn2+. It also binds to insect hemocytes and SF9 cells, enhancing phagocytosis and agglutination responses. Injection of rSfMBP increased the survival of S. frugiperda larvae infected with B. bassiana, whereas blocking SfMBP with the antibody decreased survival. These results suggest that SfMBP acts as a pattern recognition receptor that enhances pathogen recognition and cellular immune responses. Consequently, this study provides valuable insights for the development of pest control measures.

Cowpea volatiles induced by beet armyworm or fall armyworm differentially prime maize plants.

Exposure to herbivore-induced plant volatiles (HIPVs) is known to enhance the defense responses in plants. This so-called priming effect has only been marginally studied in intercropping systems. We tested whether HIPVs from cowpea, which often serves as an intercrop alongside maize, can prime herbivore-induced volatile emissions in maize. Conventional volatile collection assays and real-time mass spectrometry revealed that maize plants that were exposed to HIPVs from cowpea infested with Spodoptera exigua caterpillars emitted more than control plants when they themselves were subsequently damaged by the same pest. The enhanced emission was only evident on the first day after infestation. Maize plants that were exposed to HIPVs from cowpea infested by S. frugiperda larvae showed no priming effect and released considerably less upon S. frugiperda infestation than upon S. exigua infestation. The latter may be explained by the fact that S. frugiperda is particularly well adapted to feed on maize and is known to suppress maize HIPV emissions. Our results imply that HIPVs from cowpea, depending on the inducing insect herbivore, may strongly prime maize plants. This deserves further investigation, also in other intercropping systems, as it can have important consequences for tritrophic interactions and crop protection.

Ionotropic Receptor IR75q.2 Mediates Avoidance Reaction to Nonanoic Acid in the Fall Armyworm Spodoptera frugiperda (Lepidoptera, Noctuidae).

Ionotropic receptors (IRs) play an important role in olfaction, but little is known in nondrosophila insects. Here, we report in vitro and in vivo functional characterization of IR75q.2 in the invasive moth pest Spodoptera frugiperda. First, 13 IRs (including four coreceptor IRs) were found specifically or highly expressed in adult antennae. Second, these IRs were tested for responding profiles to 59 odorants using the Xenopus oocyte expression system, showing that only SfruIR75q.2 responded to 8-10C fatty acids and their corresponding aldehydes, with SfruIR8a as the only coreceptor. Third, the three acids (especially nonanoic acid) showed repellent effects on moth's behavior and oviposition, but the repellence significantly reduced to the insects with IR75q.2 knockout by CRISPR/Cas9. Taken together, our study reveals the function of SfruIR75q.2 in perception of acid and aldehyde odorants and provides the first in vivo evidence for olfactory function of an odor-specific IR in Lepidoptera.

Transcriptome Dynamics of Brassica juncea Leaves in Response to Omnivorous Beet Armyworm (Spodoptera exigua, Hübner).

Cruciferous plants manufacture glucosinolates (GSLs) as special and important defense compounds against insects. However, how insect feeding induces glucosinolates in Brassica to mediate insect resistance, and how plants regulate the strength of anti-insect defense response during insect feeding, remains unclear. Here, mustard (Brassica juncea), a widely cultivated Brassica plant, and beet armyworm (Spodoptera exigua), an economically important polyphagous pest of many crops, were used to analyze the changes in GSLs and transcriptome of Brassica during insect feeding, thereby revealing the plant-insect interaction in Brassica plants. The results showed that the content of GSLs began to significantly increase after 48 h of herbivory by S. exigua, with sinigrin as the main component. Transcriptome analysis showed that a total of 8940 DEGs were identified in mustard challenged with beet armyworm larvae. The functional enrichment results revealed that the pathways related to the biosynthesis of glucosinolate and jasmonic acid were significantly enriched by upregulated DEGs, suggesting that mustard might provide a defense against herbivory by inducing JA biosynthesis and then promoting GSL accumulation. Surprisingly, genes regulating JA catabolism and inactivation were also activated, and both JA signaling repressors (JAZs and JAMs) and activators (MYCs and NACs) were upregulated during herbivory. Taken together, our results indicate that the accumulation of GSLs regulated by JA signaling, and the regulation of active and inactive JA compound conversion, as well as the activation of JA signaling repressors and activators, collectively control the anti-insect defense response and avoid over-stunted growth in mustard during insect feeding.

