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Sample records for bombyx

  1. Bombyx mori

    Indian Academy of Sciences (India)

    2010-09-06

    Sep 6, 2010 ... 2008 Identification of the female-determining region of the W chromosome in Bombyx mori. Genetica 133, 269–282. Abe H., Fujii T. and Shimada T. 2010 Sex chromosomes and sex determination in Bombyx mori. In Molecular biology and genet- ics of the Lepidoptera (ed. M. R. Goldsmith and F. Marec), pp.

  2. Bombyx mori L.; Bombycidae

    African Journals Online (AJOL)

    AJL

    2011-09-28

    Sep 28, 2011 ... excellent results of egg hatchability, pupation and survival rate (low larval mortality). ... affecting the quality and quantity of cocoon crops. ..... Health, 6(3-4): 3-5. Ramesh C, Seshagiri SV, Rao CGP (2009). Evaluation and identification of superior polyvoltine crossbreeds of mulberry silk worm, Bombyx mori ...

  3. Protective Effect of Bombyx mori L Cocoon (Abresham) and its ...

    African Journals Online (AJOL)

    HP

    Bombyx mori (Lepidoptera: Bombycidae), is one of the most important insects feed on the leaves of the mulberry family Moraceae, genus Morus. [6]. The cocoon shell of the silkworm Bombyx mori consists of silk fibroin fiber (70 %) surrounded by a sericin layer made up of sericin (25 %) and non-sericin (5 %) components.

  4. Resistance comparison of domesticated silkworm (Bombyx mori L ...

    African Journals Online (AJOL)

    USER

    2010-03-22

    Mar 22, 2010 ... Yukuhiro K, Sezutsu H, Itoh M, Shimizu K, Banno Y (2002). Significant levels of sequence divergence and gene rearrangements have occurred between the mitochondrial genomes of the wild mulberry silkmoth, Bombyx mandarina, and its close relative, the domesticated. Silkmoth, Bombyx mori. Mol. Biol.

  5. Molecular phylogeny of the domesticated silkworm, Bombyx mori ...

    Indian Academy of Sciences (India)

    Unknown

    stranded DNA molecule, which is known to have small molecular weight, rapid nucleotide ... Keywords. Bombyx mori; Bombyx mandarina; mitochondrial DNA; cytochrome b gene; phylogenetic tree. *For correspondence. E-mail: .... By sequencing the PCR products, the 12 5′-end seq- uences of CytB (591 bp) were ...

  6. RNA recognition motif (RRM)-containing proteins in Bombyx mori

    African Journals Online (AJOL)

    STORAGESEVER

    2009-03-20

    ' TBPH, suggesting that it may play some roles in emergency responses. Key words: Bombyx mori, RRM-containing protein, TAR DNA-binding protein-43. INTRODUCTION. RNA binding proteins constitute an extraordinarily ...

  7. Nucleotide diversity and selection signature in the domesticated silkworm, Bombyx mori, and wild silkworm, Bombyx mandarina.

    Science.gov (United States)

    Guo, Yi; Shen, Yi-Hong; Sun, Wei; Kishino, Hirohisa; Xiang, Zhong-Huai; Zhang, Ze

    2011-01-01

    To investigate the patterns of nucleotide diversity in domesticated silkworm, Bombyx mori L. (Lepidoptera: Bombycidae) and its wild relative, Chinese wild silkworm, Bombyx mandarina Moore, we sequenced nine nuclear genes. Neutrality test and coalescent simulation for these genes were performed to look at bottleneck intensity and selection signature; linkage disequilibrium (LD) within and between loci was employed to investigate allele association. As a result, B. mori lost 33-49% of nucleotide diversity relative to wild silkworm, which is similar to the loss levels found in major cultivated crops. Diversity of B. mori is significantly lower than that of B. mandarina measured as π(total) (0.01166 vs. 0.1741) or θ(W)(0.01124 vs. 0.02206). Bottleneck intensity of domesticated silkworm is 1.5 (in terms of k = N(b) /d, N(b) -bottleneck population size; d-bottleneck duration) with different durations. Gene DefA showed signature of artificial selection by all analysis methods and might experience strong artificial selection in B. mori during domestication. For nine loci, both curves of LD decay rapidly within 200 bp and drop slowly when distance is > 200 bp, although that of B. mori decays slower than B. mandarina at loci investigated. However, LD could not be estimated at DefA in B. mori and at ER in both silkworms. Elevated LD observed in B. mori may be indicator of selection and demographic events.

  8. Horizontal gene transfer in silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Li Bin

    2011-05-01

    Full Text Available Abstract Background The domesticated silkworm, Bombyx mori, is the model insect for the order Lepidoptera, has economically important values, and has gained some representative behavioral characteristics compared to its wild ancestor. The genome of B. mori has been fully sequenced while function analysis of BmChi-h and BmSuc1 genes revealed that horizontal gene transfer (HGT maybe bestow a clear selective advantage to B. mori. However, the role of HGT in the evolutionary history of B. mori is largely unexplored. In this study, we compare the whole genome of B. mori with those of 382 prokaryotic and eukaryotic species to investigate the potential HGTs. Results Ten candidate HGT events were defined in B. mori by comprehensive sequence analysis using Maximum Likelihood and Bayesian method combining with EST checking. Phylogenetic analysis of the candidate HGT genes suggested that one HGT was plant-to- B. mori transfer while nine were bacteria-to- B. mori transfer. Furthermore, functional analysis based on expression, coexpression and related literature searching revealed that several HGT candidate genes have added important characters, such as resistance to pathogen, to B. mori. Conclusions Results from this study clearly demonstrated that HGTs play an important role in the evolution of B. mori although the number of HGT events in B. mori is in general smaller than those of microbes and other insects. In particular, interdomain HGTs in B. mori may give rise to functional, persistent, and possibly evolutionarily significant new genes.

  9. Subcelluar localization of orf126 of Bombyx mori nucleopolyhedrovirus

    African Journals Online (AJOL)

    In order to explore the mechanism of orf126 of Bombyx mori nucleopolyhedro virus, the subcellular localization of ORF126 was conducted. The egfp gene was fused with the C-terminal of orf126 genes, BmN cells were transfected with different plasmid DNA and the superinfection were performed at 12 h post transfection.

  10. Genome engineering and parthenocloning in the silkworm, Bombyx ...

    Indian Academy of Sciences (India)

    Genetic engineering of the silkworm, Bombyx mori, opens door to the production of new kinds of silk and to the use of silkworms as proteosynthetic bioreactors. The insertion of foreign genes into silkworm genome and the control of their expression by diverse promoters have become possible but are not yet efficient enough ...

  11. Genome engineering and parthenocloning in the silkworm, Bombyx ...

    Indian Academy of Sciences (India)

    2015-08-15

    Aug 15, 2015 ... Genetic engineering of the silkworm, Bombyx mori, opens door to the production of new kinds of silk and to the use of silkworms as ..... substrate suitable for the cocoon attachment, and eventually glues fibers to one another in ..... tion in silkworm silk glands requires cathepsin and chitinase of. Autographa ...

  12. RNA recognition motif (RRM)-containing proteins in Bombyx mori ...

    African Journals Online (AJOL)

    Bombyx mori is a model species of Lepidoptera whose genome sequence is available; however, its RRM-containing proteins have not been thoroughly analyzed. In this study, 134 RRM-containing protein genes in Drosophila melanogaster were used to perform BLAST search against B. mori genome- and EST databases.

  13. Efficient TALEN construction for Bombyx mori gene targeting

    Czech Academy of Sciences Publication Activity Database

    Takasu, Y.; Sajwan, Suresh; Daimon, T.; Osanai-Futahashi, M.; Uchino, K.; Sezutsu, H.; Tamura, T.; Žurovec, Michal

    2013-01-01

    Roč. 8, č. 9 (2013), e73458 E-ISSN 1932-6203 R&D Projects: GA ČR GAP305/10/2406 Grant - others:Japan Society for the Promotion of Science(JP) 23580083 Institutional support: RVO:60077344 Keywords : Bombyx mori Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.534, year: 2013 http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0073458

  14. Diapause prevention effect of Bombyx mori by dimethyl sulfoxide.

    Directory of Open Access Journals (Sweden)

    Takayuki Yamamoto

    Full Text Available HCl treatment has been, for about 80 years, the primary method for the prevention of entry into embryonic diapauses of Bombyx mori. This is because no method is as effective as the HCl treatment. In this study, we discovered that dimethyl sulfoxide (DMSO prevented entry into the diapause of the silkworm, Bombyx mori. The effect of diapause prevention was 78% as a result of treatment with 100% DMSO concentration, and the effect was comparable to that of the HCl treatment. In contrast, in the case of non-diapause eggs, hatchability was decreased by DMSO in a concentration-dependent manner. The effect of DMSO was restricted within 24 hours after oviposition of diapause eggs, and the critical period was slightly shorter than the effective period of the HCl treatment. DMSO analogs, such as dimethyl formamide (DMF and dimethyl sulfide (DMS, did little preventive effect against the diapause. Furthermore, we also investigated the permeation effects of chemical compounds by DMSO. When treated with an inhibitor of protein kinase CK2 (CK2 dissolved in DMSO, the prevention rate of the diapause was less than 40%. This means that the inhibition effect by the CK2 inhibitor was the inhibition of embryonic development after diapause prevention by DMSO. These data suggest that DMSO has the effects of preventing from entering into the diapause and permeation of chemicals into diapause eggs.

  15. Transcriptome sequencing and positive selected genes analysis of Bombyx mandarina.

    Directory of Open Access Journals (Sweden)

    Tingcai Cheng

    Full Text Available The wild silkworm Bombyx mandarina is widely believed to be an ancestor of the domesticated silkworm, Bombyx mori. Silkworms are often used as a model for studying the mechanism of species domestication. Here, we performed transcriptome sequencing of the wild silkworm using an Illumina HiSeq2000 platform. We produced 100,004,078 high-quality reads and assembled them into 50,773 contigs with an N50 length of 1764 bp and a mean length of 941.62 bp. A total of 33,759 unigenes were identified, with 12,805 annotated in the Nr database, 8273 in the Pfam database, and 9093 in the Swiss-Prot database. Expression profile analysis found significant differential expression of 1308 unigenes between the middle silk gland (MSG and posterior silk gland (PSG. Three sericin genes (sericin 1, sericin 2, and sericin 3 were expressed specifically in the MSG and three fibroin genes (fibroin-H, fibroin-L, and fibroin/P25 were expressed specifically in the PSG. In addition, 32,297 Single-nucleotide polymorphisms (SNPs and 361 insertion-deletions (INDELs were detected. Comparison with the domesticated silkworm p50/Dazao identified 5,295 orthologous genes, among which 400 might have experienced or to be experiencing positive selection by Ka/Ks analysis. These data and analyses presented here provide insights into silkworm domestication and an invaluable resource for wild silkworm genomics research.

  16. Transposable-element associated small RNAs in Bombyx mori genome.

    Directory of Open Access Journals (Sweden)

    Yimei Cai

    Full Text Available Small RNAs are a group of regulatory RNA molecules that control gene expression at transcriptional or post-transcriptional levels among eukaryotes. The silkworm, Bombyx mori L., genome harbors abundant repetitive sequences derived from families of retrotransposons and transposons, which together constitute almost half of the genome space and provide ample resource for biogenesis of the three major small RNA families. We systematically discovered transposable-element (TE-associated small RNAs in B. mori genome based on a deep RNA-sequencing strategy and the effort yielded 182, 788 and 4,990 TE-associated small RNAs in the miRNA, siRNA and piRNA species, respectively. Our analysis suggested that the three small RNA species preferentially associate with different TEs to create sequence and functional diversity, and we also show evidence that a Bombyx non-LTR retrotransposon, bm1645, alone contributes to the generation of TE-associated small RNAs in a very significant way. The fact that bm1645-associated small RNAs partially overlap with each other implies a possibility that this element may be modulated by different mechanisms to generate different products with diverse functions. Taken together, these discoveries expand the small RNA pool in B. mori genome and lead to new knowledge on the diversity and functional significance of TE-associated small RNAs.

  17. Cytotoxic compounds against cancer cells from Bombyx mori inoculated with Cordyceps militaris.

    Science.gov (United States)

    Qiu, Weitao; Wu, Jing; Choi, Jae-Hoon; Hirai, Hirofumi; Nishida, Hiroshi; Kawagishi, Hirokazu

    2017-06-01

    Two compounds, 3'-deoxyinosine and cordycepin, were isolated from Bombyx mori inoculated with Cordyceps militaris. In the bioassay examining cytotoxicity against cancer cells, both compounds showed toxicity against A549, PANC-1, and MCF-7 cancer cells.

  18. Discovery of anti-viral molecules and their vital functions in Bombyx mori.

    Science.gov (United States)

    Lü, Peng; Pan, Ye; Yang, Yanhua; Zhu, Feifei; Li, Chengjun; Guo, Zhongjian; Yao, Qin; Chen, Keping

    2018-02-14

    The silkworm Bombyx mori (B. mori), a lepidopteran model organism, has become an important model for molecular biology researches with its genome completely sequenced. Silkworms confront different types of virus diseases, mainly including those caused by Bombyx mori nucleopolyhedrovirus (BmNPV), Bombyx mori densovirus type 1 (BmDNV-1), Bombyx mori bidesovirus (BmBDV) which was termed as Bombyx mori densovirus type 2 (BmDNV-2) or Bombyx mori parvo-like virus (BmPLV) before in sericulture. B. mori offers excellent models to study the molecular mechanisms of insect innate immune responses to viruses. A variety of molecules and pathways have been identified to be involved in the immune responses in the silkworm to viruses, such as the antimicrobial peptides, prophenoloxidase-activating system, apoptosis, ROS, small RNA and related molecules. Here in this review, we summarize the current research advances in molecules involved in silkworm anti-virus pathways. Moreover, taking BmNPV as an example, we proposed a schematic model of molecules and pathways involved in silkworm immune responses against virus infection. We hope this review can facilitate further study of antiviral mechanisms in silkworm, and provide a reference for virus diseases in other organisms. Copyright © 2018 Elsevier Inc. All rights reserved.

  19. Recent progress in genome engineering techniques in the silkworm, Bombyx mori.

    Science.gov (United States)

    Daimon, Takaaki; Kiuchi, Takashi; Takasu, Yoko

    2014-01-01

    Rapid advances in genome engineering tools, such as zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the clustered regularly interspaced palindromic repeats/CRISPR-associated (CRISPR/Cas) system, have enabled efficient gene knockout experiments in a wide variety of organisms. Here, we review the recent progress in targeted gene disruption techniques in the silkworm, Bombyx mori. Although efficiency of targeted mutagenesis was very low in an early experiment using ZFNs, recent studies have shown that TALENs can induce highly efficient mutagenesis of desired target genes in Bombyx. Notably, mutation frequencies induced by TALENs can reach more than 50% of G0 gametes. Thus, TALENs can now be used as a standard tool for gene targeting studies, even when mutant phenotypes are unknown. We also propose guidelines for experimental design and strategy for knockout experiments in Bombyx. Genome editing technologies will greatly increase the usefulness of Bombyx as a model for lepidopteran insects, the major agricultural pests, and lead to sophisticated breeding of Bombyx for use in sericulture and biotechnology. © 2013 The Authors Development, Growth & Differentiation © 2013 Japanese Society of Developmental Biologists.

  20. Molecular Characterization of Endoplasmic Reticulum Oxidoreductin 1 from Bombyx mori

    Directory of Open Access Journals (Sweden)

    Minchul Seo

    2015-11-01

    Full Text Available We isolated a complementary DNA (cDNA clone encoding endoplasmic reticulum oxidoreductin 1 (bERO1, a specific oxidant of protein disulfide isomerase (PDI from Bombyx mori. This protein has a putative open reading frame (ORF of 489 amino acids and a predicted size of 57.4 kDa. Although bERO1 protein shares less than 57% amino acid sequence homology with other reported ERO1s, it contains two conserved redox active motifs, a Cys-X-X-X-X-Cys motif of N-terminal and Cys-X-X-Cys-X-X-Cys motif of C-terminal. Both motifs are typically present in ERO1 protein family members. The bEro1 mRNA expression was highest in posterior silk gland on the sixth day of the 5th instar larvae. Expression of bEro1 mRNA also markedly increased during endoplasmic reticulum (ER stress induced by stimulation with antimycin, calcium ionophore A23187, dithiothreitol, H2O2, monencin, and tunicamycin. In addition, expression levels of bEro1 exactly coincided with that of bPdi. This is the first result suggesting that bERO1 plays an essential role in ER quality control through the combined activities of bERO1 and bPDI as a catalyst of protein folding in the ER and sustaining cellular redox homeostasis.

  1. Metabonomic analysis of Bombyx mori (Heterocera: Bombysidae) treated with acetaminophen.

    Science.gov (United States)

    Yin, W M; Xu, X; He, Y; Wei, G B; Sima, Y H; Shi-Qing, Xu

    2014-01-01

    The feasibility of using Bombyx mori as model animal is attracting more attention. Whether the effect of drugs on the metabolite profiling was consistent with those in mammals was an aspect to evaluate the feasibility of B. mori as model animal. In this study, we used acetaminophen to treat Dazao fifth-instar B. mori, and its metabolites in hemolymph were detected by gas chromatography-mass spectrometry. The corresponding data were processed and analyzed by total model analysis, principal component analysis, partial least squares-discriminant analysis, orthogonal partial least squares-discriminant analysis, and finally, the difference metabolites between acetaminophen group and control group were selected and identified by our reference material database and the National Institute of Standard and Technology database. The results showed that acetaminophen administration induced elevation of metabolites related to energy source, the intermediate of cholesterol synthesis, and the metabolites related to melanization and also induced the decrease of metabolites in pathway of Krebs cycle, the cholesterol, and sitosterol, which suggested that acetaminophen administration inhibited energy metabolism and promoted the expenditure and imbalance of hormone and melanization. © The Author 2014. Published by Oxford University Press on behalf of the Entomological Society of America.

  2. A characterization of structural proteins expressed by Bombyx mori bidensovirus.

    Science.gov (United States)

    Lü, Peng; Xing, Yali; Hu, Zhaoyang; Yang, Yanhua; Pan, Ye; Chen, Kangmin; Zhu, Feifei; Zhou, Yajing; Chen, Keping; Yao, Qin

    2017-03-01

    Bombyx mori bidensiovirus (BmBDV) is a species of Bidensovirus that has been was placed into a new genus within the new family Bidnaviridae by the International Committee on Taxonomy of Viruses. BmBDV causes fatal flacherie disease in silkworms, which causes large losses to the sericulture industry. BmBDV contains two sets of complementary linear single-stranded DNAs of approximately 6.5kb (viral DNA 1, VD1) and 6.0kb (viral DNA 2, VD2). VD1 and VD2 are encapsidated in separate icosahedral non-enveloped capsids, which are similar in size and shape. However, the strategies used to express BmBDV structural proteins remains unclear. In this work, a total of six structural proteins were separated by two-dimensional electrophoresis and shown to be encoded by the BmBDV VP gene via mass spectrometry. The transmission electron microscopy results showed that co-expression of the BmBDV VP and SP structural proteins in Spodoptera frugiperda sf9 cells resulted in the formation of 22-24nm virus-like particles. Furthermore, a mutation of the major structural protein-encoding VP gene, in which the second in-frame ATG codon was mutated to GCG, abrogated the production of several structural proteins, indicating that this strategy of expressing BmBDV VP is dependent on a leaky scanning translation mechanism. Copyright © 2016. Published by Elsevier Inc.

  3. Study on the radiation sterile mechanism of China mulberry wild silkworm (Bombyx mandorina Moore)

    International Nuclear Information System (INIS)

    Yang Rongxin; Xia Darong; Gu Weiping; Meng Zhiqi; Weng Hongbiao

    2000-01-01

    The sterile mechanism of China mulberry wild silkworm (Bombyx mandorina Moore) was studied on the chromosome structure variation by means of 60 Co γ-rays irradiation of the pupae. The dose of sub-sterilization for the wild silkworm was 250 Gy, under which the females of present generation were whole sterilized and the males were sub-sterilized. The chromosome structure variation such as deficiency and inversion of larva reproductive cells could be obviously observed. Most of cells with the abnormal chromosomes were sterilized, which was the main reason or radiation sterilization of Bombyx mandorina Moore

  4. Fate mapping of the silkworm, Bombyx mori, using localized UV irradiation of the egg at fertilization

    International Nuclear Information System (INIS)

    Myohara, Maroko

    1994-01-01

    Bombyx eggs at the fertilization stage (0-2 hours after oviposition) were irradiated with a scanning UV-laser microbeam (355 nm) over an area of about 1% of the total egg surface. In spite of absence of nuclei or cells at the irradiated sites, larvae from treated eggs showed localized cuticle defects in the integument. The location and frequency of the defects within the cuticular pattern correlated closely to the site of irradiation both in the anteroposterior and the dorsoventral direction. Based on the correlation, presumptive regions for each larval segment were located and a fate map of the Bombyx egg was established. (author)

  5. The dynamic landscape of gene regulation during Bombyx mori oogenesis.

    Science.gov (United States)

    Zhang, Qiang; Sun, Wei; Sun, Bang-Yong; Xiao, Yang; Zhang, Ze

    2017-09-11

    Oogenesis in the domestic silkworm (Bombyx mori) is a complex process involving previtellogenesis, vitellogenesis and choriogenesis. During this process, follicles show drastic morphological and physiological changes. However, the genome-wide regulatory profiles of gene expression during oogenesis remain to be determined. In this study, we obtained time-series transcriptome data and used these data to reveal the dynamic landscape of gene regulation during oogenesis. A total of 1932 genes were identified to be differentially expressed among different stages, most of which occurred during the transition from late vitellogenesis to early choriogenesis. Using weighted gene co-expression network analysis, we identified six stage-specific gene modules that correspond to multiple regulatory pathways. Strikingly, the biosynthesis pathway of the molting hormone 20-hydroxyecdysone (20E) was enriched in one of the modules. Further analysis showed that the ecdysteroid 20-hydroxylase gene (CYP314A1) of steroidgenesis genes was mainly expressed in previtellogenesis and early vitellogenesis. However, the 20E-inactivated genes, particularly the ecdysteroid 26-hydroxylase encoding gene (Cyp18a1), were highly expressed in late vitellogenesis. These distinct expression patterns between 20E synthesis and catabolism-related genes might ensure the rapid decline of the hormone titer at the transition point from vitellogenesis to choriogenesis. In addition, we compared landscapes of gene regulation between silkworm (Lepidoptera) and fruit fly (Diptera) oogeneses. Our results show that there is some consensus in the modules of gene co-expression during oogenesis in these insects. The data presented in this study provide new insights into the regulatory mechanisms underlying oogenesis in insects with polytrophic meroistic ovaries. The results also provide clues for further investigating the roles of epigenetic reconfiguration and circadian rhythm in insect oogenesis.

  6. Anterior and posterior centers jointly regulate Bombyx embryo body segmentation.

    Science.gov (United States)

    Nakao, Hajime

    2012-11-15

    Insect embryo segmentation is largely divided into long and short germ types. In the long germ type, each segment primordium is represented on a large embryonic rudiment of the blastoderm, and segmental patterning occurs nearly simultaneously in the syncytium. In the short germ type, however, only anterior segments are represented in the small embryonic rudiment, usually located on the egg posterior, and the rest of the segments are added sequentially from the posterior growth zone in a cellular context. The long germ type is thought to have evolved from the short germ type. It is proposed that this transition, which appears to have occurred multiple times over the course of evolution, was realized through the acquisition of a localized anterior instruction center. Here, I examined the early segmentation process in the silkmoth Bombyx mori, a lepidopteran insect, in which the mechanisms of anterior-posterior (AP) axis formation have not been well analyzed. In this insect, both the long germ and short germ features have been reported. The mRNAs for two key genes involved in insect AP axis formation, orthodenticle (Bm-otd) and caudal (Bm-cad), are localized maternally in the germ anlage, where they act as anterior and posterior instruction centers, respectively. RNAi studies indicate that, while Bm-cad affects the formation of all the even skipped (Bm-eve) stripes, there is also anterior Bm-eve stripe formation activity that involves Bm-otd. Thus, there is redundancy in Bm-eve stripe formation activity that must be coordinated. Some genetic interactions, identified either experimentally or hypothetically, are also introduced, which might enable robust AP formation in this organism. Copyright © 2012 Elsevier Inc. All rights reserved.

  7. Development of a standard acute dietary toxicity test for the silkworm (Bombyx mori L.)

    NARCIS (Netherlands)

    Sun, X.; Valk, H.; Jiang, H.; Wang, X.; Yuan, S.; Zhang, Y.; Roessink, I.; Gao, X.

    2012-01-01

    Larvae of the silkworm (Bombyx mod L.) may be exposed to pesticide residues on the leaves of their food plant, the mulberry tree (Morus spp.), which can lead to adverse effects on silk production. A new acute dietary toxicity test method was evaluated as the basis for pesticide risk assessment. A

  8. Cloning and homologic analysis of Tpn I gene in silkworm Bombyx ...

    African Journals Online (AJOL)

    Cloning and homologic analysis of Tpn I gene in silkworm Bombyx mori. Y Zhao, Yao Q, X Tang, Q Wang, H Yin, Z Hu, J Lu, K Chen. Abstract. The troponin complex is composed of three subunits, Troponin C (the calcium sensor component) and Troponin T and I (structural proteins). Tpn C is encoded by multiple genes in ...

  9. The p10 gene of Bombyx mori nucleopolyhedrosis virus encodes a ...

    Indian Academy of Sciences (India)

    In baculovirus-based high-level expression of cloned foreign genes, the viral very late gene promoters of polyhedrin (polh) and p10 are extensively exploited. Here we report the cloning and characterization of the p10 gene from a local isolate of Bombyx mori nucleopolyhedrosis virus (BmNPV). The gene harbours a 213-bp ...

  10. Timeless is a critical gene in the diapause of silkworm, Bombyx mori ...

    African Journals Online (AJOL)

    In this research, the authors investigated the linkage of diapause and the regulation of circadian rhythm by inspecting the expression profiles of a circadian clock gene timeless (tim) under diapause-inducing temperature and photoperiod in Bombyx mori. The results show that the level of tim mRNA was higher in 25LL than in ...

  11. Differential expression of early viral gene BmORF51 in Bombyx mori ...

    African Journals Online (AJOL)

    USER

    2010-06-21

    Jun 21, 2010 ... Differential expression of early viral gene BmORF51 in. Bombyx mori nucleopolyhedrovirus infection of resistant and susceptible silkworms. Feng Lin, Qin Yao, Huiqing Chen, Yang Zhou and KePing Chen*. Institute of Life Sciences, Jiangsu University, 301 Xuefu Road, Zhenjiang 212013, P. R. China.

  12. Cloning and expression of a portion of Bombyx Mori Ser/2 gene

    OpenAIRE

    STAŠKOVÁ, Tereza

    2009-01-01

    A 897 bp section of Bombyx mori Ser-2 gene was amplified, cloned into a bacterial expression vector, and used to prepare a recombinant sericin-like protein. The protein is expected to contain domain supporting growth of mammalian cells. It will be used in assays verifying this assumption; if confirmed, recombinant protein will be considered for preparations of scaffold guiding tissue reconstruction.

  13. The wings of Bombyx mori develop from larval discs exhibiting an ...

    Indian Academy of Sciences (India)

    Lepidopteran insects present a complex organization of appendages which develop by various mechanisms. In the mulberry silkworm, Bombyx mori a pair of meso- and meta-thoracic discs located on either side in the larvae gives rise to the corresponding fore- and hind-wings of the adult. These discs do not experience ...

  14. Silkworm (Bombyx mori) hemocytes do not produce reactive oxygen metabolites as a part of defense mechanisms

    Czech Academy of Sciences Publication Activity Database

    Hyršl, P.; Číž, Milan; Kubala, Lukáš; Lojek, Antonín

    2004-01-01

    Roč. 49, č. 3 (2004), s. 315-319 ISSN 0015-5632 R&D Projects: GA AV ČR IBS5004009 Institutional research plan: CEZ:AV0Z5004920 Keywords : hemocytes * Bombyx mori * reactive oxygen species Subject RIV: BO - Biophysics Impact factor: 1.034, year: 2004

  15. Annotation and expression of carboxylesterases in the silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Li Wen-Le

    2009-11-01

    Full Text Available Abstract Background Carboxylesterase is a multifunctional superfamily and ubiquitous in all living organisms, including animals, plants, insects, and microbes. It plays important roles in xenobiotic detoxification, and pheromone degradation, neurogenesis and regulating development. Previous studies mainly used Dipteran Drosophila and mosquitoes as model organisms to investigate the roles of the insect COEs in insecticide resistance. However, genome-wide characterization of COEs in phytophagous insects and comparative analysis remain to be performed. Results Based on the newly assembled genome sequence, 76 putative COEs were identified in Bombyx mori. Relative to other Dipteran and Hymenopteran insects, alpha-esterases were significantly expanded in the silkworm. Genomics analysis suggested that BmCOEs showed chromosome preferable distribution and 55% of which were tandem arranged. Sixty-one BmCOEs were transcribed based on cDNA/ESTs and microarray data. Generally, most of the COEs showed tissue specific expressions and expression level between male and female did not display obvious differences. Three main patterns could be classified, i.e. midgut-, head and integument-, and silk gland-specific expressions. Midgut is the first barrier of xenobiotics peroral toxicity, in which COEs may be involved in eliminating secondary metabolites of mulberry leaves and contaminants of insecticides in diet. For head and integument-class, most of the members were homologous to odorant-degrading enzyme (ODE and antennal esterase. RT-PCR verified that the ODE-like esterases were also highly expressed in larvae antenna and maxilla, and thus they may play important roles in degradation of plant volatiles or other xenobiotics. Conclusion B. mori has the largest number of insect COE genes characterized to date. Comparative genomic analysis suggested that the gene expansion mainly occurred in silkworm alpha-esterases. Expression evidence indicated that the expanded

  16. Respiratory allergy to moth: the importance of sensitization to Bombyx mori in children with asthma and rhinitis.

    Science.gov (United States)

    Araujo, Laura M L; Rosário Filho, Nelson A; Riedi, Carlos A

    2014-01-01

    this study aimed to prepare a silkworm moth (Bombyx mori) antigenic extract and to perform skin prick tests with this extract in patients with allergic respiratory diseases; to evaluate serum specific immunoglobulin E (IgE) to Bombyx mori using ImmunoCAP® system and to report the frequency of positivity between the two methods and with clinical data. this was a cross-sectional study with 99 children and adolescents diagnosed with asthma and/or allergic rhinitis, who had skin reactivity to at least one of the six aeroallergens tested. Clinical data were evaluated: skin prick tests with Bombyx mori in-house extract, and total and specific IgE analysis using ImmunoCAP® were performed. the frequency of Bombyx mori specific IgE was found to be 52.5% and 60% using the skin prick test and ImmunoCAP®, respectively. An association between a positive skin test for Bombyx mori and the presence of allergic rhinitis, atopic dermatitis, and urticaria was observed, but the same was not true for asthma or allergic conjunctivitis. There was no relation with the severity of asthma or rhinitis symptoms. a high frequency of sensitization to Bombyx mori was observed in a selected population of patients with respiratory allergic diseases in the city of Curitiba, state of Paraná, Brazil. The extract prepared from the wings of this moth species is effective in demonstrating this sensitivity. Copyright © 2013 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. All rights reserved.

  17. Respiratory allergy to moth: the importance of sensitization to Bombyx mori in children with asthma and rhinitis ,

    Directory of Open Access Journals (Sweden)

    Laura M.L. Araujo

    2014-04-01

    Full Text Available OBJECTIVE:this study aimed to prepare a silkworm moth (Bombyx mori antigenic extract and to perform skin prick tests with this extract in patients with allergic respiratory diseases; to evaluate serum specific immunoglobulin E (IgE to Bombyx mori using ImmunoCAP(r system and to report the frequency of positivity between the two methods and with clinical data.METHODS:this was a cross-sectional study with 99 children and adolescents diagnosed with asthma and/or allergic rhinitis, who had skin reactivity to at least one of the six aeroallergens tested. Clinical data were evaluated: skin prick tests with Bombyx mori in-house extract, and total and specific IgE analysis using ImmunoCAP(r were performed.RESULTS:the frequency of Bombyx mori specific IgE was found to be 52.5% and 60% using the skin prick test and ImmunoCAP(r, respectively. An association between a positive skin test for Bombyx mori and the presence of allergic rhinitis, atopic dermatitis, and urticaria was observed, but the same was not true for asthma or allergic conjunctivitis. There was no relation with the severity of asthma or rhinitis symptoms.CONCLUSIONS:a high frequency of sensitization to Bombyx mori was observed in a selected population of patients with respiratory allergic diseases in the city of Curitiba, state of Paraná, Brazil. The extract prepared from the wings of this moth species is effective in demonstrating this sensitivity.

  18. Evolutionarily conserved repulsive guidance role of slit in the silkworm Bombyx mori.

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    Qi Yu

    Full Text Available Axon guidance molecule Slit is critical for the axon repulsion in neural tissues, which is evolutionarily conserved from planarians to humans. However, the function of Slit in the silkworm Bombyx mori was unknown. Here we showed that the structure of Bombyx mori Slit (BmSlit was different from that in most other species in its C-terminal sequence. BmSlit was localized in the midline glial cell, the neuropil, the tendon cell, the muscle and the silk gland and colocalized with BmRobo1 in the neuropil, the muscle and the silk gland. Knock-down of Bmslit by RNA interference (RNAi resulted in abnormal development of axons and muscles. Our results suggest that BmSlit has a repulsive role in axon guidance and muscle migration. Moreover, the localization of BmSlit in the silk gland argues for its important function in the development of the silk gland.

  19. Silvernanotherapy on the viral borne disease of silkworm Bombyx mori L.

    Science.gov (United States)

    Govindaraju, K.; Tamilselvan, S.; Kiruthiga, V.; Singaravelu, G.

    2011-12-01

    Antiviral assays of chemically and biologically synthesized silver nanoparticles were made against BmNPV ( Bombyx mori Nuclear Polyhedrosis Virus). Reduction of silver ions by sodium citrate and Spirulina platensis led to the formation of spherical silver nanoparticles of 40-60 and 7-16 nm size. Single cell protein ( Spirulina platensis)-synthesized silver nanoparticles showed the strongest antiviral activity. Immunological studies made on the silkworm Bombyx mori disclosed that a significant increase in the total hemocyte count and differential hemocyte count due to S. platensis-synthesized silver nanoparticles supplementation. Improvement in the defense mechanism was noticed from the strengthened peritrophic membrane of the digestive tract and the increased total protein. Overall, the results presented illustrate that single cell protein-synthesized silver nanoparticles supplementation is effective in controlling viral-borne diseases of the silkworm.

  20. The Effect of Some Additives on the Bioproductive Performances of SilkWorm Bombyx mori L.

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    Mihai Benţea

    2011-05-01

    Full Text Available Nowdays, worldwide in sericulture domain, researches are aimed to find new efficient alternatives for the feeding of mulberry silk worm through the use of some fodder additives which allow the improvement of bioproductive parameters and silk line quality. The present work represents a review of the use of such substances (mineral and vitamin additives and of the experiments carried out to point out their effects on silkworms (Bombyx mori L.. Research conducted worldwide revealed the fact that the use of mineral and vitamin additives in silkworm Bombyx mori L., improve the growth rate of larvae, protein and mineral content of haemolymph, the protein content, DNA and RNA content of the sericigen glands, and the quality parameters of cocoon and silk line. Researches showed that the use of these additives doesn’t affect the quality of the silk.

  1. Activity patterns of neurosecretory cells releasing pheromonotropic neuropeptides in the moth Bombyx mori

    OpenAIRE

    Ichikawa, Toshio

    1998-01-01

    Short- and long-term firing patterns of neurosecretory cells releasing pheromonotropic neuropeptides in the silkworm moth Bombyx mori were examined. The cells showed three types of rhythmic changes in firing activity. Bursting activities with an interval of several seconds were synchronized with rhythmic abdominal motions for calling behavior. A slow fluctuation in firing activity over a period of several minutes depended on cyclic alternations of the flow of hemolymph. The electrical activit...

  2. Physicochemical Properties of Meat Batter Added with Edible Silkworm Pupae (Bombyx mori) and Transglutaminase

    OpenAIRE

    Park, Yoo-Sun; Choi, Yun-Sang; Hwang, Ko-Eun; Kim, Tae-Kyung; Lee, Cheol-Won; Shin, Dong-Min; Han, Sung Gu

    2017-01-01

    This study was conducted to investigate the physicochemical properties of meat batters prepared with fresh pork meat, back fat, water, and salt and formulated with three different amounts (5%, 10%, and 15%) of silkworm pupae (Bombyx mori) powder and transglutaminase (TG). Meat batters formulated with silkworm pupae powder showed significantly higher contents of protein and ash than control batter. Addition of silkworm pupae to batter also showed significantly lower cooking loss than the contr...

  3. Nutrigenetic Screening Strains of the Mulberry Silkworm, Bombyx mori, for Nutritional Efficiency

    OpenAIRE

    Ramesha, Chinnaswamy; Lakshmi, Hothur; Kumari, Savarapu Sugnana; Anuradha, Chevva M.; Kumar, Chitta Suresh

    2012-01-01

    The activity of sericulture is declining due the reduction of mulberry production area in sericulture practicing countries lead to adverse effects on silkworm rearing and cocoon production. Screening for nutrigenetic traits in silkworm, Bombyx mori L. (Lepidoptera: Bombycidae) is an essential prerequisite for better understanding and development of nutritionally efficient breeds/hybrids, which show less food consumption with higher efficiency conversion. The aim of this study was to identify ...

  4. Application of artificial insemination technique to eupyrene and/or apyrene sperm in Bombyx mori

    OpenAIRE

    Sahara, Ken; Takemura, Yoko

    2003-01-01

    The silkworm, Bombyx mori, has a dimorphic sperm system. The eupyrene sperm is the sperm to fertilize eggs and the apyrene sperm plays a crucial role for assisting fertilization. Heat-treated (331C for 96h) Daizo (DH) males, one of the strains in the silkworm, produce only eupyrene sperm, while in triploid males only apyrene sperm are functional. Though both types of males are found to be sterile, double copulation of the two males with a single female greatly increases fertili...

  5. Two Dimensional Electrophoresis of Galactosidase Relating to the Disappearance of Bombyx Lectin Activity

    OpenAIRE

    カトウ, ヤスオ; Yasuo, Kato

    2004-01-01

    "Two dimensional polyacrylamide gel electroporesis (2 D-PAGE) analysis on the haemolymph of Bombyx mori was performed using the Mini-PROTEAN mini tube gel two dimensional polyacrylamide gel electrophoresis system (Bio-Rad Laboratories, Inc.). The result on various electrophoretical conditions using the haemolymph-protein showed the possibility that the haemolymph-protein was separated actually by means of this method. Moreover, the result of 2 D-PAGE analysis on Fraction II obtained by gel fi...

  6. Enhancer activity of Helitron in sericin-1 gene promoter from Bombyx mori.

    Science.gov (United States)

    Huang, Ke; Li, Chun-Feng; Wu, Jie; Wei, Jun-Hong; Zou, Yong; Han, Min-Jin; Zhou, Ze-Yang

    2016-06-01

    Sericin is a kind of water-soluble protein expressed specifically in the middle silk gland of Bombyx mori. When the sericin-1 gene promoter was cloned and a transgenic vector was constructed to express a foreign protein, a specific Helitron, Bmhel-8, was identified in the sericin-1 gene promoter sequence in some genotypes of Bombyx mori and Bombyx mandarina. Given that the Bmhel-8 Helitron transposon was present only in some genotypes, it could be the source of allelic variation in the sericin-1 promoter. The length of the sericin-1 promoter sequence is approximately 1063 or 643 bp. The larger size of the sequence or allele is ascribed to the presence of Bmhel-8. Silkworm genotypes can be homozygous for either the shorter or larger promoter sequence or heterozygous, containing both alleles. Bmhel-8 in the sericin-1 promoter exhibits enhancer activity, as demonstrated by a dual-luciferase reporter system in BmE cell lines. Furthermore, Bmhel-8 displays enhancer activity in a sericin-1 promoter-driven gene expression system but does not regulate the tissue-specific expression of sericin-1. © 2016 Institute of Zoology, Chinese Academy of Sciences.

  7. [Cloning and structural analysis of MLP in the silkworm, Bombyx mori].

    Science.gov (United States)

    Liu, Yan; Niu, Bao-Long; Weng, Hong-Biao; Shen, Wei-Feng; He, Li-Hua; Qi, Xiao-Peng; Meng, Zhi-Qi

    2007-09-01

    The LIM domain is found in a wide variety of eukaryotic proteins that regulate gene expression and cell differentiation during development. Muscle LIM protein (MLP) gene in Bombyx mori has been cloned by blasting its EST database and PCR test in present report. The resulting sequence covers 2 327 bp of cDNA (GenBank accession No. DQ311195). It has a complete open reading fragment and encodes a 494 amino acid protein. Genomic DNA sequence contains 11 exons and 10 introns, with intron splicing following the GT-AG rule. M.W. and PI of the predicted MLP in Bombyx mori are 53.03 kDa and 8.29 respectively. A single LIM domain linked to a glyscine-rich region is found in a previously deposited LIM protein (AAR23823) in Bombyx mori. MLP identified in this report encodes a protein with five tandem LIM-glycine modules. The two LIM proteins could be produced by alternative splicing and both are probably involved in muscle cell differentiation. This work provides foundation for further research on the in vivo function of MLP.

  8. Molecular cloning of a functional allatostatin gut/brain receptor and an allatostatin preprohormone from the silkworm Bombyx mori

    DEFF Research Database (Denmark)

    Secher, Thomas; Lenz, C; Cazzamali, G

    2001-01-01

    on the addition of 4 x 10(-9)M Bombyx A-type allatostatins with a second messenger cascade (measured as bioluminescence), showing that BAR is a functional A-type allatostatin receptor. Southern blots suggest that Bombyx has at least one other BAR-related gene in addition to the BAR gene described in this paper....... Northern blots and quantitative reverse transcriptase-polymerase chain reaction of different larval tissues show that BAR mRNA is mainly expressed in the gut and to a much lesser extent in the brain. To our knowledge, this is the first report on the molecular cloning and functional expression of an insect...

  9. Comparative Studies of Two-Dimensional Electrophoresis on Galactosidase Relating to Bombyx Lectin Activity

    OpenAIRE

    加藤, 靖夫; カトウ, ヤスオ; Yasuo, Kato

    2005-01-01

    "Comparative two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) analysis on the haemolymph of the domesticated silkworm, Bombyx mori and Fraction II obtained by gel filtration from the haemolymph of B. mori was performed using the 2-D mini-slab system (Atto Co.) (the first method of 2-D PAGE) and the Mini-PROTEAN mini tube gel 2-D PAGE system (Bio-Rad Laboratories, Inc.) (the second method). Moreover, two-dimensionnal electrophoresis analysis on standard β-galactosidase, grade III ...

  10. Combined Effect of Cameo2 and CBP on the Cellular Uptake of Lutein in the Silkworm, Bombyx mori

    Science.gov (United States)

    Dong, Xiao-Long; Chai, Chun-Li; Pan, Cai-Xia; Tang, Hui; Chen, Yan-Hong; Dai, Fang-Yin; Pan, Min-Hui; Lu, Cheng

    2014-01-01

    Formation of yellow-red color cocoons in the silkworm, Bombyx mori, occurs as the result of the selective delivery of carotenoids from the midgut to the silk gland via the hemolymph. This process of pigment transport is thought to be mediated by specific cellular carotenoids carrier proteins. Previous studies indicated that two proteins, Cameo2 and CBP, are associated with the selective transport of lutein from the midgut into the silk gland in Bombyx mori. However, the exact roles of Cameo2 and CBP during the uptake and transport of carotenoids are still unknown. In this study, we investigated the respective contributions of these two proteins to lutein and β-carotene transport in Bombyx mori as well as commercial cell-line. We found that tissues, expressed both Cameo2 and CBP, accumulate lutein. Cells, co-expressed Cameo2 and CBP, absorb 2 fold more lutein (Plutein was concentration-dependent and reached saturation. From immunofluorescence staining, confocal microscopy observation and western blot analysis, Cameo2 was localized at the membrane and CBP was expressed in the cytosol. What’s more, bimolecular fluorescence complementation analysis showed that these two proteins directly interacted at cellular level. Therefore, Cameo2 and CBP are necessarily expressed in midguts and silk glands for lutein uptake in Bombyx mori. Cameo2 and CBP, as the membrane protein and the cytosol protein, respectively, have the combined effect to facilitate the cellular uptake of lutein. PMID:24475153

  11. Respiratory allergy to moth: the importance of sensitization to Bombyx mori in children with asthma and rhinitis

    Directory of Open Access Journals (Sweden)

    Laura M.L. Araujo

    2014-03-01

    Conclusions: a high frequency of sensitization to Bombyx mori was observed in a selected population of patients with respiratory allergic diseases in the city of Curitiba, state of Paraná, Brazil. The extract prepared from the wings of this moth species is effective in demonstrating this sensitivity.

  12. Identification of Tyrosine Kinase Src Responsible for Antimicrobial Peptides Production in Bombyx mori.

    Science.gov (United States)

    Li, Xianyang; Hua, Xiaoting; Song, Liang; Xia, Qingyou; Wang, Fei

    2017-01-01

    Src is a non-receptor protein tyrosine kinase ubiquitously expressed in animals. It is involved in various cellular processes, including the innate immune response in mammals. However, less is known about the function of insect Src. Here we presented a homologue of Src in silkworm (Bombyx mori), named as BmSrc by phylogenetic analysis, homologous comparison and domain prediction. BmSrc contains the conserved phosphorylation residues and possesses tyrosine kinase activity. The expression pattern of BmSrc mRNA was specific in developmental stages and tissues. The highest expression of BmSrc was detected in moth stage, and the gonads showed the highest expression during larval stage. We then found over-expression of BmSrc in BmE cell resulted in an increase of p38 mitogen-activated protein kinase (p38 MAPK) and Akt phosphorylation but a decrease in extracellular signal-regulated kinase (ERK) phosphorylation. Finally, we demonstrated that BmSrc promoted the production of antimicrobial peptides (AMPs). These results implied that BmSrc is involved in immune response of silkworm possibly through activating p38 MAPK and Akt signaling pathway. Our study may provide reference for further investigation of the biological function of BmSrc in Bombyx mori. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  13. Identification and quantification and antioxidant activity of flavonoids in different strains of silk cocoon, Bombyx mori.

    Science.gov (United States)

    Napavichayanun, Supamas; Lutz, Oliver; Fischnaller, Martin; Jakschitz, Thomas; Bonn, Günther; Aramwit, Pornanong

    2017-10-01

    Silk cocoon is produced from silkworm (Bombyx mori) to protect itself from outer environment. Various strains of cocoon contain different forms and amounts of flavonoids, which may affect on their antioxidant activity. Moreover, the extraction method would influence the amount of flavonoids extracted. Therefore, the objectives of this study were to identify and quantify the flavonoids in 3 strains of bivoltine Bombyx mori silk cocoon (Chul 1/1; white cocoon, Chul 3/2; greenish cocoon, and Chul 4/2; yellow cocoon) extracted by 6 different solvents including acetone, ethyl acetate, dimethyl sulfoxide (DMSO), ethanol, methanol, and purified water. The flavonoids extracted were identified and quantified by liquid chromatography-mass spectrometry (LC-MS). The antioxidant activity of flavonoids extracted was also investigated by visible spectroscopy at 517 nm. The results showed that Chul 3/2 silk cocoon contained the highest amount of flavonoids. Purified water seemed to be the best solvent that preserved most antioxidant activity of the flavonoids extracted. Flavonoids in Chul 1/1 and Chul 4/2 silk cocoon were rarely found, however they contained some antioxidant activities. The data from this study can provide basic information for flavonoid extraction from silk cocoon which can also apply for other flavonoid-containing natural biomaterials. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Identification and analysis of YELLOW protein family genes in the silkworm, Bombyx mori

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    Yi Yong-Zhu

    2006-08-01

    Full Text Available Abstract Background The major royal jelly proteins/yellow (MRJP/YELLOW family possesses several physiological and chemical functions in the development of Apis mellifera and Drosophila melanogaster. Each protein of the family has a conserved domain named MRJP. However, there is no report of MRJP/YELLOW family proteins in the Lepidoptera. Results Using the YELLOW protein sequence in Drosophila melanogaster to BLAST silkworm EST database, we found a gene family composed of seven members with a conserved MRJP domain each and named it YELLOW protein family of Bombyx mori. We completed the cDNA sequences with RACE method. The protein of each member possesses a MRJP domain and a putative cleavable signal peptide consisting of a hydrophobic sequence. In view of genetic evolution, the whole Bm YELLOW protein family composes a monophyletic group, which is distinctly separate from Drosophila melanogaster and Apis mellifera. We then showed the tissue expression profiles of Bm YELLOW protein family genes by RT-PCR. Conclusion A Bombyx mori YELLOW protein family is found to be composed of at least seven members. The low homogeneity and unique pattern of gene expression by each member among the family ensure us to prophesy that the members of Bm YELLOW protein family would play some important physiological functions in silkworm development.

  15. Identification and Analysis of the SET-Domain Family in Silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Hailong Zhao

    2015-01-01

    Full Text Available As an important economic insect, Bombyx mori is also a useful model organism for lepidopteran insect. SET-domain-containing proteins belong to a group of enzymes named after a common domain that utilizes the cofactor S-adenosyl-L-methionine (SAM to achieve methylation of its substrates. Many SET-domain-containing proteins have been shown to display catalytic activity towards particular lysine residues on histones, but emerging evidence also indicates that various nonhistone proteins are specifically targeted by this clade of enzymes. To explore their diverse functions of SET-domain superfamily in insect, we identified, cloned, and analyzed the SET-domains proteins in silkworm, Bombyx mori. Firstly, 24 genes containing SET domain from silkworm genome were characterized and 17 of them belonged to six subfamilies of SUV39, SET1, SET2, SUV4-20, EZ, and SMYD. Secondly, SET domains of silkworm SET-domain family were intraspecifically and interspecifically conserved, especially for the catalytic core “NHSC” motif, substrate binding site, and catalytic site in the SET domain. Lastly, further analyses indicated that silkworm SET-domain gene BmSu(var3-9 owned different characterization and expression profiles compared to other invertebrates. Overall, our results provide a new insight into the functional and evolutionary features of SET-domain family.

  16. Cells of the rectum of Bombyx mori affected by experimental inoculation with Alphabaculovirus.

    Science.gov (United States)

    Vessaro-Silva, S A; Brancalhão, R M C; Baggio, M P D; Ribeiro, L F C

    2014-08-29

    Bombyx mori is an insect whose cocoon is used in the sericulture industry, which is an important activity in parts of southern Brazil. When parasitized by Bombyx mori nucleopolyhedrovirus (BmNPV) of the genus Alphabaculovirus (alphaBV), it develops nuclear polyhedrosis disease. In Brazil, an alphaBV was isolated from larvae of B. mori and various target tissues were identified. However, how this geographic viral isolate affects the rectum of silk moths was unknown. The rectum, a component of the cryptonephric system, acts to absorb water and mineral salts, and its importance for the metabolic balance of insects provoked interest in analyzing how it is affected by BmMNPV infection. Fifth instar B. mori larvae were inoculated with a viral suspension, and from the second to the ninth day post-inoculation, segments of the rectum (anterior and anal canal) were examined using light microscopy. The cryptonephric epithelial cells in the anterior region revealed no evidence of infection. However, from the fifth day post-inoculation, cells of the anal canal showed cytopathologies characteristic of alphaBV. Infection of the anal canal and other surrounding tissues led to tissue disorganization, with accumulation of polyhedra in the perinephric space and compartmentalization of the cryptonephric system, promoting changes in the fecal pellets, signalling physiological changes. These observations contribute to our understanding of the infectious cycle of BmMNPV in B. mori.

  17. Characterization of an entomopathogenic fungi target integument protein, Bombyx mori single domain von Willebrand factor type C, in the silkworm, Bombyx mori.

    Science.gov (United States)

    Han, F; Lu, A; Yuan, Y; Huang, W; Beerntsen, B T; Huang, J; Ling, E

    2017-06-01

    The insect cuticle works as the first line of defence to protect insects from pathogenic infections and water evaporation. However, the old cuticle must be shed in order to enter the next developmental stage. During each ecdysis, moulting fluids are produced and secreted into the area among the old and new cuticles. In a previous study, the protein Bombyx mori single domain von Willebrand factor type C (BmSVWC; BGIBMGA011399) was identified in the moulting fluids of Bo. mori and demonstrated to regulate ecdysis. In this study we show that in Bo. mori larvae, BmSVWC primarily locates to the integument (epidermal cells and cuticle), wing discs and head. During the moulting stage, BmSVWC is released into the moulting fluids, and is then produced again by epidermal cells after ecdysis. Fungal infection was shown to decrease the amount of BmSVWC in the cuticle, which indicates that BmSVWC is a target protein of entomopathogenic fungi. Thus, BmSVWC is mainly involved in maintaining the integrity of the integument structure, which serves to protect insects from physical damage and pathogenic infection. © 2017 The Royal Entomological Society.

  18. Bacillus thuringiensis Cry1Aa toxin-binding region of Bombyx mori aminopeptidase N.

    Science.gov (United States)

    Yaoi, K; Nakanishi, K; Kadotani, T; Imamura, M; Koizumi, N; Iwahana, H; Sato, R

    1999-12-17

    The Bacillus thuringiensis Cry1Aa toxin-binding region of Bombyx mori aminopeptidase N (APN) was analyzed, to better understand the molecular mechanism of susceptibility to the toxin and the development of resistance in insects. APN was digested with lysylendopeptidase and the ability of the resulting fragments to bind to Cry1Aa and 1Ac toxins was examined. The binding abilities of the two toxins to these fragments were different. The Cry1Aa toxin bound to the fragment containing 40-Asp to 313-Lys, suggesting that the Cry1Aa toxin-binding site is located in the region between 40-Asp and 313-Lys, while Cry1Ac toxin bound exclusively to mature APN. Next, recombinant APN of various lengths was expressed in Escherichia coli cells and its ability to bind to Cry1Aa toxin was examined. The results localized the Cry1Aa toxin binding to the region between 135-Ile and 198-Pro.

  19. Vitellogenin from the silkworm, Bombyx mori: an effective anti-bacterial agent.

    Directory of Open Access Journals (Sweden)

    Nitin Kumar Singh

    Full Text Available Silkworm, Bombyx mori, vitellogenin (Vg was isolated from perivisceral fat body of day 3 of pupa. Both Vg subunits were co-purified as verified by mass spectrometry and immunoblot. Purified Vg responded to specific tests for major posttranslational modifications on native gels indicating its nature as lipo-glyco-phosphoprotein. The Vg fraction had strong antibacterial activity against Gram negative bacterium Escherichia coli and Gram positive bacterium Bacillus subtilis. Microscopic images showed binding of Vg to bacterial cells and their destruction. When infected silkworm larvae were treated with purified Vg they survived the full life cycle in contrast to untreated animals. This result showed that Vg has the ability to inhibit the proliferation of bacteria in the silkworm fluid system without disturbing the regular metabolism of the host.

  20. The complete mitochondrial genome of Bombyx mori strain Baiyun (Lepidoptera: Bombycidae).

    Science.gov (United States)

    Zhang, Huixian; Li, Fengbo; Zhu, Xinrong; Meng, Zhiqi

    2016-05-01

    The complete mitochondrial genome of Bombyx mori strain Baiyun (Lepidoptera: Bombycidae) was determined in this study. The genome was 15,629 bp long with 37 typical animal mitochondrial genes and 1 non-coding A + T-rich region. Its gene content and order were identical to those of other lepidopteran mitochondrial genomes. All protein-coding genes (PCGs) were initiated by ATN codons except for the COI gene, which began with CGA codon. Eleven PCGs stopped with termination codon TAA, whereas the COI and COII genes ended with single T. All the tRNA genes showed typical secondary cloverleaf structures. The 496 bp AT-rich region contains several features common to other lepidopterans, such as the motif ATAGA followed by an 18-bp poly-T stretch and two microsatellite-like (TA)8 and (AT)9 elements preceded by the ATTTA motif.

  1. Genetic Divergence, Implication of Diversity, and Conservation of Silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Bharat Bhusan Bindroo

    2014-01-01

    Full Text Available Genetic diversity is critical to success in any crop breeding and it provides information about the quantum of genetic divergence and serves a platform for specific breeding objectives. It is one of the three forms of biodiversity recognized by the World Conservation Union (IUCN as deserving conservation. Silkworm Bombyx mori, an economically important insect, reported to be domesticated over 5000 years ago by human to meet his requirements. Genetic diversity is a particular concern because greater genetic uniformity in silkworm can increase vulnerability to pests and diseases. Hence, maintenance of genetic diversity is a fundamental component in long-term management strategies for genetic improvement of silkworm which is cultivated by millions of people around the worlds for its lusture silk. In this paper genetic diversity studies carried out in silkworm using divergent methods (quantitative traits and biochemical and molecular markers and present level of diversity and factors responsible for loss of diversity are discussed.

  2. Silkworm Gut Fiber of Bombyx mori as an Implantable and Biocompatible Light-Diffusing Fiber

    Directory of Open Access Journals (Sweden)

    Jose Luis Cenis

    2016-07-01

    Full Text Available This work describes a new approach to the delivery of light in deeper tissues, through a silk filament that is implantable, biocompatible, and biodegradable. In the present work, silkworm gut fibers (SGFs of Bombyx mori L., are made by stretching the silk glands. Morphological, structural, and optical properties of the fibers have been characterized and the stimulatory effect of red laser light diffused from the fiber was assayed in fibroblast cultures. SGFs are formed by silk fibroin (SF mainly in a β-sheet conformation, a stable and non-soluble state in water or biological fluids. The fibers showed a high degree of transparency to visible and infrared radiation. Using a red laser (λ = 650 nm as source, the light was efficiently diffused along the fiber wall, promoting a significant increment in the cell metabolism 5 h after the irradiation. SGFs have shown their excellent properties as light-diffusing optical fibers with a stimulatory effect on cells.

