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
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
Insect symbionts Wolbachia used for pest population control has focused on vector pest species and agricultural insects while rare reports in Chinese stored-product insect samples.In this paper,we surveyed the prevale...Insect symbionts Wolbachia used for pest population control has focused on vector pest species and agricultural insects while rare reports in Chinese stored-product insect samples.In this paper,we surveyed the prevalence of Wolbachia using a PCR detection method in ten Tenebrionidae stored-product insects.Subsequently,the spatiotemporal Wolbachia infection dynamics in Tribolium confusum and Wolbachia elimination patterns using tetracycline treatment were investigated in detail by TaqMan®probe real-time quantitative PCR,and host reproductive fitness parameters were compared.T.confusum was the only Wolbachia infected species in all the surveyed species.Wolbachia infection density consistently increased with the development of T.confusumand plateaued at 3.7×107 wsp copies per individual insect at the young adult stage.Wolbachia densities in females were higher than that in males with a significant difference at the pupae stage and varied among different tissues and organs.Uninfected female beetles were completely incapable of producing mature progenies when crossed with Wolbachia infected males.Embryogenesis and egg hatch rate were specifically inhibited after Wolbachia elimination,while other traits,including the number of eggs,pupation rate and sex ratio,remained unaffected by tetracycline treatment.Our results showthat the TaqMan®probe qPCR is a reliable detection method for quantifying the density of Wolbachia as compared to qualitative detection of wsp gene by PCR and relatively quantified by real-time qPCR.The fitness results indicated that Wolbachia infection was not an obligate symbiont and benefited the host confused flour beetle.展开更多
Seven biscuit types namely, Okin, Digestive, Cabin, Peanut, Cream crackers, Hobnobs and Glucose and wheat flour were screened for their resistance to Tribolium castaneum (Herbst) at ambient temperature of 28±2 ℃...Seven biscuit types namely, Okin, Digestive, Cabin, Peanut, Cream crackers, Hobnobs and Glucose and wheat flour were screened for their resistance to Tribolium castaneum (Herbst) at ambient temperature of 28±2 ℃ and 78±2% relative humidity in the laboratory. 50 g of each biscuit sample were infested with four female and two male adult beetles and left for 15 days then remove for a resistant experiment and 70 days for a mortality experiment. Each treatment and the control without beetles were replicated three times. Results showed that there was significant difference (P<0.05) in the mortality rate of T. castaneum in the biscuit types and wheat flour samples, while there was no significant difference (P>0.05) in the weight loss of biscuit types. The highest adult mortality of beetles (100%) was obtained from Peanut biscuits at 28 days after infestation while there was only 5.5% adult mortality in wheat flour at 70 days after infestation. The susceptibility index was 0 for all the biscuit types since there was no adult emergence indicating that they were resistant to T. castaneum infestation. Resistance in the biscuit types could be due to chemical additives (e.g. sodium chloride and sodium bicarbonate) used in the production of biscuits, which may have inhibitory effect on the development of T. castaneum and also the low moisture content of the biscuits. Packages that can easily be perforated or damaged to allow absorption of moisture from the environment should not be used in packing biscuits.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 this study, a rapid molecular identification method of Tribolium destructor was established with PCR and PCR-RFLP （polymerase chain reaction-restriction fragment length polymorphism） technology. According to the ...In this study, a rapid molecular identification method of Tribolium destructor was established with PCR and PCR-RFLP （polymerase chain reaction-restriction fragment length polymorphism） technology. According to the results, （ 1 ） with PCR method, specific primers were designed based on CO1 gene of T. destructor for PCR amplification, and electrophoresis detection confirmed that PCR method could be used to rapidly and accurately identify T. destructor; （2） with PCR-RFLP method, two pairs of degenerate primers were used to amplify CO1 gene of Tribolium species, PCR products were digested with HindIII and detected by electrophoresis, results indicated that PCR-RFLP method could also be used for rapid identification of T. destructor in quarantine practice.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
