Nitrogen-mediated volatilisation of defensive metabolites in tomato confers resistance to herbivores.

Plants synthesise a vast array of volatile organic compounds (VOCs), which serve as chemical defence and communication agents in their interactions with insect herbivores. Although nitrogen (N) is a critical resource in the production of plant metabolites, its regulatory effects on defensive VOCs remain largely unknown. Here, we investigated the effect of N content in tomato (Solanum lycopersicum) on the tobacco cutworm (Spodoptera litura), a notorious agricultural pest, using biochemical and molecular experiments in combination with insect behavioural and performance analyses. We observed that on tomato leaves with different N contents, S. litura showed distinct feeding preference and growth and developmental performance. Particularly, metabolomics profiling revealed that limited N availability conferred resistance upon tomato plants to S. litura is likely associated with the biosynthesis and emission of the volatile metabolite α-humulene as a repellent. Moreover, exogenous application of α-humulene on tomato leaves elicited a significant repellent response against herbivores. Thus, our findings unravel the key factors involved in N-mediated plant defence against insect herbivores and pave the way for innovation of N management to improve the plant defence responses to facilitate pest control strategies within agroecosystems.

Tetraniliprole risk assessment: Unveiling a hidden threat for managing a generalist herbivore.

Insecticide resistance poses a significant challenge in managing generalist herbivores such as the tobacco cutworm (TCW), Spodoptera litura. This study investigates the potential risks associated with using the novel diamide insecticide tetraniliprole to control TCW. A tetraniliprole-resistant strain was developed through twelve generations of laboratory selection, indicating an intermediate risk of resistance development. Field monitoring in China revealed a significant incidence of resistance, particularly in the Nanchang (NC) population (>100-fold). Tetraniliprole showed moderate to high cross-resistance to multiple insecticides and was autosomally inherited with incomplete dominance, controlled by multiple genes, some of which belong to the cytochrome P450 family associated with enhanced detoxification. Life table studies indicated transgenerational hormesis, stimulating TCW female fecundity and increasing population net reproduction rates (R0). These findings suggest a potential for pest resurgence under tetraniliprole use. The integrated risk assessment provides a basis for the sustainable management of TCW using tetraniliprole.

Chromosome-level genome of black cutworm provides novel insights into polyphagy and seasonal migration in insects.

BACKGROUND: The black cutworm, Agrotis ipsilon, is a serious global underground pest. Its distinct phenotypic traits, especially its polyphagy and ability to migrate long distances, contribute to its widening distribution and increasing difficulty of control. However, knowledge about these traits is still limited. RESULTS: We generated a high-quality chromosome-level assembly of A. ipsilon using PacBio and Hi-C technology with a contig N50 length of ~ 6.7 Mb. Comparative genomic and transcriptomic analyses showed that detoxification-associated gene families were highly expanded and induced after insects fed on specific host plants. Knockout of genes that encoded two induced ABC transporters using CRISPR/Cas9 significantly reduced larval growth rate, consistent with their contribution to host adaptation. A comparative transcriptomic analysis between tethered-flight moths and migrating moths showed expression changes in the circadian rhythm gene AiCry2 involved in sensing photoperiod variations and may receipt magnetic fields accompanied by MagR and in genes that regulate the juvenile hormone pathway and energy metabolism, all involved in migration processes. CONCLUSIONS: This study provides valuable genomic resources for elucidating the mechanisms involved in moth migration and developing innovative control strategies.

Potential host range of Cotesia vanessae (Hymenoptera: Braconidae), a parasitoid new to North America and a possible biological control agent of noctuid pest species.