  3. Poisoning symptoms of silkworm larvae, Bombyx mori L. , by the oral application of fluorine compound

    Energy Technology Data Exchange (ETDEWEB)

    Fujii, M.

    1974-04-01

    Physiological damages caused by the oral administration of fluorine in Bombyx mori were investigated. Results obtained are summarized as follows: The alimentary canal of the larvae damaged by the fluorine became light green or light brown containing few mulberry leaves and the larvae were light brown. The growth and molting of the larvae were delayed. When larvae were affected by the fluorine, the increasing rate in body weight declined at first, followed by a reduction in the amount of food ingested, and then their development was prolonged. In chemical analyses, ingested fluorine was mostly retained in alimentary canal, digestive juice and blood, and was seldom found in the integument and the silkgland. From these results, it may be presumed that the death of silkworm larvae, caused by the oral administration of fluorine, can be mainly attributed to physiological damages of the alimentary canal caused by the fluorine toxicity, because a large amount of fluorine is accumulated in the wall of the alimentary canal.

  4. Identification of small RNA in polyhedra of Bombyx mori nuclear polyhedrosis virus

    Directory of Open Access Journals (Sweden)

    T. V. Shirina

    2014-04-01

    Full Text Available It has been shown by bioinformatic methods­ that regions of the Bombyx mori viral nuclear poly­hedrosis genome encoded two small RNA – snc RNA-1 and snc RNA-2, which could perform a structural function in polyhedra crystals formation. The aim of this work was identification of the nucleo­tide sequence of small non-coding RNAs, predicted by bioinformatic methods in B. mori polyhedra. The following methods have been used: polymera­se chain reaction, agarose gel electrophoresis, the cloning­ of PCR products, sequencing. There were first determined nucleotide sequences of snc RNA-1 and snc RNA-2 of polyhedrin mRNA complementary regions which are included in B. mori polyhedra. These RNAs have 100% identity with bioinformatic predicted sequences. These results confirmed our bioinformatic approach to the search for small RNAs encoded in B. mori nuclear polyhedrosis virus genome.

  5. Standard method for detecting Bombyx mori nucleopolyhedrovirus disease-resistant silkworm varieties

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    Yang Qiong

    Full Text Available ABSTRACT Bombyx mori nucleopolyhedrovirus (BmNPV disease is one of the most serious silkworm diseases, and it has caused great economic losses to the sericulture industry. So far, the disease has not been controlled effectively by therapeutic agents. Breeding resistant silkworm varieties breeding may be an effective way to improve resistance to BmNPV and reduce economic losses. A precise resistance-detection method will help to accelerate the breeding process. For this purpose, here we described the individual inoculation method (IIM. Details of the IIM include pathogen BmNPV preparation, mulberry leaf size, pathogen volume, rearing conditions, course of infection, and breeding conditions. Finally, a resistance comparison experiment was performed using the IIM and the traditional group inoculation method (GIM. The incidence of BmNPV infection and the within-group variance results showed that the IIM was more precise and reliable than the GIM.

  6. Transgene-based, female-specific lethality system for genetic sexing of the silkworm, Bombyx mori.

    Science.gov (United States)

    Tan, Anjiang; Fu, Guoliang; Jin, Li; Guo, Qiuhong; Li, Zhiqian; Niu, Baolong; Meng, Zhiqi; Morrison, Neil I; Alphey, Luke; Huang, Yongping

    2013-04-23

    Transgene-based genetic sexing methods are being developed for insects of agricultural and public health importance. Male-only rearing has long been sought in sericulture because males show superior economic characteristics, such as better fitness, lower food consumption, and higher silk yield. Here we report the establishment of a transgene-based genetic sexing system for the silkworm, Bombyx mori. We developed a construct in which a positive feedback loop regulated by sex-specific alternative splicing leads to high-level expression of the tetracycline-repressible transactivator in females only. Transgenic animals show female-specific lethality during embryonic and early larval stages, leading to male-only cocoons. This transgene-based female-specific lethal system not only has wide application in sericulture, but also has great potential in lepidopteran pest control.

  7. Resistance to Bombyx mori nucleopolyhedrovirus via overexpression of an endogenous antiviral gene in transgenic silkworms.

    Science.gov (United States)

    Jiang, Liang; Wang, Genhong; Cheng, Tingcai; Yang, Qiong; Jin, Shengkai; Lu, Gai; Wu, Fuquan; Xiao, Yang; Xu, Hanfu; Xia, Qingyou

    2012-07-01

    Transgenic technology is a powerful tool for improving disease-resistant species. Bmlipase-1, purified from the midgut juice of Bombyx mori, showed strong antiviral activity against B. mori nucleopolyhedrovirus (BmNPV). In an attempt to create an antiviral silkworm strain for sericulture, a transgenic vector overexpressing the Bmlipase-1 gene was constructed under the control of a baculoviral immediate early-1 (IE1) promoter. Transgenic lines were generated via embryo microinjection. The mRNA level of Bmlipase-1 in the midguts of the transgenic line was 27.3 % higher than that of the non-transgenic line. After feeding the silkworm with different amounts of BmNPV, the mortality of the transgenic line decreased to approximately 33 % compared with the non-transgenic line when the virus dose was 10(6) OB/larva. These results imply that overexpressing endogenous antiviral genes can enhance the antiviral resistance of silkworms.

  8. End-sequencing and characterization of silkworm (Bombyx mori bacterial artificial chromosome libraries

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    Narukawa Junko

    2007-09-01

    Full Text Available Abstract Background We performed large-scale bacterial artificial chromosome (BAC end-sequencing of two BAC libraries (an EcoRI- and a BamHI-digested library and conducted an in silico analysis to characterize the obtained sequence data, to make them a useful resource for genomic research on the silkworm (Bombyx mori. Results More than 94000 BAC end sequences (BESs, comprising more than 55 Mbp and covering about 10.4% of the silkworm genome, were sequenced. Repeat-sequence analysis with known repeat sequences indicated that the long interspersed nuclear elements (LINEs were abundant in BamHI BESs, whereas DNA-type elements were abundant in EcoRI BESs. Repeat-sequence analysis revealed that the abundance of LINEs might be due to a GC bias of the restriction sites and that the GC content of silkworm LINEs was higher than that of mammalian LINEs. In a BLAST-based sequence analysis of the BESs against two available whole-genome shotgun sequence data sets, more than 70% of the BESs had a BLAST hit with an identity of ≥ 99%. About 14% of EcoRI BESs and about 8% of BamHI BESs were paired-end clones with unique sequences at both ends. Cluster analysis of the BESs clarified the proportion of BESs containing protein-coding regions. Conclusion As a result of this characterization, the identified BESs will be a valuable resource for genomic research on Bombyx mori, for example, as a base for construction of a BAC-based physical map. The use of multiple complementary BAC libraries constructed with different restriction enzymes also makes the BESs a more valuable genomic resource. The GenBank accession numbers of the obtained end sequences are DE283657–DE378560.

  9. Characterisation of a desmosterol reductase involved in phytosterol dealkylation in the silkworm, Bombyx mori.

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    Leonora F Ciufo

    Full Text Available Most species of invertebrate animals cannot synthesise sterols de novo and many that feed on plants dealkylate phytosterols (mostly C(29 and C(28 yielding cholesterol (C(27. The final step of this dealkylation pathway involves desmosterol reductase (DHCR24-catalysed reduction of desmosterol to cholesterol. We now report the molecular characterisation in the silkworm, Bombyx mori, of such a desmosterol reductase involved in production of cholesterol from phytosterol, rather than in de novo synthesis of cholesterol. Phylogenomic analysis of putative desmosterol reductases revealed the occurrence of various clades that allowed for the identification of a strong reductase candidate gene in Bombyx mori (BGIBMGA 005735. Following PCR-based cloning of the cDNA (1.6 kb and its heterologous expression in Saccharomyces cerevisae, the recombinant protein catalysed reduction of desmosterol to cholesterol in an NADH- and FAD-dependent reaction.Conceptual translation of the cDNA, that encodes a 58.9 kDa protein, and database searching, revealed that the enzyme belongs to an FAD-dependent oxidoreductase family. Western blotting revealed reductase protein expression exclusively in the microsomal subcellular fraction and primarily in the gut. The protein is peripherally associated with microsomal membranes. 2D-native gel and PAGE analysis revealed that the reductase is part of a large complex with molecular weight approximately 250 kDa. The protein occurs in midgut microsomes at a fairly constant level throughout development in the last two instars, but is drastically reduced during the wandering stage in preparation for metamorphosis. Putative Broad Complex transcription factor-binding sites detectable upstream of the DHCR24 gene may play a role in this down-regulation.

  10. Isolation of a cDNA encoding a CHH-family peptide from the silkworm Bombyx mori.

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    Endo, H; Nagasawa, H; Watanabe, T

    2000-05-01

    The crustacean hyperglycemic hormone (CHH) peptide family includes four types of neuropeptide in decapod and isopod crustaceans, and the ion-transport peptide in orthopteran insects. To identify a new member of this family in Insecta, a PCR-based search for cDNAs encoding CHH-family peptides was carried out in the silkworm Bombyx mori. A cDNA, named BmCHHL (Bombyx mori CHH-like protein), with an open reading frame of 110 amino acids was isolated. Sequence analyses suggested that the conceptual protein was a precursor of a peptide of 72 amino acids which was amidated at the carboxy terminus. The BmCHHL sequence exhibited significant similarities to members of the CHH family including the orthopteran ion-transport peptide. BmCHHL expression was detected in five or six cells (per hemisphere) in the frontal area of the brain in day 4 fifth instar larvae.

  11. Genome-Wide Analysis of Host Responses to Four Different Types of Microorganisms in Bombyx Mori (Lepidoptera: Bombycidae)

    OpenAIRE

    Cheng, Tingcai; Lin, Ping; Huang, Lulin; Wu, Yuqian; Jin, Shengkai; Liu, Chun; Xia, Qingyou

    2016-01-01

    Several pathogenic microorganisms have been used to investigate the genome-wide transcriptional responses of Bombyx mori to infection. However, studies have so far each focused on one microorganism, and systematic genome-wide comparison of transcriptional responses to different pathogenic microorganisms has not been undertaken. Here, we surveyed transcriptional responses of B. mori to its natural bacterial, viral, and fungal pathogens, Bacillus bombyseptieus, B. mori nucleopolyhedrovirus (BmN...

  12. Release of Ecdysteroid-Phosphates from Egg Yolk Granules and Their Dephosphorylation during Early Embryonic Development in Silkworm, Bombyx mori

    OpenAIRE

    Yamada, Ryouichi; Yamahama, Yumi; Sonobe, Haruyuki

    2005-01-01

    Newly laid eggs of many insect species store maternal ecdysteroids as physiologically inactive phosphoric esters. In the silkworm Bombyx mori, we previously reported the presence of a specific enzyme, called ecdysteroid-phosphate phosphatase (EPPase), which catalyzes the dephosphorylation of ecdysteroid-phosphates to increase the amount of free ecdysteroids during early embryonic development. In this study, we demonstrated that (1) EPPase is found in the cytosol of yolk cells, (2) ecdysteroid...

  13. Genome-Wide Identification of Polycomb Target Genes Reveals a Functional Association of Pho with Scm in Bombyx mori

    OpenAIRE

    Li, Zhiqing; Cheng, Daojun; Mon, Hiroaki; Tatsuke, Tsuneyuki; Zhu, Li; Xu, Jian; Lee, Jae Man; Xia, Qingyou; Kusakabe, Takahiro

    2012-01-01

    Polycomb group (PcG) proteins are evolutionarily conserved chromatin modifiers and act together in three multimeric complexes, Polycomb repressive complex 1 (PRC1), Polycomb repressive complex 2 (PRC2), and Pleiohomeotic repressive complex (PhoRC), to repress transcription of the target genes. Here, we identified Polycomb target genes in Bombyx mori with holocentric centromere using genome-wide expression screening based on the knockdown of BmSCE, BmESC, BmPHO, or BmSCM gene, which represent ...

  14. Modulation of gene expression by nutritional state and hormones in Bombyx larvae in relation to its growth period.

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    Thounaojam, Bembem; Keshan, Bela

    2017-11-01

    Insect growth and development are mainly regulated via synchronization of many extrinsic and intrinsic factors such as nutrition and hormones. Previously we have demonstrated that larval growth period influences the effect of insulin on the accumulation of glycogen in the fat body of Bombyx larvae. In the present study we demonstrate that Bombyx larvae at the terminal growth period (TGP, after critical weight) had a significantly greater increase in the expression level of Akt in the fat body than at the active growth period (AGP, before critical weight). The larvae at TGP also showed an increase in the expression level of ecdysone receptors (EcRB1 and USP1) and ecdysone-induced early genes (E75A and broad). The treatment of bovine insulin and methoprene to larvae at AGP induced the transcript levels of Akt, irrespective of the nutritional status of the larvae. However, in larvae at TGP, insulin repressed the transcript level of Akt. On contrary, 20-hydroxyecdysone (20E) induced the expression level of Akt in TGP larvae, but at feeding only. Insulin and 20E thus showed an antagonistic action on the Akt expression level in TGP larvae under feeding. The studies thus showed that larval growth period influences the expression level of Akt and ecdysone receptors in Bombyx. Further, the growth period and nutrition modulate the effect of exogenous hormones on Akt expression. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Baculovirus LEF-11 Hijack Host ATPase ATAD3A to Promote Virus Multiplication in Bombyx mori cells.

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    Dong, Zhan-Qi; Hu, Nan; Dong, Fei-Fan; Chen, Ting-Ting; Jiang, Ya-Ming; Chen, Peng; Lu, Cheng; Pan, Min-Hui

    2017-04-10

    Research on molecular mechanisms that viruses use to regulate the host apparatus is important in virus infection control and antiviral therapy exploration. Our previous research showed that the Bombyx mori nucleopolyhedrovirus (BmNPV) LEF-11 localized to dense regions of the cell nucleus and is required for viral DNA replication. Herein, we examined the mechanism of LEF-11 on BmNPV multiplication and demonstrated that baculovirus LEF-11 interacts with Bombyx mori ATAD3A and HSPD1 (HSP60) protein. Furthermore, we showed that LEF-11 has the ability to induce and up-regulate the expression of ATAD3A and HSPD1, phenomena that were both reversed upon knockdown of lef-11. Our findings showed that ATAD3A and HSPD1 were necessary and contributed to BmNPV multiplication in Bombyx mori cells. Moreover, ATAD3A was found to directly interact with HSPD1. Interestingly, ATAD3A was required for the expression of HSPD1, while the knockdown of HSPD1 had no obvious effect on the expression level of ATAD3A. Taken together, the data presented in the current study demonstrated that baculovirus LEF-11 hijacks the host ATPase family members, ATAD3A and HSPD1, efficiently promote the multiplication of the virus. This study furthers our understanding of how baculovirus modulates energy metabolism of the host and provides a new insight into the molecular mechanisms of antiviral research.

  16. Defense role of the cocoon in the silk worm Bombyx mori L.

    Science.gov (United States)

    Pandiarajan, Jeyaraj; Cathrin, Britto P; Pratheep, Thangaraj; Krishnan, Muthukalingan

    2011-11-15

    Silk from the domesticated silk worm Bombyx mori procures foreign body response naturally, so it has been utilized as a biomaterial for decades. In India the prime focus of the sericulture industry is to improve silk production with high quality silk. Naturally, the silk worm builds its cocoon not only with silk proteins, but also with antimicrobial proteins to avoid infection since the cocoon is non-motile and non-feeding. The aim of the present study is to elucidate the antimicrobial proteins that persist in the cocoon of the silk worm Bombyx mori. At the pupal stage, the silk worm cocoon shell extract was prepared from the day of pupation (P0) to the day of natural rupture of the cocoon for the eclosion of moth (NR). Using the cocoon shell extract a microbial susceptibility test was performed by the disc diffusion method against the microbes Escherchia coli, Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, and Klebsiella pneumoniae. The development of a zone of inhibition against the microbes confirmed the presence of antimicrobial/immunogenic activity of the cocoon shell extract. For further analysis, the cocoon shell extract was subjected to 7-15% sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE). The protein profile of the cocoon extract revealed the coomassie blue stained bands resolved from the 150-15 kDa molecular range. Interestingly, a polypeptide localized at around 29 kDa showed remarkable expressional changes during the development of pupa. To characterize the 29 kDa protein, it was eluted from the gel, digested with trypsin and analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The trypsin-digested peptide peaks were analyzed through MASCOT and peptides were matched with the NCBI nr database. The peptides were very well matched with the 18 wheeler protein, which is reported to be responsible for innate immunity, belonging to the Toll family in insects and responsible for cellular

  17. Efficient silkworm expression of human GPCR (nociceptin receptor) by a Bombyx mori bacmid DNA system

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    Kajikawa, Mizuho; Sasaki, Kaori [Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582 (Japan); Wakimoto, Yoshitaro; Toyooka, Masaru [Department of Chemistry and Chemical Biology, Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515 (Japan); Motohashi, Tomoko; Shimojima, Tsukasa [National Institute of Genetics, 1111 Yata, Mishima, Shizuoka 411-8540 (Japan); Takeda, Shigeki [Department of Chemistry and Chemical Biology, Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515 (Japan); Park, Enoch Y. [Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Oya, Suruga-ku, Shizuoka, Shizuoka 422-8529 (Japan); Maenaka, Katsumi, E-mail: kmaenaka-umin@umin.net [Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582 (Japan)

    2009-07-31

    Guanine nucleotide-binding protein (G protein) coupled receptors (GPCRs) are frequently expressed by a baculovirus expression vector system (BEVS). We recently established a novel BEVS using the bacmid system of Bombyx mori nucleopolyhedrovirus (BmNPV), which is directly applicable for protein expression in silkworms. Here, we report the first example of GPCR expression in silkworms by the simple injection of BmNPV bacmid DNA. Human nociceptin receptor, an inhibitory GPCR, and its fusion protein with inhibitory G protein alpha subunit (G{sub i}{alpha}) were both successfully expressed in the fat bodies of silkworm larvae as well as in the BmNPV viral fraction. Its yield was much higher than that from Sf9 cells. The microsomal fractions including the nociceptin receptor fusion, which are easily prepared by only centrifugation steps, exhibited [{sup 35}S]GTP{gamma}S-binding activity upon specific stimulation by nociceptin. Therefore, this rapid method is easy-to-use and has a high expression level, and thus will be an important tool for human GPCR production.

  18. Mechanical properties of cocoons constructed consecutively by a single silkworm caterpillar, Bombyx mori

    Science.gov (United States)

    Huang, S. Q.; Zhao, H. P.; Feng, X. Q.; Cui, W.; Lin, Z.; Xu, M. Q.

    2008-04-01

    Most animals have the ability to adapt, to some extends and in different ways, the variation or disturbance of environment. In our experiments, we forced a silkworm caterpillar to spin two, three or four thin cocoons by taking it out from the cocoon being constructed. The mechanical properties of these cocoons were studied by static tensile tests and dynamic mechanical thermal analysis. Though external disturbances may cause the decrease in the total weight of silk spun by the silkworm, a gradual enhancement was interestingly found in the mechanical properties of these thin cocoons. Scanning electron microscopy observations of the fractured specimens of the cocoons showed that there exist several different energy dissipation mechanisms occurred simultaneously at macro-, meso-, and micro-scales, yielding a superior capacity of cocoons to adsorb the energy of possible attacks from the outside and to protect efficiently its pupa against damage. Through evolution of millions of years, therefore, the silkworm Bombyx mori seems to have gained the ability to adapt external disturbances and to redesign a new cocoon with optimized protective function when its first cocoon has been damaged for some reasons.

  19. FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori.

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    Ding-Pei Long

    Full Text Available A comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they have not yet been established for use in the manipulation of the silkworm Bombyx mori genome. In this study, we achieved site-specific excision of a target gene at predefined chromosomal sites in the silkworm using a FLP/FRT site-specific recombination system. We first constructed two stable transgenic target silkworm strains that both contain a single copy of the transgene construct comprising a target gene expression cassette flanked by FRT sites. Using pre-blastoderm microinjection of a FLP recombinase helper expression vector, 32 G3 site-specific recombinant transgenic individuals were isolated from five of 143 broods. The average frequency of FLP recombinase-mediated site-specific excision in the two target strains genome was approximately 3.5%. This study shows that it is feasible to achieve site-specific recombination in silkworms using the FLP/FRT system. We conclude that the FLP/FRT system is a useful tool for genome manipulation in the silkworm. Furthermore, this is the first reported use of the FLP/FRT system for the genetic manipulation of a lepidopteran genome and thus provides a useful reference for the establishment of genome manipulation technologies in other lepidopteran species.

  20. PERTUMBUHAN ULAT SUTERA BOMBYX MORI (LEPIDOPTERA: BOMBYCIDAE PADA BERBAGAI SUMBER PROTEIN DALAM PAKANNYA

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    Santoso

    2013-08-01

    Full Text Available Silkworm rearing has high economical value for producing silk material and other beneficial biological materials such as enzyme, antibacterial agent, and antioxidant. For those purposes, it is important to continuously support high quality feed in the silkworm rearing.This study was aimed to evaluate study the growth of silkworm Bombyx mori fed with artificial diet containing different resources of protein (blood meal or soybean meal. The experiment used completely randomized design 5 five treatments artificial feed formulas and 20 repetitions. Each experiment unit consisted of 1 larvae that was reared individually and given different feed. Formula A: 100% blood meal (BM; B: 75% (BM + 25% soybean meal (SM; C: 50% (BM + 50% (SM; D: 25% (BM+ 75% (SM; and E: 100% SM. Parameters measured were the feed consumption/larvae, opproximate digestibility, ECI (efficiency of conversion of feed ingested, ECD (efficiency conversion of feed digestibility, and body weight gain in fourth and fifth of the larvae instar. Oxygen consumption were measured on the fifth day of the fifth instar before meal and one hour after meal. The result showed that resource of protein did not significantly affect the parameters on the fourth instar, but significantly affected on the fifth instar larvae. Increment of soybean meal in artificial diet increasing oxygen consumption, and resulted decreassing of ECI and ECD value on the fifth instar larvae.

  1. Effects of transient high temperature treatment on the intestinal flora of the silkworm Bombyx mori.

    Science.gov (United States)

    Sun, Zhenli; Kumar, Dhiraj; Cao, Guangli; Zhu, Liyuan; Liu, Bo; Zhu, Min; Liang, Zi; Kuang, Sulan; Chen, Fei; Feng, Yongjie; Hu, Xiaolong; Xue, Renyu; Gong, Chengliang

    2017-06-13

    The silkworm Bombyx mori is a poikilotherm and is therefore sensitive to various climatic conditions. The influence of temperature on the intestinal flora and the relationship between the intestinal flora and gene expression in the silkworm remain unknown. In the present study, changes of the intestinal flora at 48, 96 and 144 h following transient high temperature treatment (THTT) of 37 °C for 8 h were investigated. According to principal component analysis, the abundances of Enterococcus and Staphylococcus showed a negative correlation with other dominant genera. After THTT, the gene expression levels of spatzle-1 and dicer-2 were increased and decreased, respectively, which suggested that the Toll and RNAi pathways were activated and suppressed, respectively. The species-gene expression matrix confirmed that the spatzle-1 and dicer-2 gene expression levels were negatively and positively correlated, respectively, with the abundance of Enterococcus and Staphylococcus in the control. The abundance of Variovorax post-THTT was positively correlated with the spatzle-1 gene expression level, whereas the community richness of Enterococcus was negatively correlated with the spatzle-1 gene expression level and positively correlated with the dicer-2. The results of the present investigation provide new evidence for understanding the relationships among THTT, intestinal flora and host gene expression.

  2. Traditional Uses, Origins, Chemistry and Pharmacology of Bombyx batryticatus: A Review

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    Meibian Hu

    2017-10-01

    Full Text Available Bombyx batryticatus (B. batryticatus, a well-known traditional animal Chinese medicine, has been commonly used in China for thousands of years. The present paper reviewed advances in traditional uses, origin, chemical constituents, pharmacology and toxicity studies of B. batryticatus. The aim of the paper is to provide more comprehensive references for modern B. batryticatus study and application. In Traditional Chinese Medicine (TCM culture, drugs containing B. batryticatus have been used to treat convulsions, headaches, skin prurigo, scrofula, tonsillitis and fever. Many studies indicate B. batryticatus contains various compounds, including protein and peptides, fatty acids, flavonoids, nucleosides, steroids, coumarin, polysaccharide and others. Numerous investigations also have shown that extracts and compounds from B. batryticatus exert a wide spectrum of pharmacological effects both in vivo and in vitro, including effects on the nervous system, anticoagulant effects, antitumor effects, antibacterial and antifungal effects, antioxidant effects, hypoglycemic effects, as well as other effects. However, further studies should be undertaken to investigate bioactive compounds (especially proteins and peptides, toxic constituents, using forms and the quality evaluation and control of B. batryticatus. Furthermore, it will be interesting to study the mechanism of biological activities and structure-function relationships of bioactive constituents in B. batryticatus.

  3. COCOON PRODUCTION OF THE SILKWORM, Bombyx mori L. (LEPIDOPTERA: BOMBYCIDAE, FED ON LEAVES OF MULBERRY HYBRIDS

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    GERBSON AZEVEDO DE MENDONÇA

    2010-01-01

    Full Text Available Brazil is the fourth cocoon producer in the world. In São Paulo State there are mulberry some hybrids whose productivity are higher than the commonly cultivated varieties. The objective of this study was to evaluate the effect of mulberry hybrids (Morus spp. on the cocoon production of silkworm (Bombyx mori L.. The experiment was conducted at the Unidade Regional de Pesquisa de Gália do Instituto de Zootecnia, SP. The caterpillars were fed on leaves of the hybrids IZ-3/2, IZ-13/6, IZ-15/7, IZ-19/13, IZ-56/4, IZ-57/2, IZ- 40, IZ-64, in a rearing hut at 25 oC ± 3 oC and 75% ± 5% relative humidity. 'Korin' was used as standard. The hybrids affected the duration of the larval period and the weight of the caterpillars, prepupaes and the silk glands as well. There was a reduction in the duration of larval development when the caterpillars had been fed with hybrid IZ-56/4 and the 'Korin' variety. Hybrids IZ-57/2, IZ-56/4 and IZ-15/7 presented the highest cocoon production.

  4. Appearance of differentiated cells derived from polar body nuclei in the silkworm, Bombyx mori

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    Hiroki eSakai

    2013-09-01

    Full Text Available AbstractIn Bombyx mori, polar body nuclei are observed until 9h after egg lying, however, the fate of polar body nuclei remains unclear. To examine the fate of polar body nuclei, we employed a mutation of serosal cell pigmentation, pink-eyed white egg (pe. The heterozygous pe/+pe females produced black serosal cells in white eggs, while pe/pe females did not produce black serosal cells in white eggs. These results suggest that the appearance of black serosal cells in white eggs depends on the genotype (pe/ +pe of the mother. Because the polar body nuclei had +pe genes in the white eggs laid by a pe/ +pe female, polar body nuclei participate in development and differentiate into functional cell (serosal cells. Analyses of serosal cells pigmentation indicated that approximately 30% of the eggs contained polar-body-nucleus-derived cells. These results demonstrate that polar-body-nucleus-derived cells appeared at a high frequency under natural conditions. Approximately 80% of polar-body-nucleus-derived cells appeared near the anterior pole and the dorsal side, which is opposite to where embryogenesis occurs. The number of cells derived from the polar body nuclei was very low. Approximately 26 % of these eggs contained only one black serosal cell. PCR-based analysis revealed that the polar-body-nucleus-derived cells disappeared in late embryonic stages (stage 25. Overall, polar-body-nuclei-derived cells were unlikely to contribute to embryos.

  5. Preparation and characterization of regenerated fiber from the aqueous solution of Bombyx mori cocoon silk fibroin

    Energy Technology Data Exchange (ETDEWEB)

    Zhu Zhenghua [Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588 (Japan); Department of Application Engineering, ZheJiang Vocational College of Economic and Trade, HangZhou, ZheJiang 310018 (China); Imada, Takuzo [Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588 (Japan); Asakura, Tetsuo, E-mail: asakura@cc.tuat.ac.jp [Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588 (Japan)

    2009-10-15

    The regenerated silk fibers with high strength and high biodegradability were prepared from the aqueous solution of Bombyx mori silk fibroin from cocoons with wet spinning method. Although the tensile strength of the regenerated silk fibroin fiber, 210 MPa is still half of the strength of native silk fiber, the diameter of the fiber is about 100 {mu}m which is suitable for monofilament of suture together with high biodegradability. The high concentration (30%, w/v) of the aqueous solution of the silk fibroin which corresponds to the high concentration in the middle silkgland of silkworm was obtained. This was performed by adjusting the pH of the aqueous solution to 10.4 which corresponds to pK{sub a} value of the OH group of Tyr residues in the silk fibroin. The mixed solvent, methanol/acetic acid (7:3 in volume ratio) was used as coagulant solvent for preparing the regenerated fiber. The structural change of silk fibroin fiber by stretching was monitored with both {sup 13}C solid state NMR and X-ray diffraction methods, indicating that the high strength of the fiber is related with the long-range orientation of the silk fibroin chain with {beta}-sheet structure.

  6. Vibration receptive sensilla on the wing margins of the silkworm moth Bombyx mori.

    Science.gov (United States)

    Ai, Hiroyuki; Yoshida, Akihiro; Yokohari, Fumio

    2010-03-01

    Bristles along the wing margins (wm-bristles) of the silkworm moth, Bombyx mori, were studied morphologically and electrophysiologically. The male moth has ca. 50 wm-bristles on each forewing and hindwing. Scanning electron microscopy revealed that these wm-bristles are typical mechanosensilla. Leuco-methylene blue staining demonstrated that each wm-bristle has a single receptor neuron, which is also characteristic of the mechanosensillum. The receptor neuron responded to vibrating air currents but did not respond to a constant air current. The wm-bristles showed clear directional sensitivity to vibrating air currents. The wm-bristles were classified into two types, type I and type II, by their response patterns to sinusoidal movements of the bristle. The neuron in type I discharged bursting spikes immediately following stimulation onset and also discharged a single spike for each sinusoidal cycle for frequencies less than ca. 60 Hz. The neuron in type II only responded to vibrations over 40 Hz and, specifically at 75 Hz, discharged a single spike for each sinusoidal cycle throughout the stimulation period. These results suggest that the two types of wm-bristles are highly tuned in different ways to detect vibrations due to the wing beat. The roles of the wm-bristles in the wing beat are discussed. Copyright (c) 2009 Elsevier Ltd. All rights reserved.

  7. Immobilization of foreign protein into polyhedra of Bombyx mori nucleopolyhedrovirus (BmNPV).

    Science.gov (United States)

    Xiang, Xing-wei; Yang, Rui; Chen, Lin; Hu, Xiao-long; Yu, Shao-fang; Cao, Cui-ping; Wu, Xiao-feng

    2012-02-01

    In the late phase of Bombyx mori nucleopolyhedrovirus (BmNPV) infection, a large amount of polyhedra appear in the infected cell nucleolus, these polyhedra being dense protein crystals protecting the incorporated virions from the harsh environment. To investigate whether the foreign protein could be immobilized into the polyhedra of BmNPV, two recombinant baculoviruses were generated by a novel BmNPV polyhedrin-plus (polh(+)) Bac-to-Bac system, designated as vBmBac(polh(+))-enhanced green fluorescent protein (EGFP) and vBmBac(polh(+))-LacZ, which can express the polyhedrin and foreign protein simultaneously. Light microscopy analysis showed that all viruses produced polyhedra of normal appearance. Green fluorescence can be apparently detected on the surface of the vBmBac(polh(+))-EGFP polyhedra, but not the BmNPV polyhedra. Fluorescence analysis and anti-desiccation testing confirmed that EGFP was embedded in the polyhedra. As expected, the vBmBac(polh(+))-LacZ polyhedra contained an amount of LacZ and had a higher β-galactosidase activity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were also performed to verify if the foreign proteins were immobilized into polyhedra. This study provides a new inspiration for efficient preservation of useful proteins and development of new pesticides with toxic proteins.

  8. Immobilization of foreign protein into polyhedra of Bombyx mori nucleopolyhedrovirus (BmNPV)*

    Science.gov (United States)

    Xiang, Xing-wei; Yang, Rui; Chen, Lin; Hu, Xiao-long; Yu, Shao-fang; Cao, Cui-ping; Wu, Xiao-feng

    2012-01-01

    In the late phase of Bombyx mori nucleopolyhedrovirus (BmNPV) infection, a large amount of polyhedra appear in the infected cell nucleolus, these polyhedra being dense protein crystals protecting the incorporated virions from the harsh environment. To investigate whether the foreign protein could be immobilized into the polyhedra of BmNPV, two recombinant baculoviruses were generated by a novel BmNPV polyhedrin-plus (polh+) Bac-to-Bac system, designated as vBmBac(polh+)-enhanced green fluorescent protein (EGFP) and vBmBac(polh+)-LacZ, which can express the polyhedrin and foreign protein simultaneously. Light microscopy analysis showed that all viruses produced polyhedra of normal appearance. Green fluorescence can be apparently detected on the surface of the vBmBac(polh+)-EGFP polyhedra, but not the BmNPV polyhedra. Fluorescence analysis and anti-desiccation testing confirmed that EGFP was embedded in the polyhedra. As expected, the vBmBac(polh+)-LacZ polyhedra contained an amount of LacZ and had a higher β-galactosidase activity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were also performed to verify if the foreign proteins were immobilized into polyhedra. This study provides a new inspiration for efficient preservation of useful proteins and development of new pesticides with toxic proteins. PMID:22302424

  9. [Mutagenic analysis on the polyhedrin gene (polh) of Bombyx mori nuclear polyhedrosis virus (BmNPV)].

    Science.gov (United States)

    Cao, Y; Xiao, Q X; Huang, Y D; Ge, C B; Huang, Z R; Liu, L S

    2000-01-01

    In our early studies, the abnormal shape of the polyhedra of Bombyx mori nuclear polyhedrosis virus (BmNPV) induced by chemical mutagens of MMC. 9-AA and EMS occurred, and the genome of the mutated BmNPV obtained from the successive test had some change in the restriction endonuclease partners of EcoRI, BglII and BamHI. The present studies showed that the arrangement of the crystal lattice of the polyhedrin was disorderly, and the SDS-PAGE electropherogram of the polyhedrin depicted distinct change in comparison with control group. The results of sequencing analysis showed that many point mutations with characteristics of the base substitution had occurred at some sites of the BmNPV polh gene in three mutated groups, and these results funther revealed molecular mutagenesis of the mutagens effective to BmNPV. It was not confirmable that the point mutations of polh gene in the mutated BmNPV have relationship to abnormal shape of the polyhedra.

  10. Nonvirus encoded proteins could be embedded into Bombyx mori cypovirus polyhedra.

    Science.gov (United States)

    Zhang, Yi-Ling; Xue, Ren-Yu; Cao, Guang-Li; Meng, Xiang-Kun; Zhu, Yue-Xiong; Pan, Zhong-Hua; Gong, Cheng-Liang

    2014-01-01

    To explore whether the nonvirus encoded protein could be embedded into Bombyx mori cypovirus (BmCPV) polyhedra. The stable transformants of BmN cells expressing a polyhedrin (Polh) gene of BmCPV were constructed by transfection with a non-transposon derived vector containing a polh gene. The polyhedra were purified from the midguts of BmCPV-infected silkworms and the transformed BmN cells, respectively. The proteins embedded into polyhedra were determined by mass spectrometry analysis. Host derived proteins were detected in the purified polyhedra. Analysis of structure and hydrophilicity of embedded proteins indicated that the hydrophilic proteins, in structure, were similar to the left-handed structure of polyhedrin or the N-terminal domain of BmCPV structural protein VP3, which were easily embedded into the BmCPV polyhedra. The lysate of polyhedra purified from the infected transformation of BmN cells with modified B. mori baculovirus BmPAK6 could infect BmN cells, indicating that B. mori baculovirus could be embedded into BmCPV polyhedra. Both the purified polyhedra and its lysate could be coloured by X-gal, indicating that the β-galactosidase expressed by BmPAK6 could be incorporated into BmCPV polyhedra. These results suggested that some heterologous proteins and baculovirus could be embedded into polyhedra in an unknown manner.

  11. Physicochemical Properties of Meat Batter Added with Edible Silkworm Pupae (Bombyx mori) and Transglutaminase.

    Science.gov (United States)

    Park, Yoo-Sun; Choi, Yun-Sang; Hwang, Ko-Eun; Kim, Tae-Kyung; Lee, Cheol-Won; Shin, Dong-Min; Han, Sung Gu

    2017-01-01

    This study was conducted to investigate the physicochemical properties of meat batters prepared with fresh pork meat, back fat, water, and salt and formulated with three different amounts (5%, 10%, and 15%) of silkworm pupae ( Bombyx mori ) powder and transglutaminase (TG). Meat batters formulated with silkworm pupae powder showed significantly higher contents of protein and ash than control batter. Addition of silkworm pupae to batter also showed significantly lower cooking loss than the control. Moreover, meat batter containing 15% silkworm pupae showed no significant difference in redness value compared to the control. In addition, pH, viscosity, hardness, gumminess, and chewiness were improved after the addition of silkworm pupae. Furthermore, meat batter formulated with TG and silkworm pupae showed improved hardness, gumminess, chewiness and viscosity compared to control batter. Addition of 1% TG with 15% silkworm pupae to meat batter resulted in significantly higher pH, textures, and viscosity. Our data suggest that both silkworm pupae and TG can be added to meat batter to improve its physicochemical properties. Therefore, combination of silkworm pupae and TG could be a new nutritional and functional source for meat products.

  12. Expression of RYamide in the nervous and endocrine system of Bombyx mori.

    Science.gov (United States)

    Roller, Ladislav; Čižmár, Daniel; Bednár, Branislav; Žitňan, Dušan

    2016-06-01

    RYamides are neuropeptides encoded by a gene whose precise expression and function have not yet been determined. We identified the RYamide gene transcript (fmgV1g15f, SilkBase database) and predicted two candidates for G-protein coupled RYamide receptors (A19-BAG68418 and A22-BAG68421) in the silkworm Bombyx mori. We cloned the RYamide transcript and described its spatial expression using in situ hybridisation. In the larval central nervous system (CNS) expression of RYamide was restricted to 12-14 small neurons in the brain and two posterior neurons in the terminal abdominal ganglion. During metamorphosis their number decreased to eight protocerebral neurons in the adults. Multiple staining, using various insect neuropeptide antibodies, revealed that neurons expressing RYamide are different from other peptidergic cells in the CNS. We also found RYamide expression in the enteroendocrine cells (EC) of the anterior midgut of larvae, pupae and adults. Two minor subpopulations of these EC were also immunoreactive to antibodies against tachykinin and myosupressin. This expression pattern suggests RYamides may play a role in the regulation of feeding and digestion. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. CRISPR disruption of TCTP gene impaired normal development in the silkworm Bombyx mori.

    Science.gov (United States)

    Liu, Zu-Lian; Xu, Jun; Ling, Lin; Zhang, Ru; Shang, Peng; Huang, Yong-Ping

    2018-01-09

    The translationally controlled tumor protein (TCTP) is a highly conserved and multifunctional protein with activities ranging from cytoskeletal regulation to transcription regulation in numerous organisms. In insects, TCTP is essential for cell growth and proliferation. Recently, TCTP has been reported to affect the innate intestinal immune pathway in the Bombyx mori silkworm, a lepidopteran model insect. However, the comprehensive physiological roles of TCTP in the silkworm remain poorly understood. Here, we performed functional analysis of BmTCTP by using a binary transgenic CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/RNA-guided CRISPER-associated protein 9 nucleases) system. Disruption of BmTCTP led to developmental arrestment and subsequent lethality in third instar larvae. Histological analysis revealed that growth impairment originated from decreased cell size, and the proliferation and differentiation of intestinal epithelial cells were also affected. RNA-seq analysis revealed that genes involved in carbohydrate metabolism, lipid metabolism and digestive system pathways were significantly affected by BmTCTP depletion. Together, the results demonstrated that BmTCTP plays a key role in controlling larval growth and development. © 2018 Institute of Zoology, Chinese Academy of Sciences.

  14. Processing and characterization of silk sericin from Bombyx mori and its application in biomaterials and biomedicines.

    Science.gov (United States)

    Cao, Ting-Ting; Zhang, Yu-Qing

    2016-04-01

    Bombyx mori silk is composed of 60-80% fibroin, 15-35% sericin and 1-5% non-sericin component including wax, pigments, sugars and other impurities. For two decades, the protein-based silk fibroin was extensively used in the research and development of medical biomaterials and biomedicines. Sericin is frequently ignored and abandoned as a byproduct or waste in the processing of traditional silk fabrics, silk floss or modern silk biomaterials. However, similar to fibroin, sericin is not only a highly useful biological material, but also a lot of biological activity. Moreover, the non-sericin component present with sericin in the cocoon shell also has a strong biological activity. In this review, the extraction and recovery methods of sericin and the non-sericin component from the cocoon layer are reported, and their composition, properties and biological activity are described to produce a comprehensive report on biomedical materials and biological drugs. In addition, related problems or concerns present in the research and development of sericin are discussed, and a potential application of sericin in sustainable development is also presented. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Enhancing effect of glycerol on the tensile properties of Bombyx mori cocoon sericin films.

    Science.gov (United States)

    Zhang, Haiping; Deng, Lianxia; Yang, Mingying; Min, Sijia; Yang, Lei; Zhu, Liangjun

    2011-01-01

    An environmental physical method described herein was developed to improve the tensile properties of Bombyx mori cocoon sericin films, by using the plasticizer of glycerol, which has a nontoxic effect compared with other chemical crosslinkers. The changes in the tensile characteristics and the structure of glycerolated (0-40 wt% of glycerol) sericin films were investigated. Sericin films, both in dry and wet states, showed enhanced tensile properties, which might be regulated by the addition of different concentrations of glycerol. The introduction of glycerol results in the higher amorphous structure in sericin films as evidenced by analysis of attenuated total reflection Fourier transform infrared (ATR-FTIR) spectra, thermogravimetry (TGA) and differential scanning calorimetry (DSC) curves. Scanning Electron Microscopy (SEM) observation revealed that glycerol was homogeneously blended with sericin molecules when its content was 10 wt%, while a small amount of redundant glycerol emerged on the surface of sericin films when its content was increased to 20 wt% or higher. Our results suggest that the introduction of glycerol is a novel nontoxic strategy which can improve the mechanical features of sericin-based materials and subsequently promote the feasibility of its application in tissue engineering.

  16. Enhancing Effect of Glycerol on the Tensile Properties of Bombyx mori Cocoon Sericin Films

    Directory of Open Access Journals (Sweden)

    Liangjun Zhu

    2011-05-01

    Full Text Available An environmental physical method described herein was developed to improve the tensile properties of Bombyx mori cocoon sericin films, by using the plasticizer of glycerol, which has a nontoxic effect compared with other chemical crosslinkers. The changes in the tensile characteristics and the structure of glycerolated (0–40 wt% of glycerol sericin films were investigated. Sericin films, both in dry and wet states, showed enhanced tensile properties, which might be regulated by the addition of different concentrations of glycerol. The introduction of glycerol results in the higher amorphous structure in sericin films as evidenced by analysis of attenuated total reflection Fourier transform infrared (ATR-FTIR spectra, thermogravimetry (TGA and differential scanning calorimetry (DSC curves. Scanning Electron Microscopy (SEM observation revealed that glycerol was homogeneously blended with sericin molecules when its content was 10 wt%, while a small amount of redundant glycerol emerged on the surface of sericin films when its content was increased to 20 wt% or higher. Our results suggest that the introduction of glycerol is a novel nontoxic strategy which can improve the mechanical features of sericin-based materials and subsequently promote the feasibility of its application in tissue engineering.

  17. An active recombinant cocoonase from the silkworm Bombyx mori: bleaching, degumming and sericin degrading activities.

    Science.gov (United States)

    Unajak, Sasimanas; Aroonluke, Suradet; Promboon, Amornrat

    2015-04-01

    Cocoonase is a serine protease produced by silk moths and used for softening the cocoons so that they can escape. Degumming is one of the important steps in silk processing. This research aimed to produce an active recombinant Bombyx mori cocoonase (BmCoc) for the silk degumming process. A recombinant BmCoc was successfully expressed in a Pichia pastoris system. The purified enzyme showed specific activity of 227 U mg(-1) protein, 2.4-fold purification, 95% yield and a molecular weight of 26 kDa. The enzyme exhibited optimal temperature at 40 °C and optimal pH at 8, and showed thermal stability at 25-45 °C and pH stability at 5-9. The recombinant enzyme exhibited sericin degumming ability and color bleaching characteristics, and did not affect the fibroin fiber. The enzyme also degraded sericin substrate with a product size about 30-70 kDa. In this study, we successfully produced the active recombinant BmCoc in P. pastoris with promising functions for the Thai silk degumming process, which includes degumming, sericin degrading and color bleaching activities. Our data clearly indicated that the recombinant enzyme had proteolytic activity on sericin but not on fibroin proteins. The recombinant BmCoc has proven to be suitable for numerous applications in the silk industry. © 2014 Society of Chemical Industry.

  18. Melanin pigmentation gives rise to black spots on the wings of the silkworm Bombyx mori.

    Science.gov (United States)

    Ito, Katsuhiko; Yoshikawa, Manabu; Fujii, Takeshi; Tabunoki, Hiroko; Yokoyama, Takeshi

    2016-01-01

    Several mutants of the silkworm Bombyx mori show body color variation at the larval and adult stages. The Wild wing spot (Ws) mutant exhibits a phenotype in which the moth has a spot on the apex of the forewing. In this study, we investigated this trait to elucidate the molecular mechanism underlying the color pattern. Microscopy of the black spot of Ws mutants showed that the pigment emerges in the scales of the wing, and accumulation of the pigment becomes strong just before eclosion. We next examined the relationship between the black spot of the Ws mutant and melanin. The spectrophotometry using alkaline extracts from the black spot in the wing showed the highest absorption intensity at 405nm, which is the absorbance wavelength of melanin. Moreover, inhibition assays for enzymes implicated in melanin synthesis using 3-iodo-l-tyrosine (a tyrosine hydroxylase inhibitor) and L-α-methyl-DOPA (a dopa decarboxylase inhibitor) revealed that treatment with each inhibitor disrupted the pigmentation of the wing of the Ws mutant. On the basis of these results, we analyzed the expression pattern of five genes involved in melanin formation, and found that the expression levels of yellow and laccase2 were increased just before pigmentation, whereas those of DDC, tan, and TH were increased when the apex of the wing turned black. These results showed that melanin pigmentation gives rise to the black spot on the wing. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Silkworm (Bombyx mori) hemolymph unable to substitute fetal bovine serum in insect cell culture

    Science.gov (United States)

    Suparto, Irma H.; Khalam, Chandra Nur; Praira, Willy; Sajuthi, Dondin

    2014-03-01

    Fetal Bovine Serum (FBS) in animal cell culture media is an important source of nutrients for cell growth. However, the harvest and collection of FBS cause bioethical concerns. Efforts to reduce and preferably replace FBS with synthetic or other natural alternatives are continually being explored. Hemolymph silkworm (Bombyx mori) contains many nutrients needed for the process of metamorphosis. Therefore, there is possibility as an alternative nutritional supplement for cell culture to reduce the use of FBS. The objective of this study was to evaluate the macrocomponent of hemolymph and the possibility as medium supplement for Spodoptera fugiperda (Sf9) cell culture. Proximate analyses showed that hemolymph contains 89.76% of water, 2.52 mg/mL carbohydrate, 2.35% fat and 55.61 mg/mL protein. Further protein analysis, it consists of 15 fractions containing molecular weight of 22 - 152 kDa. The use of hemolymph as FBS substitution in Sf9 cell culture with various concentrations was unable to maintain and support cell growth. Further research still needed by prior adaptation of the tissue culture to minimal nutrition media before introduction of the hemolymph as supplement.

  20. Preparation and characterization of regenerated Bombyx mori silk fibroin fiber with high strength

    Directory of Open Access Journals (Sweden)

    2008-12-01

    Full Text Available Regenerated Bombyx mori silk fibers were spun from hexafluoro-iso- propanol solution of silk fibroin sponge in methanol used as a coagulant solvent and then elongated in water. The stress-strain curves of the regenerated fibers changed dramatically depending on the draw ratio and the structure was studied by 13C CP/MAS NMR and X-ray diffraction methods. The patterns of 13C CP/MAS NMR spectra of two regenerated fibers with different draw ratios (1× and 3× and native silk fiber are all β-sheet structure although the fraction of random coil/distorted β-turn decreases in the order of 1×, 3× and native fiber gradually. On the other hand, azimuthal scans of their X-ray fiber patterns changed remarkably with increasing the draw ratio. This indicates that long-range orientation of the fibroin chain changes remarkably during the drawing process, but the short-range local structure does not change significantly. Regenerated silk fiber with a draw ratio of 3× is a fiber with high strength which is comparable with that of natural silk fiber. The regenerated fiber is also more degradable than natural silk fiber in enzyme solution in vitro.

  1. Induced Hyperproteinemia and Its Effects on the Remodeling of Fat Bodies in Silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Xue-Dong Chen

    2018-03-01

    Full Text Available Hyperproteinemia, which is characterized by an abnormally elevated plasma protein concentration (PPC, is a high-mortality, metabolic complication associated with severe liver and kidney disease. It is difficult to clinically distinguish the difference between the impacts of primary diseases and hyperproteinemia on tissues and organs, and there are no available animal models of hyperproteinemia. Here, we constructed an animal model of hyperproteinemia with a controllable PPC and no primary disease effects in the silkworm Bombyx mori that has attracted interest owing to its potential use in the pathological analysis of model animals. Silkworm have an open circulatory system in which each organ is directly immersed in hemolymph. The fat body (FB of a silkworm, as a major organ for nutrient storage and energy metabolism, can effectively reflect hyperproteinemia-induced metabolic abnormalities in damaged visceral tissues. A pathogenesis study showed that hyperproteinemia attenuated cell autophagy and apoptosis by attenuating an endocrine hormone, thereby preventing FB remodeling during metamorphosis. Meanwhile, hyperproteinemia increased oxidative stress in the FB and resulted in a dysfunction of amino acid conversion. Supplementation with exogenous 20-hydroxyecdysone effectively mitigated the hyperproteinemia-mediated inhibition of FB remodeling.

  2. Characterization and genome comparison of an Indian isolate of bidensovirus infecting the silkworm Bombyx mori.

    Science.gov (United States)

    Gupta, Tania; Ito, Katsuhiko; Kadono-Okuda, Keiko; Murthy, Geetha N; Gowri, E Vijaya; Ponnuvel, Kangayam M

    2018-01-01

    The bipartite genome of an Indian isolate of Bombyx mori bidensovirus (BmBDV), one of the causative agents of the fatal silkworm disease 'Flacherie', was cloned and completely sequenced. Nucleotide sequence analysis of this Indian isolate of BmBDV revealed two viral DNA segments, VD1 and VD2 as well as a DNA polymerase motif which supports its taxonomical status as the type species of a new family of Bidnaviridae. The Indian isolate of BmBDV was found to have a total of six putative ORFs four of which were located on the VD1 with the other two being on the VD2 DNA segment. The VD1 DNA segment was found to code for three non-structural proteins including a viral DNA polymerase as well as one structural protein, while the VD2 DNA segment was found to code for one structural and one non-structural protein, similar to that of the Japanese and Zhenjiang isolates of BmBDV. A BmBDV ORF expression study was done through real time qPCR wherein the VD2 ORF 1 and 2 showed the maximum transcript levels. This is the first report of the genome characterization of an Indian isolate of BmBDV, infecting silkworm B. mori.

  3. Genetic analysis of γ-ray induced W-translocation strain on Bombyx nori

    International Nuclear Information System (INIS)

    Onuma, Akio; Murakami, Akio

    1976-01-01

    In the process of analyzing a γ-ray induced mutant of Bombyx nori oo cyte, new type translocation strains of W chromosomes and No.5 chromosomes were detected. The constitution of their translocated chromosomes was assumed to be Z/(W-V) + sup(pe)-V + sup(oc)/v. Owing to such chromosome constitution, it was considered that non-disjunction was induced at meiosis, and Z/(W-V) + sup(pe)/V, Z/(W-V) + sup(pe), V/V were produced besides Z/(W-V) + sup(pe)-V + sup(oc)/V in the female chromosomes (gene) of the next progeny, while V/V and Z/Z, V + sup(oc)/V were produced besides Z/Z, V/V in male. Death of some male eggs in this translocation strain was also observed. No dissociated individual of translocated chromosomes was segregated in the next progeny of the female moth with Z/(W-V) + sup(pe), V/V chromosome constitution and the marker stock male moth, while a few dissociated individuals appeared in the next progeny of Z/(W-V) + sup(pe)-V + sup(oc)/V female moth group. This fact seemed to be resulted from the complicated translocated chromosome constitution of the translocation strain. (Kobatake, H.)

  4. Cloning, expression and characterization of alcohol dehydrogenases in the silkworm Bombyx mori

    Directory of Open Access Journals (Sweden)

    Nan Wang

    2011-01-01

    Full Text Available Alcohol dehydrogenases (ADH are a class of enzymes that catalyze the reversible oxidation of alcohols to corresponding aldehydes or ketones, by using either nicotinamide adenine dinucleotide (NAD or nicotinamide adenine dinucleotide phosphate (NADP, as coenzymes. In this study, a short-chain ADH gene was identified in Bombyx mori by 5'-RACE PCR. This is the first time the coding region of BmADH has been cloned, expressed, purified and then characterized. The cDNA fragment encoding the BmADH protein was amplified from a pool of silkworm cDNAs by PCR, and then cloned into E. coli expression vector pET-30a(+. The recombinant His-tagged BmADH protein was expressed in E. coli BL21 (DE3, and then purified by metal chelating affinity chromatography. The soluble recombinant BmADH, produced at low-growth temperature, was instrumental in catalyzing the ethanol-dependent reduction of NAD+, thereby indicating ethanol as one of the substrates of BmADH.

  5. Molecular and neural mechanisms of sex pheromone reception and processing in the silkmoth Bombyx mori

    Directory of Open Access Journals (Sweden)

    Takeshi eSakurai

    2014-03-01

    Full Text Available Male moths locate their mates using species-specific sex pheromones emitted by conspecific females. One striking feature of sex pheromone recognition in males is the high degree of specificity and sensitivity at all levels, from the primary sensory processes to behavior. The silkmoth Bombyx mori is an excellent model insect in which to decipher the underlying mechanisms of sex pheromone recognition due to its simple sex pheromone communication system, where a single pheromone component, bombykol, elicits the full sexual behavior of male moths. Various technical advancements that cover all levels of analysis from molecular to behavioral also allow the systematic analysis of pheromone recognition mechanisms. Sex pheromone signals are detected by pheromone receptors expressed in olfactory receptor neurons in the pheromone-sensitive sensilla trichodea on male antennae. The signals are transmitted to the first olfactory processing center, the antennal lobe (AL, and then are processed further in the higher centers (mushroom body and lateral protocerebrum to elicit orientation behavior towards females. In recent years, significant progress has been made elucidating the molecular mechanisms underlying the detection of sex pheromones. In addition, extensive studies of the AL and higher centers have provided insights into the neural basis of pheromone processing in the silkmoth brain. This review describes these latest advances, and discusses what these advances have revealed about the mechanisms underlying the specific and sensitive recognition of sex pheromones in the silkmoth.

  6. Impact of microsporidian infection on growth and development of silkworm Bombyx mori L. (Lepidoptera: Bombycidae

    Directory of Open Access Journals (Sweden)

    Sunil Kumar Gupta

    2016-09-01

    Full Text Available Several species and strains of microsporidia have been isolated from infected silkworms among which pebrine caused by Nosema bombycis Nageli is the most important. Infection from this disease causes severe economic loss in sericulture. Reduction of larval and pupal development and reduced weights in silkworms due to infection has been reported. In the present study, five microsporidian (Nosema isolates from mulberry silkworm, Bombyx mori L. collected from different locations in West Bengal, India were sampled to study the impact of their infection on the growth and development of B. mori. The study revealed significant differences among the isolates in their ability to cause a reduction in the larval and pupal development of silkworm. Healthy larvae showed better body and tissue weights which were significantly higher than in infected lots. Among the isolates, M5 registered the maximum reduction in relative growth rate, larval silk gland tissue somatic index, larval male and female gonad tissue somatic index (GTSI and pupal female GTSI compared to the healthy control. Male and female pupa treated with M5 spores died before emergence, suggesting that the M5 isolate was the most virulent.