Ethanolic extracts of five plant material from Eucalyptus gluaca, Melia azadrach, Mentha arvensis, Olea europaea leaves and periearp of Punica granatum were investigated for insecticidal activity against the confused ...Ethanolic extracts of five plant material from Eucalyptus gluaca, Melia azadrach, Mentha arvensis, Olea europaea leaves and periearp of Punica granatum were investigated for insecticidal activity against the confused flour beetle Tribolium confusum DuVal in the laboratory. Different concentrations 2.5%, 5%, 7.5%, 10% using filter paper test were implemented in the experiment. Mortality was assessed after 24, 48, 72 hrs of treatment and LDs0 were calculated at respective time intervals. Data analysis was made by using factorial Complete Randomized Design （CRD）. The results revealed that all of the tested materials with some variations had lethal effects against the pest as compared with the untreated check. The time 24 h showed higher insecticidal effect with the tested concentrations for the larvae and adult of Tribolium confusum. Strong toxicity effect was observed at concentration 5% of M.. arvensis for adult followed by E. gluaca, O. europaea, M.. azedrach and P. granatum. For larval stage the extract of O. europaea and E. gluaca with concentration 7.5%, 1%, respectively caused high mortality followed by M. arvensis, M. azedrach and P. granatum. The LDs0 values for adults and larvae were calculated to be 1.33 and 1.42 with M.. arvensis and O. europaea, respectively.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
Hemocytes are pivotal in the immune response of insects against invasive pathogens.However,our knowledge of hemocyte types and their specific function in Tribolium castaneum,an increasingly important Coleoptera model ...Hemocytes are pivotal in the immune response of insects against invasive pathogens.However,our knowledge of hemocyte types and their specific function in Tribolium castaneum,an increasingly important Coleoptera model insect in various research fields,remains limited.Presently,a combination of morphological criteria and dye-staining properties were used to characterize hemocyte types from T.castaneum larvae,and 4 distinct types were identified:granulocytes,oenocytoids,plasmatocytes and prohemocytes.Following different immune challenges,the total hemocyte counts declined rapidly in the initial phase(at 2 h),then increased over time(at 4 and 6 h)and eventually returned to the naive state by 24 h post-injection.Notably,the morphology of granulocytes underwent dramatic changes,characterized by an expansion of the surface area and an increased production of pseudopods,and with the number of granulocytes rising significantly through mitotic division.Granulocytes and plasmatocytes,the main hemocyte types in T.castaneum larvae,can phagocytose bacteria or latex beads injected into the larval hemolymph in vivo.Furthermore,these hemocytes participate in the encapsulation and melanization processes in vitro,forming capsules to encapsulate and melanize nickel-nitrilotriacetic acid(Ni-NTA)beads.This study provides the first comprehensive characterization of circulating hemocytes in T.castaneum larvae,offering valuable insights into cell-mediated immunity in response to bacterial infection and the injection of latex beads.These results deepen our understanding of the cellular response mechanisms in T.castaneum larvae and lay a solid foundation for subsequent investigations of the involvement of T.castaneum hemocytes in combating pathogens.展开更多
The clustered regularly interspaced small palindromic repeats(CRISPR)/CRISPR-associated nuclease 9(Cas9)-mediated gene editing technology has revolutionized the study of fundamental biological questions in various ins...The clustered regularly interspaced small palindromic repeats(CRISPR)/CRISPR-associated nuclease 9(Cas9)-mediated gene editing technology has revolutionized the study of fundamental biological questions in various insects.Diverse approaches have been developed to deliver the single-guide RNA(sgRNA)and Cas9 to the nucleus of insect embryos or oocytes to achieve gene editing,including the predominant embryonic injection methods and alternative protocols through parental ovary delivery.However,a systematic comparative study of these approaches is limited,especially within a given