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molecular cloning and expression of bacillus thuringiensis subsp. galleriae insecticidal crystal protein genes in escherichia coli.the location of the toxin gene of b. thuringiensis subsp. galleriae (h5ab) on the mr-130md plasmid is determined by molecular cloning. double digestion fragments (bamhi and sali) and psti restriction fragments as well, from the 130 md plasmid, of b. thuringiensis subsp. galleriae, are ligated with the cloning vector pat 153 respectively and transformed into e. coli strain hb 101. out of 208 transformants, three colonies (fg2, fg9, fg19) give positive hybridization reaction using the hd-1 delta-e ...19892554939
evaluation of transgenic bt corn for resistance to the asian corn borer (lepidoptera: pyralidae).the asian corn borer, ostrinia furnacalis (guenée), is the most important insect pest on corn in china. bt transgenic corn provides a new tool for asian corn borer control. monsanto's yieldgard bt transgenic corn expressing cry1ab protein, and a non-bt control, were evaluated in beijing. laboratory bioassays were carried out by exposing neonates to an agar-free diet containing bt corn whorl leaves, tassels, and anthers, or by exposing neonates directly to fresh silk and pollen. these are the tis ...200312852639
molecular genetics and evolution of pheromone biosynthesis in lepidoptera.a great diversity of pheromone structures are used by moth species (insecta: lepidoptera) for long-distance mating signals. the signal/response channel seems to be narrow for each species, and a major conundrum is how signal divergence has occurred in the face of strong selection pressures against small changes in the signal. observations of various closely related and morphologically similar species that use pheromone components biosynthesized by different enzymes and biosynthetic routes unders ...200312876197
identification of cry1i-type genes from bacillus thuringiensis strains and characterization of a novel cry1i-type gene.a pcr-restriction fragment length polymorphism method for identification of cry1i-type genes from bacillus thuringiensis was established by designing a pair of universal primers based on the conserved regions of the genes to amplify 1,548-bp cry1i-type gene fragments. amplification products were digested with the bsp119i and bani enzymes, and four kinds of known cry1i-type genes were successfully identified. the results showed that cry1i-type genes appeared in 95 of 115 b. thuringiensis isolates ...200312957903
molecular genetics and evolution of pheromone biosynthesis in lepidoptera.a great diversity of pheromone structures are used by moth species (insecta: lepidoptera) for long-distance mating signals. the signal/response channel seems to be narrow for each species, and a major conundrum is how signal divergence has occurred in the face of strong selection pressures against small changes in the signal. observations of various closely related and morphologically similar species that use pheromone components biosynthesized by different enzymes and biosynthetic routes unders ...200316576755
novel sex pheromone desaturases in the genomes of corn borers generated through gene duplication and retroposon fusion.the biosynthesis of female moth sex pheromone blends is controlled by a number of different enzymes, many of which are encoded by members of multigene families. one such multigene family, the acyl-coa desaturases, is composed of certain genes that function as key players in moth sex pheromone biosynthesis. although much is known regarding the function of some of these genes, very little is known regarding how novel genes have evolved within this family and how this might impact the establishment ...200717360547
[expression and insecticidal activity of a novel gene cry2ab4 from bacillus thuringiensis strain b-pr-88].the full length cry2ab gene was cloned by pcr-rflp method from bt strain b-pr-88, which was isolated in china with high toxicity to the lepidopteran insect pests. nucleic acid sequence analysis showed that this gene was 1902 base pairs encoding 633 amino acids. this cry gene was named cry2ab4 as a novel gene by bacillus thuringiensis delta endotoxin nomenclature committee. the full open reading frame sequence of the cry2ab4 gene was amplified with a pair of pcr primers l2ab5/l2ab3 designed accor ...200717822035
sex-specific death in the asian corn borer moth (ostrinia furnacalis) infected with wolbachia occurs across larval development.maternally inherited endosymbiotic bacteria of the genus wolbachia induce various kinds of reproductive alterations in their arthropod hosts. in a wolbachia-infected strain of the adzuki bean borer moth, ostrinia scapulalis (lepidoptera: crambidae), males selectively die during larval development, while females selectively die when wolbachia are eliminated by antibiotic treatment. we found that naturally occurring wolbachia in the congener o. furnacalis caused sex-specific lethality similar to t ...200717893742
cloning and characterization of a novel cry1a toxin from bacillus thuringiensis with high toxicity to the asian corn borer and other lepidopteran insects.a novel cry1a was cloned from bacillus thuringiensis strain bt8 and expressed in the b. thuringiensis acrystalliferous mutant hd73(-). the gene, designated cry1ah1, encoded a protein with a molecular weight of 134 kda. reverse transcriptase-pcr and western blotting showed that cry1ah was expressed in the host strain bt8. the toxin expressed in hd73(-) exhibited high toxicity against lepidopteran larvae of ostrinia furnacalis, helicoverpa armigera, chilo suppressalis, and plutella xylostella. the ...200818248430
molecular and functional characterization of a c-type lysozyme from the asian corn borer, ostrinia furnacalis.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 str ...200919613460
[bt toxic protein expression in insect-resistant transgenic corns and its transfer to and accumulation in ostrinia furnacalis].taking the insect-resistant transgenic corn varieties g03-2396 and g03-2739 and the conventional corn variety suyu 16 as test materials, a bioassay in laboratory was conducted to test their resistance against ostrinia furnacalis. the bt toxin expression in different tissues of the two transgenic corns, the ingestion of bt toxin by the 3rd and 5th instar of o. furnacalis, and the bt toxin amount in feces of o. furnacalis larvae fed with bt corns were analyzed by using enzyme-linked-immunosorbent- ...200920136015
