Dominant strain shift in the invasive fall armyworm (Lepidoptera: Noctuidae) populations in Thailand as inferred from mitochondrial COI and nuclear Tpi genes.

The fall armyworm, Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae), is a significant global pest, that exhibits 2 discernible strains, corn strain (CS) and rice strain (RS). After initial detection in the eastern hemisphere in 2016, the dominant strain was identified as RS based only on cytochrome C oxidase subunit I (COI) mitochondrial gene from limited samples from various countries, including Thailand. This study aimed to assess strain and haplotype variation in the S. frugiperda populations in Thailand using both mitochondrial COI and nuclear triosephosphate isomerase (Tpi) genes. Analyses of COI sequences (n = 105) revealed 2 predominant haplotypes, COICSh4 (82.86%) and COIRSh1 (17.14%), and the analyses of Tpi sequences (n = 99) revealed 6 haplotypes, with TpiCa1a (53.53%) being the most prevalent. Of the 98 caterpillar samples, the majority exhibited true CS (83.67%) for both genes. Meanwhile, interstrain hybrids, indicated by gene discordance, accounted for the minority (16.33%). Interestingly, despite the initial dominance of RS during the 2018 outbreak, the current study identified CS as the prevalent strain across all localities in Thailand. These findings suggested a shift in S. frugiperda dynamics in Thailand that was possibly influenced by factors, such as competitive exclusion principle, pesticide usage in rice cultivation, and preferences for corn over rice. Our study suggests a need to reexamine the previous reports of rice-strain dominance in various countries in the eastern hemisphere after the initial invasion.

Estrogen-related receptor, a molecular target against lepidoptera pests.

Until recently, chemical pesticides were one of the most effective means of controlling agricultural pests; therefore, the search for insecticide targets for agricultural pests has been an ongoing problem. Estrogen-related receptors (ERRs) are transcription factors that regulate cellular metabolism and energy homeostasis in animals. Silkworms are highly sensitive to chemical pesticides, making them ideal models for pesticide screening and evaluation. In this study, we detected ERR expression in key organs involved in pesticide metabolism in silkworms (Bombyx mori), including the fat body and midgut. Using ChIP-seq technology, many estrogen- related response elements were identified in the 2000-bp promoter region upstream of metabolism-related genes, almost all of which were potential ERR target genes. The ERR inhibitor, XCT-790, and the endocrine disruptor, bisphenol A, significantly inhibited expression of the ERR target genes, BmTreh-1, BmTret-1, BmPK, BmPFK, and BmHK, in the fat bodies of silkworms, resulting in pupation difficulties in silkworm larvae that ultimately lead to death. In addition, based on the clarification that the ERR can bind to XCT-790, as observed through biofilm interferometry, its three-dimensional spatial structure was predicted, and using molecular docking techniques, small-molecule compounds with a stronger affinity for the ERR were identified. In summary, utilizing the powerful metabolic regulatory function of ERR in Lepidoptera pests, the developed small molecule inhibitors of ERR can be used for future control of Lepidoptera pests.

Biochemical characterization of Bombyx mori Dicer-2 that dices double-stranded RNAs into 20-nt small RNA.

We detected enzymatic activity that generates 20-nucleotide (nt) RNA from double-stranded RNAs (dsRNAs) in crude extracts prepared from various silkworm (Bombyx mori) organs. The result using knocked-down cultured cells indicated that this dicing activity originated from B. mori Dicer-2 (BmDcr2). Biochemical analyses revealed that BmDcr2 preferentially cleaves 5'-phosphorylated dsRNAs at the 20-nt site-counted from the 5'-phosphorylated end-and required ATP and magnesium ions for the dicing reaction. This is the first report of the biochemical characterization of Dicer-2 in lepidopteran insects. This enzymatic property of BmDcr2 in vitro is consistent with the in vivo small interfering RNA profile in virus-infected silkworm cells.

Historical Note on the Genus Trichogramma (Hymenoptera, Trichogrammatidae) in Brazil, Focusing on Taxonomy and Diversity.