  7. FLP Recombinase-Mediated Site-Specific Recombination in Silkworm, Bombyx mori

    Science.gov (United States)

    Long, Ding-Pei; Zhao, Ai-Chun; Chen, Xue-Jiao; Zhang, Yang; Lu, Wei-Jian; Guo, Qing; Handler, Alfred M.; Xiang, Zhong-Huai

    2012-01-01

    A comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they have not yet been established for use in the manipulation of the silkworm Bombyx mori genome. In this study, we achieved site-specific excision of a target gene at predefined chromosomal sites in the silkworm using a FLP/FRT site-specific recombination system. We first constructed two stable transgenic target silkworm strains that both contain a single copy of the transgene construct comprising a target gene expression cassette flanked by FRT sites. Using pre-blastoderm microinjection of a FLP recombinase helper expression vector, 32 G3 site-specific recombinant transgenic individuals were isolated from five of 143 broods. The average frequency of FLP recombinase-mediated site-specific excision in the two target strains genome was approximately 3.5%. This study shows that it is feasible to achieve site-specific recombination in silkworms using the FLP/FRT system. We conclude that the FLP/FRT system is a useful tool for genome manipulation in the silkworm. Furthermore, this is the first reported use of the FLP/FRT system for the genetic manipulation of a lepidopteran genome and thus provides a useful reference for the establishment of genome manipulation technologies in other lepidopteran species. PMID:22768245

  8. Biological and molecular characterization of silkworm strains from the Brazilian germplasm bank of Bombyx mori.

    Science.gov (United States)

    Pereira, N C; Munhoz, R E F; Bignotto, T S; Bespalhuk, R; Garay, L B; Saez, C R N; Fassina, V A; Nembri, A; Fernandez, M A

    2013-06-28

    Brazil has only one public genetic pool of Bombyx mori strains, which was established in 2005 at Universidade Estadual de Maringá, Maringá, Paraná State. This genetic bank has been maintained, and the strains have been characterized using genetic and morphological tools. The quantitative and qualitative traits, directly or indirectly related to productivity, were evaluated in 14 silkworm strains. In addition to biological and productivity analyses, DNA markers related to susceptibility to the B. mori nucleopolyhedrovirus (BmNPV) were analyzed. BmNPV is a major cause of production loss and is a serious problem for Paraná sericulture. The silkworm strains from diverse geographic origins were found to have different characteristics, including body weight, larval stage duration, cocoon weight, and other biological traits. In terms of productivity, the raw silk percentages were almost uniform, with an overall average of 16.28%. Overall, the Chinese strain C37 gave the best performance in many of the quantitative traits, and it surpassed the other strains in productivity traits. Therefore, it can be used as one of the strains that compose the elite germplasm for silkworm breeding programs. Additionally, genetic molecular markers were efficient in discriminating between B. mori strains that had been identified based on their geographical origin. We found that all Japanese strains produced a 400-bp molecular marker that has been associated with susceptibility to BmNPV.

  9. The progress and future of enhancing antiviral capacity by transgenic technology in the silkworm Bombyx mori.

    Science.gov (United States)

    Jiang, Liang; Xia, Qingyou

    2014-05-01

    Bombyx mori is a common lepidopteran model and an important economic insect for silk production. B. mori nucleopolyhedrovirus (BmNPV) is a typical pathogenic baculovirus that causes serious economic losses in sericulture. B. mori and BmNPV are a model of insect host and pathogen interaction including invasion of the host by the pathogen, host response, and enhancement of host resistance. The antiviral capacity of silkworms can be improved by transgenic technology such as overexpression of an endogenous or exogenous antiviral gene, RNA interference of the BmNPV gene, or regulation of the immune pathway to inhibit BmNPV at different stages of infection. Antiviral capacity could be further increased by combining different methods. We discuss the future of an antiviral strategy in silkworm, including possible improvement of anti-BmNPV, the feasibility of constructing transgenic silkworms with resistance to multiple viruses, and the safety of transgenic silkworms. The silkworm model could provide a reference for disease control in other organisms. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  10. Characterization of Some Breeds and Hybrids of Bombyx mori L. Silkworms through Biological Index of Spawning

    Directory of Open Access Journals (Sweden)

    Hotatiu Dezmirean

    2015-05-01

    Full Text Available The main aim of this study is to determine the parameters of biological indexes of Bombyx mori L. spawning, in order to obtain adequate larvae for experimental breeding using MSF (Familial Sericulture Modulus. The biological material used for experiments contains the pure breeds AB (Alb Băneasa, B1 (Băneasa 1, AC (Alb de Cislău, AC29 (Alb chinezesc, line 29, AC29/T (Alb Chinezesc, AC/T (Alb de Cislău, RG90, IBV, S8, AJ5/F, C127, AJ17, C108/T, AO/T and the hybrids B1XAC, ACXB1,  ABXAC29, AC29XAB, ACXAB, ABXAC, ABXB1, B1XAB. For each pure breed and hybrid were determinate the number of eggs/spawning, the number of hatching eggs, the hatching percent (% and the number of unhatching eggs. The analyse of these indexes was done in three consecutive years, and the averages and spread parameters, as well as variability coefficients (V% were calculated for each breed and hybrid. The obtained values of biological indexes of spawning were compared with those reported in autochthon and foreigner literature. The main conclusions of our study are that the autochthon breeds and hybrids values are according to the values published by other researchers and correspond for the experimental breeding.

  11. Genetic Characterization of Bombyx mori (Lepidoptera: Bombycidae) Breeding and Hybrid Lines With Different Geographic Origins

    Science.gov (United States)

    Furdui, Emilia M.; Mărghitaş, Liviu A.; Dezmirean, Daniel S.; Paşca, Ioan; Pop, Iulia F.; Erler, Silvio; Schlüns, Ellen A.

    2014-01-01

    Abstract The domesticated silkworm Bombyx mori L. comprises a large number of geographical breeds and hybrid lines. Knowing the genetic structure of those may provide information to improve the conservation of commercial lines by estimating inbreeding over generations and the consequences of excessive use of those lineages. Here, we analyzed the genetic diversity of seven breeds and eight hybrid lines from Eastern Europe and Asia using highly polymorphic microsatellites markers to determine its genetical impact on their use in global breeding programs. No consistent pattern of deviation from Hardy–Weinberg equilibrium was found for most breed and hybrids; and the absence of a linkage disequilibrium also suggests that the strains are in equilibrium. A principal coordinate analysis revealed a clear separation of two silkworm breeds from the rest: one (IBV) originated from India and the other one (RG 90 ) from Romania/Japan. The tendency of the other breeds from different geographic origins to cluster together in a general mix might be due to similar selection pressures (climate and anthropogenic factors) in different geographic locations. Phylogenetic analyses grouped the different silkworm breeds but not the hybrids according to their geographic origin and confirmed the pattern found in the principal coordinate analysis. PMID:25502023

  12. Metabolomics differences between silkworms (Bombyx mori) reared on fresh mulberry (Morus) leaves or artificial diets.

    Science.gov (United States)

    Dong, Hui-Ling; Zhang, Sheng-Xiang; Tao, Hui; Chen, Zhuo-Hua; Li, Xue; Qiu, Jian-Feng; Cui, Wen-Zhao; Sima, Yang-Hu; Cui, Wei-Zheng; Xu, Shi-Qing

    2017-09-08

    Silkworms (Bombyx mori) reared on artificial diets have great potential applications in sericulture. However, the mechanisms underlying the enhancement of metabolic utilization by altering silkworm nutrition are unclear. The aim of this study was to investigate the mechanisms responsible for the poor development and low silk protein synthesis efficiency of silkworms fed artificial diets. After multi-generational selection of the ingestive behavior of silkworms to artificial diets, we obtained two strains, one of which developed well and another in which almost all its larvae starved to death on the artificial diets. Subsequently, we analyzed the metabolomics of larval hemolymph by gas chromatography/liquid chromatography-mass spectrometry, and the results showed that vitamins were in critically short supply, whereas the nitrogen metabolic end product of urea and uric acid were enriched substantially, in the hemolymph of the silkworms reared on the artificial diets. Meanwhile, amino acid metabolic disorders, as well as downregulation of carbohydrate metabolism, energy metabolism, and lipid metabolism, co-occurred. Furthermore, 10 male-dominant metabolites and 27 diet-related metabolites that differed between male and female silkworms were identified. These findings provide important insights into the regulation of silkworm metabolism and silk protein synthesis when silkworms adapt to an artificial diet.

  13. Knockdown of Broad-Complex Gene Expression of Bombyx mori by Oligopyrrole Carboxamides Enhances Silk Production.

    Science.gov (United States)

    Ali, Asfa; Bovilla, Venugopal Reddy; Mysarla, Danti Kumari; Siripurapu, Prasanthi; Pathak, Rashmi U; Basu, Bhakti; Mamillapalli, Anitha; Bhattacharya, Santanu

    2017-04-11

    Bombyx mori (B. mori) is important due to its major role in the silk production. Though DNA binding ligands often influence gene expression, no attempt has been made to exploit their use in sericulture. The telomeric heterochromatin of B. mori is enriched with 5'-TTAGG-3' sequences. These sequences were also found to be present in several genes in the euchromatic regions. We examined three synthetic oligopyrrole carboxamides that target 5'-TTAGG-3' sequences in controlling the gene expression in B. mori. The ligands did not show any defect or feeding difference in the larval stage, crucial for silk production. The ligands caused silencing of various isoforms of the broad-complex transcription factor and cuticle proteins which resulted in late pupal developmental defects. Furthermore, treatment with such drugs resulted in statistically enhanced cocoon weight, shell weight, and silk yield. This study shows for the first time use of oligopyrrole carboxamide drugs in controlling gene expression in B. mori and their long term use in enhancing silk production.

  14. Identification of a new Bombyx mori nucleopolyhedrovirus and analysis of its bro gene family.

    Science.gov (United States)

    Zhou, Jing-Bo; Li, Xing-Qi; De-Eknamkul, Wanchai; Suraporn, Siripuk; Xu, Jia-Ping

    2012-06-01

    The highly pathogenic Bombyx mori nucleopolyhedrovirus (BmNPV) has caused severe damages to sericulture in many countries, and the relationship between the pathogenicity of various BmNPV strains and their geographical evolution has been the topic of our interest. In this study, we isolated a new BmNPV strain from Thailand (BmNPV-Thai), based on the sequences of its conservative genes p10, p35, polh, egt and vp39. The BmNPV-Thai appears to have baculovirus repeated ORF (bro) genes different from four other well-known BmNPV strains of China (GD, CQ1), Japan (T3), and France (SC7); It only has bro-a, bro-c, and bro-d, but not bro-b and bro-e genes. These bro genes are localized only in the two subgroups highly homologous to their counterparts and their encoded BRO proteins differ mainly at their N-terminal amino acid residues. Phylogenetic analysis indicates that the evolution of the bro genes of the five BmNPV strains is not obviously associated with their geographic locations.

  15. Differentially expressed microRNAs in diapausing versus HCl-treated Bombyx embryos

    Science.gov (United States)

    Qin, Mingyue; Lin, Bimin; Chen, Fangyan; Yan, Huichao; Li, Wenchu

    2017-01-01

    Differentially expressed microRNAs were detected to explore the molecular mechanisms of diapause termination. The total small RNA of diapause-destined silkworm eggs and HCl-treated eggs was extracted and then sequenced using HiSeq high-throughput method. 44 novel miRNAs were discovered. Compared to those in the diapause-destined eggs, 61 miRNAs showed significant changes in the acid-treated eggs, with 23 being up-regulated and 38 being down-regulated. The potential target genes of differentially expressed miRNAs were predicted by miRanda. Gene Ontology and KEGG pathway enrichment analysis of these potential target genes revealed that they were mainly located within cells and organelles, involved in cellular and metabolic processes, and participated in protein production, processing and transportation. Two differentially expressed genes, Bombyx mori SDH and Bmo-miR-2761-3p, were further analyzed with qRT-PCR. BmSDH was significantly up-regulated in the HCl-treated eggs, while Bmo-miR-2761-3p was down-regulated. These results suggested that these two genes were well coordinated in silkworm eggs. Dual luciferase reporter assay demonstrated that Bmo-miR-2761-3p inhibited the expression of BmSDH. PMID:28700597

  16. Differentially expressed genes in the fat body of Bombyx mori in response to phoxim insecticide.

    Science.gov (United States)

    Gu, Z Y; Li, F C; Wang, B B; Xu, K Z; Ni, M; Zhang, H; Shen, W D; Li, B

    2015-01-01

    The silkworm, Bombyx mori, is an economically important insect. However, poisoning of silkworms by organophosphate pesticides causes tremendous loss to the sericulture. The fat body is the major tissue involved in detoxification and produces antimicrobial peptides and regulates hormones. In this study, a microarray system comprising 22,987 oligonucluotide 70-mer probes was employed to examine differentially expressed genes in the fat body of B. mori exposed to phoxim insecticide. The results showed that a total of 774 genes were differentially expressed upon phoxim exposure, including 500 up-regulated genes and 274 down-regulated genes. The expression levels of eight detoxification-related genes were up-regulated upon phoxim exposure, including six cytochrome P450s and two glutathione-S-transferases. It was firstly found that eight antimicrobial peptide genes were down-regulated, which might provide important references for studying the larvae of B. mori become more susceptible to microbial infections after phoxim treatment. In addition, we firstly detected the expression level of metamorphosis-related genes after phoxim exposure, which may lead to impacted reproduction. Our results may facilitate the overall understanding of the molecular mechanism of multiple pathways following exposure to phoxim insecticide in the fat body of B. mori. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. A novel protease inhibitor in Bombyx mori is involved in defense against Beauveria bassiana.

    Science.gov (United States)

    Li, Youshan; Zhao, Ping; Liu, Shiping; Dong, Zhaoming; Chen, Jianping; Xiang, Zhonghuai; Xia, Qingyou

    2012-10-01

    Entomopathogenic fungi, such as Beauveria bassiana, penetrate the insect cuticle using a plethora of hydrolytic enzymes including cuticle-degrading proteases and chitinases, which are important virulence factors. The insect integument and hemolymph contains a relatively high concentration of protease inhibitors, which are closely involved with defense against pathogenic microorganisms. To elucidate the molecular mechanism underlying resistance against entomopathogenic fungi and to identify a new molecular target for improving fungal resistance in the silkworm, Bombyx mori, we cloned and expressed a novel silkworm TIL-type protease inhibitor BmSPI38, which was very stable over a wide range of temperatures and pH values. An activity assay suggested that BmSPI38 potently inactivated the insecticidal cuticle-degrading enzyme (CDEP-1) produced by B. bassiana and subtilisin A produced by Bacillus licheniformis. The melanization of silkworm induced by CDEP-1 protease could also be blocked by BmSPI38. These results provided new insights into the molecular mechanisms whereby insect protease inhibitors provide resistance against entomopathogenic fungi, suggesting the possibility of using fungal biopesticides in sericulture. Copyright © 2012 Elsevier Ltd. All rights reserved.

  18. Pertumbuhan dan Produktivitas Ulat Sutera Bombyx Mori L. (Lepidoptera : Bombicidae) yang Diberi Vitamin B1 Pada Daun Murbei Morus sp.

    OpenAIRE

    Lubis, Umi Kalsum

    2014-01-01

    The effect of mulberry leaves that contain B1 on the growth and productivity of silkworm (Bombyx mori L.; Lepidoptera; Bombicidae) was conducted in the Laboratory of Genetics, Department of Biology, Faculty of Mathematics and Natural Sciences, University of Sumatera Utara Medan. This study used an experimental method to completely randomized design (RAL). The treatments were vitamine B1 0.0mg/100ml; 0.1mg/100ml, 0.2mg/100ml, 0.3mg/100ml; 0.4 mg/100ml with 3 replications each replication consi...

  19. Establishment of Tools for Neurogenetic Analysis of Sexual Behavior in the Silkmoth, Bombyx mori

    Science.gov (United States)

    Kiya, Taketoshi; Morishita, Koudai; Uchino, Keiro; Iwami, Masafumi; Sezutsu, Hideki

    2014-01-01

    Background Silkmoth, Bombyx mori, is an ideal model insect for investigating the neural mechanisms underlying sex pheromone-induced innate behavior. Although transgenic techniques and the GAL4/UAS system are well established in the silkmoth, genetic tools useful for investigating brain function at the neural circuit level have been lacking. Results In the present study, we established silkmoth strains in which we could visualize neural projections (UAS-mCD8GFP) and cell nucleus positions (UAS-GFP.nls), and manipulate neural excitability by thermal stimulation (UAS-dTrpA1). In these strains, neural projections and nucleus position were reliably labeled with green fluorescent protein in a GAL4-dependent manner. Further, the behavior of silkworm larvae and adults could be controlled by GAL4-dependent misexpression of dTrpA1. Ubiquitous dTrpA1 misexpression led both silkmoth larvae and adults to exhibit seizure-like phenotypes in a heat stimulation-dependent manner. Furthermore, dTrpA1 misexpression in the sex pheromone receptor neurons of male silkmoths allowed us to control male sexual behavior by changing the temperature. Thermally stimulated male silkmoths exhibited full sexual behavior, including wing-flapping, orientation, and attempted copulation, and precisely approached a thermal source in a manner similar to male silkmoths stimulated with the sex pheromone. Conclusion These findings indicate that a thermogenetic approach using dTrpA1 is feasible in Lepidopteran insects and thermogenetic analysis of innate behavior is applicable in the silkmoth. These tools are essential for elucidating the relationships between neural circuits and function using neurogenetic methods. PMID:25396742

  20. Effects of BmCPV Infection on Silkworm Bombyx mori Intestinal Bacteria.

    Directory of Open Access Journals (Sweden)

    Zhenli Sun

    Full Text Available The gut microbiota has a crucial role in the growth, development and environmental adaptation in the host insect. The objective of our work was to investigate the microbiota of the healthy silkworm Bombyx mori gut and changes after the infection of B. mori cypovirus (BmCPV. Intestinal contents of the infected and healthy larvae of B. mori of fifth instar were collected at 24, 72 and 144 h post infection with BmCPV. The gut bacteria were analyzed by pyrosequencing of the 16S rRNA gene. 147(135 and 113(103 genera were found in the gut content of the healthy control female (male larvae and BmCPV-infected female (male larvae, respectively. In general, the microbial communities in the gut content of healthy larvae were dominated by Enterococcus, Delftia, Pelomonas, Ralstonia and Staphylococcus, however the abundance change of each genus was depended on the developmental stage and gender. Microbial diversity reached minimum at 144 h of fifth instar larvae. The abundance of Enterococcus in the females was substantially lower and the abundance of Delftia, Aurantimonas and Staphylococcus was substantially higher compared to the males. Bacterial diversity in the intestinal contents decreased after post infection with BmCPV, whereas the abundance of both Enterococcus and Staphylococcus which belongs to Gram-positive were increased. Therefore, our findings suggested that observed changes in relative abundance was related to the immune response of silkworm to BmCPV infection. Relevance analysis of plenty of the predominant genera showed the abundance of the Enterococcus genus was in negative correlation with the abundance of the most predominant genera. These results provided insight into the relationship between the gut microbiota and development of the BmCPV-infected silkworm.

  1. Establishment of tools for neurogenetic analysis of sexual behavior in the silkmoth, Bombyx mori.

    Science.gov (United States)

    Kiya, Taketoshi; Morishita, Koudai; Uchino, Keiro; Iwami, Masafumi; Sezutsu, Hideki

    2014-01-01

    Silkmoth, Bombyx mori, is an ideal model insect for investigating the neural mechanisms underlying sex pheromone-induced innate behavior. Although transgenic techniques and the GAL4/UAS system are well established in the silkmoth, genetic tools useful for investigating brain function at the neural circuit level have been lacking. In the present study, we established silkmoth strains in which we could visualize neural projections (UAS-mCD8GFP) and cell nucleus positions (UAS-GFP.nls), and manipulate neural excitability by thermal stimulation (UAS-dTrpA1). In these strains, neural projections and nucleus position were reliably labeled with green fluorescent protein in a GAL4-dependent manner. Further, the behavior of silkworm larvae and adults could be controlled by GAL4-dependent misexpression of dTrpA1. Ubiquitous dTrpA1 misexpression led both silkmoth larvae and adults to exhibit seizure-like phenotypes in a heat stimulation-dependent manner. Furthermore, dTrpA1 misexpression in the sex pheromone receptor neurons of male silkmoths allowed us to control male sexual behavior by changing the temperature. Thermally stimulated male silkmoths exhibited full sexual behavior, including wing-flapping, orientation, and attempted copulation, and precisely approached a thermal source in a manner similar to male silkmoths stimulated with the sex pheromone. These findings indicate that a thermogenetic approach using dTrpA1 is feasible in Lepidopteran insects and thermogenetic analysis of innate behavior is applicable in the silkmoth. These tools are essential for elucidating the relationships between neural circuits and function using neurogenetic methods.

  2. Establishment of tools for neurogenetic analysis of sexual behavior in the silkmoth, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Taketoshi Kiya

    Full Text Available Silkmoth, Bombyx mori, is an ideal model insect for investigating the neural mechanisms underlying sex pheromone-induced innate behavior. Although transgenic techniques and the GAL4/UAS system are well established in the silkmoth, genetic tools useful for investigating brain function at the neural circuit level have been lacking.In the present study, we established silkmoth strains in which we could visualize neural projections (UAS-mCD8GFP and cell nucleus positions (UAS-GFP.nls, and manipulate neural excitability by thermal stimulation (UAS-dTrpA1. In these strains, neural projections and nucleus position were reliably labeled with green fluorescent protein in a GAL4-dependent manner. Further, the behavior of silkworm larvae and adults could be controlled by GAL4-dependent misexpression of dTrpA1. Ubiquitous dTrpA1 misexpression led both silkmoth larvae and adults to exhibit seizure-like phenotypes in a heat stimulation-dependent manner. Furthermore, dTrpA1 misexpression in the sex pheromone receptor neurons of male silkmoths allowed us to control male sexual behavior by changing the temperature. Thermally stimulated male silkmoths exhibited full sexual behavior, including wing-flapping, orientation, and attempted copulation, and precisely approached a thermal source in a manner similar to male silkmoths stimulated with the sex pheromone.These findings indicate that a thermogenetic approach using dTrpA1 is feasible in Lepidopteran insects and thermogenetic analysis of innate behavior is applicable in the silkmoth. These tools are essential for elucidating the relationships between neural circuits and function using neurogenetic methods.

  3. A DNA Binding Protein Is Required for Viral Replication and Transcription in Bombyx mori Nucleopolyhedrovirus.

    Science.gov (United States)

    Zhao, Cui; Zhang, Chen; Chen, Bin; Shi, Yanghui; Quan, Yanping; Nie, Zuoming; Zhang, Yaozhou; Yu, Wei

    2016-01-01

    A DNA-binding protein (DBP) [GenBank accession number: M63416] of Bombyx mori nuclear polyhedrosis virus (BmNPV) has been reported to be a regulatory factor in BmNPV, but its detailed functions remain unknown. In order to study the regulatory mechanism of DBP on viral proliferation, genome replication, and gene transcription, a BmNPV dbp gene knockout virus dbp-ko-Bacmid was generated by the means of Red recombination system. In addition, dbp-repaired virus dbp-re-Bacmid was constructed by the means of the Bac to Bac system. Then, the Bacmids were transfected into BmN cells. The results of this viral titer experiment revealed that the TCID50 of the dbp-ko-Bacmid was 0; however, the dbp-re-Bacmid was similar to the wtBacmid (p>0.05), indicating that the dbp-deficient would lead to failure in the assembly of virus particles. In the next step, Real-Time PCR was used to analyze the transcriptional phases of dbp gene in BmN cells, which had been infected with BmNPV. The results of the latter experiment revealed that the transcript of dbp gene was first detected at 3 h post-infection. Furthermore, the replication level of virus genome and the transcriptional level of virus early, late, and very late genes in BmN cells, which had been transfected with 3 kinds of Bacmids, were analyzed by Real-Time PCR. The demonstrating that the replication level of genome was lower than that of wtBacmid and dbp-re-Bacmid (pviral replication, but also a viral gene that has a significant impact on transcription and expression during all periods of baculovirus life cycle.

  4. Effect of the sterilization method on the properties of Bombyx mori silk fibroin films

    Energy Technology Data Exchange (ETDEWEB)

    George, Karina A. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland 4059 (Australia); Shadforth, Audra M.A. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Chirila, Traian V., E-mail: traian.chirila@qei.org.au [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Australian Institute of Bioengineering and Nanotechnology, University of Queensland, St Lucia, 4072 (Australia); Faculty of Health Sciences, University of Queensland, Herston, Queensland 4006 (Australia); Faculty of Science and Engineering, Queensland University of Technology, Brisbane, Queensland 4001 (Australia); Laurent, Matthieu J. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Ecole Superieure d' Ingenieurs de Luminy (ESIL), Universite de la Mediterranee Aix-Marseille II, Luminy case 925 13288, Marseille, Cedex 09 (France); Stephenson, Sally-Anne [Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland 4059 (Australia); Faculty of Health, Queensland University of Technology, Brisbane, Queensland 4001 (Australia); Edwards, Grant A. [Australian Institute of Bioengineering and Nanotechnology, University of Queensland, St Lucia, 4072 (Australia); Madden, Peter W. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Faculty of Health Sciences, University of Queensland, Herston, Queensland 4006 (Australia); and others

    2013-03-01

    We have compared the effects of different sterilization techniques on the properties of Bombyx mori silk fibroin thin films with the view to subsequent use for corneal tissue engineering. The transparency, tensile properties, corneal epithelial cell attachment and degradation of the films were used to evaluate the suitability of certain sterilization techniques including gamma-irradiation (in air or nitrogen), steam treatment and immersion in aqueous ethanol. The investigations showed that gamma-irradiation, performed either in air or in a nitrogen atmosphere, did not significantly alter the properties of films. The films sterilized by gamma-irradiation or by immersion in ethanol had a transparency greater than 98% and tensile properties comparable to human cornea and amniotic membrane, the materials of choice in the reconstruction of ocular surface. Although steam-sterilization produced stronger, stiffer films, they were less transparent, and cell attachment was affected by the variable topography of these films. It was concluded that gamma-irradiation should be considered to be the most suitable method for the sterilization of silk fibroin films, however, the treatment with ethanol is also an acceptable method. - Highlights: Black-Right-Pointing-Pointer The effects of four methods of sterilization on the properties of silk fibroin films were investigated. Black-Right-Pointing-Pointer Steam treatment leads to stiffer films but to lower transparency and variable surface topography. Black-Right-Pointing-Pointer Degradation of fibroin is enhanced in the films that were gamma-irradiated. Black-Right-Pointing-Pointer The effects on mechanical properties are explained through changes in both primary and secondary structure of fibroin. Black-Right-Pointing-Pointer Gamma-irradiation and immersion in aqueous ethanol are suggested as preferred methods of sterilization.

  5. Exogenous gene can be expressed by a recombinant Bombyx mori cypovirus.

    Science.gov (United States)

    Guo, Rui; Cao, Guangli; Xue, Renyu; Kumar, Dhiraj; Chen, Fei; Liu, Wei; Jiang, Yue; Lu, Yahong; Zhu, Liyuan; Liang, Zi; Kuang, Sulan; Hu, Xiaolong; Gong, Chengliang

    2018-02-01

    Bombyx mori cypovirus (BmCPV) is one of the major viral pathogen for silkworm, and the genome of BmCPV is composed of 10 dsRNA segments. As construction system of recombinant BmCPV (rBmCPV) is scanty, researchers achieved little progress in studying gene function of BmCPV in recent decades. Here, 10 recombinant plasmids with a full-length cDNA of viral genome segments S1-S10 containing T7 promoter were constructed. After cotransfecting the BmN cells with the mixture of 10 in vitro-transcribed RNAs, pathological changes were observed. Real-time PCR and Western blot showed viral gene vp1 and structural proteins were expressed. It is found the genome of the rBmCPV is composed of 10 dsRNA segments similar to those of wild-type BmCPV. Moreover, viral particles and polyhedron with virions can be generated in the cotransfected cells and the injected silkworm midguts. These findings confirmed the formation of infective rBmCPV. Additionally, we found viable rBmCPV was generated by cotransfecting the mixture of in vitro-transcribed S1-S9 RNAs into the cultured cells, confirming polh was not essential for BmCPV replication. Moreover, an infectious rBmCPV expressing the DsRed protein was constructed based on this system. Further investigation showed S2 and S7 segments are indispensible for viral proliferation. Our findings demonstrated the construction system of rBmCPV can be utilized for exploring viral replication and pathogenesis, and investigated method for constructing rBmCPV will certainly facilitate developing novel biopesticides and expressing exogenous gene in the midgut of silkworm.

  6. Precocious metamorphosis in the juvenile hormone-deficient mutant of the silkworm, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Takaaki Daimon

    Full Text Available Insect molting and metamorphosis are intricately governed by two hormones, ecdysteroids and juvenile hormones (JHs. JHs prevent precocious metamorphosis and allow the larva to undergo multiple rounds of molting until it attains the proper size for metamorphosis. In the silkworm, Bombyx mori, several "moltinism" mutations have been identified that exhibit variations in the number of larval molts; however, none of them have been characterized molecularly. Here we report the identification and characterization of the gene responsible for the dimolting (mod mutant that undergoes precocious metamorphosis with fewer larval-larval molts. We show that the mod mutation results in complete loss of JHs in the larval hemolymph and that the mutant phenotype can be rescued by topical application of a JH analog. We performed positional cloning of mod and found a null mutation in the cytochrome P450 gene CYP15C1 in the mod allele. We also demonstrated that CYP15C1 is specifically expressed in the corpus allatum, an endocrine organ that synthesizes and secretes JHs. Furthermore, a biochemical experiment showed that CYP15C1 epoxidizes farnesoic acid to JH acid in a highly stereospecific manner. Precocious metamorphosis of mod larvae was rescued when the wild-type allele of CYP15C1 was expressed in transgenic mod larvae using the GAL4/UAS system. Our data therefore reveal that CYP15C1 is the gene responsible for the mod mutation and is essential for JH biosynthesis. Remarkably, precocious larval-pupal transition in mod larvae does not occur in the first or second instar, suggesting that authentic epoxidized JHs are not essential in very young larvae of B. mori. Our identification of a JH-deficient mutant in this model insect will lead to a greater understanding of the molecular basis of the hormonal control of development and metamorphosis.

  7. Development and reproduction of Podisus distinctus (Heteroptera: Pentatomidae fed on larva of Bombyx mori (Lepidoptera: Bombycidae

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    M. C. Lacerda

    Full Text Available Biological control has been reducing the use of chemical products against insect pests, specially predatory Pentatomidae. Species of this group can present high variations in their life cycle as a result of their diet. Thus, the objective of this research was to study nymph development and reproduction of Podisus distinctus (Stäl, 1860 (Heteroptera: Pentatomidae fed on Bombyx mori L., 1758 (Lepidoptera: Bombycidae larvae (T1, compared to those fed on Tenebrio molitor L., 1758 (Coleoptera: Tenebrionidae (T2 and Musca domestica L., 1758 (Diptera: Muscidae larvae (T3 at a temperature of 25 ± 0.5ºC, relative humidity of 70 ± 2%, and photophase of 12 h. Predators fed on B. mori showed duration of the nymph phase (18.68 ± 1.02 similar to those fed on T. molitor (18.32 ± 1.49. Pre-oviposition and oviposition periods and number of egg masses, besides eggs and nymphs per female, were higher with B. mori (5.83 ± 2.02; 15.00 ± 7.40; 8.42 ± 1.84; 296.69 ± 154.75; and 228.55 ± 141.04, respectively while longevity of males and females of P. distinctus was 25.76 ± 16.15 and 35.00 ± 16.15 days with T. molitor, and 20.57 ± 13.60 and 23.46 ± 12.35 days with B. mori, respectively.

  8. Nutrigenetic screening strains of the mulberry silkworm, Bombyx mori, for nutritional efficiency.

    Science.gov (United States)

    Chinnaswamy, Ramesha; Lakshmi, Hothur; Kumari, Savarapu S; Anuradha, Chebba M; Kumar, Chitta S

    2012-01-01

    The activity of sericulture is declining due the reduction of mulberry production area in sericulture practicing countries lead to adverse effects on silkworm rearing and cocoon production. Screening for nutrigenetic traits in silkworm, Bombyx mori L. (Lepidoptera: Bombycidae) is an essential prerequisite for better understanding and development of nutritionally efficient breeds/hybrids, which show less food consumption with higher efficiency conversion. The aim of this study was to identify nutritionally efficient polyvoltine silkworm strains using the germplasm breeds RMW(2), RMW(3), RMW(4), RMG(3), RMG(1), RMG(4), RMG(5), RMG(6) and APM(1) as the control. The 1(st) day of 5(th) stage silkworm larvae of polyvoltine strains were subjected to standard gravimetric analysis until spinning for three consecutive generations covering three different seasons on 19 nutrigenetic traits. Highly significant (p ≤ 0.001) differences were found among all nutrigenetic traits of polyvoltine silkworm strains in the experimental groups. The nutritionally efficient polvoltine silkworm strains were resulted by utilizing nutrition consumption index and efficiency of conversion of ingesta/cocoon traits as the index. Higher nutritional efficiency conversions were found in the polyvoltine silkworm strains on efficiency of conversion of ingesta to cocoon and shell than control. Comparatively smaller consumption index, respiration, metabolic rate with superior relative growth rate, and quantum of food ingesta and digesta requisite per gram of cocoon and shell were shown; the lowest amount was in new polyvoltine strains compared to the control. Furthermore, based on the overall nutrigenetic traits utilized as index or 'biomarkers', three polyvoltine silkworm strains (RMG(4), RMW(2), and RMW(3)) were identified as having the potential for nutrition efficiency conversion. The data from the present study advances our knowledge for the development of nutritionally efficient silkworm breeds

  9. Stimulation of JNK Phosphorylation by the PTTH in Prothoracic Glands of the Silkworm, Bombyx mori

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    Shi-Hong Gu

    2018-02-01

    Full Text Available In this study, phosphorylation of c-Jun N-terminal kinase (JNK by the prothoracicotropic hormone (PTTH was investigated in prothoracic glands (PGs of the silkworm, Bombyx mori. Results showed that JNK phosphorylation was stimulated by the PTTH in time- and dose-dependent manners. In vitro activation of JNK phosphorylation in PGs by the PTTH was also confirmed in an in vivo experiment, in which a PTTH injection greatly increased JNK phosphorylation in PGs of day-6 last instar larvae. JNK phosphorylation caused by PTTH stimulation was greatly inhibited by U73122, a potent and specific inhibitor of phospholipase C (PLC and an increase in JNK phosphorylation was also detected when PGs were treated with agents (either A23187 or thapsigargin that directly elevated the intracellular Ca2+ concentration, thereby indicating involvement of PLC and Ca2+. Pretreatment with an inhibitor (U0126 of mitogen-activated protein kinase (MAPK/extracellular signal-regulated kinase (ERK kinase (MEK and an inhibitor (LY294002 of phosphoinositide 3-kinase (PI3K failed to significantly inhibit PTTH-stimulated JNK phosphorylation, indicating that ERK and PI3K were not related to JNK. We further investigated the effect of modulation of the redox state on JNK phosphorylation. In the presence of either an antioxidant (N-acetylcysteine, NAC or diphenylene iodonium (DPI, PTTH-stimulated JNK phosphorylation was blocked. The JNK kinase inhibitor, SP600125, markedly inhibited PTTH-stimulated JNK phosphorylation and ecdysteroid synthesis. The kinase assay of JNK in PGs confirmed its stimulation by PTTH and inhibition by SP600125. Moreover, PTTH treatment did not affect JNK or Jun mRNA expressions. Based on these findings, we concluded that PTTH stimulates JNK phosphorylation in Ca2+- and PLC-dependent manners and that the redox-regulated JNK signaling pathway is involved in PTTH-stimulated ecdysteroid synthesis in B. mori PGs.

  10. Identification of specific sites in the third intracellular loop and carboxyl terminus of the Bombyx mori PBAN receptor crucial for ligand-induced internalization

    Science.gov (United States)

    Sex pheromone production in most moths is mediated by the pheromone biosynthesis activating neuropeptide receptor (PBANR). Similar to other rhodopsin-like G protein-coupled receptors, the silkmoth Bombyx mori PBANR (BmPBANR) undergoes agonist-induced internalization. Despite interest in developing...

  11. The molecular mobility of water in natural polymers : Silk Bombyx mori with a low water content as studied by H-1 DQF NMR

    NARCIS (Netherlands)

    Rodin, VV; Knight, DP

    2004-01-01

    The molecular mobility of water in fibres of natural silk (Bombyx mori) was studied by the double-quantum-filtered (DQF) and single-pulse H-1 NMR techniques. The results obtained showed a slow motion of water molecules and their strong interaction with silk macromolecules. At different model

  12. Genome Sequence of Enterococcus mundtii EM01, Isolated from Bombyx mori Midgut and Responsible for Flacherie Disease in Silkworms Reared on an Artificial Diet

    DEFF Research Database (Denmark)

    de Diego-Diaz, Beatriz; Treu, Laura; Campanaro, Stefano

    2018-01-01

    The whole genome sequence of Enterococcus mundtii strain EM01 is reported here. The isolate proved to be the cause of flacherie in Bombyx mori To date, the genomes of 11 other E. mundtii strains have been sequenced. EM01 is the only strain that displayed active pathological effects on its...... associated animal species....

  13. Localization and functional analysis of the insect-specific RabX4 in the brain of Bombyx mori.

    Science.gov (United States)

    Uno, Tomohide; Furutani, Masayuki; Sakamoto, Katsuhiko; Uno, Yuichi; Kanamaru, Kengo; Mizoguchi, Akira; Hiragaki, Susumu; Takeda, Makio

    2017-09-01

    Rab proteins are small monomeric GTPases/GTP-binding proteins, which form the largest branch of the Ras superfamily. The different Rab GTPases are localized to the cytosolic face of specific intracellular membranes, where they function as regulators of distinct steps in membrane trafficking. RabX4 is an insect-specific Rab protein that has no close homolog in vertebrates. There is little information about insect-specific Rab proteins. RabX4 was expressed in Escherichia coli and subsequently purified. Antibodies against Bombyx mori RabX4 were produced in rabbits for western immunoblotting and immunohistochemistry. Western blotting of neural tissues revealed a single band, at approximately 26 kD. RabX4-like immunohistochemical reactivity was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum in the brain. Further immunohistochemical analysis revealed that RabX4 colocalized with Rab6 and bombyxin in the corpus allatum, a neuronal organ that secretes neuropeptides synthesized in the brain into the hemolymph. RabX4 expression in the frontal ganglion, part of the insect stomatogastric nervous system that is found in most insect orders, was restricted to two neurons on the outer region and did not colocalize with allatotropin or Rab6. Furthermore, RNA interference of RabX4 decreased bombyxin expression levels in the brain. These findings suggest that RabX4 is involved in the neurosecretion of a secretory organ in Bombyx mori. © 2017 Wiley Periodicals, Inc.

  14. Regulation of Silk Genes by Hox and Homeodomain Proteins in the Terminal Differentiated Silk Gland of the Silkworm Bombyx mori

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    Shigeharu Takiya

    2016-05-01

    Full Text Available The silk gland of the silkworm Bombyx mori is a long tubular organ that is divided into several subparts along its anteroposterior (AP axis. As a trait of terminal differentiation of the silk gland, several silk protein genes are expressed with unique regional specificities. Most of the Hox and some of the homeobox genes are also expressed in the differentiated silk gland with regional specificities. The expression patterns of Hox genes in the silk gland roughly correspond to those in embryogenesis showing “colinearity”. The central Hox class protein Antennapedia (Antp directly regulates the expression of several middle silk gland–specific silk genes, whereas the Lin-1/Isl-1/Mec3 (LIM-homeodomain transcriptional factor Arrowhead (Awh regulates the expression of posterior silk gland–specific genes for silk fiber proteins. We summarize our results and discuss the usefulness of the silk gland of Bombyx mori for analyzing the function of Hox genes. Further analyses of the regulatory mechanisms underlying the region-specific expression of silk genes will provide novel insights into the molecular bases for target-gene selection and regulation by Hox and homeodomain proteins.

  15. Binding specificity of Bacillus thuringiensis Cry1Aa for purified, native Bombyx mori aminopeptidase N and cadherin-like receptors

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    Jenkins Jeremy L

    2001-10-01

    Full Text Available Abstract Background To better understand the molecular interactions of Bt toxins with non-target insects, we have examined the real-time binding specificity and affinity of Cry1 toxins to native silkworm (Bombyx mori midgut receptors. Previous studies on B. mori receptors utilized brush border membrane vesicles or purifed receptors in blot-type assays. Results The Bombyx mori (silkworm aminopeptidase N (APN and cadherin-like receptors for Bacillus thuringiensis insecticidal Cry1Aa toxin were purified and their real-time binding affinities for Cry toxins were examined by surface plasmon resonance. Cry1Ab and Cry1Ac toxins did not bind to the immobilized native receptors, correlating with their low toxicities. Cry1Aa displayed moderate affinity for B. mori APN (75 nM, and unusually tight binding to the cadherin-like receptor (2.6 nM, which results from slow dissociation rates. The binding of a hybrid toxin (Aa/Aa/Ac was identical to Cry1Aa. Conclusions These results indicate domain II of Cry1Aa is essential for binding to native B. mori receptors and for toxicity. Moreover, the high-affinity binding of Cry1Aa to native cadherin-like receptor emphasizes the importance of this receptor class for Bt toxin research.

  16. The nicotinic acetylcholine receptor gene family of the silkworm, Bombyx mori

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    Zhang Chuan-Xi

    2007-09-01

    Full Text Available Abstract Background Nicotinic acetylcholine receptors (nAChRs mediate fast synaptic cholinergic transmission in the insect central nervous system. The insect nAChR is the molecular target of a class of insecticides, neonicotinoids. Like mammalian nAChRs, insect nAChRs are considered to be made up of five subunits, coded by homologous genes belonging to the same family. The nAChR subunit genes of Drosophila melanogaster, Apis mellifera and Anopheles gambiae have been cloned previously based on their genome sequences. The silkworm Bombyx mori is a model insect of Lepidoptera, among which are many agricultural pests. Identification and characterization of B. mori nAChR genes could provide valuable basic information for this important family of receptor genes and for the study of the molecular mechanisms of neonicotinoid action and resistance. Results We searched the genome sequence database of B. mori with the fruit fly and honeybee nAChRs by tBlastn and cloned all putative silkworm nAChR cDNAs by reverse transcriptase-polymerase chain reaction (RT-PCR and rapid amplification of cDNA ends (RACE methods. B. mori appears to have the largest known insect nAChR gene family to date, including nine α-type subunits and three β-type subunits. The silkworm possesses three genes having low identity with others, including one α and two β subunits, α9, β2 and β3. Like the fruit fly and honeybee counterparts, silkworm nAChR gene α6 has RNA-editing sites, and α4, α6 and α8 undergo alternative splicing. In particular, alternative exon 7 of Bmα8 may have arisen from a recent duplication event. Truncated transcripts were found for Bmα4 and Bmα5. Conclusion B. mori possesses a largest known insect nAChR gene family characterized to date, including nine α-type subunits and three β-type subunits. RNA-editing, alternative splicing and truncated transcripts were found in several subunit genes, which might enhance the diversity of the gene family.

  17. Partial deletions of the W chromosome due to reciprocal translocation in the silkworm Bombyx mori.

    Science.gov (United States)

    Abe, H; Seki, M; Ohbayashi, F; Tanaka, N; Yamashita, J; Fujii, T; Yokoyama, T; Takahashi, M; Banno, Y; Sahara, K; Yoshido, A; Ihara, J; Yasukochi, Y; Mita, K; Ajimura, M; Suzuki, M G; Oshiki, T; Shimada, T

    2005-08-01

    In the silkworm, Bombyx mori (female, ZW; male, ZZ), femaleness is determined by the presence of a single W chromosome, irrespective of the number of autosomes or Z chromosomes. The W chromosome is devoid of functional genes, except the putative female-determining gene (Fem). However, there are strains in which chromosomal fragments containing autosomal markers have been translocated on to W. In this study, we analysed the W chromosomal regions of the Zebra-W strain (T(W;3)Ze chromosome) and the Black-egg-W strain (T(W;10)+(w-2) chromosome) at the molecular level. Initially, we undertook a project to identify W-specific RAPD markers, in addition to the three already established W-specific RAPD markers (W-Kabuki, W-Samurai and W-Kamikaze). Following the screening of 3648 arbitrary 10-mer primers, we obtained nine W-specific RAPD marker sequences (W-Bonsai, W-Mikan, W-Musashi, W-Rikishi, W-Sakura, W-Sasuke, W-Yukemuri-L, W-Yukemuri-S and BMC1-Kabuki), almost all of which contained the border regions of retrotransposons, namely portions of nested retrotransposons. We confirmed the presence of eleven out of twelve W-specific RAPD markers in the normal W chromosomes of twenty-five silkworm strains maintained in Japan. These results indicate that the W chromosomes of the strains in Japan are almost identical in type. The Zebra-W strain (T(W;3)Ze chromosome) lacked the W-Samurai and W-Mikan RAPD markers and the Black-egg-W strain (T(W;10)+(w-2) chromosome) lacked the W-Mikan RAPD marker. These results strongly indicate that the regions containing the W-Samurai and W-Mikan RAPD markers or the W-Mikan RAPD marker were deleted in the T(W;3)Ze and T(W;10)+(w-2) chromosomes, respectively, due to reciprocal translocation between the W chromosome and the autosome. This deletion apparently does not affect the expression of Fem; therefore, this deleted region of the W chromosome does not contain the putative Fem gene.

  18. Identification and characterization of two putative microRNAs encoded by Bombyx mori cypovirus.

    Science.gov (United States)

    Pan, Zhong-Hua; Wu, Ping; Gao, Kun; Hou, Cheng-Xiang; Qin, Guang-Xing; Geng, Tao; Guo, Xi-Jie

    2017-04-02

    Viral microRNAs (miRNAs) have been demonstrated to play important roles in virus-host interactions. Some RNA virus-encoded miRNAs have been reported to promote viral replication and may be used as potential drug targets. Bombyx mori cypovirus (BmCPV), an important pathogen of silkworm, is a double-stranded RNA virus frequently causing serious damages in sericulture. Research on miRNA encoded by BmCPV may be useful to elucidate the BmCPV-host interaction and to develop new anti-viral methods. In our previous study, small RNA libraries of the midgut of BmCPV-infected silkworm have been generated by deep sequencing and several BmCPV-encoded putative miRNAs were predicted. In this study, two putative miRNAs encoded by BmCPV were identified and then validated by stem-loop qRT-PCR and northern blot. They are BmCPV-miR-3 encoded by the third genomic RNA segment of BmCPV (478-497bp) and BmCPV-miR-5 encoded by the fifth genomic RNA segment (2481-2500bp), both are 20bp and encoded by ORF regions. miRNA expression could be detected as early as 5h after BmCPV infection, and the expression level of BmCPV-miR-3 is much higher than that of BmCPV-miR-5 in the course of infection. Three potential target genes were predicted in the host genome, two for BmCPV-miR-3 and one for BmCPV-miR-5, but just one in the virus genome for BmCPV-miR-3 only, with the binding sites all in coding regions. Dual luciferase assay and qRT-PCR indicated that BmCPV-miR-3 could down-regulate the expression of both its two target genes, but no regulatory effect by BmCPV-miR-5 on its target gene was detected. In contrast, BmCPV-miR-3 could up-regulate the viral target. This is the first report that an insect double stranded RNA virus may generate miRNAs and the results obtained will benefit the future study of the functions of BmCPV-encoded miRNAs on viral replication and virus-host interaction. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. iTRAQ-based quantitative proteomic analysis of midgut in silkworm infected with Bombyx mori cytoplasmic polyhedrosis virus.

    Science.gov (United States)

    Gao, Kun; Deng, Xiang-Yuan; Shang, Meng-Ke; Qin, Guang-Xing; Hou, Cheng-Xiang; Guo, Xi-Jie

    2017-01-30

    Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) specifically infects the epithelial cells in the midgut of silkworm and causes them to death, which negatively affects the sericulture industry. In order to determine the midgut response at the protein levels to the virus infection, differential proteomes of the silkworm midgut responsive to BmCPV infection were identified with isobaric tags for relative and absolute quantitation (iTRAQ) labeling followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). 193, 408, 189 differentially expressed proteins (DEPs) were reliably quantified by iTRAQ analysis in the midgut of BmCPV-infected and control larvae at 24, 48, 72h post infection (hpi) respectively. KEGG enrichment analysis showed that Oxidative phosphorylation, amyotrophic lateral sclerosis, Toll-like receptor signaling pathway, steroid hormone biosynthesis were the significant pathways (Q value≤0.05) both at 24 and 48hpi. qRT-PCR was used to further verify gene transcription of 30 DEPs from iTRAQ, showing that the regulations of 24 genes at the transcript level were consistent with those at the proteomic level. Moreover, the cluster analysis of the three time groups showed that there were seven co-regulated DEPs including BGIBMGA002620-PA, which was a putative p62/sequestosome-1 protein in silkworm. It was upregulated at both the mRNA level and the proteomic level and may play an important role in regulating the autophagy and apoptosis (especially apoptosis) induced by BmCPV infection. This was the first report using an iTRAQ approach to analyze proteomes of the silkworm midgut against BmCPV infection, which contributes to understanding the defense mechanisms of silkworm midgut to virus infection. The domesticated silkworm, Bombyx mori, is renowned for silk production as well as being a traditional lepidopteron model insect served as a subject for morphological, genetic, physiological, and developmental studies. Bombyx mori cytoplasmic polyhedrosis

  20. Influência das dietas artificiais e folhas in natura no ganho de peso das larvas do bicho-da-seda (Bombyx mori L. Influence of artificial diets and natural leaves in weight gain of silkworm larvae (Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    Adriana Evangelista Rodrigues

    2000-05-01

    Full Text Available A utilização de dietas artificiais no Japão já é uma realidade e no Brasil iniciam-se os estudos, buscando encontrar um balanceamento que satisfaça as necessidades das larvas do bicho-da-seda (Bombyx mori L.. Desta forma, foram testados dois cultivares de amoreira como ingredientes das dietas, avaliando-se a qualidade do alimento através do ganho médio de peso das larvas. As larvas foram separadas em parcelas no início do 1º instar, quando já começaram a receber as dietas artificiais e as folhas in natura. Para o cálculo do ganho médio de peso analisou-se o peso inicial e final das larvas a cada instar. Quando as larvas receberam dietas artificiais no 1º e 2º instares e folhas in natura a partir do 3º instar não apresentaram diferenças significativas, sugerindo a utilização de dietas em criadeiras nas Fiações.The use of artificial diets in Japan is a well known fact. In Brazil experiments are assayed to find a balanced diet to meet the needs of the silkworm (Bombyx mori L.. Two mulberry cultivars were tested in diets to evaluate the quality of food by average gain of weight in larvae. Larvae were first separated in the first instar and fed with diets and natural leaves. As a rough estimate of mean gain of weight, initial and final weights in each instar were recorded. When the silkworms were fed with artificial diets in the first and second instars and with natural leaves in the third, fourth and fifth instars no significant differences were recorded. The use of diets in the first instars at the silk industry may be thus recommended.

  1. Alkaline phosphatase in the alimentary canal of silkworm larvae, Bombyx mori L. , poisoned by the oral application of fluorine compound

    Energy Technology Data Exchange (ETDEWEB)

    Fujii, M.

    1975-01-01

    Physiological damages caused by the oral administration of fluorine in Bombyx mori were investigated from a viewpoint of the enzyme activity of alimentary canal. The rate of hydrolysis of phosphate esters was determined using p-nitrophenolphosphate as a substrate. The enzyme activity in the midgut of treated silkworms always decreased, compared with that of the control larvae. The enzyme activities of the anterior segment of the midgut and the hind-gut were always low and no distinct variations were shown, while the enzyme activities of the middle and posterior segments for the midgut of untreated silkworms were high and those increased with growth. The increasing rates in body weight and enzyme activity were reduced in proportion to the fluorine concentration applied. Enzyme activity was not inhibited by the addition of fluoride in vitro, but rather activated by the addition of more than 10/sup 2/ ..mu..M NaF/s.5 ml (total).

  2. Effect of cocoon fluorescence, silkworm hybrid and gender on sericin content of Bombyx mori L. silk thread

    Directory of Open Access Journals (Sweden)

    M. Panayotov

    2016-06-01

    Full Text Available Abstract. The goal of the present study was to determine the influence of the ultraviolet fluorescence of cocoons, the hybrid, the sex and the interaction among them on the sericin content in silk threads. The study was performed with 3 di- and 2 tetra-cross silkmoth (Bombyx mori L. hybrids, differentiated in three groups – with violet, intermediate and yellow fluorescence of the cocoons. The examined factors had a significant effect (p≤0.001 on the sericin content. The highest sericin content was detected in the silk threads of the violet-fluorescent and the lowest – in the yellow-fluorescent group. The analysed di-hybrids were distinguished by better characteristics in terms of sericin content, compared to the tetra-hybrids, most obvious for the yellow-fluorescent fraction.

  3. Microscopy based studies on the interaction of bio-based silver nanoparticles with Bombyx mori Nuclear Polyhedrosis virus.

    Science.gov (United States)

    Tamilselvan, Selvaraj; Ashokkumar, Thirunavukkarasu; Govindaraju, Kasivelu

    2017-04-01

    In the present investigation, silver nanoparticles (AgNPs) interactions with Bombyx mori Nuclear Polyhedrosis virus (BmNPV) were characterized using High-Resolution Scanning Electron Microscopy (HR-SEM), Energy Dispersive X-ray Analysis (EDAX), Transmission Electron Microscopy (TEM), Atomic Force Microcopy (AFM) and Confocal Microscope (CM). HR-SEM study reveals that the biosynthesized AgNPs have interacted with BmNPV and were found on the surface. TEM micrographs of normal and viral polyhedra treated with AgNPs showed that the nanoparticles were accumulated in the membrane and it was noted that some of the AgNPs successfully penetrated the membrane by reaching the capsid of BmNPV. AFM and confocal microscopy studies reveal that the disruption in the shell membrane tends to lose its stability due to exposure of AgNPs to BmNPV. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Differentially expressed genes in the cuticle and hemolymph of the silkworm, Bombyx mori, injected with the fungus Beauveria bassiana.

    Science.gov (United States)

    Hou, Cheng-Xiang; Qin, Guang-Xing; Liu, Ting; Mei, Xing-Lin; Li, Bing; Shen, Zhong-Yuan; Guo, Xi-Jie

    2013-01-01

    The most important pathogenic fungus of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), is Beauveria bassiana (Balsamo-Crivelli ) Vuillemin (Hypocreales: Clavicipitaceae), which causes significant damage to sericulture production. Therefore, diagnosing fungal disease and developing new control measures are crucial to silk production. To better understand the responsive and interactive mechanisms between the host silkworm and this fungus, variations in silkworm gene expression were investigated using the suppression subtractive hybridization method following the injection of B. bassiana conidia. Two cDNA libraries were constructed, and 140 cDNA clones were isolated. Of the 50 differentially expressed genes identified, 45 (112 clones) were identified in the forward library, and 5 (28 clones) were identified in the reverse library. Expression profiling of six of these genes by quantitative polymerase chain reaction (qPCR) verified that they were induced by the fungal challenge. The present study provides insight into the interaction between lepidopteran insects and pathogenic fungi.

  5. Cerium chloride improves protein and carbohydrate metabolism of fifth-instar larvae of Bombyx mori under phoxim toxicity.

    Science.gov (United States)

    Li, Bing; Xie, Yi; Cheng, Zhe; Cheng, Jie; Hu, Rengping; Sang, Xuezi; Gui, Suxin; Sun, Qingqing; Gong, Xiaolan; Cui, Yaling; Shen, Weide; Hong, Fashui

    2012-12-01

    The organophosphorus pesticide poisoning of the silkworm Bombyx mori is one of the major events causing serious damage to sericulture. Added low-dose rare earths are demonstrated to increase resistance in animals. However, very little is known about whether or not added CeCl₃ can increase resistance of silkworm to phoxim poisoning. The present findings suggested that added CeCl₃ to mulberry leaves markedly increased contents of protein, glucose and pyruvate, and carbohydrate metabolism-related enzyme activities, including lactate dehydrogenase, succinate dehydrogenase and malate dehydrogenase, and attenuated free amino acids, urea, uric acid and lactate levels and inhibited the protein metabolism-related enzymes activities, such as protease, alanine aminotransferase and aspartate aminotransferase in the haemolymph of B. mori, under phoxim toxicity. These findings suggest that added CeCl₃ may improve protein and carbohydrate metabolisms, thus leading to increases of growth and survival rate of B. mori under phoxim stress.

  6. Juvenile Hormone Analogues, Methoprene and Fenoxycarb Dose-Dependently Enhance Certain Enzyme Activities in the Silkworm Bombyx Mori (L

    Directory of Open Access Journals (Sweden)

    M. Rajeswara Rao

    2008-06-01

    Full Text Available Use of Juvenile Hormone Analogues (JHA in sericulture practices has been shown to boost good cocoon yield; their effect has been determined to be dose-dependent. We studied the impact of low doses of JHA compounds such as methoprene and fenoxycarb on selected key enzymatic activities of the silkworm Bombyx mori. Methoprene and fenoxycarb at doses of 1.0 μg and 3.0fg/larvae/48 hours showed enhancement of the 5th instar B. mori larval muscle and silkgland protease, aspartate aminotransaminase (AAT and alanine aminotransaminase (ALAT, adenosine triphosphate synthase (ATPase and cytochrome-c-oxidase (CCO activity levels, indicating an upsurge in the overall oxidative metabolism of the B.mori larval tissues.

  7. Rapid Detection of Infectious Flacherie Virus of the Silkworm, Bombyx mori, using RT-PCR and Nested PCR

    Science.gov (United States)

    Vootla, Shyam Kumar; Lu, Xing Meng; Kari, Neetha; Gadwala, Mallikarjun; Lu, Qineng

    2013-01-01

    In this study, a method for detection of an ssRNA viral pathogen that causes viral flacherie in the silkworm, Bombyx mori (L.) (Lepidoptera: Bombycidae), was used for the detection of B. mori infectious flacherie virus (BmIFV). A combination of nested and reverse transcriptase polymerase chain reaction was used for detection. Although BmIFV has been reported in almost all the sericultural regions of the world, there had been no reports of BmIFV incidence in India. Therefore, the confirmation of the presence of BmIFV in Karnataka, India, is of great significance. The present method is advantageous because it can be used to detect the virus by using samples from infected midgut tissues, thus simplifying and avoiding laborious genome isolation procedures. This method could help in early detection of BmIFV disease pathogens and help reduce crop losses. PMID:24785655

  8. Juvenile Hormone Analogues, methoprene and fenoxycarb dose-dependently enhance certain enzyme activities in the silkworm Bombyx mori (L).

    Science.gov (United States)

    Mamatha, Devi M; Kanji, Vijaya K; Cohly, Hari H P; Rao, M Rajeswara

    2008-06-01

    Use of Juvenile Hormone Analogues (JHA) in sericulture practices has been shown to boost good cocoon yield; their effect has been determined to be dose-dependent. We studied the impact of low doses of JHA compounds such as methoprene and fenoxycarb on selected key enzymatic activities of the silkworm Bombyx mori. Methoprene and fenoxycarb at doses of 1.0 microg and 3.0 fg/larvae/48 hours showed enhancement of the 5th instar B. mori larval muscle and silkgland protease, aspartate aminotransaminase (AAT) and alanine aminotransaminase (ALAT), adenosine triphosphate synthase (ATPase) and cytochrome-c-oxidase (CCO) activity levels, indicating an upsurge in the overall oxidative metabolism of the B.mori larval tissues.