identification of immunity-related genes in ostrinia furnacalis against entomopathogenic fungi by rna-seq analysis.the asian corn borer (ostrinia furnacalis (guenée)) is one of the most serious corn pests in asia. control of this pest with entomopathogenic fungus beauveria bassiana has been proposed. however, the molecular mechanisms involved in the interactions between o. furnacalis and b. bassiana are unclear, especially under the conditions that the genomic information of o. furnacalis is currently unavailable. so we sequenced and characterized the transcriptome of o. furnacalis larvae infected by b. bass ...201424466095
identification, characterization, and expression of a novel p450 gene encoding cyp6ae25 from the asian corn borer, ostrinia furnacalis.an allele of the cytochrome p450 gene, cyp6ae14, named cyp6ae25 (genbank accession no. eu807990) was isolated from the asian com borer, ostrinia fumacalis (guenée) (lepidoptera: pyralidae) by rt-pcr. the cdna sequence of cyp6ae25 is 2315 bp in length and contains a 1569 nucleotides open reading frame encoding a putative protein with 523 amino acid residues and a predicted molecular weight of 59.95 kda and a theoretical pi of 8.31. the putative protein contains the classic heme-binding sequence m ...021529257
cloning, expression, and characterization of prophenoloxidases from asian corn borer, ostrinia furnacalis (gunée).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 ...201628078308
transcriptome comparison analysis of ostrinia furnacalis in four developmental stages.the asian corn borer, ostrinia furnacalis, is one of the most destructive pests of maize and causes huge losses in maize yield each year. in order to characterize the different developmental stages, a high-throughput sequencing platform was employed to perform de novo transcriptome assembly and gene expression analysis for the egg, larva, pupa and adult stages. approximately 185 million reads were obtained, trimmed, and assembled into 42,638 unigenes with an average length of 801.94 bp and an n5 ...201627713521
molecular cloning, characterization and expression profiling of a ryanodine receptor gene in asian corn borer, ostrinia furnacalis (guenée).ryanodine receptor (ryr) ca(2+) release channel is the target of diamide insecticides, which show selective insecticidal activity against lepidopterous insects. to study the molecular mechanisms underlying the species-specific action of diamide insecticides, we have cloned and characterized the entire cdna sequence of ryr from ostrinia furnacalis (named as ofryr). the ofryr mrna has an open reading frame of 15324 bp nucleotides and encodes a 5108 amino acid polypeptide that displays 79-97% ident ...201324098400
identification and expression profile analysis of antimicrobial peptide/protein in asian corn borer, ostrinia furnacalis (guenée).antimicrobial peptides/proteins (amps) are a group of immune proteins that exhibit strong antibiotic properties against numerous infectious bacterial strains. they are evolutionarily conserved and present in every kingdom and phylum, ranging from prokaryotes to humans. we analyzed the transciptome from the larvae of asian corn borer, ostrinia furnacalis (guenée), and identified several putative amp transcripts, ofglys5, ofglys6, ofglys10, ofgatt, and ofgiid. ofglys5, ofglys6, and ofglys10 are al ...201324155672
de novo transcriptome and expression profile analyses of the asian corn borer (ostrinia furnacalis) reveals relevant flubendiamide response genes.the asian corn borer (acb), ostrinia furnacalis (guenée), has become the most damaging insect pest of corn in asia. however, the lack of genome or transcriptome information heavily hinders our further understanding of acb in every aspect at a molecular level and on a genome-wide scale. here, we used the ion torrent personal genome machine (pgm) sequencer to explore the acb transcriptome and to identify relevant genes in response to flubendiamide, showing high selective activity against acb.201728056803
inheritance patterns, dominance and cross-resistance of cry1ab- and cry1ac-selected ostrinia furnacalis (guenée).two colonies of asian corn borer, ostrinia furnacalis (guenée), artificially selected from a bt-susceptible colony (acb-bts) for resistance to cry1ab (acb-abr) and cry1ac (acb-acr) toxins, were used to analyze inheritance patterns of resistance to cry1 toxins. acb-abr and acb-acr evolved significant levels of resistance, with resistance ratios (rr) of 39-fold and 78.8-fold to cry1ab and cry1ac, respectively. the susceptibility of acb-abr larvae to cry1ac and cry1f toxins, which had not previousl ...201425216083
de novo transcriptome analysis of wing development-related signaling pathways in locusta migratoria manilensis and ostrinia furnacalis (guenée).orthopteran migratory locust, locusta migratoria, and lepidopteran asian corn borer, ostrinia furnacalis, are two types of insects undergoing incomplete and complete metamorphosis, respectively. identification of candidate genes regulating wing development in these two insects would provide insights into the further study about the molecular mechanisms controlling metamorphosis development. we have sequenced the transcriptome of o. furnacalis larvae previously. here we sequenced and characterize ...201425207539
genetic basis of cry1f-resistance in a laboratory selected asian corn borer strain and its cross-resistance to other bacillus thuringiensis toxins.the asian corn borer (acb), ostrinia furnacalis (guenée) (lepidoptera: crambidae), is the most destructive insect pest of corn in china. susceptibility to the cry1f toxin derived from bacillus thuringiensis has been demonstrated for acb, suggesting the potential for cry1f inclusion as part of an insect pest management program. insects can develop resistance to cry toxins, which threatens the development and use of bt formulations and bt crops in the field. to determine possible resistance mechan ...201627518813