The history of the taxonomy of Trichogramma parasitoid wasps can be divided into two phases. In the first phase, species identifications were based on external morphological characters; however, these identifications proved to be confusing and inaccurate. In the second phase, starting in the 1970s, taxa were identified based on charcteristics of the male genital capsule , leading to a major advance in Trichogramma taxonomy. The history of Trichogramma taxonomy in Brazil is recent and mainly related to species that parasitize agricultural pests. In Brazil, the first phase of Trichogramma taxonomy occurred in the 1960s, while the second phase occurred from the 1980s onward. In this second phase, Trichogramma taxonomy progressed significantly and knowledge of Trichogramma diversity as well as associations with lepidopteran pests improved markedly in Brazil as well as worldwide. The last five decades have seen significant progress in studies in Brazil, with taxonomy evolving from identifications based exclusively on morphological characters to integrative taxonomy, encompassing biology (crosses) and morphometry. This historical outline presents the phases of Trichogramma taxonomy in Brazil, addressing the hurdles encountered in the first descriptions, erroneous records of the species, and species descriptions since the 1980s. We highlight the importance of accurately identifying Trichogramma taxa for their use in biological control, as well as species diversity and associations with lepidopteran hosts.

Phytometabolites from coral jasmine flower extracts: Toxic effects on Spodoptera litura and enzyme inhibition in nontarget earthworm Eisenia fetida as an alternative approach.

Green pesticides, derived from natural sources, have gained wider attention as an alternative to synthetic pesticides for managing polyphagous pests, such as Spodoptera litura. In this study, the methanolic flower extract of Nyctanthes arbor-tristis (Mx-Na-t) was subjected to chemical screening, and 3-hydroxy-1,2-dimethyl-4(1H)-pyridone (3H-dp) and tyrosol (Ty-ol) were identified as the major derivatives. The toxic effects of Mx-Na-t (500 ppm) were highest in third-instar S. litura larvae (96.4%), while those of 3H-dp and Ty-ol (5 ppm) were highest in second-instar larvae (76.5% and 81.4%, respectively). The growth and development of S. litura larvae and pupae were significantly reduced by all three treatments. Fecundity rates were also reduced by all treatments [from 1020 eggs (control) to 540 eggs by Mx-Na-t treatment, 741 eggs by 3H-dp treatment, and 721 eggs by Ty-ol treatment]. The extract and its active constituents decreased adult emergence and slowed total larval development in a dose-dependent manner. A decrease was noted in the major gut enzymes of young S. litura larvae exposed to Mx-Na-t, 3H-dp, and Ty-ol. Moreover, midgut tissues of fourth-instar larvae were severely damaged by Mx-Na-t (250 ppm), 3H-dp (2.5 ppm), and Ty-ol (2.5 ppm); the treatments induced structural damage to the epithelial cells and gut lumen. The earthworm Eisenia fetida was used to assess nontarget toxicity. Compared with cypermethrin, the phytochemicals exhibited minimal effects on the earthworm's detoxifying enzymes superoxide dismutase and catalase after 14 days of treatment. Moreover, in silico predictions using BeeTox and ProTox-II indicated little or no toxicity of 3H-dp and Ty-ol toward honey bees and other nontarget species.

Structural Insights into the Substrate Binding of Farnesyl Diphosphate Synthase FPPS1 from Silkworm, Bombyx mori.

Farnesyl diphosphate synthase (FPPS) is an important enzyme involved in the juvenile hormone (JH) biosynthesis pathway. Herein, we report the crystal structure of a type-I Lepidopteran FPPS from Bombyx mori (BmFPPS1) at 2.80 Å resolution. BmFPPS1 adopts an α-helix structure with a deep cavity at the center of the overall structure. Computational simulations combined with biochemical analysis allowed us to define the binding mode of BmFPPS1 to its substrates. Structural comparison revealed that BmFPPS1 adopts a structural pattern similar to that of type-II FPPS but exhibits a distinct substrate-binding site. These findings provide a structural basis for understanding substrate preferences and designing FPPS inhibitors. Furthermore, the expression profiles and RNA interference of BmFPPSs indicated that they play critical roles in the JH biosynthesis and larval-pupal metamorphosis. These findings enhance our understanding of the structural features of type-I Lepidopteran FPPS while providing direct evidence for the physiological role of BmFPPSs in silkworm development.