insect.Here,we focused on revealing the detailed differences in CRISPR/Cas9-mediated gene editing between the embryo and ovary delivery methods in the beetle Tribolium castaneum,using the cardinal and tyrosine hydroxylase(TH)as reporter genes.We demonstrated that both genes could be efficiently edited by delivering Cas9/sgRNA ribonucleoproteins to the embryos by microinjection,leading to the mutant phenotypes and indels in the target gene sites.Next,the Cas9/sgRNA complex,coupled with a nanocarrier called Branched Amphiphilic Peptide Capsules(BAPC),were delivered to the ovaries of parental females to examine the efficacy of BAPC-mediated gene editing.Although we observed that a small number of beetles’progeny targeting the cardinal exhibited the expected white-eye phenotype,unexpectedly,no target DNA indels were found following subsequent sequencing analysis.In addition,we adopted a novel approach termed“direct parental”CRISPR(DIPA-CRISPR).However,we still failed to find gene-editing events in the cardinal or TH gene-targeted insects.Our results indicate that the conventional embryonic injection of CRISPR is an effective method to initiate genome editing in T.castaneum.However,it is inefficient by the parental ovary delivery approach.展开更多
Small heat shock proteins (sHSPs) are diverse and mainly function as molecular chaperones to protect organisms and cells from various stresses. In this study, hsp 18.3, one Tribolium castaneum species-specific shsp、h...Small heat shock proteins (sHSPs) are diverse and mainly function as molecular chaperones to protect organisms and cells from various stresses. In this study, hsp 18.3, one Tribolium castaneum species-specific shsp、has been identified. Quantitative real-time polymerase chain reaction illustrated that Tchsp 18.3 is expressed in all developmental stages, and is highly expressed at early pupal and late adult stages, while it is highly expressed in ovary and fat body at the adult period. Moreover, it was up-regulated 4532 ± 396-fold in response to enhanced heat stress but not to cold stress;meanwhile the lifespan of adults in ds-Tchsp 18.3 group reduced by 15.8% from control group under starvation. Laval RNA interference (RNAi) of Tchsp 18.3 caused 86.1%±4.5% arrested pupal eclosion and revealed that Tchsp 18.3 played an important role in insect development. In addition, parental RNAi of Tchsp 18.3 reduced the oviposition amount by 94.7%. These results suggest that Tchsp 18.3 is not only essential for the resistance to heat and starvation stress, but also is critical for normal development and reproduction in T. castaneum.展开更多
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.
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 National Natural Science Fund Project (No.31601890)the Fundamental Research Funds for the Henan Provincial Colleges and Universities in Henan University of Technology (No.2016XTCX01)
文摘Insect symbionts Wolbachia used for pest population control has focused on vector pest species and agricultural insects while rare reports in Chinese stored-product insect samples.In this paper,we surveyed the prevalence of Wolbachia using a PCR detection method in ten Tenebrionidae stored-product insects.Subsequently,the spatiotemporal Wolbachia infection dynamics in Tribolium confusum and Wolbachia elimination patterns using tetracycline treatment were investigated in detail by TaqMan®probe real-time quantitative PCR,and host reproductive fitness parameters were compared.T.confusum was the only Wolbachia infected species in all the surveyed species.Wolbachia infection density consistently increased with the development of T.confusumand plateaued at 3.7×107 wsp copies per individual insect at the young adult stage.Wolbachia densities in females were higher than that in males with a significant difference at the pupae stage and varied among different tissues and organs.Uninfected female beetles were completely incapable of producing mature progenies when crossed with Wolbachia infected males.Embryogenesis and egg hatch rate were specifically inhibited after Wolbachia elimination,while other traits,including the number of eggs,pupation rate and sex ratio,remained unaffected by tetracycline treatment.Our results showthat the TaqMan®probe qPCR is a reliable detection method for quantifying the density of Wolbachia as compared to qualitative detection of wsp gene by PCR and relatively quantified by real-time qPCR.The fitness results indicated that Wolbachia infection was not an obligate symbiont and benefited the host confused flour beetle.