Bionomic responses of Spodoptera frugiperda (J. E. Smith) to lethal and sublethal concentrations of selected insecticides.

Since 2016, the invasive insect Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) from the Americas has made maize production unattainable without pesticides in parts of Sub-Saharan Africa and Asia. To counteract this pest, farmers often resort to the use hazardous pesticides. This study aimed to investigate botanicals, microbials, and semi-synthetic insecticides in Ghana for pest control without harming local ecosystems. Under laboratory and on-station conditions, the present study evaluated the acute and sublethal responses of S. frugiperda to: (i) Pieris rapae Granulovirus (PrGV) + Bacillus thuringiensis sub sp. kurstaki (Btk) 5 WP, (ii) Btk + monosultap 55 WP, (iii) ethyl palmitate 5 SC, (iv) azadirachtin 0.3 SC, (v) acetamiprid (20 g/l) + λ-cyhalothrin (15 g/l) 35 EC, (vi) acetamiprid (30 g/l) + indoxacarb (16 g/l) 46 EC, and (vii) emamectin benzoate 1.9 EC. The results showed that at 96 hours post-exposure emamectin benzoate-based formulation has the highest acute larvicidal effect with lower LC50 values of 0.019 mL/L. However, the results suggested strong sublethal effects of PrGV + Btk, azadirachtin, and ethyl palmitate on the bionomics of S. frugiperda. Two seasons on-station experiments, showed that the semi-synthetic emamectin benzoate and the bioinsecticide PrGV + Btk are good candidates for managing S. frugiperda. The promising efficacy of emamectin benzoate and PrGV + Btk on the bionomics of S. frugiperda in the laboratory and on-station demonstrated that they are viable options for managing this pest.

DIMBOA-induced gene expression, activity profiles of detoxification enzymes, multi-resistance mechanisms, and increased resistance to indoxacarb in tobacco cutworm, Spodoptera litura (Fabricius).

Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) is one of the most destructive insect pests owned strong resistance to different insecticides. Indoxacarb as a novel oxadiazine insecticide becomes the main pesticide against S. litura. DIMBOA [2,4-dihydroxy-7-methoxy-2 H-1,4-benz-oxazin-3(4 H)-one] is involved in important chemical defense processes in corn plants. However, the insects' adaptation mechanism to insecticides when exposed to defensive allelochemicals in their host plants remains unclear. Here, we assessed multi-resistance, and resistance mechanisms based on S. litura life history traits. After 18 generations of selection, indoxacarb resistance was increased by 61.95-fold (Ind-Sel) and 86.06-fold (Dim-Sel) as compared to the Lab-Sus. Also, DIMBOA-pretreated larvae developed high resistance to beta-cypermethrin, chlorpyrifos, phoxim, chlorantraniliprole, and emamectin benzoate. Meanwhile, indoxacarb (LC50) was applied to detect its impact on thirty-eight detoxification-related genes expression. The transcripts of SlituCOE073, SlituCOE009, SlituCOE074, and SlituCOE111 as well as SlGSTs5, SlGSTu1, and SlGSTe13 were considerably raised in the Ind-Sel strain. Among the twenty-three P450s, CYP6AE68, CYP321B1, CYP6B50, CYP9A39, CYP4L10, and CYP4S9v1 transcripts denoted significantly higher levels in the Ind-Sel strain, suggesting that CarEs, GSTs and P450s genes may be engaged in indoxacarb resistance. These outcomes further highlighted the importance of detoxification enzymes for S. litura gene expression and their role in responses to insecticides and pest management approaches.

Modeling the potential global distribution of the Egyptian cotton leafworm, Spodoptera littoralis under climate change.

The Egyptian cotton leafworm, Spodoptera littoralis is a highly invasive insect pest that causes extensive damage to many of the primary food crops. Considering the recent challenges facing global food production including climate change, knowledge about the invasive potential of this pest is essential. In this study, the maximum entropy model (MaxEnt) was used to predict the current global spatial distribution of the pest and the future distribution using two representative concentration pathways (RCPs) 2.6 and 8.5 in 2050 and 2070. High AUC and TSS values indicated model accuracy and high performance. Response curves showed that the optimal temperature for the S. littoralis is between 10 and 28 °C. The pest is currently found in Africa and is widely distributed across the Middle East and throughout Southern Europe. MaxEnt results revealed that the insect will shift towards Northern Europe and the Americas. Further, China was seen to have a suitable climate. We also extrapolated the impact of these results on major producing countries and how this affects trade flow, which help decision makers to take the invasiveness of such destructive pest into their account.