The likelihood of parasitoids establishing in new geographic regions depends upon the availability of suitable host species. Identifying these hosts and the degree of their suitability is particularly important when they include species that are economically important as pests. In laboratory studies, we examined the suitability of 47 species of Lepidoptera as potential hosts of a parthenogenetic strain of the gregarious parasitoid Cotesia vanessae (Hymenoptera: Braconidae). Previously known from Eurasia and northern Africa, the first known recovery of C. vanessae in North America was in 2009. C. vanessae completed development in 34 species, of which three were known hosts (Noctuidae) and 31 (30 Noctuidae, 1 Nymphalidae) were not. Many of these noctuid species are economic pests. Parasitoid fitness was generally highest on species of Plusiinae (Noctuidae), measured as either percentage of successful parasitism, developmental time, or number and mass of F1 progeny. Closely related species were generally similar in their suitability as hosts. In some cases, parasitoid eggs or larvae were killed by the immune system of the parasitized host, but the host eventually failed to excrete food waste, did not pupate, and ultimately died. Such cases reached up to 100% mortality depending upon the lepidopteran species. The suitability of many species of noctuid pests as hosts for C. vanessae suggests that this parasitoid will become established widely throughout North America and may help to suppress populations of some pest species.

Biosynthesized silver nanoparticles (Ag NPs) from isolated actinomycetes strains and their impact on the black cutworm, Agrotis ipsilon.

Nanomaterials have been produced with the use of bio-nanotechnology, which is a low-cost approach. Currently, research is being conducted to determine whether actinomycetes isolated from Egyptian soil can biosynthesize Ag nanoparticles (Ag NPs) and characterized by using the following techniques: Transmission electron microscopy (TEM), Dynamic light scattering (DLS), Fourier transforms infrared (FT-IR), Energy-dispersive X-ray spectroscopy (EDX), UV-Vis spectroscopy and X-ray diffraction (XRD). The most promising actinomycetes isolate were identified, morphologically, biochemically, and molecularly. Streptomyces avermitilis Azhar A.4 was found to be able to reduce silver metal nanoparticles from silver nitrate in nine isolates collected from Egyptian soil. Toxicity of biosynthesized against 2nd and 4th larval instar of Agrotis ipsilon (Hufn.) (Lepidoptera: Noctuidae) was estimated. In addition, activity of certain vital antioxidant and detoxifying enzymes as well as midgut histology of treated larvae were also investigated. The results showed appositive correlations between larval mortality percentage (y) and bio-AgNPs concentrations (x) with excellent (R2). The 4th larval instar was more susceptible than 2nd larval instar with LC50 (with 95% confirmed limits) =8.61 (2.76-13.89) and 26.44(13.25-35.58) ppml-1, respectively of 5 days from treatment. The initial stages of biosynthesized AgNps exposure showed significant increases in carboxylesterase (CarE) and peroxidases (PODs) activity followed by significant suppression after 5 days pos-exposure. While protease activity was significantly decreased by increasing time post-exposure. Midgut histology showed abnormality and progressive damage by increasing time post exposure leading to complete destruction of midgut cells after 5 days from exposure. These results make biosynthesized AgNPs an appropriate alternative to chemical insecticide in A. ipsilon management.

An odorant receptor tuned to an attractive plant volatile vanillin in Spodoptera litura.

The insect olfaction plays crucial roles in many important behaviors, in which ORs are key determinants for signal transduction and the olfactory specificity. Spodoptera litura is a typical polyphagous pest, possessing a large repertoire of ORs tuning to broad range of plant odorants. However, the specific functions of those ORs remain mostly unknown. In this study, we functionally characterized one S. litura OR (OR51) that was highly expressed in the adult antennae. First, by using Xenopus oocyte expression and two-electrode voltage clamp recording system (XOE-TEVC), OR51 was found to be strongly and specifically responsive to vanillin (a volatile of S. litura host plants) among 77 tested odorants. Second, electroantennogram (EAG) and Y-tube behavioral experiment showed that vanillin elicited significant EAG response and attraction behavior especially of female adults. This female attraction was further confirmed by the oviposition experiment, in which the soybean plants treated with vanillin were significantly preferred by females for egg-laying. Third, 3D structural modelling and molecular docking were conducted to explore the interaction between OR51 and vanillin, which showed a high affinity (-4.46 kcal/mol) and three residues (Gln163, Phe164 and Ala305) forming hydrogen bonds with vanillin, supporting the specific binding of OR51 to vanillin. In addition, OR51 and its homologs from other seven noctuid species shared high amino acid identities (78-97%) and the same three hydrogen bond forming residues, suggesting a conserved function of the OR in these insects. Taken together, our study provides some new insights into the olfactory mechanisms of host plant finding and suggests potential applications of vanillin in S. litura control.

