MicroRNAs (miRNAs) are endogenous 22-nt RNAs, which play important regulatory roles by post-transcriptional gene silencing. A computational strategy has been developed for the identification of conserved miRNAs base...MicroRNAs (miRNAs) are endogenous 22-nt RNAs, which play important regulatory roles by post-transcriptional gene silencing. A computational strategy has been developed for the identification of conserved miRNAs based on features of known metazoan miRNAs in red flour beetle (Tribolium castaneum), which is regarded as one of the major laboratory models of arthropods. Among 118 putative miRNAs, 47% and 53% of the predicted miRNAs from the red flour beetle are harbored by known protein-coding genes (intronic) and genes located outside (intergenic miRNA), respectively. There are 31 intronic miRNAs in the same transcriptional orientation as the host genes, which may share RNA polymerase II and spliceosomal machinery with their host genes for their biogenesis. A hypothetical feed-back model has been proposed based on the analysis of the relationship between intronic miRNAs and their host genes in the development of red flour beetle.展开更多
An allele of CYP6BQI3, named CYP6BQ 13v2 (GenBank accession no. FJ209361), was isolated from the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) by RT-PCR. The cDNA sequence of CYP6BQ...An allele of CYP6BQI3, named CYP6BQ 13v2 (GenBank accession no. FJ209361), was isolated from the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) by RT-PCR. The cDNA sequence of CYP6BQ13v2, 1 563 bp in length, contains an open reading frame of 1 554 nucleotides encoding a putative protein of 518 amino acid residues with a predicted molecular weight of 59.92 kDa and a theoretical pl of 7.60. The putative protein contains the classic hemebinding sequence motif F××G×××C×G (residues 456-465) conserved among all P450 enzymes as well as other characteristic motifs of all cytochrome P450s. It shares 98% identity with the previously published sequence of CYP6BQ13 (GenBank accession no. XP967146) from the T. castaneum genome project. Phylogenetic analysis of amino acid sequences from members of various P450 families indicated that there was closer phylogenetic relationship of CYP6BQ 13v2 with CYP302A1 and CYP307A1 mediating synthesis of the insect molting hormone, distant relationship with CYP6B1 metabolizing plant allelochemicals, CYP6D 1 linking to pyrethroid resistance and other members of CYP6 family. Expression test of the gene in the adults and immature stages of T. castaneum by quantitative real-time PCR revealed that CYP6BQ13v2 is expressed in all life stages investigated. The mRNA expression level in 1st instar larvae was 14.9- and 3.86-fold higher than those in pupae and adults, respectively. The CYP6BQ13v2 expression levels appeared in the order of 1st instar larvae, followed by 4th instar larvae, 7th instar larvae, adult, and pupae from high to low. The more bioinformation of CYP6BQ 13v2 was also analyzed.展开更多
The red flour beetle, Tribolium castaneum, is a major agriculture pest of stored grain, cereal products and peanuts for human consumption. It is reported that heat shock protein 18.3 of T. castaneum(Tchsp18.3) plays...The red flour beetle, Tribolium castaneum, is a major agriculture pest of stored grain, cereal products and peanuts for human consumption. It is reported that heat shock protein 18.3 of T. castaneum(Tchsp18.3) plays a significant role in stress resistance, development and reproduction. However, the regulatory systems of Tchsp18.3 remain unknown. Therefore, we compared the global transcriptome profiles of RNA interference(RNAi)-treated larvae(ds-Tchsp18.3) and control larvae of T. castaneum using RNA sequencing. Overall, we obtained 14 154 435 sequence reads aligned with 13 299 genes. Additionally, 569 differentially expressed genes(DEGs) were identified from the ds-Tchsp18.3 and control groups, of which 246 DEGs were annotated in the 47 Gene Ontology(GO) functional groups and 282 DEGs were assigned to 147 Kyoto Encyclopedia of Genes and Genomes(KEGG) biological signaling pathways. The DEGs encoding viperin, dorsal, Hdd11, PGRP2, defensin1 and defensin2 were simultaneously related to immunity and stress responses, which suggests that cross-talk might exist between the immunity and stress responses of T. castaneum. The knockdown of Tchsp18.3 gene expression suppressed the antioxidant activity process, which most likely modulated the effects of Tchsp18.3 on development and reproduction. Furthermore, the DEGs, including Blimp-1, Gld, Drm, Kinesin-14, Pthr2, Delta(11)-like and EGF-like domain protein 2, were also associated with the development and reproduction of ds-Tchsp18.3 insects. Additionally, knockdown of Tchsp18.3 amplified the serine protease(SP) signaling pathway to further regulate stress responses and innate immunity as well as development and reproduction of the red flour beetles. These results provide valuable insight into the molecular regulatory mechanism of Tchsp18.3 involved in insect physiology and further facilitate the research of suitable and sustainable management for pest control.展开更多
[Objeetive] The paper was to investigate the resistance of Tribolium castaneum to phosphine in Hainan Province. [Method] Using the measurement method of phosphine resistance of stored grain pests recommended by FAO, t...[Objeetive] The paper was to investigate the resistance of Tribolium castaneum to phosphine in Hainan Province. [Method] Using the measurement method of phosphine resistance of stored grain pests recommended by FAO, the phosphine resistance of eight strains of T. castaneum adults in Hainan Province was determined. [Result] The resistance coefficients of different strains of ?7. castaneum against phosphine ranged from 612 to 1 045, and all strains reached extremely high resistance level (Rf〉 160). When the concentration of phosphine was 200 mL/m^3, Rf612, Rf826.4 and Rfl 045 strains had complete lethal time of 8, 9 and 11 d, respectively. [Condusion] T. castarveum in Hainan Province had very strong resistance to phosphine, and the operation scheme of low concentration and long duration should be adopted in elimination of T. castaneum.展开更多
