Microorganisms symbiotic with insects, whether permanently or temporarily, play a crucial role in the nutrition, development, reproduction, defence, and metamorphosis regulation. In some Lepidoptera, oviposition-deterrent pheromones (ODPs) on egg surface were used by pregnant females to modify the behaviour of conspecifics to avoid excessive competition for limited resources. In this study, we constructed four different Spodoptera litura groups, including, OH, OA, SH, and OA, which either feed on different hosts or grow in different environments. The 16S rDNA libraries of microbes from the egg surface of the four groups were constructed and sequenced. According to alpha and beta diversity indices, the microbes in environments and diets considerably influenced the richness, diversity, and community compositions of the microbiota on egg surfaces. The quantity of the main ODP components and the corresponding oviposition-deterrent activity among four groups were significantly differed among the four groups. The result of this study revealed that altering of microbes in environments or diets considerably changed the contents of ODP and oviposition-deterrent activity. As ODPs impart oviposition-deterrent activity towards closely related species, the findings of this study suggest that we should pay more attention to the role of symbiotic microorganisms in changing the ability of insects, especially sympatric species, to occupy the optimal niche when developing novel pest-control strategies.

The influences of different plants on herbivores have recently attracted research interest; however, little is known regarding the effects of wild, local and cultivated varieties of the same plant from the same origin on herbivores. This study aimed to examine the effects of different tobacco varieties from the same origin on the oviposition preference and offspring performance of Spodoptera litura. We selected two wild (‘Bishan wild tobacco’ and ‘Badan wild tobacco’), two local (‘Liangqiao sun-cured tobacco’ and ‘Shuangguan sun-cured tobacco’) and two cultivated (‘Xiangyan No. 5’ and ‘Cunsanpi’) tobacco varieties from Hunan Province, China. We found that female S. litura varied in oviposition preferences across the tobacco varieties. They preferred to lay eggs on the cultivated varieties, followed by the local varieties, with the wild varieties being the least preferred. Furthermore, different tobacco varieties significantly influenced the life history parameters of S. litura. Survival rate, pupal weight, emergence rate and adult dry weight decreased in the following order: cultivated varieties > local varieties > wild varieties. Conversely, the pupal stage and development period decreased in the following order: wild varieties > local varieties > cultivated varieties. Therefore, we conclude that wild tobacco varieties have higher resistance to S. litura than cultivated and local varieties, reflecting the evolutionary advantages of wild tobacco varieties.

Spodoptera litura is an important pest that causes significant economic damage to numerous crops worldwide. Sex pheromones (SPs) mediate sexual communication in S. litura and show a characteristic degree of rhythmic activity, occurring mainly during the scotophase; however, the specific regulatory mechanisms remain unclear. Here, we employed a genome-wide analysis to identify eight candidate circadian clock genes in S. litura. Sequence characteristics and expression patterns were analyzed. Our results demonstrated that some circadian clock genes might regulate the biosynthesis and perception of SPs by regulating the rhythmic expression of SP biosynthesis-related genes and SP perception-related genes. Interestingly, all potential genes exhibited peak expression in the scotophase, consistent with the SP could mediate courtship and mating behavior in S. litura. Our findings are helpful in elucidating the molecular mechanism by which circadian clock genes regulate sexual communication in S. litura.

The tobacco cutworm Spodoptera litura (Lepidoptera: Noctuidae) is a polyphagous pest with a highly selective and sensitive chemosensory system involved in complex physiological behaviors such as searching for food sources, feeding, courtship, and oviposition. However, effective management strategies for controlling the insect pest populations under threshold levels are lacking. Therefore, there is an urgent need to formulate eco-friendly pest control strategies based on the disruption of the insect chemosensory system. In this study, we identified 158 putative chemosensory genes based on transcriptomic and genomic data for S. litura, including 45 odorant-binding proteins (OBPs, nine were new), 23 chemosensory proteins (CSPs), 60 odorant receptors (ORs, three were new), and 30 gustatory receptors (GRs, three were new), a number higher than those reported by previous transcriptome studies. Subsequently, we constructed phylogenetic trees based on these genes in moths and analyzed the dynamic expression of various genes in head capsules across larval instars using quantitative real-time polymerase chain reaction. Nine genes–SlitOBP8, SlitOBP9, SlitOBP25, SlitCSP1, SlitCSP7, SlitCSP18, SlitOR34, SlitGR240, and SlitGR242–were highly expressed in the heads of 3- to 5-day-old S. litura larvae. The genes differentially expressed in olfactory organs during larval development might play crucial roles in the chemosensory system of S. litura larvae. Our findings substantially expand the gene inventory for S. litura and present potential target genes for further studies on larval feeding in S. litura.