The Identification of a Quantative Trait Loci-Allele System of Antixenosis against the Common Cutworm (Spodoptera litura Fabricius) at the Seedling Stage in the Chinese Soybean Landrace Population.

Common cutworm (CCW) is an omnivorous insect causing severe yield losses in soybean crops. The seedling-stage mini-tray identification system with the damaged leaf percentage (DLP) as an indicator was used to evaluate antixenosis against CCW in the Chinese soybean landrace population (CSLRP) under three environments. Using the innovative restricted two-stage multi-locus genome-wide association study procedure (RTM-GWAS), 86 DLP QTLs with 243 alleles (2-11/QTL) were identified, including 66 main-effect loci with 203 alleles and 57 QTL-environment interaction loci with 172 alleles. Among the main-effect loci, 12 large-contribution loci (R2 ≥ 1%) explained 25.45% of the phenotypic variation (PV), and 54 small-contribution loci (R2 < 1%) explained 16.55% of the PV. This indicates that the CSLRP can be characterized with a DLP QTL-allele system complex that has not been found before, except for a few individual QTLs without alleles involved. From the DLP QTL-allele matrix, the recombination potentials expressed in the 25th percentile of the DLP of all possible crosses were predicted to be reduced by 41.5% as the maximum improvement and 14.2% as the maximum transgression, indicating great breeding potential in the antixenosis of the CSLRP. From the QTLs, 62 candidate genes were annotated, which were involved in eight biological function categories as a gene network of the DLP. Changing from susceptible to moderate plus resistant varieties in the CSLRP, 26 QTLs had 32 alleles involved, in which 19 genes were annotated from 25 QTL-alleles, including eight increased negative alleles on seven loci and 11 decreased positive alleles on 11 loci, showing the major genetic constitution changes for the antixenosis enhancement at the seedling stage in the CSLRP.

Biocontrol potential of cell suspensions and cell-free superntants of different Xenorhabdus and Photorhabdus bacteria against the different larval instars of Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae).

The black cutworm (BCW), Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), is one of the destructive cutworm species. Black cutworm is a highly polyphagous pest that feeds on more than 30 plants, many of which are of economic importance such as maize, sugar beet, and potato. The control of BCW larvae relies heavily on the application of synthetic insecticides which have a detrimental impact on human health and the natural environment. In addition, increasing insecticide resistance in many insect species requires a novel and sustainable approach to controlling insect pests. The endosymbionts of entomopathogenic nematodes (EPNs) (Xenorhabdus and Phorohabdus spp.) represent a newly emerging green approach to controlling a wide range of insect pests. In the current study, the oral and contact efficacy of cell suspension (4 × 107 cells ml-1) and cell-free supernatants of different symbiotic bacteria (X. nematophilai, X. bovienii, X. budapestensis, and P. luminescent subsp. kayaii) were evaluated against the mixed groups of 1st-2nd and 3rd-4th instars larvae of BCW under controlled conditions. The oral treatment of the cell suspension and cell-free supernatants resulted in higher mortality rates than contact treatments. In general, larval mortality was higher in the 1st-2nd instar larvae than in the 3rd-4th instar larvae. The highest (75%) mortality was obtained from the cell suspension of X. budapestensis. The results indicated that the oral formulations of the cell suspension and cell-free supernatants of bacterial strains may have a good control potential against the 1st-2nd larvae BCW. However, the efficacy of the cell suspension and cell-free supernatants of tested bacterial strains should be further evaluated under greenhouse and field conditions.