In order to elucidate the ability of stored product insects to resist high temperature at varying temperature conditions,the influence of different heating rates on the mortality of all life stages of Tribolium castan...In order to elucidate the ability of stored product insects to resist high temperature at varying temperature conditions,the influence of different heating rates on the mortality of all life stages of Tribolium castaneum(Herbst)(Coleoptera:Tenebrionidae)was investigated.After acclimation from 28 to 42,44 and 46℃ at different heating rates(0,1.25,2.50,5.00,and 10.00℃/h),eggs,larvae,pupae,and adults of T.castaneum were exposed at 50℃ for 0(as a control),10,15,20,25 and 30 min,respectively.The corrected mortality of T.castaneum was determined after 24 h.The results showed that different exposure times and heating rates had significant effects on the mortality of T.castaneum.The mortality of all life stages of T.castaneum with the same acclimation experience increased significantly with the increasing exposure time,and acclimation improved the heat tolerance of T.castaneum.The mortalities exposed at 50℃ for 30 min of T.castaneum eggs(acclimated to 46℃ at 5.00℃/h),larvae(acclimated to 44℃ at 2.50℃/h),pupae(acclimated to 46℃ at 5.00℃/h),and adults(acclimated to 46℃ at 2.50℃/h)were 62.50%,54.44%,58.89%and 53.33%,respectively,significantly lower than that of corresponding life stages of T.castaneum acclimated at 1.25 and 10.00℃/h.The present results are in favor of developing an effective heat treatment protocol for pest control.展开更多
Tribolium castaneum is a species of beetle in the family tenebrionidae,the darkling beetle.Conventional insecticides used for controling stored product pests are expensive and are proven to have side effects hence the...Tribolium castaneum is a species of beetle in the family tenebrionidae,the darkling beetle.Conventional insecticides used for controling stored product pests are expensive and are proven to have side effects hence the need to have an alternative biopesticides from plants which are proven to be effective against many insects.Azadrachta indica has been used for the control many insects,literature review show no report of its effects on Tribolium castaneum.This study is aimed at evaluating the bioinsecticidal activity of ethanolic leaves extract of Azadirachta indica against stored Tribolium castaneum.Different concentrations(60,70,80,90,and 100%)of ethanolic leaves extract of A.indica was applied on the filter paper and were allowed to dry.Control was done by applying water only on the filter paper.For the determination of the percentage mortality,15 adults of the insects was taken into the Petri dishes embedded with the filter paper covered with a lid.The A.indica extracts at different concentrations tested showed significant insecticidal activity against T.castaneum.Secondary metabolites screening indicated that the A.indica extract have secondary metabolites that are related to insecticidal activity.The study has shown that the ethanolic leaves extract A.indica is effective in managing stored product pest.展开更多
It is estimated that quarter to half a million specimens of the Nile crocodile (Crocodylus niloticus Laurenti, 1768) live in liberty. Ranching crocodiles for their skins has been successfully implemented in several Af...It is estimated that quarter to half a million specimens of the Nile crocodile (Crocodylus niloticus Laurenti, 1768) live in liberty. Ranching crocodiles for their skins has been successfully implemented in several African countries but also in Israel. Recently, in one of these ranches, an increased mortality of crocodile eggs was observed and it was thought that insects were causing their death. Two crocodile eggs were kept together with 100 adults of Tribolium castaneum (Herbst, 1797), while the third egg was kept without beetles. No food for beetles was added to the boxes. The eggs were observed twice a month for any kind of changes on their surface. After two months, at least 39 holes were observed in one egg and 146 on the second egg, which was exposed to beetles, while no holes could be seen in the control egg. Some of the holes were deep enough to see the inner layers of the egg. There is enough evidence to believe that adults of T. castaneum are able to damage the eggs of the Nile crocodile and in some cases lead to their death;measures should be taken to protect the eggs from this and similar species of beetles.展开更多
Evidence is accumulating that pattern recognition receptor(PRR)C-type lectins(CTL)play essential roles in recognition of pathogens.TcCTL14(accession no.TC00871)contains the most domains among all CTL of Tribolium cast...Evidence is accumulating that pattern recognition receptor(PRR)C-type lectins(CTL)play essential roles in recognition of pathogens.TcCTL14(accession no.TC00871)contains the most domains among all CTL of Tribolium castaneum.Yet the biological function of TcCTL14 remains unclear.In this study,TcCTL14 exhibiting typical motif and domain of CTL was cloned from T.castaneum.The expression pattern analysis showed that TcCTL14 was highly expressed in late pupae and central nervous system,and was upregulated after treatment with Escherichia coli and Staphylococcus aureus,respectively.Analysis of binding affinity revealed that recombinant TcCTL14 not only could bind to lipopolysaccharide and peptidoglycan in a dose-dependent fashion,but possibly could bind to and agglutinate different bacteria in a Ca^(2+)-dependent fashion.Knockdown of TcCTL14 before injection with bacteria led to the downregulation of nuclear factor-κB transcription factors of Toll/IMD and 4 antimicrobial peptides.Knockdown of TcCTL14 also caused suppressed metamorphosis,reduced fecundity,and delayed embryogenesis of T.castaneum.Further observation discovered that knockdown of TcCTL14 inhibited the development of ovaries and embryos.The detection of signaling pathways revealed that TcCTL14 may be involved in metamorphosis and fecundity by impacting 20-hydroxyecdysone and vitellogenin,respectively.Overall,these results indicate that TcCTL14 may contribute to immune response by agglutination or regulating the expression of antimicrobial peptides by the Toll/IMD pathway,and is required for T.castaneum development including metamorphosis,fecundity,and embryogenesis.These findings will improve the functional cognition of PRR CTL in insects and provide the new strategy for pest control.展开更多