Spodoptera litura is a widespread polyphagous insect pest that can develop resistance and cross-resistance to insecticides, making it difficult to control. Insecticide exposure has previously been linked with induction of specific olfactory-related proteins, including some chemosensory proteins (CSPs) and odorant-binding proteins (OPBs), which may disrupt detection of environmental factors and reduce fitness. However, functional evidence supporting insecticide and OBPs/CSPs mediation remains unknown. Here we fed male S. litura moths with sucrose water containing one of three insecticides, chlorpyrifos, emamectin benzoate or fipronil, and used real-time quantitative polymerase chain reaction and RNAi to investigate OBPs and CSPs expression and their correlations with survival. Chlorpyrifos and emamectin benzoate increased expression of 78% of OBPs, plus 63 and 56% of CSP genes, respectively, indicating a major impact on these gene families. RNAi knockdown of SlituCSP18, followed by feeding with chlorpyrifos or fipronil, decreased survival rates of male moths significantly compared with controls. Survival rate also decreased significantly with the downregulation of SlituOBP9 followed by feeding with chlorpyrifos. Thus, although these three insecticides had different effects on OBP and CSP gene expression, we hypothesize that SlituOBPs and SlituCSPs might mediate their effects by increasing their expression levels to improve survival. Moreover, the differential response of S. litura male moths to the three insecticides indicated the potential specificity of chlorpyrifos affect SlituCSP18 and SlituOBP9 expression.

The present study was conducted to appraise the ontogenic radio-sensitivity of a serious tropical pest, Spodoptera litura (Fabr.). The molecular responses pertaining to the phenoloxidase (PO) pathway and an anti-oxidant defense mechanism were evaluated in order to understand its implication in pest control at pre-harvest and post-harvest intervals. Irradiation exhibited an inverse relationship with age with respect to impact on developmental and transcriptional responses. Transcript abundance of PO cascade enzymes, prophenoloxidase (slppo-2), its activating enzyme (slppae-1) and free-radical scavenging enzymes, superoxide dismutase (slsod) and catalase (slcat) was evaluated upon gamma irradiation alone and the dual-stress of radiation plus microbial challenge. The slppo-2, slppae-1, slsod and slcat transcripts were significantly up-regulated in F1 L6 larvae (6th-instar) resulting from 100 Gy sub-sterilized male adults and unirradiated female moths. The extent of upregulation was relatively higher in comparison with L6 survivors (6th-instar larvae) developed from irradiated neonates (L1) treated with 100 Gy. Upon Photorhabdus challenge, the transcripts were down-regulated in irradiated L1 suggesting increased larval susceptibility to bacterial infections. Radioresistance increased with the age of the insect, and molecular responses (transcript abundance) of insect defense mechanism were less influenced when older age (F1 progeny) were irradiated. These findings will help to optimize the gamma dose to be employed in inherited sterility technique for (pre-harvest) pest suppression and (post-harvest) phytosanitation and quarantine, and suggest compatible integration of biorational tactics including nuclear technology.

Addition of antibiotics to artificial diets of insects is a key component in the rearing of insects in the laboratory. In the present study an antimicrobial agent, streptomycin sulphate was tested for its influence on survival and fitness of Spodoptera litura (Fabricus) (Lepidoptera: Noctuidae) as well as its gut microbial diversity. The antibiotic did not adversely affect the survival of S. litura. Faster growth of larvae was recorded on diet amended with different concentrations of streptomycin sulphate (0.03, 0.07 and 0.15%) as compared to diet without streptomycin sulphate. The overall activity of various digestives enzymes increased on S+ diet while the activity of detoxifying enzymes significantly decreased. In addition, alteration in microbial diversity was found in the gut of S. litura larvae fed on diet supplemented with antibiotic (S+) and without antibiotic (S−).

