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Sensory genes identification with head transcriptome of the migratory armyworm, Mythimna separata.

ABSTRACT: Sensory system plays important roles in a wide array of insect's behavior and physiological events, including the host landing and locating, feeding, flying, sex responding, mating and oviposition which happen independently and in sequence. The armyworm Mythimna separata (Lepidoptera: Noctuidae) of migratory insect is destructive for alimentarn crop and economic crop throughout the world. Here we present the high throughput sequencing of the head transcriptome and identify members of the major sensory genes which are crucial for armyworm's success worldwide, including 8 opsins, 22 chemosensory proteins, 50 odorant binding proteins, 60 odorant receptors, 8 gustatory receptors, 24 ionotropic receptors, and 2 sensory neuron membrane proteins. It is worth noting that a duplication of the LW opsin gene exists in this insect. Several genes were clustered with functionally validated genes, such as Co-receptors of OR and IR, PBPs, PRs, CO2 GRs, bitter GRs and sweet GRs, were also identified. The transcriptome gene library provided the basis for further studies that elucidate the fundamental molecular mechanism of biology and control in M. separata. Our research exhibits the first comprehensive catalogue of the sensory genes fundamental for success and distribution in M. separata, which are potential novel targets for pest control strategies.

SUBMITTER: Liu Z

PROVIDER: S-EPMC5384095 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Identification and characteristics of a novel cecropin from the armyworm, Mythimna separata.

Project description:BackgroundThe recent emergence of antibiotic-resistant strains of bacteria has increased the need to develop effective alternatives to antibiotics. Antimicrobial peptides have been considered as a promising product with several advantages.ResultsIn this present study, we identified a novel cecropin from the armyworm, Mythimna separata (armyworm cecropin 1, AC-1) by transcriptome sequencing and multi-sequence alignment analysis. The AC-1 precursor comprised 63 amino acid residues, containing a conserved cleavage site of the signal peptide, Ala23-Pro24, while the mature AC-1 included 39 amino acid residues. Chemically synthesized AC-1 exhibited low hemolytic activity against chicken red blood cells, low cytotoxicity against swine testis cells, and effective antimicrobial activity against Salmonella, Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa. Its antimicrobial activity against Salmonella remained after incubation for 1 h at 100 °C or in 250 mM NaCl, KCl, or MgCl2 solution, implying good thermal- and salt-resistant stabilities. The bactericidal effect of AC-1 on E. coli gradually increased with increasing AC-1 concentration, resulting in deformation, severe edema, cytolysis, cell membrane damage, and reducing intracellular electron density. Additionally, recombinant AC-1 protein expressed in E. coli was digested by enterokinase protease to obtain AC-1, which showed similar antimicrobial activity against E. coli to chemically synthesized AC-1.ConclusionsThis study identified a novel antimicrobial peptide that may represent a potential alternative to antibiotics.

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De Novo Assembly of the Transcriptome for Oriental Armyworm Mythimna separata (Lepidoptera: Noctuidae) and Analysis on Insecticide Resistance-Related Genes.

Project description:Mythimna separata walker (Lepidoptera: Noctuidae) is a polyphagous pest of nearly 100 families of more than 300 kinds of food and industrial crops. So far, both nucleotide and protein sequence information has been rarely available in database for M. separata, strictly limiting molecular biology research in this insect species. In this study, we carried out a transcriptome sequencing for M. separata The sequencing and subsequent bioinformatics analysis yielded 69,238 unigenes, among which 45,227 unigenes were annotated to corresponding functions by blasting with high homologous genes in database, giving annotation rate of 65.32%. Several lepidopteran insects gave best matches with the transcriptome data. To gain insight into the mechanism of insecticide resistance in M. separata, 15 families of genes encoding insecticide resistance-related proteins were investigated. Substantial numbers of unigenes in these families were identified in the transcriptome data, and 17 out of 21 selected unigenes were successfully amplified. Expressions of most of these genes were detected at larval stages and in gut tissue, as was consistent with their putative involvement in insecticide resistance. Our study provides most comprehensive transcriptome data for M. separata to date, and also provides reference sequence information for other Noctuidae family insects.

| S-EPMC5026479 | biostudies-literature

Characterization of the Adult Head Transcriptome and Identification of Migration and Olfaction Genes in the Oriental Armyworm Mythimna separate.