Heat shock proteins (HSP) can protect organisms and cells from thermal damage. In this study, we cloned the full length cDNA encoding the HSP83 protein (the homologue of HSP90) of Tribolium castaneum(red flour be...Heat shock proteins (HSP) can protect organisms and cells from thermal damage. In this study, we cloned the full length cDNA encoding the HSP83 protein (the homologue of HSP90) of Tribolium castaneum(red flour beetle). The isolated cDNA contains the full coding sequence, a partial 5' untranslated region of 55 bp and the complete 3' untranslated region. We found the hsp83 gene is located on chromosome 5 of the T. castaneum genome. The predicted HSP83 protein sequence has a high similarity (on average 86.77%) with that of other insect species. The expression of the hsp83 gene in the whole body and in the ovary could be induced with heat stress (40℃ for 1 h) in newly hatched (within 3 h post emergence) and mature (10 days post emergence) beetles. Under normal conditions, the hsp83 expression in the ovary is about 3-fold higher than in the whole body at both stages. No significant difference in hsp83 expression was observed between the two ovarian developmental stages regardless if the beetles were treated with heat shock or not. The expression of the HSP83 protein in the whole body could also be induced with heat stress in newly hatched and mature beetles. However, in the ovary, HSP83 was only expressed in the follicle cells of mature beetles and not in newly hatched beetles, regardless if the beetles were treated with heat shock or not. Furthermore, the females were not able to produce mature oocytes after knock-down of the hsp83 expression by injecting dsRNA. These results suggest that the HSP83 protein is involved in protection against heat stress and could be involved in oogenesis during ovarian maturation of T. castaneum.展开更多
Long non-coding RNAs(lncRNAs)are poorly understood in insects.In this study,we performed genome-wide analysis of lncRNAs in Tribolium castaneum by RNA-seq.In total,4516 lncRNA transcripts corresponding to 3917 genes w...Long non-coding RNAs(lncRNAs)are poorly understood in insects.In this study,we performed genome-wide analysis of lncRNAs in Tribolium castaneum by RNA-seq.In total,4516 lncRNA transcripts corresponding to 3917 genes were identified from late embryos,early larvae,late larvae,early pupae,late pupae and early adults of T.castaneum,including 3152 novel lncRNAs and 1364 known lncRNAs.These lncRNAs have few exons and transcripts,and are short in length.During development,they exhibited nine different expression patterns.Functionally,they can act either by targeting messenger RNAs(1813 lncRNAs)and lncRNAs(45 lncRNAs)or as micro RNA(miRNA)precursors(46 lncRNAs).LncRNAs were observed to target the metabolic enzymes of glycolysis,TCA cycle and amino acids,demonstrating that lncRNAs control metabolism by regulating metabolic enzymes.Moreover,lncRNAs were shown to participate in cell differentiation and development via their targets.As miRNA precursors,lncRNAs could participate in the ecdysone signaling pathway.This study provides comprehensive information for lncRNAs of T.castaneum,and will promote functional analysis and target identification of lncRNAs in the insect.展开更多
Insects are a largely unexploited resource in prospecting for novel cellulolytic enzymes to improve the production of ethanol fuel from lignocellulosic biomass. The cost of lignocellulosic ethanol production is expect...Insects are a largely unexploited resource in prospecting for novel cellulolytic enzymes to improve the production of ethanol fuel from lignocellulosic biomass. The cost of lignocellulosic ethanol production is expected to decrease by the combination of cellulose degradation (saccharification) and fermentation of the resulting glucose to ethanol in a single process, catalyzed by the yeast Saccharomyces cerevisiae transformed to express efficient cellulases. While S. cerevisiae is an established heterologous expression system, there are no available data on the functional expression of insect cellulolytic enzymes for this species. To address this knowledge gap, S. cerevisiae was transformed to express the full-length cDNA encoding an endoglucanase from the red flour beetle, Tribolium castaneum (TcEG 1), and evaluated the activity of the transgenic product (rTcEG 1). Expression of the TcEG1 cDNA in S. cerevisiae was under control of the strong glyceraldehyde-3 phosphate dehydrogenase promoter. Cultured transformed yeast secreted rTcEG1 protein as a functional β-1,4-endoglucanase, which allowed transformants to survive on selective media containing cellulose as the only available carbon source. Evaluation of substrate specificity for secreted rTcEG1 demonstrated endoglucanase activity, although some activity was also detected against complex cellulose substrates. Potentially relevant to uses in biofuel production rTcEG1 activity increased with pH conditions, with the highest activity detected at pH 12. Our results demonstrate the potential for functional production of an insect cellulase in S. cerevisiae and confirm the stability of rTcEG1 activity in strong alkaline environments.展开更多
Chlorfenapyr, an insecticidal pyrrole, was applied to concrete arenas at concentrations of 1. 1, 0.825, 0.55, and 0.275 g of active ingredient [AI]/m^2. Adult Tribolium castaneum (Herbst), the red flour beetle, were...Chlorfenapyr, an insecticidal pyrrole, was applied to concrete arenas at concentrations of 1. 1, 0.825, 0.55, and 0.275 g of active ingredient [AI]/m^2. Adult Tribolium castaneum (Herbst), the red flour beetle, were exposed for 2, 4, or 8 h at each concentration, then removed and held either with or without food (wheat flour) for 7 days. Survival was assessed when the beetles were removed fi'om the exposure arenas and daily during the post- exposure period. In the presence of food, survival was high regardless of concentration and the day on which post-treatment survival was assessed, but survival did decrease as the exposure period increased from 4 to 8 h. When the beetles were not given food after exposure, survival at each concentration and exposure period declined during the 1-week post- exposure assessments. This pattern of decrease could be described by linear and non-linear equations. Results show the presence of food material greatly compromised effectiveness of the insecticide, and emphasize the importance of cleaning and sanitation in conjunction with insecticide treatments.展开更多
