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Characterization of the complete mitochondrial genome of Asia Corn Borer, Ostrinia furnacalis (Lepidoptera: Crambidae).

ABSTRACT: In this study, the complete mitogenome sequence of Ostrinia furnacalis was described. The assembled mitogenome is 15,241 bp in length with an extreme bias of high AT content (80.9%) (GenBank accession no. MN747041). The mitochondrial genome contains 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (12S rRNA and 16S rRNA) genes, and a control region (D-loop region). The mitochondrial gene order was identical to that observed in most lepidopteran genomes, nine PCGs were located on the H-strand, others were located on the L-strand. 12 PCGs were initiated by typical ATN codons, except for COI with CGA instead. 21 tRNAs had the typical cloverleaf structure, while the DHU arm of the trnS1 gene did not form a stable stem-loop structure. The 'ATAGT(A)'-like motif and a 19 bp poly-T stretch at the down-stream of the rrnS gene were observed in the A + T-rich region. The phylogenetic analysis showed that the relationship of O. furnacalis is very close to the three species in the subfamily Pyraustinae: O. nubilalis, O. penitalis and Loxostege sticticalis, and all the subfamilies of Spilomelinae, Pyraustinae, Crambinae and Nymphulinae within Crambidae formed monophyletic groups with the highest bootstrap value support.

SUBMITTER: Li C

PROVIDER: S-EPMC7748848 | biostudies-literature |

REPOSITORIES: biostudies-literature

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RNA Interference to Control Asian Corn Borer Using dsRNA from a Novel Glutathione-S-Transferase Gene of Ostrinia furnacalis (Lepidoptera: Crambidae).

Project description:Glutathione-S-transferases (GST) comprise a multifunctional protein superfamily, which plays important roles as detoxifiers and antioxidants in insects. The GST in Asian corn borer has not been previously characterized. In this study, we cloned, characterized, and expressed the complete GST genes from the midgut of Asian corn borer. Furthermore, we designed htL4440-OfGST vector to exploit this gene for RNA interference (RNAi) strategy to control this pest. A complete GST cDNA sequence in Asian corn borer was obtained by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends technology. The gene was 887bp in length and contained a 705bp open reading frame and 5' UTR and 3' UTR lengths of 89 and 93bp, respectively. The putative sequence encoded a putative 234 amino acid residue peptide and had a predicted molecular weight of ~26kDa. The GST protein of Asian corn borer is hydrophilic and may have a 30 amino acid signal peptide with a cleavage site between L30 and K31. A recombination vector pET28a-OfGST was constructed for purification and antibody preparation. Western blotting analysis showed that this protein reached the maximum expression level around 24 h in Asian corn borer larvae fed the plant toxin 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one. A second vector, htL4440-OfGST, was constructed to generate the dsRNA of the GST gene. A larval feeding bioassay showed that the expressed dsRNA significantly reduced the detoxification ability of Asian corn borer larvae and increased mortality rate up to 54%. Our data indicated that GST plays very important roles in detoxifying in Asian corn borer and can be used as an RNAi method to control this pest in the field.

| S-EPMC6195416 | biostudies-literature

Male- and Female-Biased Gene Expression of Olfactory-Related Genes in the Antennae of Asian Corn Borer, Ostrinia furnacalis (Guenee) (Lepidoptera: Crambidae).

Project description:The Asian corn borer (ACB), Ostrinia furnacalis (Guenée), is a destructive pest insect of cultivated corn crops, for which antennal-expressed receptors are important to detect olfactory cues for mate attraction and oviposition. Few olfactory related genes were reported in ACB, so we sequenced and characterized the transcriptome of male and female O. furnacalis antennae. Non-normalized male and female O. furnacalis antennal cDNA libraries were sequenced on the Illumina HiSeq 2000 and assembled into a reference transcriptome. Functional gene annotations identified putative olfactory-related genes; 56 odorant receptors (ORs), 23 odorant binding proteins (OBPs), and 10 CSPs. RNA-seq estimates of gene expression respectively showed up- and down-regulation of 79 and 30 genes in female compared to male antennae, which included up-regulation of 8 ORs and 1 PBP gene in male antennae as well as 3 ORs in female antennae. Quantitative real-time RT-PCR analyses validated strong male antennal-biased expression of OfurOR3, 4, 6, 7, 8, 11, 12, 13 and 14 transcripts, whereas OfurOR17 and 18 were specially expressed in female antennae. Sex-biases gene expression described here provides important insight in gene functionalization, and provides candidate genes putatively involved in environmental perception, host plant attraction, and mate recognition.

| S-EPMC4463852 | biostudies-literature

Down-regulation of aminopeptidase N and ABC transporter subfamily G transcripts in Cry1Ab and Cry1Ac resistant Asian corn borer, <i>Ostrinia furnacalis</i> (Lepidoptera: Crambidae).