Soybean CALCIUM-DEPENDENT PROTEIN KINASE17 Positively Regulates Plant Resistance to Common Cutworm (Spodoptera litura Fabricius).

Soybean is frequently attacked by herbivorous pests throughout the growth period. Exploring anti-insect genes to improve insect resistance in soybean is an important soybean breeding goal. Here, we cloned and characterized the gene for a quantitative trait locus (QTL) related to insect resistance, Glyma.06g189600, which encodes CALCIUM-DEPENDENT PROTEIN KINASE17 (GmCDPK17) in soybean. The pairwise sequence alignment analysis revealed that the presumed protein of GmCDPK17 shares 52.06% similarity with that of GmCDPK38, a known negative regulatory gene of insect resistance in soybean. Ectopic expression of GmCDPK17 and GmCDPK38 restored the phenotypes of the Arabidopsis insect-susceptible mutant cpk10 and insect-resistant mutant cpk28, respectively. Moreover, transgenic hairy roots of the soybean cultivar Jack were generated by Agrobacterium-mediated transformation. Overexpression of GmCDPK17 increased soybean hairy root resistance to common cutworm (CCW), while RNA interference of the gene decreased soybean hairy root resistance to CCW. Sequencing data from the cultivated and wild soybeans were used to analyze the genetic diversity of GmCDPK17. This gene was subjected to domestication selection. Six and seven haplotypes (Haps) were identified in cultivated and wild soybeans, respectively. The resistance Hap1 is not widely used in cultivated soybeans and is mainly distributed at low latitudes. Accessions with resistance haplotypes of the GmCDPK17 and GmCDPK38 genes showed high resistance to CCW. Altogether, we revealed a novel positive regulatory insect resistance gene, GmCDPK17, which may further improve insect resistance in soybean.

Genome-Wide Association Studies Reveal Novel Loci for Herbivore Resistance in Wild Soybean (Glycine soja).

The production of soybean [Glycine max (L.) Merr.] is seriously threatened by various leaf-feeding insects, and wild soybean [Glycine soja Sieb. & Zucc.] has a greater resistance capacity and genetic diversity. In this study, a natural population consisting of 121 wild soybean accessions was used for detecting insect resistance genes. The larval weight (LW) of the common cutworm (CCW), the resistance level (RL) and the index of damaged leaf (IDL) were evaluated as resistance indicators to herbivores. An association synonymous SNP AX-94083016 located in the coding region of the respiratory burst oxidase gene GsRbohA1 was identified by genome-wide association study (GWAS) analyses. The overexpression of GsRbohA1 in soybean hairy roots enhanced resistance to CCW. One SNP in the promoter region cosegregated with AX-94083016 contributing to soybean resistance to CCW by altering GsRbohA1 gene expression and reactive oxygen species (ROS) accumulation. Two major haplotypes, GsRbohA1A and GsRbohA1G, were identified based on the SNP. The resistant haplotype GsRbohA1A predominates in wild soybeans, although it has been gradually lost in landraces and cultivars. The nucleotide diversity around GsRbohA1 is much lower in landraces and cultivars than in its ancestors. In conclusion, a new resistant haplotype, GsRbohA1A, was identified in wild soybean, which will be a valuable gene resource for soybean insect resistance breeding through introducing into improvement lines, and it offers a strategy for exploring resistance gene resources from its wild relatives.

Knockdown of GmVQ58 encoding a VQ motif-containing protein enhances soybean resistance to the common cutworm (Spodoptera litura Fabricius).