Snellenius manilae (Ashmead) and Meteorus pulchricornis (Wesmael) (Hymenoptera: Braconidae) are larval endoparasitoids of Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae). Both species preferentially parasitize early-instar S. litura and occupy similar ecological niches. Therefore, competition between the two species may occur. In this study, intrinsic competition and cage experiments were conducted to discuss the interactions between S. manilae and M. pulchricornis. The results indicated that in intrinsic competition, M. pulchricornis was always the dominant species. In cage experiments, when the total number of parasitoids was four, the parasitism rates following the release of one species were significantly higher than the release of two species simultaneously. In addition, parasitism rate of eight M. pulchricornis was also significantly higher than the parasitism rate of the treatment released four S. manilae and four M. pulchricornis simultaneously. Therefore, competition occurs between S. manilae and M. pulchricornis, and M. pulchricornis is typically the superior of the two species. The use of M. pulchricornis as a biological agent for S. litura should be considered.

Combinations of Bacillus thuringiensis subsp. kurstaki (Btk) and some plant extracts, namely Acacia arabica (Lam.) Willd., Annona squamosa L., Datura stramonium L., Eucalyptus globulus (Labille), Ipomoea carnea Jacq., Lantana camara L., Nicotiana tabacum L. and Pongamia pinnata (L.) Pierre, were studied for their efficacy and synergistic activity against Spodoptera litura (Fab.) larvae. D.stramonium seed extract fortified with Btk recorded highest mean larval mortality of 74.7%, followed by 61% with A. arabica seed extract, 54.4% with N. tabacum leaf extract and 53% with A. squamosa seed extract. Combination of A. arabica seed extract at 25% concentration and Btk resulted in considerable mortality within 24 h. All Btk fortified extracts caused higher larval mortality than with Btk alone, indicating synergistic action except for the A. arabica leaf extract and Btk combination, which showed antagonistic effect.

We selected 50 crystal (Cry) toxins specific to lepidopteran insects. Phylogenetic relationships of whole toxins as well as individual domains revealed that Cry1Jb and Cry1Ac are distantly placed but related to reduced cross-resistance in Helicoverpa armigera. Multiple alignments of Cry1Jb and Cry1Ac amino acid sequences showed significant differences in the composition and length of the loops of domains II and III. This was further confirmed by the superpositioning of 3D structures. cry1Jb and cry1Ac genes cloned in expression vectors were overexpressed in Escherichia coli Castellani & Chalmers, and proteins were harvested. Insect bioassays revealed that the wild-type Cry1Jb and Cry1Ac proteins showed differential specificity to H. armigera (Hübner), Spodoptera litura (Fab.) and Earias vittella F. Chimeric genes were constructed by exchanging the domains between cry1Jb and cry1Ac and overexpressed in BL21(DE3). Substitution of domain III of Cry1Jb with that of Cry1Ac enhanced its activity against H. armigera 7.8-fold. Bioassay of the parental and chimeric toxins against S. litura revealed no activity even at the discriminative dose of 100 μg/ml, but a significant difference in growth was observed. The present results, along with previous domain swapping experiments, suggest that protein engineering not only reveals the mechanism by which endotoxins work, but also generates novel toxins with enhanced toxicity/broader specificity.

The effects of pH, sunlight and rearing temperature on infectivity of nucleopolyhedrovirus (SpltNPV) were investigated against larvae of Spodoptera litura (Fabricius). The infectivity of virus increased with increased larval rearing temperature. The LT50 values were significantly reduced from 24 to 6 days when rearing temperature was increased from 20 to 35 °C. The pH of the substrate significantly affected the infectivity of the virus. The LT50 values increased to about fourfold when the pH of suspension was raised from 7 to 11. Infectivity of the virus was more adversely affected in alkaline than in acidic suspension. Sunlight also deleteriously affected infectivity of the virus. The LT50 values increased with the increase in the period of sunlight exposure. A complete viral inactivation occurred after 12 h of direct sunlight exposure.