  9. Determination of sialic acids in the nervous system of silkworm (Bombyx mori L.: Effects of aging and development

    Directory of Open Access Journals (Sweden)

    Soya Seçkin

    2017-01-01

    Full Text Available Sialic acids mainly occur as components on cell surface glycoproteins and glycolipids. They play a major role in the chemical and biological diversity of glycoconjugates. Although sialic acids exhibit great structural variability in vertebrates, glycoconjugates with sialic acids have also been determined in small amounts in invertebrates. It has been suggested that sialic acids play important roles in the development and function of the nervous system. Despite Bombyx mori being a model organism for the investigation of many physiological processes, sialic acid changes in its nervous system have not been examined during development and aging. Therefore, in this study we aimed to determine sialic acid changes in the nervous system of Bombyx mori during development and aging processes. Liquid chromatography-mass spectrometry (LC-MS and lectin immunohistochemistry were carried out in order to find variations among different developmental stages. Developmental stages were selected as 3rd instar (the youngest and 5th larval instar (young, motionless prepupa (the oldest and 13-day-old pupa (adult development. At all stages, only Neu5Ac was present, however, it dramatically decreased during the developmental and aging stages. On the other hand, an increase was observed in the amount of Neu5Ac during the pupal stage. In immunohistochemistry experiments with Maackia amurensis agglutinin (MAA and Sambucus nigra agglutinin (SNA lectins, the obtained staining was consistent with the obtained LC-MS results. These findings indicate that sialic acids are abundant at the younger stages but that they decrease in the insect nervous system during development and aging, similarly as in mammals.

  10. A Hox Gene, Antennapedia, Regulates Expression of Multiple Major Silk Protein Genes in the Silkworm Bombyx mori*

    Science.gov (United States)

    Tsubota, Takuya; Tomita, Shuichiro; Uchino, Keiro; Kimoto, Mai; Takiya, Shigeharu; Kajiwara, Hideyuki; Yamazaki, Toshimasa; Sezutsu, Hideki

    2016-01-01

    Hox genes play a pivotal role in the determination of anteroposterior axis specificity during bilaterian animal development. They do so by acting as a master control and regulating the expression of genes important for development. Recently, however, we showed that Hox genes can also function in terminally differentiated tissue of the lepidopteran Bombyx mori. In this species, Antennapedia (Antp) regulates expression of sericin-1, a major silk protein gene, in the silk gland. Here, we investigated whether Antp can regulate expression of multiple genes in this tissue. By means of proteomic, RT-PCR, and in situ hybridization analyses, we demonstrate that misexpression of Antp in the posterior silk gland induced ectopic expression of major silk protein genes such as sericin-3, fhxh4, and fhxh5. These genes are normally expressed specifically in the middle silk gland as is Antp. Therefore, the evidence strongly suggests that Antp activates these silk protein genes in the middle silk gland. The putative sericin-1 activator complex (middle silk gland-intermolt-specific complex) can bind to the upstream regions of these genes, suggesting that Antp directly activates their expression. We also found that the pattern of gene expression was well conserved between B. mori and the wild species Bombyx mandarina, indicating that the gene regulation mechanism identified here is an evolutionarily conserved mechanism and not an artifact of the domestication of B. mori. We suggest that Hox genes have a role as a master control in terminally differentiated tissues, possibly acting as a primary regulator for a range of physiological processes. PMID:26814126

  11. Effects of Titanium Dioxide Nanoparticles on the Synthesis of Fibroin in Silkworm (Bombyx mori).

    Science.gov (United States)

    Ni, Min; Li, FanChi; Tian, JiangHai; Hu, JingSheng; Zhang, Hua; Xu, KaiZun; Wang, BinBin; Li, YangYang; Shen, WeiDe; Li, Bing

    2015-08-01

    Silkworm (Bombyx mori) is an economically important insect, and its silk production capacity largely depends on its ability to synthesize fibroin. While breeding of B. mori varieties has been a key strategy to improve silk production, little improvement of B. mori silk production has been achieved to date. As a result, the development of sericulture economy has not progressed well, pointing to the need of new ways for improvement of B. mori silk production. Titanium dioxide nanoparticles (TiO2 NPs), a food additive widely used for livestock, have been shown to promote animal growth and increase the protein synthesis in animals. However, no studies on effect of TiO2 NPs on fibroin synthesis in B. mori have been available. In this study, the differential expression profiles of genes and proteins in the silk gland of B. mori fed without or with TiO2 NPs (5 μg ml(-1)) were analyzed and compared using digital gene expression (DGE), reverse transcription quantitative polymerase chain reaction (RT-qPCR), semi-qPCR, and Western blot analysis. The effects of TiO2 NPs feeding on the activity of proteases in the midgut and the synthesis and transportation of amino acids in hemolymph were also investigated. DGE analyses showed that among a total of 4,741 genes detected, 306 genes were differentially expressed after the TiO2 NPs feeding, of which 137 genes were upregulated whereas 169 genes were downregulated. 106 genes were shown to be involved in fibroin synthesis, of which 97 genes, including those encoding cuticular protein glycine-rich 10, serine protease inhibitor 28, aspartate aminotransferase, lysyl-tRNA synthetase, and splicing factor arginine/serine-rich 6, and silk gland factor-1 (SGF-1), were upregulated with the maximum induction of 8.52-folds, whereas nine genes, including those encoding aspartylglucosaminidase, the cathepsin L in Tribolium castaneum, and similar to SPRY domain-containing SOCS box protein 3, were downregulated with the maximum reduction of 8

  12. Comparative Study of Biological Characteristics of Larvae, Crude and Dried Cocoon in 7 Races of Silkworm Bombyx mori L., Raised in Transylvania Area

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    Emilia Maria Furdui

    2010-05-01

    Full Text Available 7 monovoltine races of Romanian silkworm Bombyx mori L. were breeded in specific Transylvania conditions, assuring the same microclimate conditions, being fed with the same mulberry leaves (Ukraina 107. The results obtained showed a high homogeneity, the differences were due to the variability and genotype characteristic for each individual of every race. Results and standard deviations are within the race standard. The study permitted a ierarchy of the races, following the biological characteristics, microclimate conditions, quantity, quality and breeding technology.

  13. Efisiensi Konsumsi Pakan Dan Laju Respirasi Ulat Sutera Bombyx mori L. (Lepidoptera: Bombicidae) Yang Diberi Daun Murbei (Morus sp.) Yang Mengandung Vitamin B1 (TIAMIN)

    OpenAIRE

    Hayani, Rizma

    2014-01-01

    The Effect of Mulberry (Morus sp.) Leave that Contain Vitamin B1 (Thiamine) on the Efficiency of Feed Consumption and Respiration Rate of Silkworm Bombyx mori L. (Lepidoptera: Bombicidae)” has been carried out in the Laboratory of Genetics, Faculty of Mathematics and Natural Sciences, University of Sumatra Utara, Medan. This study used Complete Random Design (CRD) with 5 treatments and 3 replications each replication consisted of 10 silkworm. The treatmants that vitamin B1 concentration of mg...

  14. Resistance to BmNPV via overexpression of an exogenous gene controlled by an inducible promoter and enhancer in transgenic silkworm, Bombyx mori.

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    Liang Jiang

    Full Text Available The hycu-ep32 gene of Hyphantria cunea NPV can inhibit Bombyx mori nucleopolyhedrovirus (BmNPV multiplication in co-infected cells, but it is not known whether the overexpression of the hycu-ep32 gene has an antiviral effect in the silkworm, Bombyx mori. Thus, we constructed four transgenic vectors, which were under the control of the 39 K promoter of BmNPV (39 KP, Bombyx mori A4 promoter (A4P, hr3 enhancer of BmNPV combined with 39 KP, and hr3 combined with A4P. Transgenic lines were created via embryo microinjection using practical diapause silkworm. qPCR revealed that the expression level of hycu-ep32 could be induced effectively after BmNPV infection in transgenic lines where hycu-ep32 was controlled by hr3 combined with 39 KP (i.e., HEKG. After oral inoculation of BmNPV with 3 × 10(5 occlusion bodies per third instar, the mortality with HEKG-B was approximately 30% lower compared with the non-transgenic line. The economic characteristics of the transgenic lines remained unchanged. These results suggest that overexpression of an exogenous antiviral gene controlled by an inducible promoter and enhancer is a feasible method for breeding silkworms with a high antiviral capacity.

  15. Resistance to BmNPV via Overexpression of an Exogenous Gene Controlled by an Inducible Promoter and Enhancer in Transgenic Silkworm, Bombyx mori

    Science.gov (United States)

    Jiang, Liang; Cheng, Tingcai; Zhao, Ping; Yang, Qiong; Wang, Genhong; Jin, Shengkai; Lin, Ping; Xiao, Yang; Xia, Qingyou

    2012-01-01

    The hycu-ep32 gene of Hyphantria cunea NPV can inhibit Bombyx mori nucleopolyhedrovirus (BmNPV) multiplication in co-infected cells, but it is not known whether the overexpression of the hycu-ep32 gene has an antiviral effect in the silkworm, Bombyx mori. Thus, we constructed four transgenic vectors, which were under the control of the 39 K promoter of BmNPV (39 KP), Bombyx mori A4 promoter (A4P), hr3 enhancer of BmNPV combined with 39 KP, and hr3 combined with A4P. Transgenic lines were created via embryo microinjection using practical diapause silkworm. qPCR revealed that the expression level of hycu-ep32 could be induced effectively after BmNPV infection in transgenic lines where hycu-ep32 was controlled by hr3 combined with 39 KP (i.e., HEKG). After oral inoculation of BmNPV with 3 × 105 occlusion bodies per third instar, the mortality with HEKG-B was approximately 30% lower compared with the non-transgenic line. The economic characteristics of the transgenic lines remained unchanged. These results suggest that overexpression of an exogenous antiviral gene controlled by an inducible promoter and enhancer is a feasible method for breeding silkworms with a high antiviral capacity. PMID:22870254

  16. The angiotensin converting enzyme (ACE) inhibitor, captopril disrupts the motility activation of sperm from the silkworm, Bombyx mori.

    Science.gov (United States)

    Nagaoka, Sumiharu; Kawasaki, Saori; Kawasaki, Hideki; Kamei, Kaeko

    2017-11-01

    Angiotensin I-converting enzyme (also known as peptidyl dicarboxypeptidase A, ACE, and EC 3.4.15.1), which is found in a wide range of organisms, cleaves C-terminal dipeptides from relatively short oligopeptides. Mammalian ACE plays an important role in the regulation of blood pressure. However, the precise physiological functions of insect ACE homologs have not been understood. As part of our effort to elucidate new physiological roles of insect ACE, we herein report a soluble ACE protein in male reproductive secretions from the silkmoth, Bombyx mori. Seminal vesicle sperm are quiescent in vitro, but vigorous motility is activated by treatment with either a glandula (g.) prostatica homogenate or trypsin in vitro. When seminal vesicle sperm were pre-incubated with captopril, a strong and specific inhibitor of mammalian ACE, and then stimulated to initiate motility by the addition of the g. prostatica homogenate or trypsin, the overall level of acquired motility was reduced in an inhibitor-concentration-dependent manner. In the course of this project, we detected ACE-related carboxypeptidase activity that was inhibited by captopril in both the vesicular (v.) seminalis of the noncopulative male reproductive tract and in the spermatophore that forms in the female bursa copulatrix at the time of mating, just as in an earlier report on the tomato moth, Lacanobia oleracea, which belongs to a different lepidopteran species (Ekbote et al., 2003a). Two distinct genes encoding ACE-like proteins were identified by analysis of B. mori cDNA, and were named BmAcer and BmAcer2, respectively [the former was previously reported by Quan et al. (2001) and the latter was first isolated in this paper]. RT-qPCR and Western blot analyses indicated that the BmAcer2 was predominantly produced in v. seminalis and transferred to the spermatophore during copulation, while the BmAcer was not detected in the adult male reproductive organs. A recombinant protein of BmAcer2 (devoid of a signal

  17. Gamma radiation effects of {sup 60} Co on Bombyx mori (Lep., Bombycidae) modifying the silk fiber production; Influencia da radiacao gama ({sup 60} Co) na producao de fios de seda em Bombyx mori(Lep.,Bombycidae)

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    Carneiro Junior, Francisco [Universidade Metodista de Piracicaba (UNIMEP), SP (Brazil); Bendassolli, Jose A. [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil)

    1997-12-01

    The present work aimed to verify the biological effects of the application of different doses of gamma radiation during the fifth instar of the silkworm catepillar. Sevently eight silkworm caterpillars (Bombyx mori) were irradiated with {gamma}{sup 60} Co radiation at the initial period of the fifth instar. The caterpillars were divided and classified in six batches of thirteen individuals each. Treatments 1 through 5 received 20, 40, 60, 80 and 100 Gy, respectively, and the control, also consisted of thirteen caterpillars, was not irradiated. The results showed a general increase in the silk fiber content in the irradiated batches compared to the control. The weight of the silk cocoons was higher with increasing doses of irradiation, from 20 to 80 Gy, respectively, followed by a decrease in weight in the treatment irradiated with 100 Gy. the results obtained in this experiment enable the conclusion that the radiation applied to the caterpillars significantly influenced the production of silk fiber in this species. (author). 4 refs., 2 figs., 3 tabs.

  18. Activation of BNGR-A24 by direct interaction with tachykinin-related peptides from the silkworm Bombyx mori leads to the Gq- and Gs-coupled signaling cascades.

    Science.gov (United States)

    He, Xiaobai; Zang, Jiashu; Li, Xiangmei; Shao, Jiajie; Yang, Huipeng; Yang, Jingwen; Huang, Haishan; Chen, Linjie; Shi, Liangen; Zhu, Chenggang; Zhang, Guozheng; Zhou, Naiming

    2014-10-28

    Tachykinins constitute one of the largest peptide families in the animal kingdom and exert their diverse actions via G protein-coupled receptors (GPCRs). In this study, the Bombyx tachykinin-related peptides (TKRPs) were identified as specific endogenous ligands for the Bombyx neuropeptide GPCR A24 (BNGR-A24) and thus designated BNGR-A24 as BmTKRPR. Using both mammalian cell line HEK293 and insect cell line Sf21, further characterization demonstrated that BmTKRPR was activated, thus resulting in intracellular accumulation of cAMP, Ca(2+) mobilization, and ERK1/2 phosphorylation in a Gs and Gq inhibitor-sensitive manner. Moreover, quantitative reverse transcriptase polymerase chain reaction analysis and dsRNA-mediated knockdown experiments suggested a possible role for BmTKRPR in the regulation of feeding and growth. Our findings enhance the understanding of the Bombyx TKRP system in the regulation of fundamental physiological processes.

  19. Molecular cloning and characterization of peroxiredoxin 4 involved in protection against oxidative stress in the silkworm Bombyx mori.

    Science.gov (United States)

    Shi, G-Q; Yu, Q-Y; Shi, L; Zhang, Z

    2012-12-01

    Peroxiredoxins (Prxs) are a ubiquitous family of proteins that play important roles in insects in protection against oxidative stress through the detoxification of cellular peroxides. Here, we describe the cloning and characterization of a Prx4 cDNA of the silkworm Bombyx mori (BmPrx4). The BmPrx4 gene has an open reading frame of 744 bp encoding 248 amino acids and a conserved motif, VCP, involved in its presumed redox functions. The heterologously expressed proteins of the gene in Escherichia coli showed antioxidant activity, removed hydrogen peroxide and protected DnA. Western blotting analysis showed the presence of BmPrx4 in the haemolymph, suggesting that the protein is secretable. Moreover, BmPrx4 was expressed at all developmental stages. The expression level of BmPrx4 was relatively low during the feeding stage but high at the wandering stage. BmPrx4 was induced by quercetin or temperature stress. Immunohistochemical analysis revealed that BmPrx4 is present in the brain, neurones and olfactory organ of the head in silkworms. Overall, our results indicate that the expression profile of BmPrx4 correlates well with protection from oxidative damage. Our data provide clues for the development of control technology for agricultural and forestry pests as the silkworm is a representative of lepidopteran pests. © 2012 Royal Entomological Society.

  20. Preparation of Porous Scaffolds from Silk Fibroin Extracted from the Silk Gland of Bombyx mori (B. mori

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    Liangjun Zhu

    2012-06-01

    Full Text Available In order to use a simple and ecofriendly method to prepare porous silk scaffolds, aqueous silk fibroin solution (ASF was extracted from silk gland of 7-day-old fifth instar larvae of Bombyx mori (B. mori. SDS-page analysis indicated that the obtained fibroin had a molecular weight higher than 200 kDa. The fabrication of porous scaffolds from ASF was achieved by using the freeze-drying method. The pore of porous scaffolds is homogenous and tends to become smaller with an increase in the concentration of ASF. Conversely, the porosity is decreased. The porous scaffolds show impressive compressive strength which can be as high as 6.9 ± 0.4 MPa. Furthermore, ASF has high cell adhesion and growth activity. It also exhibits high ALP activity. This implies that porous scaffolds prepared from ASF have biocompatibility. Therefore, the porous scaffolds prepared in this study have potential application in tissue engineering due to the impressive compressive strength and biocompatibility.

  1. Respective Functions of Two Distinct Siwi Complexes Assembled during PIWI-Interacting RNA Biogenesis in Bombyx Germ Cells

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    Kazumichi M. Nishida

    2015-01-01

    Full Text Available PIWI-interacting RNA (piRNA biogenesis consists of two sequential steps: primary piRNA processing and the ping-pong cycle that depends on reciprocal Slicer-mediated RNA cleavage by PIWI proteins. However, the molecular functions of the factors involved remain elusive. Here, we show that RNAs cleaved by a Bombyx mori PIWI, Siwi, remain bound to the protein upon cleavage but are released by a DEAD box protein BmVasa. BmVasa copurifies with Siwi but not another PIWI BmAgo3. A lack of BmVasa does not affect primary piRNA processing but abolishes the ping-pong cycle. Siwi also forms a complex with BmSpn-E and BmQin. This complex is physically separable from the Siwi/BmVasa complex. BmSpn-E, unlike BmVasa, is necessary for primary piRNA production. We propose a model for piRNA biogenesis, where the BmSpn-E/BmQin dimer binds Siwi to function in primary piRNA processing, whereas BmVasa, by associating with Siwi, ensures target RNA release upon cleavage to facilitate the ping-pong cycle.

  2. Novel female-specific trans-spliced and alternative splice forms of dsx in the silkworm Bombyx mori.

    Science.gov (United States)

    Duan, Jianping; Xu, Hanfu; Wang, Feng; Ma, Sanyuan; Zha, Xingfu; Guo, Huizhen; Zhao, Ping; Xia, Qingyou

    2013-02-15

    The Bombyx mori doublesex gene (Bmdsx) plays an important role in somatic sexual development. Its pre-mRNA splices in a sex-specific manner to generate two female-specific and one male-specific splice forms. The present study investigated six novel dsx variants generated by trans-splicing between female dsx transcripts and two additional novel genes, dsr1 and dsr2. Expression analysis indicated that Bmdsx-dsr1 represented splicing noise, whereas dsr2, which trans-spliced with dsx to generate five variants, regulated the expression of the female-specific B. mori dsx transcript Bmdsx(F)s. We unexpectedly found a novel exon 2n insertion during Bmdsx transcription, which did not influence the validity of the novel protein, BmDSX(F3). Ectopic expression of BmDSX(F3) repressed the pheromone-binding protein gene and the testis-specific gene A2 in males, and activated of the storage protein 1 gene. Our findings suggest that trans-splicing is a novel regulatory function of Bmdsx, which participates in female sexual development by regulating the expression of three BmDSX(F) proteins. Copyright © 2012 Elsevier Inc. All rights reserved.

  3. Development of a foot-and-mouth disease virus serotype A empty capsid subunit vaccine using silkworm (Bombyx mori pupae.

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    Zhiyong Li

    Full Text Available Foot-and-mouth disease (FMD is a highly contagious disease of cloven-hoofed animals that inflicts severe economic losses in the livestock industry. In 2009, FMDV serotype A caused outbreaks of FMD in cattle in China. Although an inactivated virus vaccine has proven effective to control FMD, its use may lead to new disease outbreaks due to a possible incomplete inactivation of the virus during the manufacturing process. Here, we expressed the P1-2A and the 3C coding regions of a serotype A FMDV field isolate in silkworm pupae (Bombyx mori and evaluated the immunogenicity of the expression products. Four of five cattle vaccinated with these proteins developed high titers of FMDV-specific antibody and were completely protected against virulent homologous virus challenge with 10,000 50% bovine infectious doses (BID(50. Furthermore, the 50% bovine protective dose (PD(50 test was performed to assess the bovine potency of the empty capsid subunit vaccine and was shown to achieve 4.33 PD(50 per dose. These data provide evidence that silkworm pupae can be used to express immunogenic FMDV proteins. This strategy might be used to develop a new generation of empty capsid subunit vaccines against a variety of diseases.

  4. 20-Hydroxyecdysone (20E) Primary Response Gene E75 Isoforms Mediate Steroidogenesis Autoregulation and Regulate Developmental Timing in Bombyx*

    Science.gov (United States)

    Li, Kang; Tian, Ling; Guo, Zhongjian; Guo, Sanyou; Zhang, Jianzhen; Gu, Shi-Hong; Palli, Subba R.; Cao, Yang; Li, Sheng

    2016-01-01

    The temporal control mechanisms that precisely control animal development remain largely elusive. The timing of major developmental transitions in insects, including molting and metamorphosis, is coordinated by the steroid hormone 20-hydroxyecdysone (20E). 20E involves feedback loops to maintain pulses of ecdysteroid biosynthesis leading to its upsurge, whereas the underpinning molecular mechanisms are not well understood. Using the silkworm Bombyx mori as a model, we demonstrated that E75, the 20E primary response gene, mediates a regulatory loop between ecdysteroid biosynthesis and 20E signaling. E75 isoforms A and C directly bind to retinoic acid receptor-related response elements in Halloween gene promoter regions to induce gene expression thus promoting ecdysteroid biosynthesis and developmental transition, whereas isoform B antagonizes the transcriptional activity of isoform A/C through physical interaction. As the expression of E75 isoforms is differentially induced by 20E, the E75-mediated regulatory loop represents a fine autoregulation of steroidogenesis, which contributes to the precise control of developmental timing. PMID:27365399

  5. The effects of Bombyx mori silk strain and extraction time on the molecular and biological characteristics of sericin.

    Science.gov (United States)

    Siritientong, Tippawan; Bonani, Walter; Motta, Antonella; Migliaresi, Claudio; Aramwit, Pornanong

    2016-01-01

    Sericin was extracted from three strains of Thai Bombyx mori silk cocoons (white shell Chul1/1, greenish shell Chul3/2, and yellow shell Chul4/2) by a high-pressure and high-temperature technique. The characteristics of sericin extracted from different fractions (15, 45, and 60 min extraction process) were compared. No differences in amino acid composition were observed among the three fractions. For all silk strains, sericin extracted from a 15-min process presented the highest molecular weight. The biological potential of the different sericin samples as a bioadditive for 3T3 fibroblast cells was assessed. When comparing sericin extracted from three silk strains, sericin fractions extracted from Chul4/2 improved cell proliferation, while sericin from Chul 1/1 activated Type I collagen production to the highest extent. This study allows the natural variability of sericin obtained from different sources and extraction conditions to be addressed and provides clues for the selection of sericin sources.

  6. Comparative proteomics reveal diverse functions and dynamic changes of Bombyx mori silk proteins spun from different development stages.

    Science.gov (United States)

    Dong, Zhaoming; Zhao, Ping; Wang, Chen; Zhang, Yan; Chen, Jianping; Wang, Xin; Lin, Ying; Xia, Qingyou

    2013-11-01

    Silkworms (Bombyx mori) produce massive amounts of silk proteins to make cocoons during the final stages of larval development. Although the major components, fibroin and sericin, have been the focus for a long time, few researchers have realized the complexity of the silk proteome. We collected seven kinds of silk fibers spun by silkworm larvae at different developmental stages: the silks spun by new hatched larvae, second instar day 0 larvae, third instar day 0 larvae, fourth instar day 0 larvae, and fourth instar molting larvae, the scaffold silk used to attach the cocoon to the substrate and the cocoon silk. Analysis by liquid chromatography-tandem mass spectrometry identified 500 proteins from the seven silks. In addition to the expected fibroins, sericins, and some known protease inhibitors, we also identified further protease inhibitors, enzymes, proteins of unknown function, and other proteins. Unsurprisingly, our quantitative results showed fibroins and sericins were the most abundant proteins in all seven silks. Except for fibroins and sericins, protease inhibitors, enzymes, and proteins of unknown function were more abundant than other proteins. We found significant change in silk protein compositions through development, being consistent with their different biological functions and complicated formation.

  7. Shotgun proteomics approach to characterizing the embryonic proteome of the silkworm, Bombyx mori, at labrum appearance stage.

    Science.gov (United States)

    Li, J-Y; Chen, X; Hosseini Moghaddam, S H; Chen, M; Wei, H; Zhong, B-X

    2009-10-01

    The shotgun approach has gained considerable acknowledgement in recent years as a dominant strategy in proteomics. We observed a dramatic increase of specific protein spots in two-dimensional electrophoresis (2-DE) gels of the silkworm (Bombyx mori) embryo at labrum appearance, a characteristic stage during embryonic development of silkworm which is involved with temperature increase by silkworm raiser. We employed shotgun liquid chromatography tandem mass spectrometry (LC-MS/MS) technology to analyse the proteome of B. mori embryos at this stage. A total of 2168 proteins were identified with an in-house database. Approximately 47% of them had isoelectric point (pI) values distributed theoretically in the range pI 5-7 and approximately 60% of them had molecular weights of 15-45 kDa. Furthermore, 111 proteins had an pI greater than 10 and were difficult to separate by 2-DE. Many important functional proteins related to embryonic development, stress response, DNA transcription/translation, cell growth, proliferation and differentiation, organogenesis and reproduction were identified. Among them proteins related to nervous system development were noticeable. All known heat shock proteins (HSPs) were detected in this developmental stage of B. mori embryo. In addition, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed energetic metabolism at this stage. These results were expected to provide more information for proteomic monitoring of the insect embryo and better understanding of the spatiotemporal expression of genes during embryonic developmental processes.

  8. Studies on the chitin/chitosan binding properties of six cuticular proteins analogous to peritrophin 3 from Bombyx mori.

    Science.gov (United States)

    Qu, M; Ren, Y; Liu, Y; Yang, Q

    2017-08-01

    Chitin deacetylation is required to make the cuticle rigid and compact through chitin chain crosslinking. Thus it is presumed that specialized proteins are required to bind deacetylated chitin chains together. However, deacetylated-chitin binding proteins have not ever been reported. In a previous work, six cuticular proteins analogous to peritrophin 3 (CPAP3s) were found to be abundant in the moulting fluid of Bombyx mori. In this study, these BmCPAP3s (BmCPAP3-A1, BmCPAP3-A2, BmCPAP3-B, BmCPAP3-C, BmCPAP3-D1 and BmCPAP3-D2) were cloned and expressed in Escherichia coli and purified using metal-chelating affinity chromatography. Their binding activities demonstrated that although all of the BmCPAP3s showed similar binding abilities toward crystalline chitin and colloidal chitin, they differed in their affinities toward partially and fully deacetylated chitin. Amongst them, BmCPAP3-D1 exhibited the highest binding activity toward deacetylated chitin. The gene expression pattern of BmCPAP3-D1 was similar to BmCPAP3-A1 and BmCPAP3-C at most stages except that it was dramatically upregulated at the beginning of the pupa to adult transition stage. This work is the first report of a chitin-binding protein, BmCPAP3-D1, which exhibits high binding affinity to deacetylated chitin. © 2017 The Royal Entomological Society.

  9. Identification and Characterization of Novel Chitin-Binding Proteins from the Larval Cuticle of Silkworm, Bombyx mori.

    Science.gov (United States)

    Dong, Zhaoming; Zhang, Weiwei; Zhang, Yan; Zhang, Xiaolu; Zhao, Ping; Xia, Qingyou

    2016-05-06

    Cuticle is mainly made of chitin filaments embedded in a matrix of cuticular proteins (CPs). Cuticular chitins have minor differences, whereas CPs are widely variable with respect to their sequences and structures. To understand the molecular basis underlying the mechanical properties of cuticle, it is necessary to know which CPs interact with chitin and how they are assembled into the cuticle structure. In the present study, a chitin-binding assay was performed followed by liquid chromatography-tandem mass spectrometry to identify the extracted proteins from the larval cuticle of silkworm, Bombyx mori. There were 463 proteins identified from the silkworm larval cuticle, 200 of which were recovered in the chitin-binding fraction. A total of 103 proteins were annotated as CPs, which were classified into 11 CP families based on their conserved motifs, including CPR, CPAP, CPT, CPF and CPFL, CPCFC, chitin_bind 3, BmCPH2 homologues, BmCPH9 homologues, BmCPG1 homologues, BmCPG20 homologues, and BmCPG21 homologues. A total of five CP families were newly identified in the chitin-binding fraction, thereby providing new information and insight into the composition, structure, and function of the silkworm larval cuticle.

  10. Bm59 is an early gene, but is unessential for the propagation and assembly of Bombyx mori nucleopolyhedrovirus.

    Science.gov (United States)

    Hu, Xiaolong; Shen, Yunwang; Zheng, Qin; Wang, Guobao; Wu, Xiaofeng; Gong, Chengliang

    2016-02-01

    Bombyx mori nucleopolyhedrovirus (BmNPV) is a major pathogen that specifically infects the domestic silkworm and causes serious economic loss to sericulture around the world. The function of BmNPV Bm59 gene in the viral life cycle is inconclusive. To investigate the role of Bm59 during viral infection, the transcription initiation site and temporal expression of Bm59 were analyzed, and Bm59-knockout virus was generated through homologous recombination in Escherichia coli. The results showed that Bm59 is an early transcription gene with an atypia early transcriptional start motif. Budded virion (BV) production and DNA replication in the BmN cells transfected with the Bm59-knockout virus bacmid were similar to those in the cells transfected with the wild-type virus. Electron microscopy revealed that the occlusion-derived virus can be produced in cells infected with the Bm59-knockout virus. These results indicated that Bm59 is an early gene and is not essential for viral replication or assembly of BmNPV. These findings suggested that non-essential gene (Bm59) remained in the viral genome, which may interact with other viral/host genes in a certain situation.

  11. dsRNA interference on expression of a RNA-dependent RNA polymerase gene of Bombyx mori cytoplasmic polyhedrosis virus.

    Science.gov (United States)

    Pan, Zhong-Hua; Gao, Kun; Hou, Cheng-Xiang; Wu, Ping; Qin, Guang-Xing; Geng, Tao; Guo, Xi-Jie

    2015-07-01

    Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) is one of the major viral pathogens in silkworm. Its infection often results in significant losses to sericulture. Studies have demonstrated that RNAi is one of the important anti-viral mechanisms in organisms. In this study, three dsRNAs targeting the RNA-dependent RNA polymerase (RDRP) gene of BmCPV were designed and synthesized with 2'-F modification to explore their interference effects on BmCPV replication in silkworm larvae. The results showed that injecting dsRNA in the dosage of 4-6 ng per mg body weight into the 5th instar larvae can interfere with the BmCPV-RDRP expression by 93% after virus infection and by 99.9% before virus infection. In addition, the expression of two viral structural protein genes (genome RNA segments 1 and 5) was also decreased with the decrease of RDRP expression, suggesting that RNAi interference of BmCPV-RDRP expression could affect viral replication. The study provides an effective method for investigating virus replication as well as the virus-host interactions in the silkworm larvae using dsRNA. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Abortive replication of Bombyx mori nucleopolyhedrovirus in Sf9 and High Five cells: Defective nuclear transport of the virions

    International Nuclear Information System (INIS)

    Katou, Yasuhiro; Ikeda, Motoko; Kobayashi, Michihiro

    2006-01-01

    Despite close genetic relationship, Bombyx mori nucleopolyhedrovirus (BmNPV) and Autographa californica multicapsid NPV (AcMNPV) display a distinct host range property. Here, BmNPV replication was examined in Sf9 and High Five cells that were nonproductive for BmNPV infection but supported high titers of AcMNPV replication. Recombinant BmNPV, vBm/gfp/lac, containing bm-ie1 promoter-driven egfp showed that few Sf9 and High Five cells infected with vBm/gfp/lac expressed EGFP, while large proportion of EGFP-expressing cells was observed when transfected with vBm/gfp/lac DNA. Immunocytochemical analysis showed that BmNPV was not imported into the nucleus of these two cell lines, while recombinant BmNPV, vBmΔ64/ac-gp64 possessing AcMNPV gp64 was imported into the nucleus, yielding progeny virions in High Five cells, but not Sf9 cells. These results indicate that the defective nuclear import of infected virions due to insufficient BmNPV GP64 function is involved in the restricted BmNPV replication in Sf9 and High Five cells

  13. The sexual difference in degradation of sulfur amino acids in pupae and adults of the silkworm, Bombyx mori

    International Nuclear Information System (INIS)

    Shinbo, Hiroshi

    1978-01-01

    The degradation of sulfur amino acids in pupae and adults of the silkworm, Bombyx mori, was investigated. The rate of 14 CO 2 production from DL-cystine-1- 14 C varied during pupal development and also between male and female significantly; in the male the 14 CO 2 production increased markedly according to pupal development, whereas in the female it was kept at a low level until emergence, then increased slightly. Total sulfur in meconium of the female was less than a half the quantity of that of the male. However, in the ovary-ectomyzed female the amount of total sulfur in meconium and the rate of 14 CO 2 production were increased, almost similar to the case of the male. It was also confirmed that the 14 C of DL-cystine-1- 14 C was incorporated efficiently into egg protein at the end of pupal stage. These results indicated that the rate of degradation of sulfur amino acids in the pupal stage was much active in male than in female. Such a sexual difference is considered to be related closely to the formation of egg, especially egg-shell. (auth.)

  14. Phylogenetic analysis of Bombyx mori nucleopolyhedrovirus polyhedrin and p10 genes in wild isolates from Guangxi Zhuang Autonomous Region, China.

    Science.gov (United States)

    Liang, Xiang; Lu, Zhuan-Ling; Wei, Bing-Xing; Feng, Jian-Ling; Qu, Dacai; Luo, Ting Rong

    2013-02-01

    Bombyx mori nucleopolyhedrovirus (BmNPV) is a severe pathogen that seriously impacts the sericulture industry. In this study, 45 wild BmNPV isolates were collected from different silkworm-raising regions in China's Guangxi Zhuang Autonomous Region. Two highly expressed very late genes from each isolate, polyhedrin and p10, were sequenced and subjected to phylogenetic analysis. The polyhedrin gene was found to be highly conserved, while the p10 gene was more variable frequently harboring point mutations and displaying variations in codon use without obvious codon bias. The BmNPV isolates from Guangxi were separated into three main clades, I, II, and III, according to the p10 gene phylogenetic tree. The geographical distribution of clade I isolates in Guangxi showed a concentrated pattern and that of clade II isolates showed a connected pattern. Clade III isolates were irregularly scattered throughout Guangxi. Local transmission of this pathogen clearly occurred in the silkworm-raising regions in Guangxi. This study may provide some data on BmNPV transmission in the silkworm-raising regions and be helpful in devising strategies for the prevention and control of BmNPV disease.

  15. piggyBac transposon-derived targeting shRNA interference against the Bombyx mori nucleopolyhedrovirus (BmNPV).

    Science.gov (United States)

    Zhou, Fang; Chen, Rui-ting; Lu, Yan; Liang, Shuang; Wang, Mei-xian; Miao, Yun-gen

    2014-12-01

    The Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the most destructive diseases in silkworm, which has caused the main damage to sericulture industry. In this study, we developed a system of RNAi to prevent the BmNPV infection using the piggyBac transposon-derived targeting short hairpin RNA (shRNA) interference. The shRNAs targeting the genes of i.e.-1, lef-1, lef-2 and lef-3 of BmNPV were designed and used to inhibit the intracellular replication or multiplication of BmNPV in Bm cells. The highest activity was presented in the shRNA targeting the i.e.-1c of BmNPV, of which the inhibition rate reached 94.5 % in vitro. Further a stable Bm cell line of piggyBac transposon-derived targeting shRNA interference against BmNPV was established, which has a highly efficacious suppression on virus proliferation. These results indicated that the recombinant shRNA expression system was a useful tool for resistance to BmNPV in vitro. The approach by recombinant shRNAs opens a door of RNAi technology as a strategy that offering technically simpler, cheaper, and quicker gene knockdown for promising research and biotechnology application on silkworm lethal diseases.

  16. Genome-Wide Analysis of Differentially Expressed microRNA in Bombyx mori Infected with Nucleopolyhedrosis Virus.

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    Ping Wu

    Full Text Available Bombyx mori nucleopolyhedrosis virus (BmNPV is a major pathogen that threatens the growth and sustainability of the sericulture industry. Since microRNAs (miRNAs have been shown to play important roles in host-pathogen interactions, in this study we investigated the effects of BmNPV infection on silkworm microRNAs expression profile. To achieve this, we constructed and deep-sequenced two small RNA libraries generated from BmNPV infected and un-infected larvae. The results revealed that 38 silkworm miRNAs were differentially expressed after BmNPV infection. Based on the GO analysis, their predicted target genes were found to be involved in diverse functions such as binding, catalytic, virion and immune response to stimulus suggesting their potential roles in host-virus interactions. Using the dual-luciferase reporter assay, we confirmed that Bmo-miR-277-5p, up-regulated in BmNPV-infected larvae, targeted the B. mori DNA cytosine-5 methyltransferase (Dnmt2 gene which may play potential role in silkworm-BmNPV interaction. These results provide new insights into exploring the interaction mechanism between silkworm and BmNPV.

  17. Genome-Wide Analysis of Differentially Expressed microRNA in Bombyx mori Infected with Nucleopolyhedrosis Virus.

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    Wu, Ping; Jiang, Xiaoxu; Guo, Xijie; Li, Long; Chen, Tao

    2016-01-01

    Bombyx mori nucleopolyhedrosis virus (BmNPV) is a major pathogen that threatens the growth and sustainability of the sericulture industry. Since microRNAs (miRNAs) have been shown to play important roles in host-pathogen interactions, in this study we investigated the effects of BmNPV infection on silkworm microRNAs expression profile. To achieve this, we constructed and deep-sequenced two small RNA libraries generated from BmNPV infected and un-infected larvae. The results revealed that 38 silkworm miRNAs were differentially expressed after BmNPV infection. Based on the GO analysis, their predicted target genes were found to be involved in diverse functions such as binding, catalytic, virion and immune response to stimulus suggesting their potential roles in host-virus interactions. Using the dual-luciferase reporter assay, we confirmed that Bmo-miR-277-5p, up-regulated in BmNPV-infected larvae, targeted the B. mori DNA cytosine-5 methyltransferase (Dnmt2) gene which may play potential role in silkworm-BmNPV interaction. These results provide new insights into exploring the interaction mechanism between silkworm and BmNPV.

  18. Production Efficiency of Cocoon Shell of Silkworm, Bombyx mori L. (Bombycidae: Lepidoptera, as an Index for Evaluating the Nutritive Value of Mulberry, Morus sp. (Moraceae, Varieties

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    Jalaja Suresh Kumar

    2011-01-01

    Full Text Available The nutritional efficiency of mulberry leaves consumed by silkworms, Bombyx mori L., is usually evaluated in terms of the proportion of cocoon shell weight to the amount of food ingested. The production efficiency of cocoon shell is generally used to identify the superiority of a mulberry variety for silkworm rearing. In this study the production efficiency of cocoon shell was used as an index for evaluating the nutritive value of different mulberry varieties of India. Among the varieties, V-1, having highest production efficiency of cocoon shell with less amount of food ingested and highest digestibility, is regarded as the best suitable variety with nutritive values ideal for silkworm rearing.

  19. Identification of lipases involved in PBAN stimulated pheromone production in Bombyx mori using the DGE and RNAi approaches.

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    Mengfang Du

    Full Text Available BACKGROUND: Pheromone biosynthesis activating neuropeptide (PBAN is a neurohormone that regulates sex pheromone synthesis in female moths. Bombyx mori is a model organism that has been used to explore the signal transduction pattern of PBAN, which is mediated by a G-protein coupled receptor (GPCR. Although significant progress has been made in elucidating PBAN-regulated lipolysis that releases the precursor of the sex pheromone, little is known about the molecular components involved in this step. To better elucidate the molecular mechanisms of PBAN-stimulated lipolysis of cytoplasmic lipid droplets (LDs, the associated lipase genes involved in PBAN- regulated sex pheromone biosynthesis were identified using digital gene expression (DGE and subsequent RNA interference (RNAi. RESULTS: Three DGE libraries were constructed from pheromone glands (PGs at different developed stages, namely, 72 hours before eclosion (-72 h, new emergence (0 h and 72 h after eclosion (72 h, to investigate the gene expression profiles during PG development. The DGE evaluated over 5.6 million clean tags in each PG sample and revealed numerous genes that were differentially expressed at these stages. Most importantly, seven lipases were found to be richly expressed during the key stage of sex pheromone synthesis and release (new emergence. RNAi-mediated knockdown confirmed for the first time that four of these seven lipases play important roles in sex pheromone synthesis. CONCLUSION: This study has identified four lipases directly involved in PBAN-stimulated sex pheromone biosynthesis, which improve our understanding of the lipases involved in releasing bombykol precursors from triacylglycerols (TAGs within the cytoplasmic LDs.

  20. Genome-wide identification of polycomb target genes reveals a functional association of Pho with Scm in Bombyx mori.

    Science.gov (United States)

    Li, Zhiqing; Cheng, Daojun; Mon, Hiroaki; Tatsuke, Tsuneyuki; Zhu, Li; Xu, Jian; Lee, Jae Man; Xia, Qingyou; Kusakabe, Takahiro

    2012-01-01

    Polycomb group (PcG) proteins are evolutionarily conserved chromatin modifiers and act together in three multimeric complexes, Polycomb repressive complex 1 (PRC1), Polycomb repressive complex 2 (PRC2), and Pleiohomeotic repressive complex (PhoRC), to repress transcription of the target genes. Here, we identified Polycomb target genes in Bombyx mori with holocentric centromere using genome-wide expression screening based on the knockdown of BmSCE, BmESC, BmPHO, or BmSCM gene, which represent the distinct complexes. As a result, the expressions of 29 genes were up-regulated after knocking down 4 PcG genes. Particularly, there is a significant overlap between targets of BmPho (331 out of 524) and BmScm (331 out of 532), and among these, 190 genes function as regulator factors playing important roles in development. We also found that BmPho, as well as BmScm, can interact with other Polycomb components examined in this study. Further detailed analysis revealed that the C-terminus of BmPho containing zinc finger domain is involved in the interaction between BmPho and BmScm. Moreover, the zinc finger domain in BmPho contributes to its inhibitory function and ectopic overexpression of BmScm is able to promote transcriptional repression by Gal4-Pho fusions including BmScm-interacting domain. Loss of BmPho expression causes relocalization of BmScm into the cytoplasm. Collectively, we provide evidence of a functional link between BmPho and BmScm, and propose two Polycomb-related repression mechanisms requiring only BmPho associated with BmScm or a whole set of PcG complexes.

  1. Positional cloning of a Bombyx pink-eyed white egg locus reveals the major role of cardinal in ommochrome synthesis.

    Science.gov (United States)

    Osanai-Futahashi, M; Tatematsu, K-I; Futahashi, R; Narukawa, J; Takasu, Y; Kayukawa, T; Shinoda, T; Ishige, T; Yajima, S; Tamura, T; Yamamoto, K; Sezutsu, H

    2016-02-01

    Ommochromes are major insect pigments involved in coloration of compound eyes, eggs, epidermis and wings. In the silkworm Bombyx mori, adult compound eyes and eggs contain a mixture of the ommochrome pigments such as ommin and xanthommatin. Here, we identified the gene involved in ommochrome biosynthesis by positional cloning of B. mori egg and eye color mutant pink-eyed white egg (pe). The recessive homozygote of pe has bright red eyes and white or pale pink eggs instead of a normal dark coloration due to the decrease of dark ommochrome pigments. By genetic linkage analysis, we narrowed down the pe-linked region to ~258 kb, containing 17 predicted genes. RNA sequencing analyses showed that the expression of one candidate gene, the ortholog of Drosophila haem peroxidase cardinal, coincided with egg pigmentation timing, similar to other ommochrome-related genes such as Bm-scarlet and Bm-re. In two pe strains, a common missense mutation was found within a conserved motif of B. mori cardinal homolog (Bm-cardinal). RNA interference-mediated knockdown and transcription activator-like effector nuclease (TALEN)-mediated knockout of the Bm-cardinal gene produced the same phenotype as pe in terms of egg, adult eye and larval epidermis coloration. A complementation test of the pe mutant with the TALEN-mediated Bm-cardinal-deficient strain showed that the mutant phenotype could not be rescued, indicating that Bm-cardinal is responsible for pe. Moreover, knockdown of the cardinal homolog in Tribolium castaneum also induced red compound eyes. Our results indicate that cardinal plays a major role in ommochrome synthesis of holometabolous insects.

  2. Genetic analysis of the electrophysiological response to salicin, a bitter substance, in a polyphagous strain of the silkworm Bombyx mori.

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    Tetsuya Iizuka

    Full Text Available Sawa-J is a polyphagous silkworm (Bombyx mori L. strain that eats various plant leaves that normal silkworms do not. The feeding preference behavior of Sawa-J is controlled by one major recessive gene(s on the polyphagous (pph locus, and several minor genes; moreover, its deterrent cells possess low sensitivity to some bitter substances including salicin. To clarify whether taste sensitivity is controlled by the pph locus, we conducted a genetic analysis of the electrophysiological characteristics of the taste response using the polyphagous strain Sawa-J·lem, in which pph is linked to the visible larval marker lemon (lem on the third chromosome, and the normal strain Daiankyo, in which the wild-type gene of pph (+(pph is marked with Zebra (Ze. Maxillary taste neurons of the two strains had similar dose-response relationships for sucrose, inositol, and strychnine nitrate, but the deterrent cell of Sawa-J·lem showed a remarkably low sensitivity to salicin. The F(1 generation of the two strains had characteristics similar to the Daiankyo strain, consistent with the idea that pph is recessive. In the BF(1 progeny between F(1 females and Sawa-J·lem males where no crossing-over occurs, the lem and Ze phenotypes corresponded to different electrophysiological reactions to 25 mM salicin, indicating that the gene responsible for taste sensitivity to salicin is located on the same chromosome as the lem and Ze genes. The normal and weak reactions to 25 mM salicin were segregated in crossover-type larvae of the BF(1 progeny produced by a reciprocal cross, and the recombination frequency agreed well with the theoretical ratio for the loci of lem, pph, and Ze on the standard linkage map. These results indicate that taste sensitivity to salicin is controlled by the gene(s on the pph locus.

  3. Safety and immunogenicity of H5N1 influenza vaccine based on baculovirus surface display system of Bombyx mori.

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    Rongzhong Jin

    Full Text Available Avian influenza virus (H5N1 has caused serious infections in human beings. This virus has the potential to emerge as a pandemic threat in humans. Effective vaccines against H5N1 virus are needed. A recombinant Bombyx mori baculovirus, Bmg64HA, was constructed for the expression of HA protein of H5N1 influenza virus displaying on the viral envelope surface. The HA protein accounted for approximately 3% of the total viral proteins in silkworm pupae infected with the recombinant virus. Using a series of separation and purification methods, pure Bmgp64HA virus was isolated from these silkworm pupae bioreactors. Aluminum hydroxide adjuvant was used for an H5N1 influenza vaccine. Immunization with this vaccine at doses of 2 mg/kg and 0.67 mg/kg was carried out to induce the production of neutralizing antibodies, which protected monkeys against influenza virus infection. At these doses, the vaccine induced 1:40 antibody titers in 50% and 67% of the monkeys, respectively. The results of safety evaluation indicated that the vaccine did not cause any toxicity at the dosage as large as 3.2 mg/kg in cynomolgus monkeys and 1.6 mg/kg in mice. The results of dose safety evaluation of vaccine indicated that the safe dose of the vaccine were higher than 0.375 mg/kg in rats and 3.2 mg/kg in cynomolgus monkeys. Our work showed the vaccine may be a candidate for a highly effective, cheap, and safe influenza vaccine for use in humans.

  4. Bombyx mori histone methyltransferase BmAsh2 is essential for silkworm piRNA-mediated sex determination.

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    Li, Zhiqian; You, Lang; Yan, Dong; James, Anthony A; Huang, Yongping; Tan, Anjiang

    2018-02-01

    Sex determination is a hierarchically-regulated process with high diversity in different organisms including insects. The W chromosome-derived Fem piRNA has been identified as the primary sex determination factor in the lepidopteran insect, Bombyx mori, revealing a distinctive piRNA-mediated sex determination pathway. However, the comprehensive mechanism of silkworm sex determination is still poorly understood. We show here that the silkworm PIWI protein BmSiwi, but not BmAgo3, is essential for silkworm sex determination. CRISPR/Cas9-mediated depletion of BmSiwi results in developmental arrest in oogenesis and partial female sexual reversal, while BmAgo3 depletion only affects oogenesis. We identify three histone methyltransferases (HMTs) that are significantly down-regulated in BmSiwi mutant moths. Disruption one of these, BmAsh2, causes dysregulation of piRNAs and transposable elements (TEs), supporting a role for it in the piRNA signaling pathway. More importantly, we find that BmAsh2 mutagenesis results in oogenesis arrest and partial female-to-male sexual reversal as well as dysregulation of the sex determination genes, Bmdsx and BmMasc. Mutagenesis of other two HMTs, BmSETD2 and BmEggless, does not affect piRNA-mediated sex determination. Histological analysis and immunoprecipitation results support a functional interaction between the BmAsh2 and BmSiwi proteins. Our data provide the first evidence that the HMT, BmAsh2, plays key roles in silkworm piRNA-mediated sex determination.

  5. Identification of key uric acid synthesis pathway in a unique mutant silkworm Bombyx mori model of Parkinson's disease.

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    Hiroko Tabunoki

    Full Text Available Plasma uric acid (UA levels decrease following clinical progression and stage development of Parkinson's disease (PD. However, the molecular mechanisms underlying decreases in plasma UA levels remain unclear, and the potential to apply mutagenesis to a PD model has not previously been discovered. We identified a unique mutant of the silkworm Bombyx mori (B.mori op. Initially, we investigated the causality of the phenotypic "op" by microarray analysis using our constructed KAIKO functional annotation pipeline. Consequently, we found a novel UA synthesis-modulating pathway, from DJ-1 to xanthine oxidase, and established methods for large-scale analysis of gene expression in B. mori. We found that the mRNA levels of genes in this pathway were significantly lower in B. mori op mutants, indicating that downstream events in the signal transduction cascade might be prevented. Additionally, levels of B.mori tyrosine hydroxylase (TH and DJ-1 mRNA were significantly lower in the brain of B. mori op mutants. UA content was significantly lower in the B. mori op mutant tissues and hemolymph. The possibility that the B. mori op mutant might be due to loss of DJ-1 function was supported by the observed vulnerability to oxidative stress. These results suggest that UA synthesis, transport, elimination and accumulation are decreased by environmental oxidative stress in the B. mori op mutant. In the case of B. mori op mutants, the relatively low availability of UA appears to be due both to the oxidation of DJ-1 and to its expenditure to mitigate the effects of environmental oxidative stress. Our findings are expected to provide information needed to elucidate the molecular mechanism of decreased plasma UA levels in the clinical stage progression of PD.

  6. Isolation and bioactivities of a non-sericin component from cocoon shell silk sericin of the silkworm Bombyx mori.

    Science.gov (United States)

    Wang, Hai-Yan; Wang, Yuan-Jing; Zhou, Li-Xia; Zhu, Lin; Zhang, Yu-Qing

    2012-02-01

    The cocoon shell of the silkworm Bombyx mori consists of silk fibroin fiber (70%) surrounded by a sericin layer made up of sericin (25%) and non-sericin (5%) components. The non-sericin component which consists of carbohydrate, salt, wax, flavonoids and derivatives is often overlooked in applied research into sericin and its hydrolysate. Here, sericin and non-sericin compounds were obtained from the sericin layer of five types of cocoon shell by means of degumming in water followed by extraction and separation in ethanol. These ethanol extracts were found to mainly contain flavonoids and free amino acids possessing scavenging activities of the 2,2-diphenyl -1-picrylhydrazyl (DPPH) free radical and inhibiting activities of tyrosinase, which were much greater than the corresponding activities of the purified sericin proteins. The extracts also strongly inhibited α-glucosidase while the sericins had no such activity. In particular, the inhibitory activities of the ethanol extract of Daizo cocoons were much greater than those of the other cocoons. The IC(50) values of the Daizo cocoons for DPPH free radicals, tyrosinase, and α-glucosidase were 170, 27, and 110 μg mL(-1), respectively. The bioactivities of the non-sericin component were much higher than the activity of sericin alone. In addition, the in vivo test showed preliminarily that the administration of the non-sericin component had effectively resistant activity against streptozocin (STZ) oxidation and that of the purified sericin could also evidently decrease the induction ratio of diabetic mice induced by STZ. Therefore, ethanol extract protocols of the sericin layer of cocoon shells provide a novel stock which, together with sericin protein, has potential uses in functional food, biotechnological and medical applications.

  7. Fibroin and sericin from Bombyx mori silk stimulate cell migration through upregulation and phosphorylation of c-Jun.

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    Celia Martínez-Mora

    Full Text Available Wound healing is a biological process directed to the restoration of tissue that has suffered an injury. An important phase of wound healing is the generation of a basal epithelium able to wholly replace the epidermis of the wound. A broad range of products derived from fibroin and sericin from Bombyx mori silk are used to stimulate wound healing. However, so far the molecular mechanism underlying this phenomenon has not been elucidated. The aim of this work was to determine the molecular basis underlying wound healing properties of silk proteins using a cell model. For this purpose, we assayed fibroin and sericin in a wound healing scratch assay using MDA-MB-231 and Mv1Lu cells. Both proteins stimulated cell migration. Furthermore, treatment with sericin and fibroin involved key factors of the wound healing process such as upregulation of c-Jun and c-Jun protein phosphorylation. Moreover, fibroin and sericin stimulated the phosphorylation of ERK 1/2 and JNK 1/2 kinases. All these experiments were done in the presence of specific inhibitors for some of the cell signalling pathways referred above. The obtained results revealed that MEK, JNK and PI3K pathways are involved in fibroin and sericin stimulated cells migration. Inhibition of these three kinases prevented c-Jun upregulation and phosphorylation by fibroin or sericin. Fibroin and sericin were tested in the human keratinocyte cell line, HaCaT, with similar results. Altogether, our results showed that fibroin and sericin initiate cell migration by activating the MEK, JNK and PI3K signalling pathways ending in c-Jun activation.

  8. Tuning molecular weights of Bombyx mori (B. mori) silk sericin to modify its assembly structures and materials formation.

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    Yang, Mingying; Shuai, Yajun; Zhou, Guanshan; Mandal, Namita; Zhu, Liangjun; Mao, Chuanbin

    2014-08-27

    Bombyx mori (B. mori) silk sericin is a protein with features desirable as a biomaterial, such as increased hydrophilicity and biodegradation, as well as resistance to oxidation, bacteria, and ultraviolet light. In contrast to other widely studied B. mori silk proteins such as fibroin, sericin is still unexplored as a building block for fabricating biomaterial, and thus a facile technique of processing it into a material is needed. Here, electrospinning technology was used to fabricate it into biomaterials from two forms of B. mori silk sericin with different molecular weights, one is a low (12.0 kDa) molecular sericin (LS) form and another is a high (66.0 kDa) molecular weight sericin (HS) form. Circular dichroism (CD) spectra showed that LS in hexafluoroacetone (HFA) solvent adopted a predominantly random coil conformation, whereas HS tended to form a β-sheet structure along with a large content of random coils. In addition, LS and HS in HFA solvent were found to form cylinder-like smaller nanoparticles and larger irregular aggregates before electrospinning, respectively. As a result, biomaterials based on microparticles and nanofibers were successfully fabricated by electrospinning of LS and HS dissolved in HFA, respectively. The cell viability and differentiation assay indicated that nanofibers and microparticles improved cell adhesion, growth, and differentiation, proving that the scaffolds electrospun from sericin are biocompatible regardless of its molecular weight. The microparticles, not common in electrospinning of silk proteins reported previously, were found to promote the osteogenic differentiation of mesenchymal stem cells in comparison to the nanofibers. This study suggested that molecular weight of sericin mediates its secondary structure and assembly structure, which in turn leads to a control of final morphology of the electrospun materials. The microparticles and nanofibers of sericin can be potentially used as building blocks for fabricating

  9. Fibroin and sericin from Bombyx mori silk stimulate cell migration through upregulation and phosphorylation of c-Jun.

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    Martínez-Mora, Celia; Mrowiec, Anna; García-Vizcaíno, Eva María; Alcaraz, Antonia; Cenis, José Luis; Nicolás, Francisco José

    2012-01-01

    Wound healing is a biological process directed to the restoration of tissue that has suffered an injury. An important phase of wound healing is the generation of a basal epithelium able to wholly replace the epidermis of the wound. A broad range of products derived from fibroin and sericin from Bombyx mori silk are used to stimulate wound healing. However, so far the molecular mechanism underlying this phenomenon has not been elucidated. The aim of this work was to determine the molecular basis underlying wound healing properties of silk proteins using a cell model. For this purpose, we assayed fibroin and sericin in a wound healing scratch assay using MDA-MB-231 and Mv1Lu cells. Both proteins stimulated cell migration. Furthermore, treatment with sericin and fibroin involved key factors of the wound healing process such as upregulation of c-Jun and c-Jun protein phosphorylation. Moreover, fibroin and sericin stimulated the phosphorylation of ERK 1/2 and JNK 1/2 kinases. All these experiments were done in the presence of specific inhibitors for some of the cell signalling pathways referred above. The obtained results revealed that MEK, JNK and PI3K pathways are involved in fibroin and sericin stimulated cells migration. Inhibition of these three kinases prevented c-Jun upregulation and phosphorylation by fibroin or sericin. Fibroin and sericin were tested in the human keratinocyte cell line, HaCaT, with similar results. Altogether, our results showed that fibroin and sericin initiate cell migration by activating the MEK, JNK and PI3K signalling pathways ending in c-Jun activation.