Project description:The oriental armyworm Mythimna separate is an economically important insect with a wide distribution and strong migratory activity. However, knowledge about the molecular mechanisms regulating the physiological and behavioural responses of the oriental armyworm is scarce. In the present study, we took a transcriptomic approach to characterize the gene network in the adult head of M. separate. The sequencing and de novo assembly yielded 63,499 transcripts, which were further assembled into 46,459 unigenes with an N50 of 1,153?bp. In the head transcriptome data, unigenes involved in the 'signal transduction mechanism' are the most abundant. In total, 937 signal transduction unigenes were assigned to 22 signalling pathways. The circadian clock, melanin synthesis, and non-receptor protein of olfactory gene families were then identified, and phylogenetic analyses were performed with these M. separate genes, the model insect Bombyx mori and other insects. Furthermore, 1,372 simple sequence repeats of 2-6?bp in unit length were identified. The transcriptome data represent a comprehensive molecular resource for the adult head of M. separate, and these identified genes can be valid targets for further gene function research to address the molecular mechanisms regulating the migratory and olfaction genes of the oriental armyworm.

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Identification of 35 C-Type Lectins in the Oriental Armyworm, Mythimna separata (Walker).

Project description:Insect C-type lectins (CTLs) play vital roles in modulating humoral and cellular immune responses. The oriental armyworm, Mythimna separata (Walker) (Lepidoptera: Noctuidae) is a migratory pest that causes significant economic loss in agriculture. CTLs have not yet been systematically identified in M. separata. In this study, we first constructed a transcriptome of M. separata larvae, generating a total of 45,888 unigenes with an average length of 910 bp. Unigenes were functionally annotated in six databases: NR, GO, KEGG, Pfam, eggNOG, and Swiss-Prot. Unigenes were enriched in functional pathways, such as those of signal transduction, endocrine system, cellular community, and immune system. Thirty-five unigenes encoding C-type lectins were identified, including CTL-S1~CTL-S6 (single CRD) and IML-1~IML-29 (dual CRD). Phylogenetic analyses showed dramatic lineage-specific expansions of IMLs. Sequence alignment and structural modeling identified potential ligand-interacting residues. Real-time qPCR revealed that CTL-Ss mainly express in eggs and early stage larvae, while IMLs mainly express in mid-late-stage larvae, pupae, and adults. In naïve larvae, hemocytes, fat body, and epidermis are the major tissues that express CTLs. In larvae challenged by Escherichia coli, Staphylococcus aureus, or Beauveria bassiana, the expression of different CTLs was stimulated in hemocytes, fat body and midgut. The present study will help further explore functions of M. separata CTLs.

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Screening potential reference genes for quantitative real-time PCR analysis in the oriental armyworm, Mythimna separata.

Project description:The oriental armyworm, Mythimna separata, is a major insect pest in China and other Asian countries. Unfortunately, suitable reference genes for quantitative real-time PCR (qRT-PCR) have not been previously identified in M. separata for normalizing target gene expression. In this study, we evaluated the expression stability of eight candidate genes (18S, ACT, EF1-α, GAPDH, RPS7, RPS13, RPL32 and TUB) in M. separata using the comparative ΔCt method, BestKeeper, Normfinder geNorm and ReFinder, a comprehensive software platform. The results indicated that the appropriate reference gene varied depending on the experimental conditions. We found that ACTIN, EF1-α and TUB were optimal for different developmental stages; TUB, RPS13 and EF1-α showed the most stable expresssion in different tissues; RPS13 and 18S were the best reference genes for monitoring expression under high temperature conditions; TUB, RPS13 and RPS7 exhibited the most stable expression under larval-crowding conditions; RPS7, EF1-α, RPL32 and GAPDH were the best for pesticide exposure experiments. This study provides tools for reliable normalization of qRT-PCR data and forms a foundation for functional studies of target gene expression in M. separata.

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A chemosensory protein MsepCSP5 involved in chemoreception of oriental armyworm Mythimna separata.

Project description:Chemosensory proteins (CSPs) have been suggested to perform several functions in insects, including chemoreception. To find out whether MsepCSP5 identified from Mythimna separata shows potential physiological functions in olfaction, gene expression profiles, ligand-binding experiments, molecular docking, RNA interference, and behavioral test were performed. Results showed that MsepCSP5 was highly expressed in female antennae. MsepCSP5 showed high binding affinities to a wide range of host-related semiochemicals, and displayed that 26 out of 35 candidate volatiles were highly bound (Ki < 10 µM) at pH 5.0 rather than pH 7.4. The binding sites of MsepCSP5 to candidate volatiles were well predicted by three-dimensional structure modeling and molecular docking experiments. Pursuing further, biological activities of M. separata to highly bound compounds elicited strong behavioral responses, such as alcoholic compounds displayed strong attractiveness whereas terpenes showed repellency to M. separata. The transcript expression level of MsepCSP5 gene significantly decreased after injecting target dsRNAs, and resulted in non-significant preference responses of M. separata to semiochemicals, such as 3-pentanol and 1-octene-3-ol. In conclusion, MsepCSP5 may involve in semiochemical reception of M. separata.

| S-EPMC6299372 | biostudies-literature

Identification of host plant use of adults of a long-distance migratory insect, Mythimna separata.