Expression of the Wolbachia male-killing factor Oscar impairs dosage compensation in lepidopteran embryos.

Wolbachia are intracellular bacteria in insects that can manipulate the sexual development and reproduction by male killing or other methods. We have recently identified a Wolbachia protein named Oscar that acts as a male-killing factor for lepidopteran insects. Oscar interacts with the Masculinizer (Masc) protein, which is required for both masculinization and dosage compensation (DC) in lepidopteran insects. Embryonic expression of Oscar inhibits masculinization and causes male killing in two lepidopteran species, Ostrinia furnacalis and Bombyx mori. However, it remains unknown whether Oscar-induced male killing is caused by a failure of DC. Here, we performed a transcriptome analysis of Oscar complementary RNA-injected O. furnacalis and B. mori embryos, and found that Oscar primarily targets the Masc protein, resulting in male killing by interfering with DC in lepidopteran insects.

Development of a soybean leaf disc assay for determining oral insecticidal activity in the lepidopteran agricultural pest Helicoverpa armigera.

Pest insects pose a heavy burden on global agricultural industries with small molecule insecticides being predominantly used for their control. Unwanted side effects and resistance development plagues most small molecule insecticides such as the neonicotinoids, which have been reported to be harmful to honeybees. Bioinsecticides like Bacillus thuringiensis (Bt) toxins can be used as environmentally-friendly alternatives. Arachnid venoms comprise another promising source of bioinsecticides, containing a multitude of selective and potent insecticidal toxins. Unfortunately, no standardised insect models are currently available to assess the suitability of insecticidal agents under laboratory conditions. Thus, we aimed to develop a laboratory model that closely mimics field conditions by employing a leaf disk assay (LDA) for oral application of insecticidal agents in a bioassay tray format. Neonate larvae of the cotton bollworm (Helicoverpa armigera) were fed with soybean (Glycine max) leaves that were treated with different insecticidal agents. We observed dose-dependent insecticidal effects for Bt toxin and the neonicotinoid insecticide imidacloprid, with imidacloprid exhibiting a faster response. Furthermore, we identified several insecticidal arachnid venoms that were active when co-applied with sub-lethal doses of Bt toxin. We propose the H. armigera LDA as a suitable tool for assessing the insecticidal effects of insecticidal agents against lepidopterans.

Cas9-Mediated Gene Editing Using Receptor-Mediated Ovary Transduction of Cargo (ReMOT) Control in Bombyx mori.

Lepidoptera is one of the most speciose insect orders, causing enormous damage to agricultural and forest crops. Although genome editing has been achieved in a few Lepidoptera for insect controls, most techniques are still limited. Here, by injecting female pupae of the Lepidoptera model species, Bombyx mori, gene editing was established using the Receptor-Mediated Ovary Transduction of Cargo (ReMOT) control technique. We identified a B. mori oocytes-targeting peptide ligand (BmOTP, a 29 aa of vitellogenin N-terminal of silkworms) with a highly conserved sequence in lepidopteran insects that could efficiently deliver mCherry into oocytes. When BmOTP was fused to CRISPR-associated protein 9 (Cas9) and the BmOTP-Cas9 ribonucleoprotein complex was injected into female pupae, heritable editing of the offspring was achieved in the silkworms. Compared with embryo microinjection, individual injection is more convenient and eliminates the challenge of injecting extremely small embryos. Our results will significantly facilitate the genetic manipulation of other lepidopteran insects, which is essential for advancing lepidopteran pest control.

Mechanisms and roles of the first stage of nodule formation in lepidopteran insects.