  10. A genome-wide survey for host response of silkworm, Bombyx mori during pathogen Bacillus bombyseptieus infection.

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    Lulin Huang

    Full Text Available Host-pathogen interactions are complex relationships, and a central challenge is to reveal the interactions between pathogens and their hosts. Bacillus bombysepticus (Bb which can produces spores and parasporal crystals was firstly separated from the corpses of the infected silkworms (Bombyx mori. Bb naturally infects the silkworm can cause an acute fuliginosa septicaemia and kill the silkworm larvae generally within one day in the hot and humid season. Bb pathogen of the silkworm can be used for investigating the host responses after the infection. Gene expression profiling during four time-points of silkworm whole larvae after Bb infection was performed to gain insight into the mechanism of Bb-associated host whole body effect. Genome-wide survey of the host genes demonstrated many genes and pathways modulated after the infection. GO analysis of the induced genes indicated that their functions could be divided into 14 categories. KEGG pathway analysis identified that six types of basal metabolic pathway were regulated, including genetic information processing and transcription, carbohydrate metabolism, amino acid and nitrogen metabolism, nucleotide metabolism, metabolism of cofactors and vitamins, and xenobiotic biodegradation and metabolism. Similar to Bacillus thuringiensis (Bt, Bb can also induce a silkworm poisoning-related response. In this process, genes encoding midgut peritrophic membrane proteins, aminopeptidase N receptors and sodium/calcium exchange protein showed modulation. For the first time, we found that Bb induced a lot of genes involved in juvenile hormone synthesis and metabolism pathway upregulated. Bb also triggered the host immune responses, including cellular immune response and serine protease cascade melanization response. Real time PCR analysis showed that Bb can induce the silkworm systemic immune response, mainly by the Toll pathway. Anti-microorganism peptides (AMPs, including of Attacin, Lebocin, Enbocin, Gloverin

  11. Transcriptome analysis of integument differentially expressed genes in the pigment mutant (quail during molting of silkworm, Bombyx mori.

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    Hongyi Nie

    Full Text Available In the silkworm Bombyx mori, pigment mutants with diverse body colors have been maintained throughout domestication for about 5000 years. The silkworm larval body color is formed through the mutual interaction of melanin, ommochromes, pteridines and uric acid. These pigments/compounds are synthesized by the cooperative action of various genes and enzymes. Previous reports showed that melanin, ommochrome and pteridine are increased in silkworm quail (q mutants. To understand the pigment increase and alterations in pigment synthesis in q mutant, transcriptome profiles of the silkworm integument were investigated at 16 h after head capsule slippage in the fourth molt in q mutants and wild-type (Dazao. Compared to the wild-type, 1161 genes were differentially expressed in the q mutant. Of these modulated genes, 62.4% (725 genes were upregulated and 37.6% (436 genes were downregulated in the q mutant. The molecular function of differently expressed genes was analyzed by Blast2GO. The results showed that upregulated genes were mainly involved in protein binding, small molecule binding, transferase activity, nucleic acid binding, specific DNA-binding transcription factor activity and chromatin binding, while exclusively down-expressed genes functioned in oxidoreductase activity, cofactor binding, tetrapyrrole binding, peroxidase activity and pigment binding. We focused on genes related to melanin, pteridine and ommochrome biosynthesis; transport of uric acid; and juvenile hormone metabolism because of their importance in integument coloration during molting. This study identified differently expressed genes implicated in silkworm integument formation and pigmentation using silkworm q mutant. The results estimated the number and types of genes that drive new integument formation.

  12. The specificity of immune priming in silkworm, Bombyx mori, is mediated by the phagocytic ability of granular cells.

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    Wu, Gongqing; Li, Mei; Liu, Yi; Ding, Ying; Yi, Yunhong

    2015-10-01

    In the past decade, the phenomenon of immune priming was documented in many invertebrates in a large number of studies; however, in most of these studies, behavioral evidence was used to identify the immune priming. The underlying mechanism and the degree of specificity of the priming response remain unclear. We studied the mechanism of immune priming in the larvae of the silkworm, Bombyx mori, and analyzed the specificity of the priming response using two closely related Gram-negative pathogenic bacteria (Photorhabdus luminescens TT01 and P. luminescens H06) and one Gram-positive pathogenic bacterium (Bacillus thuringiensis HD-1). Primed with heat-killed bacteria, the B. mori larvae were more likely to survive subsequent homologous exposure (the identical bacteria used in the priming and in the subsequent challenge) than heterologous (different bacteria used in the priming and subsequent exposure) exposure to live bacteria. This result indicated that the B. mori larvae possessed a strong immune priming response and revealed a degree of specificity to TT01, H06 and HD-1 bacteria. The degree of enhanced immune protection was positively correlated with the level of phagocytic ability of the granular cells and the antibacterial activity of the cell-free hemolymph. Moreover, the granular cells of the immune-primed larvae increased the phagocytosis of a previously encountered bacterial strain compared with other bacteria. Thus, the enhanced immune protection of the B. mori larvae after priming was mediated by the phagocytic ability of the granular cells and the antibacterial activity of the hemolymph; the specificity of the priming response was primarily attributed to the phagocytosis of bacteria by the granular cells. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Co-sensitization to silkworm moth (Bombyx mori and 9 inhalant allergens among allergic patients in Guangzhou, Southern China.

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    Baoqing Sun

    Full Text Available OBJECTIVES: This study aimed to investigate the profile of sensitization to silkworm moth (Bombyx mori and other 9 common inhalant allergens among patients with allergic diseases in southern China. METHODS: A total of 175 patients were tested for serum sIgE against silkworm moth in addition to combinations of other allergens: Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicalis, Blattella germanica, Periplaneta americana, cat dander, dog dander, Aspergillus fumigatus and Artemisia vulgaris by using the ImmunoCAP system. Correlation between sensitization to silkworm moth and to the other allergens was analyzed. RESULTS: Of the 175 serum samples tested, 86 (49.14% were positive for silkworm moth sIgE. With high concordance rates, these silkworm moth sensitized patients were concomitantly sensitized to Dermatophagoides pteronyssinus (94.34%, Dermatophagoides farinae (86.57%, Blomia tropicalis (93.33%, Blattella germanica (96.08%, and Periplaneta americana (79.41%. Moreover, there was a correlation in serum sIgE level between silkworm moth and Dermatophagoides pteronyssinus (r = 0.518, Dermatophagoides farinae (r = 0.702, Blomia tropicalis (r = 0.701, Blattella germanica (r = 0.878, and Periplaneta americana (r = 0.531 among patients co-sensitized to silkworm moth and each of these five allergens. CONCLUSION: In southern Chinese patients with allergic diseases, we showed a high prevalence of sensitization to silkworm moth, and a co-sensitization between silkworm moth and other five common inhalant allergens. Further serum inhibition studies are warranted to verify whether cross-reactivity exists among these allergens.

  14. Bombyx mori P-element Somatic Inhibitor (BmPSI Is a Key Auxiliary Factor for Silkworm Male Sex Determination.

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    Jun Xu

    2017-01-01

    Full Text Available Manipulation of sex determination pathways in insects provides the basis for a wide spectrum of strategies to benefit agriculture and public health. Furthermore, insects display a remarkable diversity in the genetic pathways that lead to sex differentiation. The silkworm, Bombyx mori, has been cultivated by humans as a beneficial insect for over two millennia, and more recently as a model system for studying lepidopteran genetics and development. Previous studies have identified the B. mori Fem piRNA as the primary female determining factor and BmMasc as its downstream target, while the genetic scenario for male sex determination was still unclear. In the current study, we exploite the transgenic CRISPR/Cas9 system to generate a comprehensive set of knockout mutations in genes BmSxl, Bmtra2, BmImp, BmImpM, BmPSI and BmMasc, to investigate their roles in silkworm sex determination. Absence of Bmtra2 results in the complete depletion of Bmdsx transcripts, which is the conserved downstream factor in the sex determination pathway, and induces embryonic lethality. Loss of BmImp or BmImpM function does not affect the sexual differentiation. Mutations in BmPSI and BmMasc genes affect the splicing of Bmdsx and the female reproductive apparatus appeared in the male external genital. Intriguingly, we identify that BmPSI regulates expression of BmMasc, BmImpM and Bmdsx, supporting the conclusion that it acts as a key auxiliary factor in silkworm male sex determination.

  15. A genome-wide survey for host response of silkworm, Bombyx mori during pathogen Bacillus bombyseptieus infection.

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    Huang, Lulin; Cheng, Tingcai; Xu, Pingzhen; Cheng, Daojun; Fang, Ting; Xia, Qingyou

    2009-12-01

    Host-pathogen interactions are complex relationships, and a central challenge is to reveal the interactions between pathogens and their hosts. Bacillus bombysepticus (Bb) which can produces spores and parasporal crystals was firstly separated from the corpses of the infected silkworms (Bombyx mori). Bb naturally infects the silkworm can cause an acute fuliginosa septicaemia and kill the silkworm larvae generally within one day in the hot and humid season. Bb pathogen of the silkworm can be used for investigating the host responses after the infection. Gene expression profiling during four time-points of silkworm whole larvae after Bb infection was performed to gain insight into the mechanism of Bb-associated host whole body effect. Genome-wide survey of the host genes demonstrated many genes and pathways modulated after the infection. GO analysis of the induced genes indicated that their functions could be divided into 14 categories. KEGG pathway analysis identified that six types of basal metabolic pathway were regulated, including genetic information processing and transcription, carbohydrate metabolism, amino acid and nitrogen metabolism, nucleotide metabolism, metabolism of cofactors and vitamins, and xenobiotic biodegradation and metabolism. Similar to Bacillus thuringiensis (Bt), Bb can also induce a silkworm poisoning-related response. In this process, genes encoding midgut peritrophic membrane proteins, aminopeptidase N receptors and sodium/calcium exchange protein showed modulation. For the first time, we found that Bb induced a lot of genes involved in juvenile hormone synthesis and metabolism pathway upregulated. Bb also triggered the host immune responses, including cellular immune response and serine protease cascade melanization response. Real time PCR analysis showed that Bb can induce the silkworm systemic immune response, mainly by the Toll pathway. Anti-microorganism peptides (AMPs), including of Attacin, Lebocin, Enbocin, Gloverin and Moricin

  16. A single sex pheromone receptor determines chemical response specificity of sexual behavior in the silkmoth Bombyx mori.

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    Takeshi Sakurai

    2011-06-01

    Full Text Available In insects and other animals, intraspecific communication between individuals of the opposite sex is mediated in part by chemical signals called sex pheromones. In most moth species, male moths rely heavily on species-specific sex pheromones emitted by female moths to identify and orient towards an appropriate mating partner among a large number of sympatric insect species. The silkmoth, Bombyx mori, utilizes the simplest possible pheromone system, in which a single pheromone component, (E, Z-10,12-hexadecadienol (bombykol, is sufficient to elicit full sexual behavior. We have previously shown that the sex pheromone receptor BmOR1 mediates specific detection of bombykol in the antennae of male silkmoths. However, it is unclear whether the sex pheromone receptor is the minimally sufficient determination factor that triggers initiation of orientation behavior towards a potential mate. Using transgenic silkmoths expressing the sex pheromone receptor PxOR1 of the diamondback moth Plutella xylostella in BmOR1-expressing neurons, we show that the selectivity of the sex pheromone receptor determines the chemical response specificity of sexual behavior in the silkmoth. Bombykol receptor neurons expressing PxOR1 responded to its specific ligand, (Z-11-hexadecenal (Z11-16:Ald, in a dose-dependent manner. Male moths expressing PxOR1 exhibited typical pheromone orientation behavior and copulation attempts in response to Z11-16:Ald and to females of P. xylostella. Transformation of the bombykol receptor neurons had no effect on their projections in the antennal lobe. These results indicate that activation of bombykol receptor neurons alone is sufficient to trigger full sexual behavior. Thus, a single gene defines behavioral selectivity in sex pheromone communication in the silkmoth. Our findings show that a single molecular determinant can not only function as a modulator of behavior but also as an all-or-nothing initiator of a complex species

  17. Phoxim-induced damages of Bombyx mori larval midgut and titanium dioxide nanoparticles protective role under phoxim-induced toxicity.

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    Su, Junju; Li, Bing; Cheng, Shen; Zhu, Zhou; Sang, Xuezi; Gui, Suxin; Xie, Yi; Sun, Qingqing; Cheng, Zhe; Cheng, Jie; Hu, Rengping; Shen, Weide; Xia, Qingyou; Zhao, Ping; Hong, Fashui

    2014-12-01

    Phoxim (O,O-diethyl O-(alpha-cyanobenzylideneamino) phosphorothioate) is a powerful organophosphorus pesticide with high potential for Bombyx mori larvae of silkworm exposure. However, it is possible that during the phoxim metabolism, there is generation of reactive oxygen species (ROS) and phoxim may produce oxidative stress and neurotoxicity in an intoxicated silkworm. Titanium dioxide nanoparticles (TiO2 NPs) pretreatment has been demonstrated to increase antioxidant capacity and acetylcholinesterase (AChE) activity in organisms. This study was, therefore, undertaken to determine phoxim-induced oxidative stress and neurotoxicity to determine whether phoxim intoxication alters the antioxidant system and AChE activity in the B. mori larval midgut, and to determine whether TiO2 NPs pretreatment attenuates phoxim-induced toxicity. The findings suggested that phoxim exposure decreased survival of B. mori larvae, increased malondialdehyde (MDA), carbonyl and 8-OHdG levels, and ROS accumulation in the midgut. Furthermore, phoxim significantly decreased the activities of AChE, superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), glutathione-S-transferase (GST), and levels of ascorbic acid (AsA), reduced glutathione (GSH), and thiol in the midgut. TiO2 pretreatment, however, could increase AChE activity, and remove ROS via activating SOD, CAT, APX, GR, and GST, and accelerating AsA-GSH cycle, thus attenuated lipid, protein, and DNA peroxidation and improve B. mori larval survival under phoxim-induced toxicity. Moreover, this experimental system would help nanomaterials to be applied in the sericulture. © 2013 Wiley Periodicals, Inc.

  18. Effects of the biosynthesis and signaling pathway of ecdysterone on silkworm (Bombyx mori) following exposure to titanium dioxide nanoparticles.

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    Li, Fanchi; Gu, Zhiya; Wang, Binbin; Xie, Yi; Ma, Lie; Xu, Kaizun; Ni, Min; Zhang, Hua; Shen, Weide; Li, Bing

    2014-08-01

    Silkworm (Bombyx mori), a model Lepidoptera insect, is economically important. Its growth and development are regulated by endogenous hormones. During the process of transition from larvae to pupae, 20-hydroxyecdysone (20E) plays an important role. The recent surge in consumer products and applications using metallic nanoparticles has increased the possibility of human or ecosystem exposure due to their unintentional release into the environment. We investigated the effects of exposure to titanium dioxide nanoparticles (TiO2 NPs) on the action of 20E in B. mori. Titanium dioxide nanoparticle treatment shortened the molting duration by 8 hr and prolonged the molting peak period by 10 %. Solexa sequencing profiled the changes in gene expression in the brain of fifth-instar B. mori in response to TiO2NPS exposure for 72 hr, to address the effects on hormone metabolism and regulation. Thirty one genes were differentially expressed. The transcriptional levels of pi3k and P70S6K, which are involved in the target of the rapamycin (TOR) signaling pathway, were up-regulated. Transcriptional levels of four cytochrome P450 genes, which are involved in 20E biosynthesis, at different developmental stages (48, 96, 144, and 192 hr) at 5th instars of all displayed trends of increasing expression. Simultaneously, the ecdysterone receptors, also displayed increasing trends. The 20E titers at four developmental stages during the 5th instar were 1.26, 1.23, 1.72, and 2.16 fold higher, respectively, than the control group. These results indicate that feeding B. mori with TiO2 NPs stimulates 20E biosynthesis, shortens the developmental progression, and reduces the duration of molting. Thus, application of TiO2 NPs is of high significance for saving the labor force in sericulture, and our research provides a reference for the ecological problems in the field of Lepidoptera exposured to titanium dioxide nanoparticles.

  19. Oxidative stress and cytotoxicity elicited lipid peroxidation in hemocytes of Bombyx mori larva infested with dipteran parasitoid, Exorista bombycis.

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    Pooja, Makwana; Pradeep, Appukuttan Nair R; Hungund, Shambhavi P; Sagar, Chandrashekhar; Ponnuvel, Kangayam M; Awasthi, Arvind K; Trivedy, Kanika

    2017-12-20

    Parasitization of silkworm, Bombyx mori by invasive larva of dipteran parasitoid Exorista bombycis caused upto 20% revenue loss in sericulture. The parasitism was successful by suppressing host immune system however mechanism of immune suppression induced by E. bombycis is unknown which is unravelled here. The infestation induced cytotoxic symptoms in host hemocytes, such as vacuolated cytoplasm, porous plasma membrane, indented nuclei with condensed chromatin and dilated RER. One of the markers of necrosis is cell permeabilization, which can be measured as released lactate dehydrogenase (LDH). LDH level showed significantly (P<0.01) high release into extracellular medium in vitro after exposure of hemocytes to parasitoid larval tissue protein compared with control revealing membrane permeability and loss of cell integrity. At five minutes after exposure, cytotoxicity was 43% and was increased to 99% at 3h. The cytotoxicity is signalled by increased content of hydrogen peroxide (H2O2) causing lipid peroxidation followed by porosity in plasma membrane. A test for lipid peroxidation by measurement of lipid peroxidation breakdown product, malondialdehyde (MDA) revealed significant increase in peroxidation from one to 24 h post-invasion, with maximum at 12 h (P<0.008). Level of reactive oxygen species measured as H2O2 production increased from 6 to 12 h post-invasion and continued to increase significantly (P<0.03) reaching maximum at 48 h. These observations reveal that dipteran endoparasitoid invasion induced H2O2 production in the hemocytes causing cytotoxicity, lipid peroxidation and membrane porosity that suppressed both humoral- and cell-mediated immune responses of hemocytes in B. mori.

  20. Heterosis manifestations by survival and larval duration of F Bombyx mori L. Hybrids reared 1 with artificial diet

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    R. Guncheva

    2016-12-01

    Full Text Available Abstract. The aim of this paper was to study the influence of the degree of heterosis (compared to higher parent value- HP and mean parental values- MP on the signs, survival and duration of development in F hybrids of Bombyx mori L. from different geographical origin, fed by artificial diet. The study was conducted 1 at the Training Experimental Station of the Sericulture section of the Faculty of Agriculture at Trakia University. A total of 10 hybrids were tested featuring 6 breeds from the genetic bank of Sericulture and Agriculture Experiment Station (SAES-Vratsa. Silkworms were reared with artificial diet containing 15% powder of dried mulberry leaf produced at SAES-Vratsa and prepared by methods, developed by the manufacturer, whereby 250g of dry substance and 675ml of distilled water are homogenized using a mixer. The mixture is cured thermally in a microwave for 10min. at ~800W. The receptivity of specimens was -st determined based on the results from the survival and the duration in the I age of the larval stage. In heterogeneous crossbreds higher survival rate in the first age is exhibited by hybrids in which a Japanese type breed was used as dam. Homogeneous crossbreds created with the participation of Japanese breeds excel in both analyzed traits those involving a Chinese type. For the survival trait heterosis manifestations to higher parent value (HP and mean parental values (MP, were detected in the majority of hybrids (90.91%. Regarding the duration of development, heterosis effect to MP was observed in all F hybrids, 1 while to HP heterosis is exhibited by 60% of the tested F hybrids.

  1. The silkworm Bombyx mori cuticular protein CPR55 gene is regulated by the transcription factor βFTZ-F1

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    Md. Saheb Ali

    2016-01-01

    Full Text Available The insect cuticle is composed of various proteins and formed during the moult under a complex biological process that depends on the cross talk between hormone levels and gene expression. In the present study, we aimed to clarify the ecdysone-dependent temporal regulation mechanisms of cuticular proteins expression and the underlying control of Bombyx mori metamorphosis. The expression of CPR55 was observed from the W3 early stage and peaked at pupation when the ecdysteroid titre declined. CPR55 was induced by the ecdysone pulse, and their expression peaked at 24 h after transfer to a hormone free medium. Transcripts of CPR55 were neither observed after the 20E pulse treatment in the presence of cycloheximide nor after the addition of 20E in V4 wing discs. We analysed the upstream region of the CPR55 gene using a transient reporter assay with a gene gun system which identified only one βFTZ-F1 binding site important for cis-acting elements for the transcription activation of the luciferase reporter gene by an ecdysone pulse. Site-directed mutagenesis of this element in the context of the 589-bp promoter fragment drastically decreased the reporter activity. The nuclear protein bound to βFTZ-F1 sites was identified by an electrophoretic mobility shift assay suggesting that CPR55 expression was regulated by βFTZ-F1 through the ecdysone pulse. The results confirmed that transcription factor, BmβFTZ-F1, binds to the cis-regulatory elements in the promoter of the gene coding for cuticle protein, CPR55, and regulates its expression during B. mori metamorphosis.

  2. Changes in the activity and the expression of detoxification enzymes in silkworms (Bombyx mori) after phoxim feeding.

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    Wang, Y H; Gu, Z Y; Wang, J M; Sun, S S; Wang, B B; Jin, Y Q; Shen, W D; Li, B

    2013-01-01

    Silkworm (Bombyx mori) is an economically important insect. However, non-cocoon caused by chemical insecticide poisoning has largely hindered the development of sericulture. To explore the roles of detoxification enzymes in B. mori after insecticide poisoning, we monitored the activity changes of cytochrome P450 monooxygenase, glutathione-S-transferase, and carboxylesterase in B. mori midgut and fatbody after phoxim feeding. At the same time, the expression levels of detoxification enzyme-related genes were also determined by real-time quantitative PCR. Compare to the control levels, the activity of P450 in the midgut and fatbody was increased to 1.72 and 6.72 folds; the activity of GST was no change in midgut, and in fatbody increased to 1.11 folds; the activity of carboxylesterase in the midgut was decreased to 0.69 folds, and in fatbody increased to 1.13 folds. Correspondingly, the expression levels of detoxifying enzyme genes CYP6ae22, CYP9a21, GSTo1 and Bmcce were increased to 15.99, 3.32, 1.86 and 2.30 folds in the midgut and to 3.58, 1.84, 2.14 and 4.21 folds in the fatbody after phoxim treatment. These results demonstrated the important roles of detoxification enzymes in phoxim metabolism. In addition, the detected activities of such enzymes were generally lower than those in cotton bollworms (Helicoverpa armigera), which may contribute to the high susceptibility of B. mori to insecticides. Our findings laid the foundation for further investigations of the molecular mechanisms of organophosphorus pesticide metabolism in B. mori. Copyright © 2012 Elsevier Inc. All rights reserved.

  3. The FOXO transcription factor controls insect growth and development by regulating juvenile hormone degradation in the silkworm, Bombyx mori.

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    Zeng, Baosheng; Huang, Yuping; Xu, Jun; Shiotsuki, Takahiro; Bai, Hua; Palli, Subba Reddy; Huang, Yongping; Tan, Anjiang

    2017-07-14

    Forkhead box O (FOXO) functions as the terminal transcription factor of the insulin signaling pathway and regulates multiple physiological processes in many organisms, including lifespan in insects. However, how FOXO interacts with hormone signaling to modulate insect growth and development is largely unknown. Here, using the transgene-based CRISPR/Cas9 system, we generated and characterized mutants of the silkworm Bombyx mori FOXO ( BmFOXO ) to elucidate its physiological functions during development of this lepidopteran insect. The BmFOXO mutant (FOXO-M) exhibited growth delays from the first larval stage and showed precocious metamorphosis, pupating at the end of the fourth instar (trimolter) rather than at the end of the fifth instar as in the wild-type (WT) animals. However, different from previous reports on precocious metamorphosis caused by juvenile hormone (JH) deficiency in silkworm mutants, the total developmental time of the larval period in the FOXO-M was comparable with that of the WT. Exogenous application of 20-hydroxyecdysone (20E) or of the JH analog rescued the trimolter phenotype. RNA-seq and gene expression analyses indicated that genes involved in JH degradation but not in JH biosynthesis were up-regulated in the FOXO-M compared with the WT animals. Moreover, we identified several FOXO-binding sites in the promoter of genes coding for JH-degradation enzymes. These results suggest that FOXO regulates JH degradation rather than its biosynthesis, which further modulates hormone homeostasis to control growth and development in B. mori In conclusion, we have uncovered a pivotal role for FOXO in regulating JH signaling to control insect development. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. Genome-wide identification of polycomb target genes reveals a functional association of Pho with Scm in Bombyx mori.

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    Zhiqing Li

    Full Text Available Polycomb group (PcG proteins are evolutionarily conserved chromatin modifiers and act together in three multimeric complexes, Polycomb repressive complex 1 (PRC1, Polycomb repressive complex 2 (PRC2, and Pleiohomeotic repressive complex (PhoRC, to repress transcription of the target genes. Here, we identified Polycomb target genes in Bombyx mori with holocentric centromere using genome-wide expression screening based on the knockdown of BmSCE, BmESC, BmPHO, or BmSCM gene, which represent the distinct complexes. As a result, the expressions of 29 genes were up-regulated after knocking down 4 PcG genes. Particularly, there is a significant overlap between targets of BmPho (331 out of 524 and BmScm (331 out of 532, and among these, 190 genes function as regulator factors playing important roles in development. We also found that BmPho, as well as BmScm, can interact with other Polycomb components examined in this study. Further detailed analysis revealed that the C-terminus of BmPho containing zinc finger domain is involved in the interaction between BmPho and BmScm. Moreover, the zinc finger domain in BmPho contributes to its inhibitory function and ectopic overexpression of BmScm is able to promote transcriptional repression by Gal4-Pho fusions including BmScm-interacting domain. Loss of BmPho expression causes relocalization of BmScm into the cytoplasm. Collectively, we provide evidence of a functional link between BmPho and BmScm, and propose two Polycomb-related repression mechanisms requiring only BmPho associated with BmScm or a whole set of PcG complexes.

  5. Transgenic Expression of the piRNA-Resistant Masculinizer Gene Induces Female-Specific Lethality and Partial Female-to-Male Sex Reversal in the Silkworm, Bombyx mori.

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    Hiroki Sakai

    2016-08-01

    Full Text Available In Bombyx mori (B. mori, Fem piRNA originates from the W chromosome and is responsible for femaleness. The Fem piRNA-PIWI complex targets and cleaves mRNAs transcribed from the Masc gene. Masc encodes a novel CCCH type zinc-finger protein and is required for male-specific splicing of B. mori doublesex (Bmdsx transcripts. In the present study, several silkworm strains carrying a transgene, which encodes a Fem piRNA-resistant Masc mRNA (Masc-R, were generated. Forced expression of the Masc-R transgene caused female-specific lethality during the larval stages. One of the Masc-R strains weakly expressed Masc-R in various tissues. Females heterozygous for the transgene expressed male-specific isoform of the Bombyx homolog of insulin-like growth factor II mRNA-binding protein (ImpM and Bmdsx. All examined females showed a lower inducibility of vitellogenin synthesis and exhibited abnormalities in the ovaries. Testis-like tissues were observed in abnormal ovaries and, notably, the tissues contained considerable numbers of sperm bundles. Homozygous expression of the transgene resulted in formation of the male-specific abdominal segment in adult females and caused partial male differentiation in female genitalia. These results strongly suggest that Masc is an important regulatory gene of maleness in B. mori.

  6. Characterization and expression analysis of peroxiredoxin family genes from the silkworm Bombyx mori in response to phoxim and chlorpyrifos.

    Science.gov (United States)

    Shi, Gui-Qin; Zhang, Ze; Jia, Kun-Lun; Zhang, Kun; An, Dong-Xu; Wang, Gang; Zhang, Bao-Long; Yin, He-Nan

    2014-09-01

    The organophosphorus pesticide poisoning of the silkworm Bombyx mori is one of the major events causing serious damage to sericulture. Some antioxidant enzymes play roles in regulating generation of reactive oxygen species (ROS) by pesticides including phoxim and chlorpyrifos, but relatively little is known about their effects on the silkworm peroxiredoxin family genes. Here, five peroxiredoxin (Prx) genes have been identified in silkworm genome, and Prx genes of silkworm and mammalian homologs have apparent ortholog relationship. Based on the genomic DNA sequence, putative 5'-flanking region of five BmPrxs were obtained and the transcription factor binding sites were predicted. Their expression profiles exposed to different concentrations of phoxim and chlorpyrifos for 24 h, 48 h and 72 h in midgut of silkworm were investigated using quantitative RT-PCR (qRT-PCR). The results showed that five BmPrxs and dual oxidase (BmDUOX) gene were all expressed in midgut of silkworm. After feeding with 0.375 mg/L and 0.75 mg/L phoxim, the transcription levels of BmPrx3 and BmPrx5 that can be located in mitochondria reached their peak levels at an early time point (24h). However, the transcription levels of BmPrx4 and BmPrx6 that can be addressed to secrete from the cell and cytosol, respectively, reached their peak levels at a later time point (72 h). Similar to expose to phoxim, the transcription levels of BmPrx3 and BmPrx5 that can be located in mitochondria reached their peak levels at an early time point (24 h) under chlorpyrifos stress. However, the transcription levels of BmPrx4 and BmPrx6 that can be addressed to secrete from the cell and cytosol, respectively, reached their peak levels at a later time point (72 h) under chlorpyrifos stress. These results revealed that BmPrxs that can be located in mitochondria were able to protect cells even more efficiently than cytosolic from an oxidative stress caused by OP. In addition, BmDUOX was also induced by phomix and

  7. Microstructural Parameters in 8 MeV Electron-Irradiated BOMBYX MORI Silk Fibers by Wide-ANGLE X-Ray Scattering Studies (waxs)

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    Sangappa, Asha, S.; Sanjeev, Ganesh; Subramanya, G.; Parameswara, P.; Somashekar, R.

    2010-01-01

    The present work looks into the microstructural modification in electron irradiated Bombyx mori P31 silk fibers. The irradiation process was performed in air at room temperature using 8 MeV electron accelerator at different doses: 0, 25, 50 and 100 kGy. Irradiation of polymer is used to cross-link or degrade the desired component or to fix the polymer morphology. The changes in microstructural parameters in these natural polymer fibers have been computed using wide angle X-ray scattering (WAXS) data and employing line profile analysis (LPA) using Fourier transform technique of Warren. Exponential, Lognormal and Reinhold functions for the column length distributions have been used for the determination of crystal size, lattice strain and enthalpy parameters.

  8. MICROSTRUCTURAL PARAMETERS IN 8 MeV ELECTRON‐IRRADIATED BOMBYX MORI SILK FIBERS BY Wide‐ANGLE X‐RAY SCATTERING STUDIES (WAXS)

    International Nuclear Information System (INIS)

    Sangappa,; Asha, S; Sanjeev, Ganesh; Subramanya, G; Parameswara, P; Somashekar, R

    2010-01-01

    The present work looks into the microstructural modification in electron irradiated Bombyx mori P31 silk fibers. The irradiation process was performed in air at room temperature using 8 MeV electron accelerator at different doses: 0, 25, 50 and 100 kGy. Irradiation of polymer is used to cross‐link or degrade the desired component or to fix the polymer morphology. The changes in microstructural parameters in these natural polymer fibers have been computed using wide angle X‐ray scattering (WAXS) data and employing line profile analysis (LPA) using Fourier transform technique of Warren. Exponential, Lognormal and Reinhold functions for the column length distributions have been used for the determination of crystal size, lattice strain and enthalpy parameters.

  9. cDNA cloning and expression of Bacillus thuringiensis Cry1Aa toxin binding 120 kDa aminopeptidase N from Bombyx mori.

    Science.gov (United States)

    Yaoi, K; Nakanishi, K; Kadotani, T; Imamura, M; Koizumi, N; Iwahana, H; Sato, R

    1999-01-18

    Bacillus thuringiensis Cry1Aa toxin binds to a 120 kDa putative receptor protein in the Bombyx mori midgut. Recently, this protein was purified and identified as glycosyl-phosphatidylinositol (GPI) anchored aminopeptidase N (APN). In this study, a full-length cDNA thought to encode this 120 kDa APN was isolated and sequenced. It has a 2958 bp ORF encoding 986 amino acids. In the deduced amino acid sequence, we identified GPI-anchor and zinc-metallopeptidase signals, which are the same as those of APNs of other insects that are reported to be putative Cry1 toxin receptors. The B. mori APN amino acid sequence also has a high similarity with those of the other APNs. Subsequently, the recombinant APN was expressed by Escherichia coli and its Cry1Aa toxin binding ability was analyzed. Ligand blotting showed that Cry1Aa toxin bound to the recombinant APN.

  10. Cloning of anti-lPS factor cDNA from Tachypleus tridentatus, expression in Bombyx mori larvae and its biological activity in vitro.

    Science.gov (United States)

    Wang, Dong-Ning; Liu, Jie-Wu; Yang, Guan-Zhen; Zhang, Wei-Jie; Wu, Xiang-Fu

    2002-05-01

    In this article we report the cloning and expression of a cDNA encoding Tachypleus anti-lipopolysaccharide (LPS) factor, which is of interest for use as a potential inhibitor of the common core subunit of Gram-negative bacterial endotoxins. First, two degenerate primers were designed based on the sequence homology of anti-LPS factors purified from different species of horseshoe crab. The total RNA was extracted from amebocytes of Tachypleus tridentatus. The cDNA was then obtained by using the RT-PCR methods. Second, the cDNA of Tachypleus anti-LPS factor (TALF) was expressed in Bombyx mori larvae using baculovirus expression system, which showed a yield of up to 600 mg/L. Last, we determined the biological activity of the recombinant proteins by LPS neutralization assay and bacteriostatic assay in vitro.

  11. MICROSTRUCTURAL PARAMETERS IN 8 MeV ELECTRON‐IRRADIATED BOMBYX MORI SILK FIBERS BY Wide‐ANGLE X‐RAY SCATTERING STUDIES (WAXS)

    Energy Technology Data Exchange (ETDEWEB)

    Sangappa,, E-mail: sangappa@mangaloreuniversity.ac.in; Asha, S, E-mail: sangappa@mangaloreuniversity.ac.in [Department of Studies in Physics, Mangalore University, Mangalagangotri‐574 199 (India); Sanjeev, Ganesh, E-mail: sangappa@mangaloreuniversity.ac.in [Microtron Center, Mangalore University, Mangalagangotri‐574 199 (India); Subramanya, G, E-mail: sangappa@mangaloreuniversity.ac.in [Department of Studies in Sericulture, University of Mysore, Manasagangotri, Mysore‐570 006 (India); Parameswara, P, E-mail: sangappa@mangaloreuniversity.ac.in; Somashekar, R, E-mail: sangappa@mangaloreuniversity.ac.in [Department of Studies in Physics, University of Mysore, Manasagangotri, Mysore‐570 006 (India)

    2010-01-05

    The present work looks into the microstructural modification in electron irradiated Bombyx mori P31 silk fibers. The irradiation process was performed in air at room temperature using 8 MeV electron accelerator at different doses: 0, 25, 50 and 100 kGy. Irradiation of polymer is used to cross‐link or degrade the desired component or to fix the polymer morphology. The changes in microstructural parameters in these natural polymer fibers have been computed using wide angle X‐ray scattering (WAXS) data and employing line profile analysis (LPA) using Fourier transform technique of Warren. Exponential, Lognormal and Reinhold functions for the column length distributions have been used for the determination of crystal size, lattice strain and enthalpy parameters.

  12. Influência de genótipos de amoreira (Morus sp. e substratos no peso e características de casulos do bicho-da-seda (Bombyx mori L. Influence of mulberry (Morus sp. genotypes and substrates in weight and characteristics of silkworm cocoons (Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    Odinete Murari

    2001-05-01

    Full Text Available Avaliou-se o efeito de três genótipos de amoreira, Morus sp. (Moraceae: Miura, FM Shima Miura e IZ 56/4, três tipos de esteiras de criação: terra compactada, concreto e tela plástica sobre o peso e algumas características industriais de casulos produzidos pelo bicho-da-seda, Bombyx mori (Lepidoptera. Houve influência das interações de genótipos com esteiras de criação sobre o peso dos casulos produzidos. Com relação às características industriais, os tratamentos que mais se sobressaíram foram: Miura / terra compactada, FM-SM / tela plástica, IZ 56/4 / terra compactada e IZ 56/4 / tela plástica.The effect of three genotypes of mulberry, Morus sp. (Moraceae, namely, Miura, FM Shima Miura and IZ 56/4, and three types of rearing substrate comprising compact soil, concrete and plastic screen were estimated on weight and on certain industrial characteristics of Bombyx mori (Lepidoptera cocoons. Genotype interactions with rearing substrates affected weight of cocoons produced. Analysis showed the best treatments for manufacturer parameters were Miura / compact soil, FM-Shima Miura / plastic screen, IZ 56/4 / compact soil and IZ 56/4 / plastic screen.

  13. Insulin and 20-hydroxyecdysone action in Bombyx mori: Glycogen content and expression pattern of insulin and ecdysone receptors in fat body.

    Science.gov (United States)

    Keshan, Bela; Thounaojam, Bembem; Kh, Sanathoibi D

    2017-01-15

    Insulin and ecdysone signaling play a critical role on the growth and development of insects including Bombyx mori. Our previous study showed that Bombyx larvae reached critical weight for metamorphosis between day 3.5 and 4 of the fifth larval instar. The present study showed that the effect of insulin on the accumulation of glycogen in fat body of Bombyx larvae depends on the critical growth period. When larvae are in active growth period (before reaching critical weight), insulin caused increased accumulation of glycogen, while its treatment in larvae at terminal growth period (after critical period) resulted in an increased mobilization of glycogen. During terminal growth period, insulin and 20-hydroxyecdysone (20E) showed an antagonistic effect on the accumulation of fat body glycogen in fed, food deprived and decapitated larvae as well as in isolated abdomens. Insulin treatment decreased the glycogen content, whereas, 20E increased it. Food deprivation and decapitation caused an increase in the transcript levels of insulin receptor (InR) and this increase in InR expression might be attributed to a decrease in synthesis/secretion of insulin-like peptides, as insulin treatment in these larvae showed a down-regulation in InR expression. However, insulin showed an up-regulation in InR in isolated abdomens and it suggests that in food deprived and decapitated larvae, the exogenous insulin may interact with some head and/or thoracic factors in modulating the expression of InR. Moreover, in fed larvae, insulin-mediated increase in InR expression indicates that its regulation by insulin-like peptides also depends on the nutritional status of the larvae. The treatment of 20E in fed larvae showed an antagonistic effect on the transcript levels since a down-regulation in InR expression was observed. 20E treatment also led to a decreased expression of InR in food deprived and decapitated larvae as well as in isolated abdomens. Insulin and 20E also modulated the

  14. Microstructural parameters in 8 MeV Electron irradiated Bombyx mori silk fibers by wide-angle X-ray scattering studies (WAXS)

    International Nuclear Information System (INIS)

    Halabhavi, Sangappa

    2009-01-01

    The present work looks into the microstructural modification in Bombyx mori silk fibers, induced by electron irradiation. The irradiation process was performed in air at room temperature by use of 8 MeV electron accelerators at different doses: 0, 25, 50, 75 and 100 kGy respectively. Irradiation of polymer can be used to crosslink or degrade the desired component or to fixate the polymer morphology. The changes in microstructural parameters in these natural polymer fibers have been studied using wide angle X-ray scattering (WAXS) method. The crystal imperfection parameters such as crystallite size , lattice strain (g in %) and enthalpy (a * ) have been determined by line profile analysis (LPA) using Fourier method of Warren. Exponential, Lognormal and Reinhold functions for the column length distributions have been used for the determination of these parameters. The goodness of the fit and the consistency of these results suggest that the exponential distribution gives much better results, even though lognormal distribution has been widely used to estimate the similar stacking faults in metal oxide compounds. (author)

  15. A novel hAT element in Bombyx mori and Rhodnius prolixus: its relationship with miniature inverted repeat transposable elements (MITEs) and horizontal transfer.

    Science.gov (United States)

    Zhang, H-H; Shen, Y-H; Xu, H-E; Liang, H-Y; Han, M-J; Zhang, Z

    2013-10-01

    Comparative analysis of transposable elements (TEs) from different species can make it possible to reconstruct their history over evolutionary time. In this study, we identified a novel hAT element in Bombyx mori and Rhodnius prolixus with characteristic GGGCGGCA repeats in its subterminal region. Meanwhile, phylogenetic analysis demonstrated that the elements in these two species might represent a separate cluster of the hAT superfamily. Strikingly, a previously identified miniature inverted repeat transposable element (MITE) shared high identity with this autonomous element across the entire length, supporting the hypothesis that MITEs are derived from the internal deletion of DNA transposons. Interestingly, identity of the consensus sequences of this novel hAT element between B. mori and R. prolixus, which diverged about 370 million years ago, was as high as 96.5% over their full length (about 3.6 kb) at the nucleotide level. The patchy distribution amongst species, coupled with overall lack of intense purifying selection acting on this element, suggest that this novel hAT element might have experienced horizontal transfer between the ancestors of B. mori and R. prolixus. Our results highlight that this novel hAT element could be used as a potential tool for germline transformation of R. prolixus to control the transmission of Trypanosoma cruzi, which causes Chagas disease. © 2013 Royal Entomological Society.

  16. A cadherin-like protein functions as a receptor for Bacillus thuringiensis Cry1Aa and Cry1Ac toxins on midgut epithelial cells of Bombyx mori larvae.

    Science.gov (United States)

    Hara, Hirotaka; Atsumi, Shogo; Yaoi, Katsuro; Nakanishi, Kazuko; Higurashi, Satoshi; Miura, Nami; Tabunoki, Hiroko; Sato, Ryoichi

    2003-03-13

    Aminopeptidase N (APN) and cadherin-like protein (BtR175) from Bombyx mori larvae were examined for their roles in Cry1Aa- and Cry1Ac-induced lysis of B. mori midgut epithelial cells (MECs). APNs and BtR175 were present in all areas of the midgut, were particularly abundant in the posterior region, and were found only on columnar cell microvilli and not on the lateral membrane that makes cell-cell contacts. This distribution was in accordance with the distribution of Cry1A-susceptible MECs in the midgut. The lytic activity of Cry1Aa and Cry1Ac on collagenase-dissociated MECs was linearly dependent on toxin concentration. Although pre-treatment of MECs with anti-BtR175 antibody was observed to partially inhibit the lytic activity exerted by 0.1-1 nM Cry1Aa toxin or 5 nM Cry1Ac toxin, no significant inhibition was observed when MECs were pre-treated with anti-APN antibody. These results suggest that BtR175 functions as a major receptor for Cry1A toxins in the midgut of B. mori larvae.

  17. Expression of a sugar clade gustatory receptor, BmGr6, in the oral sensory organs, midgut, and central nervous system of larvae of the silkworm Bombyx mori.

    Science.gov (United States)

    Mang, Dingze; Shu, Min; Endo, Haruka; Yoshizawa, Yasutaka; Nagata, Shinji; Kikuta, Shingo; Sato, Ryoichi

    2016-03-01

    Insects taste nonvolatile chemicals through gustatory receptors (Grs) and make choices for feeding, mating, and oviposition. To date, genome projects have identified 69 Gr genes in the silkworm, Bombyx mori; however, the expression sites of these Grs remain to be explored. In this study, we used reverse transcription (RT)-PCR to investigate expression of the B. mori Gr-6 (BmGr6) gene, a member of the putative sugar clade gene family in various tissues. BmGr6 is expressed in the midgut, central nervous system (CNS), and oral sensory organs. Moreover, immunohistochemistry using an anti-BmGr6 antiserum demonstrated that BmGr6 is expressed in cells by oral sensory organs, midgut and nervous system. Furthermore, double-immunohistochemistry indicated that BmGr6 is expressed in midgut enteroendocrine cells, also in CNS neurosecretory cells. In particular, a portion of BmGr6-expressing cells, in both midgut and CNS, secretes FMRFamide-related peptides (FaRPs). These results suggest that BmGr6 functions not only as a taste receptor, but also as a chemical sensor such as for the regulation of gut movement, physiological conditions, and feeding behavior of larvae. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Immobilization of bioactive fibroblast growth factor-2 into cubic proteinous microcrystals (Bombyx mori cypovirus polyhedra) that are insoluble in a physiological cellular environment.

    Science.gov (United States)

    Mori, Hajime; Shukunami, Chisa; Furuyama, Akiko; Notsu, Hiroyuki; Nishizaki, Yuriko; Hiraki, Yuji

    2007-06-08

    The supramolecular architecture of the extracellular matrix and the disposition of its specific accessory molecules give rise to variable heterotopic signaling cues for single cells. Here we have described the successful occlusion of human fibroblast growth factor-2 (FGF-2) into the cubic inclusion bodies (FGF-2 polyhedra) of the Bombyx mori cytoplasmic polyhedrosis virus (BmCPV). The polyhedra are proteinous cubic crystals of several microns in size that are insoluble in the extracellular milieu. Purified FGF-2 polyhedra were found to stimulate proliferation and phosphorylation of p44/p42 mitogen-activated protein kinase in cultured fibroblasts. Moreover, cellular responses were blocked by a synthetic inhibitor of the FGF signaling pathway, SU5402, suggesting that FGF-2 polyhedra indeed act through FGF receptors. Furthermore, FGF-2 polyhedra retained potent growth stimulatory properties even after desiccation. We have demonstrated that BmCPV polyhedra microcrystals that occlude extracellular signaling proteins are a novel and versatile tool that can be employed to analyze cellular behavior at the single cell level.

  19. New insights into the genomic organization and splicing of the doublesex gene, a terminal regulator of sexual differentiation in the silkworm Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Jianping Duan

    Full Text Available Sex-determination mechanisms differ among organisms. The primary mechanism is diverse, whereas the terminal regulator is relatively-conserved. We analyzed the transcripts of the Bombyx mori doublesex gene (Bmdsx, and reported novel results concerning the genomic organization and expression of Bmdsx. Bmdsx consists of nine exons and eight introns, of which two exons are novel and have not been reported previously. Bmdsx transcripts are spliced to generate seventeen alternatively-spliced forms and eleven putative trans-spliced variants. Thirteen of the alternatively-spliced forms and five of the putative trans-spliced forms are reported here for the first time. Sequence analysis predicts that ten female-specific, six male-specific splice forms and one splice form found in males and females will result in four female-specific, two male-specific Dsx proteins and one Dsx protein common to males and females. The Dsx proteins are expected to be functional and regulate downstream target genes. Some of the predicted Dsx proteins are described here for the first time. Therefore the expression of the dsx gene in B. mori results in a variety of cis- and trans-spliced transcripts and multiple Dsx proteins. These findings show that in B. mori there is a complicated pattern of dsx splicing, and that the regulation of splicing and sex-specific functions of lepidopteran dsx have evolved complexity.

  20. Heterologous expression, purification and characterization of human β-1,2-N-acetylglucosaminyltransferase II using a silkworm-based Bombyx mori nucleopolyhedrovirus bacmid expression system.

    Science.gov (United States)

    Miyazaki, Takatsugu; Kato, Tatsuya; Park, Enoch Y

    2018-02-03

    β-1,2-N-Acetylglucosaminyltransferase II (GnTII, EC 2.4.1.143) is a Golgi-localized type II transmembrane enzyme that catalyzes the transfer of N-acetylglucosamine to the 6-arm of the trimanosyl core of N-glycans, an essential step in the conversion of oligomannose-type to complex-type N-glycans. Despite its physiological importance, there have been only a few reports on the heterologous expression and structure-function relationship of this enzyme. Here, we constructed a silkworm-based Bombyx mori nucleopolyhedrovirus bacmid expression system and expressed human GnTII (hGnTII) lacking the N-terminal cytosolic tail and transmembrane region. The recombinant hGnTII was purified from silkworm larval hemolymph in two steps by using tandem affinity purification tags, with a yield of approximately 120 μg from 10 mL hemolymph, and exhibited glycosyltransferase activity and strict substrate specificity. The enzyme was found to be N-glycosylated by the enzymatic cleavage of glycans, while hGnTII expressed in insect cells had not been reported to be glycosylated. Although insects typically produce pauci-mannosidic-type glycans, the structure of N-glycans in the recombinant hGnTII was suggested to be of the complex type, and the removal of the glycans did not affect the enzymatic activity. Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  1. Evidence for growth of strains of the plant epiphytic bacterium Erwinia herbicola and transconjugation among the bacterial strains in guts of the silkworm Bombyx mori.

    Science.gov (United States)

    Watanabe, K; Hara, W; Sato, M

    1998-09-01

    Growth of plant epiphytic bacteria Erwinia herbicola and Pseudomonas syringae in guts of the silkworm, Bombyx mori, was studied. Fifth instar silkworm larvae were fed artificial diets supplemented with these bacteria for 6 to 12 h followed by uncontaminated diets. At 1, 3, and 6 days after feeding, bacteria were isolated from insect guts and feces. A much larger population of E. herbicola was detected in the samples collected 3 and 6 days after the inoculation than in samples collected after 1 day, indicating that these bacteria grew in the insect gut, while P. syringae was unable to survive. Transconjugation between E. herbicola strains in the insect gut was also examined. First, either a donor or a recipient strain was fed to the insects in artificial diets containing the bacteria during 12 h, and then pairing strains were fed during 12 h after starvation for 12 h. The conjugative plasmid pBPW1::Tn7 was transferred into recipient cells at very high frequencies (10(-1)/recipient after 3 days and 10(-3) after 6 days) in insect guts. Indigenous plasmids of E. herbicola mobilized RSF1010 plasmid into recipient cells at frequencies of 10(-4) in insect guts. These transconjugants were detected in the feces of the insects. Thus, plasmid-mediated gene transfer among the epiphytic bacteria in insect guts was demonstrated. The results obtained suggest that in insecta gene transfer may play an important role in the evolution of plant epiphytic bacteria. Copyright 1998 Academic Press.

  2. Effects of Starvation and Thermal Stress on the Thermal Tolerance of Silkworm, Bombyx mori: Existence of Trade-offs and Cross-Tolerances.

    Science.gov (United States)

    Mir, A H; Qamar, A

    2017-09-27

    Organisms, in nature, are often subjected to multiple stressors, both biotic and abiotic. Temperature and starvation are among the main stressors experienced by organisms in their developmental cycle and the responses to these stressors may share signaling pathways, which affects the way these responses are manifested. Temperature is a major factor governing the performance of ectothermic organisms in ecosystems worldwide and, therefore, the thermal tolerance is a central issue in the thermobiology of these organisms. Here, we investigated the effects of starvation as well as mild heat and cold shocks on the thermal tolerance of the larvae of silkworm, Bombyx mori (Linnaeus). Starvation acted as a meaningful or positive stressor as it improved cold tolerance, measured as chill coma recovery time (CCRT), but, at the same time, it acted as a negative stressor and impaired the heat tolerance, measured as heat knockdown time (HKT). In the case of heat tolerance, starvation negated the positive effects of both mild cold as well as mild heat shocks and thus indicated the existence of trade-off between these stressors. Both mild heat and cold shocks improved the thermal tolerance, but the effects were more prominent when the indices were measured in response to a stressor of same type, i.e., a mild cold shock improved the cold tolerance more than the heat tolerance and vice versa. This improvement in thermal tolerance by both mild heat as well as cold shocks indicated the possibility of cross-tolerance between these stressors.

  3. Rapid detection of Bombyx mori nucleopolyhedrovirus (BmNPV) by loop-mediated isothermal amplification assay combined with a lateral flow dipstick method.

    Science.gov (United States)

    Zhou, Yang; Wu, Jiege; Lin, Feng; Chen, Naifu; Yuan, Shaofei; Ding, Lina; Gao, Li; Hang, Bangxing

    2015-12-01

    The Bombyx mori nucleopolyhedrovirus (BmNPV) is a principal pathogen of the domestic silkworm. The disease often breaks out in sericultural countries and due to its high infectivity; it is difficult to control, resulting in heavy economic loss. In order to develop a rapid, sensitive visual detection and simple-to-use novel technology for detection of BmNPV, a loop-mediated isothermal amplification (LAMP) assay combined with a lateral flow dipstick (LFD) method was described. In this study, a set of four primers and a labeled probe were designed specifically to recognize six distinct regions of the BmNPV gp41 gene, and the LAMP for the detection of BmNPV was developed by isothermal amplification at 61 °C for 45 min, followed by hybridization with an FITC-labeled DNA probe for 5 min and detected by LFD within 5 min. The detection limit of LAMP-LFD was 0.2 pg DNA extracted from silkworm infected with BmNPV and was 100 times more sensitive than conventional PCR. No product was generated from silkworm infected with other viruses. Furthermore, we applied the technique to detect BmNPV in the hemolymph and feces at different intervals post infection (pi). In conclusion, the novel LAMP-LFD setup presented here is simple, rapid, reliable, and has the potential for future use in the detection of BmNPV. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Characterization of the Gene BmEm4, a Homologue of Drosophila E(splm4, from the Silkworm, Bombyx mori

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    Fenghui Zeng

    2009-01-01

    Full Text Available The Drosophila E(splm4 gene contains some highly conserved motifs (such as the Brd box, GY box, K box, and CAAC motif in its 3′ untranslated region (3′UTR. It was shown to be a microRNA target gene in Drosophila and to play an important role in the regulation of neurogenesis. We identified a homologue of the E(splm4 gene from Bombyx mori called BmEm4 and examined the expression patterns of BmEm4 mRNA and protein. There was a lack of correlation in the expression of the mRNA and protein between the different developmental stages, which raises the possibility of posttranscriptional regulation of the BmEm4 mRNA. Consistent with this idea is the finding that the 3′ UTR contains two putative binding sites for microRNAs. Moreover, given that the expression is the highest in the larval head, as confirmed by immunohistochemistry, we propose that BmEm4 may also be involved in the regulation of neurogenesis. Immunostaining indicated that BmEm4 is located primarily in the cytoplasm.

  5. Characterization of the Gene BmEm4, a Homologue of Drosophila E(spl)m4, from the Silkworm, Bombyx mori

    Science.gov (United States)

    Zeng, Fenghui; Xie, Hongxia; Nie, Zuoming; Chen, Jian; Lv, Zhengbing; Chen, Jianqing; Wang, Dan; Liu, Lili; Yu, Wei; Sheng, Qing; Wu, Xiangfu; Zhang, Yaozhou

    2009-01-01

    The Drosophila E(spl)m4 gene contains some highly conserved motifs (such as the Brd box, GY box, K box, and CAAC motif) in its 3′ untranslated region (3′ UTR). It was shown to be a microRNA target gene in Drosophila and to play an important role in the regulation of neurogenesis. We identified a homologue of the E(spl)m4 gene from Bombyx mori called BmEm4 and examined the expression patterns of BmEm4 mRNA and protein. There was a lack of correlation in the expression of the mRNA and protein between the different developmental stages, which raises the possibility of posttranscriptional regulation of the BmEm4 mRNA. Consistent with this idea is the finding that the 3′ UTR contains two putative binding sites for microRNAs. Moreover, given that the expression is the highest in the larval head, as confirmed by immunohistochemistry, we propose that BmEm4 may also be involved in the regulation of neurogenesis. Immunostaining indicated that BmEm4 is located primarily in the cytoplasm. PMID:19830255

  6. Comparative Transcriptome Analysis of Bombyx mori (Lepidoptera Larval Midgut Response to BmNPV in Susceptible and Near-Isogenic Resistant Strains.

    Directory of Open Access Journals (Sweden)

    Xue-Yang Wang

    Full Text Available Bombyx mori nucleopolyhedrovirus (BmNPV is one of the primary pathogens causing severe economic losses in sericulture. However, the molecular mechanism of silkworm resistance to BmNPV remains largely unknown. Here, the recurrent parent P50 (susceptible strain and the near-isogenic line BC9 (resistance strain were used in a comparative transcriptome study examining the response to infection with BmNPV. A total of 14,300 unigenes were obtained from two different resistant strains; of these, 869 differentially expressed genes (DEGs were identified after comparing the four transcriptomes. Many DEGs associated with protein metabolism, cytoskeleton, and apoptosis may be involved in the host response to BmNPV infection. Moreover, some immunity related genes were also altered following BmNPV infection. Specifically, after removing genetic background and individual immune stress response genes, 22 genes were found to be potentially involved in repressing BmNPV infection. These genes were related to transport, virus replication, intracellular innate immune, and apoptosis. Our study provided an overview of the molecular mechanism of silkworm resistance to BmNPV infection and laid a foundation for controlling BmNPV in the future.

  7. Transgenic Clustered Regularly Interspaced Short Palindromic Repeat/Cas9-Mediated Viral Gene Targeting for Antiviral Therapy of Bombyx mori Nucleopolyhedrovirus.

    Science.gov (United States)

    Chen, Shuqing; Hou, Chengxiang; Bi, Honglun; Wang, Yueqiang; Xu, Jun; Li, Muwang; James, Anthony A; Huang, Yongping; Tan, Anjiang

    2017-04-15

    We developed a novel antiviral strategy by combining transposon-based transgenesis and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system for the direct cleavage of Bombyx mori nucleopolyhedrovirus (BmNPV) genome DNA to promote virus clearance in silkworms. We demonstrate that transgenic silkworms constitutively expressing Cas9 and guide RNAs targeting the BmNPV immediate early-1 ( ie-1 ) and me53 genes effectively induce target-specific cleavage and subsequent mutagenesis, especially large (∼7-kbp) segment deletions in BmNPV genomes, and thus exhibit robust suppression of BmNPV proliferation. Transgenic animals exhibited higher and inheritable resistance to BmNPV infection than wild-type animals. Our approach will not only contribute to modern sericulture but also shed light on future antiviral therapy. IMPORTANCE Pathogen genome targeting has shown its potential in antiviral research. However, transgenic CRISPR/Cas9 system-mediated viral genome targeting has not been reported as an antiviral strategy in a natural animal host of a virus. Our data provide an effective approach against BmNPV infection in a real-world biological system and demonstrate the potential of transgenic CRISPR/Cas9 systems in antiviral research in other species. Copyright © 2017 Chen et al.