Plants have evolved complex defense mechanisms to withstand insect attack. Identification of plant endogenous insect resistance genes is of great significance for understanding plant-herbivore interactions and improving crop insect resistance. Soybean (Glycine max (L.) Merr.) is an important crop that is often attacked by the common cutworm (CCW) (Spodoptera litura Fabricius). In this study, based on our transcriptomic data, the gene GmVQ58, encoding a FxxxVQxxTG (VQ) motif-containing protein, was cloned and characterized. This gene showed the highest expression in the leaves and roots and was up-regulated significantly after CCW attack. Constitutive expression of GmVQ58 rescued the susceptibility of an Arabidopsis mutant to CCW, and interference of GmVQ58 in soybean hairy roots enhanced the resistance to CCW. Furthermore, GmVQ58 was localized to the nucleus and physically interacted with the transcription factor GmWRKY32. The expression of two defense-related genes, GmN:IFR and GmVSPß, was up-regulated in GmVQ58-RNAi lines. Additionally, the promoter region of GmVQ58 was likely selected during domestication, resulting in different expression patterns in cultivated soybeans relative to wild soybeans. These results suggest that silencing GmVQ58 confers soybean resistance to CCW.

Different binding properties of two general-odorant binding proteins in Athetis lepigone with sex pheromones, host plant volatiles and insecticides.

Athetis lepigone (Alep) is a polyphagous pest native to Europe and Asia that has experienced major outbreaks in the summer maize area of China since 2011 and has shown evidence of resistance to some insecticides. Insect olfaction is crucial for recognition of sex pheromones, host plant volatiles and even insecticides, in which two general-odorant binding proteins (GOBPs) play important roles. To elucidate the functions of GOBPs in A. lepigone, we first expressed the two AlepGOBP proteins in the E. coli expression system. Then, the results of fluorescence competitive binding assays demonstrated that the high binding affinity of AlepGOBP2 with sex pheromones [(Z)-7-dodecenyl acetate (Z7-12:Ac), Ki = 0.65 µM; (Z)-9-tetradecenyl acetate (Z9-14:Ac), Ki = 0.83 µM], two maize plant volatiles [Ocimene, Ki = 9.63 µM; (E)-ß-Farnesene, Ki = 4.76 µM] and two insecticides (Chlorpyrifos Ki =5.61 µM; Phoxim, Ki = 4.38 µM). However, AlepGOBP1 could only bind Ocimene (Ki = 13.0 µM) and two insecticides (Chlorpyrifos Ki =4.46 µM; Phoxim, Ki = 3.27 µM). These results clearly suggest that AlepGOBP1 and AlepGOBP2 differentiate among odorants and other ligands. The molecular docking results further revealed different key residues involved in the ligand binding of AlepGOBPs. In summary, this study provides a foundation for exploring the olfactory mechanism of A. lepigone and identified two potential target genes for the development of highly effective insecticides in the future.

RNA-Seq analysis reveals transcript diversity and active genes after common cutworm (Spodoptera litura Fabricius) attack in resistant and susceptible wild soybean lines.

BACKGROUND: Common cutworm (CCW) is highly responsible for destabilizing soybean productivity. Wild soybean is a resource used by breeders to discover elite defensive genes. RESULTS: The transcriptomes of two wild accessions (W11 and W99) with different resistance to CCW were analyzed at early- and late-induction time points. After induction, the susceptible accession W11 differentially expressed 1268 and 508 genes at the early and late time points, respectively. Compared with W11, the resistant accession W99 differentially expressed 1270 genes at the early time point and many more genes (2308) at the late time point. In total, 3836 non-redundant genes were identified in both lines. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed that the differentially expressed genes (DEGs) in W99 at the late time point were mostly associated with specific processes and pathways. Among the non-redundant genes, 146 genes were commonly up-regulated in the treatment condition compared with the control condition at the early- and late-induction time points in both accessions used in this experiment. Approximately 40% of the common DEGs were related to secondary metabolism, disease resistance, and signal transduction based on their putative function. Excluding the common DEGs, W99 expressed more unique DEGs than W11. Further analysis of the 3836 DEGs revealed that the induction of CCW not only up-regulated defense-related genes, including 37 jasmonic acid (JA)-related genes, 171 plant-pathogen-related genes, and 17 genes encoding protease inhibitors, but also down-regulated growth-related genes, including 35 photosynthesis-related genes, 48 nutrition metabolism genes, and 28 auxin metabolism genes. Therefore, representative defense-related and growth-related genes were chosen for binding site prediction via co-expression of transcription factors (TFs) and spatial expression pattern analyses. In total, 53 binding sites of 28 TFs were identified based on 3 defense-related genes and 3 growth-related genes. Phosphate transporter PT1, which is a representative growth-related gene, was transformed into soybean, and the transgenic soybean plants were susceptible to CCW. CONCLUSIONS: In summary, we described transcriptome reprograming after herbivore induction in wild soybean, identified the susceptibility of growth-related genes, and provided new resources for the breeding of herbivore-resistant cultivated soybeans.

