Multifaceted interactions between host ESCRT-III and budded virus-related proteins involved in entry and egress of the baculovirus Autographa californica multiple nucleopolyhedrovirus.

The endosomal sorting complex required for transport (ESCRT) is a conserved protein machine mediating membrane remodeling and scission. In the context of viral infection, different components of the ESCRT-III complex, which serve as the core machinery to catalyze membrane fission, are involved in diverse viruses' entry, replication, and/or budding. However, the interplay between ESCRT-III and viral factors in the virus life cycle, especially for that of large enveloped DNA viruses, is largely unknown. Recently, the ESCRT-III components Vps2B, Vps20, Vps24, Snf7, Vps46, and Vps60 were determined for entry and/or egress of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV). Here, we identified the final three ESCRT-III components Chm7, Ist1, and Vps2A of Spodoptera frugiperda. Overexpression of the dominant-negative forms of these proteins or RNAi downregulation of their transcripts significantly reduced infectious budded viruses (BVs) production of AcMNPV. Quantitative PCR together with confocal and transmission electron microscopy analysis revealed that these proteins were required for internalization and trafficking of BV during entry and egress of nucleocapsids. In infected Sf9 cells, nine ESCRT-III components were distributed on the nuclear envelope and plasma membrane, and except for Chm7, the other components were also localized to the intranuclear ring zone. Y2H and BiFC analysis revealed that 42 out of 64 BV-related proteins including 35 BV structural proteins and 7 non-BV structural proteins interacted with single or multiple ESCRT-III components. By further mapping the interactome of 64 BV-related proteins, we established the interaction networks of ESCRT-III and the viral protein complexes involved in BV entry and egress.IMPORTANCEFrom archaea to eukaryotes, the endosomal sorting complex required for transport (ESCRT)-III complex is hijacked by many enveloped and nonenveloped DNA or RNA viruses for efficient replication. However, the mechanism of ESCRT-III recruitment, especially for that of large enveloped DNA viruses, remains elusive. Recently, we found the ESCRT-III components Vps2B, Vps20, Vps24, Snf7, Vps46, and Vps60 are necessary for the entry and/or egress of budded viruses (BVs) of Autographa californica multiple nucleopolyhedrovirus. Here, we demonstrated that the other three ESCRT-III components Chm7, Ist1, and Vps2A play similar roles in BV infection. By determining the subcellular localization of ESCRT-III components in infected cells and mapping the interaction of nine ESCRT-III components and 64 BV-related proteins, we built the interaction networks of ESCRT-III and the viral protein complexes involved in BV entry and egress. These studies provide a fundamental basis for understanding the mechanism of the ESCRT-mediated membrane remodeling for replication of baculoviruses.

Plant defenses interact with insect enteric bacteria by initiating a leaky gut syndrome.

Plants produce suites of defenses that can collectively deter and reduce herbivory. Many defenses target the insect digestive system, with some altering the protective peritrophic matrix (PM) and causing increased permeability. The PM is responsible for multiple digestive functions, including reducing infections from potential pathogenic microbes. In our study, we developed axenic and gnotobiotic methods for fall armyworm (Spodoptera frugiperda) and tested how particular members present in the gut community influence interactions with plant defenses that can alter PM permeability. We observed interactions between gut bacteria with plant resistance. Axenic insects grew more but displayed lower immune-based responses compared with those possessing Enterococcus, Klebsiella, and Enterobacter isolates from field-collected larvae. While gut bacteria reduced performance of larvae fed on plants, none of the isolates produced mortality when injected directly into the hemocoel. Our results strongly suggest that plant physical and chemical defenses not only act directly upon the insect, but also have some interplay with the herbivore's microbiome. Combined direct and indirect, microbe-mediated assaults by maize defenses on the fall armyworm on the insect digestive and immune system reduced growth and elevated mortality in these insects. These results imply that plant-insect interactions should be considered in the context of potential mediation by the insect gut microbiome.

Curcumin induces autophagic cell death in Spodoptera frugiperda cells.