homology of olfactory receptor neuron response characteristics inferred from hybrids between asian and european corn borer moths (lepidoptera: crambidae).first generation hybrid males from crosses between the asian corn borer (acb), ostrinia furnacalis, and the "univoltine z-strain" european corn borer (ecb), ostrinia nubilalis, were examined with respect to behavioral and physiological responses to acb and ecb pheromones. the hybrid males often flew to the pheromone of ecb z-strain, but very rarely to the acb pheromone. we mapped the tuning profiles of each orn of the f(1) hybrids with respect to the relevant pheromone components and a common be ...201019778540
altered olfactory receptor neuron responsiveness in rare ostrinia nubilalis males attracted to the o. furnacalis pheromone blend.three percent of e-strain ostrinia nubilalis males fly upwind in response to the ostrinia furnacalis pheromone blend [a 40:60 ratio of (e)-12-tetradecenyl acetate to (z)-12-tetradecenyl acetate (e12-14:oac to z12-14:oac)], in addition to their own pheromone blend [a 99:1 ratio of (e)-11-tetradecenyl acetate to (z)-11-tetradecenyl acetate) (e11-14:oac to z11-14:oac)]. we assessed the olfactory receptor neuron (orn) responses of these behaviorally "rare" males versus those of normal males. for the ...200717624366
evidence of olfactory antagonistic imposition as a facilitator of evolutionary shifts in pheromone blend usage in ostrinia spp. (lepidoptera: crambidae).olfactory receptor neuron (orn) response was measured to assess why some males ("rare males") of the asian corn borer (acb), ostrinia furnacalis, have a broad behavioral response to fly upwind to both the acb and the european corn borer (ecb), ostrinia nubilalis, pheromone blends. we performed single-sensillum electrophysiological recordings on acb males that had been behaviorally assessed for upwind flight response to the acb blend [60:40 (z)-12-tetradecenyl acetate (z12-14:oac) to (e)-12-tetra ...200717374381
more rare males in ostrinia: response of asian corn borer moths to the sex pheromone of the european corn borer.a previous flight tunnel study showed that 3-5% of european corn borer (ecb) moths, ostrinia nubilalis (z/e11-14:oac), could fly upwind and make contact with sources releasing the sex pheromone of the related asian corn borer (acb), ostrinia furnacalis (2:1 z/e12-14:oac). in this study, we show that rare males (3-4%) are also present in south korean acb that respond to the sex pheromone blends of the ecb uz (97:3 z/e11-14:oac) and be (1:99 z/e11-14:oac) pheromone races. we also show that the upw ...200717146720
distribution and variations of three 1,4-benzoxazin-3-ones in maize induced by the asian corn borer, ostrinia furnacalis (guenée).contents of three 1,4-benzoxazin-3-ones in tissue samples from different parts (young leaf, second leaf, old leaf, stem and root) of young maize plants of 4-leaves stage, fed by the third instar larvae of the asian corn borer, ostrinia furnacalis (guenée), were analyzed by high-performance liquid chromatography-mass spectroscopy (hplc-ms). samples were taken immediately (set a) or 48 h (set b) after larvae had fed on the second leaf for 48 h. the three 1,4-benzoxazin-3-ones investigated in our e ...200616729586
silent genes and rare males: a fresh look at pheromone blend response specificity in the european corn borer moth, ostrinia nubilalis.the response of male moths from two pheromone races of the european corn borer, ostrinia nubilalis, was measured in a flight tunnel assay to different ratios of structurally different compounds that comprise the sex pheromone of the asian corn borer, ostrinia furnacalis. for both o. nubilalis races, between 1 and 5% of the males completed upwind flights to two different blends of the o. furnacalis pheromone components (the 2:1 z/e12-14:oac female-produced blend, and a 97:3 z/e mix), confirming t ...200315841231
genetic variation and phylogenetic relationships among six populations of corn borers in china.the genetic variation among and within six populations of the corn borer was determined by using random amplified polymorphic dna (rapd) markers. extensive genetic variability was detected. of the 802 rapd markers obtained, 781 (97.4%) were polymorphic among populations. genetic similarities and distances between each pair of individuals were calculated. upgma cluster analysis showed that the yn population (ostrinia nubilalis hübner) and the other five populations (ostrinia furnacalis guenée) ma ...19989791723
the cultivation of bt corn producing cry1ac toxins does not adversely affect non-target arthropods.transgenic corn producing cry1ac toxins from bacillus thuringiensis (bt) provides effective control of asian corn borer, ostrinia furnacalis (guenée), and thus reduces insecticide applications. however, whether bt corn exerts undesirable effects on non-target arthropods (ntas) is still controversial. we conducted a 2-yr study in shangzhuang agricultural experiment station to assess the potential impact of bt corn on field population density, biodiversity, community composition and structure of n ...201425437213
downregulation and mutation of a cadherin gene associated with cry1ac resistance in the asian corn borer, ostrinia furnacalis (guenée).development of resistance in target pests is a major threat to long-term use of transgenic crops expressing bacillus thuringiensis (bt) cry toxins. to manage and/or delay the evolution of resistance in target insects through the implementation of effective strategies, it is essential to understand the basis of resistance. one of the most important mechanisms of insect resistance to bt crops is the alteration of the interactions between cry toxins and their receptors in the midgut. a cry1ac-selec ...201425216082
screening of cry-type promoters with strong activity and application in cry protein encapsulation in a sigk mutant.to optimize the expression of cry genes in a bacillus thuringiensis sigk mutant failing in crystal releasing, the transcriptional activity of the cry promoters cry1a, cry3a, cry4a, and cry8e was compared using lacz gene fusions. a beta-galactosidase assay indicated that the cry8e promoter showed the highest transcriptional activity. a novel escherichia coli-b. thuringiensis shuttle vector pht315-8e21b was constructed for cry gene expression using the cry8e promoter and the multiple cloning sites ...201424928660