The sigma glutathione S-transferases (GSTSs) are a class of cytosolic glutathione S transferases (GSTs) that play important roles in antioxidant defense in insects, but the mechanisms by which GSTSs contribute to anti...The sigma glutathione S-transferases (GSTSs) are a class of cytosolic glutathione S transferases (GSTs) that play important roles in antioxidant defense in insects, but the mechanisms by which GSTSs contribute to antioxidant activity remain unclear. Here, we isolated a GSTS (GSTS6) from Tribolium castaneum and explored its function. Homology and phylogenetic analysis revealed that TcGSTS6 shared high identity with other evolutionarily conserved GSTSs. The recombinant TcGSTS6 protein had strong activity toward cumene hydroperoxide and 4-hydroxynonenal but low activity toward the universal substrate 1-chloro-2,4-dinitrobenzene. Exposure to various types of oxidative stress, including heat, cold, UV and pathogenic microbes, significantly induced TcGSTs6 expression, which indicates that it is involved in antioxidant defense. Knockdown TcGSTs6 by using RNA interference (RNAi) caused reduced antioxidant capacity, which was accomplished by cooperating with other antioxidant genes. Moreover, treatment with various insecticides such as phoxim, lambda-cyhalothrin, dichlorvos and carbofuran revealed that TcGSTS6 plays an important role in insecticide detoxification. The RNAi results showed that TcGSTS6 is essential for embryogenesis in T. castaneum. Our study elucidates the mechanism by which a GSTS contributes to antioxidant activity and enhances our understanding of the functional diversity of GSTSs in insects.展开更多
The zona pellucida domain protein Dusky(Dy)plays a vital role in wing morphogenesis in insects,but little information on its function has been reported.In this study,we found that dy regulated wing cell size,larval an...The zona pellucida domain protein Dusky(Dy)plays a vital role in wing morphogenesis in insects,but little information on its function has been reported.In this study,we found that dy regulated wing cell size,larval and pupal duration,and the metabolism of amino acid and 20-hydroxyecdysone in Tribolium castaneum.Using RNA-seq,413 differentially expressed genes were identified between physiological buffer-injected and dy-double-stranded RNA-treated larvae,including 88 downregulated genes and 325 upregulated genes.Among these genes,dy knockdown increased CYP18A1 expression to elevate the 26-hydroxylation of 20-hydroxyecdysone,which ultimately led to growth defects in wing cells.Silencing of dy upregulated the transcription of genes encoding tyrosine aminotransferase,4-hydroxyphenylpyruvate dioxygenase,homogentisate 1,2-dioxygenase,and Pale to promote the catabolism of tyrosine and phenylalanine,which eventually reduced amino acid content.Furthermore,dy knockdown upregulated 4E-BP expression,and 4E-BP silencing partially phenocopied dy RNA interference-mediated wing morphogenesis.These results suggest that Dy controls 20-hydroxyecdysone and amino acid metabolism to regulate wing morphogenesis in the insect.展开更多
The development of insect appendages requires the expression of multiple genes in a strict spatial and temporal order. The odd-skipped family genes are vital transcriptional factors involved in embryonic development. ...The development of insect appendages requires the expression of multiple genes in a strict spatial and temporal order. The odd-skipped family genes are vital transcriptional factors involved in embryonic development. The development and morphogenesis of the insect wing requires multiple transcription factors to regulate the expression of wing patterning genes at the transcriptional level. However, the function of odd-related genes in insect wing morphogenesis and development during postembryonic stages is unclear. We focused on the roles of the sister of odd and bowl (sob) gene, a member of odd-skipped family genes, during the wing morphopoiesis in Bombyx mori using the clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9 system and in Tribolium castaneum by RNA interference. The results showed that the wings were significantly smaller and degenerated, and wing veins were indistinct in the sob gene loss-of-function group in both B. mori and T. castaneum. Quantitative real-time polymerase chain reaction revealed that the Tcsob gene regulated the expression of wing development genes, such as the cht 7 and the vg gene. The findings suggest the importance of sob gene in insect wing morphology formation during postembryonic stages.展开更多
ABC transporters have been suggested to be involved in insecticide detoxification in different insect species mainly based on the indirect observation of transcriptional upregulation of ABC gene expression in response...ABC transporters have been suggested to be involved in insecticide detoxification in different insect species mainly based on the indirect observation of transcriptional upregulation of ABC gene expression in response to insecticide exposure.Previous studies performed by us and others in the red flour beetle,Tribolium castaneum,have analyzed the function of TcABCA-C and TcABCG-H genes using RNA interference(RNAi)and demonstrated that specific TcABCA and TcABCC genes are involved in the elimination of the pyrethroid tefluthrin and the benzoylurea diflubenzuron,because gene silencing increased the beetle's susceptibility to the insecticides.In this study,we focused on the potential functions of TcABCA-C genes in detoxification of the pyrethroid cyfluthrin(CF),the organophosphate malathion(MAL)and the diacylhdyazine tebufenozide(TBF).Analysis of transcript levels of selected TcABCA-C genes in response to treatment with these three chemically unrelated insecticides revealed that some genes were particularly upregulated after insecticide treatment.In addition,the ABC inhibitor verapamil synergized significantly the toxicity of MAL but only negligibly CF and TBF toxicities.Finally,silencing of two TcABCC genes by RNAi revealed a significant increase in susceptibility to MAL.In contrast,we did not observe a significant increase in insecticide-induced mortalities when knocking down TcABC genes in larvae treated with CF or TBF,although they were upregulated in response to insecticide treatment.Our results suggest that two pleiotropic ABCC transporters expressed in metabolic and excretory tissues contribute to the elimination of MAL.展开更多
文摘MicroRNAs (miRNAs) are endogenous 22-nt RNAs, which play important regulatory roles by post-transcriptional gene silencing. A computational strategy has been developed for the identification of conserved miRNAs based on features of known metazoan miRNAs in red flour beetle (Tribolium castaneum), which is regarded as one of the major laboratory models of arthropods. Among 118 putative miRNAs, 47% and 53% of the predicted miRNAs from the red flour beetle are harbored by known protein-coding genes (intronic) and genes located outside (intergenic miRNA), respectively. There are 31 intronic miRNAs in the same transcriptional orientation as the host genes, which may share RNA polymerase II and spliceosomal machinery with their host genes for their biogenesis. A hypothetical feed-back model has been proposed based on the analysis of the relationship between intronic miRNAs and their host genes in the development of red flour beetle.