Nodule formation is a process of cellular immunity in insects and other arthropods with open circulatory systems. Based on histological observations, nodule formation occurs in 2 stages. The first stage occurs immediately after microbial inoculation and includes aggregate formation by granulocytes. The second stage occurs approximately 2-6 h later and involves the attachment of plasmatocytes to melanized aggregates produced during the first stage. The first stage response is thought to play a major role in the rapid capture of invading microorganisms. However, little is known regarding how granulocytes in the hemolymph form aggregates, or how the first stage of the immunological response protects against invading microorganisms. Since the late 1990s, our understanding of the molecules and immune pathways that contribute to nodule formation has improved. The first stage of nodule formation involves a hemocyte-induced response that is triggered by pathogen-associated molecular pattern (PAMP) recognition proteins in the hemolymph regulated by a serine proteinase cascade and cytokine (Spätzle) and Toll signaling pathways. Hemocyte agglutination proceeds through stepwise release of biogenic amine, 5-HT, and eicosanoids that act downstream of the Toll pathway. The first stage of nodule formation is closely linked to melanization and antimicrobial peptide (AMP) production, which is critical for insect humoral immunity. Nodule formation in response to artificial inoculation with millions of microorganisms has long been studied. It has recently been suggested that this system is the original natural immune system, and enables insects to respond to a single invading microorganism in the hemocoel.

More than 10 years after commercialization, Vip3A-expressing MIR162 remains highly efficacious in controlling major Lepidopteran maize pests: laboratory resistance selection versus field reality.

MIR162, a maize event that expresses Vip3Aa20 (Vip3A) approved for commercial cultivation around 2010, has been excellent for control of major Lepidopteran pests. However, development of fall armyworm (FAW) resistance to Vip3A is a serious concern. Resistant colonies selected in the laboratory can serve as valuable tools not only for better understanding of Vip3A's mode of action (MOA) and mechanism of resistance (MOR) but also for screening novel leads of new MOA that will help control FAW in case resistance to Vip3A in the field becomes a reality. We selected a Vip3A-resistant FAW strain, FAWVip3AR, by subjecting a FAW founder population containing field genetics to Vip3A exposure. FAWVip3AR had >9800-fold resistance to Vip3A by diet surface overlay bioassays and resistance was stable. Feeding bioassays using detached leaf tissues or whole plants indicated that FAWVip3AR larvae readily fed and completed the full life cycle on Vip3A-expressing MIR162 maize plants and leaf tissues that killed 100% of susceptible larvae. Yet, FAWVip3AR faced at least two challenges. First, FAWVip3AR suffered an apparent disadvantage (incomplete resistance) when feeding on MIR162 in comparison to FAWVip3AR feeding on Vip3A-free isoline AX5707 maize; and second, FAWVip3AR showed a fitness costs in comparison to a Vip3A-susceptible strain when both fed on AX5707. We also demonstrated that, >10 years after commercialization, MIR162 and Vip3A remain highly efficacious against field populations of three major Lepidopteran pests from different geographic locations and FAW strains resistant to other Bacillus thuringiensis (Bt) toxins that are currently on the market.

Genomic-proteomic analysis of a novel Bacillus thuringiensis strain: toxicity against two lepidopteran pests, abundance of Cry1Ac5 toxin, and presence of InhA1 virulence factor.

Bacillus thuringiensis (Bt) is a biological alternative to the indiscriminate use of chemical insecticides in agriculture. Due to resistance development on insect pests to Bt crops, isolating novel Bt strains is a strategy for screening new pesticidal proteins or strains containing toxin profile variety that can delay resistance. Besides, the combined genomic and proteomic approaches allow identifying pesticidal proteins and virulence factors accurately. Here, the genome of a novel Bt strain (Bt TOL651) was sequenced, and the proteins from the spore-crystal mixture were identified by proteomic analysis. Toxicity bioassays with the spore-crystal mixture against larvae of Diatraea saccharalis and Anticarsia gemmatalis, key pests of sugarcane and soybean, respectively, were performed. The toxicity of Bt TOL651 varies with the insect; A. gemmatalis (LC50 = 1.45 ng cm-2) is more susceptible than D. saccharalis (LC50 = 73.77 ng cm-2). Phylogenetic analysis of the gyrB gene indicates that TOL651 is related to Bt kenyae strains. The genomic analysis revealed the presence of cry1Aa18, cry1Ac5, cry1Ia44, and cry2Aa9 pesticidal genes. Virulence factor genes such as phospholipases (plcA, piplc), metalloproteases (inhA), hemolysins (cytK, hlyIII, hblA, hblC, hblD), and enterotoxins (nheA, nheB, nheC) were also identified. The combined use of the genomic and proteomic data indicated the expression of Cry1Aa18, Cry1Ac5, and Cry2Aa9 proteins, with Cry1Ac5 being the most abundant. InhA1 also was expressed and may contribute to Bt TOL651 pathogenicity. These results provide Bt TOL651 as a new tool for the biocontrol of lepidopteran pests.