文摘Seven biscuit types namely, Okin, Digestive, Cabin, Peanut, Cream crackers, Hobnobs and Glucose and wheat flour were screened for their resistance to Tribolium castaneum (Herbst) at ambient temperature of 28±2 ℃ and 78±2% relative humidity in the laboratory. 50 g of each biscuit sample were infested with four female and two male adult beetles and left for 15 days then remove for a resistant experiment and 70 days for a mortality experiment. Each treatment and the control without beetles were replicated three times. Results showed that there was significant difference (P<0.05) in the mortality rate of T. castaneum in the biscuit types and wheat flour samples, while there was no significant difference (P>0.05) in the weight loss of biscuit types. The highest adult mortality of beetles (100%) was obtained from Peanut biscuits at 28 days after infestation while there was only 5.5% adult mortality in wheat flour at 70 days after infestation. The susceptibility index was 0 for all the biscuit types since there was no adult emergence indicating that they were resistant to T. castaneum infestation. Resistance in the biscuit types could be due to chemical additives (e.g. sodium chloride and sodium bicarbonate) used in the production of biscuits, which may have inhibitory effect on the development of T. castaneum and also the low moisture content of the biscuits. Packages that can easily be perforated or damaged to allow absorption of moisture from the environment should not be used in packing biscuits.
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 Science and Technology Project of Jiangsu Entry and Exit Inspection and Quarantine Bureau(2010KJ06)
文摘In this study, a rapid molecular identification method of Tribolium destructor was established with PCR and PCR-RFLP （polymerase chain reaction-restriction fragment length polymorphism） technology. According to the results, （ 1 ） with PCR method, specific primers were designed based on CO1 gene of T. destructor for PCR amplification, and electrophoresis detection confirmed that PCR method could be used to rapidly and accurately identify T. destructor; （2） with PCR-RFLP method, two pairs of degenerate primers were used to amplify CO1 gene of Tribolium species, PCR products were digested with HindIII and detected by electrophoresis, results indicated that PCR-RFLP method could also be used for rapid identification of T. destructor in quarantine practice.
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.
文摘Ethanolic extracts of five plant material from Eucalyptus gluaca, Melia azadrach, Mentha arvensis, Olea europaea leaves and periearp of Punica granatum were investigated for insecticidal activity against the confused flour beetle Tribolium confusum DuVal in the laboratory. Different concentrations 2.5%, 5%, 7.5%, 10% using filter paper test were implemented in the experiment. Mortality was assessed after 24, 48, 72 hrs of treatment and LDs0 were calculated at respective time intervals. Data analysis was made by using factorial Complete Randomized Design （CRD）. The results revealed that all of the tested materials with some variations had lethal effects against the pest as compared with the untreated check. The time 24 h showed higher insecticidal effect with the tested concentrations for the larvae and adult of Tribolium confusum. Strong toxicity effect was observed at concentration 5% of M.. arvensis for adult followed by E. gluaca, O. europaea, M.. azedrach and P. granatum. For larval stage the extract of O. europaea and E. gluaca with concentration 7.5%, 1%, respectively caused high mortality followed by M. arvensis, M. azedrach and P. granatum. The LDs0 values for adults and larvae were calculated to be 1.33 and 1.42 with M.. arvensis and O. europaea, respectively.
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.
