Translational reprogramming of Zea mays-fed larval midgut of Spodoptera litura indicates non-adaptive response

Structure and expression of glutathione S-transferase genes from the midgut of the Common cutworm, Spodoptera litura (Noctuidae) and their response to xenobiotic compounds and bacteria

Glutathione S-transferase SlGSTE1 in Spodoptera litura may be associated with feeding adaptation of host plants

Copper-induced H2O2 accumulation confers larval tolerance to xanthotoxin by modulating CYP6B50 expression in Spodoptera litura

Functional characterization of CYP6AE subfamily P450s associated with pyrethroid detoxification in Spodoptera litura

This study presents the green synthesis of silver nanoparticles (AgNPs) using Caryota urens leaf extract, emphasizing its role as a reducing and stabilizing agent. The synthesized AgNPs, characterized by UV-vis spectroscopy, FTIR, XRD, and SEM, were predominantly spherical, ranging from 10 to 50 nm. Biochemical assays revealed that bioactive molecules like flavonoids and phenolics contributed significantly to the formation and stability of AgNPs. The bioassay results demonstrated potent larvicidal effects against Spodoptera litura. The mortality rate was dose-dependent, with 95% mortality achieved at 100 mg/mL after 48 h (p < 0.001). The LC50 and LC90 values were 40 and 80 mg/mL, respectively. The nanoparticles also inhibited feeding and egg-laying behaviors in S. litura, reducing oviposition by 82% at the highest concentration. Biochemical studies indicated significant inhibition of acetylcholinesterase (AChE) and elevated glutathione S-transferase (GST) activity, suggesting oxidative stress and disrupted neuromuscular coordination. Histopathological analyses showed severe tissue damage in larval midguts, including epithelial disintegration, mitochondrial swelling, and necrosis. These structural changes were consistent with nanoparticle-induced oxidative damage and enzymatic inhibition.

Role of the epsilon glutathione S-transferases in xanthotoxin tolerance in Spodoptera litura

Susceptibility of the armyworm, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) to nucleopolyhedrovirus was evaluated using droplet feeding methods. S. litura was originally collected from the field in Bantul and has been reared continuously in the laboratory using artificial diet. The tested instars were exposed a series concentration of nucleopolyhedrovirus (2×103, 2×104, 2×105, 2×106, 2×107, 2×108, 2×109 PIB/ml) which were added with Tinopal 0,5% and 1%. The result indicated that the larval mortality of 3rd, 4th, and 5th instars Tinopal, significantly different with the addition of 1% Tinopal. This addition increased the effectiveness of NPV for 235, 25117, and 6.6 million fold. The observation of the larval midgut which was treated by Tinopal, showed that Tinopal physically disrupt the peritrophic membrane. Therefore, it can be suggested that the Tinopal facilitates the entry of NPV to the host insect.

By using shotgun HPLC-ESI-MS proteomics approach, 2043 peptides were identified from the midgut of Spodoptera litura larvae at the sixth instar feeding stage, out of which 1489 (72.9%) were found to have their homologues in the public protein databases and 842 had identities of molecular functions. Seven-hundred forty-one peptides were annotated according to Gene Ontology Annotation in terms of molecular function, biological process, and cellular localization, with 336 and 251 peptides being related to catalytic activity and binding activity, respectively. Most of the catalytic proteins had activity of hydrolases, oxidoreductases and transferases and most of the binding proteins were involved in protein-binding activity. Among the annotated peptides, 487 were classified into different cellular processes and 490 were classified to locate in the cytoplasm. Nonredundant enzymes associated with the metabolisms of carbohydrates, lipids and fatty acids, amino acids and proteins, translation, transport, and stress resistance were identified. Presence and expression at high levels of numerous enzymes of glycolysis pathway, synthesis of proteins, and absorption and transport of fatty acids and lipids indicate that active metabolism processes of carbohydrates, proteins, and lipids occurred in the midgut of sixth instar feeding larvae of S. litura. The protein profile provides a basis for further study of the physiological events in the midgut of S. litura.

BackgroundSterol carrier protein-2/3-oxoacyl-CoA thiolase (SCPx) gene has been suggested to be involved in absorption and transport of cholesterol. Cholesterol is a membrane component and is a precursor of ecdysteroids, but cannot be synthesized de novo in insects. However, a direct association between SCPx gene expression, cholesterol absorption and development in lepidopteran insects remains to be experimentally demonstrated.ResultsAn SCPx cDNA (SlSCPx) cloned from the common cutworm, Spodoptera litura, was characterized. The SlSCPx cDNA encoded a 535-amino acid protein consisting of a 3-oxoacyl-CoA thiolase (SCPx-t) domain and a SCP-2 (SCPx-2) domain. SlSCPx mRNA was expressed predominately in the midgut, while SlSCPx-2 mRNA was detected in the midgut, fat body and epidermis and no SlSCPx-t mRNA was detected. A 58-kDa full-length SCPx protein and a 44-kDa SCPx-t protein were detected in the midgut of sixth instar larvae when the anti-SlSCPx-t antibody was used in western blotting analysis; a 16-kDa SCP-2 protein was detected when anti-SlSCPx-2 antibody was used. SlSCPx protein was post-translationally cleaved into two smaller proteins, SCPx-t and SCPx-2. The gene appeared to be expressed into two forms of mRNA transcripts, which were translated into the two proteins, respectively. SlSCPx-t and SlSCPx-2 proteins have distinct and different locations in the midgut of sixth instar larvae. SlSCPx and SlSCPx-t proteins were detected predominately in the cytoplasm, whereas SlSCPx-2 protein was detected in the cytoplasm and nuclei in the Spli-221 cells. Over-expression of SlSCPx and SlSCPx-2 proteins enhanced cholesterol uptake into the Spli-221 cells. Knocking-down SlSCPx transcripts by dsRNA interference resulted in a decrease in cholesterol level in the hemolymph and delayed the larval to pupal transition.ConclusionSpatial and temporal expression pattern of this SlSCPx gene during the larval developmental stages of S. litura showed its specific association with the midgut at the feeding stage. Over-expression of this gene increased cholesterol uptake and interference of its transcript decreased cholesterol uptake and delayed the larval to pupal metamorphosis. All of these results taken together suggest that this midgut-specific SlSCPx gene is important for cholesterol uptake and normal development in S. litura.