Adaptation by death? Cell death-based tolerance to cadmium in 150-generation exposure of Spodoptera exiqua Hübner (Lepidoptera: Noctuidae).

Mechanisms, including autophagy and apoptosis, which serve to regulate and ensure proper organism functions under optimal conditions, play additional defensive roles under environmental pressure. The aim of this study was to test the following hypotheses: (i) elevated autophagy and apoptosis intensity levels, as defensive processes in response to contact with cadmium, are maintained for a limited number of generations and (ii) the number of generations after which levels of cell death processes reach the reference level depends on selective pressure. Cell death processes were assessed by light and transmission electron microscopy, terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL), and cytometric analyses. Model insects (Spodoptera exiqua, Hübner, 1808) were orally exposed to various concentrations of cadmium for 18 generations and compared with reference strains exposed to cadmium or not (control) for over 150 generations. Elevated programmed cell death intensity levels decreased after several generations, indicating tolerance of individuals to cadmium in the diet and verifying the first hypothesis; however, testing the second hypothesis indicated that the number of generations depended not only on pressure intensity, but also on cell death type, since levels of autophagy remained increased for a minimum of 12 generations.

Physiological Function of RDL1 and RDL2 Subunits of the Ionotropic GABA Receptor in the Spodoptera litura with the CRISPR/Cas9 System In Vivo.

In insect ionotropic γ-aminobutyric acid receptor (iGABAR) subunits, only resistance to dieldrin (RDL) can be individually and functionally expressed in vitro. In lepidopteran, two to three RDL subtypes are identified; however, their physiological roles have not been distinguished in vivo. In this study, SlRdl1 and SlRdl2 of S. litura were individually knocked out using CRISPR/Cas9, respectively. The mortality and larval and pupal duration of KOSlRdl1 and KOSlRdl2 were increased. The flight time and distance were increased by 43.30%-80.66% and 58.96%-198.22%, respectively, in KOSlRdl1. The GABA-induced current was significantly decreased by 53.57%-74.28% and 46.91%-63.34% in the ventral nerve cord, and the GABA titer was significantly reduced by 17.65%-28.05% and 19.85%-42.46% in KOSlRdl1 and KOSlRdl2, respectively. In conclusion, SlRdl1 and SlRdl2 are necessary for the transmission of GABA-induced neural signals; however, only SlRdl1 could regulate the flight capability of S. litura. Our results provided a new avenue to study lepidopteran iGABARs.

RNAi-mediated silencing of SlitPer disrupts sex pheromone communication behavior in Spodoptera litura.

BACKGROUND: The 24-h circadian rhythm is considered crucial for insect sexual communication. However, its molecular mechanisms and signaling pathways, particularly the roles of the clock gene period (Per), remain largely unclear. The sex pheromone communication behavior of Spodoptera litura displays typical circadian rhythm characteristics. Thus, it represents an excellent model for functional analyses of the clock gene Per. RESULTS: In this study, we investigated the potential roles of SlitPer in regulating sex pheromone communication in S. litura using RNA interference, quantitative real-time polymerase chain reactions (qPCR), gas chromatography, and behavioral assays. The qPCR results showed that the expression levels of SlitPer and two desaturase genes (SlitDes5 and SlitDes11) in the siPer group differed significantly at most time points from those in the siNC group. Dynamic variation in the three major sex pheromone titers and calling behavior of S. litura females in the siPer group was disordered. In addition, the mating rates of siPer S. litura females decreased significantly by 33.33%. Oviposition by mated siPer females was substantially reduced by 84.84%. CONCLUSION: These findings provide a fundamental basis for elucidating the molecular mechanism by which Per regulates sex pheromone communication behavior in lepidopteran species. © 2023 Society of Chemical Industry.

Transgenic Drosophila to Functionally Validate Fall Armyworm ABCC2 Mutations Conferring Bt Resistance.