Project description:The Asian corn borer (ACB), Ostrinia furnacalis (Lepidoptera: Crambidae), is a highly destructive pest of cultivated maize throughout East Asia. Bacillus thuringiensis (Bt) crystalline protein (Cry) toxins cause mortality by a mechanism involving pore formation or signal transduction following toxin binding to receptors along the midgut lumen of susceptible insects, but this mechanism and mutations therein that lead to resistance are not fully understood. In the current study, quantitative comparisons were made among midgut expressed transcripts from O. furnacalis susceptible (ACB-BtS) and laboratory selected strains resistant to Cry1Ab (ACB-AbR) and Cry1Ac toxins (ACB-AcR) when feeding on non-Bt diet. From a combined de novo transcriptome assembly of 83,370 transcripts, ORFs of ≥ 100 amino acids were predicted and annotated for 28,940 unique isoforms derived from 12,288 transcripts. Transcriptome-wide expression estimated from RNA-seq read depths predicted significant down-regulation of transcripts for previously known Bt resistance genes, aminopeptidase N1 (apn1) and apn3, as well as a putative ATP binding cassette transporter group G (abcg) gene in both ACB-AbR and -AcR (log2[fold-change] ≥ 1.36; P < 0.0001). The transcripts that were most highly differentially regulated in both ACB-AbR and -AcR compared to ACB-BtS (log2[fold-change] ≥ 2.0; P < 0.0001) included up- and down-regulation of serine proteases, storage proteins and cytochrome P450 monooxygenases, as well as up-regulation of genes with predicted transport function. This study predicted the significant down-regulation of transcripts for previously known Bt resistance genes, aminopeptidase N1 (apn1) and apn3, as well as abccg gene in both ACB-AbR and -AcR. These data are important for the understanding of systemic differences between Bt resistant and susceptible genotypes.

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Identification of candidate odorant receptors in Asian corn borer Ostrinia furnacalis.

Project description:In lepidopteran insects, odorant receptors are involved in the perception of sex pheromones and general odorants. In the Asian corn borer, Ostrinia furnacalis, although several pheromone receptors have been identified, no general odorant receptor has been reported. In this study, an RNA sequencing analysis was carried out to identify the whole repertoire of the odorant receptors expressed in the antennae of O. furnacalis. Among 12 million reads obtained from the antennae of male and female moths, 52 candidate odorant receptors were identified, including 45 novel ones. Expression levels of candidate odorant receptors were estimated by read mapping and quantitative reverse transcription PCR. These analyses confirmed that the expression of the previously identified pheromone receptors was highly male biased. In contrast, none of the newly identified odorant receptors showed male-biased expression. Three of the newly identified odorant receptors showed female-biased expression. Two of them were the most highly expressed odorant receptors in the female antennae, suggesting that they may be involved in the detection of odorants important for the induction of female-specific behaviors such as oviposition site selection. In addition, candidate genes of 21 ionotropic receptors, 5 gustatory receptors, 2 sensory neuron membrane proteins, and 26 odorant degrading enzymes were identified. Our results provide a basis for further analysis of the chemosensory system in the Ostrinia species.

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Cloning, Expression, and Characterization of Prophenoloxidases from Asian Corn Borer, <i>Ostrinia furnacalis</i> (Gunee).