  8. SWATH-based quantitative proteomics reveals the mechanism of enhanced Bombyx mori nucleopolyhedrovirus-resistance in silkworm reared on UV-B treated mulberry leaves.

    Science.gov (United States)

    Hu, Jin; Zhu, Wei; Li, Yaohan; Guan, Qijie; Yan, Haijian; Yu, Jiaojiao; Fu, Zhirong; Lu, Xingmeng; Tian, Jingkui

    2017-07-01

    Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the most acute infectious diseases in silkworm, which has led to great economic loss in sericulture. Previous study showed that the content of secondary metabolites in mulberry leaves, particularly for moracin N, was increased after UV-B irradiation. In this study, the BmNPV resistance of silkworms reared on UV-B treated and moracin N spread mulberry leaves was improved. To uncover the mechanism of enhanced BmNPV resistance, silkworm midguts from UV-B treated mulberry leaves (BUM) and moracin N (BNM) groups were analyzed by SWATH-based proteomic technique. Of note, the abundance of ribosomal proteins in BUM and BNM groups was significantly changed to maintain the synthesis of total protein levels and cell survival. While, cytochrome c oxidase subunit II, calcium ATPase and programmed cell death 4 involved in apoptotic process were up-regulated in BNM group. Expressions of lipase-1, serine protease precursor, Rab1 protein, and histone genes were increased significantly in BNM group. These results suggest that moracin N might be the main active component in UV-B treated mulberry leaves which could improve the BmNPV-resistance of silkworm through promoting apoptotic cell death, enhancing the organism immunity, and regulating the intercellular environment of cells in silkworm. It also presents an innovative process to reduce the mortality rate of silkworms infected with BmNPV. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Expression of Trichoderma viride endoglucanase III in the larvae of silkworm, Bombyx mori L. and characteristic analysis of the recombinant protein.

    Science.gov (United States)

    Li, Xing-hua; Zhang, Peng; Wang, Mei-xian; Zhou, Fang; Malik, Firdose Ahmad; Yang, Hua-jun; Bhaskar, Roy; Hu, Jia-biao; Sun, Chun-guang; Miao, Yun-gen

    2011-08-01

    Endoglucanase is a part of cellulase which hydrolyzes cellulose into glucose. In this study, we cloned endoglucanase III (EG III) gene from Trichoderma viride strain AS 3.3711 using a PCR-based exon splicing method, and expressed EG III recombinant protein in both silkworm BmN cell line and silkworm larvae with an improved Bac-to-Bac/BmNPV mutant baculovirus expression system, which lacks the chiA and v-cath genes of Bombyx mori nucleopolyhedrovirus (BmNPV). The result showed that around 45 kDa protein was visualized in BmN cells at 48 h after the second generation recombinant mBacmid/BmNPV/EG III baculovirus infection. The enzymes from recombinant baculoviruses infected silkworms exhibited significant maximum enzyme activity at the environmental condition of pH 8.0 and temperature 50°C, and increased 20.94 and 19.13% compared with that from blank mBacmid/BmNPV baculoviruses infected silkworms and normal silkworms, respectively. It was stable at pH range from 5.0 to 9.0 and at temperature range from 40 to 60°C. It provided a possibility to generate transgenic silkworms expressing bio-active cellulase, which can catabolize dietary fibers more efficiently, and it might be of great significance for sericulture industry.

  10. A single amino acid substitution modulates low-pH-triggered membrane fusion of GP64 protein in Autographa californica and Bombyx mori nucleopolyhedroviruses

    International Nuclear Information System (INIS)

    Katou, Yasuhiro; Yamada, Hayato; Ikeda, Motoko; Kobayashi, Michihiro

    2010-01-01

    We have previously shown that budded viruses of Bombyx mori nucleopolyhedrovirus (BmNPV) enter the cell cytoplasm but do not migrate into the nuclei of non-permissive Sf9 cells that support a high titer of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) multiplication. Here we show, using the syncytium formation assay, that low-pH-triggered membrane fusion of BmNPV GP64 protein (Bm-GP64) is significantly lower than that of AcMNPV GP64 protein (Ac-GP64). Mutational analyses of GP64 proteins revealed that a single amino acid substitution between Ac-GP64 H155 and Bm-GP64 Y153 can have significant positive or negative effects on membrane fusion activity. Studies using bacmid-based GP64 recombinant AcMNPV harboring point-mutated ac-gp64 and bm-gp64 genes showed that Ac-GP64 H155Y and Bm-GP64 Y153H substitutions decreased and increased, respectively, the multiplication and cell-to-cell spread of progeny viruses. These results indicate that Ac-GP64 H155 facilitates the low-pH-triggered membrane fusion reaction between virus envelopes and endosomal membranes.

  11. Comparative Transcriptome Analysis of Bombyx mori (Lepidoptera) Larval Midgut Response to BmNPV in Susceptible and Near-Isogenic Resistant Strains.

    Science.gov (United States)

    Wang, Xue-Yang; Yu, Hai-Zhong; Geng, Lei; Xu, Jia-Ping; Yu, Dong; Zhang, Shang-Zhi; Ma, Yan; Fei, Dong-Qiong

    2016-01-01

    Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the primary pathogens causing severe economic losses in sericulture. However, the molecular mechanism of silkworm resistance to BmNPV remains largely unknown. Here, the recurrent parent P50 (susceptible strain) and the near-isogenic line BC9 (resistance strain) were used in a comparative transcriptome study examining the response to infection with BmNPV. A total of 14,300 unigenes were obtained from two different resistant strains; of these, 869 differentially expressed genes (DEGs) were identified after comparing the four transcriptomes. Many DEGs associated with protein metabolism, cytoskeleton, and apoptosis may be involved in the host response to BmNPV infection. Moreover, some immunity related genes were also altered following BmNPV infection. Specifically, after removing genetic background and individual immune stress response genes, 22 genes were found to be potentially involved in repressing BmNPV infection. These genes were related to transport, virus replication, intracellular innate immune, and apoptosis. Our study provided an overview of the molecular mechanism of silkworm resistance to BmNPV infection and laid a foundation for controlling BmNPV in the future.

  12. Inhibition of miR-274-3p increases BmCPV replication by regulating the expression of BmCPV NS5 gene in Bombyx mori.

    Science.gov (United States)

    Wu, Ping; Jiang, Xiaoxu; Sang, Qi; Annan, Enoch; Cheng, Tao; Guo, Xijie

    2017-08-01

    Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) is one of the major pathogens that pose a big challenge to the sericulture industry. Growing evidences have shown that microRNAs play key roles in the regulations of host-pathogen interactions in insects. MicroRNAs have been found in silkworms, whether and how they affect the silkworm-BmCPV interactions are still unknown. Here we investigate the effect of miR-274-3p on the BmCPV replication in the BmCPV-infected silkworm larvae. In our study, BmCPV Nonstructural protein 5 (NS5) was identified to be the target of miR-274-3p based on bioinformatics analysis and luciferase reporter assay. The abundance of NS5 was significantly increased in the presence of miR-274-3p inhibitor based on the qRT-PCR and Western blotting results. Further, qRT-PCR results revealed that the expression of polyhedrin gene of BmCPV in the larvae after applying miR-274-3p inhibitor was significantly increased comparing with that of larvae with negative control. Our results suggest that inhibition of miR-274-3p could facilitate BmCPV replication by up-regulating BmCPV NS5 gene expression and are insightful for further exploring the interactions between silkworm and BmCPV.

  13. Construction of an in vivo system for functional analysis of the genes involved in sex pheromone production in the silkmoth, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Ken-Ichi eMoto

    2012-02-01

    Full Text Available Moths produce species-specific sex pheromones to attract conspecific mates. The biochemical processes that comprise sex pheromone biosynthesis are precisely regulated and a number of gene products are involved in this biosynthesis and regulation. In recent years, at least 300 EST clones have been isolated from Bombyx mori pheromone gland (PG specific cDNA libraries with some of those clones (i.e., B. mori PG-specific desaturase1 (Bmpgdesat1, PG-specific fatty-acyl reductase (pgFAR, PG-specific acyl-CoA-binding protein (pgACBP, B. mori fatty acid transport protein (BmFATP, B. mori lipid storage droplet protein-1 (BmLsd1 characterized and demonstrated to play a role in sex pheromone production. However, most of the EST clones have yet to be fully characterized and identified. To develop an efficient system for analyzing sex pheromone production-related genes, we investigated the feasibility of a novel gene analysis system using the upstream region of Bmpgdesat1 that should contain a PG-specific gene promoter in conjunction with piggyBac vector-mediated germ-line transformation. As a result, we have been able to obtain expression of our reporter gene (enhanced green fluorescent protein in the PG but not in other tissues of transgenic B. mori. Current results indicate that we have successfully constructed a novel in vivo gene analysis system for sex pheromone production in B. mori.

  14. Estudios electrofisiológicos de estructura-actividad de la célula antenal receptora del ácido benzoico de la hembra de Bombyx mori L

    OpenAIRE

    de Brito Sanchez, María Gabriela

    2000-01-01

    El objetivo de este trabajo fue caracterizar las respuestas electrofisiológicas de la célula receptora olfativa al ácido benzoico, presentes en las hembras de la mariposa del gusano de la seda Bombyx mori. Nuestros estudios permiten concluir que la célula receptora al ácido benzoico es un receptor especialista ya que responde máximamente a su compuesto llave y tiene un espectro idéntico para compuestos menos efectivos. Mediante estudios de estructura-actividad con substituyentes halogenados d...

  15. Characterization of a novel C-type lectin, Bombyx mori multibinding protein, from the B. mori hemolymph: mechanism of wide-range microorganism recognition and role in immunity.

    Science.gov (United States)

    Watanabe, Ayako; Miyazawa, Sousui; Kitami, Madoka; Tabunoki, Hiroko; Ueda, Kenjiro; Sato, Ryoichi

    2006-10-01

    To investigate the system used by insects to recognize invading microorganisms, we examined proteins from the larval hemolymph of Bombyx mori that bind to the cell surface of microorganisms. Two hemolymph proteins that bound to the cell surfaces of Micrococcus luteus and Saccharomyces cerevisiae were shown to be identical. This protein bound to all 11 microorganisms examined-5 Gram-negative bacteria, 3 Gram-positive bacteria, and 3 yeasts-and was consequently designated B. mori multibinding protein (BmMBP). The sequence of the cDNA encoding BmMBP revealed that it was a C-type lectin with two dissimilar carbohydrate-recognition domains (CRD1 and CRD2) distantly related to known insect C-type lectins. CRD1 and CRD2 were prepared as recombinant proteins and their binding properties were investigated using inhibition assays. Each domain had wide, dissimilar binding spectra to sugars. These properties enable BmMBP to bind to two sites on a microorganism, facilitating high-affinity binding to many types of microorganisms. The dissociation constants of BmMBP with M. luteus cells and S. cerevisiae were 1.23 x 10(-8) and 1.00 x 10(-11) M, respectively. rBmMBP triggered the aggregation of hemocytes from B. mori larvae in vitro and microorganisms recognized by BmMBP were surrounded by aggregated hemocytes in vivo, forming a nodule, which is the typical cellular reaction in insect immune responses. These observations suggest that BmMBP functions as a trigger for the nodule reaction and that the multirecognition characteristic of BmMBP plays an important role in the early stages of infection by a variety of microorganisms.

  16. D181A Site-Mutagenesis Enhances Both the Hydrolyzing and Transfructosylating Activities of BmSUC1, a Novel β-Fructofuranosidase in the Silkworm Bombyx mori

    Directory of Open Access Journals (Sweden)

    Quan Gan

    2018-02-01

    Full Text Available β-fructofuranosidase (β-FFase belongs to the glycosyl-hydrolase family 32 (GH32, which can catalyze both the release of β-fructose from β-d-fructofuranoside substrates to hydrolyze sucrose and the synthesis of short-chain fructooligosaccharide (FOS. BmSuc1 has been cloned and identified from the silkworm Bombyx mori as a first animal type of β-FFase encoding gene. It was hypothesized that BmSUC1 plays an important role in the silkworm-mulberry adaptation system. However, there is little information about the enzymatic core sites of BmSUC1. In this study, we mutated three amino acid residues (D63, D181, and E234 that represent important conserved motifs for β-FFase activity in GH32 to alanine respectively by using site-directed mutagenesis. Recombinant proteins of three mutants and wild type BmSUC1 were obtained by using a Bac-to-Bac/BmNPV expression system and BmN cells. Enzymatic activity, kinetic properties, and substrate specificity of the four proteins were analyzed. High Performance Liquid Chromatography (HPLC was used to compare the hydrolyzing and transfructosylating activities between D181A and wtBmSUC1. Our results revealed that the D63A and E234A mutations lost activity, suggesting that D63 and E234 are key amino acid residues for BmSUC1 to function as an enzyme. The D181A mutation significantly enhanced both hydrolyzing and transfructosylating activities of BmSUC1, indicating that D181 may not be directly involved in catalyzation. The results provide insight into the chemical catalyzation mechanism of BmSUC1 in B. mori. Up-regulated transfructosylating activity of BmSUC1 could provide new ideas for using B. mori β-FFase to produce functional FOS.

  17. Analysis of expression and chitin-binding activity of the wing disc cuticle protein BmWCP4 in the silkworm, Bombyx mori.

    Science.gov (United States)

    Deng, Hui-Min; Li, Yong; Zhang, Jia-Ling; Liu, Lin; Feng, Qi-Li

    2016-12-01

    The insect exoskeleton is mainly composed of chitin filaments linked by cuticle proteins. When insects molt, the cuticle of the exoskeleton is renewed by degrading the old chitin and cuticle proteins and synthesizing new ones. In this study, chitin-binding activity of the wing disc cuticle protein BmWCP4 in Bombyx mori was studied. Sequence analysis showed that the protein had a conservative hydrophilic "R&R" chitin-binding domain (CBD). Western blotting showed that BmWCP4 was predominately expressed in the wing disc-containing epidermis during the late wandering and early pupal stages. The immunohistochemistry result showed that the BmWCP4 was mainly present in the wing disc tissues containing wing bud and trachea blast during day 2 of wandering stage. Recombinant full-length BmWCP4 protein, "R&R" CBD peptide (CBD), non-CBD peptide (BmWCP4-CBD - ), four single site-directed mutated peptides (M 1 , M 2 , M 3 and M 4 ) and four-sites-mutated peptide (M F ) were generated and purified, respectively, for in vitro chitin-binding assay. The results indicated that both the full-length protein and the "R&R" CBD peptide could bind with chitin, whereas the BmWCP4-CBD - could not bind with chitin. The single residue mutants M 1 , M 2 , M 3 and M 4 reduced but did not completely abolish the chitin-binding activity, while four-sites-mutated protein M F completely lost the chitin-binding activity. These data indicate that BmWCP4 protein plays a critical role by binding to the chitin filaments in the wing during larva-to-pupa transformation. The conserved aromatic amino acids are critical in the interaction between chitin and the cuticle protein. © 2015 Institute of Zoology, Chinese Academy of Sciences.

  18. Involvement of phosphorylation of adenosine 5'-monophosphate-activated protein kinase in PTTH-stimulated ecdysteroidogenesis in prothoracic glands of the silkworm, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Shi-Hong Gu

    Full Text Available In this study, we investigated inhibition of the phosphorylation of adenosine 5'-monophosphate-activated protein kinase (AMPK by prothoracicotropic hormone (PTTH in prothoracic glands of the silkworm, Bombyx mori. We found that treatment with PTTH in vitro inhibited AMPK phosphorylation in time- and dose-dependent manners, as seen on Western blots of glandular lysates probed with antibody directed against AMPKα phosphorylated at Thr172. Moreover, in vitro inhibition of AMPK phosphorylation by PTTH was also verified by in vivo experiments: injection of PTTH into day 7 last instar larvae greatly inhibited glandular AMPK phosphorylation. PTTH-inhibited AMPK phosphorylation appeared to be partially reversed by treatment with LY294002, indicating involvement of phosphatidylinositol 3-kinase (PI3K signaling. A chemical activator of AMPK (5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside, AICAR increased both basal and PTTH-inhibited AMPK phosphorylation. Treatment with AICAR also inhibited PTTH-stimulated ecdysteroidogenesis of prothoracic glands. The mechanism underlying inhibition of PTTH-stimulated ecdysteroidogenesis by AICAR was further investigated by determining the phosphorylation of eIF4E-binding protein (4E-BP and p70 ribosomal protein S6 kinase (S6K, two known downstream signaling targets of the target of rapamycin complex 1 (TORC1. Upon treatment with AICAR, decreases in PTTH-stimulated phosphorylation of 4E-BP and S6K were detected. In addition, treatment with AICAR did not affect PTTH-stimulated extracellular signal-regulated kinase (ERK phosphorylation, indicating that AMPK phosphorylation is not upstream signaling for ERK phosphorylation. Examination of gene expression levels of AMPKα, β, and γ by quantitative real-time PCR (qRT-PCR showed that PTTH did not affect AMPK transcription. From these results, it is assumed that inhibition of AMPK phosphorylation, which lies upstream of PTTH-stimulated TOR signaling, may play a role in

  19. The arginine residue within the C-terminal active core of Bombyx mori pheromone biosynthesis-activating neuropeptide (PBAN is essential for receptor binding and activation

    Directory of Open Access Journals (Sweden)

    Takeshi eKawai

    2012-03-01

    Full Text Available In most lepidopteran insects, the biosynthesis of sex pheromones is regulated by pheromone biosynthesis activating neuropeptide (PBAN. Bombyx mori PBAN (BomPBAN consists of 33 amino acid residues and contains a C-terminus FSPRLamide motif as the active core. Among neuropeptides containing the FXPRLamide motif, the arginine (Arg, R residue two positions from the C-terminus is highly conserved across several neuropeptides, which can be designated as RXamide peptides. The purpose of this study was to reveal the role of the Arg residue in the BomPBAN active core. We synthesized a ten-residue peptide corresponding to the C-terminal part of BomPBAN with a series of point mutants at the 2nd position (ie, Arg from the C-terminus, termed the C2 position, and measured their efficacy in stimulating Ca2+ influx in insect cells concomitantly expressing a fluorescent PBAN receptor chimera (PBANR-EGFP and loaded with the fluorescent Ca2+ indicator, Fura Red-AM. PBAN analogs with the C2 position replaced with alanine (Ala, A, aspartic acid (Asp, D, serine (Ser, S or L-2-aminooctanoic acid (Aoc decreased PBAN-like activity. RC2A (SKTRYFSPALamide and RC2D (SKTRYFSPDLamide had the lowest activity and could not inhibit the activity of PBAN C10 (SKTRYFSPRLamide. We also prepared Rhodamine Red-labeled PBAN analogs of the mutants and examined their ability to bind PBANR. In contrast to 100 nM Rhodamine Red-PBAN C10, none of the mutants at the same concentration exhibited PBANR binding. Taken together, our results demonstrate that the C2 Arg residue in BomPBAN is essential for PBANR binding and activation.

  20. Functional characterization of Bombyx mori nucleopolyhedrovirus late gene transcription and genome replication factors in the non-permissive insect cell line SF-21

    International Nuclear Information System (INIS)

    Berretta, Marcelo F.; Deshpande, Mandar; Crouch, Erin A.; Passarelli, A. Lorena

    2006-01-01

    We compared the abilities of late gene transcription and DNA replication machineries of the baculoviruses Autographa californica nucleopolyhedrovirus (AcMNPV) and Bombyx mori NPV (BmNPV) in SF-21 cells, an insect-derived cell line permissive for AcMNPV infection. It has been well established that 19 AcMNPV late expression factors (lefs) stimulate substantial levels of late gene promoter activity in SF-21 cells. Thus, we constructed a set of clones containing the BmNPV homologs of the AcMNPV lefs under control of the constitutive Drosophila heat shock 70 protein promoter and tested their ability to activate an AcMNPV late promoter-reporter gene cassette in SF-21 cells. We tested the potential of individual or predicted functional groups of BmNPV lefs to successfully replace the corresponding AcMNPV gene(s) in transient late gene expression assays. We found that most, but not all, BmNPV lefs were able to either fully or partially substitute for the corresponding AcMNPV homolog in the context of the remaining AcMNPV lefs with the exception of BmNPV p143, ie-2, and p35. BmNPV p143 was unable to support late gene expression or be imported into the nucleus of cells in the presence of the AcMNPV or the BmNPV LEF-3, a P143 nuclear shuttling factor. Our results suggest that host-specific factors may affect the function of homologous proteins

  1. Bombyx mori DNA/RNA non-specific nuclease: expression of isoforms in insect culture cells, subcellular localization and functional assays.

    Science.gov (United States)

    Liu, Jisheng; Swevers, Luc; Iatrou, Kostas; Huvenne, Hanneke; Smagghe, Guy

    2012-08-01

    A DNA/RNA non-specific alkaline nuclease (BmdsRNase) was isolated from the digestive juice of Bombyx mori. While originally reported to be produced by the midgut only, in this project it was found that the mRNA of this enzyme was also expressed in the epidermis, fat body, gut, thoracic muscles, Malpighian tubules, brain, and silk glands of 5th instar larvae, indicating additional functions to its reported role in nucleic acid digestion in the midgut. In order to study the functional properties of BmdsRNase, three pEA-BmdsRNase expression constructs were generated, characterized by presence or absence of a signal peptide and a propeptide, and used for expression in lepidopteran Hi5 tissue culture cells. Western blot indicated that these different forms of BmdsRNase protein were not secreted into the growth medium, while they were detected in the pellets and supernatants of Hi5 cell extracts. Nucleic acids cleavage experiments indicated that full-length BmdsRNase could digest dsRNA and that the processed form (absence of signal peptide and propeptide) of BmdsRNase could degrade both DNA and dsRNA in Hi5 cell culture. Using a reporter assay targeted by transfected homologous dsRNA, it was shown that the digestive property of the processed form could interfere with the RNAi response. Immunostaining of processed BmdsRNase protein showed asymmetric localization in the cellular cytoplasm and co-localization with Flag-tagged Dicer-2 was also observed. In conclusion, our in vitro studies indicated that intracellular protein isoforms of BmdsRNase can be functional and involved in the regulation of nucleic acid metabolism in the cytoplasm. In particular, because of its propensity to degrade dsRNA, the enzyme might be involved in the innate immune response against invading nucleic acids such as RNA viruses. Copyright © 2012 Elsevier Ltd. All rights reserved.

  2. The Structural Variation Is Associated with the Embryonic Lethality of a Novel Red Egg Mutant Fuyin-lre of Silkworm, Bombyx mori.

    Science.gov (United States)

    Chen, Anli; Liao, Pengfei; Li, Qiongyan; Zhao, Qiaoling; Yang, Weike; Zhu, Shuifen; Wu, Fang; He, Rongfan; Dong, Zhanpeng; Huang, Ping

    2015-01-01

    Bombyx mori presents several types of egg color mutations, all of which have been extensively discussed in sericulture. While the red egg mutation has been previously observed, lethal red-egg mutants have not been reported. In the present work, the red egg mutant Fuyin-lre (Fuyin-lethal red egg) was discovered from the Fuyin germplasm resource of B. mori. This mutant features red-colored eggs and embryonic lethality. Genetic analysis showed that Fuyin-lre follows recessive inheritance, with the red egg gene re governing the egg color, and the embryonic lethality of Fuyin-lre may be caused by mutations of other genes closely linked to re. Digital gene expression (DGE) was employed to compare the transcription profiles of Fuyin and Fuyin-lre eggs after 24 and 48 h of incubation. A total of 48 differentially expressed genes followed the same expression patterns in both groups at both time points (FDR < 0.01 and log 2 Ratio ≥ 1). Further analyses indicated that 8 out of the 48 genes (including re) were closely linked to re. These 8 genes were highly expressed in wild-type Fuyin and the red egg mutant re but showed nearly absent expression in Fuyin-lre. Sequencing of the re gene confirmed that the re gene itself does not induce embryonic lethality, and structure analysis showed that the structural variation of the region where the 8 genes were located may be associated with the embryonic lethality of Fuyin-lre. The present work provides a good foundation for future studies on the mechanism of embryonic lethality and embryonic development in Fuyin-lre.

  3. Molecular mechanisms of silk gland damage caused by phoxim exposure and protection of phoxim-induced damage by cerium chloride in Bombyx mori.

    Science.gov (United States)

    Li, Bing; Sun, Qingqing; Yu, Xiaohong; Xie, Yi; Hong, Jie; Zhao, Xiaoyang; Sang, Xuezi; Shen, Weide; Hong, Fashui

    2015-09-01

    It is known that exposure to organophosphorus pesticides (OP) including phoxim can produce oxidative stress, neurotoxicity, and greatly attenuate cocooning rate in the silkworm, Bombyx mori. Cerium treatment has been demonstrated to relieve phoxim-induced toxicity in B. mori; however, very little is known about the molecular mechanisms of silk gland injury due to OP exposure and protection of gland damage due to cerium pretreatment. The aim of this study was to evaluate silk gland damage and its molecular mechanisms in phoxim-induced silkworm toxicity and the protective mechanisms of cerium following exposure to phoxim. The results showed that phoxim exposure resulted in severe gland damage, reductions in protein synthesis and the cocooning rate of silkworms. Cerium (Ce) attenuated gland damage caused by phoxim, promoted protein synthesis, increased the antioxidant capacity of the gland and increased the cocooning rate of B. mori. Furthermore, digital gene expression data suggested that phoxim exposure led to significant up-regulation of 714 genes and down-regulation of 120 genes. Of these genes, 122 were related to protein metabolism, specifically, the down-regulated Ser2, Ser3, Fib-L, P25, and CYP450. Ce pretreatment resulted in up-regulation of 162 genes, and down-regulation of 141 genes, importantly, Ser2, Ser3, Fib-L, P25, and CYP333B8 were up-regulated. Treatment with CeCl3 + phoxim resulted in higher levels of Fib-L, P25, Ser2, Ser3, CAT, TPx, and CYP333B8 expression in the silk gland of silkworms. These findings indicated that Ce increased cocooning rate via the promotion of silk protein synthesis-related gene expression in the gland under phoxim-induced toxicity. These findings may expand the application of rare earths in sericulture. © 2014 Wiley Periodicals, Inc.

  4. Differentially expressed genes in the head of the 2nd instar pre-molting larvae of the nm2 mutant of the silkworm, Bombyx mori

    Science.gov (United States)

    Wu, Fan; Xu, Pingzhen; Shen, Xingjia

    2017-01-01

    Molting is an important physiological process in the larval stage of Bombyx mori and is controlled by various hormones and peptides. The silkworm mutant that exhibits the phenotype of non-molting in the 2nd instar (nm2) is incapable of molting in the 2nd instar and dies after seven or more days. The ecdysone titer in the nm2 mutant is lower than that in the wildtype, and the mutant can be rescued by feeding with 20E and cholesterol. The results of positional cloning indicated that structural alteration of BmCPG10 is responsible for the phenotype of the nm2 mutant. To explore the possible relationship between BmCPG10 and the ecdysone titer as well as the genes affected by BmCPG10, digital gene expression (DGE) profile analysis was conducted in the nm2 mutant, with the wildtype strain C603 serving as the control. The results revealed 1727 differentially expressed genes, among which 651 genes were upregulated and 1076 were downregulated in nm2. BLASTGO analysis showed that these differentially expressed genes were involved in various biological processes, cellular components and molecular functions. KEGG analysis indicated an enrichment of these differentially expressed genes in 240 pathways, including metabolic pathways, pancreatic secretion, protein digestion and absorption, fat digestion and absorption and glycerolipid metabolism. To verify the accuracy of the DGE results, quantitative reverse transcription PCR (qRT-PCR) was performed, focusing on key genes in several related pathways, and the results were highly consistent with the DGE results. Our findings indicated significant differences in cuticular protein genes, ecdysone biosynthesis genes and ecdysone-related nuclear receptors genes, but no significant difference in juvenile hormone and chitin biosynthesis genes was detected. Our research findings lay the foundation for further research on the formation mechanism of the nm2 mutant. PMID:28727825

  5. Mechanisms of larval midgut damage following exposure to phoxim and repair of phoxim-induced damage by cerium in Bombyx mori.

    Science.gov (United States)

    Yu, Xiaohong; Sun, Qingqing; Li, Bing; Xie, Yi; Zhao, Xiaoyang; Hong, Jie; Sheng, Lei; Sang, Xuezi; Gui, Suxin; Wang, Ling; Shen, Weide; Hong, Fashui

    2015-04-01

    Bombyx mori is an important economic animal for silk production. However, it is liable to be infected by organophosphorus pesticide that can contaminate its food and growing environment. It has been known that organophosphorus pesticide including phoxim exposure may damage the digestive systems, produce oxidative stress and neurotoxicity in silkworm B. mori, whereas cerium treatment has been demonstrated to relieve phoxim-induced toxicity in B. mori. However, very little is known about the molecular mechanisms of midgut injury due to phoxim exposure and B. mori protection after cerium pretreatment. The aim of this study was to evaluate the midgut damage and its molecular mechanisms, and the protective role of cerium in B. mori following exposure to phoxim. The results showed that phoxim exposure led to severe midgut damages and oxidative stress; whereas cerium relieved midgut damage and oxidative stress caused by phoxim in B. mori. Furthermore, digital gene expression suggested that phoxim exposure led to significant up-regulation of 94 genes and down-regulation of 52 genes. Of these genes, 52 genes were related with digestion and absorption, specifically, the significant alterations of esterase, lysozyme, amylase 48, and lipase expressions. Cerium pretreatment resulted in up-regulation of 116 genes, and down-regulation of 29 genes, importantly, esterase 48, lipase, lysozyme, and α-amylase were up-regulated. Treatment with Phoxim + CeCl3 resulted in 66 genes up-regulation and 39 genes down-regulation; specifically, levels of esterase 48, lipase, lysozyme, and α-amylase expression in the midgut of silkworms were significantly increased. Therefore, esterase 48, lipase, lysozyme, and α-amylase may be potential biomarkers of midgut toxicity caused by phoxim exposure. These findings may expand the application of rare earths in sericulture. Copyright © 2013 Wiley Periodicals, Inc.

  6. The Structural Variation Is Associated with the Embryonic Lethality of a Novel Red Egg Mutant Fuyin-lre of Silkworm, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Anli Chen

    Full Text Available Bombyx mori presents several types of egg color mutations, all of which have been extensively discussed in sericulture. While the red egg mutation has been previously observed, lethal red-egg mutants have not been reported. In the present work, the red egg mutant Fuyin-lre (Fuyin-lethal red egg was discovered from the Fuyin germplasm resource of B. mori. This mutant features red-colored eggs and embryonic lethality. Genetic analysis showed that Fuyin-lre follows recessive inheritance, with the red egg gene re governing the egg color, and the embryonic lethality of Fuyin-lre may be caused by mutations of other genes closely linked to re. Digital gene expression (DGE was employed to compare the transcription profiles of Fuyin and Fuyin-lre eggs after 24 and 48 h of incubation. A total of 48 differentially expressed genes followed the same expression patterns in both groups at both time points (FDR < 0.01 and log 2 Ratio ≥ 1. Further analyses indicated that 8 out of the 48 genes (including re were closely linked to re. These 8 genes were highly expressed in wild-type Fuyin and the red egg mutant re but showed nearly absent expression in Fuyin-lre. Sequencing of the re gene confirmed that the re gene itself does not induce embryonic lethality, and structure analysis showed that the structural variation of the region where the 8 genes were located may be associated with the embryonic lethality of Fuyin-lre. The present work provides a good foundation for future studies on the mechanism of embryonic lethality and embryonic development in Fuyin-lre.

  7. Produção de casulos de Bombyx mori L. alimentados com dietas artificiais e folhas “in natura” de Morus aba L. Production of cocoon of Bombyx mori L. fed on leaves of Morus aba L. and on artificial diets

    Directory of Open Access Journals (Sweden)

    Adriana Evangelista Rodrigues

    2001-05-01

    Full Text Available A utilização de dietas artificiais no Japão já é uma realidade, e no Brasil iniciam-se os estudos buscando encontrar um balanceamento que satisfaça as necessidades das larvas do bicho-da-seda, Bombyx mori (Lepidoptera. Desta forma, foram testados dois cultivares de amoreira como ingredientes das dietas, avaliando-se a qualidade do alimento, através do peso médio dos casulos e teor líquido de seda. As larvas foram separadas em parcelas, no início do 1º ínstar, onde já começaram a receber as dietas artificiais e as folhas “in natura”. Quando as larvas receberam folhas “in natura”, ao longo dos ínstares, apresentaram valores superiores para peso dos casulos. Por outro lado, quando receberam dietas artificiais, nos dois ínstares iniciais, não apresentaram diferença significativa para teor líquido de seda, quando comparadas com folhas “in natura”. Por fim, observou-se que a variedade Miura é melhor tanto como folha “in natura”, quanto como dieta artificial para peso de casulos.The utilization of artificial diets in Japan is fact and in Brazil the work is beginning to search meeting a balance form to satisfy the silkworm necessity, Bombyx mori (Lepidoptera. In such case, was tested two cultivars of mulberry in diets, to evaluate the quality of food trough mean cocoon weight and liquid silk. The larvae were separated in portion in the first instar begin and were to feed with diets and leaves “in nature”. When the silkworms were fed with leaves during five instars, showed cocoon weight high. When were fed diets in two inicial instars, showed that the silk liquid was good. The Miura variety is better in diets and leaves “in nature” for coccon weight.

  8. Progeny of Palmistichus elaeisis Delvare and LaSalle (Hymenoptera: Eulophidae) parasitising pupae of Bombyx mori L. (Lepidoptera: Bombycidae) of different ages; Progenie de Palmistichus elaeisis Delvare e LaSalle (Hymenoptera:Eulophidae) parasitando pupas de Bombyx mori L. (Lepidoptera:Bombycidae) de diferentes idades

    Energy Technology Data Exchange (ETDEWEB)

    Pereira, Fabricio F.; Favero, Kellen; Grance, Elizangela L.V. [Universidade Federal da Grande Dourados, MS (Brazil). Fac. de Ciencias Biologicas e Ambientais], e-mail: fabriciofagundes@ufgd.edu.br, e-mail: kellenfavero@yahoo.com.br, e-mail: eli_vargasgrance@yahoo.com.br; Zanuncio, Jose C. [Universidade Federal de Vicosa (UFV), MG (Brazil). Dept. de Biologia Animal], e-mail: zanuncio@ufv.br; Serrao, Jose E. [Universidade Federal de Vicosa (UFV), MG (Brazil), Dept. de Biologia Geral], e-mail: jeserrao@ufv.br; Oliveira, Harley N. [Embrapa Agropecuaria Oeste, Dourados, MS (Brazil)], e-mail: harley@cpao.embrapa.br

    2009-09-15

    Palmistichus elaeisis Delvare and LaSalle is a natural pupal parasitoid of eucalyptus defoliator lepidopterans and is considered a promising biocontrol agent. However, the development of efficient rearing techniques for this natural enemy are fi rst required before it can be used in biocontrol programs. Bombyx mori L. pupae are potential alternative hosts for this parasitoid mass rearing, and they are easy to rear. Therefore, we investigated the most suitable host age and the effects of parasitoid age on progeny production of P. elaeisis. B. mori pupae, 24 h-, 48 h-, 72 h- or 96 h-old were exposed to P. elaeisis females of similar age. The duration of the life cycle (egg-adult) of P. elaeisis was not affected by the age of the parasitizing female; however, the host age affected parasitoid development. The best parasitization was obtained for 72h- to 96h-old parasitoid females when offered to 48h- to 72h-old host pupae, allowing the synchronized rearing of a large number of P. elaeisis offspring. (author)

  9. High-titer preparation of Bombyx mori nucleopolyhedrovirus (BmNPV displaying recombinant protein in silkworm larvae by size exclusion chromatography and its characterization

    Directory of Open Access Journals (Sweden)

    Tanaka Shigeyasu

    2009-06-01

    Full Text Available Abstract Background Budded baculoviruses are utilized for vaccine, the production of antibody and functional analysis of transmembrane proteins. In this study, we tried to produce and purify the recombinant Bombyx mori nucleopolyhedrovirus (rBmNPV-hPRR that displayed human (prorenin receptor (hPRR connected with FLAG peptide sequence on its own surface. These particles were used for further binding analysis of hPRR to human prorenin. The rBmNPV-hPRR was produced in silkworm larvae and purified from its hemolymph using size exclusion chromatography (SEC. Results A rapid method of BmNPV titer determination in hemolymph was performed using quantitative real-time PCR (Q-PCR. A correlation coefficient of BmNPV determination between end-point dilution and Q-PCR methods was found to be 0.99. rBmNPV-hPRR bacmid-injected silkworm larvae produced recombinant baculovirus of 1.31 × 108 plaque forming unit (pfu in hemolymph, which was 2.8 × 104 times higher than transfection solution in Bm5 cells. Its purification yield by Sephacryl S-1000 SF column chromatography was 264 fold from larval hemolymph at 4 days post-injection (p.i., but 35 or 39 fold at 4.5 or 5 days p.i., respectively. Protein patterns of rBmNPV-hPRR purified at 4 and 5 days were the same and ratio of envelope proteins (76, 45 and 35 kDa to VP39, one of nucleocapsid proteins, increased at 5 days p.i. hPRR was detected in only purified rBmNPV-hPRR at 5 days p.i.. Conclusion The successful purification of rBmNPV-hPRR indicates that baculovirus production using silkworm larvae and its purification from hemolymph by Sephacryl S-1000 SF column chromatography can provide an economical approach in obtaining the purified BmNPV stocks with high titer for large-scale production of hPRR. Also, it can be utilized for further binding analysis and screening of inhibitors of hPRR.

  10. Tachykinin-Related Peptides Share a G Protein-Coupled Receptor with Ion Transport Peptide-Like in the Silkworm Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Chiaki Nagai-Okatani

    Full Text Available Recently, we identified an orphan Bombyx mori neuropeptide G protein-coupled receptor (BNGR-A24 as an ion transport peptide-like (ITPL receptor. BNGR-A24 belongs to the same clade as BNGR-A32 and -A33, which were recently identified as natalisin receptors. Since these three BNGRs share high similarities with known receptors for tachykinin-related peptides (TRPs, we examined whether these BNGRs can function as physiological receptors for five endogenous B. mori TRPs (TK-1-5. In a heterologous expression system, BNGR-A24 acted as a receptor for all five TRPs. In contrast, BNGR-A32 responded only to TK-5, and BNGR-A33 did not respond to any of the TRPs. These findings are consistent with recent studies on the ligand preferences for B. mori natalisins. Furthermore, we evaluated whether the binding of ITPL and TRPs to BNGR-A24 is competitive by using a Ca2+ imaging assay. Concomitant addition of a TRP receptor antagonist, spantide I, reduced the responses of BNGR-A24 not only to TK-4 but also to ITPL. The results of a binding assay using fluorescent-labeled BNGR-A24 and ligands demonstrated that the binding of ITPL to BNGR-A24 was inhibited by TK-4 as well as by spantide I, and vice versa. In addition, the ITPL-induced increase in cGMP levels of BNGR-A24-expressing BmN cells was suppressed by the addition of excess TK-4 or spantide I. The intracellular levels of cAMP and cGMP, as second messenger candidates of the TRP signaling, were not altered by the five TRPs, suggesting that these peptides act via different signaling pathways from cAMP and cGMP signaling at least in BmN cells. Taken together, the present findings suggest that ITPL and TRPs are endogenous orthosteric ligands of BNGR-A24 that may activate discrete signaling pathways. This receptor, which shares orthosteric ligands, may constitute an important model for studying ligand-biased signaling.

  11. Tachykinin-Related Peptides Share a G Protein-Coupled Receptor with Ion Transport Peptide-Like in the Silkworm Bombyx mori.

    Science.gov (United States)

    Nagai-Okatani, Chiaki; Nagasawa, Hiromichi; Nagata, Shinji

    2016-01-01

    Recently, we identified an orphan Bombyx mori neuropeptide G protein-coupled receptor (BNGR)-A24 as an ion transport peptide-like (ITPL) receptor. BNGR-A24 belongs to the same clade as BNGR-A32 and -A33, which were recently identified as natalisin receptors. Since these three BNGRs share high similarities with known receptors for tachykinin-related peptides (TRPs), we examined whether these BNGRs can function as physiological receptors for five endogenous B. mori TRPs (TK-1-5). In a heterologous expression system, BNGR-A24 acted as a receptor for all five TRPs. In contrast, BNGR-A32 responded only to TK-5, and BNGR-A33 did not respond to any of the TRPs. These findings are consistent with recent studies on the ligand preferences for B. mori natalisins. Furthermore, we evaluated whether the binding of ITPL and TRPs to BNGR-A24 is competitive by using a Ca2+ imaging assay. Concomitant addition of a TRP receptor antagonist, spantide I, reduced the responses of BNGR-A24 not only to TK-4 but also to ITPL. The results of a binding assay using fluorescent-labeled BNGR-A24 and ligands demonstrated that the binding of ITPL to BNGR-A24 was inhibited by TK-4 as well as by spantide I, and vice versa. In addition, the ITPL-induced increase in cGMP levels of BNGR-A24-expressing BmN cells was suppressed by the addition of excess TK-4 or spantide I. The intracellular levels of cAMP and cGMP, as second messenger candidates of the TRP signaling, were not altered by the five TRPs, suggesting that these peptides act via different signaling pathways from cAMP and cGMP signaling at least in BmN cells. Taken together, the present findings suggest that ITPL and TRPs are endogenous orthosteric ligands of BNGR-A24 that may activate discrete signaling pathways. This receptor, which shares orthosteric ligands, may constitute an important model for studying ligand-biased signaling.

  12. Molecular diagnosis for the silk worm Bombyx Mori L. Viral and bacterial diseases in the irradiated and non-irradiated individuals

    International Nuclear Information System (INIS)

    Abulyazid, I.; Elshafei, A.; El-Said, E.; Mousa, S.; Taha, R.

    2007-01-01

    Genetic maps for the Bombyx Mori infectious flacherrie virus (BmlFV) causing flacherrie (Fl) disease and nuclear polyhedrosis virus (BmNPV) causing grasserie (Gr) disease were built up in an attempt to diagnose diseases early in young larval stages. For the non-irradiated and irradiated viral RNA of IFV, no amplification was obtained by using RT-PCR and RAPD-PCR techniques. HcoRI, EcoRV, BamHI, Hind III and BamHI restriction enzymes were used to digest the non-irradiated and irradiated viral DNA of BmNPV. It was found that, the two viral DNA samples were genetically different; the similarity indexes were 0.14, 0, 0, 0.18 and 0.15, respectively. At the biochemical level, native protein electrophoresis showed 4 and 3 new proteins in non-irradiated and irradiated Fl diseased larvae, respectively, while Gr diseased larvae showed 1 and 3 new protein types. The similarity index (S.I) between all the tested samples was not exceeded 44%. For lipoprotein pattern, 2 and 3 new lipoprotein types were appeared due to Fl disease in the non-irradiated and irradiated haemolymph samples, respectively, while Gr disease showed 3 new lipoproteins in the non-irradiated samples only. The highest S.I recorded was 56%. Glycoprotein pattern revealed 3 and 5 new glycoprotein types appeared due to Fl disease while Gr disease showed 4 and 6 new types in the non-irradiated and irradiated samples, respectively. The highest S.I was 77%. Fractionated protein with SDS revealed 2 common bands shared between the tested samples with R f values 0.28 and 0.71. Fl disease increased the number of protein bands with the appearance of 5 and 4 new proteins types. Gr disease reduced the total number of proteins with the appearance of 2 and 3 new types. The highest S.I was 59%. Both diseases and irradiation may be mutagenic through the epigenetic level in silkworm larvae leading to death. Thus, the results of the biochemical and genetic characterization of IFV and BmNPV enable us to conclude that the

  13. Aminopeptidase N isoforms from the midgut of Bombyx mori and Plutella xylostella -- their classification and the factors that determine their binding specificity to Bacillus thuringiensis Cry1A toxin.

    Science.gov (United States)

    Nakanishi, Kazuko; Yaoi, Katsuro; Nagino, Yasushi; Hara, Hirotaka; Kitami, Madoka; Atsumi, Shogo; Miura, Nami; Sato, Ryoichi

    2002-05-22

    Novel aminopeptidase N (APN) isoform cDNAs, BmAPN3 and PxAPN3, from the midguts of Bombyx mori and Plutella xylostella, respectively, were cloned, and a total of eight APN isoforms cloned from B. mori and P. xylostella were classified into four classes. Bacillus thuringiensis Cry1Aa and Cry1Ab toxins were found to bind to specific APN isoforms from the midguts of B. mori and P. xylostella, and binding occurred with fragments that corresponded to the BmAPN1 Cry1Aa toxin-binding region of each APN isoform. The results suggest that APN isoforms have a common toxin-binding region, and that the apparent specificity of Cry1Aa toxin binding to each intact APN isoform seen in SDS-PAGE is determined by factors such as expression level in conjunction with differences in binding affinity.

  14. Leaf Surface Scanning Electron Microscopy of 16 Mulberry Genotypes (Morus spp. with Respect to their Feeding Value in Silkworm (Bombyx mori L. Rearing Microscopía Electrónica de Barrido de la Superficie Foliar de 16 Genotipos de Morus spp. en Relación a su Valor Alimenticio para Crianza del Gusano de la Seda (Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    B.K Singhal

    2010-06-01

    Full Text Available Mulberry (Morus spp. is the only silkworm (Bombyx mori L. food plant. In Indian sub tropics, S-146 is the only popular and ruling mulberry genotype for silkworm rearing. As a result, mulberry leaf availability is always the limiting factor, and therefore, sub tropics are contributing less than 1% of the country’s total silk production compared with more than 60% under tropical conditions. Besides climatic conditions, this is due to a very limited number of mulberry genotypes available in this region for silkworm rearing. However, in the mean time, 15 mulberry genotypes viz. ‘Tr-10’,‘Chinese White’,‘K-2’,‘Sujanpur Local’,‘BC2-59’,‘S-1635’,‘C-1730’,‘Mandalaya’,‘S-30’‘(Vishala,‘RFS-175’,‘Anantha’,‘C-2016’,‘C-2017’,‘S-41’ and‘V-1’ were also introduced in the sub tropics, but remained unexplored. In sericulture, leaf surface is also an important parameter for, both, the silkworm’s acceptability of not having any feeding impediment and the mulberry improvement programs. The objective of this study was to explore the possibilities of using these 16 mulberry genotypes for their leaf surface characteristics by scanning electron microscopy and using them for sericulture. Based on leaf yield, stomatal size, stomatal number per unit of area and trichomes and idioblasts length, these genotypes were grouped into different categories. The mulberry genotype ‘Mandalaya’, in addition to the ruling genotype ‘S-146’ excelled because of their higher leaf yield and desired leaf surface characteristics. Furthermore, the genotypes ‘K-2’, ‘S-41’ and ‘Sujanpur Local’ are also suggested to develop high yield mulberry genotypes in the Indian sub tropics.La morera (Morus spp. es la única planta de alimento para el gusano de la seda (Bombyx mori L.. En los sub-trópicos de la India, ‘S-146’ es el único genotipo popular y predominante de morera para criarlo. Como resultado, la

  15. iTRAQ-based quantitative proteomics analysis of molecular mechanisms associated with Bombyx mori (Lepidoptera) larval midgut response to BmNPV in susceptible and near-isogenic strains.

    Science.gov (United States)

    Yu, Haizhong; Wang, Xueyang; Xu, Jiaping; Ma, Yan; Zhang, Shangzhi; Yu, Dong; Fei, Dongqiong; Muhammad, Azharuddin

    2017-08-08

    Bombyx mori nucleopolyhedrovirus (BmNPV) has been identified as a major pathogen responsible for severe economic loss. Most silkworm strains are susceptible to BmNPV, with only a few highly resistant strains thus far identified. Here we investigated the molecular basis of silkworm resistance to BmNPV using susceptible (the recurrent parent P50) and resistant (near-isogenic line BC9) strains and a combination of iTRAQ-based quantitative proteomics, reverse-transcription quantitative PCR and Western blotting. By comparing the proteomes of infected and non-infected P50 and BC9 silkworms, we identified 793 differentially expressed proteins (DEPs). By gene ontology and KEGG enrichment analyses, we found that these DEPs are preferentially involved in metabolism, catalytic activity, amino sugar and nucleotide sugar metabolism and carbon metabolism. 114 (14.38%) DEPs were associated with the cytoskeleton, immune response, apoptosis, ubiquitination, translation, ion transport, endocytosis and endopeptidase activity. After removing the genetic background and individual immune stress response proteins, we identified 84 DEPs were found that are potentially involved in resistance to BmNPV. Further studies showed that a serine protease was down-regulated in P50 and up-regulated in BC9 after BmNPV infection. Taken together, these results provide insights into the molecular mechanism of silkworm response to BmNPV. Bombyx mori nucleopolyhedrovirus (BmNPV) is highly pathogenic, causing serious losses in sericulture every year. However, the molecular mechanisms of BmNPV infection and host defence remain unclear. Here we combined quantitative proteomic, bioinformatics, RT-qPCR and Western blotting analyses and found that BmNPV invasion causes complex protein alterations in the larval midgut, and that these changes are related to cytoskeleton, immune response, apoptosis, ubiquitination, translation, ion transport, endocytosis and endopeptidase activity. Five important differentially

  16. The domain II loops of Bacillus thuringiensis Cry1Aa form an overlapping interaction site for two Bombyx mori larvae functional receptors, ABC transporter C2 and cadherin-like receptor.

    Science.gov (United States)

    Adegawa, Satomi; Nakama, Yui; Endo, Haruka; Shinkawa, Naoki; Kikuta, Shingo; Sato, Ryoichi

    2017-02-01

    Information about the receptor-interaction region of Cry toxins, insecticidal proteins produced by Bacillus thuringiensis, is needed to elucidate the mode of action of Cry toxins and improve their toxicity through protein engineering. We analyzed the interaction sites on Cry1Aa with ABC transporter C2 (ABCC2), one of the most important Cry1A toxin receptors. A competitive binding assay revealed that the Bombyx mori ABCC2 (BmABCC2) Cry1A binding site was the same as the BtR175 binding site, suggesting that the loop region of Cry1Aa domain II is a binding site. Next, we constructed several domain II loop mutant toxins and tested their binding affinity in an SPR analysis, and also performed a cell swelling assay to evaluate receptor-mediated cytotoxicity. Our results indicate that the loop regions required for BtR175 and BmABCC2 binding and the regions important for cytotoxicity partially overlap. Our results also suggest that receptor binding is necessary but not sufficient for cytotoxicity. This is the first report showing the region of interaction between ABCC2 and Cry1Aa and the cytotoxicity-relevant properties of the Cry1Aa domain II loop region. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Microscopic structural analysis of fractured silk fibers from Bombyx mori and Samia cynthia ricini using 13C CP/MAS NMR with a 1 mm microcoil MAS NMR probehead

    KAUST Repository

    Yamauchi, Kazuo

    2010-07-01

    Conformational changes have been studied in silk fibers from the domestic silkworm Bombyx mori and a wild silkworm Samia cynthia ricini as a result of fractured by stretching. About 300 samples consisting of only the fractured regions of [1-13C]Ala or [1-13C]Gly labeled silk fibers were collected and observed by 13C CP/MAS NMR spectra. The total amount of these fractured fibers is only about 1 mg and therefore we used a home-built 1 mm microcoil MAS NMR probehead. A very small increase in the fraction of random coil was noted for the alanine regions of both silk fibroins and for the glycine region of B. mori silk fibroin. However, there is no difference in the spectra before and after fractured for the glycine region of S. c. ricini silk fibroin. Thus, the influence of fracture occurs exclusively at the Ala region for S. c. ricini. The relationship between sequence, fracture and structure is discussed. © 2010 Elsevier Inc. All rights reserved.

  18. Molecular tracing of white muscardine in the silkworm, Bombyx mori (Linn.) II. Silkworm white muscardine is not caused by artificial release or natural epizootic of Beauveria bassiana in China.

    Science.gov (United States)

    Chen, Xue; Huang, Cui; He, Lingmin; Zhang, Shengli; Li, Zengzhi

    2015-02-01

    The fungal pathogen Beauveria bassiana causes serious economic losses in sericulture. Its origin is usually attributed to the release of B. bassiana insecticides against pine caterpillars (Dendrolimus punctuatus). In the present study, 488 B. bassiana isolates obtained from silkworm (Bombyx mori) collected from 13 Chinese provinces, and 327 B. bassiana isolates obtained from D. punctatus collected from 9 provinces, were analyzed for population genetic structure using the ISSR technique based on genetic distance. A UPGMA dendrogram clustered them into three independent clades: two B. mori clades and one D. punctatus clade. A 3-D principal component analysis further divided them into two completely independent host groups, revealing high host-specificity. This suggested that white muscardine occurring in B. mori populations throughout southern China was not caused by any B. bassiana strain either naturally prevailing in D. punctatus populations or by any strain artificially released as a fungal insecticide against D. punctatus. We further investigated the genetic differentiation coefficient Gst and gene flow between B. mori-pathogenic and D. punctatus-pathogenic B. bassiana isolates from across China and from five provinces inhabited by both B. mori and D. punctatus. The Gst value across China was computed as 0.410, while the values of the five provinces ranged from 0.508 to 0.689; all above 0.25, which is the threshold for significant genetic differentiation. This suggests that B. bassiana strains isolated from the two different hosts maintained their respective heredity without a convergent homogenization trend, and reduces the possibility that the host range of the caterpillar isolates could expand and enhance their virulence in B. mori. These findings indicate that the use of B. bassiana does not threaten the safety of sericulture. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. Single amino acid insertions in extracellular loop 2 of Bombyx mori ABCC2 disrupt its receptor function for Bacillus thuringiensis Cry1Ab and Cry1Ac but not Cry1Aa toxins.

    Science.gov (United States)

    Tanaka, Shiho; Miyamoto, Kazuhisa; Noda, Hiroaki; Endo, Haruka; Kikuta, Shingo; Sato, Ryoichi

    2016-04-01

    In a previous report, seven Cry1Ab-resistant strains were identified in the silkworm, Bombyx mori; these strains were shown to have a tyrosine insertion at position 234 in extracellular loop 2 of the ABC transporter C2 (BmABCC2). This insertion was confirmed to destroy the receptor function of BmABCC2 and confer the strains resistance against Cry1Ab and Cry1Ac. However, these strains were susceptible to Cry1Aa. In this report, we examined the mechanisms of the loss of receptor function of the transporter by expressing mutations in Sf9 cells. After replacement of one or two of the five amino acid residues in loop 2 of the susceptible BmABCC2 gene [BmABCC2_S] with alanine, cells still showed susceptibility, retaining the receptor function. Five mutants with single amino acid insertions at position 234 in BmABCC2 were also generated, resulting in loop 2 having six amino acids, which corresponds to replacing the tyrosine insertion in the resistant BmABCC2 gene [BmABCC2_R(+(234)Y)] with another amino acid. All five mutants exhibited loss of function against Cry1Ab and Cry1Ac. These results suggest that the amino acid sequence in loop 2 is less important than the loop size (five vs. six amino acids) or loop structure for Cry1Ab and Cry1Ac activity. Several domain-swapped mutant toxins were then generated among Cry1Aa, Cry1Ab, and Cry1Ac, which are composed of three domains. Swapped mutants containing domain II of Cry1Ab or Cry1Ac did not kill Sf9 cells expressing BmABCC2_R(+(234)Y), suggesting that domain II of the Cry toxin is related to the interaction with the receptor function of BmABCC2. This also suggests that different reactions against Bt-toxins in some B. mori strains, that is, Cry1Ab resistance or Cry1Aa susceptibility, are attributable to structural differences in domain II of Cry1A toxins. Copyright © 2016. Published by Elsevier Inc.

  20. Molecular phylogeny of the domesticated silkworm, Bombyx mori ...

    Indian Academy of Sciences (India)

    The sequences of the 16 silkworm strains were analysed with DNASTAR software and a phylogenic tree was constructed using PHYLIP software. ... 221 116, China; Shanxi Institute of Education, Xi'an 710 061, China; Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100 081, China ...

  1. Subcelluar localization of orf126 of Bombyx mori nucleopolyhedrovirus

    African Journals Online (AJOL)

    Jane

    2011-08-15

    Aug 15, 2011 ... The plasmids containing the different subtypes of orf126-egfp fusion gene were constructed to study the ... Localization of the SX126 and GD126 EGFP-fusion proteins in BmN cells with or without virus ... with BmBacJS13-ph at 12 h p.t., the nuclear DNAs of the cells were stained with Hoechst and were ...

  2. (Bombyx mori L.) using two-dimensional polyacrylami

    Indian Academy of Sciences (India)

    Unknown

    Fountoulakis M, Schuller E, Hardmeier R, Berndt P and Lubec. G 1999 Rat brain proteins: Two-dimensional protein data- base and variation in the expression level; Electrophoresis 20. 3527–3579. Hiroshi Fujii, Junji Tochinara, Yutaka Kawaguchi and Sakagu- chi B 1988 Existence of carotenoids binding protein in larval.

  3. Molecular phylogeny of the domesticated silkworm, Bombyx mori ...

    Indian Academy of Sciences (India)

    The Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212 018, Jiangsu Province, China; College of Life Sciences, Xuzhou Normal University, Xuzhou 221 116, China; Shanxi Institute of Education, Xi'an 710 061, China; Biotechnology Research Institute, Chinese Academy of Agricultural ...