The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith; Lepidoptera: Noctuidae) is an invasive agricultural pest with a global distribution, causing major crop losses annually. Its control strategies largely rely on chemical insecticides and transgenic crops expressing Bacillus thuringiensis insecticidal proteins (Cry and Vip toxins); however, the development of high resistance poses a significant issue. The ATP-binding cassette transporter C2 (ABCC2) has been linked to Cry toxin pore formation, acting as a receptor of some Cry toxins. Recently detected mutations in the SfABCC2 gene in extracellular loop 4 (ECL4) have been associated with Bt toxin resistance in FAW. In the present study, we expressed the SfABCC2 gene in Drosophila melanogaster, a species normally unaffected by the Bt toxins. We demonstrate that susceptibility can be introduced by the ectopic and tissue-specific expression of wildtype SfABCC2. Next, we introduced mutations into ECL4-both individually and in combination-that have been recently described in Brazilian FAW and functionally validated by toxicity bioassays against the foliar Bt product Xentari. Our results provide an efficient demonstration of the suitability of transgenic Drosophila for validating FAW ABCC2 resistance mutations in ECL4 against Bt toxins, and potential cross-resistance issues between closely related proteins that use ABCC2.

Population density of the fall armyworm, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) and its response to some ecological phenomena in maize crop.

The fall armyworm [FAW; Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae)], is considered a serious invasive pest that poses a serious threat to world food security. It can completely devastate a whole country's cereal crops. Therefore, the present work is the 1st field trial in Egypt to elucidate some ecological aspects of S. frugiperda on maize plants (Single-Hybrid 168 Yellow Corn cultivar) at Esna district, Luxor Governorate, Egypt, throughout two sequential growing seasons of maize (2021-2022). Three insect parameters were used, i.e., egg masses, number of larvae, and percentage of damaged corn plants. Effects of certain climatic conditions and corn plant ages on S. frugiperda seasonal activity and damaged plants percentage were also estimated. S. frugiperda population initiated to attack maize plants from the 3rd week of June until the harvest in every season. S. frugiperda had two seasonal activity peaks in terms of egg masses numbers and three peaks regarding the larval population density/season. Its damage percentage increased with increasing plant age weekly. The mean of S. frugiperda egg masses were 2.83 ± 0.40 and 2.96 ± 0.45 mass /10 corn plants in 2021 and 2022, respectively. While, the overall mean larval populations were 13.41 ± 0.52 and 13.03 ± 0.46 larvae/10 plants, during the two growing seasons, respectively. Corn plant damage percentages reached 68.54 ± 2.71 and 60.42 ± 2.92% in 2021 and 2022, respectively. The combined effects of both the weather conditions and maize plant ages were highly significant on egg masses, larvae population density, and damage percentage, and varied from one season to another. The mean daily maximum temperature was the most effective variable on egg masses and the larval population. Maize plant age had a clear effect on the damage percentage caused by the larvae in the field during the two years of the study. The dramatic spread of FAW and the consequent damage (that appeared in different countries of America, Africa, and Asia) mean that different management approaches must be sought for the small and large-scale producers by using available technologies for smallholder farmers will eliminate pest damage without access to reach to an economic Injury level. This information may assist the decision maker when planning the S. frugiperda IPM program for maize plants and its surveillance.

Feeding responses and digestive function of Spodoptera littoralis (Boisd) on various leafy vegetables exhibit possible tolerance traits.

Spodoptera littoralis is a highly polyphagous pest that attacks numerous important crops in the world and causes substantial economic losses to agricultural production. In the present study, the effects of different leafy vegetables, including Purslane, Chives, Parsley, Basil, Dill, Coriander, and Mint, were investigated on feeding responses and enzymatic activities of S. littoralis under laboratory conditions. Furthermore, the total contents of the three major secondary metabolites (phenolics, anthocyanins, and flavonoids) in the studied vegetables were determined. Our findings showed that the lowest and the highest approximate digestibility were on Basil and Purslane, respectively. The highest values of efficiency of conversion of ingested and digested food were achieved in larvae fed on Chives and Coriander, respectively, whereas the lowest values were recorded after feeding on Purslane. The highest and lowest relative growth rates were in larvae reared on Dill and Purslane, respectively. Furthermore, the highest amylolytic and proteolytic activities were in larvae fed with Coriander and Dill, respectively, while the lowest activities of these enzymes were on Purslane. In addition, correlation analysis revealed significant correlations between feeding characteristics and enzymatic activity of S. littoralis with biochemical compounds of the studied leafy vegetables. Our results suggest that Coriander is a suitable host, while Purslane displayed tolerance traits against S. littoralis, which can be used in sustainable management programs aiming to reduce chemical inputs.

Synergistic effects of chlorantraniliprole and camptothecin on physiological impairments, histopathological, biochemical changes, and genes responses in the larvae midgut of Spodoptera frugiperda.