The increasing interest in the role of autophagy (type II cell death) in the regulation of insect toxicology has propelled study of investigating autophagic cell death pathways. Turmeric, the rhizome of the herb Curcuma longa (Mañjal in Tamil, India and Jianghuáng in Chinese) have been traditionally used for the pest control either alone or combination with other botanical pesticides. However, the mechanisms by which Curcuma longa or curcumin exerts cytotoxicity in pests are not well understood. In this study, we investigated the potency of Curcuma longa (curcumin) as a natural pesticide employing Sf9 insect line. Autophagy induction effect of curcumin on Spodoptera frugiperda (Sf9) cells was investigated using various techniques including cell proliferation assay, morphology analysis with inverted phase contrast microscope and Transmission Electron Microscope (TEM) analysis. Autophagy was evaluated using the fluorescent dye monodansylcadaverine (MDC). Cell death measurement was examined using 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) within the concentrations of 5-15µg/mL. Curcumin inhibited the growth of the Sf9 cells and induced autophagic cell death in a time and dose dependent manner. Staining the cells with MDC showed the presence of autophagic vacuoles while increased in a dose and time dependent manner. At the ultrastructural level transmission electron microscopy, cells revealed massive autophagy vacuole accumulation and absence of chromatin condensation. Protein expression levels of ATG8-I and ATG8-II, well-established markers of autophagy related protein were elevated in a time dependent manner after curcumin treatment. The present study proves that curcumin induces autophagic cell death in Sf9 insect cell line and this is the first report of cytotoxic effect of curcumin in insect cells and that will be utilized as natural pesticides in future.

Distribution of neuropeptides in the antennal lobes of male Spodoptera littoralis.

Olfaction is an important sensory modality that regulates a plethora of behavioural expressions in insects. Processing of olfactory information takes place in the primary olfactory centres of the brain, namely the antennal lobes (ALs). Neuropeptides have been shown to be present in the olfactory system of various insect species. In the present study, we analyse the distribution of tachykinin, FMRFamide-related peptides, allatotropin, allatostatin, myoinhibitory peptides and SIFamide in the AL of the male Egyptian cotton leafworm, Spodoptera littoralis. Immunocytochemical analyses revealed that most neuropeptides were expressed in different subpopulations of AL neurons. Their arborisation patterns within the AL suggest a significant role of neuropeptide signalling in the modulation of AL processing. In addition to local interneurons, our analysis also revealed a diversity of extrinsic peptidergic neurons that connected the antennal lobe with other brain centres. Their distributions suggest that extrinsic neurons perform various types of context-related modulation.

Function and central projections of gustatory receptor neurons on the antenna of the noctuid moth Spodoptera littoralis.

Chemosensory information is crucial for most insects to feed and reproduce. Olfactory signals are mainly used at a distance, whereas gustatory stimuli play an important role when insects directly contact chemical substrates. In noctuid moths, although the antennae are the main olfactory organ, they also bear taste sensilla. These taste sensilla detect sugars and hence are involved in appetitive learning but could also play an important role in food evaluation by detecting salts and bitter substances. To investigate this, we measured the responses of individual taste sensilla on the antennae of Spodoptera littoralis to sugars and salts using tip recordings. We also traced the projections of their neuronal axons into the brain. In each sensillum, we found one or two neurons responding to sugars: one NaCl-responsive and one water-sensitive neuron. Responses of these neurons were dose-dependent and similar across different locations on the antenna. Responses were dependent on the sex for sucrose and on both sex and location for glucose and fructose. We did not observe a spatial map for the projections from specific regions of the antennae to the deutocerebrum or the tritocerebrum/suboesophageal ganglion complex. In accordance with physiological recordings, back-fills from individual sensilla revealed up to four axons, in most cases targeting different projection zones.

Bifenthrin induces DNA damage and autophagy in Spodoptera frugiperda (Sf9) insect cells.

Bifenthrin is one of the most commonly used synthetic pyrethroid insecticides. It targets the nervous system of insects, mainly acting on sodium channels in nerve cell membranes. The high use of bifrenthrin may lead to an increase in pest insect resistance. Additionally, there are only a few studies describing its cytotoxic action. A series of bioassays were carried out, and the results showed that bifenthrin has a significant ability to induce DNA damage and the inhibition of viability in Spodoptera frugiperda (Sf9) cells. Monodansylcadaverine staining and transmission electron microscope assays were used to observe significant levels of autophagosomes and mitochondrial dysfunction in the cytoplasm. Additionally, western blot analysis showed an upregulation in LC3-II and beclin-1 protein expression and a downregulation in p62 expression, which contributed to the cytotoxic effect of bifenthrin on Sf9 cells. Overall, bifenthrin significantly impacts the viability of Sf9 cells by inducing DNA damage and autophagy. These results provide a theoretical basis for understanding bifrenthin's mechanism of cytotoxicity.