A microsporidium was isolated from larvae of Spodoptera litura (Fabricius) collected from Tamil Nadu, India. This microsporidian species is monomorphic, disporous and develops in direct contact with the cytoplasm of the host cell. The nuclear configuration of merogonic and sporogonic stages was diplokaryotic. The merogonic proliferative stage was unusual that normal development with 1, 2 and 4 binucleated forms were common, while large multinucleate meronts containing 8 and 12 small compact horseshoe-like diplokaryotic nuclei were also observed. The fresh spores were typically ovocylindrical in shape, with a mean size of 3·91×1·91 μm and the polar filament length was ~90 μm. Infection was systemic with mature spores produced in the midgut, nervous tissue, muscles, labial glands, gonads, tracheae, epidermis, Malpighian tubules and, most extensively, fat body tissues. The new isolate was highly pathogenic to S. litura larvae. Host specificity tests performed on 37 non-target hosts of 5 different insect orders revealed that the new isolate is pathogenic only to lepidopteran insects. We sequenced the 16S small subunit rRNA (SSU rRNA) gene of the isolate and compared it with 72 non-redundant microsporidian sequences from the GenBank. Based on the light microscopic studies and phylogenetic analyses, the new isolate is assigned to the genus Nosema. Significant differences in the SSU rRNA sequence were identified when compared with the type species Nosema bombycis and other closely related species viz., Nosema spodopterae. Structural differences were also observed in the 16S SSU rRNA between the new isolate and the two above-mentioned microsporidian pathogens. We conclude that the microsporidian isolate reported here is distinctly different from the other known species and is likely to be a new species.

An artificial diet for mass rearing of the tobacco caterpillar Spodoptera litura (Fabricius) from the neonate to adult stage was developed at 27±1°C, 65±5% RH and 16:8 h scoto/photo-phase regime. The diet ingredients consisted of wheat germ (26.0 g), kidney bean flour (51.3 g), chickpea flour (56.0 g), yeast powder (31.6 g), casein (15.2 g), l-ascorbic acid (3.2 g), cholesterol (0.5 g), two multivitamin multimineral capsules, one vitamin E capsule, ABDEC drops (2 ml), castor oil (1 ml), methyl-p-hydroxybenzoate (1.8 g), sorbic acid (1.3 g), streptomycin sulphate (0.25 g), formaldehyde solution (2 ml), agar–agar (16.4 g) and 820 ml distilled water. The new artificial diet successfully supported the growth and development for more than ten generations with enhanced reproductive potential. The mean biological parameters based on rearing of ten continuous generations showed higher pupation (89.2%), emergence (97.2%), survival (86.6%) and fecundity (2486.2 eggs) as compared to the most preferred natural food, castor leaf whose respective recorded values are 80, 75, 60% and 480 eggs. The cost of 1 l of the diet was approximately US$ 2.00 only, on which 200 neonate larvae can be reared. The artificial diet developed is suitable for mass rearing S. litura successfully throughout the year for laboratory and field experimentation as well as for commercial production of nuclear polyhedrosis viruses (Sl NPV).

Spodoptera litura (F.) (Lepidoptera: Noctuidae) is a serious pest of several crops in India and other countries. Prothoracicotropic hormone (PTTH) release from the brain of penultimate fifth instars of the noctuid was followed in the laboratory and a spiracle index developed to assess PTTH action. PTTH release from the brain occurred during the scotophase at 4.30 h of day 1 (AZT). Ecdysone secreted in the prothoracic gland was released from the 7th hour of day 1 (AZT), reaching a maximum in the haemolymph on the 9th hour of day 1 (AZT). The prothoracic gland showed three distinct phases, namely growth, synthesis and release. Injected PTTH of 5th instar brain extract induced larval moulting in head-ligated larvae. Partially purified PTTH from 6th-instar S. litura brain induced moulting in 6th instars. Abdomen-ligated penultimate instars administered β-ecdysone showed variable moulting response. The phenomenon of hormonal interaction with development of the penultimate instar of S. litura is discussed.