  4. Molecular phylogeny of the domesticated silkworm, Bombyx mori ...

    Indian Academy of Sciences (India)

    Unknown

    2The Sericultural Research Institute, Chinese Academy of Agricultural Sciences,. Zhenjiang 212 018, Jiangsu Province, China. 3College of Life Sciences, Xuzhou Normal University, Xuzhou 221 116, China. 4Present address: Shanxi Institute of Education, Xi'an 710 061, China. Abstract. Pupae from the Chinese wild ...

  5. Study of 31 economically important traits in 20 silkworm Bombyx ...

    African Journals Online (AJOL)

    AJL

    2012-05-03

    May 3, 2012 ... Silkworm produces silk as high quality fiber and silk fabrics are highly attractive. There are different lines and strains for silk production. Production of cocoon and raw silk are affected by several factors, such as genetic potential of commercial varieties, quality of silkworm eggs, pests and diseases incidence ...

  6. Transmission Electron Microscopy of Bombyx Mori Silk Fibers

    Science.gov (United States)

    Shen, Y.; Martin, D. C.

    1997-03-01

    The microstructure of B. Mori silk fibers before and after degumming was examined by TEM, selected area electron diffraction (SAED), WAXS and low voltage SEM. SEM micrographs of the neat cocoon revealed a network of pairs of twisting filaments. After degumming, there were only individual filaments showing a surface texture consistent with an oriented fibrillar structure in the fiber interior. WAXS patterns confirmed the oriented beta-sheet crystal structure common to silkworm and spider silks. Low dose SAED results were fully consistent with the WAXS data, and revealed that the crystallographic texture did not vary significantly across the fiber diameter. TEM observations of microtomed fiber cross sections indicated a somewhat irregular shape, and also revealed a 0.5-2 micron sericin coating which was removed by the degumming process. TEM observations of the degummed silk fiber showed banded features with a characteristic spacing of nominally 600 nm along the fiber axis. These bands were oriented in a roughly parabolic or V-shape pointing along one axis within a given fiber. We hypothesize that this orientation is induced by the extrusion during the spinning process. Equatorial DF images revealed that axial and lateral sizes of the β-sheet crystallites in silk fibroin ranged from 20 to 170 nm and from 1 to 24 nm, respectively. Crazes developed in the degummed silk fiber parallel to the fiber direction. The formation of these crazes suggests that there are significant lateral interactions between fibrils in silk fibers.

  7. Comparative analysis of two acetylcholinesterase genes of Bombyx ...

    African Journals Online (AJOL)

    To compare the sequences and tissue expressions of the two aces between the two species, cDNAs encoding two ace genes were cloned and designated as Bmm-ace1 and Bmm-ace2 from the larvae of the B. mandarina. The amino acid sequence of Bmm-ace1 shared 99.71% homology with its homolog, Bm-ace1, in B.

  8. Genetic diversity and classification of 51 strains of silkworm Bombyx ...

    African Journals Online (AJOL)

    The aim of this experiment was to study and classify all 51 pure lines of Iran silkworm germplasm based on larval traits and identification of pure lines relationships. The average linkage between two groups is considered as the average of distance between all pairs of cases with one number from each group. Hierarchical ...

  9. Effect of ultrasound on protein metabolism in the silkworm, Bombyx ...

    African Journals Online (AJOL)

    Ultrasound was found to promote the accumulation of proteins, which include silk proteins as well, while retarding proteolysis and turnover of proteins towards the release of amino acids, keto-acids etc. Changes in the levels of these biochemical constituents are correlated with the events of histogenesis and histolysis ...

  10. Genome engineering and parthenocloning in the silkworm, Bombyx mori

    Czech Academy of Sciences Publication Activity Database

    Zabelina, V.; Klymenko, V.; Tamura, T.; Doroshenko, K.; Liang, H.; Sezutsu, H.; Sehnal, František

    2015-01-01

    Roč. 40, č. 3 (2015), s. 645-655 ISSN 0250-5991 R&D Projects: GA MŠk(CZ) EE2.3.30.0032; GA ČR GAP502/10/2382 Institutional support: RVO:60077344 Keywords : genomic cloning * genetic engineering * insect biotechnology Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.419, year: 2015 http://www.ias.ac.in/jbiosci/sep2015/645.pdf

  11. Genome engineering and parthenocloning in the silkworm, Bombyx ...

    Indian Academy of Sciences (India)

    Several methods of gene targeting are being developed to minimize position effect on transgene expression and facilitate cloning. Parthenocloning can be exploited to conserve genetic traits and improve selection and amplification of clones containing genes of interest. Some silkworm clones have been bred for decades ...

  12. Genetic diversity and classification of 51 strains of silkworm Bombyx ...

    African Journals Online (AJOL)

    GREGORY

    2010-09-27

    Sep 27, 2010 ... silkworm (Gamo 1983; Chatterjee and Data, 1992). As sericultural regions of the world have different climatic conditions, physiological diversification has also been influenced by agro-climatic factors. Thus, given geographic isolation and limited cultural exchange, some strains may have acquired similar ...

  13. Cloning and homologic analysis of Tpn I gene in silkworm Bombyx ...

    African Journals Online (AJOL)

    GREGO

    2007-03-19

    Mar 19, 2007 ... calcium conditions, tropomyosin locks the actin site of myosin binding. When the Calcium level increases in the sarcoplasm, Troponin C is able to ... encing was performed using an automatic sequencer: CEQ8000. (Beckman USA). Software and database used for bioinformatic analysis. The primer used to ...

  14. Complete resequencing of 40 genomes reveals domestication events and genes in silkworm (Bombyx)

    DEFF Research Database (Denmark)

    Xia, Qingyou; Guo, Yiran; Zhang, Ze

    2009-01-01

    genes that may have been important during domestication, some of which have enriched expression in the silk gland, midgut, and testis. These data add to our understanding of the domestication processes and may have applications in devising pest control strategies and advancing the use of silkworms...

  15. Targeted mutagenesis in the silkworm Bombyx mori using zinc finger nuclease mRNA injection

    Czech Academy of Sciences Publication Activity Database

    Takasu, Yoko; Kobayashi, I.; Beumer, K.; Uchino, K.; Sezutsu, H.; Sajwan, S.; Carroll, D.; Tamura, T.; Žurovec, Michal

    2010-01-01

    Roč. 40, č. 10 (2010), s. 759-765 ISSN 0965-1748 R&D Projects: GA AV ČR IAA500070601; GA ČR GAP305/10/2406; GA MŠk(CZ) LC06077 Institutional research plan: CEZ:AV0Z50070508 Keywords : gene targeting * nonhomologous and joining * Lepidoptera Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.018, year: 2010

  16. The wings of Bombyx mori develop from larval discs exhibiting an ...

    Indian Academy of Sciences (India)

    Unknown

    field and French 1999). We present here the expression pattern of Nubbin (Nub), Wingless (Wg) and Distal-less. (Dll) in B. mori wing discs using antibodies generated against the corresponding proteins from Drosophila or the butterfly P. coenia. The expression of Wg and Dll as overlapping domains in the distal region in B.

  17. Proteomic-based insight into Malpighian tubules of silkworm Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Xiao-wu Zhong

    Full Text Available Malpighian tubules (MTs are highly specific organs of arthropods (Insecta, Myriapoda and Arachnida for excretion and osmoregulation. In order to highlight the important genes and pathways involved in multi-functions of MTs, we performed a systematic proteomic analysis of silkworm MTs in the present work. Totally, 1,367 proteins were identified by one-dimensional gel electrophoresis coupled with liquid chromatography-tandem mass spectrometry, and as well as by Trans Proteomic Pipeline (TPP and Absolute protein expression (APEX analyses. Forty-one proteins were further identified by two-dimensional gel electrophoresis. Some proteins were revealed to be significantly associated with various metabolic processes, organic solute transport, detoxification and innate immunity. Our results might lay a good foundation for future functional studies of MTs in silkworm and other lepidoptera.

  18. Assessment of bioactivity of cassia fistula using bombyx mori lethality assay

    International Nuclear Information System (INIS)

    Ashfaq, M.

    2009-01-01

    In recent times, focus on plant research has increased all over the world and a large body of evidence is being collected to evaluate immense potential of medicinal plants used in various traditional systems. In this regard, the present study was conducted to evaluate the bioactivity of a commonly used medicinal plant Cassia fistula against newly selected Bambyx mori (silkworm) larvae. C. fistula pods were extracted using water, methanol, ethanol, hydro-methanol (1: I) and hydro-ethanol (1: I) and were assayed for their activity against Bambyx mori. Methanol extract of C fistula at concentration of 100 mg/L killed half (LC/sub 50/) of Bamyx mori larvae under study. Bomyx mori LC/sub 50/ for other C. fistula extracts were 400 mg/L for ethanol and hydro-methanol, 800 mg/L for hydro-ethanol and 1600 mg/L for aqueous extract. From the results of the present study it can be concluded that Bambyx mori lethality bioassay can be considered a useful preliminary tool for plant extract toxicity evaluation. The main objectives of the present study was to develop a new and simple assay to evaluate claims from traditional. tribal and advanced medicinal lore to suggest directions for future clinical research and commercial importance that could be carried out by local investigators in developing regions. (author)

  19. Biochemical characterization of maintenance DNA methyltransferase DNMT-1 from silkworm, Bombyx mori.

    Science.gov (United States)

    Mitsudome, Takumi; Mon, Hiroaki; Xu, Jian; Li, Zhiqing; Lee, Jae Man; Patil, Anandrao Ashok; Masuda, Atsushi; Iiyama, Kazuhiro; Morokuma, Daisuke; Kusakabe, Takahiro

    2015-03-01

    DNA methylation is an important epigenetic mechanism involved in gene expression of vertebrates and invertebrates. In general, DNA methylation profile is established by de novo DNA methyltransferases (DNMT-3A, -3B) and maintainance DNA methyltransferase (DNMT-1). DNMT-1 has a strong substrate preference for hemimethylated DNA over the unmethylated one. Because the silkworm genome lacks an apparent homologue of de novo DNMT, it is still unclear that how silkworm chromosome establishes and maintains its DNA methylation profile. As the first step to unravel this enigma, we purified recombinant BmDNMT-1 using baculovirus expression system and characterized its DNA-binding and DNA methylation activity. We found that the BmDNMT-1 preferentially methylates hemimethylated DNA despite binding to both unmethylated and hemimethylated DNA. Interestingly, BmDNMT-1 formed a complex with DNA in the presence or absence of methyl group donor, S-Adenosylmethionine (AdoMet) and the AdoMet-dependent complex formation was facilitated by Zn(2+) and Mn(2+). Our results provide clear evidence that BmDNMT-1 retained the function as maintenance DNMT but its sensitivity to metal ions is different from mammalian DNMT-1. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Expression and Functional Analysis of Storage Protein 2 in the Silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Wei Yu

    2013-01-01

    Full Text Available Storage protein 2 (SP2 not only is an important source of energy for the growth and development of silkworm but also has inhibitory effects on cell apoptosis. Endothelial cell (EC apoptosis is an important contributing factor in the development of atherosclerosis; therefore, study of the antiapoptotic activity of SP2 on ECs provides information related to the treatment of atherosclerosis and other cardiovascular diseases. In this study, the sp2 gene was cloned and expressed in Escherichia coli to produce a 6xHis-tagged fusion protein, which was then used to generate a polyclonal antibody. Western blot results revealed that SP2 levels were higher in the pupal stage and hemolymph of fifth-instar larvae but low in the egg and adult stages. Subcellular localization results showed that SP2 is located mainly on the cell membrane. In addition, a Bac-to-Bac system was used to construct a recombinant baculovirus for SP2 expression. The purified SP2 was then added to a culture medium for human umbilical vein ECs (HUVECs, which were exposed to staurosporine. A cell viability assay demonstrated that SP2 could significantly enhance the viability of HUVEC. Furthermore, both ELISA and flow cytometry results indicated that SP2 has anti-apoptotic effects on staurosporine-induced HUVEC apoptosis.

  1. [Chromatin in diapause of the silkworm Bombyx mori L.: thermal parthenogenesis and normal development].

    Science.gov (United States)

    Klimenko, V V; Khaoiuan', Lian

    2012-01-01

    Having used hematoxylin as a stain, some features of silkworm embryo chromatin in diapause have been studied in normal and parthenogenetic development. With found direct correlation between the number of interphase chromatin grains and the number of chromosomes in the nucleus, we examined cell polyploidization in the embryo at diapause stage. Polyploidization by parthenogenesis is not reducible to endomitotic doubling of the chromosome set because it comprises 6n-nuclei. Explanation of more diverse range of polyploid cells in parthenogenesis needs to consider the fusion of cleavage nuclei that is carried out by the cytoplasmic karyogamic mechanism in the absence of fertilization. For the first time on squash preparations, in diapausing embryo, we have identified primary germ cells (PGC) that are characterized by less compact chromatin, especially in the zygotic form of development, a larger size of the nucleus and cytoplasm, and irregular number and size of nucleoli. Evaluation of PGC ploidy in parthenogenesis by calculation of "loose" chromatin grains in diapause is possible and testifies polyploidization in embryo germ-line. This explains the inevitable admixture of tetraploid eggs in diploid parthenoclone grain and its absence in normal development. Cytological method used has revealed a spiral arrangement of chromatin grains on the inner surface of the nucleus at different levels of ploidy.

  2. The wings of Bombyx mori develop from larval discs exhibiting an ...

    Indian Academy of Sciences (India)

    Unknown

    and meta-thoracic discs continue to grow in size with larval age and do not display any massive cell rearrangement during metamor- phosis (compare c with d, and h with i) during the larval to pupal .... to surgery. The conspicuous absence of wing blades and presence of wound healing marks on one side are evident.

  3. Molecular defect of isovaleryl-CoA dehydrogenase in the skunk mutant of silkworm, Bombyx mori.

    Science.gov (United States)

    Urano, Kei; Daimon, Takaaki; Banno, Yutaka; Mita, Kazuei; Terada, Tohru; Shimizu, Kentaro; Katsuma, Susumu; Shimada, Toru

    2010-11-01

    The isovaleric acid-emanating silkworm mutant skunk (sku) was first studied over 30 years ago because of its unusual odour and prepupal lethality. Here, we report the identification and characterization of the gene responsible for the sku mutant. Because of its specific features and symptoms similar to human isovaleryl-CoA dehydrogenase (IVD) deficiency, also known as isovaleric acidaemia, IVD dysfunction in silkworms was predicted to be responsible for the phenotype of the sku mutant. Linkage analysis revealed that the silkworm IVD gene (BmIVD) was closely linked to the odorous phenotype as expected, and a single amino acid substitution (G376V) was found in BmIVD of the sku mutant. To investigate the effect of the G376V substitution on BmIVD function, wild-type and sku-type recombinants were constructed with a baculovirus expression system and the subsequent enzyme activity of sku-type BmIVD was shown to be significantly reduced compared with that of wild-type BmIVD. Molecular modelling suggested that this reduction in the enzyme activity may be due to negative effects of G376V mutation on FAD-binding or on monomer-monomer interactions. These observations strongly suggest that BmIVD is responsible for the sku locus and that the molecular defect in BmIVD causes the characteristic smell and prepupal lethality of the sku mutant. To our knowledge, this is, aside from humans, the first characterization of IVD deficiency in metazoa. Considering that IVD acts in the third step of leucine degradation and the sku mutant accumulates branched-chain amino acids in haemolymph, this mutant may be useful in the investigation of unique branched-chain amino acid catabolism in insects. © 2010 The Authors Journal compilation © 2010 FEBS.

  4. Glutathione-binding site of a bombyx mori theta-class glutathione transferase.

    Directory of Open Access Journals (Sweden)

    M D Tofazzal Hossain

    Full Text Available The glutathione transferase (GST superfamily plays key roles in the detoxification of various xenobiotics. Here, we report the isolation and characterization of a silkworm protein belonging to a previously reported theta-class GST family. The enzyme (bmGSTT catalyzes the reaction of glutathione with 1-chloro-2,4-dinitrobenzene, 1,2-epoxy-3-(4-nitrophenoxy-propane, and 4-nitrophenethyl bromide. Mutagenesis of highly conserved residues in the catalytic site revealed that Glu66 and Ser67 are important for enzymatic function. These results provide insights into the catalysis of glutathione conjugation in silkworm by bmGSTT and into the metabolism of exogenous chemical agents.

  5. Identification and characterization of Bms3a in Bombyx mori L.

    African Journals Online (AJOL)

    STORAGESEVER

    2008-10-06

    Oct 6, 2008 ... The heredity of silkworm resistance to NPV is relatively complicated because it is controlled both by major dominant genes and multiple micro-effect genes (Chen et al., 2003). Though significant progress has been made in breeding highly resistant strains (Chen et al., 1991, 1996), and some molecular.

  6. Sericin removal from raw Bombyx mori silk scaffolds of high hierarchical order.

    Science.gov (United States)

    Teuschl, Andreas Herbert; van Griensven, Martijn; Redl, Heinz

    2014-05-01

    Silk fibroin has previously been described as a promising candidate for ligament tissue engineering (TE) approaches. For biocompatibility reasons, silkworm silk requires removal of sericin, which can elicit adverse immune responses in the human body. One disadvantage of the required degumming process is the alteration of the silk fiber structural properties, which can hinder textile engineering of high order hierarchical structures. Therefore, the aim of this study was to find a way to remove sericin from a compact and highly ordered raw silk fiber matrix. The wire rope design of the test model scaffold comprises several levels of geometric hierarchy. Commonly used degumming solutions fail in removing sericin in this wire rope design. Weight loss measurements, picric acid and carmine staining as well as scanning electron microscopy demonstrated that the removal of sericin from the model scaffold of a wire rope design can be achieved through a borate buffer-based system. Furthermore, the borate buffer degummed silks were shown to be nontoxic and did not alter cell proliferation behavior. The possibility to remove sericin after the textile engineering process has taken place eases the production of highly ordered scaffold structures and may expand the use of silk as scaffold material in further TE and regenerative medicine applications.

  7. Differential expression of early viral gene BmORF51 in Bombyx mori ...

    African Journals Online (AJOL)

    The transcription and protein product of early viral gene, Bm51, was detected at 6 h post-infection (p.i.) in resistant strain NB by quantitative real-time (qRT)-PCR and western blotting, and the expression of Bm51 in NB reached the maximal level at 36 h p.i. in NB, and then gradually decreased to undetectable level at 72 h ...

  8. The p10 gene of Bombyx mori nucleopolyhedrosis virus encodes a ...

    Indian Academy of Sciences (India)

    buffer, and subjected to electrophoresis on an 8% acrylamide. ± 6 M urea sequencing gel. For slot blot hybridization, 10"g total RNA isolated at different times following viral infection was blotted onto nylon membranes, UV-crosslinked and hybridized with the p10 antisense probe at 45 C for 12h. The membrane was washed ...

  9. Subcellular localization of Bombyx mori ribosomal protein S3a and ...

    African Journals Online (AJOL)

    USER

    2010-04-05

    Apr 5, 2010 ... piggybac-A3-EGFP and recombinant baculovirues expressing BmS3a-EGFP fusion protein (BV/IE1-. Bms3a-EGFP) or EGFP (BV/EGFP) were produced using BmNPV/Bac-to-Bac expression system. Results showed that the number of polyhedral in the transgenic cells of BmS3a was much smaller than.

  10. The Effect of Dietary Supplements on the Development of Bombyx Mori L. Silkworms

    Directory of Open Access Journals (Sweden)

    Cristina Zah

    2011-05-01

    Full Text Available We know that the silkworms consume leaves in large quantities. The scope of the research was their reaction to various additives for their food. Mulberry leaves spray-coated in several dietary supplements were administered starting with the 5th day of the 3rd instar. The substances used were flax (linseed oil, hemp oil and 2.5% fat cow’s milk. The research was performed on four different silkworm hybrid strands. Each hybrid was separated in 4 lots, a control group and one for each of the three supplements. The preliminary conclusions of the research were that the best results were obtained with the 2.5% milk supplement, where we observed the highest individual mass and silk quantity compared to the other lots.

  11. A novel Cph-like gene involved in histogenesis and maintenance of midgut in Bombyx mori.

    Science.gov (United States)

    Gan, Liping; Zhuo, Weiwei; Li, Juan; Wang, Ying; Sima, Yanghu; Xu, Shiqing

    2013-12-01

    Male-biased silkworm larva resistance is useful for sericulture and lepidopteran pest control. According to previous research, the mechanism underlying this resistance might be related to midgut-specific proteins. A midgut-specific and novel hypothetical cuticular protein-like (Cph-like) gene was cloned, based on sex-disparity serial analysis of gene expression (SAGE) libraries of the B. mori midgut. The complete cDNA contained 676 bp and encoded 165 amino acid residues. The gene was located on chromosome 19 and it had only one short 75 bp intron. The Cph-like expression level was downregulated by exogenous 20-hydroxyecdysone or starvation, but upregulated by exogenous methoprene or food intake. Knockdown (siRNA) of the Cph-like gene suppressed the appetite and delayed larval development, while it also degraded enterocytes and damaged the midgut morphology. Furthermore, the male-biased cytoplasmic polyhedrosis virus (BmCPV) resistance of larvae was decreased. The Cph-like gene is a midgut-specific novel gene in B. mori that may participate in histogenesis and midgut maintenance. © 2013 Society of Chemical Industry.

  12. Identification and Characterization of Bacillus cereus SW7-1 in Bombyx mori (Lepidoptera: Bombycidae).

    Science.gov (United States)

    Li, Guan-Nan; Xia, Xue-Juan; Zhao, Huan-Huan; Sendegeya, Parfait; Zhu, Yong

    2015-01-01

    The bacterial diseases of silkworms cause significant reductions in sericulture and result in huge economic loss. This study aimed to identify and characterize a pathogen from diseased silkworm. SW7-1, a pathogenic bacterial strain, was isolated from the diseased silkworm. The strain was identified on the basis of its bacteriological properties and 16S rRNA gene sequence. The colony was round, slightly convex, opaque, dry, and milky on a nutrient agar medium, the colony also exhibited jagged edges. SW7-1 was Gram-positive, without parasporal crystal, and 0.8-1.2 by 2.6-3.4 µm in length, resembling long rods with rounded ends. The strain was positive to most of the physiological biochemical tests used in this study. The strain could utilize glucose, sucrose, and maltose. The results of its 16S rRNA gene sequence analysis revealed that SW7-1 shared the highest sequence identity (>99%) with Bacillus cereus strain 14. The bacterial strain was highly susceptible to gentamycin, streptomycin, erythromycin, norfloxacin, and ofloxacin and moderately susceptible to tetracycline and rifampicin. It exhibited resistance to other antibiotics. SW7-1 had hemolytic activity and could produce extracellular casease, lipase, and amylase. SW7-1 could reproduce septicemia-like symptoms with high mortality rate when re-fed to healthy silkworm. .The median lethal concentration (LC50) was 5.45 × 10(4) cfu/ml. Thus, SW7-1 was identified as B. cereus, which is a pathogen for silkworm and human infections are possible. © The Author 2015. Published by Oxford University Press on behalf of the Entomological Society of America.

  13. Disruption of PTPS Gene Causing Pale Body Color and Lethal Phenotype in the Silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Xiaoling Tong

    2018-03-01

    Full Text Available Phenylketonuria (PKU is an inborn error of metabolism caused by mutations in the phenylalanine hydroxylase (PAH gene or by defects in the tetrahydrobiopterin (BH4 synthesis pathway. Here, by positional cloning, we report that the 6-pyruvoyl-tetrahydropterin synthase (PTPS gene, encoding a key enzyme of BH4 biosynthesis, is responsible for the alc (albino C mutation that displays pale body color, head shaking, and eventually lethality after the first molting in silkworm. Compared to wild type, the alc mutant produced more substrates (phenylalanine (Phe and tyrosine (Tyr and generated less DOPA and dopamine. Application of 2,4-diamino-6-hydroxypyrimidine (DAHP to block BH4 synthesis in the wild type effectively produced the alc-like phenotype, while BH4 supplementation rescued the defective body color and lethal phenotype in both alc and DAHP-treated individuals. The detection of gene expressions and metabolic substances after drugs treatments in alc and normal individuals imply that silkworms and humans have a high similarity in the drugs metabolic features and the gene pathway related to BH4 and the dopamine biosynthesis. We propose that the alc mutant could be used as an animal model for drug evaluation for BH4-deficient PKU.

  14. Interrelation between viability of silkworm larvae Bombyx mori and natural electromagnetic fields

    Directory of Open Access Journals (Sweden)

    V. M. Litvin

    2006-12-01

    Full Text Available The interrelation of viability of silkworm larvae with natural electromagnetic fields is investigated. On the large statistical material found by the UAAS Institute of Sericulture for 50 years, it is shown, that the geomagnetic activity is characterized by weak influence on viability of larvae populations (4,7 %. The greatest caterpillars’ viability is observed in years of a high level of geomagnetic activity.

  15. The adverse effects of phoxim exposure in the midgut of silkworm, Bombyx mori.

    Science.gov (United States)

    Gu, ZhiYa; Zhou, YiJun; Xie, Yi; Li, FanChi; Ma, Lie; Sun, ShanShan; Wu, Yu; Wang, BinBin; Wang, JuMei; Hong, Fashui; Shen, WeiDe; Li, Bing

    2014-02-01

    The silkworm is an important economic insect. Poisoning of silkworms by organophosphate pesticides causes tremendous loss to the sericulture. In this study, Solexa sequencing technology was performed to profile the gene expression changes in the midgut of silkworms in response to 24h of phoxim exposure and the impact on detoxification, apoptosis and immune defense were addressed. The results showed that 254 genes displayed at least 2.0-fold changes in expression levels, with 148 genes up-regulated and 106 genes down-regulated. Cytochrome P450 played an important role in detoxification. Histopathology examination and transmission electron microscope revealed swollen mitochondria and disappearance of the cristae of mitochondria, which are the important features in insect apoptotic cells. Cytochrome C release from mitochondria into the cytoplasm was confirmed. In addition, the Toll and immune deficiency (IMD) signal pathways were all inhibited using qRT-PCR. Our results could help better understand the impact of phoxim exposure on silkworm. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Structure and expression of the silk adhesive protein Ser2 in Bombyx mori

    Czech Academy of Sciences Publication Activity Database

    Kludkiewicz, Barbara; Takasu, Y.; Fedič, Robert; Tamura, T.; Sehnal, František; Žurovec, Michal

    2009-01-01

    Roč. 39, č. 12 (2009), s. 938-946 ISSN 0965-1748 R&D Projects: GA AV ČR IAA5007402; GA MŠk(CZ) LC06077 Institutional research plan: CEZ:AV0Z50070508 Keywords : silkworm * sericin * cocoon Subject RIV: CE - Biochemistry Impact factor: 3.117, year: 2009

  17. The p10 gene of Bombyx mori nucleopolyhedrosis virus encodes a ...

    Indian Academy of Sciences (India)

    E. L., Goldbach R. W. and Vlak J. M. 1993 Functional domains of the p10 protein of Autographa californica nuclear polyhe- drosis virus. J. Gen. Virol. 74, 563±574. Vlak J. M., Klinkenberg F. A., Zaal K. J. M., Usmany M., Klinge-. Roode E. C., Geervliet J. B. F., Roosien J. and Van Lent J. W. M.. 1988 Functional studies on the ...

  18. Insect-machine hybrid system for understanding and evaluating sensory-motor control by sex pheromone in Bombyx mori.

    Science.gov (United States)

    Kanzaki, Ryohei; Minegishi, Ryo; Namiki, Shigehiro; Ando, Noriyasu

    2013-11-01

    To elucidate the dynamic information processing in a brain underlying adaptive behavior, it is necessary to understand the behavior and corresponding neural activities. This requires animals which have clear relationships between behavior and corresponding neural activities. Insects are precisely such animals and one of the adaptive behaviors of insects is high-accuracy odor source orientation. The most direct way to know the relationships between neural activity and behavior is by recording neural activities in a brain from freely behaving insects. There is also a method to give stimuli mimicking the natural environment to tethered insects allowing insects to walk or fly at the same position. In addition to these methods an 'insect-machine hybrid system' is proposed, which is another experimental system meeting the conditions necessary for approaching the dynamic processing in the brain of insects for generating adaptive behavior. This insect-machine hybrid system is an experimental system which has a mobile robot as its body. The robot is controlled by the insect through its behavior or the neural activities recorded from the brain. As we can arbitrarily control the motor output of the robot, we can intervene at the relationship between the insect and the environmental conditions.

  19. Shotgun proteomic analysis of Bombyx mori brain: emphasis on regulation of behavior and development of the nervous system.

    Science.gov (United States)

    Wang, Guo-Bao; Zheng, Qin; Shen, Yun-Wang; Wu, Xiao-Feng

    2016-02-01

    The insect brain plays crucial roles in the regulation of growth and development and in all types of behavior. We used sodium dodecyl sulfate polyacrylamide gel electrophoresis and high-performance liquid chromatography - electron spray ionization tandem mass spectrometry (ESI-MS/MS) shotgun to identify the proteome of the silkworm brain, to investigate its protein composition and to understand their biological functions. A total of 2210 proteins with molecular weights in the range of 5.64-1539.82 kDa and isoelectric points in the range of 3.78-12.55 were identified. These proteins were annotated according to Gene Ontology Annotation into the categories of molecular function, biological process and cellular component. We characterized two categories of proteins: one includes behavior-related proteins involved in the regulation of behaviors, such as locomotion, reproduction and learning; the other consists of proteins related to the development or function of the nervous system. The identified proteins were classified into 283 different pathways according to KEGG analysis, including the PI3K-Akt signaling pathway which plays a crucial role in mediating survival signals in a wide range of neuronal cell types. This extensive protein profile provides a basis for further understanding of the physiological functions in the silkworm brain. © 2014 Institute of Zoology, Chinese Academy of Sciences.

  20. Transcriptome analysis of the epidermis of the purple quail-like (q-lp) mutant of silkworm, Bombyx mori

    Science.gov (United States)

    Xia, Dingguo; Tang, Shunming; Shen, Xingjia

    2017-01-01

    A new purple quail-like (q-lp) mutant found from the plain silkworm strain 932VR has pigment dots on the epidermis similar to the pigment mutant quail (q). In addition, q-lp mutant larvae are inactive, consume little and grow slowly, with a high death rate and other developmental abnormalities. Pigmentation of the silkworm epidermis consists of melanin, ommochrome and pteridine. Silkworm development is regulated by ecdysone and juvenile hormone. In this study, we performed RNA-Seq on the epidermis of the q-lp mutant in the 4th instar during molting, with 932VR serving as the control. The results showed 515 differentially expressed genes, of which 234 were upregulated and 281 downregulated in q-lp. BLASTGO analysis indicated that the downregulated genes mainly encode protein-binding proteins, membrane components, oxidation/reduction enzymes, and proteolytic enzymes, whereas the upregulated genes largely encode cuticle structural constituents, membrane components, transport related proteins, and protein-binding proteins. Quantitative reverse transcription PCR was used to verify the accuracy of the RNA-Seq data, focusing on key genes for biosynthesis of the three pigments and chitin as well as genes encoding cuticular proteins and several related nuclear receptors, which are thought to play key roles in the q-lp mutant. We drew three conclusions based on the results: 1) melanin, ommochrome and pteridine pigments are all increased in the q-lp mutant; 2) more cuticle proteins are expressed in q-lp than in 932VR, and the number of upregulated cuticular genes is significantly greater than downregulated genes; 3) the downstream pathway regulated by ecdysone is blocked in the q-lp mutant. Our research findings lay the foundation for further research on the developmental changes responsible for the q-lp mutant. PMID:28414820

  1. Induction of dominant lethal mutations by alkylating agnets in germ-cells of the silkworm, Bombyx mori

    International Nuclear Information System (INIS)

    Murota, Tetsuo; Murakami, Akio.

    1977-01-01

    The comparison of the intensity of activity was made by measuring radiation equivalent chemical (REC) dose in the experiment of the induction of dominant lethal mutation, using the germ cells of pupae five days before the moths will be hatched. The alkylating agents employed in the experiment are methyl methanesulfonate (MMS), ethyl methanesulfonate (EMS), diethyl sulfate (DSC) and mitomycine-C (MC). X-ray irradiation was employed in order to indicate the capability of inducing mutation of the alkylating agents with the radiation equivalent chemical dose (REC dose). The dose-hatchability curves for the alkylating agents showed sigmoidal fashion as observed in X-ray, regardless of germ cells. The REC value at LD (50) was estimated by comparing the relative mutagenic capability of these chemicals. In sperm, EMS and DES with concentration of 1.0 x 10 -7 M/g showed the same lethality as about 2.3 kR and 0.6 kR of X-ray. However, no significant reduction of embryonic lethality after the treatment of pupae with MC (up to 2.1 x 10 -7 M/g) and MMS (up to 1.0 x 10 -6 M/g) was observed. As the results, the order of mutagenic effectiveness was as follows: EMS>DES>MMS approximately equal to MC. When oocytes in the mid-pupae were treated with MMS, EMS and MC with concentration of 1.0 x 10 -7 M/g, MMS and EMS showed the same effects as 12.8 kR and 0.6 kR. Surprisingly, MC showed the same lethality as 232.3 kR. This extremely high sensitivity of oocytes to MC may be ascribed to the inhibiting effect of the drug on the meiotic division. (Iwakiri, K.)

  2. Isolation and characterization of sex chromosome rearrangements generating male muscle dystrophy and female abnormal oogenesis in the silkworm, Bombyx mori.

    Science.gov (United States)

    Fujii, T; Yokoyama, T; Ninagi, O; Kakehashi, K; Obara, Y; Nenoi, M; Ishikawa, T; Mita, K; Shimada, T; Abe, H

    2007-07-01

    In deletion-mapping of W-specific RAPD (W-RAPD) markers and putative female determinant gene (Fem), we used X-ray irradiation to break the translocation-carrying W chromosome (W( Ze )). We succeeded in obtaining a fragment of the W( Ze ) chromosome designated as Ze (W), having 3 of 12 W-RAPD markers (W-Bonsai, W-Yukemuri-S, W-Yukemuri-L). Inheritance of the Ze (W) fragment by males indicates that it does not include the Fem gene. On the basis of these results, we determined the relative positions of W-Yukemuri-S and W-Yukemuri-L, and we narrowed down the region where Fem gene is located. In addition to the Ze (W) fragment, the Z chromosome was also broken into a large fragment (Z(1)) having the +( sch ) (1-21.5) and a small fragment (Z(2)) having the +( od ) (1-49.6). Moreover, a new chromosomal fragment (Ze (W)Z(2)) was generated by a fusion event between the Ze (W) and the Z(2) fragments. We analyzed the genetic behavior of the Z(1) fragment and the Ze (W)Z(2) fragment during male (Z/Z(1) Ze (W)Z(2)) and female (Z(1) Ze (W)Z(2)/W) meiosis using phenotypic markers. It was observed that the Z(1) fragment and the Z or the W chromosomes separate without fail. On the other hand, non-disjunction between the Ze (W)Z(2) fragment and the Z chromosome and also between the Ze (W)Z(2) fragment and the W chromosome occurred. Furthermore, the females (2A: Z/Ze (W)Z(2)/W) and males (2A: Z/Z(1)) resulting from non-disjunction between the Ze (W)Z(2) fragment and the W chromosome had phenotypic defects: namely, females exhibited abnormal oogenesis and males were flapless due to abnormal indirect flight muscle structure. These results suggest that Z(2) region of the Z chromosome contains dose-sensitive gene(s), which are involved in oogenesis and indirect flight muscle development.

  3. Transcriptional stimulation via SC site of Bombyx sericin-1 gene through an interaction with a DNA binding protein SGF-3.

    Science.gov (United States)

    Matsuno, K; Takiya, S; Hui, C C; Suzuki, T; Fukuta, M; Ueno, K; Suzuki, Y

    1990-01-01

    Three protein binding sites have been identified in the upstream region of the sericin-1 gene. Two of them, SA and SC sites, have been known as putative cis-acting elements. Using synthetic oligonucleotides of these binding sites, it was found that silk gland factor-1 (SGF-1) binds to the SA site, and silk gland factor-3 (SGF-3) binds to the SC site but not to a mutated SC site, SCM. Tissue distribution of the two factors was different. SGF-3 is present abundantly in the middle silk gland (MSG) where the sericin-1 gene is transcribed specifically but is also present in other cell types, though in a much less concentration. SGF-1 is observed very abundantly in the two parts of silk gland, MSG and posterior silk gland (PSG), but not in other cells. Templates containing multimerized SA or SC sites at -39 of the sericin-1 gene promoter were tested in MSG nuclear extracts. The SC multimer strongly activated transcription, while the mutant SCM multimer did not. The SA multimer also gave a slight stimulation of transcription. These results suggest that SGF-3 stimulates transcription through an interaction with the SC site, and SGF-1 does so weakly through the SA site. Images PMID:2336359

  4. Bombyx mori silk protein films microprocessing with a nanosecond ultraviolet laser and a femtosecond laser workstation: theory and experiments

    Science.gov (United States)

    Lazare, S.; Sionkowska, A.; Zaborowicz, M.; Planecka, A.; Lopez, J.; Dijoux, M.; Louména, C.; Hernandez, M.-C.

    2012-01-01

    Laser microprocessing of several biopolymers from renewable resources is studied. Three proteinic materials were either extracted from the extracellular matrix like Silk Fibroin/Sericin and collagen, or coming from a commercial source like gelatin. All can find future applications in biomedical experimentation, in particular for cell scaffolding. Films of ˜hundred of microns thick were made by aqueous solution drying and laser irradiation. Attention is paid to the properties making them processable with two laser sources: the ultraviolet and nanosecond (ns) KrF (248 nm) excimer and the infrared and femtosecond (fs) Yb:KGW laser. The UV radiation is absorbed in a one-photon resonant process to yield ablation and the surface foaming characteristics of a laser-induced pressure wave. To the contrary, resonant absorption of the IR photons of the fs laser is not possible and does not take place. However, the high field of the intense I>˜1012 W/cm2 femtosecond laser pulse ionizes the film by the multiphoton absorption followed by the electron impact mechanism, yielding a dense plasma capable to further absorb the incident radiation of the end of the pulse. The theoretical model of this absorption is described in detail, and used to discuss the presented experimental effects (cutting, ablation and foaming) of the fs laser. The ultraviolet laser was used to perform simultaneous multiple spots experiments in which energetic foaming yields melt ejection and filament spinning. Airborne nanosize filaments "horizontally suspended by both ends" (0.25 μm diameter and 10 μm length) of silk biopolymer were observed upon irradiation with large fluences.

  5. Distribution of circadian clock-related proteins in the cephalic nervous system of the silkworm, Bombyx mori

    Czech Academy of Sciences Publication Activity Database

    Sehadová, Hana; Markova, E. P.; Sehnal, František; Takeda, M.

    2004-01-01

    Roč. 19, č. 6 (2004), s. 466-482 ISSN 0748-7304 Grant - others:JSPS(JP) 99L01205 Institutional research plan: CEZ:AV0Z5007907 Keywords : circadian rhythm * PER * CYC Subject RIV: ED - Physiology Impact factor: 2.979, year: 2004

  6. Purification and functional characterization of a protein: Bombyx mori human growth hormone like protein in silkworm pupa.

    Directory of Open Access Journals (Sweden)

    Jianqing Chen

    Full Text Available Human growth hormone (hGH is a peptide hormone secreted by eosinophils of the human anterior pituitary, and a regulatory factor for a variety of metabolic pathways. A 30-kD protein from the pupa stage of silkworm was detected by Western blotting and confirmed by immunoprecipitation based on its ability to bind to anti-hGH antibody. This protein, named BmhGH-like protein, was purified from fresh silkworm pupas through low-temperature homogenization, filtration, and centrifugation to remove large impurity particles. The supernatants were precipitated, resuspended, and passed through a molecular sieve. Further purification by affinity chromatography and two-dimensional electrophoresis resulted in pure protein for analysis by MS MALDI-TOF-MS analysis. An alignment with predicted proteins indicated that BmhGH-like protein consisted of two lipoproteins, which we named hGH-L1 and hGH-L2. These proteins belong to the β-trefoil superfamily, with β domains similar to the spatial structure of hGH. Assays with K562 cells demonstrated that these proteins could promote cell division in vitro. To further validate the growth-promoting effects, hGH-L2 was cloned from pupa cDNA to create recombinant silkworm baculovirus vBmNPV-hGH-L2, which was used to infect silkworm BmN cells at low titer. Flow cytometric analysis demonstrated that the protein shortened the G0/G1 phase of the cells, and enabled the cells to rapidly traverse the G1/S phase transition point to enter S phase and promote cell division. Discovery of hGH-like protein in silkworm will once again arouse people's interest in the potential medicinal value of silkworm and establish the basis for the development of new hormone drugs.

  7. Transcriptome analysis of the epidermis of the purple quail-like (q-lp) mutant of silkworm, Bombyx mori.

    Science.gov (United States)

    Wang, Pingyang; Qiu, Zhiyong; Xia, Dingguo; Tang, Shunming; Shen, Xingjia; Zhao, Qiaoling

    2017-01-01

    A new purple quail-like (q-lp) mutant found from the plain silkworm strain 932VR has pigment dots on the epidermis similar to the pigment mutant quail (q). In addition, q-lp mutant larvae are inactive, consume little and grow slowly, with a high death rate and other developmental abnormalities. Pigmentation of the silkworm epidermis consists of melanin, ommochrome and pteridine. Silkworm development is regulated by ecdysone and juvenile hormone. In this study, we performed RNA-Seq on the epidermis of the q-lp mutant in the 4th instar during molting, with 932VR serving as the control. The results showed 515 differentially expressed genes, of which 234 were upregulated and 281 downregulated in q-lp. BLASTGO analysis indicated that the downregulated genes mainly encode protein-binding proteins, membrane components, oxidation/reduction enzymes, and proteolytic enzymes, whereas the upregulated genes largely encode cuticle structural constituents, membrane components, transport related proteins, and protein-binding proteins. Quantitative reverse transcription PCR was used to verify the accuracy of the RNA-Seq data, focusing on key genes for biosynthesis of the three pigments and chitin as well as genes encoding cuticular proteins and several related nuclear receptors, which are thought to play key roles in the q-lp mutant. We drew three conclusions based on the results: 1) melanin, ommochrome and pteridine pigments are all increased in the q-lp mutant; 2) more cuticle proteins are expressed in q-lp than in 932VR, and the number of upregulated cuticular genes is significantly greater than downregulated genes; 3) the downstream pathway regulated by ecdysone is blocked in the q-lp mutant. Our research findings lay the foundation for further research on the developmental changes responsible for the q-lp mutant.

  8. Lead in the soil-mulberry (Morus alba L.)-silkworm (Bombyx mori) food chain: translocation and detoxification.

    Science.gov (United States)

    Zhou, Lingyun; Zhao, Ye; Wang, Shuifeng; Han, Shasha; Liu, Jing

    2015-06-01

    The translocation of lead (Pb) in the soil-mulberry-silkworm food chain and the process of Pb detoxification in the mulberry-silkworm chain were investigated. The amount of Pb in mulberry, silkworm, feces and silk increased in a dose-responsive manner to the Pb contents in the soils. Mulberry roots sequestered most of the Pb, ranging from 230.78 to 1209.25 mg kg(-1). Over 92% of the Pb in the mulberry leaves was deposited in the cell wall, and 95.29-95.57% of the Pb in the mulberry leaves was integrated with oxalic acid, pectates and protein, and it had low bioavailability. The Pb concentrations in the silkworm feces were 4.50-4.64 times higher than those in the leaves. The synthesis of metallothioneins in three tissues of the silkworms was induced to achieve Pb homeostasis under Pb stress. These results indicated the mechanism involved in Pb transfer along the food chain was controlled by the detoxification of Pb in different trophic levels. Planting mulberry and rearing silkworm could be a promising approach for the remediation of Pb-polluted soils due to the Pb tolerance of mulberry and silkworm. Copyright © 2015. Published by Elsevier Ltd.

  9. Extensive conserved synteny of genes between the karyotypes of Manduca sexta and Bombyx mori revealed by BAC-FISH mapping

    Czech Academy of Sciences Publication Activity Database

    Yasukochi, Y.; Tanaka-Okuyama, M.; Shibata, F.; Yoshido, A.; Marec, František; Wu, Ch.; Zhang, H.; Goldsmith, M. R.; Sahara, K.

    2009-01-01

    Roč. 4, č. 10 (2009), e7465 E-ISSN 1932-6203 R&D Projects: GA ČR GA206/06/1860; GA AV ČR IAA600960925 Grant - others: Japan Society for the Promotion of Science(JP) 18380037; National Science Foundation(US) IBN020838 Institutional research plan: CEZ:AV0Z50070508 Keywords : Manduca sexta Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.351, year: 2009

  10. In vitro conditions for 14C-leucine incorporation into the protein of cultured ovaries of the silkworm, Bombyx mori

    International Nuclear Information System (INIS)

    Miyadai, Toshiaki; Yamashita, Okitsugu

    1980-01-01

    Vitellogenic ovaries of silkworm pupae were incubated in vitro in different media based on the Wyatt's medium to establish an adequate condition for culture of silkworm ovaries. Incorporation of 14 C-leucine into protein fraction was determined to assess the biochemical activity of the ovary. When ovaries were incubated in vitro for a short time by 6 hr, a saturation kinetics of incorporation of the labelled leucine was shown. Sequential substitution of K + ion to Na + ion in the medium had no effect on the incorporation of 14 C-leucine, but Mg 2+ ion appeared to stimulate synthetic activity at more than 10 mM. The activity was not affected at pH range 5.0-7.2. Neither different sugars, nor vitellogenin nor lipoprotein prepared from silkworm haemolymph affected the incorporation of 14 C-leucine, when added into the medium. The synthesis of protein depended upon the developmental stages of the cultured ovaries and was most active in 6-day-old ovary. Ovaries developing in pupal body showed comparable changes in synthetic activity. It is concluded that the chemical composition of the medium does not exert a strict effect on synthetic activity of protein in short-term cultures and the ovaries cultured in vitro maintain the activity comparable with those found in in situ condition. (author)

  11. Effect of solvents on properties of Bombyx mori silk grafted by methyl methacrylate (MMA and methacrylamide (MAA

    Directory of Open Access Journals (Sweden)

    Wattana Klairatsamee

    2005-11-01

    Full Text Available Mulberry silks were chemically modified in order to increase weight gain, resulting from degumming process using graft copolymerisation technique with vinyl monomers, i.e. MMA, MAA and MMA/MAA. Due to the appearance of PMMA homopolymer granules adhered on the MMA- and MMA/MAA-grafted silk surfaces resulting in surface roughness when silk was grafted by MMA in water, the influence of grafting solvents was examined, using different water/ethanol volume ratios of 100/0, 75/25, 50/50, 25/75 and 0/100. FTIR spectra of the grafted silks presented the absorption bands of the vinyl monomers used for the grafting process. In addition, high values of % polymer add-on were obtained for all of the grafted silks. It was also found that the suitable solvents were 25/75 water/ethanol for the silk grafted by MMA and MMA/MAA, and water for the silk grafted by MAA, in order to get the smooth grafted silk surface and high polymer add-on. Moreover, all the grafted silks showed slightly greater stiffness, as indicated by the increase of Young's modulus and the decrease of elongation.

  12. Determination of Albendazole and Metabolites in Silkworm Bombyx mori Hemolymph by Ultrafast Liquid Chromatography Tandem Triple Quadrupole Mass Spectrometry

    Science.gov (United States)

    Li, Li; Xing, Dong-Xu; Li, Qing-Rong; Xiao, Yang; Ye, Ming-Qiang; Yang, Qiong

    2014-01-01

    Albendazole is a broad-spectrum parasiticide with high effectiveness and low host toxicity. No method is currently available for measuring albendazole and its metabolites in silkworm hemolymph. This study describes a rapid, selective, sensitive, synchronous and reliable detection method for albendazole and its metabolites in silkworm hemolymph using ultrafast liquid chromatography tandem triple quadrupole mass spectrometry (UFLC-MS/MS). The method is liquid-liquid extraction followed by UFLC separation and quantification in an MS/MS system with positive electrospray ionization in multiple reaction monitoring mode. Precursor-to-product ion transitions were monitored at 266.100 to 234.100 for albendazole (ABZ), 282.200 to 208.100 for albendazole sulfoxide (ABZSO), 298.200 to 159.100 for albendazole sulfone (ABZSO2) and 240.200 to 133.100 for albendazole amino sulfone (ABZSO2-NH2). Calibration curves had good linearities with R2 of 0.9905–0.9972. Limits of quantitation (LOQs) were 1.32 ng/mL for ABZ, 16.67 ng/mL for ABZSO, 0.76 ng/mL for ABZSO2 and 5.94 ng/mL for ABZSO2-NH2. Recoveries were 93.12%–103.83% for ABZ, 66.51%–108.51% for ABZSO, 96.85%–105.6% for ABZSO2 and 96.46%–106.14% for ABZSO2-NH2, (RSDs <8%). Accuracy, precision and stability tests showed acceptable variation in quality control (QC) samples. This analytical method successfully determined albendazole and its metabolites in silkworm hemolymph in a pharmacokinetic study. The results of single-dose treatment suggested that the concentrations of ABZ, ABZSO and ABZSO2 increased and then fell, while ABZSO2-NH2 level was low without obvious change. Different trends were observed for multi-dose treatment, with concentrations of ABZSO and ABZSO2 rising over time. PMID:25255321

  13. Effect of gamma-rays on some technologic properties of cocoon and filament of the silk worm (Bombyx mori L.)

    International Nuclear Information System (INIS)

    Binkh, T.T.; Petkov, N.; Malinova, K.

    1993-01-01

    The effect of irradiation of silk worm eggs with 1.00, 2.00, 3.00, 4.00, 5.00, 6.00 and 7.00 Gy gamma-rays on some major technologic properties of cocoons and filament of the mulbery silk worm was studied. Single irradiation of silk worm during the phase of highest lengthening of nucleus development causes both stimulating and inhibitory effect on the technologic properties of the cocoons and the filament. The degree of stimulation or inhibition is dependent on the irradiation dose, the specificity of technologic processes and biologic characteristics of silk worm species and hybrids. Gamma-irradiation with doses 2.00 and 3.00 Gy stimulates the increase in weight of silk covering by 2.07-8.28%, raw cocoon silkness by 0.38-3.21 points and length of cocoon filament by 40-151 m. Irradiation of silk worm eggs with 2.00-4.00 Gy gamma-rays decreases by 1.4-2.3 kg (12.61-21.91%) the quantity of raw cocoons, required for production of 1 kg grege. 9 refs., 4 tabs. (author)

  14. γ-ray-induced mutations of female primordial germ-cells during organogenesis in the silkworm (Bombyx mori L.)

    International Nuclear Information System (INIS)

    Miki, Mutsuo; Murakami, Akio.

    1979-01-01

    Developing silkworm embryos from 44 up 212 hr after oviposition were irradiated with a constant single dose of 1000 R of 137 Cs gamma rays (dose-rate at 300 R per min) and the change of the female primordial germ-cells in the sensitivity to mutation induction during the organogenesis. Egg-color specific locus method was used for the detection of the mutational incidences. The results obtained were summarized as follows: The mutation frequency increased gradually with the development of gonad in embryos from 68 to 116 hr after oviposition and there decreased until 188 hr when the mutation frequency was as low as in the 44 hr old embryo. The frequency increased slightly at 212 hr. The number of germ-cells increased at a rate of 30 hr/generation during the gonad formation in embryos from 68 to 128 hr after oviposition. Gonads were complete in 116 hr-old embryos. The multiplication of primordial germ-cells ceased temporarily after blastokinesis until the onset of gonial divisions in aged embryos 2 days before hatching. The change of primordial germ-cells in radiosensitivity appeared to depend on whether the germ-cells were in the process of multiplication. (author)

  15. Evaluation of genetic potential of the polyvoltine silkworm (Bombyx mori L.) germplasm and identification of parents for breeding programme*

    Science.gov (United States)

    Rao, C.G.P.; Seshagiri, S.V.; Ramesh, C.; Ibrahim, Basha K.; Nagaraju, H.; Chandrashekaraiah

    2006-01-01

    In the present study, polyvoltine germplasm stock of Andhra Pradesh State Sericulture Research and Development, Institute (APSSRDI) was evaluated for its performance based on quantitative and qualitative traits. Twenty-one oval and 10 peanut cocoon shaped lines were reared in different seasons of the year. Since the polyvoltines are non-diapausing, six generations were reared and evaluated for various economically important traits based on evaluation index and sub-ordinate function statistical methods. Ten top ranked lines obtained by using both the methods were identified as potential parental strains. Among oval lines, APM14, APM11, APM18, APMW9, and APM19, and among peanut lines APMD5, APMD1, APMD3, APMD9 and APMD8 were selected as base material. The identified high yielding lines will be used in various breeding programmes as initial parents for the synthesis of superior polyvoltine breeds/hybrids. PMID:16502509

  16. In vivo analysis of fibroin heavy chain signal peptide of silkworm Bombyx mori using recombinant baculovirus as vector

    International Nuclear Information System (INIS)

    Wang Shengpeng; Guo Tingqing; Guo Xiuyang; Huang Junting; Lu Changde

    2006-01-01

    In order to investigate the functional signal peptide of silkworm fibroin heavy chain (FibH) and the effect of N- and C-terminal parts of FibH on the secretion of FibH in vivo, N- and C-terminal segments of fibh gene were fused with enhanced green fluorescent protein (EGFP) gene. The fused gene was then introduced into silkworm larvae and expressed in silk gland using recombinant AcMNPV (Autographa californica multiple nuclear polyhedrosis virus) as vector. The fluorescence of EGFP was observed with fluorescence microscope. FibH-EGFP fusion proteins extracted from silk gland were analyzed by Western blot. Results showed that the two alpha helices within N-terminal 163 amino acid residues and the C-terminal 61 amino acid residues were not necessary for cleavage of signal peptide and secretion of the fusion protein into silk gland. Then the C-terminal 61 amino acid residues were substituted with a His-tag in the fusion protein to facilitate the purification. N-terminal sequencing of the purified protein showed that the signal cleavage site is between position 21 and 22 amino acid residues

  17. The effect of gamma irradiation on some morphological and quantitative changes of Bombyx mori L. silk gland

    International Nuclear Information System (INIS)

    Petkov, N.; Malinova, K.; Binkh, N.

    1996-01-01

    A study to determine the effect of gamma irradiation of silkworm eggs at doses of 1.00; 2.00 and 3.00 on silk gland weight, length and width is performed. It was found that gamma irradiation of eggs in the stage of embryo most intensive growth in length (B 2 ) at doses 2.00 and 3.00 Gy stimulates increasing of silk glands weight in silkworms on the fifth day instar by 12-25 mg, as well as the silk worm width - by 7-33 μm, which is of significant importance for the synthesis and secretion of silk proteins (fibroin and sericin). A breed specificity was also observed

  18. A study on the spreading ability of radiation-sterilized male moth of bombyx mandarin in field

    International Nuclear Information System (INIS)

    Yang Rongxin; Xia Darong; Gu Weiping; Zhang Yanjun

    1998-01-01

    The mulberry wild silkworm (MWS), belong to the Bombycidae of Lepidoptera, is serious pest of sericulture. The female moth of MWS is sterile and the male moth is sub-sterile when they were treated with 250 Gy 60 Co γ-ray (dose rate: 1.05 Gy/min), and their filial generations was sterile. The spreading ability of male moth of MWS in field and retrieving the marked MWS male moth with the trapping method was studied. The trapping solution was composed of sugar, vinegar, wine and alive female moth. The retrieving rate of MWS male moth amounted to 12.6%∼13.5% of released moth in field. The spreading range in 24 hours for sterile MWS male moth reached to 700 m, and 90.8% of MWS male moth was in an area of 5 m radius from the releasing centre. It is concluded that thirty releasing centres per hectare are needed to make the irradiated sterilized insects spread for controlling the MWS in field

  19. Isolation of a mutant line with shorter larval duration by induction of mutations in the silkworm Bombyx mori L

    International Nuclear Information System (INIS)

    Subramanya, G.; Sreerama Reddy, G.

    1982-01-01

    Induction of mutations by X-rays in the pure Mysore race of the silkworm, B. mori, has resulted in the isolation of a mutant line exhibiting a much shorter larval duration than the control. The 8th day eggs and 3rd instar larvae were exposed to variable doses of X-rays of 1000 and 2000 R respectively. The 8th day eggs were found to be more sensitive for irradiation. The mutant line isolated from the progenies of the irradiated eggs exhibits a relatively faster rate of development. In addition, the mutant line exhibits certain degree of improvement in the commercial characters such as cocoon weight, shell weight percentage of silk, etc. over those of controls. The significance of faster rate of development and improvement in the commercial characters of the mutant line are discussed in relation to economics of sericulture industry. (author)

  20. Study on the effect of gamma-radiation on the development of silk worm (Bombyx mori L.)

    International Nuclear Information System (INIS)

    Grekov, D.

    1995-01-01

    These investigations are of special interest to the breeding practice considering the lack of profound research activity. The main purpose was to find out possibilities of using gamma-rays as a mutagenic factor in sericulture. The results obtained show that the irradiation of silk worm eggs at b 2 stage of embryo development with 5 Gy stimulates the biological development. The irradiation doses of 10-25 Gy inhibit mulberry silk worm development. The different species react in different way. Probably mutation forms are to be sought within 10-25 Gy doses. 10 refs., 3 tabs. (author)

  1. Isolation of a mutant line with shorter larval duration by induction of mutations in the silkworm Bombyx mori L

    Energy Technology Data Exchange (ETDEWEB)

    Subramanya, G.; Sreerama Reddy, G.

    1982-02-01

    Induction of mutations by X-rays in the pure Mysore race of the silkworm, B. mori, has resulted in the isolation of a mutant line exhibiting a much shorter larval duration than the control. The 8th day eggs and 3rd instar larvae were exposed to variable doses of X-rays of 1000 and 2000 R respectively. The 8th day eggs were found to be more sensitive for irradiation. The mutant line isolated from the progenies of the irradiated eggs exhibits a relatively faster rate of development. In addition, the mutant line exhibits a certain degree of improvement in the commercial characters such as cocoon weight, shell weight percentage of silk, etc. over those of controls. The significance of faster rate of development and improvement in the commercial characters of the mutant line are discussed in relation to economics of sericulture industry.