Tree dissimilarity determines multi-dimensional beta-diversity of herbivores and carnivores via bottom-up effects.

Global biodiversity decline and its cascading effects through trophic interactions pose a severe threat to human society. Establishing the impacts of biodiversity decline requires a more thorough understanding of multi-trophic interactions and, more specifically, the effects that loss of diversity in primary producers has on multi-trophic community assembly. Within a synthetic conceptual framework for multi-trophic beta-diversity, we tested a series of hypotheses on neutral and niche-based bottom-up processes in assembling herbivore and carnivore communities in a subtropical forest using linear models, hieratical variance partitioning based on linear mixed-effects models (LMMs) and simulation. We found that the observed taxonomic, phylogenetic and functional beta-diversity of both herbivorous caterpillars and carnivorous spiders were significantly and positively related to tree dissimilarity. Linear models and variance partitioning for LMMs jointly suggested that as a result of bottom-up effects, producer dissimilarities were predominant in structuring consumer dissimilarity, the strength of which highly depended on the trophic dependencies on producers, the diversity facet examined, and data quality. Importantly, linear models for standardized beta-diversities against producer dissimilarities implied a transition between niche-based processes such as environmental filtering and competitive exclusion, which supports the role of bottom-up effect in determining consumer community assembly. These findings enrich our mechanistic understanding of the 'Diversity Begets Diversity' hypothesis and the complexity of higher-trophic community assembly, which is fundamental for sustainable biodiversity conservation and ecosystem management.

A female-biased odorant receptor tuned to the lepidopteran sex pheromone in parasitoid Microplitis mediator guiding habitat of host insects.

INTRODUCTION: The parasitoid wasp Microplitis mediator is an important natural enemy of the turnip moth Agrotis segetum and other Noctuidae pests. In our field observation, it was fortuitously discovered that sex pheromone traps used for A. segetum also attract female wasps, verified by a simulated field condition dual-choice laboratory assay. Therefore, it was hypothesized that olfactory recognition could be crucial in this process. In this regard, a female-biased odorant receptor of the wasp, MmedOR49, attracted our attention. OBJECTIVES: To unravel the significance of the female-biased MmedOR49 regulating host pheromone recognition. METHODS: Expression analysis (fluorescence in situ hybridization; quantitative realtime PCR), in vitro (two-electrode voltage-clamp recordings) and in vivo (RNAi combined with behavioral assessments) functional studies, and bioinformatics (structural modeling and molecular docking) were carried out to investigate the characteristics of MmedOR49. RESULTS: MmedOR49 expression was detected in the antennae of females by FISH. Quantification indicated that the expression level of MmedOR49 increased significantly after adult emergence. In vitro functional study revealed that MmedOR49 was specifically tuned to cis-5-decenyl acetate (Z5-10:Ac), the major sex pheromone component of A. segetum. Molecular docking showed that Z5-10:Ac strongly bound to the key amino acid residues His 80, Ile 81, and Arg 84 of MmedOR49 through hydrogen bonding. Behavioral assays indicated that female wasps were significantly attracted by Z5-10:Ac in a three-cage olfactometer. RNAi targeting further confirmed that MmedOR49 was necessary to recognize Z5-10:Ac, as female wasps lost their original behavioral responses to Z5-10:Ac after down-regulation of the MmedOR49 transcript. CONCLUSION: Although M. mediator is a larval endoparasitoid, female wasps have a behavioral preference for a sex pheromone component of lepidopteran hosts. In this behavior, for female M. mediator, MmedOR49 plays an important role in guiding the habitat of host insects. These data provide a potential target for enhancing natural enemy utilization and pest control.