基金funded in part by the National Natural Sciences Foundation of China (30871631)the Program for New Century Excellent Talents in University,China (NCET-04-0854)the Specialized Research Fund for the Doctoral Program of Higher Education of China(200806350009)
文摘An allele of CYP6BQI3, named CYP6BQ 13v2 (GenBank accession no. FJ209361), was isolated from the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) by RT-PCR. The cDNA sequence of CYP6BQ13v2, 1 563 bp in length, contains an open reading frame of 1 554 nucleotides encoding a putative protein of 518 amino acid residues with a predicted molecular weight of 59.92 kDa and a theoretical pl of 7.60. The putative protein contains the classic hemebinding sequence motif F××G×××C×G (residues 456-465) conserved among all P450 enzymes as well as other characteristic motifs of all cytochrome P450s. It shares 98% identity with the previously published sequence of CYP6BQ13 (GenBank accession no. XP967146) from the T. castaneum genome project. Phylogenetic analysis of amino acid sequences from members of various P450 families indicated that there was closer phylogenetic relationship of CYP6BQ 13v2 with CYP302A1 and CYP307A1 mediating synthesis of the insect molting hormone, distant relationship with CYP6B1 metabolizing plant allelochemicals, CYP6D 1 linking to pyrethroid resistance and other members of CYP6 family. Expression test of the gene in the adults and immature stages of T. castaneum by quantitative real-time PCR revealed that CYP6BQ13v2 is expressed in all life stages investigated. The mRNA expression level in 1st instar larvae was 14.9- and 3.86-fold higher than those in pupae and adults, respectively. The CYP6BQ13v2 expression levels appeared in the order of 1st instar larvae, followed by 4th instar larvae, 7th instar larvae, adult, and pupae from high to low. The more bioinformation of CYP6BQ 13v2 was also analyzed.
基金supported by the National Natural Science Foundation of China (31572326 and 31172146)the Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions, China
文摘The red flour beetle, Tribolium castaneum, is a major agriculture pest of stored grain, cereal products and peanuts for human consumption. It is reported that heat shock protein 18.3 of T. castaneum(Tchsp18.3) plays a significant role in stress resistance, development and reproduction. However, the regulatory systems of Tchsp18.3 remain unknown. Therefore, we compared the global transcriptome profiles of RNA interference(RNAi)-treated larvae(ds-Tchsp18.3) and control larvae of T. castaneum using RNA sequencing. Overall, we obtained 14 154 435 sequence reads aligned with 13 299 genes. Additionally, 569 differentially expressed genes(DEGs) were identified from the ds-Tchsp18.3 and control groups, of which 246 DEGs were annotated in the 47 Gene Ontology(GO) functional groups and 282 DEGs were assigned to 147 Kyoto Encyclopedia of Genes and Genomes(KEGG) biological signaling pathways. The DEGs encoding viperin, dorsal, Hdd11, PGRP2, defensin1 and defensin2 were simultaneously related to immunity and stress responses, which suggests that cross-talk might exist between the immunity and stress responses of T. castaneum. The knockdown of Tchsp18.3 gene expression suppressed the antioxidant activity process, which most likely modulated the effects of Tchsp18.3 on development and reproduction. Furthermore, the DEGs, including Blimp-1, Gld, Drm, Kinesin-14, Pthr2, Delta(11)-like and EGF-like domain protein 2, were also associated with the development and reproduction of ds-Tchsp18.3 insects. Additionally, knockdown of Tchsp18.3 amplified the serine protease(SP) signaling pathway to further regulate stress responses and innate immunity as well as development and reproduction of the red flour beetles. These results provide valuable insight into the molecular regulatory mechanism of Tchsp18.3 involved in insect physiology and further facilitate the research of suitable and sustainable management for pest control.