male- and female-biased gene expression of olfactory-related genes in the antennae of asian corn borer, ostrinia furnacalis (guenée) (lepidoptera: crambidae).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 in ...201526062030
active-pocket size differentiating insectile from bacterial chitinolytic +¦-n-acetyl-d-hexosaminidases.chitinolytic +¦-n-acetyl-d-hexosaminidase is a branch of the gh20 (glycoside hydrolase family 20) +¦-n-acetyl-d-hexosaminidases that is only distributed in insects and micro-organisms, and is therefore a potential target for the action of insecticides. pugnac [o-(2-acetamido-2-deoxy-d-glucopyransylidene)-amino-n-phenylcarbamate] was initially identified as an inhibitor against gh20 +¦-n-acetyl-d-hexosaminidases. so far no crystal structure of pugnac in complex with any gh20 +¦-n-acetyl-d-hexosam ...201121692744
sex pheromone desaturase functioning in a primitive ostrinia moth is cryptically conserved in congeners' genomes.(e)-11- and (z)-11-tetradecenyl acetate are the most common female sex pheromone components in ostrinia moths. the ?11-desaturase expressed in the pheromone gland (pg) of female moths is a key enzyme that introduces a double bond into pheromone molecules. a single ?11-desaturase of ostrinia nubilalis, onubz/e11, has been shown to produce an ~7:3 mixture of (e)-11- and (z)-11-tetradecenoate from the substrate tetradecanoate. in contrast, the sex pheromone of ostrinia latipennis, a primitive speci ...201121444802
structural determinants of an insect beta-n-acetyl-d-hexosaminidase specialized as a chitinolytic enzyme.β-n-acetyl-d-hexosaminidase has been postulated to have a specialized function. however, the structural basis of this specialization is not yet established. ofhex1, the enzyme from the asian corn borer ostrinia furnacalis (one of the most destructive pests) has previously been reported to function merely in chitin degradation. here the vital role of ofhex1 during the pupation of o. furnacalis was revealed by rna interference, and the crystal structures of ofhex1 and ofhex1 complexed with tmg-chi ...201021106526
variation in courtship ultrasounds of three ostrinia moths with different sex pheromones.moths use ultrasounds as well as pheromones for sexual communication. in closely related moth species, variations in ultrasounds and pheromones are likely to profoundly affect mate recognition, reproductive isolation, and speciation. the european corn borer, ostrinia nubilalis, and its asian congeners, ostrinia furnacalis and ostrinia scapulalis, exhibit within-species and between-species variation in their pheromone communication. recently, we reported ultrasound communication in o. furnacalis; ...201020957230
expression, purification and characterization of the chitinolytic beta-n-acetyl-d-hexosaminidase from the insect ostrinia furnacalis.insect beta-n-acetyl-d-hexosaminidases are of particular interest due to their multiple physiological roles in many life processes. chitinolytic beta-n-acetyl-d-hexosaminidases, which function only in chitin degradation in insects, have long been regarded as species-specific target potentials in developing environmental friendly pesticides. here the chitinolytic beta-n-acetyl-d-hexosaminidase from the insect ostrinia furnacalis was cloned and expressed in the yeast strain, pichia pastoris, to me ...200919520169
asian corn borer (acb) and non-acb pests in gm corn (zea mays l.) in the philippines.the asian corn borer (acb), ostrinia furnacalis (guenée), has become the most damaging pest in corn in south-east asia. corn farmers in the philippines have incurred great yield losses in the past decades because of acb infestation. bacillus thuringiensis (bt) and bt herbicide-tolerant (btht) corns have been developed to reduce borer attacks worldwide. this study assessed the extent of acb and non-acb pest infestations in both gm and non-gm corn in isabela province, the philippines. specific aim ...201323401215
serine proteases sp1 and sp13 mediate the melanization response of asian corn borer, ostrinia furnacalis, against entomopathogenic fungus beauveria bassiana.exposure to entomopathogenic fungi is one approach for insect pest control. little is known about the immune interactions between fungus and its insect host. melanization is a prominent immune response in insects in defending against pathogens such as bacteria and fungi. clip domain serine proteases in insect plasma have been implicated in the activation of prophenoloxidase, a key enzyme in the melanization. the relationship between host melanization and the infection by a fungus needs to be est ...201525900291
a novel beta-n-acetyl-d-hexosaminidase from the insect ostrinia furnacalis (guenée).exploiting specific targets is of specific interest in developing eco-friendly pesticides. we isolated, purified and characterized a novel beta-n-acetyl-d-hexosaminidase (ofhex1) from the fifth instar larva integument of the asian corn borer, ostrinia furnacalis (guenée). ofhex1 was purified 1468-fold to homogeneity with an activity yield of 20% by four column chromatography steps. under denaturing conditions, the molecular mass of ofhex1 was determined to be 67.0 kda by ms and sds/page, but 128 ...200818959754
proteomics of methyl jasmonate induced defense response in maize leaves against asian corn borer.jasmonic acid (ja) and methyl jasmonate (meja) regulate plant development, resistance to stress, and insect attack by inducing specific gene expression. however, little is known about the mechanism of plant defense against herbivore attack at a protein level. using a high-resolution 2-d gel, we identified 62 meja-responsive proteins and measured protein expression level changes.201525885025
effects of bacillus thuringiensis toxin cry1ac and beauveria bassiana on asiatic corn borer (lepidoptera: crambidae).in this study, interactions between cry1ac, a toxic crystal protein produced by bacillus thuringiensis (berliner), and beauveria bassiana on the mortality and survival of ostrinia furnacalis was evaluated in the laboratory. the results showed that cry1ac is toxic to o. furnacalis. not only were larval growth and development delayed, but pupation, pupal weight and adult emergency also decreased when larvae were fed on artificial diet containing purified cry1ac toxin. when third instars o. furnaca ...200818656480