Evaluation of the resistance effect of QTLs derived from wild soybean (Glycine soja) to common cutworm (Spodoptera litura Fabricius).

Common cutworm (CCW) is a serious herbivorous insect pest of soybean. Previously, we conducted an antixenosis bioassay (measuring feeding preference) with CCW using recombinant inbred lines (RILs) derived from a cross between a wild soybean (Glycine soja) collected in Hiroshima prefecture (JP110755) and the leading cultivar, Fukuyutaka. The analysis revealed quantitative trait loci (QTLs) for antixenosis resistance, qRslx3 and qRslx4. In the present study we developed another RIL population using Fukuyutaka and a different G. soja, collected in Kumamoto prefecture (G406). An analysis revealed an antixenosis resistance QTL on chromosome 7, and the resistant allele of the QTL was derived from G406. The chromosomal position of the QTL was almost the same as that of CCW-2, a previously-reported antibiosis resistance QTL for CCW, detected in a F2 population derived from a cross between Fukuyutaka and a resistant cultivar Himeshirazu. These QTLs could be the same locus; however, G406 and Himeshirazu are likely to possess different alleles, because Himeshirazu allele exhibits no antixenosis effect. We expect that pyramiding of the resistance QTLs derived from G. soja will contribute to the development of CCW resistant cultivars.

A Δ9 desaturase (SlitDes11) is associated with the biosynthesis of ester sex pheromone components in Spodoptera litura.

Sex pheromone biosynthesis in moths relies on the activity of multiple enzymes, including Δ9 desaturase, which plays an important role in catalyzing desaturation at the Δ9 position of the carbon chain. However, the physiological function of moth Δ9 desaturase has not been elucidated in vivo. In this study, we used the CRISPR/Cas9 system to knockout the Δ9 desaturase gene (SlitDes11) of Spodoptera litura to analyze its role in sex pheromone biosynthesis. First, through the direct injection of SlitDes11-single guide RNA (sgRNA)/Cas9 messenger RNA into newly laid eggs, gene editing was induced in around 30% of eggs 24 h after injection and was induced in 20.8% of the resulting adult moths. Second, using a sibling-crossing strategy, insects with mutant SlitDes11 (bearing a premature stop codon) were selected, and homozygous mutants were obtained in the G5 generation. Third, pheromone gland extracts of adult female homozygous SlitDes11 mutants were analyzed using Gas chromatography (GC). The results showed that titers of all three ester sex pheromone components; Z9, E11-14:Ac, Z9,E12-14:Ac, and Z9-14:Ac; were reduced by 62.40%, 78.50%, and 72.50%, respectively. This study provides the first direct evidence for the role of SlitDes11 in sex pheromone biosynthesis in S. litura, and indicates the gene could be as potential target to disrupt sexual communication in S. litura for developing a new pollution-free insecticide.

Life-History Parameters of Striacosta albicosta (Lepidoptera: Noctuidae) Under Laboratory Conditions.