Spodoptera frugiperda is an economically important agricultural pest and poses a serious threat to food security globally. Its management is gravely challenged by its high polyphagous nature, strong migratory ability, and massive fecundity. Chlorantraniliprole (CHL) is widely utilized in controlling S. frugiperda, its intensive application and over-reliance pose adverse health risks, development of resistance, toxicity to beneficial insects, natural enemies, and environmental contamination. To address S. frugiperda resistance to CHL and its inherent challenges, this study explores the synergistic effects of camptothecin (CPT) with CHL in its management. The binary mixed adversely induced the larvae weight and mortality when compared to single-treated. CHL + CPT (1:20 mg/L) had the highest larvae mortality of (73.80 %) with a high antagonistic factor (0.90), while (1:10 mg/L) with (66.10%) mortality exhibited a high synergistic factor (1.43). Further, CHL + CPT (1:10 mg/L) considerably altered the midgut epithelial cell, peritrophic membrane, microvilli, basement membrane, and regenerative cells. For biochemical analysis, CHL + CPT (1:10 mg/L) significantly decreased glutathione-S-transferase (1-chloro-2,4-dinitrobenzene CDNB) and cytochrome P450 (7-ethoxycoumarin O-deethylation) activities in the midgut in a dose and time dependent manner. Based on RNA-Seq analysis, a total of 4,373 differentially expressed genes (DEGs) were identified from the three treatments. CPT vs CK (Control) had 1694 (968 up-, 726 down-regulated), CHL vs CK with 1771 (978 up-, 793 down-regulated), and CHL + CPT vs CK had 908 (394 up-, 514 down-regulated) DEGs. The enrichment analysis disclosed significant pathways such as metabolism of xenobiotics by cytochrome P450, glutathione metabolism, TOLL and IMD (Immune Deficiency) signaling pathway, longevity regulating pathway. This study provides basis to expatiate on the molecular toxicological mechanism of CHL + CPT in management of fall armyworm.

First study on the root endophytic fungus Trichoderma hamatum as an entomopathogen: Development of a fungal bioinsecticide against cotton leafworm (Spodoptera littoralis).

Cotton leaf worm (Spodoptera littoralis) is a pest that produces important losses in horticultural and ornamental crops in greenhouse, being classified as quarantine pest A2 by EPPO. One of the strategies proposed to control agricultural pests in a health and environmentally friendly way is biological control with entomopathogenic fungi. The genus of filamentous fungi Trichoderma includes different species with direct (infection, antibiosis, anti-feeding, etc.) and indirect (systemic activation of plant defenses) insecticidal capacity, however, the species T. hamatum has never been described previously as entomopathogenic. In this work, the entomopathogenic capacity of T. hamatum on S. littoralis L3 larvae was analyzed by applying spores and fungal filtrates (topically and orally). Infection by spores was compared with the commercial entomopathogenic fungus Beauveria bassiana, obtaining similar results with respect to the production of larval mortality. Oral application of spores reported high mortality and fungal colonization of larvae, however, T. hamatum did not show chitinase activity when grown in the presence of S. littoralis tissues. Therefore, infection of S. littoralis larvae by T. hamatum is through natural openings such as mouth, anus or spiracles. With respect to the application of filtrates, only those obtained from the liquid culture of T. hamatum in contact with S. littoralis tissues reported a significant reduction in larval growth. Metabolomic analysis of the filtrates determined that the filtrate with insecticidal capacity presented the siderophore rhizoferrin in large quantities, which could be responsible for this activity. However, the production of this siderophore had never been previously described in Trichoderma and its insecticidal capacity was unknown. In conclusion, T. hamatum presents entomopathogenic capacity against S. littoralis larvae through the application of spores and filtrates, and both ways could be the basis for the development of efficient bioinsecticides against the pest.

NaMLP, a new identified Kunitz trypsin inhibitor regulated synergistically by JA and ethylene, confers Spodoptera litura resistance in Nicotiana attenuata.