olfactory neuron responsiveness and pheromone blend preference in hybrids between ostrinia furnacalis and ostrinia nubilalis (lepidoptera: crambidae).the olfactory receptor neuron (orn) and behavioral responses of hybrids between the asian corn borer (acb), ostrinia furnacalis, and the e-strain european corn borer (ecb(e)), ostrinia nubilalis were examined and compared to the parental populations. in hybrids and both parents, the large-spike-size orn was capable of responding to all four pheromone components of acb and ecb, despite differences in which compounds elicited the greatest spike frequency in each population. there was a small-spiki ...200818634788
production of n-acetyl-d-glucosamine from mycelial waste by a combination of bacterial chitinases and an insect n-acetyl-d-glucosaminidase.n-acetyl-d-glucosamine (glcnac) has great potential to be used as a food additive and medicine. the enzymatic degradation of chitin-containing biomass for producing glcnac is an eco-friendly approach but suffers from a high cost. the economical efficiency can be improved by both optimizing the member and ratio of the chitinolytic enzymes and using new inexpensive substrates. to address this, a novel combination of bacterial and insect chitinolytic enzymes was developed in this study to efficient ...201627546481
structural characteristics of an insect group i chitinase, an enzyme indispensable to moulting.insects possess a greater number of chitinases than any other organisms. this work is the first report of unliganded and oligosaccharide-complexed crystal structures of the insect chitinase ofchti from ostrinia furnacalis, which is essential to moulting. the obtained crystal structures were solved at resolutions between 1.7 and 2.2 å. a structural comparison with other chitinases revealed that ofchti contains a long substrate-binding cleft similar to the bacterial chitinase smchib from serratia ...201424699639
molecular characters and expression analysis of the gene encoding eclosion hormone from the asian corn borer, ostrinia furnacalis.using rapid amplification of cdna ends (race), the cdna encoding eclosion hormone (eh) was cloned from the brain of ostrinia furnacalis. the full osf-eh cdna is 986 bp and contains a 267 bp open reading frame encoding an 88 amino acid preprohormone, which including a hydrophobic 26 amino acid signal peptide and a 62 amino acid mature peptide. the mature osf-eh shows high identity with manduca sexta (95.2%), helicoverpa armigera (91.9%) and bombyx mori (85.5%), but low identify with tribolium cas ...200817852339
feminizing wolbachia in an insect, ostrinia furnacalis (lepidoptera: crambidae).wolbachia, which forms a group of maternally inherited bacteria in arthropods, often cause reproduction alterations in their hosts, such as cytoplasmic incompatibility, parthenogenesis, male-killing, hybrid breakdown and feminization. to date, wolbachia-induced feminization has been reported only in isopods. here we report that a wolbachia strain feminizes an insect host, ostrinia furnacalis. among 79 wild females of o. furnacalis examined, wolbachia infection was detected in 13 females. twelve ...200212180086
development of bt rice and bt maize in china and their efficacy in target pest control.rice and maize are important cereal crops that serve as staple foods, feed, and industrial material in china. multiple factors constrain the production of both crops, among which insect pests are an important one. lepidopteran pests cause enormous yield losses for the crops annually. in order to control these pests, china plays an active role in development and application of genetic engineering (ge) to crops, and dozens of ge rice and ge maize lines expressing insecticidal proteins from the soi ...201627763554
comparative binding of cry1ab and cry1f bacillus thuringiensis toxins to brush border membrane proteins from ostrinia nubilalis, ostrinia furnacalis and diatraea saccharalis (lepidoptera: crambidae) midgut tissue.the european (ostrinia nubilalis hübner) and asian corn borers (ostrinia furnacalis guenée) are closely related and display similar sensitivity to cry1 toxins. in this study, we compared the binding patterns of cry1ab and cry1f toxins between both ostrinia spp., as well as the expression of putative cadherin- and aminopeptidase-n (apn)-like protein receptors. additionally, cdna sequences of these putative toxin receptors from both ostrinia species were compared. ligand blots for both species ind ...201323999243
molecular characterization of glutaredoxin 2 from ostrinia furnacalis.glutaredoxins (grxs) play very important roles in maintaining intracellular redox homeostasis. in the present study, the full-length cdna sequence encoding grx2, named ofurgrx2 (genbank accession no. gu393246), was obtained from ostrinia furnacalis, using reverse transcription polymerase chain reaction and rapid amplification of cdna ends. sequence analysis revealed that the open reading frame of ofurgrx2 consists of 351 nucleotides encoding 116 amino acid residues with a predicted molecular wei ...201223621469
A modeling study for structure features of ß-N-acetylhexosaminidase from Ostrinia furna and its novel inhibitor allosamidin: species selectivity and multi-target characteristics.Insect ß-N-acetylhexosaminidase, a chitin degrading enzyme, is physiologically important during the unique life cycle of the insect. OfHex1, a ß-N-acetylhexosaminidase from the insect, Ostrinia furna, which was obtained by our lab (Gen Bank No.: ABI81756.1), was studied by molecular modeling as well as molecular docking with its inhibitor, allosamidin. 3D model of OfHex1 was built through the ligand-supported homology modeling approach. The binding modes of its substrate and inhibitor were propo ...201122177554