Striacosta albicosta (Smith) is a key pest of maize and dry beans in North America. It has expanded its distribution from the western Great Plains of the United States to the Great Lakes region in the United States and Canada. There has been limited research on the baseline biological aspects of this insect under controlled conditions. The objective of this study was to detail the biological parameters of S. albicosta feeding on an artificial diet under laboratory conditions. Overall survival from neonate to adult at 26.6 ± 1°C was 36.72% and the total developmental time was approximately 110 d. Survival of the egg, larval, prepupal, and pupal stages were 75.71, 98.50, 51.78, and 95.10%, respectively. Average duration of the egg, larval, prepupal, and pupal stages was 4.64, 28.20, 41.50, and 25.91 d, respectively. During the larval stage, 92.50% of larvae developed through seven instars and the remaining through six instars. Larvae that developed through six and seven instars exhibited a mean growth ratio of 1.60 and 1.47, respectively; however, there was no difference in pupal weight. Eggs laid by field-mated moths showed a fertility of 75.71%, compared with 4.18% from laboratory-reared moths. These data suggest that S. albicosta develop primarily through seven instars and the most vulnerable developmental stage is the prepupa. Laboratory conditions strongly affected fertility success. Information presented here greatly expands our understanding of S. albicosta biology, which can be used to improve the efficiency of laboratory bioassays and management techniques for this critical crop pest.

α-Ionone, an Apocarotenoid, Induces Plant Resistance to Western Flower Thrips, Frankliniella occidentalis, Independently of Jasmonic Acid.

Apocarotenoids, such as ß-cyclocitral, α-ionone, ß-ionone, and loliolide, are derived from carotenes via chemical or enzymatic processes. Recent studies revealed that ß-cyclocitral and loliolide play an important role in various aspects of plant physiology, such as stress responses, plant growth, and herbivore resistance. However, information on the physiological role of α-ionone is limited. We herein investigated the effects of α-ionone on plant protection against herbivore attacks. The pretreatment of whole tomato (Solanum lycopersicum) plants with α-ionone vapor decreased the survival rate of western flower thrips (Frankliniella occidentalis) without exhibiting insecticidal activity. Exogenous α-ionone enhanced the expression of defense-related genes, such as basic ß-1,3-glucanase and basic chitinase genes, in tomato leaves, but not that of jasmonic acid (JA)- or loliolide-responsive genes. The pretreatment with α-ionone markedly decreased egg deposition by western flower thrips in the JA-insensitive Arabidopsis (Arabidopsis thaliana) mutant coi1-1. We also found that common cutworm (Spodoptera litura) larvae fed on α-ionone-treated tomato plants exhibited a reduction in weight. These results suggest that α-ionone induces plant resistance to western flower thrips through a different mode of action from that of JA and loliolide.

Genetic and phenotypic comparisons of viral genotypes from two nucleopolyhedroviruses interacting with a common host species, Spodoptera litura (Lepidoptera: Noctuidae).

Nucleopolyhedroviruses (NPVs) are known to be highly variable, both genetically and phenotypically, at several scales such as different geographic locations or a single host. A previous study using several geographic isolates indicated that two types of NPV, Spodoptera littoralis NPV (SpliNPV) and S. litura NPV (SpltNPV) types, were isolated from the common cutworm, Spodoptera litura (Fabricius), a polyphagous insect that causes serious damage to many forage crops and vegetables. That study also indicated that the SpliNPV type was widely distributed in Japan. Here, we investigated the genotypic and phenotypic variation of cloned NPVs that infect S. litura; such variation is an important resource for biological control agents, and may represent the genetic diversity of an NPV species. Eighteen genotypically distinct NPVs were cloned from four field-collected NPV isolates using an in vivo cloning technique. They were divided into two virus types according to the similarity of banding patterns of DNA fragments generated by restriction endonucleases, and Southern hybridization analysis. Partial polyhedrin gene sequences revealed that the two types corresponded to SpliNPV and SpltNPV. Bioassays seem to suggest that the SpliNPV virus type was, overall, more infectious and killed S. litura larvae faster, but yielded fewer viral occlusion bodies, than the SpltNPV type. These data provide a basis for explaining the distribution pattern of SpliNPV and SpltNPV types in S. litura populations in Japan.