Pest and natural enemy: how the fat bodies of both the southern armyworm Spodoptera eridania and the predator Ceraeochrysa claveri react to azadirachtin exposure.

The effects of biopesticides on insects can be demonstrated by morphological and ultrastructural tools in ecotoxicological analysis. Azadirachtin-based products are widely used as biopesticides, affecting numerous insect populations. Through morphological biomarkers, this study aimed to characterize the fat bodies of both the southern armyworm Spodoptera eridania and the predator Ceraeochrysa claveri after chronic exposure to azadirachtin. Larvae of S. eridania and C. claveri were fed with fresh purple lettuce leaves (Lactuca sativa) and egg clusters of Diatraea saccharalis treated with azadirachtin solution of 6 mg active ingredient (a.i.)/L and 18 mg a.i./L for 7 days, respectively. The biological data showed a significant reduction in survival and body mass in S. eridania and cytotoxic effects in the parietal and perivisceral fat bodies in both species. Ultrastructural cell damage was observed in the trophocytes of both species such as dilated cisternae of the rough endoplasmic reticulum and swollen mitochondria. Trophocytes of S. eridania and C. claveri of the parietal and perivisceral layers responded to those injuries by different cytoprotective and detoxification means such as an increase in the amount of cytoplasmic granules containing calcium, expression of heat shock protein (HSP)70/HSP90, and development of the smooth endoplasmic reticulum. Despite all the different means of cytoprotection and detoxification, they were not sufficient to recover from all the cellular damages. Azadirachtin exhibited an excellent performance for the control of S. eridania and a moderate selectivity for the predator C. claveri, which presents better biological and cytoprotective responses to chronic exposure to azadirachtin.

Molecular cloning and expression patterns of a putative olfactory diacylglycerol kinase from the noctuid moth Spodoptera littoralis.

In the aim of the characterization of the molecular actors of insect olfactory transduction, we have cloned the full cDNA encoding a Spodoptera littoralis diacylglycerol kinase (DGK) named SlDGK. In male adults, SlDGK transcript was detected predominantly in the brain and in the olfactory sensilla trichodea located on the antennae. SlDGK expression was first detected at day 3 of the pupal stage, then reached a maximum at the end of this stage and was maintained at this level during the adult period. These data provide the first molecular characterization of a DGK potentially involved in the regulation of signalling pathways responsible for the establishment and/or the functioning of the olfactory system in Lepidoptera.

Disruption of Spodoptera exigua larval development by silencing chitin synthase gene A with RNA interference.

RNA interference (RNAi) is a powerful tool for rapidly analyzing gene functions. However, little is known about the possible use of dsRNA/siRNA as a pest control method. Here, we demonstrate that dsRNA/siRNA can induce the silence of chitin synthase gene A (CHSA), which is an important gene for the growth and development of cuticles and trachea in beet armyworm, Spodoptera exigua. Based on the in vitro RNAi experiments in an insect cell line (Trichoplusia ni High 5), in vivo RNAi was performed by injecting synthesized dsRNA/siRNA into the 4th instar larvae of S. exigua. Significantly lower levels of CHSA transcripts were detected. In addition, the cuticle of these insects was disordered and the epithelial walls of larval trachea did not expand uniformly in injected individuals. Moreover, Injections significantly increased abnormalities relative to control larvae. These results highlighted the possibility of dsRNA/siRNA for gene function studies in lepidopteran insects and future pest control.

Toxicity, binding and internalization of the pea-A1b entomotoxin in Sf9 cells.

PA1b (Pea Albumin 1b) is a peptide toxin lethal for certain insects. This paper shows that the cultured insect cells Sf9 are sensitive to the toxin and display a high-affinity binding site for PA1b. Mammalian cells are not sensitive and no binding activity was detected. Signs of apoptosis of the Sf9 cells were observed in response to the toxin. The use of this cellular model also demonstrated that PA1b was internalized in the cells, via the binding site, raising the new question of the role of this toxin within the cell, and of the mechanisms leading to cell death.

Optical brightener M2R destroys the peritrophic membrane of Spodoptera exigua (Lepidoptera: Noctuidae) larvae.