Project description:Insect phenoloxidase (PO) belongs to the type 3 copper protein family and possesses oxidoreductase activities. PO is typically synthesized as a zymogen called prophenoloxidase (PPO) and requires the proteolytic activation to function. We here cloned full-length cDNA for 3 previously unidentified PPOs, which we named OfPPO1a, OfPPO1b, and OfPPO3, from Asian corn borer, Ostrinia furnacalis (Gunée), in addition to the previously known OfPPO2. These conceptual PPOs and OfPPO2 all contain two common copper-binding regions, two potential proteolytic activation sites, a plausible thiol-ester site, and a conserved C-terminal region but lack a secretion signal peptide sequence at the N-terminus. O. furnacalis PPOs were highly similar to other insect PPOs (42% to 79% identity) and clustered well with other lepidopteran PPOs. RT-PCR assay showed the transcripts of the 4 OfPPOs were all detected at the highest level in hemocytes and at the increased amounts after exposure to infection by bacteria and fungi. Additionally, we established an Escherichia coli (E. coli) expression system to produce recombinant O. furnacalis PPO proteins for future use in investigating their functions. These insights could provide valuable information for better understanding the activation and functioning mechanisms of O. furnacalis PPOs.

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Synergistic Effect of Beauveria bassiana and Trichoderma asperellum to Induce Maize (Zea mays L.) Defense against the Asian Corn Borer, Ostrinia furnacalis (Lepidoptera, Crambidae) and Larval Immune Response.

Project description:Ostrinia furnacalis, is the major pest of maize causing significant yield losses. So far, many approaches have been used to increase the virulence of entomopathogenic fungal isolates. The current study is an attempt to estimate synergistic effect of Beauveria bassiana and Trichoderma asperellum in order to explore larval immune response through RNA sequencing and differentially expression analysis. In vivo synergism was examined in seven proportions (B. bassiana: T. asperellum = 1:1, 1:2, 1:3, 1:4, 4:1, 3:1, 2:1) and in the in vitro case, two inoculation methods were applied: seed coating and soil drenching. Results revealed significant decrease in plant damage and high larval mortality in fungal treatments. Fungal isolates mediated the plant defense by increasing proline, superoxide dismutase (SOD), peroxidase (POD), polyphenol oxidase (PPO) and protease activities. Seed coating method was proved to be the most effective in case of maize endophytic colonization. In total, 59 immune-related differentially expressed genes DEGs were identified including, cytochrome P450, heat shock protein, ABC transporter, cadherin, peptidoglycan recognition protein (PGRP), cuticlular protein, etc. Further, transcriptomic response was confirmed by qRT-PCR. Our results concluded that, coculture of B. bassiana and T. asperellum has the synergistic potential to suppress the immune response of O. furnacalis and can be used as sustainable approach to induce plant resistance through activation of defense-related enzymes.

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Functional Studies of Sex Pheromone Receptors in Asian Corn Borer Ostrinia furnacalis.

Project description:Lepidopteran insects use sex pheromones for sexual communication. Pheromone receptors expressed on peripheral olfactory receptor neurons (ORNs) are critical part to detect the sex pheromones. In genus Ostrinia, several pheromone receptors were functional analyzed in O. nubilalis and O. scapulalis but the knowledge in O. furnacalis was rare. In this study, seven pheromone receptors were deorphanized by heterologous expression system of Xenopus oocytes. Functional types of sensilla trichoidea were classified by single sensillum recordings to interpret the response pattern of olfactory sensory neurons to Ostrinia pheromone components. OfurOR4 and OfurOR6 responded to the major sex pheromone Z/E12-14:OAc. OfurOR4 is the main receptor for both Z/E12-14:OAc and OfurOR6 mainly responded to E12-14:OAc. Functional differentiation of gene duplication were found between OfurOR5a and OfurOR5b. OfurOR5b showed a broad response to most of the pheromone components in O. furnacalis, whereas OfurOR5a was found without ligands. OfurOR7 showed a specific response to Z9-14:OAc and OfurOR8 mainly responded to Z11-14:OAc and E11-14:OAc. OfurOR3 did not respond to any pheromone components. Our results improved the current knowledge of pheromone reception in Ostrinia species which may contribute to speciation.

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Analyses of Cry1Ab binding in resistant and susceptible strains of the European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae).