Bt maize can provide non-chemical pest control and enhance food safety in China.

China is the world's second-largest maize producer and consumer. In recent years, the invasive fall armyworm Spodoptera frugiperda (J.E. Smith) has adversely affected maize productivity and compromised food security. To mitigate pest-inflicted food shortages, China's Government issued biosafety certificates for two genetically modified (GM) Bt maize hybrids, Bt-Cry1Ab DBN9936 and Bt-Cry1Ab/Cry2Aj Ruifeng 125, in 2019. Here, we quantitatively assess the impact of both Bt maize hybrids on pest feeding damage, crop yield and food safety throughout China's maize belt. Without a need to resort to synthetic insecticides, Bt maize could mitigate lepidopteran pest pressure by 61.9-97.3%, avoid yield loss by 16.4-21.3% (range -11.9-99.2%) and lower mycotoxin contamination by 85.5-95.5% as compared to the prevailing non-Bt hybrids. Yield loss avoidance varied considerably between experimental sites and years, as mediated by on-site infestation pressure and pest identity. For either seed mixtures or block refuge arrangements, pest pressure was kept below established thresholds at 90% Bt maize coverage in Yunnan (where S. frugiperda was the dominant species) and 70% Bt maize coverage in other sites dominated by Helicoverpa armigera (Hübner) and Ostrinia furnacalis (Guenée). Drawing on experiences from other crop/pest systems, Bt maize in se can provide area-wide pest management and thus, contribute to a progressive phase-down of chemical pesticide use. Hence, when consciously paired with agroecological and biodiversity-based measures, GM insecticidal crops can ensure food and nutrition security, contribute to the sustainable intensification of China's agriculture and reduce food systems' environmental footprint.

Ataxia Telangiectasia-Mutated Is Activated but Not Required for Productive Autographa californica Multiple Nucleopolyhedrovirus Infection.

Multiplication of the invertebrate DNA baculoviruses activates the host DNA damage response (DDR), which promotes virus DNA replication. DDR signaling is initiated by the host insect's phosphatidylinositol-3 kinase-related kinases (PIKKs), including ataxia telangiectasia-mutated kinase (ATM). Like other PIKKs, ATM phosphorylates an array of host DDR proteins at serine/threonine glutamine (S/TQ) motifs, the result of which leads to cell cycle arrest, DNA repair, or apoptosis. To define the role of host PIKKs in baculovirus replication, we compared replication levels of the baculovirus prototype species Autographa californica multiple nucleopolyhedrovirus in permissive Spodoptera frugiperda (SF21) cells with and without ATM function. Caffeine, which inhibits multiple DDR kinases, and the ATM-specific inhibitors KU-55933 and KU-60019 each prevented phosphorylation of Spodoptera histone H2AX (SfH2AX), a recognized indicator of ATM activity. However, only caffeine reduced autographa californica multiple nucleopolyhedrovirus (AcMNPV)-induced bulk phosphorylation of S/TQ protein motifs. Furthermore, only caffeine, not KU-55933 or KU-60019, reduced AcMNPV yields, suggesting a limited role for ATM. To investigate further, we identified and edited the Spodoptera ATM gene (sfatm). Consistent with ATM's known functions, CRISPR/Cas9-mediated knockout of sfatm eliminated DNA damage-induced phosphorylation of DDR marker SfH2AX in SF21 cells. However, loss of sfatm failed to affect the levels of AcMNPV multiplication. These findings suggested that in the absence of the kinase SfATM, another caffeine-sensitive host DDR kinase promotes S/TQ phosphorylation and baculovirus multiplication. Thus, baculoviruses activate and utilize the host insect DDR in an ATM-independent manner. IMPORTANCE The DDR, while necessary for the maintenance and fidelity of the host genome, represents an important cellular response to viral infection. The prolific DNA baculoviruses activate and manipulate the invertebrate DDR by using mechanisms that positively impact virus multiplication, including virus DNA replication. As the key DDR initiator kinase, ATM was suspected to play a critical role in this host response. However, we show here that baculovirus AcMNPV activates an ATM-independent DDR. By identifying the insect host ATM ortholog (Spodoptera frugiperda SfATM) and evaluating genetic knockouts, we show that SfATM is dispensable for AcMNPV activation of the DDR and for virus replication. Thus, another PIKK, possibly the closely related kinase ATR (ATM- and Rad3-related kinase), is responsible for efficient baculovirus multiplication. These findings better define the host pathways used by invertebrates to engage viral pathogens, including DNA viruses.