Observations under an environmental scanning electron microscope showed that the peritrophic membrane (PM) of Spodoptera exigua (Hübner) was smooth without pores, but there were many pores and slits in the PM of larvae fed with 10 g L(-1) optical brightener Calcofluor white M2R. After incubation of S. exigua PM in vitro with 10 g L(-1) M2R, a significant amount of proteins was released from the PM structure. M2R disrupted the structure of larval PM, resulting in greatly increased permeability and increased larval susceptibility to Syngrapha falcifera multiple nucleopolyhedrovirus infection. Continuous feeding of larvae on a diet treated with a 10 g L(-1) M2R solution significantly retarded larval development and resulted in high mortality. The destructive effect of M2R on PM was transient and reversible, depending on the presence of M2R within the midgut.

Ultrastructural and developmental toxicity of potato and tomato leaf extracts to beet armyworm, Spodoptera exigua (lepidoptera: noctuidae).

Beet Armyworm, Spodoptera exigua is a herbivorous moth and a serious pest of many economically important plants, which are used as food sources. Because of rigorous standards of food quality, usage of synthetic insecticides in crop protection, against pests, is limited. Solanaceae plant extracts may be a relatively cheap source of efficient natural insecticides that can limit usage of synthetic substances. Their biological activity is not fully known. In particular, ultrastructural studies, using transmission electron microscopy, are not usual. In the present article we describe the effects of sublethal concentrations of tomato and potato leaf extracts against S. exigua. Acute lethal effects were not observed. Both extracts exerted similar effects within midgut and fat body cells. Midgut cells were not significantly altered while fat body cells showed prominent swelling of nuclear envelope and endoplasmic reticulum, vacuolization of mitochondria and fusion of fat droplets. These changes were much more intensive within groups exposed to potato than tomato extracts at highest concentration at least. Light microscopy was used to observe and document developmental alterations of S. exigua exposed to potato and tomato leaf extracts. Potato leaf extracts significantly decreased hatching success and caused morphological malformations of imagoes. Among them, malformations of wings were the most prominent. Interestingly, these effects were not observed within populations exposed to tomato extracts at highest concentration at least.

Linoleic acid peroxidation by Solanum tuberosum lipoxygenase was activated in the presence of human 5-lipoxygenase-activating protein.

The present investigation describes the ability of human 5-lipoxygenase-activating protein (FLAP) to activate a plant 5-lipoxygenase. The presence of an active recombinant human FLAP in the 100000xg membrane fraction of infected Sf9 cells led to a specific increase in 9-hydroperoxyoctadecadienoic acid (9-HPOD) synthesis (+68%) or in 5-hydroperoxyeicosatetraenoic acid (5-HPETE) synthesis (+68%), after action of Solanum tuberosum tuber 5-lipoxygenase (S.t.LOX) on linoleic acid (natural plant lipoxygenase substrate) or on arachidonic acid. On the contrary, the presence of non-transfected membranes obtained from non-infected Sf9 cells led to an inhibition of lipoxygenase activity. MK-886, a potent inhibitor of leukotriene biosynthesis, blocked the FLAP dependent S.t.LOX activation after preincubation with FLAP transfected membranes. In conclusion, this study demonstrates that a recombinant human FLAP can stimulate a lipoxygenase other than mammalian 5-lipoxygenase (S.t.LOX) by using different polyunsaturated fatty acids as substrates.

Stage-dependent strategies of host invasion in the egg-larval parasitoid Chelonus inanitus.

Chelonus inanitus (Braconidae) is a solitary egg-larval parasitoid which lays its eggs into eggs of Spodoptera littoralis (Noctuidae); the parasitoid larva then develops in the haemocoel of the host larva. Host embryonic development lasts approx. 3.5 days while parasitoid embryonic development lasts approx. 16 h. All stages of host eggs can be successfully parasitized, and we show here that either the parasitoid larva or the wasp assures that the larva eventually is located in the host's haemocoel. (1) When freshly laid eggs, up to almost 1-day-old, are parasitized, the parasitoid hatches while still in the yolk and enters the host either after waiting or immediately through the dorsal opening. (2) When 1-2-day-old eggs are parasitized, the host embryo has accomplished final dorsal closure and is covered by an embryonic cuticle when the parasitoid hatches; in this case the parasitoid larva bores with its moving abdominal tip into the host. (3) When 2.5-3.5-day-old eggs are parasitized, the wasp oviposits directly into the haemocoel of the host embryo; from day 2 to 2.5 the embryo is still very small and the wasps, after probing, often restrain from oviposition for a few hours.