Toxicity of Alangium salvifolium Wang chemical constituents against the tobacco cutworm Spodoptera litura Fab.

Widespread use of synthetic pesticides has resulted in the development of insecticide-resistant populations of pests and harmful effects on human health and the environment. There is a need to identify alternative pest management strategies to reduce our reliance on conventional chemical pesticides. In recent years the use of botanical pesticides for protecting crops from insect pests has assumed greater importance. Methanol extract of Alangium salvifolium (L.f.) Wang has potential insecticidal activity against Spodoptera litura Fab. The active fractions were identified through chromatographic techniques as F-IV (Rf value=0.45) and F-VI (Rf value=0.63) and were subjected to GC-MS (GCMATE II). Fifty, 100 and 200ppm of active fractions were applied to fourth instar larvae and the mortality increased with higher concentrations. Relative consumption rate, relative growth rate, efficiency of conversion of ingested food and efficiency of conversion of digested food values all decreased in treated larvae, but approximate digestibility rate increased after treatment. The hydrolytic enzymes, such as acid phosphatase, alkaline phosphatase and the glycolytic enzyme lactate dehydrogenase were inhibited in treated larvae compared with controls. The histopathology study revealed that the epithelial columnar cells were enlarged, completely atrophied; intercellular spaces were swollen, and also noted a cytoplasmic ooze of cell material that mixed with food column. The present study clearly showed the active fractions from A. salvifolium as potential botanicals to control the larvae of S. litura. This is the first report for nutritional indices, enzymatic activities and histological effects of A. salvifolium chemical constituents against S. litura. Thus probably, this will be used as an alternative for synthetic pesticides against the polyphagous pest like S. litura.

Unclassified glutathione-S-transferase AiGSTu1 confers chlorantraniliprole tolerance in Agrotis ipsilon.

BACKGROUND: Chlorantraniliprole (CAP) is a diamide insecticide with high efficacy against many pest insects, including the black cutworm, Agrotis ipsilon. Agrotis ipsilon is a serious pest causing significant yield losses in crops. Glutathione-S-transferases (GSTs) belong to a family of metabolic enzymes that can detoxify a wide range of pesticides. However, little is known about the functions of GSTs in CAP tolerance in A. ipsilon. RESULTS: A cDNA sequence (designated AiGSTu1) encoding an unclassified GST was identified from A. ipsilon. AiGSTu1 is highly expressed during the 3rd -instar larval and the pupal stages. Most of the mRNA transcripts were found in larval Malpighian tubules. Exposure to CAP strongly enhanced AiGSTu1 expression, GST activity, hydrogen peroxide (H2 O2 ) and malondialdehyde levels in larvae. H2 O2 treatment upregulated the transcription level of AiGSTu1, suggesting that CAP-induced oxidative stress may activate AiGSTu1 expression. The activity of recombinant AiGSTu1 was inhibited by CAP in a dose-dependent manner. Metabolism assay results demonstrated that AiGSTu1 is capable of depleting CAP. Overexpression of AiGSTu1 enhanced the tolerance of Escherichia coli cells to H2 O2 and the oxidative stress inducer, cumene hydroperoxide. Silencing of AiGSTu1 by RNA interference increased the susceptibility of A. ipsilon larvae to CAP. CONCLUSION: The findings of this study provide valuable insights into the potential role of AiGSTu1 in CAP detoxification and will improve our understanding of CAP tolerance in A. ipsilon. © 2023 Society of Chemical Industry.