an improved pcr-restriction fragment length polymorphism (rflp) method for the identification of cry1-type genes.the cry1-type genes of bacillus thuringiensis represent the largest cry gene family, which contains 50 distinct holotypes. it is becoming more and more difficult to identify cry1-type genes using current methods because of the increasing number of cry1-type genes. in the present study, an improved pcr-restriction fragment length polymorphism (pcr-rflp) method which can distinguish 41 holotypes of cry1-type genes was developed. this improved method was used to identify cry1-type genes in 20 b. th ...201323995930
permanent genetic resources: isolation and characterization of microsatellite loci from the european corn borer, ostrinia nubilalis (hübner) (insecta: lepidoptera: crambidae).few useful microsatellites are available for population studies of the european corn borer, ostrinia nubilalis (hübner). an enrichment strategy was used to develop microsatellite markers for o. nubilalis, and over 500 positive clones were isolated. seventy-five contained unique microsatellites, 10 of which were polymorphic with discernable polymerase chain reaction products. the 10 loci were surveyed for variability in 72 wild individuals from central iowa. five loci showed no deviation from har ...200821585805
molecular characterization of a short peptidoglycan recognition protein (pgrp-s) from asian corn borer (ostrinia furnacalis) and its role in triggering propo activity.peptidoglycan recognition proteins (pgrps) are non-specific immune molecules of insects, and vertebrates etc., but are not present in plants and nematodes. in the current experiment, a pgrp dna sequence (2,910 bp containing four exons) was identified from genomic dna library of asian corn borer, ostrinia furnacalis, and a full-length cdna programming pgrp was cloned (designed as ofpgrp-s) with an open reading frame of 579 bp, having 192 amino acid. this inferred amino acid sequence showed maximu ...201423907718
purification and characterization of hemolymph prophenoloxidase from ostrinia furnacalis (lepidoptera: pyralidae) larvae.prophenoloxidase (ppo) was isolated from the hemolymph of ostrinia furnacalis larvae and purified to homogeneity. a 369.85-fold purification and 35.34% recovery of activity were achieved by employing ammonium sulfate precipitation, blue sepharose cl-6b chromatography and phenyl sepharose cl-4b chromatography. the purified enzyme exhibits a band with a molecular mass of 158 kda on native page and two spots with a molecular mass of 80 kda and a pi of 5.70, and a molecular mass of 78 kda and a pi o ...200818577463
the gene, expression pattern and subcellular localization of chitin synthase b from the insect ostrinia furnacalis.insect midgut peritrophic membrane (pm) is a functional structure that protects insects against chemical damage and microorganism infection. the essential component in pm is chitin and its synthesis is catalyzed by class b chitin synthase (chsb), which plays a unique role in chitin-containing organisms and thus represents a potential target for eco-friendly pesticides. cdna and gdna of chsb from a widely spread pest ostrinia furnacalis (ofchsb) were obtained and their sequences and transcription ...201021145310
use of a pooled clone method to isolate a novel bacillus thuringiensis cry2a toxin with activity against ostrinia furnacalis.a pooled clone method was developed to screen for cry2a genes. this metagenomic method avoids the need to analyse isolated bacillus thuringiensis strains by performing gene specific pcr on plasmid-enriched dna prepared from a pooled soil sample. using this approach the novel holotype gene cry2ah1 was cloned and characterized. the toxin gene was over-expressed in escherichia coli rosetta (de3) and the expressed toxin accumulated in both the soluble and insoluble fractions. the soluble cry2ah1 was ...201323711413
characterization of cry9da4, cry9eb2, and cry9ee1 genes from bacillus thuringiensis strain t03b001.three cry9 genes, cry9da4, cry9eb2, and cry9ee1, were cloned from bacillus thuringiensis strain t03b001 using a high-resolution melting analysis method. all three cry9 genes were overexpressed in escherichia coli rosetta (de3), and the expressed products cry9eb2 and cry9ee1 were shown to be toxic to plutella xylostella and ostrinia furnacalis, but not to helicoverpa armigera or colaphellus bowringi. the bioassay of cry9eb2 and cry9ee1 against cry1ac-resistant p. xylostella strains indicated that ...201323455566
physiological adaptation of the asian corn borer ostrinia furnacalis to chemical defenses of its host plant, maize.a number of gramineous plants such as maize contain cyclic hydroxamic acids (chx) that are toxic to many herbivores. among the ostrinia species found in japan, the asian corn borer ostrinia furnacalis is the only one that utilizes maize, a gramineous plant. we used o. furnacalis and two congeners, ostrinia scapulalis and ostrinia latipennis, to obtain insights into the physiological adaptation of o. furnacalis to chx. when an artificial diet containing a low concentration (0.3mg/g diet) of chx w ...201020435041
identification of an alkaline phosphatase as a putative cry1ac binding protein in ostrinia furnacalis (guenée).asian corn borer (acb), ostrinia furnacalis, is an important insect pest of maize susceptible to different cry1a toxins. based on amino acid sequence alignment of alp sequences from lepidopteran larvae an alp gene was cloned from acb, named ofalp. pull dawn assays using biotinylated cry1ac and brush border membrane vesicles isolated from second instar acb larvae showed that four proteins of 50, 65, 68 and 70kda precipitated with the cry1ac. the 65kda band cross-reacted with the anti-ofalp monocl ...201627265829
construction of transgenic trichoderma koningi with chit42 of metarhizium anisopliae and analysis of its activity against the asian corn borer.the chit42 gene was cloned from metarhizium anisopliae cy1 and was inserted into trichoderma koningii t30 genome by protoplast transformation. sixteen transgenic isolates were identified by polymerase chain reaction analysis. the chit42 gene was 1275 bp in length and its coded protein was approximately 42 kda in size. semi-quantitative rt-pcr analysis and the measurement of the chitinase activity under induced conditions were conducted. mortality of the asian corn borer (ostrinia furnacalis) was ...201222560024