Project description:Adults of many insect species often become contaminated with pollen grains when feeding. Identification of plant hosts for M. separata moths could increase our understanding of their geographic origin and the coevolution of M. separata moths and their host plants. However, identifying the diet of noctuid moths using traditional direct observation is limited by their nocturnal and flight habits. In this study, we used core barcode markers and pollen morphology to identify pollen species. We found pollen from 13 plant species belonging to nine families on trapped M. separata moths, mainly from Angiosperm, Dicotyledoneae. Pollen was found on 14.4% and 12.3% of females and males, respectively, and the amount of pollen transported varied with the body part, with the most pollen on the proboscis. We were able to determine from this that the moths visited woody plants more than herbaceous plants, but not significantly so, and that they carried more pollen earlier in the migration season. In this study, we clarified the species and frequencies of pollen deposition on M. separata moths. These findings improve our understanding of the coevolution of the moths and their host plants. Identification of plant hosts for adult moths provides a new means of studying noctuid moth-host plant interactions, and informs the development of more efficient management practices for M. separata.

| S-EPMC5584948 | biostudies-literature

Identification of candidate chemosensory genes in Mythimna separata by transcriptomic analysis.

Project description:BackgroundThe oriental armyworm, Mythimna separata, is an economically important and common Lepidopteran pest of cereal crops. Chemoreception plays a key role in insect life, such as foraging, oviposition site selection, and mating partners. To better understand the chemosensory mechanisms in M. separata, transcriptomic analysis of antennae, labial palps, and proboscises were conducted using next-generation sequencing technology to identify members of the major chemosensory related genes.ResultsIn this study, 62 putative odorant receptors (OR), 20 ionotropic receptors (IR), 16 gustatory receptors (GR), 38 odorant binding proteins (OBP), 26 chemosensory proteins (CSP), and 2 sensory neuron membrane proteins (SNMP) were identified in M. separata by bioinformatics analysis. Phylogenetic analysis of these candidate proteins was performed. Differentially expressed genes (DEGs) analysis was used to determine the expressions of all candidate chemosensory genes and then the expression profiles of the three families of receptor genes were confirmed by real-time quantitative RT-PCR (qPCR).ConclusionsThe important genes for chemoreception have now been identified in M. separata. This study will provide valuable information for further functional studies of chemoreception mechanisms in this important agricultural pest.

| S-EPMC6030794 | biostudies-literature

Reference Genome Sequences of the Oriental Armyworm, Mythimna separata (Lepidoptera: Noctuidae).

Project description:Lepidopteran insects are an important group of animals, including those used as biochemical and physiological model species in the insect and silk industries as well as others that are major agricultural pests. Therefore, the genome sequences of several lepidopteran insects have been reported. The oriental armyworm, Mythimna separata, is an agricultural pest commonly used to study insect immune reactions and interactions with parasitoid wasps as hosts. To improve our understanding of these research topics, reference genome sequences were constructed in the present study. Using long-read and short-read sequence data, de novo assembly and polishing were performed and haplotigs were purged. Subsequently, gene predictions and functional annotations were performed. To search for orthologs of the Toll and Immune Deficiency (IMD) pathways and for C-type lectins, annotation data analysis, BLASTp, and Hummer scans were performed. The M. separata genome is 682 Mbp; its contig N50 was 2.7 Mbp, with 21,970 genes and 24,452 coding sites predicted. All orthologs of the core components of the Toll and IMD pathways and 105 C-type lectins were identified. These results suggest that the genome data were of sufficient quality for use as reference genome data and could contribute to promoting M. separata and lepidopteran research at the molecular and genome levels.

| S-EPMC9853324 | biostudies-literature

Functional Analysis of MsepOR13 in the Oriental Armyworm Mythimna separata (Walker).

Project description:Olfaction in insects has a critical role in recognizing the host, finding food, and choosing mating partners, as well as avoiding predators. Odorant receptors (ORs), which are housed in the dendritic membrane of sensory neurons and extended into the lymph of sensilla on insect antennae, are participating in the detection of volatile compounds in insects. In the present study, we identified an OR gene, named MsepOR13, in the oriental armyworm Mythimna separata (Walker). Quantitative real-time polymerase chain reaction revealed that MsepOR13 was expressed mainly in the antennae of male and female moths. In in vitro heterologous expression experiments, MsepOR13 was widely tuned to 32 of the 67 different compounds tested. Furthermore, MsepOR13 responded to eugenol at a low concentration of 10-9 M, with an EC50 value of 3.91 × 10-6 M. The high sensitivity suggests an important role for the OR13 gene in the moth olfactory system.

| S-EPMC6465334 | biostudies-literature

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