Effects of Nutritional Composition of Different Prey Eggs on Development and Reproduction of the Predatory Bug, Orius sauteri (Hemiptera: Anthocoridae).

Orius sauteri (Poppius) is an important predator of many economically important insect pests. The mass rearing of O. sauteri is difficult, limiting its application in pest control. Here we assessed the nutritional quality of eggs of Sitotroga cerealella (Olivier), Agrotis ypsilon (Rottemberg), or Spodoptera litura (Fabricius), and their potential for rearing O. sauteri in the laboratory for two generations. Of species tested, S. cerealella eggs resulted in the highest survival and reproduction of O. sauteri compared to the other two lepidopteran species. Eggs of A. ypsilon were a suitable diet for the nymphal stage, which developed faster on A. ysilon eggs than those of S. cerealella eggs. Conversely, eggs of S. litura were not a suitable diet for O. sauteri, and they disrupted the development and reproduction of O. sauteri. Sitotroga cerealella eggs showed advantages in all the nutritional components evaluated. Orius sauteri fed S. litura eggs contained significantly lower protein levels than those fed on the other eggs tested. Spodoptera litura eggs significantly enhanced the CAT activity in O. sauteri, which suggests that some components from S. litura eggs harmed the development and reproduction of O. sauteri. Based on these results, we suggest using a combined diet for mass rearing of the pirate bug, feeding the nymphs and adults with A. ypsilon eggs and S. cerealella eggs, respectively. This study contributes to the discovery of artificial diets for mass rearing O. sauteri and other Orius species in the future.

Mutation of a lepidopteran-specific PMP-like protein, BmLSPMP-like, induces a stick body shape in silkworm, Bombyx mori.

BACKGROUND: Lepidoptera is one of the largest orders of insects, some of which are major pests of crops and forests. The cuticles of lepidopteran pests play important roles in defense against insecticides and pathogens, and are indispensable for constructing and maintaining extracellular structures and locomotion during their life cycle. Lepidopteran-specific cuticular proteins could be potential targets for lepidopteran pest control. But information on this is limited. Our research aimed to screen the lepidopteran-specific cuticular proteins using the lepidopteran model, the silkworm, to explore the molecular mechanism underlying the involvement of cuticular proteins in body shape construction. RESULTS: Positional cloning showed that BmLSPMP-like, a gene encoding a lepidopteran-specific peritrophic matrix protein (PMP) like protein which includes a peritrophin A-type chitin-binding domain (CBM_14), is responsible for the stick (sk) mutation. BmLSPMP-like is an evolutionarily conserved gene that exhibits synteny in Lepidoptera and underwent purifying selection during evolution. Expression profiles demonstrated that BmLSPMP-like is expressed in chitin-forming tissues, testis and ovary, and accumulates in the cuticle. BmLSPMP-like knockout, generated with CRISPR/Cas9, resulted in a stick-like larval body shape phenotype. Over-expression of BmLSPMP-like in the sk mutant rescued its abnormal body shape. The results showed that BmLSPMP-like may be involved in assemblage in the larval cuticle. CONCLUSION: Our results suggested that the dysfunction of BmLSPMP-like may result in a stick body shape phenotype in silkworm, through the regulation of the arrangement of the chitinous laminae and cuticle thickness. Our study provides new evidence of the effects of LSPMP-likes on lepidopteran body shape formation, metamorphosis and mortality, which could be an eco-friendly target for lepidopteran pest management. © 2022 Society of Chemical Industry.