基金Supported by Special Fund for Technology Development of Research Institutes in Hainan Province(SQ2014KXSY0068)
文摘[Objeetive] The paper was to investigate the resistance of Tribolium castaneum to phosphine in Hainan Province. [Method] Using the measurement method of phosphine resistance of stored grain pests recommended by FAO, the phosphine resistance of eight strains of T. castaneum adults in Hainan Province was determined. [Result] The resistance coefficients of different strains of ?7. castaneum against phosphine ranged from 612 to 1 045, and all strains reached extremely high resistance level (Rf〉 160). When the concentration of phosphine was 200 mL/m^3, Rf612, Rf826.4 and Rfl 045 strains had complete lethal time of 8, 9 and 11 d, respectively. [Condusion] T. castarveum in Hainan Province had very strong resistance to phosphine, and the operation scheme of low concentration and long duration should be adopted in elimination of T. castaneum.
基金supported by National Natural Science Foundation of China(No.U1604115)Henan Provincial Key Scientific and Technological R&D Project(No.182102110111)。
文摘In order to elucidate the ability of stored product insects to resist high temperature at varying temperature conditions,the influence of different heating rates on the mortality of all life stages of Tribolium castaneum(Herbst)(Coleoptera:Tenebrionidae)was investigated.After acclimation from 28 to 42,44 and 46℃ at different heating rates(0,1.25,2.50,5.00,and 10.00℃/h),eggs,larvae,pupae,and adults of T.castaneum were exposed at 50℃ for 0(as a control),10,15,20,25 and 30 min,respectively.The corrected mortality of T.castaneum was determined after 24 h.The results showed that different exposure times and heating rates had significant effects on the mortality of T.castaneum.The mortality of all life stages of T.castaneum with the same acclimation experience increased significantly with the increasing exposure time,and acclimation improved the heat tolerance of T.castaneum.The mortalities exposed at 50℃ for 30 min of T.castaneum eggs(acclimated to 46℃ at 5.00℃/h),larvae(acclimated to 44℃ at 2.50℃/h),pupae(acclimated to 46℃ at 5.00℃/h),and adults(acclimated to 46℃ at 2.50℃/h)were 62.50%,54.44%,58.89%and 53.33%,respectively,significantly lower than that of corresponding life stages of T.castaneum acclimated at 1.25 and 10.00℃/h.The present results are in favor of developing an effective heat treatment protocol for pest control.
文摘Tribolium castaneum is a species of beetle in the family tenebrionidae,the darkling beetle.Conventional insecticides used for controling stored product pests are expensive and are proven to have side effects hence the need to have an alternative biopesticides from plants which are proven to be effective against many insects.Azadrachta indica has been used for the control many insects,literature review show no report of its effects on Tribolium castaneum.This study is aimed at evaluating the bioinsecticidal activity of ethanolic leaves extract of Azadirachta indica against stored Tribolium castaneum.Different concentrations(60,70,80,90,and 100%)of ethanolic leaves extract of A.indica was applied on the filter paper and were allowed to dry.Control was done by applying water only on the filter paper.For the determination of the percentage mortality,15 adults of the insects was taken into the Petri dishes embedded with the filter paper covered with a lid.The A.indica extracts at different concentrations tested showed significant insecticidal activity against T.castaneum.Secondary metabolites screening indicated that the A.indica extract have secondary metabolites that are related to insecticidal activity.The study has shown that the ethanolic leaves extract A.indica is effective in managing stored product pest.
文摘It is estimated that quarter to half a million specimens of the Nile crocodile (Crocodylus niloticus Laurenti, 1768) live in liberty. Ranching crocodiles for their skins has been successfully implemented in several African countries but also in Israel. Recently, in one of these ranches, an increased mortality of crocodile eggs was observed and it was thought that insects were causing their death. Two crocodile eggs were kept together with 100 adults of Tribolium castaneum (Herbst, 1797), while the third egg was kept without beetles. No food for beetles was added to the boxes. The eggs were observed twice a month for any kind of changes on their surface. After two months, at least 39 holes were observed in one egg and 146 on the second egg, which was exposed to beetles, while no holes could be seen in the control egg. Some of the holes were deep enough to see the inner layers of the egg. There is enough evidence to believe that adults of T. castaneum are able to damage the eggs of the Nile crocodile and in some cases lead to their death;measures should be taken to protect the eggs from this and similar species of beetles.
基金supported by grants from the National Natural Science Foundation of China to Bin Li(Grant number:31872970 and 32170521).
文摘Evidence is accumulating that pattern recognition receptor(PRR)C-type lectins(CTL)play essential roles in recognition of pathogens.TcCTL14(accession no.TC00871)contains the most domains among all CTL of Tribolium castaneum.Yet the biological function of TcCTL14 remains unclear.In this study,TcCTL14 exhibiting typical motif and domain of CTL was cloned from T.castaneum.The expression pattern analysis showed that TcCTL14 was highly expressed in late pupae and central nervous system,and was upregulated after treatment with Escherichia coli and Staphylococcus aureus,respectively.Analysis of binding affinity revealed that recombinant TcCTL14 not only could bind to lipopolysaccharide and peptidoglycan in a dose-dependent fashion,but possibly could bind to and agglutinate different bacteria in a Ca^(2+)-dependent fashion.Knockdown of TcCTL14 before injection with bacteria led to the downregulation of nuclear factor-κB transcription factors of Toll/IMD and 4 antimicrobial peptides.Knockdown of TcCTL14 also caused suppressed metamorphosis,reduced fecundity,and delayed embryogenesis of T.castaneum.Further observation discovered that knockdown of TcCTL14 inhibited the development of ovaries and embryos.The detection of signaling pathways revealed that TcCTL14 may be involved in metamorphosis and fecundity by impacting 20-hydroxyecdysone and vitellogenin,respectively.Overall,these results indicate that TcCTL14 may contribute to immune response by agglutination or regulating the expression of antimicrobial peptides by the Toll/IMD pathway,and is required for T.castaneum development including metamorphosis,fecundity,and embryogenesis.These findings will improve the functional cognition of PRR CTL in insects and provide the new strategy for pest control.