cloning, expression, purification, and insecticidal activity of a novel cry1na3 toxin from bacillus thuringiensis brc-zyr2.bacillus thuringiensis produces a variety of insecticidal crystal proteins (icps). genome sequencing is a promising strategy for detecting and identifying b. thuringiensis icps, which are of great interest to the biocontrol field. in this study, a novel icp gene was cloned from b. thuringiensis brc-zyr2 based on genomic data from 454 gs-flx titanium sequencing and an analysis of the results using the b. thuringiensis toxin_scanner (http://bcam.hzaubmb.net/bttoxin_scanner/index.php). cry1na3 desi ...201627170729
does bt maize expressing cry1ac protein have adverse effects on the parasitoid macrocentrus cingulum (hymenoptera: braconidae)?the potential effects of insect-resistant, genetically engineered (ge) crops on non-target organisms, especially on predators and parasitoids, must be evaluated before their commercial cultivation. the effects of ge maize that produces cry1ac toxin on the parasitoid macrocentrus cingulum were assessed by direct bioassay and indirect bioassay. in the indirect bioassay, parasitism rate, cocoon weight and the number of m. cingulum progeny produced per host were significantly reduced when m. cingulu ...201627126195
identification of differentially expressed micrornas between bacillus thuringiensis cry1ab-resistant and -susceptible strains of ostrinia furnacalis.the asian corn borer (acb), ostrinia furnacalis (guenée), can develop strong resistance to cry1ab, the most widely commercialized cry toxin for bt maize worldwide. it is essential to understand the mechanism of resistance for management of this species, but information on the post-transcriptional regulation of bt resistance in this target insect is limited. in the present study, rna was extracted from the acb in various larval stages (1-5 instar) from cry1ab-sensitive (acb-bts) and -resistant (a ...201526486179
two kinds of sex ratio distorters in a moth, ostrinia scapulalis.we surveyed female-biased sex ratio traits (sr traits) in a moth, ostrinia scapulalis (lepidoptera: crambidae) from six geographic locations in japan. a small proportion (8.3%) of females showed the sr trait. about half of them were associated with a wolbachia infection (srw+), whereas the remaining ones were not (srw-). no significant differences in frequencies were detected among locations and years for either srw+ or srw- females. in srw+ matrilines, the sr trait was inherited. curing the wol ...200314663516
conserved rna cis-elements regulate alternative splicing of lepidopteran doublesex.doublesex (dsx) is a downstream key regulator in insect sex determination pathway. in drosophila, alternative splicing of dm-dsx gene is sex-specifically regulated by transformer (tra), in which the functional tra promotes female-specific dm-dsx. however, the sex determination pathway in lepidoptera is not well understood; here we focused on alternative splicing of doublesex (dsx) in two agricultural pests, asian corn borer (ostrinia furnacalis) and cotton bollworm (helicoverpa armigera), as wel ...201424239545
genetic transformation mediated by piggybac in the asian corn borer, ostrinia furnacalis (lepidoptera: crambidae).the asian corn borer, ostrinia furnacalis, is a serious pest of corn, sorghum, and cotton in china and other asian countries. the present study is the first attempt to establish the transgenic line in o. furnacalis using a piggybac transposon, which will shed light on the future genetic control of o. furnacalis. a piggybac vector pbac[a3egfp] was constructed to express enhanced green fluorescence protein (egfp)under the control of bombyx mori actin3 promoter. transient egfp expression was detect ...201222696097
physiological significance of alternatively spliced exon combinations of the single-copy gene class a chitin synthase in the insect ostrinia furnacalis (lepidoptera).insect chitin synthase is an essential enzyme involved in chitin biosynthesis in insects. chitin synthase a (chsa) is expressed in different insect tissues during different developmental stages. chsa contains alternative-splicing exons that allow tissue- and development-specific chitin synthesis. here, we report that ofchsa from the lepidopteran ostrinia furnacalis contains two alternative-splicing exons, exons 2a and 2b and exons 19a and 19b. although four combinations of these exons are theore ...201222607200
a novel alternative splicing site of class a chitin synthase from the insect ostrinia furnacalis - gene organization, expression pattern and physiological significance.insect chitin synthase a (chsa) catalyzes chitin biosynthesis in tissues that develop from ectoderm. since only one gene copy encodes chsa, we hypothesized that chsa is very likely to exist as isoforms through alternative splicing, and the functions of these isoforms may be tissue-specific. besides the known alternative splicing exons in the mid-orf region, we report here the alternative exons (ofchsa-2a and ofchsa-2b) of ofchsa, the chitin synthase a from the lepidopteran pest ostrinia furnacal ...201121933709
a new insect cell line from the pupal ovary of the asian corn borer moth ostrinia furnacalis.ovaries were removed from ostrinia furnacalis (guenée) pupae and were placed in a flask containing tnm-fh medium with 10% inactivated fetal bovine serum. cell migration occurred after about 1 wk of the initiation in june 2011. the migrated cells were distributed over most of the flask and were treated with n-methyl-n'-nitro-n-nitrosoguanidine (mnng), a chemical carcinogen, after about 1 mo of initiation for 26 d. cells were first subcultured successfully 12 d after the mnng was removed, followed ...201424163160
integrin β1 subunit from ostrinia furnacalis hemocytes: molecular characterization, expression, and effects on the spreading of plasmatocytes.when lepidopteran larvae are infected by a large quantity of pathogens or parasitized by nonadaptive parasitoids, hemocytes in the hemocoel will encapsulate these foreign invaders. cellular encapsulation requires hemocytes, particularly plasmatocytes, to change their states from nonadhesive, spherical cells into adhesive, spreading cells. however, it is unclear how the changes of plasmatocytes are regulated. here we report that the integrin β1 subunit from hemocytes of ostrinia furnacalis (ofint ...201020708011