The development, life table and intrinsic rate of natural increase of the reduviid predator Rhynocoris fuscipes (Fabricius) on Spodoptera litura Fab., Earias vittella Fab. and Corcyra cephalonica Stainton under laboratory conditions were constructed. Fecundity, longevity and minimum developmental period of the predator were dependent on the prey diet and decreased in the order: S. litura, E. vittella and C. cephalonica. The mean number of female progeny produced over the entire period (the gross reproductive rate) was 43.0, 37.5 and 32.5 on S. litura, E. vittella and C. cephalonica respectively. The net reproductive rate (Ro), intrinsic rate of increase (rc) and weekly multiplication rate were also highest on S. litura followed by E. vittella and C. cephalonica.

The production of the nucleopolyhedrovirus (NPV) of the tobacco cutworm, Spodoptera litura was studied to determine the optimum conditions for in vivo production using homologous host larvae reared on artificial diet. We compared NPV production in 7, 8, 9 and 10-day-old larvae and found that 9-day-old larvae gave the highest virus yield. The optimum dose was determined by infecting 9-day old larvae (weighing 125–155 mg) with a range of doses. The maximum NPV production achieved was 3.91 × 109 polyhedral inclusion bodies (PIBs) per larva using an inoculum of 4.8 × 106 PIBs. A 7-day incubation period was found to be optimum for NPV production. Moribund larvae were found to be more suitable for handling and for the mass production of virus than dead ones.

The effects of neem seed kernel extract (NSKE), neem oil (NO) and nucleopolyhedrovirus (NPV) of the Spodoptera litura Fabricius (Lepidoptera: Noctuidae) on the mortality, feeding and development of S. litura were evaluated in the laboratory. Dose-dependent mortality was evident after NPV treatment, which also depended on larval age at treatment. Upon combining NPV with neem products, the percentage mortality was enhanced threefold even at reduced concentrations. The NSKE and NO significantly inhibited the growth and development of S. litura, and extended the duration of its larval and pupal stages. Oviposition period, adult longevity and fecundity were significantly reduced following treatment with neem products and with neem + NPV treatment. The efficiencies of conversion of ingested food (ECI) and digested food (ECD) were also significantly decreased by treatment with NSKE, NO and NPV, either alone or in combination.

Evaluations of toxicity, antifeedant, growth-regulatory activity of the methanol extract of Gliricidia sepium leaves were carried out against Dysdercus koenigii Fabr. (Heteroptera: Pyrrhocoridae), Achaea Janata Linn, and Spodoptera litura Fab., (both Lepidoptera: Noctuidae). Topical application of 100-300 μg of the extract to different stages of the test insects resulted in dose-dependent effects including larval-pupal intermediates and deformed adults. The highest tested dosage (300 μg/ μl) resulted in reduced survival rate and morphological abnormalities. At certain doses a strong antifeedant activity was evident against the lepidopteran insects.

The ecdysteroid agonist KK-42 [l-benzyl-5(E)-2,6- dimethyl-1,5-heptadienyl] imidazole adversely affected the growth of Spodoptera litura and caused a delay in pupation. The compound also caused adult deformation, increased sex ratio in favour of males, and adversely affected the fecundity and fertility of emergent females. The ecdysteroid 20-hydroxyecdysone, identified by HPLC, was present in treated and control insects. Ecdysone was also detected in only the insects treated with 1 μg/larva KK-42.

Bioenergetica, efficiencies of energy transfer and growth of Spodoptera litura larvae reared on mulberry, cabbage, castor and wild hempweed leaves were determined using calorimetrie procedures. Energy contents (J mg−1 ash-free tissue) of castor leaves were higher than those of cabbage, mulberry and wild hempweed. Fresh leaves had slightly higher energy contents than those left over. The rates of ingestion and egestion of food energy increased linearly with the age of larvae. Energy consumption decreased exponentially with larval age, and this decreasing trend showed an intermediate rise at the fifth instar. Assimilation efficiencies were highest and lowest for the first and sixth instar larvae respectively. Efficiencies of gross and net larval production showed linear increases with instar stage, except at the fifth instar which showed dramatic declines in conversion of ingested and assimilated energy into insect biomass. Efficiencies of gross and net productions during early first to third instars were low, but those during the sixth instar were high. The pathways of energy transfer indicated that an average of 58.5% of the total energy ingested was lost directly through faeces, respiration and exuviae, while about 42.2% of the total energy ingested was actually converted into growth of an S. litura larva.