基金supported by the National Basic Research Program of China (No. 2006CB102104)the Key Project of the National Natural Science Foundation of China (No. 30430500)
文摘Heat shock proteins (HSP) can protect organisms and cells from thermal damage. In this study, we cloned the full length cDNA encoding the HSP83 protein (the homologue of HSP90) of Tribolium castaneum(red flour beetle). The isolated cDNA contains the full coding sequence, a partial 5' untranslated region of 55 bp and the complete 3' untranslated region. We found the hsp83 gene is located on chromosome 5 of the T. castaneum genome. The predicted HSP83 protein sequence has a high similarity (on average 86.77%) with that of other insect species. The expression of the hsp83 gene in the whole body and in the ovary could be induced with heat stress (40℃ for 1 h) in newly hatched (within 3 h post emergence) and mature (10 days post emergence) beetles. Under normal conditions, the hsp83 expression in the ovary is about 3-fold higher than in the whole body at both stages. No significant difference in hsp83 expression was observed between the two ovarian developmental stages regardless if the beetles were treated with heat shock or not. The expression of the HSP83 protein in the whole body could also be induced with heat stress in newly hatched and mature beetles. However, in the ovary, HSP83 was only expressed in the follicle cells of mature beetles and not in newly hatched beetles, regardless if the beetles were treated with heat shock or not. Furthermore, the females were not able to produce mature oocytes after knock-down of the hsp83 expression by injecting dsRNA. These results suggest that the HSP83 protein is involved in protection against heat stress and could be involved in oogenesis during ovarian maturation of T. castaneum.
基金National Natural Science Foundation of China(Nos.31601893,31572326 and 31572467)Young Talents Training Program of Jiangsu University,and Postgraduate Research&Practice Inno-vation Program of Jiangsu Province(KYCX19_1576).
文摘Long non-coding RNAs(lncRNAs)are poorly understood in insects.In this study,we performed genome-wide analysis of lncRNAs in Tribolium castaneum by RNA-seq.In total,4516 lncRNA transcripts corresponding to 3917 genes were identified from late embryos,early larvae,late larvae,early pupae,late pupae and early adults of T.castaneum,including 3152 novel lncRNAs and 1364 known lncRNAs.These lncRNAs have few exons and transcripts,and are short in length.During development,they exhibited nine different expression patterns.Functionally,they can act either by targeting messenger RNAs(1813 lncRNAs)and lncRNAs(45 lncRNAs)or as micro RNA(miRNA)precursors(46 lncRNAs).LncRNAs were observed to target the metabolic enzymes of glycolysis,TCA cycle and amino acids,demonstrating that lncRNAs control metabolism by regulating metabolic enzymes.Moreover,lncRNAs were shown to participate in cell differentiation and development via their targets.As miRNA precursors,lncRNAs could participate in the ecdysone signaling pathway.This study provides comprehensive information for lncRNAs of T.castaneum,and will promote functional analysis and target identification of lncRNAs in the insect.
文摘Insects are a largely unexploited resource in prospecting for novel cellulolytic enzymes to improve the production of ethanol fuel from lignocellulosic biomass. The cost of lignocellulosic ethanol production is expected to decrease by the combination of cellulose degradation (saccharification) and fermentation of the resulting glucose to ethanol in a single process, catalyzed by the yeast Saccharomyces cerevisiae transformed to express efficient cellulases. While S. cerevisiae is an established heterologous expression system, there are no available data on the functional expression of insect cellulolytic enzymes for this species. To address this knowledge gap, S. cerevisiae was transformed to express the full-length cDNA encoding an endoglucanase from the red flour beetle, Tribolium castaneum (TcEG 1), and evaluated the activity of the transgenic product (rTcEG 1). Expression of the TcEG1 cDNA in S. cerevisiae was under control of the strong glyceraldehyde-3 phosphate dehydrogenase promoter. Cultured transformed yeast secreted rTcEG1 protein as a functional β-1,4-endoglucanase, which allowed transformants to survive on selective media containing cellulose as the only available carbon source. Evaluation of substrate specificity for secreted rTcEG1 demonstrated endoglucanase activity, although some activity was also detected against complex cellulose substrates. Potentially relevant to uses in biofuel production rTcEG1 activity increased with pH conditions, with the highest activity detected at pH 12. Our results demonstrate the potential for functional production of an insect cellulase in S. cerevisiae and confirm the stability of rTcEG1 activity in strong alkaline environments.
文摘Chlorfenapyr, an insecticidal pyrrole, was applied to concrete arenas at concentrations of 1. 1, 0.825, 0.55, and 0.275 g of active ingredient [AI]/m^2. Adult Tribolium castaneum (Herbst), the red flour beetle, were exposed for 2, 4, or 8 h at each concentration, then removed and held either with or without food (wheat flour) for 7 days. Survival was assessed when the beetles were removed fi'om the exposure arenas and daily during the post- exposure period. In the presence of food, survival was high regardless of concentration and the day on which post-treatment survival was assessed, but survival did decrease as the exposure period increased from 4 to 8 h. When the beetles were not given food after exposure, survival at each concentration and exposure period declined during the 1-week post- exposure assessments. This pattern of decrease could be described by linear and non-linear equations. Results show the presence of food material greatly compromised effectiveness of the insecticide, and emphasize the importance of cleaning and sanitation in conjunction with insecticide treatments.
基金This work was supported by the National Natural Science Foundation of China(No.31872970&31572326).