Creation of a fully active, cytosolic form of human type I 3beta-hydroxysteroid dehydrogenase/isomerase by the deletion of a membrane-spanning domain.

Human 3beta-hydroxysteroid dehydrogenase/steroid Delta(5)-Delta(4)-isomerase (3beta-HSD/isomerase) is a bifunctional, single enzyme protein that is membrane-bound in the endoplasmic reticulum (microsomes) and mitochondria of cells in the placenta (type I) and in the adrenals and gonads (type II). Two membrane-binding domains (residues 72-89 and 283-310) have been predicted by analyses of hydrophobicity in the type I and II isoenzymes (90% regional homology). These putative membrane domains were deleted in the cDNA by PCR-based mutagenesis, and the two mutant enzymes were expressed by baculovirus in insect Sf9 cells. Differential centrifugation of the Sf9 cell homogenate containing the 283-310 deletion mutant revealed that 94% of the 3beta-HSD and isomerase activities were in the cell cytosol, 6% of the activities were in the microsomes, and no activity was in the mitochondria. This is the opposite of the subcellular distribution of the wild-type enzyme with 94% of the activities in the microsomes and mitochondria and only 6% activity in the cytosol. The organelle distribution of the 72-89 deletion mutant lies between these two extremes with 72% of the enzyme activity in the cytosol and 28% in the microsomes/mitochondria. The integrity of the subcellular organelle preparations was confirmed by electron microscopy. Western immunoblots confirmed the presence of the 283-310 deletion mutant enzyme and the absence of the wild-type enzyme in the insect cell cytosol. The unpurified, cytosolic 383-310 deletion mutant exhibited 3beta-HSD (22 nmol/min per mg) and isomerase (33 nmol/min per mg) specific activities that were comparable with those of the membrane-bound, wild-type enzyme. The isomerase reaction of the cytosolic 283-311 deletion mutant requires activation by NADH just like the isomerase of the microsomal or mitochondrial wild-type enzyme. In contrast, the 72-89 deletion mutant had low 3beta-HSD and isomerase specific activities that were only 12% of the wild-type levels. This innovative study identifies the 283-310 region as the critical membrane domain of 3beta-HSD/isomerase that can be deleted without compromising enzyme function. The shorter 72-89 region is also a membrane domain, but deletion of this NH(2)-terminal region markedly diminishes the enzyme activities. Purification of the active, cytosolic 283-310 deletion mutant will produce a valuable tool for crystallographic studies that may ultimately determine the tertiary/quaternary structure of this key steroidogenic enzyme.

Morphological changes of the insect cells in the baculovirus system as a function of v-myb and c-myb inserts expression and topographic localization of v-Myb and c-Myb proteins.

Structural changes of insect cells Spodoptera frugiperda in the baculovirus expression system after expression of v-myb oncogene and c-myb protooncogene inserts were studied by means of electron microscopy. Expression of v-myb gene insert was accompanied by extensive changes in cell structure, when compared with those of the noninfected and wild-type virus-infected cells. Enormous increase in nuclear content was apparent within 48 hrs after infection, along with changes in nucleolus appearance. Large ring-shaped nucleoli, compact nucleoli and nucleoli with nucleolonemas were detected together with dense nucleolus of normal appearance and small nucleolar structures localized in the nuclear periphery. The cytoplasm practically disappeared 72 hrs after infection. Morphological changes of insect cells expressing the c-myb gene were significantly less distinct, but more frequent unraveling of nucleoli was observed. Both v-Myb and c-Myb proteins were localized in the nucleus of infected cells as was revealed by fluorescence microscopy and electron microscopy. c-Myb marker decorated distinctly the ring-shaped area of nucleolus with some less intensive labelling of the inner part of nucleolus and proximal area on nuclear membrane. v-Myb protein revealed predominantly more compact and homogeneous distribution inside the nucleolus but a small proportion of it was also detected outside the nucleolus in the nuclear compartment. The data obtained on insect cells suggest that Myb proteins may participate also in the processes in which the nucleolus plays a role.

A hybrid baculovirus-bacteriophage T7 transient expression system.