Chromosomal-level reference genome of the moth Heortia vitessoides (Lepidoptera: Crambidae), a major pest of agarwood-producing trees.

The moth Heortia vitessoides Moore (Lepidoptera: Crambidae) is a major pest of ecologically, commercially and culturally important agarwood-producing trees in the genus Aquilaria. In particular, H. vitessoides is one of the most destructive defoliating pests of the incense tree Aquilaria sinesis, which produces a valuable fragrant wood used as incense and in traditional Chinese medicine [33]. Nevertheless, a genomic resource for H. vitessoides is lacking. Here, we present a chromosomal-level assembly for H. vitessoides, consisting of a 517 megabase (Mb) genome assembly with high physical contiguity (scaffold N50 of 18.2 Mb) and high completeness (97.9% complete BUSCO score). To aid gene annotation, 8 messenger RNA transcriptomes from different developmental stages were generated, and a total of 16,421 gene models were predicted. Expansion of gene families involved in xenobiotic metabolism and development were detected, including duplications of cytosolic sulfotransferase (SULT) genes shared among lepidopterans. In addition, small RNA sequencing of 5 developmental stages of H. vitessoides facilitated the identification of 85 lepidopteran conserved microRNAs, 94 lineage-specific microRNAs, as well as several microRNA clusters. A large proportion of the H. vitessoides genome consists of repeats, with a 29.12% total genomic contribution from transposable elements, of which long interspersed nuclear elements (LINEs) are the dominant component (17.41%). A sharp decrease in the genome-wide percentage of LINEs with lower levels of genetic distance to family consensus sequences suggests that LINE activity has peaked in H. vitessoides. In contrast, opposing patterns suggest a substantial recent increase in DNA and LTR element activity. Together with annotations of essential sesquiterpenoid hormonal pathways, neuropeptides, microRNAs and transposable elements, the high-quality genomic and transcriptomic resources we provide for the economically important moth H. vitessoides provide a platform for the development of genomic approaches to pest management, and contribute to addressing fundamental research questions in Lepidoptera.

Lethal and sublethal effects of chlorantraniliprole on the migratory moths Agrotis ipsilon and A. segetum: New perspectives for pest management strategies.

BACKGROUND: Agrotis ipsilon and A. segetum are major migratory pests of many crops in China, and frequent regional outbreaks cause severe yield losses. Use of food attractants is one of the most promising control methods against adult lepidoptera, notably through the attract-and-kill strategy. Chlorantraniliprole's acute toxicity and sublethal effects on both moths were evaluated. RESULTS: Chlorantraniliprole showed high activity against both adults of both species, with LC20 and LC50 values of 0.08 and 0.21 mg L-1 (A. ipsilon), and 0.14 and 0.51 mg L-1 (A. segetum). The fecundity, effective oviposition rate, and egg hatching rate of both species in dual-sex exposure treatments were all significantly reduced compared with the control, and the population growth coefficients in the LC50 ♀ × LC50 ♂ treatments were only 0.32% (A. ipsilon) and 3.35% (A. segetum) that of the control. Furthermore, the flight distance was significantly suppressed from 6.67 km (control) to 0.01 km (LC50 ) for A. ipsilon, and from 7.39 km (control) to 0.78 km (LC50 ) for A. segetum. The proportions of robust- and medium-flight individuals of A. ipsilon and A. segetum in exposure treatments were greatly reduced. CONCLUSIONS: Low lethal concentration exposures to chlorantraniliprole can drastically reduce the reproduction and flight performance of A. ipsilon and A. segetum, while inhibiting the production of offspring, suggesting chlorantraniliprole would be an excellent compound for use in combination with food attractants. Chlorantraniliprole has good potential for management of the two long-range migratory pests tested using an attract-and-kill strategy. © 2022 Society of Chemical Industry.