文摘The sigma glutathione S-transferases (GSTSs) are a class of cytosolic glutathione S transferases (GSTs) that play important roles in antioxidant defense in insects, but the mechanisms by which GSTSs contribute to antioxidant activity remain unclear. Here, we isolated a GSTS (GSTS6) from Tribolium castaneum and explored its function. Homology and phylogenetic analysis revealed that TcGSTS6 shared high identity with other evolutionarily conserved GSTSs. The recombinant TcGSTS6 protein had strong activity toward cumene hydroperoxide and 4-hydroxynonenal but low activity toward the universal substrate 1-chloro-2,4-dinitrobenzene. Exposure to various types of oxidative stress, including heat, cold, UV and pathogenic microbes, significantly induced TcGSTs6 expression, which indicates that it is involved in antioxidant defense. Knockdown TcGSTs6 by using RNA interference (RNAi) caused reduced antioxidant capacity, which was accomplished by cooperating with other antioxidant genes. Moreover, treatment with various insecticides such as phoxim, lambda-cyhalothrin, dichlorvos and carbofuran revealed that TcGSTS6 plays an important role in insecticide detoxification. The RNAi results showed that TcGSTS6 is essential for embryogenesis in T. castaneum. Our study elucidates the mechanism by which a GSTS contributes to antioxidant activity and enhances our understanding of the functional diversity of GSTSs in insects.
基金supported by the National Natural Science Foundation of China(Nos.32070501 and 31601893)General Project of the Natural Science Foundation of the Jiangsu Higher Education Institu・tions(No.16KJB180004)Young Talents Training Pto・gram of Jiangsu University,and Postgraduate Research&Practice Innovation Program of Jiangsu Province(KYCX19_1576).
文摘The zona pellucida domain protein Dusky(Dy)plays a vital role in wing morphogenesis in insects,but little information on its function has been reported.In this study,we found that dy regulated wing cell size,larval and pupal duration,and the metabolism of amino acid and 20-hydroxyecdysone in Tribolium castaneum.Using RNA-seq,413 differentially expressed genes were identified between physiological buffer-injected and dy-double-stranded RNA-treated larvae,including 88 downregulated genes and 325 upregulated genes.Among these genes,dy knockdown increased CYP18A1 expression to elevate the 26-hydroxylation of 20-hydroxyecdysone,which ultimately led to growth defects in wing cells.Silencing of dy upregulated the transcription of genes encoding tyrosine aminotransferase,4-hydroxyphenylpyruvate dioxygenase,homogentisate 1,2-dioxygenase,and Pale to promote the catabolism of tyrosine and phenylalanine,which eventually reduced amino acid content.Furthermore,dy knockdown upregulated 4E-BP expression,and 4E-BP silencing partially phenocopied dy RNA interference-mediated wing morphogenesis.These results suggest that Dy controls 20-hydroxyecdysone and amino acid metabolism to regulate wing morphogenesis in the insect.
基金This project received funding from the National Natural Science Foundation of China(No.U20A2058,No.31830094,No.31472153)Funds of China Agriculture Research System(No.CARS-18-ZJO102).
文摘The development of insect appendages requires the expression of multiple genes in a strict spatial and temporal order. The odd-skipped family genes are vital transcriptional factors involved in embryonic development. The development and morphogenesis of the insect wing requires multiple transcription factors to regulate the expression of wing patterning genes at the transcriptional level. However, the function of odd-related genes in insect wing morphogenesis and development during postembryonic stages is unclear. We focused on the roles of the sister of odd and bowl (sob) gene, a member of odd-skipped family genes, during the wing morphopoiesis in Bombyx mori using the clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9 system and in Tribolium castaneum by RNA interference. The results showed that the wings were significantly smaller and degenerated, and wing veins were indistinct in the sob gene loss-of-function group in both B. mori and T. castaneum. Quantitative real-time polymerase chain reaction revealed that the Tcsob gene regulated the expression of wing development genes, such as the cht 7 and the vg gene. The findings suggest the importance of sob gene in insect wing morphology formation during postembryonic stages.
基金We are grateful to Johanne Tietmeyer for generating the used pGEM-T plasmids containing TcABC-cDNA inserts.This work was supported by a grant from the Deutsche Forschungsgemeinschaft(DFG Me2210/4-1).
文摘ABC transporters have been suggested to be involved in insecticide detoxification in different insect species mainly based on the indirect observation of transcriptional upregulation of ABC gene expression in response to insecticide exposure.Previous studies performed by us and others in the red flour beetle,Tribolium castaneum,have analyzed the function of TcABCA-C and TcABCG-H genes using RNA interference(RNAi)and demonstrated that specific TcABCA and TcABCC genes are involved in the elimination of the pyrethroid tefluthrin and the benzoylurea diflubenzuron,because gene silencing increased the beetle's susceptibility to the insecticides.In this study,we focused on the potential functions of TcABCA-C genes in detoxification of the pyrethroid cyfluthrin(CF),the organophosphate malathion(MAL)and the diacylhdyazine tebufenozide(TBF).Analysis of transcript levels of selected TcABCA-C genes in response to treatment with these three chemically unrelated insecticides revealed that some genes were particularly upregulated after insecticide treatment.In addition,the ABC inhibitor verapamil synergized significantly the toxicity of MAL but only negligibly CF and TBF toxicities.Finally,silencing of two TcABCC genes by RNAi revealed a significant increase in susceptibility to MAL.In contrast,we did not observe a significant increase in insecticide-induced mortalities when knocking down TcABC genes in larvae treated with CF or TBF,although they were upregulated in response to insecticide treatment.Our results suggest that two pleiotropic ABCC transporters expressed in metabolic and excretory tissues contribute to the elimination of MAL.