KEY MESSAGE: We identified a miraculin-like protein (NaMLP) who is a new Kunitz trypsin inhibitor regulated synergistically by JA and ethylene signals and confers Spodoptera litura resistance in wild tobacco Nicotiana attenuata. The findings revealed a new source of trypsin inhibitor activities after herbivory, and provide new insights into the complexity of the regulation of trypsin inhibitor-based defense after insect herbivore attack. Upon insect herbivore attack, wild tobacco Nicotiana attenuata accumulates trypsin protease inhibitor (TPI) activities as a defense response from different protease inhibitor (PI) coding genes, including WRKY3-regulated NaKTI2, and JA-dependent NaPI. However, whether any other TPI gene exists in N. attenuata is still unclear. A miraculin-like protein gene (NaMLP) was highly up-regulated in N. attenuata after Alternaria alternata infection. However, silencing or overexpression of NaMLP had no effect on the lesion diameter developed on N. attenuata leaves after A. alternata inoculation. Meanwhile, the transcripts of NaMLP could be induced by wounding and amplified by Spodoptera litura oral secretions (OS). S. litura larvae gained significantly more biomass on NaMLP-silenced plants but less on NaMLP overexpressed plants. Although NaMLP showed low sequence similarity to NaKTI2, it had conserved reaction sites of Kunitz trypsin inhibitors, and exhibited TPI activities when its coding gene was overexpressed transiently or stably in N. attenuata. This was consistent with the worst performance of S. litura larvae on NaMLP overexpressed lines. Furthermore, NaMLP-silenced plants had reduced TPI activities and better S. litura performance. Finally, OS-elicited NaMLP was dramatically reduced in JA-deficient AOC silencing and ethylene-reduced ACO-silencing plants, and the expression of NaMLP could be significantly induced by methyl jasmonate or ethephon alone, but dramatically amplified by co-treatment of both methyl jasmonate and ethephon. Thus, our results demonstrate that in addition to JA-regulated NaPI, and WRKY3/6-dependent NaKTI2, N. attenuata plants also up-regulates TPI activities via NaMLP, which confers S. litura resistance through JA and ethylene signaling pathways in a synergistic way.

The green lacewing Chrysopa formosa as a potential biocontrol agent for managing Spodoptera frugiperda and Spodoptera litura.

Understanding predator-prey interactions is essential for successful pest management by using predators, especially for the suppression of novel invasive pest. The green lacewing Chrysopa formosa is a promising polyphagous predator that is widely used in the biocontrol of various pests in China, but information on the control efficiency of this predator against the seriously invasive pest Spodoptera frugiperda and native Spodoptera litura is limited. Here we evaluated the predation efficiency of C. formosa adults on eggs and first- to third-instar larvae of S. frugiperda and S. litura through functional response experiments and determined the consumption capacity and prey preference of this chrysopid. Adults of C. formosa had a high consumption of eggs and earlier instar larvae of both prey species, and displayed a type II functional response on all prey stages. Attack rates of the chrysopid on different prey stages were statistically similar, but the handling time increased notably as the prey developed. The highest predation efficiency and shortest-handling time were observed for C. formosa feeding on Spodoptera eggs, followed by the first-instar larvae. C. formosa exhibited a significant preference for S. litura over S. frugiperda in a two-prey system. In addition, we summarized the functional response and predation efficiency of several chrysopids against noctuid pests and made a comparison with the results obtained from C. formosa. These results indicate that C. formosa has potential as an agent for biological control of noctuid pests, particularly for the newly invasive pest S. frugiperda in China.

Responses of the fall armyworm (Spodoptera frugiperda) to different host plants: Implications for its management strategy.

BACKGROUND: The selection of suitable host plants for oviposition is critical for herbivorous insects to maximise survival of their offspring. Olfaction plays an important role in this process. However, little is known about how olfaction shapes the interaction between the fall armyworm (Spodoptera frugiperda) and host plants. In this study, we tested the hypothesis that olfaction guides the host selection process in the fall armyworm using oviposition and wind tunnel bioassays. RESULTS: In no-choice and dual-choice assays, female moths oviposited on all seven host plants that were tested (maize, sorghum, wheat, bean, cowpea, tomato and cabbage). However, in multiple-choice assays, no eggs were deposited on cowpea and cabbage. We found that maize, sorghum and wheat were most preferred for oviposition, whereas cowpea was least preferred. Wind tunnel assays confirmed these divergent oviposition preferences, with maize, sorghum and wheat odours being the most attractive. Gas chromatography-mass spectrometry analysis followed by random forest classification identified terpenes as the potential host-plant attractants. CONCLUSION: Our results improve our understanding of the chemical ecology of the fall armyworm and suggest that some of these host plants could offer potential for use in an intercropping strategy to manage S. frugiperda. © 2022 Society of Chemical Industry.