A hybrid recombinant baculovirus-bacteriophage T7 expression system was developed for transient expression in insect cells of plasmids with foreign genes provided with a T7 promoter. The coding sequence for T7 RNA polymerase, with or without a nuclear localization signal, was inserted into the genome of Autographa californica nuclear polyhedrosis virus. Recombinant viruses stably expressed T7 RNA polymerase in insect cells. Upon transfection of infected insect cells with plasmids containing the genes for chloramphenicol acetyltransferase (CAT), the hepatitis B virus precore-, core- or e- antigens under control of the T7 promoter, transient expression of these genes was detected by ELISA. The results obtained indicate that this baculovirus/T7 system provides a simple and widely applicable tool for transient gene expression studies.

The intracellular domain of the rabbit prolactin receptor is able to promote the secretion of a passenger protein via an unusual secretory pathway in lepidopteran cells.

We have previously shown that the intracellular domain of the rabbit prolactin receptor (rbPRL-R), lacking typical signal sequences, was very efficiently secreted into the culture medium when expressed in the baculovirus-insect cell system. We have sought to take advantage of this characteristic for secreting cytoplasmic or nuclear proteins. We have constructed a series of recombinant viruses expressing a foreign gene product fused to the intracellular domain of rbPRL-R. Two passenger genes were used, one encoding a cytoplasmic protein (cyclin B) and the other a nuclear protein (cyclin A). The intracellular domain of rbPRL-R was able to promote the export of these two chimeric proteins with a very high efficiency. This new system should prove useful for secretion of proteins which do not require the post-translational modifications of the classical secretory pathway to be fully active.

Quantitative and ultrastructural changes in the haemocytes of Spodoptera littoralis (Boisd.) treated individually or in combination with Spodoptera littoralis multicapsid nucleopolyhedrovirus (SpliMNPV) and azadirachtin.

The total haemocyte count (THC) and the possible ultrastructural alterations induced in the haemocytes of the fourth larval instars of the Egyptian cotton leafworm, Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae), 96 h post-feeding on a semi-synthetic diet, treated with the LC50 of Spodoptera littoralis multicapsid nucleopolyhedrovirus (SpliMNPV) and the LC50 of azadirachtin alone, and the LC25 of SpliMNPV combined with the LC25 of azadirachtin were studied and compared to the control. Single treatment with the virus and azadirachtin or combined treatment significantly decreased the THC compared to the control. There are five types of haemocytes in S. littoralis: prohaemocytes, plasmatocytes, granulocytes, spherulocytes and oenocytoids. The most common symptoms in granulocytes and plasmatocytes, the main affected cell types, due to viral infection were the presence of virogenic stroma, peripheral dispersion of the chromatin and disappearance of the nucleoli. However, the most common symptoms in these two types of haemocytes due to treatment with azadirachtin were the presence of rough endoplasmic reticulum filled with fibrous materials, due to probably apoptosis, in their cisternae and disorganization of mitochondria (looped, vacuolated and swollen). In addition, the cytoplasm of granulocytes was vacuolated with the appearance of autophagic lysosomes, while plasmatocytes showed ruptured cell membrane and folded nuclear envelope. Combined treatment with the NPV and azadirachtin induced the same pathological changes which were recorded from individual treatment with the virus or azadirachtin to the same haemocytes. It can be concluded that the change in the THC and ultrastructure of granulocytes and plasmatocytes may affect the cellular-mediated immune response in S. littoralis. Moreover, it seems likely that mitochondria were the target site of azadirachtin, as they were affected in both granulocytes and plasmatocytes treated with azadirachtin alone or in combination with SpliMNPV.

Molecular identification and characterization of the Orco orthologue of Spodoptera litura.

A highly conserved and broadly expressed receptor protein Orco (olfactory coreceptor) is crucial for insect olfaction, and an orthologue of Orco has been identified in several insect species. Here we report the identification and characterization of Orco from Spodoptera litura. The protein displays high primary amino acid sequence conservation with other previously identified Orco orthologues. Bioinformatic analysis revealed that it has common features with other members of the Orco subfamily: seven-transmembrane domains with intracellular N-terminus and extracellular C-terminus. The transcript was detected in abundance in the chemosensory organs of the antennae of both male and female adults by real-time polymerase chain reaction analysis, and was localized at the bases of all categories of olfactory sensilla through in situ hybridization.