基金supported by grants from the National Natural Science Foundation of China(32170521)Jiangsu Province Higher Education Basic Science(Natural Science)Research Project(23KJD210002)。
文摘Hemocytes are pivotal in the immune response of insects against invasive pathogens.However,our knowledge of hemocyte types and their specific function in Tribolium castaneum,an increasingly important Coleoptera model insect in various research fields,remains limited.Presently,a combination of morphological criteria and dye-staining properties were used to characterize hemocyte types from T.castaneum larvae,and 4 distinct types were identified:granulocytes,oenocytoids,plasmatocytes and prohemocytes.Following different immune challenges,the total hemocyte counts declined rapidly in the initial phase(at 2 h),then increased over time(at 4 and 6 h)and eventually returned to the naive state by 24 h post-injection.Notably,the morphology of granulocytes underwent dramatic changes,characterized by an expansion of the surface area and an increased production of pseudopods,and with the number of granulocytes rising significantly through mitotic division.Granulocytes and plasmatocytes,the main hemocyte types in T.castaneum larvae,can phagocytose bacteria or latex beads injected into the larval hemolymph in vivo.Furthermore,these hemocytes participate in the encapsulation and melanization processes in vitro,forming capsules to encapsulate and melanize nickel-nitrilotriacetic acid(Ni-NTA)beads.This study provides the first comprehensive characterization of circulating hemocytes in T.castaneum larvae,offering valuable insights into cell-mediated immunity in response to bacterial infection and the injection of latex beads.These results deepen our understanding of the cellular response mechanisms in T.castaneum larvae and lay a solid foundation for subsequent investigations of the involvement of T.castaneum hemocytes in combating pathogens.
基金funded by the National Key R&D Program of China(2017YFD0200900)the National Natural Science Foundation of China(31101432).
文摘The clustered regularly interspaced small palindromic repeats(CRISPR)/CRISPR-associated nuclease 9(Cas9)-mediated gene editing technology has revolutionized the study of fundamental biological questions in various insects.Diverse approaches have been developed to deliver the single-guide RNA(sgRNA)and Cas9 to the nucleus of insect embryos or oocytes to achieve gene editing,including the predominant embryonic injection methods and alternative protocols through parental ovary delivery.However,a systematic comparative study of these approaches is limited,especially within a given insect.Here,we focused on revealing the detailed differences in CRISPR/Cas9-mediated gene editing between the embryo and ovary delivery methods in the beetle Tribolium castaneum,using the cardinal and tyrosine hydroxylase(TH)as reporter genes.We demonstrated that both genes could be efficiently edited by delivering Cas9/sgRNA ribonucleoproteins to the embryos by microinjection,leading to the mutant phenotypes and indels in the target gene sites.Next,the Cas9/sgRNA complex,coupled with a nanocarrier called Branched Amphiphilic Peptide Capsules(BAPC),were delivered to the ovaries of parental females to examine the efficacy of BAPC-mediated gene editing.Although we observed that a small number of beetles’progeny targeting the cardinal exhibited the expected white-eye phenotype,unexpectedly,no target DNA indels were found following subsequent sequencing analysis.In addition,we adopted a novel approach termed“direct parental”CRISPR(DIPA-CRISPR).However,we still failed to find gene-editing events in the cardinal or TH gene-targeted insects.Our results indicate that the conventional embryonic injection of CRISPR is an effective method to initiate genome editing in T.castaneum.However,it is inefficient by the parental ovary delivery approach.
基金the National Natural Science Foundation of China (No. 31572326 & 31172146)the Natural Science Foundation of Jiangsu Province, China (No. BK2011785)the PAPD of Jiangsu Higher Education Institutions.
文摘Small heat shock proteins (sHSPs) are diverse and mainly function as molecular chaperones to protect organisms and cells from various stresses. In this study, hsp 18.3, one Tribolium castaneum species-specific shsp、has been identified. Quantitative real-time polymerase chain reaction illustrated that Tchsp 18.3 is expressed in all developmental stages, and is highly expressed at early pupal and late adult stages, while it is highly expressed in ovary and fat body at the adult period. Moreover, it was up-regulated 4532 ± 396-fold in response to enhanced heat stress but not to cold stress;meanwhile the lifespan of adults in ds-Tchsp 18.3 group reduced by 15.8% from control group under starvation. Laval RNA interference (RNAi) of Tchsp 18.3 caused 86.1%±4.5% arrested pupal eclosion and revealed that Tchsp 18.3 played an important role in insect development. In addition, parental RNAi of Tchsp 18.3 reduced the oviposition amount by 94.7%. These results suggest that Tchsp 18.3 is not only essential for the resistance to heat and starvation stress, but also is critical for normal development and reproduction in T. castaneum.