cloning, expression and characterization of ostrinia furnacalis serpin1, a regulator of the prophenoloxidase activation system.serine protease inhibitors of the serpin superfamily are regulators of proteases involved in a variety of physiological processes including immune responses. in this study, we have isolated a full-length serpin cdna from ostrinia furnacalis. the 1188 bp open reading frame encodes a 395-residue protein with a theoretical molecular mass of 43.3 kda and an isoelectric point of 4.92. ofserpin1 contains a putative signal peptide followed by a conserved domain including a reactive center loop (rcl) wi ...201626589634
cloning, expression and biocharacterization of ofcht5, the chitinase from the insect ostrinia furnacalis.chitinase catalyzes β-1,4-glycosidic linkages in chitin and has attracted research interest due to it being a potential pesticide target and an enzymatic tool for preparation of n-acetyl-β-d-glucosamine. an individual insect contains multiple genes encoding chitinases, which vary in domain architectures, expression patterns, physiological roles and biochemical properties. herein, ofcht5, the glycoside hydrolase family 18 chitinase from the widespread lepidopteran pest ostrinia furnacalis, was cl ...201323955855
bt cry1ie toxin does not impact the survival and pollen consumption of chinese honey bees, apis cerana cerana (hymenoptera, apidae).the cry1ie gene may be a good candidate for the development of bt maize because over-expression of cry1ie is highly toxic to lepidopteran pests such as heliothis armigera hübner and ostrinia furnacalis guenée. the bt cry1ie gene also has no cross resistance with other insecticidal proteins such as cry1ab, cry1ac, cry1ah, or cry1f. chinese honey bees (apis cerana cerana) are potentially exposed to insect-resistant genetically modified (irgm) crops expressing cry1ie toxin via the collection of irg ...201627670271
hemomucin, an o-glycosylated protein on embryos of the wasp macrocentrus cingulum that protects it against encapsulation by hemocytes of the host ostrinia furnacalis.it is unclear how endoparasites passively evade their host's immune reactions in most parasite-host systems. hemomucin from the parasitoid wasp macrocentrus cingulum (mchem) is a 97-kda transmembrane protein containing 51 potential o-glycosylation sites that can be specifically recognized by arachis hypogaea lectin. mchem mrna is highly expressed in m. cingulum eggs, morulae and secondary embryos, and mchem protein is mainly located on the extraembryonic membrane of embryos. when secondary embry ...201424776378
[expression of cry1ac gene directed by pexsypromoter of the exsy gene encoding component protein of exosporium basal layer in bacillus thuringiensis].to discover new elements for cry gene expression, pexsy, which is the promoter of the exosporium basal layer structural gene exsy, was used to express cry1ac gene in bacillus thuringiensis.201425803890
identification of a new cry1i-type gene as a candidate for gene pyramiding in corn to control ostrinia species larvae.pyramiding of diverse cry toxin genes from bacillus thuringiensis with different modes of action is a desirable strategy to delay the evolution of resistance in the european corn borer (ostrinia nubilalis). considering the dependency of susceptibility to cry toxins on toxin binding to receptors in the midgut of target pests, a diverse mode of action is commonly defined as recognition of unique binding sites in the target insect. in this study, we present a novel cry1ie toxin gene (cry1ie2) as a ...201525795679
suitability of three ostrinia species as hosts for macrocentrus cingulum: a comparison of their encapsulation abilities.two cornborer species, ostrinia furnacalis (lepidoptera: crambidae) and o. nubilalis, are major corn pests in asia and europe, respectively. in both continents, the larval endoparasitoid macrocentrus cingulum (hymenoptera: braconidae) develops on another, closely related stemborer, o. scapulalis, which feeds on mugwort and other dicotyledons. m. cingulum also emerges from o. furnacalis in asia and o. nubilalis in north america, but not from o. nubilalis in europe. we assessed the ability of thre ...201423956040
characterization of four midgut aminopeptidase n isozymes from ostrinia furnacalis strains with different susceptibilities to bacillus thuringiensis.the full-length cdna of four ofapn aminopeptidases were cloned and sequenced from susceptible and resistant ostrinia furnacalis strains. four sequences were identified as apn because they shared the common structural features with apn from lepidoptera, including the signal peptide, gpi anchor signal, the zinc binding/gluzincin motif hex2hx18e and the gluzincin aminopeptidase motif gamen. compared with apn sequences from the susceptible strain, there were 9, 5, 10 and 12 amino acid variations in ...201424269376
impact on bacterial community in midguts of the asian corn borer larvae by transgenic trichoderma strain overexpressing a heterologous chit42 gene with chitin-binding domain.this paper is the first report of the impact on the bacterial community in the midgut of the asian corn borer (ostrinia furnacalis) by the chitinase from the transgenic trichoderma strain. in this study, we detected a change of the bacterial community in the midgut of the fourth instar larvae by using a culture-independent method. results suggested that proteobacteria and firmicutes were the most highly represented phyla, being present in all the midgut bacterial communities. the observed specie ...201323457472
captures of ostrinia furnacalis (lepidoptera: crambidae) with sex pheromone traps in ne china corn and soybeans.ostrinia furnacalis (guenée) (lepidoptera: crambidae), commonly referred to as the asian corn borer, is the most important corn pest in asia. although capturing males with pheromone traps has recently been the main monitoring tool and suppression technique, the best trap designs remain unclear. commercially available delta and funnel traps, along with laboratory-made basin and water traps, and modified delta traps, were evaluated in corn and soybean fields during 2013-2014 in ne china. the water ...201626362111
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