Juvenile hormone (JH) is one of the key insect hormones that regulate metamorphosis. Juvenile hormone diol kinase (JHDK) is an enzyme involved in JH metabolism and catalyzes JH diol to form a polar end product, JH diol phosphate that has no JH activity. In this study, a JHDK complementary DNA (cDNA) was cloned from Spodoptera litura and the structure and expression of the gene was characterized. The cDNA was 714 base pairs in length and encoded a protein of 183 amino acids with a molecular mass of 21 kDa and an isoelectric point of 4.55. Based on the structure, three putative calcium binding motifs and guanosine triphosphate-binding motifs were predicted in the protein. Modeling of the 3-D structure showed that the protein consisted of eight α-helixes linked with loops, with no β-sheets. The gene was expressed in the epidermis, fat body and midgut of fifth and sixth instar larvae. The expression level in the epidermis was lower than in the fat body and midgut. The gene was expressed at higher levels at the early stages than in the later stages of fifth and sixth instar midgut and fat body. The results suggest that this gene may be involved in the regulation of the JH titer in larvae of S. litura.

Herbivore-induced tomato plant volatiles lead to the reduction of insecticides susceptibility in Spodoptera litura

Serine proteases play important roles in food digestion and immune response during insect development. A full-length cDNA (Slctlp1) encoding a chymotrypsin-like serine protease was cloned from Spodoptera litura and characterized for its cDNA structure, developmental and induced expression and localization. The deduced protein of the Slctlp1 cDNA contains a catalytic triad and a substrate specificity pocket found in most of the serine proteases. Both the transcripts and protein of Slctlp1 were predominately expressed in the midgut at the feeding stages during the larval development. Immunohistochemistry analysis indicated that the SLCTLP1 protein was predominately present in the midgut of the 6th instar feeding larvae. Starvation suppressed the expression of Slctlp1 gene and protein in 6th instar larvae and the protein expression was increased again by re-feeding the insect. The results suggest that the cloned Slctlp1 cDNA may be involved in food protein digestion at the feeding stages during larval development.

Insect cytochrome P450 monooxygenases (CYPs or P450s) play an important role in detoxifying insecticides leading to resistance in insect populations. A polyphagous pest, Spodoptera litura, has developed resistance to a wide range of insecticides. In the present study, a novel P450 gene, CYP321B1, was cloned from S. litura. The function of CYP321B1 was assessed using RNA interference (RNAi) and monitoring resistance levels for three commonly used insecticides, including chlorpyrifos, β-cypermethrin and methomyl. The full-length complementary DNA sequence of CYP321B1 is 1814 bp long with an open reading frame of 1488 bp encoding 495 amino acid residues. Quantitative reverse-transcriptase polymerase chain reaction analyses during larval and pupal development indicated that CYP321B1 expression was highest in the midgut of fifth-instar larvae, followed by fat body and cuticle. The expression of CYP321B1 in the midgut was up-regulated by chlorpyrifos, β-cypermethrin and methomyl with both lethal concentration at 15% (LC15 ) (50, 100 and 150 μg/mL, respectively) and 50%(LC50 ) dosages (100, 200 and 300 μg/mL, respectively). Addition of piperonyl butoxide (PBO) significantly increased the toxicity of chlorpyrifos, β-cypermethrin and methomyl to S. litura, suggesting a marked synergism of the three insecticides with PBO and P450-mediated detoxification. RNAi-mediated silencing of CYP321B1 further increased mortality by 25.6% and 38.9% when the fifth-instar larvae were exposed to chlorpyrifos and β-cypermethrin, respectively, at the LC50 dose levels. The results demonstrate that CYP321B1 might play an important role in chlorpyrifos and β-cypermethrin detoxification in S. litura.

A chymotrypsin-like serine protease cDNA involved in food protein digestion in the common cutworm, Spodoptera litura: Cloning, characterization, developmental and induced expression patterns, and localization

Standard leaf dip bioassay experiments were conducted to study the insecticide resistance in Spodoptera litura (F.) in soybean. Chlorantraniliprole was the most toxic while, cypermethrin and organophosphates (profenophos and triazophos) recorded least toxicity against S. litura populations. Field population of S. litura showed the highest resistance to cypermethrin with resistance ratios (RRs) ranging from 244 to 376. The field populations collected from three locations of Southern Rajasthan exhibited low resistance to newer insecticides, chlorantraniliprole and spinetoram with Lab-SS strain. All the enzyme activities were significantly higher in the whole larval extracts and midgut extracts of S. litura larvae when compared to the Lab-SS strain. Analysis of the carboxyl esterase enzyme in the native PAGE assay revealed that the electrophoretic profiles showed presence of different band regions (EST 1 to EST 7) with esterase activity in the field populations along with the laboratory susceptible strain. The susceptibility of three field populations to different group of insecticides along with the profile of detoxification enzymes indicate the need to formulate a region-specific insecticide resistance management (IRM).