Project description:Cry1Ab toxin binding analysis was performed to determine whether resistance in laboratory-selected Ostrinia nubilalis strains is associated with target site alteration. Brush border membrane vesicles were prepared using dissected midguts from late instars of susceptible and resistant strains (Europe-R and RSTT) of O. nubilalis. Immunoblot analysis indicated that three different proteins bound to Cry1Ab toxin and were recognized by an anticadherin serum. In a comparison of resistant and susceptible strains, reduced Cry1Ab binding was apparent for all three bands corresponding to cadherin-like proteins in the Europe-R strain, while reduced binding was apparent in only one band for the RSTT strain. Real-time analysis of Cry1Ab binding to gut receptors using surface plasmon resonance suggested slight differences in affinity in both resistant strains. Additional binding analysis was conducted using 125I-labeled Cry1Ab, Cry1Ac, and Cry1Aa. Slight differences were again observed between the resistant and susceptible strains for Cry1Ab binding. However, when binding of 125I-labeled Cry1Aa was tested, a 10-fold reduction in the concentration of binding sites was observed in the Europe-R strain. Expression of the O. nubilalis cadherin gene was similar in both the resistant and susceptible strains and did not account for differences in binding. In combination, the results of the present work suggest that differences in susceptibility to Cry1A toxins in the Europe-R strain of O. nubilalis are associated with altered receptor binding, although the precise nature of this mechanism is still uncertain.

| S-EPMC1538724 | biostudies-literature

Molecular and functional characterization of a c-type lysozyme from the Asian corn borer, Ostrinia furnacalis.

Project description:Some lepidopteran lysozymes have been reported to display activity against Gram-positive and Gram-negative bacteria, in contrast to most lysozymes that are active only against Gram-positive bacteria. OstrinLysC, a c-type lysozyme, was purified from the Asian corn borer, Ostrinia furnacalis Guenée (Lepidoptera: Pyralidae), and shows activity against Gram-positive and Gram-negative bacteria. The NH2-terminal amino acid sequence was determined by Edman degradation and used in a homology cloning strategy. The gene coding for OstrinLysC contains three exons and two introns. The expression profile of the OstrinlysC gene was examined by quantitative real-time PCR. Following injection of the larvae with bacteria, the OstrinlysC gene is strongly up-regulated in immune tissues. Transcripts were also detected in gut tissue. After feeding the larvae with bacteria, OstrinlysC transcripts increased in immune tissues. A very low level of transcript abundance was also detected in gut tissue. These results suggested that the OstrinlysC gene is involved in immune responses. The three dimensional structure of OstrinLysC was predicted. Based on comparison of the 3-D structure of OstrinLysC with that of silkworm lysozyme and chicken lysozyme, we hypothesize that the positive charge-rich surface and the short loop-2, which is close to the cluster of hydrophobic residues, may play important roles in the interaction with the outer membrane of Gram-negative bacterial cell walls.

| S-EPMC3011828 | biostudies-literature

Antifeedant and ovicidal activities of ginsenosides against Asian corn borer, Ostrinia furnacalis (Guenee).

Project description:INTRODUCTION:Ginsenosides, including protopanaxdiol (PPD) and protopanaxtriol (PPT) type ginsenosides, have been identified as natural insecticidess. This study aimed to investigate the antifeedant and ovicidal activities of total ginsenosides, protopanaxdiol saponins (PDS) and protopanaxtriol saponins (PTS) against Asian corn borer, O. furnacalis (Guenee). METHODS AND RESULTS:O. furnacalis egg masses (> 40 eggs) at 0-, 1- and 2-day-old were dipped into ginsenosides and egg hatchability was significantly inhibited by total ginsenosides, PDS, and PTS in dose and egg-age dependent manners. 100 mg/ml PDS had the strongest ovicidal activity against 0- (80.58 ± 0.95%), 1- (71.48 ± 5.70%), and 2-day-old eggs (64.31 ± 3.20%). In no-choice and choice feeding tests, we observed that the 3rd instar larvae consumed decreased area of leaves treated with ginsenosides, and the antifeedant activities of total ginsenosides, PDS, and PTS against the 3rd instar larvae were time and dose-dependent, with higher activities at 48 h. 100 mg/ml PDS had relative higher antifeedant activity (88.39 ± 3.43% in no-choice and 80.9±4.36% in choice) than total ginsenosides and PTS at all time intervals, except at 48 h in no-choice test. In further experiments, we found PPD ginsenosides (Rb1, Rb2, Rc, and Rd) had relative higher time and dose dependent antifeedant activities than PPT ginsenosides (Re and Rg1). CONCLUSIONS:Our results suggested the insecticidal action of total ginsenosides, PDS, and PTS on O. furnacalis. All ginsenosides, especially PDS, showed antifeedant and ovicidal activities against O. furnacalis.

| S-EPMC6377112 | biostudies-literature

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