  2. Mechanisms of TiO2 NPs-induced phoxim metabolism in silkworm (Bombyx mori) fat body.

    Science.gov (United States)

    Hu, J S; Li, F C; Xu, K Z; Ni, M; Wang, B B; Tian, J H; Li, Y Y; Shen, W D; Li, B

    2016-05-01

    Silkworm is an important economic insect. Abuse of organophosphorus pesticides in recent years often leads to poisoning of silkworms, which significantly affects sericulture development by reducing silk production. Previous studies have shown that TiO2 NPs can effectively mitigate the damages caused by organophosphorus pesticides in silk glands and nerve tissues. The fat body is an important metabolic detoxification organ of silkworms, but it is unknown whether TiO2 NPs affect pesticide metabolism in fat body. In this study, we characterized the transcription of antioxidant genes and enzyme activity in fat body after TiO2 NPs and phoxim treatments using transcriptome sequencing, real-time PCR, and enzyme activity assay. Transcriptome sequencing detected 10 720, 10 641, 10 403, and 10 489 genes for control group, TiO2 NPs group, phoxim group, and TiO2 NPs+phoxim group, respectively. The TiO2 NPs+phoxim group had 705 genes with significantly differential expression (FDR<0.001), among which the antioxidant genes thioredoxin reductase 1 and glutathione S-transferase omega 3 were significantly upregulated. In phoxim group, the expression levels of superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase delta (GSTd), and thioredoxin peroxidase (TPx) were increased by 1.365 -fold, 1.335 -fold, 1.642 -fold, and 1.765 -fold, respectively. The level changes of SOD, CAT, GSTd, and TPx were validated by real time PCR. The contents of reactive oxygen species (ROS), malondialdehyde (MDA), and hydrogen peroxide (H2O2) were increased by 1.598 -fold, 1.946 -fold, and 1.506 -fold, respectively, indicating that TiO2 NPs treatment can relieve phoxim-induced oxidative stress. To clarify the mechanism of TiO2 NPs's effect, the transcription levels of P450 gene family were measured for the TiO2 NPs+phoxim group; the expression levels of CYP4M5, CYP6AB4, CYP6A8, and CYP9G3 were elevated by 2.784 -fold, 3.047 -fold, 2.254 -fold, and 4.253 -fold, respectively, suggesting that high expression of P450 family genes can enhance the metabolism of phoxim in the fat body. The results of this study indicated that TiO2 NPs treatment promoted the transcriptional expression of the P450 family genes to improve the fat body's ability to metabolize phoxim and reduce phoxim-induced oxidative stress. This may be the main mechanism of TiO2 NPs' mitigation of phoxim-induced damages in the fat body. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Preparation and Characterization of a Novel Hybrid Hydrogel Composed of Bombyx mori Fibroin and Poly(N-isopropylacrylamide

    Directory of Open Access Journals (Sweden)

    Ting Wang

    2013-01-01

    Full Text Available A novel hybrid hydrogel was prepared and investigated based on silkworm silk fibroin and poly(N-isopropylacrylamide (PNIPAAm. PNIPAAm was introduced to silk fibroin, the resultant composite hydrogel was examined, and freeze-dried SF/PNIPAAm scaffold was analyzed using LB-550 dynamic light scattering particle-size analyzer, circular dichroism (CD, and scanning electron microscopy (SEM. Our results suggested that the hybrid hydrogels owned the porous sponge-like structures, and the gelation time of SF/PNIPAAm hybrids decreased with an increase in temperature and concentration of each polymer. Results of rheological analysis suggested that the rheological property of resultant SF/PNIPAAm gel depended on the concentration combinations as well as the aging time, which elapsed after mixing the two polymers. Results of CD spectra demonstrated that pH showed little influence on the secondary structure of silk fibroin, and significant changes of , , and G* as surrounding increase temperature above the lower critical solution temperature (LCST.

  4. 1849-IJBCS-Article-Sunday Adeduntan

    African Journals Online (AJOL)

    hp

    eggs and hatchability of eggs) of silkworm. MATERIALS AND METHODS. Procurement and biovoltine strain Bombyx mori. The biovoltine strain of Bombyx mori was obtained from the sericulture centre of. Ondo State 9 dfls (diseases free lay) of ND18 race of silkworm (Bombyx mori) and three varieties of mulberry plant used ...

  5. On the Breeding of Bivoltine Breeds of the Silkworm, Bombyx mori L. (Lepidoptera: Bombycidae, Tolerant to High Temperature and High Humidity Conditions of the Tropics

    Directory of Open Access Journals (Sweden)

    Harjeet Singh

    2010-01-01

    Full Text Available The hot climatic conditions of tropics prevailing particularly in summer are contributing to the poor performance of the bivoltine breeds and the most important aspect is that many quantitative characters such as viability and cocoon traits decline sharply when temperature is high. Hence, in a tropical country like India, it is very essential to develop bivoltine breeds/hybrids which can withstand the high temperature stress conditions. This has resulted in the development of CSR18 × CSR19, compatible hybrid for rearing throughout the year by utilizing Japanese thermotolerant hybrids as breeding resource material. Though, the introduction of CSR18 × CSR19 in the field during summer months had considerable impact, the productivity level and returns realized do not match that of other productive CSR hybrids. Therefore, the acceptance level of this hybrid with the farmers was not up to the expected level. This has necessitated the development of a temperature tolerant hybrid with better productivity traits than CSR18 × CSR19. Though, it was a difficult task to break the negative correlation associated with survival and productivity traits, attempts on this line had resulted in the development of CSR46 × CSR47, a temperature tolerant bivoltine hybrid with better productivity traits than CSR18 × CSR19. However, though, these hybrids are tolerant to high temperature environments, they are not tolerant to many of the silkworm diseases. Keeping this in view, an attempt is made to develop silkworm hybrids tolerant to high temperature environments.

  6. Precise genome editing in the silkworm Bombyx mori using TALENs and ds- and ssDNA donors – A practical approach

    Czech Academy of Sciences Publication Activity Database

    Takasu, Y.; Kobayashi, I.; Tamura, T.; Uchino, K.; Sezutsu, H.; Žurovec, Michal

    2016-01-01

    Roč. 78, NOV 01 (2016), s. 29-38 ISSN 0965-1748 R&D Projects: GA ČR GA14-27816S Institutional support: RVO:60077344 Keywords : engineered nucleases * HDR * DSB Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.756, year: 2016 http://www.sciencedirect.com/science/article/pii/S0965174816301242

  7. A proteomic view on the developmental transfer of homologous 30 kDa lipoproteins from peripheral fat body to perivisceral fat body via hemolymph in silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Pakkianathan Britto

    2012-02-01

    Full Text Available Abstract Background A group of abundant proteins of ~30 kDa is synthesized in silkworm larval peripheral fat body (PPFB tissues and transported into the open circulatory system (hemolymph in a time-depended fashion to be eventually stored as granules in the pupal perivisceral fat body (PVFB tissues for adult development during the non-feeding stage. These proteins have been shown to act anti-apoptotic besides being assigned roles in embryogenesis and defense. However, detailed protein structural information for individual PPFB and PVFB tissues during larval and pupal developmental stages is still missing. Gel electrophoresis and chromatography were used to separate the 30 kDa proteins from both PPFB and PVFB as well as hemolymph total proteomes. Mass spectrometry (MS was employed to elucidate individual protein sequences. Furthermore, 30 kDa proteins were purified and biochemically characterized. Results One- and two-dimensional gel electrophoresis (1/2D-PAGE was used to visualize the relative changes of abundance of the 30 kDa proteins in PPFB and PVFB as well as hemolymph from day 1 of V instar larval stage to day 6 of pupal stage. Their concentrations were markedly increased in hemolymph and PVFB up to the first two days of pupal development and these proteins were consumed during development of the adult insect. Typically, three protein bands were observed (~29, 30, 31 kDa in 1D-PAGE, which were subjected to MS-based protein identification along with spots excised from 2D-gels run for those proteomes. Gas phase fragmentation was used to generate peptide sequence information, which was matched to the available nucleotide data pool of more than ten highly homologous insect 30 kDa lipoproteins. Phylogenetic and similarity analyses of those sequences were performed to assist in the assignment of experimentally identified peptides to known sequences. Lipoproteins LP1 to LP5 and L301/302 could be matched to peptides extracted from all bands suggesting the presence of full length and truncated or modified protein forms in all of them. The individual variants could not be easily separated by classical means of purification such as 2D-PAGE because of their high similarity. They even seemed to aggregate as was indicated by native gel electrophoresis. Multistep chromatographic procedures eventually allowed purification of an LP3-like protein. The protein responded to lipoprotein-specific staining. Conclusions In B. mori larvae and pupae, 30 kDa lipoproteins LP1 to LP5 and L301/302 were detected in PPFB and PVFB tissue as well as in hemolymph. The concentration of these proteins changed progressively during development from their synthesis in PPFB, transport in hemolymph to storage in PVFB. While the 30 kDa proteins could be reproducibly separated in three bands electrophoretically, the exact nature of the individual protein forms present in those bands remained partially ambiguous. The amino acid sequences of all known 30 kDa proteins showed very high homology. High-resolution separation techniques will be necessary before MS and other structural analysis can shed more light on the complexity of the 30 kDa subproteome in B. mori. A first attempt to that end allowed isolation of a B. mori LP3-like protein, the complete structure, properties and function of which will now be elucidated in detail.

  8. Identification of MBF2 family genes in Bombyx mori and their expression in different tissues and stages and in response to Bacillus bombysepticus infection and starvation.

    Science.gov (United States)

    Zhou, Chun-Yan; Zha, Xing-Fu; Liu, Chun; Han, Min-Jin; Zhang, Li-Ying; Shi, Pan-Pan; Wang, He; Zheng, Ren-Wen; Xia, Qing-You

    2016-08-01

    The Multiprotein bridge factor 2 (MBF2) gene was first identified as a co-activator involved in BmFTZ-F1-mediated activation of the Fushi tarazu gene. Herein, nine homologous genes of MBF2 gene are identified. Evolutionary analysis showed that this gene family is insect-specific and that the family members are closely related to response to pathogens (REPAT) genes. Tissue distribution analysis revealed that these genes could be expressed in a tissue-specific manner. Developmental profiles analysis showed that the MBF2 gene family members were highly expressed in the different stages. Analysis of the expression patterns of nine MBF2 family genes showed that Bacillus bombysepticus treatment induced the up-regulation of several MBF2 family genes, including MBF2-4, -7, -9, -8. Furthermore, we found the MBF2 family genes were modulated by starvation and the expression of these genes recovered upon re-feeding, except for MBF2-5, -9. These findings suggested roles for these proteins in insect defense against pathogens and nutrient metabolism, which has an important guiding significance for designing pest control strategies. © 2016 Institute of Zoology, Chinese Academy of Sciences.

  9. Antioxidant activities of two sericin proteins extracted from cocoon of silkworm (Bombyx mori) measured by DPPH, chemiluminescence, ORAC and ESR methods

    OpenAIRE

    TAKECHI, TAYORI; WADA, RITSUKO; FUKUDA, TSUBASA; HARADA, KAZUKI; TAKAMURA, HITOSHI

    2014-01-01

    Recent efforts have focused on the use of sericin proteins extracted from cocoons of silkworm as a healthy food source for human consumption. In this study, we focused on the antioxidative properties of sericin proteins. The antioxidative properties were measured in sericin proteins extracted from the shell of the cocoon, designated hereafter as white sericin protein and yellow-green sericin protein, as well as bread without sericin protein and bread to which white sericin powder had been add...

  10. On the Effects of Juvenile Hormone and Ecdysterone for the Parthenogenetic Occurrence in the Silkworm, Bombyx mori L.(Lepi-d optera: Bombycidae)

    OpenAIRE

    TAKEI, Ryuzo; ONO, Mikiko

    2002-01-01

    The morphological change of an egg cell and the manifestation state of a parthenogenetic egg were investigated by administering Juvenile hormone and ecdysterone being endocrine hormone at a pupal stage. The results obtained are as follows. 1) The manifestation rate of a parthenogenetic eggs was highest in an ecdysterone administration group and lowered in a control group and became lowest in a juvenile hormone administration group. 2) In view of the morphological change of an egg cell, the mi...

  11. BmCHSA-2b, a Lepidoptera specific alternative splicing variant of epidermal chitin synthase, is required for pupal wing development in Bombyx mori.

    Science.gov (United States)

    Xu, Guanfeng; Zhang, Jie; Lyu, Hao; Liu, Jia; Ding, Yang; Feng, Qili; Song, Qisheng; Zheng, Sichun

    2017-08-01

    Insect chitin synthase A (CHSA) is an epidermis-specific enzyme that plays an essential role in insect development. In this study, the function and regulation of CHSA-2b, an alternative splicing variant of Bombxy mori CHSA that is discovered only in Lepidopteran insects, were investigated. Analysis of mRNA level showed that BmCHSA-2b was responsive to 20-hydroxyecdysone (20E) in pupal wing unlike BmCHSA-2a, which shares almost the identical sequence as BmCHSA-2b except the first 31 amino acids, suggesting that the expression of these two alternative splicing variants is driven by different promoters of CHSA gene. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) analysis showed that BmCHSA-2b was up-regulated in the wing of mid-pupa unlike BmCHSA-2a, which was up-regulated in epidermis and wing disc at the beginning and end of pupal stage. Further analysis reveals that the up-regulations of BmCHSA-2a and BmCHSA-2b in pupal wing were consistent with the increase of chitin content and wing area at the same stages, respectively. Furthermore, the higher transcription level of BmCHSA-2b in the mid-pupal wing of male than that in female was consistent with the chitin content of pupal wing between genders. Injection of double-stranded RNAs of BmCHSA-2b resulted in the decrease in the area and chitin content of the wing, and irregular and crimpled vein. All these results together suggest that B. mori evolves an extra promoter in CHSA gene to activate BmCHSA-2b expression in the wing of mid-pupal stage in response to 20E, and BmCHSA-2b is required for the wing development in the mid-pupa of B. mori. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. DNA Synthesis during Endomitosis Is Stimulated by Insulin via the PI3K/Akt and TOR Signaling Pathways in the Silk Gland Cells of Bombyx mori

    Directory of Open Access Journals (Sweden)

    Yaofeng Li

    2015-03-01

    Full Text Available Silk gland cells undergo multiple endomitotic cell cycles during silkworm larval ontogeny. Our previous study demonstrated that feeding is required for continued endomitosis in the silk gland cells of silkworm larvae. Furthermore, the insulin signaling pathway is closely related to nutritional signals. To investigate whether the insulin signaling pathway is involved in endomitosis in silk gland cells, in this study, we initially analyzed the effects of bovine insulin on DNA synthesis in endomitotic silk gland cells using 5-bromo-2'-deoxyuridine (BrdU labeling technology, and found that bovine insulin can stimulate DNA synthesis. Insulin signal transduction is mainly mediated via phosphoinositide 3-kinase (PI3K/Akt, the target of rapamycin (TOR and the extracellular signal-regulated kinase (ERK pathways in vertebrates. We ascertained that these three pathways are involved in DNA synthesis in endomitotic silk gland cells using specific inhibitors against each pathway. Moreover, we investigated whether these three pathways are involved in insulin-stimulated DNA synthesis in endomitotic silk gland cells, and found that the PI3K/Akt and TOR pathways, but not the ERK pathway, are involved in this process. These results provide an important theoretical foundation for the further investigations of the mechanism underlying efficient endomitosis in silk gland cells.

  13. Changes in diapause related gene expression pattern during early embryonic development in HCl-treated eggs of bivoltine silkworm Bombyx mori (Lepidoptera: Bombycidae

    Directory of Open Access Journals (Sweden)

    Sirigineedi Sasibhushan

    2013-02-01

    Full Text Available Investigation of differential expression of diapause related genes (five metabolic, five heat shock protein and one translational regulatory in HCl-treated (non-diapause and untreated (diapause eggs of B. mori during early embryogenesis (up to 48h following oviposition revealed the up-regulation of sorbitol dehydrogenase upon HCl treatment, indicating increased glycogen synthesis for further embryonic development but, down-regulation of phosphofructo kinase gene expression after 18h of oviposition indicating an arrest of glycerol and sorbitol conversion. The expression of poly A binding protein gene expression was higher upon HCl treatment, revealing the initiation of translation. The expression levels of other genes analyzed did not vary significantly, except for Hsp90 and Hsp40, which were up-regulated on acid treatment until 18h. Thus, Sorbitoldehydrogenase and phosphofructo kinasegenes have a crucial role in diapause termination as evidenced by HCl treatment, while the other genes did not have major roles.

  14. Molecular cloning and characterization of a putative cDNA encoding endoglucanase IV from Trichoderma viride and its expression in Bombyx mori.

    Science.gov (United States)

    Li, Xing-Hua; Zhang, Peng; Liang, Shuang; Zhou, Fang; Wang, Mei-Xian; Bhaskar, Roy; Malik, Firdose Ahmad; Niu, Yan-Shan; Miao, Yun-Gen

    2012-01-01

    The development of cellulase production technology has greatly contributed to the successful use of cellulosic materials as renewable carbon sources. In this study, a putative endoglucanase IV (EG IV) complementary DNA was cloned from the mycelium of a strain of the filamentous fungus Trichoderma viride using a PCR-based exon-splicing method and expressed in both a silkworm BmN cell line and in silkworm larvae. Western blot analysis detected a band of 42 kDa in BmN cells after infection with a recombinant mBacmid/BmNPV/EG IV baculovirus. Sequence alignment analysis of the T. viride EG IV gene showed two domains that were highly conserved with glycosyl hydrolases and a funga-type cellulose-binding domain. Analysis of variance showed that silkworms infected with recombinant baculoviruses exhibited significantly higher enzyme activity that was 48.84% higher than silkworms infected with blank baculoviruses and 46.61% higher than normal silkworms. The expressed bioactive EG IV was also stable at the pH range from 5.0 to 10.0. The availability of large quantities of bioactive EG IV in silkworm provided a possibility to produce cellulase transgenic silkworm, which express bioactive cellulase specially in its digestive tract and improve its metabolism efficiency of mulberry leaves. Its application in the sericulture industry may be very promising.

  15. Expression in Escherichia coli and purification of bioactive antibacterial peptide ABP-CM4 from the Chinese silk worm, Bombyx mori.

    Science.gov (United States)

    Li, Bao-Cun; Zhang, Shuang-Quan; Dan, Wen-Bing; Chen, Yu-Qing; Cao, Peng

    2007-07-01

    The antibacterial peptide CM4 (ABP-CM4), isolated from Chinese Bombys mori, is a 35-residue cationic, amphipathic alpha-helical peptide that exhibits a broad range of antimicrobial activity. To explore a new approach for the expression of ABP-CM4 in E. coli, the gene ABP-CM4, obtained by recursive PCR (rPCR), was cloned into the vector pET32a to construct a fusion expression plasmid. The fusion protein Trx-CM4 was expressed in soluble form, purified by Ni(2+)-chelating chromatography, and cleaved by formic acid to release recombinant CM4. Purification of rCM4 was achieved by affinity chromatography and reverse-phase HPLC. The purified of recombinant peptide showed antimicrobial activities against E. coli K(12)D(31), Penicillium chrysogenum, Aspergillus niger and Gibberella saubinetii. According to the antimicrobial peptide database (http://aps.unmc.edu/AP/main.html), 116 peptides contain a Met residue, but only 5 peptides contain the AspPro site, indicating a broader application of formic acid than CNBr in cleaving fusion protein. The successful application to the expression of the ABP-CM4 indicates that the system is a low-cost, efficient way of producting milligram quantities of ABP-CM4 that is biologically active.

  16. Contribution to the ultrastructural study of silk-excretion cells and autoradiographic analysis of intracellular fibroin transport in Bombyx mori L

    International Nuclear Information System (INIS)

    Couble, Pierre.

    1974-01-01

    It is much easier to study the mechanisms involved in the synthesis and exportation of extracellular proteins in the biological material chosen is highly differentiated. The silk-excretion gland of the silkworm is ideal in this respect because during the larva period, especially at the end of the 5th and last stage, the cells at the rear (excreting tube) synthesize and export massive quantities of a single protein: fibroin. These phenomena were explored by a cytological study carried out mainly by electron microscopy and autoradiography. The results obtained are given. They relate first of all to the morphological development of the secretion tube cells from the end of the 4th larva stage to the spinning of the cocoon, and contribute new information on the cell changes during the 4th slough and the end of the 5th age. They also concern intracellular fibroin transport which is proved to take place through the Golgi apparatus, and finally the possible role of the microtubules and microfilaments in fibroin transport and secretion. On this last point the results so far constitute only, a preliminary approach which justifie no final conclusions; they merely suggest that the microfilaments of the apical region are involved in the secretion process [fr

  17. Desenvolvimento biológico das raças puras e dos híbridos do bicho-da-seda (Bombyx mori L., após o 5º ínstar, em duas estações do ano Biological development after 5th instar among pure breed and hybrids of silkworm (Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    Roque Takahashi

    2002-04-01

    Full Text Available O objetivo deste experimento foi o de verificar a duração da subida no bosque e formação do casulo; duração da formação do casulo até a emergência da mariposa e longevidade do adulto, assim como o peso médio do casulo verde e teor líquido de seda, entre duas raças puras - AJ1 (Japonesa e BC1 (Chinesa e dois híbridos, originários do cruzamento recíproco entre duas raças puras - BC1 X AJ1 (H1 e AJ1 X BC1 (H2, em duas épocas distintas: primavera (E1 e outono (E2. Os híbridos e BC1 foram superiores à raça pura AJ1 ao confeccionarem os casulos, levando em média 3,4 dias e 4,2 dias, respectivamente (E1. As lagartas criadas na E2, mostraram-se superiores em 77,88% em seus tempos de subida no bosque e confecção do casulo, em relação a E1. Quanto ao parâmetro emergência do adulto, sobressaiu-se o H2 (média de 8,9 dias, na E1, em relação às raças puras. As fêmeas dos híbridos, sobreviveram uma média de 3,8 dias a mais que as raças puras, para a E1. Na E2, o H1 destacou-se em relação às raças puras, cuja longevidade foi superior em 9,6 dias. Para os machos, na E2, a longevidade do H2 (5,0 dias foi superior à raça pura AJ1 (4,0 dias. Os híbridos e a AJ1 apresentaram casulos mais pesados em 15,2% em relação a BC1 (E2 e teor líquido de seda não foi significativo em ambas as épocas.This research was carried out to verify the duration of the rising up of caterpillar and the cocoon formation; duration of the cocoon formation to the emergency of the moth and the life span of the adult, as well as the medium weight of the green cocoon and liquid rate of silk, among two pure breeds - AJ1 (Japanese and BC1 (Chinese and two hybrids, from reciprocal crossing among two pure breeds - BC1 X AJ1 (H1 and AJ1 X BC1 (H2, in two different seasons: spring (S1 and autumn (S2. The hybrids and BC1 breed were superior to the pure race AJ1 making the cocoons, taking on average 3.4 days and 4.2 days, respectively (S1. The caterpillars grew in S2 were superior in 77.88% in their times of rising up and making the cocoon, in relation to S1. The hybrid H2 was better than the pure breeds in emergency of adult with average of 8.9 days, in S1. The females of the hybrid, survived an average of 3.8 more days than the pure breeds, in S1. In S2, H1 stood out in relation to the pure races, whose life span was superior in 9.6 days. For the males, in S2, the life span of H2 was 5.0 days. It was superior to the pure race AJ1 - 4.0 days. The hybrids and AJ1 presented heavier cocoons in 15.2% in relation to BC1 (S2 and liquid rate of silk was not significant in both seasons.

  18. Identification of 2nd chromosome region translocated onto the W chromosome by RFLP with EST-cDNA clones in the Gensei-kouken strains of the mulberry silkworm, Bombyx mori L

    Directory of Open Access Journals (Sweden)

    Sivaramakurup Sreekumar

    2010-01-01

    Full Text Available In silkworms, sex-limited strains are either obtained spontaneously or induced by X-rays or gamma rays. When a fragment of an autosome carrying a dominant allele of those genes responsible for certain characters is translocated onto a W chromosome, the female of the successive generations will express these phenotypic characters and sex discrimination can be facilitated. Gensei-kouken strains are sex-limited strains of silkworms developed by irradiating the pupae with gamma rays, by which a portion of the second chromosome is translocated onto the W chromosome. In these improved strains, the females are yellow-blooded and spin yellow cocoons. By using the EST-cDNA clones mapped on the Z chromosome, we identified the sex according to the polymorphic banding pattern or intensity of the signals. Furthermore, by using the clones on the second chromosome, the region of the second chromosome translocated onto the W chromosome was also defined. In both the A95 and A 96 strains selected for the present study, only the mid-portion of the second chromosome was translocated. The differences in length of the fragments translocated in these strains are discussed.

  19. EST Table: AV404642 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV404642 prgv0279 10/09/28 81 %/116 aa ref|NP_001091796.1| programmed cell death pr...otein 5-like protein [Bombyx mori] gb|ABJ97180.1| programmed cell death protein 5-like protein [Bombyx mori

  20. NCBI nr-aa BLAST: CBRC-AGAM-04-0130 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-AGAM-04-0130 ref|NP_001040188.1| KDEL endoplasmic reticulum protein retention ...receptor 2a [Bombyx mori] gb|ABD36213.1| KDEL endoplasmic reticulum protein retention receptor 2a [Bombyx mori] NP_001040188.1 3e-45 47% ...

  1. Cloning and characterization of peptidylprolyl isomerase B in the ...

    African Journals Online (AJOL)

    Peptidylprolyl isomerases (PPIases) play essential roles in protein folding and are implicated in immune response and cell cycle control. Our previous proteomic analysis indicated that Bombyx mori PPIases may be involved in anti- Bombyx mori nucleopolyhedrovirus (BmNPV) response. To help investigate this mechanism, ...

  2. EST Table: BY930925 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available 8 %/188 aa ref|NP_001116821.1| 18 wheeler [Bombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/08/29 64...gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] BY930925 ovS0 ...

  3. EST Table: CK516272 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available 9/29 61 %/212 aa ref|NP_001116821.1| 18 wheeler [Bombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/08...5 %/217 aa gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] CK516272 swj ...

  4. EST Table: FS756340 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available 7 %/154 aa ref|NP_001116821.1| 18 wheeler [Bombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/09/08 44...gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] FS756340 fcaL ...

  5. EST Table: CN375203 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CN375203 rzhswab0_003107 10/09/28 100 %/199 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/09/01 low homology 10/08/28

  6. EST Table: CK490566 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK490566 rswab0_008829.y1 10/09/29 98 %/198 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 low homology 10/08/28

  7. EST Table: BB985376 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985376 E_ET_MSV3_04C09_F_0 10/09/28 93 %/253 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 low homology 10/08/

  8. EST Table: BB985987 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985987 E_ET_MSV3_12C07_F_0 10/09/28 91 %/197 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 low homology 10/08/

  9. EST Table: CN375202 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CN375202 rzhswab0_006736 10/09/28 98 %/193 aa ref|NP_001166288.1| sericin 2 isoform... 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/09/01 low homology 10/08/28 l

  10. EST Table: BB986887 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986887 E_ET_MSV3_23E01_F_0 10/09/28 100 %/176 aa ref|NP_001166287.1| sericin 2 is...oform 1 precursor [Bombyx mori] gb|ADA84147.1| sericin 2 precursor [Bombyx mori] 10/08/28 low homology 10/08

  11. EST Table: CK486070 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK486070 rswab0_002530.y1 10/09/29 99 %/196 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 low homology 10/08/28

  12. EST Table: CK485783 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK485783 rswab0_002015.y1 10/09/29 100 %/165 aa ref|NP_001166288.1| sericin 2 isofo...rm 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 low homology 10/08/28

  13. EST Table: CK487117 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK487117 rswab0_004189.y1 10/09/29 100 %/207 aa ref|NP_001166288.1| sericin 2 isofo...rm 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 low homology 10/08/28

  14. EST Table: CK542413 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK542413 rswhb0_006901.y1 10/09/29 99 %/209 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/31 low homology 10/08/28

  15. EST Table: BB986088 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986088 E_ET_MSV3_13D10_F_0 10/09/28 100 %/193 aa ref|NP_001166287.1| sericin 2 is...oform 1 precursor [Bombyx mori] gb|ADA84147.1| sericin 2 precursor [Bombyx mori] 10/08/28 low homology 10/08

  16. EST Table: NM_001172816 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available NM_001172816 Ser2 10/09/29 87 %/1722 aa ref|NP_001166287.1| sericin 2 isoform 1 pre...cursor [Bombyx mori] gb|ADA84147.1| sericin 2 precursor [Bombyx mori] 10/09/13 low homology 10/08/29 low hom

  17. EST Table: CK539765 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK539765 rswhb0_002668.y1 10/09/29 92 %/221 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/31 low homology 10/08/28

  18. EST Table: CK485000 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK485000 rswab0_000812.y1 10/09/29 97 %/177 aa ref|NP_001166287.1| sericin 2 isofor...m 1 precursor [Bombyx mori] gb|ADA84147.1| sericin 2 precursor [Bombyx mori] 10/08/30 low homology 10/08/28

  19. EST Table: NM_001172817 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available NM_001172817 Ser2 11/12/09 n.h 10/09/29 81 %/864 aa ref|NP_001166288.1| sericin 2 i...soform 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/09/13 low homology 10/0

  20. EST Table: CK491362 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK491362 rswab0_009813.y1 10/09/29 95 %/172 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 low homology 10/08/28

  1. EST Table: BB985303 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985303 E_ET_MSV3_03D09_F_0 10/09/28 99 %/251 aa ref|NP_001166287.1| sericin 2 iso...form 1 precursor [Bombyx mori] gb|ADA84147.1| sericin 2 precursor [Bombyx mori] 10/08/28 low homology 10/08/

  2. EST Table: CK544252 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK544252 rswhb0_013739.y1 10/09/29 99 %/137 aa ref|NP_001040402.1| preimplantation ...protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/08/31 86 %/138 aa FBpp0116991|

  3. EST Table: FS781715 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS781715 E_FL_fcaL_38G13_R_0 10/09/28 97 %/209 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/08 80 %/210 aa FBpp02771

  4. EST Table: FS868421 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS868421 E_FL_fner_36N05_R_0 10/09/28 98 %/197 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/11 81 %/198 aa FBpp02771

  5. EST Table: FS763745 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS763745 E_FL_fcaL_34J22_F_0 10/09/28 98 %/202 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/08 83 %/203 aa FBpp02522

  6. EST Table: BW999096 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BW999096 SES2266 10/09/28 85 %/193 aa ref|NP_001040402.1| preimplantation protein [...Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/08/29 73 %/195 aa FBpp0252287|DwilGK231

  7. EST Table: DC543963 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available DC543963 E_FL_dpe-_23N04_F_0 10/09/28 100 %/178 aa ref|NP_001040402.1| preimplantat...ion protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/02 87 %/178 aa FBpp0252

  8. EST Table: FS780579 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS780579 E_FL_fcaL_34J22_R_0 10/09/28 98 %/202 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/08 80 %/203 aa FBpp02771

  9. EST Table: FS770331 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS770331 E_FL_fcaL_18O17_F_0 10/09/28 99 %/155 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/08 86 %/156 aa FBpp02522

  10. EST Table: FS909348 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS909348 E_FL_fufe_16P20_F_0 10/09/28 99 %/220 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/12 80 %/222 aa FBpp02346

  11. EST Table: NM_001046937 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available NM_001046937 LOC732938 10/09/29 100 %/220 aa ref|NP_001040402.1| preimplantation pr...otein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/13 79 %/222 aa FBpp0234606|Dv

  12. EST Table: FY756405 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY756405 E_FL_famL_20O01_R_0 11/11/04 97 %/196 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 11/11/04 80 %/197 aa FBpp02771

  13. EST Table: FS853020 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS853020 E_FL_fner_45D02_F_0 10/09/28 99 %/180 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/11 87 %/181 aa FBpp02522

  14. EST Table: FY000760 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY000760 bmov3c12 11/11/04 100 %/177 aa ref|NP_001040402.1| preimplantation protein... [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 11/11/04 87 %/178 aa FBpp0252287|DwilGK2

  15. EST Table: BP182279 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BP182279 NRPG0386 10/09/28 100 %/161 aa ref|NP_001040402.1| preimplantation protein... [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/08/29 87 %/162 aa FBpp0252287|DwilGK2

  16. EST Table: FS769451 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS769451 E_FL_fcaL_52D16_F_0 10/09/28 96 %/208 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/08 78 %/209 aa FBpp02346

  17. EST Table: CK511452 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK511452 rswdd0_007663.y1 10/09/29 100 %/132 aa ref|NP_001040402.1| preimplantation... protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/08/30 87 %/133 aa FBpp0116991

  18. EST Table: FY753429 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY753429 E_FL_famL_09K13_R_0 11/11/04 98 %/146 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 11/11/04 80 %/147 aa FBpp02771

  19. EST Table: CK561354 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK561354 rswpb0_003859.y1 10/09/29 100 %/136 aa ref|NP_001040402.1| preimplantation... protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/01 87 %/137 aa FBpp0116991

  20. EST Table: FS917608 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS917608 E_FL_fufe_41N17_F_0 10/09/28 99 %/220 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/12 80 %/222 aa FBpp02346

  1. EST Table: FS841606 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS841606 E_FL_fner_12N15_F_0 10/09/28 100 %/132 aa ref|NP_001040402.1| preimplantat...ion protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/10 87 %/133 aa FBpp0116

  2. EST Table: FS876378 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS876378 E_FL_ftes_06K04_F_0 10/09/28 100 %/154 aa ref|NP_001040402.1| preimplantat...ion protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/11 87 %/155 aa FBpp0252

  3. EST Table: CK534545 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK534545 rswgb0_002719.y1 10/09/29 100 %/146 aa ref|NP_001040402.1| preimplantation... protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/08/31 87 %/147 aa FBpp0116991

  4. EST Table: CN211963 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CN211963 rzhswbb0_006380 10/09/28 100 %/136 aa ref|NP_001040402.1| preimplantation ...protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/01 87 %/137 aa FBpp0116991|

  5. EST Table: FS871345 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS871345 E_FL_fner_45D02_R_0 10/09/28 99 %/220 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/11 80 %/221 aa FBpp02346

  6. EST Table: FY011485 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY011485 rbmov3c12 11/11/04 98 %/164 aa ref|NP_001040402.1| preimplantation protein... [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 11/11/04 78 %/169 aa FBpp0234606|DvirGJ2

  7. EST Table: CK537808 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK537808 rswgb0_007090.y1 10/09/29 97 %/192 aa ref|NP_001040402.1| preimplantation ...protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/08/31 80 %/192 aa FBpp0252287|

  8. EST Table: BY941180 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BY941180 E_FL_e100_12L15_F_0 10/09/28 97 %/139 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/08/30 85 %/140 aa FBpp01169

  9. EST Table: FS743516 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS743516 E_FL_bmmt_25O09_R_0 10/09/28 98 %/167 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/07 80 %/167 aa FBpp02771

  10. EST Table: FS785832 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS785832 E_FL_fcaL_52D16_R_0 10/09/28 97 %/118 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/09 78 %/119 aa FBpp02346

  11. EST Table: CK541363 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK541363 rswhb0_005327.y1 10/09/29 100 %/113 aa ref|NP_001040402.1| preimplantation... protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/08/31 85 %/114 aa FBpp0116991

  12. EST Table: CK544721 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK544721 rswhb0_014958.y1 10/09/29 100 %/142 aa ref|NP_001040402.1| preimplantation... protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/08/31 88 %/143 aa FBpp0116991

  13. EST Table: FS764958 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS764958 E_FL_fcaL_38G13_F_0 10/09/28 98 %/220 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/08 80 %/222 aa FBpp02346

  14. EST Table: FY008863 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY008863 bmov25n20 11/11/04 99 %/180 aa ref|NP_001040402.1| preimplantation protein... [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 11/11/04 87 %/181 aa FBpp0252287|DwilGK2

  15. EST Table: FS917079 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS917079 E_FL_fufe_40D23_F_0 10/09/28 99 %/217 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/12 81 %/213 aa FBpp02346

  16. EST Table: FS909374 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS909374 E_FL_fufe_17B02_F_0 10/09/28 99 %/220 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/12 80 %/222 aa FBpp02346

  17. EST Table: FS732209 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS732209 E_FL_bmmt_25O09_F_0 10/09/28 100 %/175 aa ref|NP_001040402.1| preimplantat...ion protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/03 88 %/176 aa FBpp0252

  18. EST Table: CK559669 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK559669 rswpb0_001761.y1 10/09/29 98 %/103 aa ref|NP_001040402.1| preimplantation ...protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/01 81 %/104 aa FBpp0116991|

  19. EST Table: FS931382 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS931382 E_FL_fwgP_31A18_F_0 10/09/28 100 %/158 aa ref|NP_001040402.1| preimplantat...ion protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/13 87 %/159 aa FBpp0252

  20. EST Table: CK559584 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK559584 rswpb0_001653.y1 10/09/29 100 %/156 aa ref|NP_001040402.1| preimplantation... protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/01 87 %/157 aa FBpp0252287

  1. EST Table: FY743324 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY743324 E_FL_famL_20O01_F_0 11/11/04 98 %/207 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 11/11/04 86 %/197 aa FBpp02522

  2. EST Table: FS912488 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS912488 E_FL_fufe_26G13_F_0 10/09/28 99 %/220 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/12 80 %/222 aa FBpp02346

  3. EST Table: FS910496 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS910496 E_FL_fufe_20G12_F_0 10/09/28 99 %/220 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/12 80 %/222 aa FBpp02346

  4. EST Table: CK521898 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK521898 rswea0_006921.y1 10/09/29 98 %/140 aa ref|NP_001040402.1| preimplantation ...protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/08/31 86 %/141 aa FBpp0116991|

  5. EST Table: FS786649 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS786649 E_FL_fcaL_18O17_R_0 10/09/28 98 %/167 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/09 80 %/167 aa FBpp02771

  6. EST Table: FS906090 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS906090 E_FL_fufe_07B18_F_0 10/09/28 99 %/220 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/12 80 %/222 aa FBpp02346

  7. EST Table: FS850077 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS850077 E_FL_fner_36N05_F_0 10/09/28 99 %/204 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/11 83 %/205 aa FBpp02522

  8. EST Table: FS878889 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS878889 E_FL_ftes_13O22_F_0 10/09/28 96 %/166 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/11 85 %/167 aa FBpp02522

  9. EST Table: FS860309 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS860309 E_FL_fner_12N15_R_0 10/09/28 98 %/165 aa ref|NP_001040402.1| preimplantati...on protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/11 80 %/166 aa FBpp02771

  10. EST Table: CK560024 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK560024 rswpb0_002234.y1 10/09/29 97 %/172 aa ref|NP_001040402.1| preimplantation ...protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/09/01 87 %/173 aa FBpp0252287|

  11. EST Table: FY019699 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY019699 rbmov25n20 11/11/04 98 %/168 aa ref|NP_001040402.1| preimplantation protei...n [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 11/11/04 79 %/169 aa FBpp0277152|DpseGA

  12. EST Table: CK488500 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK488500 rswab0_006077.y1 10/09/29 97 %/137 aa ref|NP_001040402.1| preimplantation ...protein [Bombyx mori] gb|ABF51322.1| preimplantation protein [Bombyx mori] 10/08/30 85 %/138 aa FBpp0116991|

  13. EST Table: FS871051 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available aa ref|NP_001139536.1| mortality factor 4-like [Bombyx mori] gb|ABJ99463.1| mrg15-like protein [Bombyx mori...FS871051 E_FL_fner_44F14_R_0 11/12/09 GO hit GO:0005634(nucleus) 10/09/28 100 %/257

  14. EST Table: BY930853 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ecursor dbj|BAA06160.1| humoral lectin prepropeptide [Bombyx mori] prf||2106140A hemocytin 10/08/29 low homo...cursor [Bombyx mori] sp|P98092.1|HMCT_BOMMO RecName: Full=Hemocytin; AltName: Full=Humoral lectin; Flags: Pr

  15. EST Table: DC551726 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ecursor dbj|BAA06160.1| humoral lectin prepropeptide [Bombyx mori] prf||2106140A hemocytin 10/09/02 31 %/251...cursor [Bombyx mori] sp|P98092.1|HMCT_BOMMO RecName: Full=Hemocytin; AltName: Full=Humoral lectin; Flags: Pr

  16. Author Details

    African Journals Online (AJOL)

    Abdel-Nabi, IM. Vol 10 (2008) - Articles Biological and technological effects of mulberry varieties and nutritional additives on silkworm Bombyx mori development. Abstract · Vol 10 (2008) - Articles The effect of mulberry varieties and nutritional additives on the protein patterns of the silkworm Bombyx mori. Abstract.

  17. EST Table: FS839797 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS839797 E_FL_fner_07L06_F_0 10/09/28 100 %/142 aa ref|NP_001037402.1| arginine kin...ase [Bombyx mori] gb|ABB88514.1| arginine kinase [Bombyx mori] gb|ABD36282.1| arginine kinase [Bombyx mori] gb|ABY597...rotein|UNKN:39405743:39406543:-1|gene:AGAP012924 10/09/10 83 %/142 aa gnl|Amel|GB10973-PA 10/09/10 84 %/146 aa gi|27001497

  18. Expression of polyhedrin-hEGF fusion protein in cultured cells and ...

    African Journals Online (AJOL)

    GRACE

    2006-06-02

    , protease K, X-gal and related reagents were purchased from Roche ..... fusion protein in silkworm larvae infected with recombinant Bombyx mori nuclear polyhedrosis virus. J. Gen. Virol. 68: 2599-2606. Massotte D (2003).

  19. Short hairpin RNA expression for enhancing the resistance of ...

    African Journals Online (AJOL)

    Short hairpin RNA expression for enhancing the resistance of Bombyx mori (Bm) to nucleopolyhedrovirus in vitro and in vivo. Roy Bhaskar, Fang Zhou, Shuang Liang, Wan-Fu Yue, Yan-shan Niu, Yun- gen Miao ...

  20. Purification and biochemical characterization of a novel glutathione ...

    African Journals Online (AJOL)

    SERVER

    2008-02-05

    dinitrobenzene as a substrate. ..... purification of GST enzyme from other insect species, which were from 3 to 26% in Hyphantria .... Glutathione S-transferases from the larval gut of the silkworm. Bombyx mori: cDNA cloning, gene ...

  1. EST Table: BW998041 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BW998041 ES3138 10/09/28 54 %/127 aa ref|NP_001165388.1| aliphatic nitrilase [Bomby...x mori] gb|ADB27116.1| aliphatic nitrilase [Bombyx mori] 10/08/29 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS789730 L8 ...

  2. EST Table: BY941476 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BY941476 E_FL_e100_14F06_F_0 10/09/28 100 %/109 aa ref|NP_001116821.1| 18 wheeler [...Bombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 low homology FS922922 e100 ...

  3. EST Table: DC540075 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available DC540075 E_FL_dpe-_04D16_F_0 10/09/28 97 %/176 aa ref|NP_001098700.1| nanos-M [Bomb...yx mori] dbj|BAF73619.1| nanos-M [Bombyx mori] 10/09/02 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 low homology 10/09/10 n.h FS918893 dpe- ...

  4. EST Table: CK562369 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK562369 rswpb0_005348.y1 10/09/29 100 %/182 aa ref|NP_001098700.1| nanos-M [Bombyx... mori] dbj|BAF73619.1| nanos-M [Bombyx mori] 10/09/01 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 low homology 10/09/10 n.h FS918893 swp ...

  5. EST Table: FY001942 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY001942 bmov6h18 11/11/04 100 %/147 aa ref|NP_001098702.1| nanos-like protein [Bom...byx mori] gb|ABS17681.1| nanos-like protein [Bombyx mori] 11/11/04 n.h 11/11/04 n.h 11/11/04 n.h 11/11/04 n.h 11/11/04 n.h NM_001105232 bmov ...

  6. EST Table: DC541400 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available DC541400 E_FL_dpe-_11C20_F_0 10/09/28 97 %/168 aa ref|NP_001098700.1| nanos-M [Bomb...yx mori] dbj|BAF73619.1| nanos-M [Bombyx mori] 10/09/02 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS918893 dpe- ...

  7. EST Table: AV403457 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403457 msgV0503 10/09/28 90 %/177 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  8. EST Table: CK488480 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK488480 rswab0_006045.y1 10/09/29 86 %/145 aa ref|NP_001166287.1| sericin 2 isofor...m 1 precursor [Bombyx mori] gb|ADA84147.1| sericin 2 precursor [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 swa ...

  9. EST Table: CK490794 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK490794 rswab0_009111.y1 10/09/29 95 %/163 aa ref|NP_001037506.1| sericin 1 [Bomby...x mori] emb|CAA88741.1| sericin1B [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB986107 swa ...

  10. EST Table: CK490249 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK490249 rswab0_008252.y1 10/09/29 92 %/213 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 low homology 10/09/10 n.h 10/09/10 n.h NM_001172817 swa ...

  11. EST Table: AV403247 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403247 msgV0234 10/09/28 91 %/189 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  12. EST Table: AV403467 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403467 msgV0519 10/09/28 85 %/159 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  13. EST Table: CK488539 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK488539 rswab0_006125.y1 10/09/29 91 %/202 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 swa ...

  14. EST Table: AV403356 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403356 msgV0366 10/09/28 66 %/121 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 msgV ...

  15. EST Table: AV403304 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403304 msgV0310 10/09/28 85 %/165 aa ref|NP_001166287.1| sericin 2 isoform 1 prec...ursor [Bombyx mori] gb|ADA84147.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  16. EST Table: AV403504 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403504 msgV0623 10/09/28 98 %/114 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  17. EST Table: AV403487 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403487 msgV0602 10/09/28 95 %/130 aa ref|NP_001166287.1| sericin 2 isoform 1 prec...ursor [Bombyx mori] gb|ADA84147.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  18. EST Table: CN212174 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CN212174 rzhswab0_007010 10/09/28 77 %/192 aa ref|NP_001166288.1| sericin 2 isoform... 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/09/01 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 L12 ...

  19. EST Table: AV403164 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403164 msgV0119 10/09/28 84 %/120 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  20. EST Table: AV403698 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403698 msgV0982 10/09/28 85 %/164 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  1. EST Table: AV403268 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403268 msgV0258 10/09/28 62 %/158 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  2. EST Table: CK485971 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK485971 rswab0_002234.y1 10/09/29 85 %/164 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 swa ...

  3. EST Table: CK485820 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK485820 rswab0_002053.y1 10/09/29 91 %/195 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 swa ...

  4. EST Table: CK490530 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK490530 rswab0_008786.y1 10/09/29 85 %/167 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 swa ...

  5. EST Table: CK491617 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK491617 rswab0_010162.y1 10/09/29 90 %/188 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 swa ...

  6. EST Table: AV403374 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403374 msgV0392 10/09/28 86 %/135 aa ref|NP_001166287.1| sericin 2 isoform 1 prec...ursor [Bombyx mori] gb|ADA84147.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  7. EST Table: AV403444 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403444 msgV0485 10/09/28 80 %/156 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  8. EST Table: AV403508 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403508 msgV0629 10/09/28 79 %/122 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 msgV ...

  9. EST Table: AV403123 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403123 msgV0067 10/09/28 65 %/125 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 msgV ...

  10. EST Table: AV403443 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403443 msgV0483 10/09/28 62 %/162 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 msgV ...

  11. EST Table: AV403596 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403596 msgV0735 10/09/28 60 %/200 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 msgV ...

  12. EST Table: CK488498 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK488498 rswab0_006074.y1 10/09/29 90 %/161 aa ref|NP_001166288.1| sericin 2 isofor...m 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 swa ...

  13. EST Table: Z48802 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Z48802 DC550742 11/12/09 n.h 10/09/29 80 %/583 aa ref|NP_001037506.1| sericin 1 [Bo...mbyx mori] emb|CAA88741.1| sericin1B [Bombyx mori] 10/09/13 low homology 10/08/29 low homology 10/09/10 n.h 10/09/10 n.h 10/09/10 low homology Z48802 ...

  14. EST Table: AV403709 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AV403709 msgV0994 10/09/28 61 %/173 aa ref|NP_001166288.1| sericin 2 isoform 2 prec...ursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 msgV ...

  15. EST Table: CK499683 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK499683 rswbb0_010208.y1 10/09/29 91 %/184 aa ref|NP_001091744.1| alkaline nucleas...e [Bombyx mori] dbj|BAF33251.1| alkaline nuclease [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS800660 swb ...

  16. EST Table: FY025038 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY025038 bmte10l11 11/11/04 37 %/142 aa ref|NP_001091744.1| alkaline nuclease [Bomb...yx mori] dbj|BAF33251.1| alkaline nuclease [Bombyx mori] 11/11/04 n.h 11/11/04 n.h 11/11/04 n.h 11/11/04 n.h 11/11/04 n.h FS882103 bmte ...

  17. EST Table: FS804780 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS804780 E_FL_fmgV_16C05_F_0 11/12/09 n.h 10/09/28 99 %/218 aa ref|NP_001124364.1| tachykinin... [Bombyx mori] dbj|BAG50368.1| tachykinin [Bombyx mori] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/...09/10 n.h 10/09/10 30 %/221 aa gi|189240200|ref|XP_975364.2| PREDICTED: similar to preprotachykinin [Tribolium castaneum] FS804780 fmgV ...

  18. EST Table: BY940884 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BY940884 E_FL_e100_11D16_F_0 10/09/28 96 %/114 aa ref|NP_001116821.1| 18 wheeler [B...ombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 low homology 10/09/1...0 n.h 10/09/10 38 %/100 aa gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] FS922922 e100 ...

  19. EST Table: DC555421 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available DC555421 E_FL_wd--_10C09_F_0 10/09/28 100 %/113 aa ref|NP_001116821.1| 18 wheeler [...Bombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/09/02 n.h 10/08/28 n.h 10/09/10 low homology 10/09/...10 n.h 10/09/10 37 %/100 aa gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] FS922922 wd-- ...

  20. EST Table: FY749160 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY749160 E_FL_famL_41E18_F_0 11/11/04 53 %/237 aa ref|NP_001116821.1| 18 wheeler [B...ombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 11/11/04 38 %/259 aa FBpp0156927|DgriGH23021-PA 11/11/0...11/11/04 36 %/253 aa gnl|Amel|GB15177-PA 11/11/04 43 %/262 aa gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] BY938801 famL ...

  1. EST Table: DC553638 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available DC553638 E_FL_wd--_04N17_F_0 10/09/28 100 %/165 aa ref|NP_001116821.1| 18 wheeler [...Bombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/09/02 61 %/165 aa FBpp0278316|DpseGA21191-PA 10/08/...7 10/09/10 64 %/165 aa gnl|Amel|GB15177-PA 10/09/10 72 %/165 aa gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] FS922922 wd-- ...

  2. EST Table: FY761239 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY761239 E_FL_famL_41E18_R_0 11/11/04 58 %/227 aa ref|NP_001116821.1| 18 wheeler [B...ombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 11/11/04 37 %/161 aa FBpp0278316|DpseGA21191-PA 11/11/0...a gnl|Amel|GB15177-PA 11/11/04 55 %/137 aa gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] BY943188 famL ...

  3. EST Table: FS755467 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS755467 E_ET_caL-_19O13_R_0 10/09/28 87 %/114 aa ref|NP_001116821.1| 18 wheeler [B...ombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/09/08 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BY944435 caL- ...

  4. EST Table: FS929627 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS929627 E_FL_fwgP_25L02_F_0 10/09/28 62 %/258 aa ref|NP_001116821.1| 18 wheeler [B...ombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/09/13 38 %/278 aa FBpp0245036|DwilGK15893-PA 10/08/2...Amel|GB15177-PA 10/09/10 51 %/263 aa gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] CK516272 fwgP ...

  5. EST Table: FS922922 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS922922 E_FL_fwgP_06A21_F_0 11/12/09 n.h 10/09/28 82 %/176 aa ref|NP_001116821.1| 18 wheel...er [Bombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/09/13 n.h 10/08/29 n.h 10/09/10 n.h 10/...09/10 n.h 10/09/10 36 %/129 aa gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] FS922922 fwgP ...

  6. EST Table: DC553735 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available DC553735 E_FL_wd--_05C09_F_0 10/09/28 93 %/206 aa ref|NP_001116821.1| 18 wheeler [B...ombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/09/02 44 %/224 aa FBpp0257093|DyakGE12083-PA 10/08/2...05 aa gnl|Amel|GB15177-PA 10/09/10 57 %/204 aa gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] FS922922 wd-- ...

  7. EST Table: FS749709 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS749709 E_ET_caL-_19O13_F_0 10/09/28 92 %/228 aa ref|NP_001116821.1| 18 wheeler [B...ombyx mori] dbj|BAB85498.1| 18 wheeler [Bombyx mori] 10/09/08 47 %/148 aa FBpp0235352|DvirGJ20935-PA 10/08/2...Amel|GB15177-PA 10/09/10 51 %/215 aa gi|91076478|ref|XP_972409.1| PREDICTED: similar to 18 wheeler [Tribolium castaneum] FS922922 caL- ...

  8. EST Table: FS813939 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS813939 E_FL_fmgV_41L05_F_0 10/09/28 65 %/121 aa ref|NP_001136225.1| hypothetical ...protein LOC100216500 [Bombyx mori] gb|ACJ66918.1| hypothetical protein 39 [Bombyx mori] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h AU001277 fmgV ...

  9. EST Table: BB986928 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986928 E_ET_MSV3_24A01_F_0 10/09/28 80 %/122 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 MSV3 ...

  10. EST Table: BB986509 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986509 E_ET_MSV3_18F09_F_0 10/09/28 85 %/176 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 MSV3 ...

  11. EST Table: BB985551 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985551 E_ET_MSV3_06E06_F_0 10/09/28 74 %/267 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 MSV3 ...

  12. EST Table: BB991701 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB991701 E_ET_MSV3_18B06_R_0 10/09/28 100 %/100 aa ref|NP_001166288.1| sericin 2 is...oform 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990706 MSV3 ...

  13. EST Table: BB992001 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB992001 E_ET_MSV3_23C09_R_0 11/12/09 n.h 10/09/28 89 %/141 aa ref|NP_001166288.1| sericin... 2 isoform 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB992001 MSV3 ...

  14. EST Table: BB987076 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB987076 E_ET_MSV3_26A12_F_0 10/09/28 80 %/210 aa ref|NP_001037506.1| sericin 1 [Bo...mbyx mori] emb|CAA88741.1| sericin1B [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h Z48802 MSV3 ...

  15. EST Table: BB990891 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB990891 E_ET_MSV3_04C09_R_0 10/09/28 89 %/142 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB992001 MSV3 ...

  16. EST Table: BB986444 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986444 E_ET_MSV3_17H04_F_0 10/09/28 76 %/126 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 MSV3 ...

  17. EST Table: BB986461 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986461 E_ET_MSV3_18A12_F_0 10/09/28 100 %/153 aa ref|NP_001166288.1| sericin 2 is...oform 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 low homology NM_001172817 MSV3 ...

  18. EST Table: BB985437 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985437 E_ET_MSV3_05B01_F_0 10/09/28 79 %/277 aa ref|NP_001037506.1| sericin 1 [Bo...mbyx mori] emb|CAA88741.1| sericin1B [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h Z48802 MSV3 ...

  19. EST Table: BB986645 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986645 E_ET_MSV3_20D04_F_0 10/09/28 100 %/152 aa ref|NP_001166288.1| sericin 2 is...oform 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 low homology 10/09/10 n.h 10/09/10 n.h NM_001172817 MSV3 ...

  20. EST Table: BB986838 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986838 E_ET_MSV3_22H07_F_0 10/09/28 92 %/223 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 low homology 10/08/27 low homology 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 MSV3 ...

  1. EST Table: BB986359 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986359 E_ET_MSV3_16F06_F_0 10/09/28 61 %/185 aa ref|NP_001037506.1| sericin 1 [Bo...mbyx mori] emb|CAA88741.1| sericin1B [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB986107 MSV3 ...

  2. EST Table: BB985569 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985569 E_ET_MSV3_06G02_F_0 11/12/09 n.h 10/09/28 73 %/265 aa ref|NP_001166288.1| sericin... 2 isoform 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 MSV3 ...

  3. EST Table: BB987079 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB987079 E_ET_MSV3_26B05_F_0 10/09/28 81 %/128 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 MSV3 ...

  4. EST Table: BB992045 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB992045 E_ET_MSV3_24A01_R_0 10/09/28 85 %/157 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990706 MSV3 ...

  5. EST Table: BB986467 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986467 E_ET_MSV3_18B06_F_0 10/09/28 90 %/174 aa ref|NP_001166287.1| sericin 2 iso...form 1 precursor [Bombyx mori] gb|ADA84147.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 MSV3 ...

  6. EST Table: BB985921 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985921 E_ET_MSV3_11E05_F_0 10/09/28 87 %/195 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 low homology 10/09/10 n.h 10/09/10 n.h NM_001172817 MSV3 ...

  7. EST Table: BB991212 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB991212 E_ET_MSV3_09G07_R_0 10/09/28 62 %/208 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990706 MSV3 ...

  8. EST Table: BB986491 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986491 E_ET_MSV3_18D09_F_0 10/09/28 90 %/185 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h NM_001172817 MSV3 ...

  9. EST Table: BB985711 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985711 E_ET_MSV3_08H08_F_0 10/09/28 81 %/238 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 MSV3 ...

  10. EST Table: BB991931 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB991931 E_ET_MSV3_22D03_R_0 10/09/28 89 %/139 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB992001 MSV3 ...

  11. EST Table: BB991303 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB991303 E_ET_MSV3_12A08_R_0 10/09/28 78 %/193 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990706 MSV3 ...

  12. EST Table: BB985779 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985779 E_ET_MSV3_09G07_F_0 10/09/28 76 %/185 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 MSV3 ...

  13. EST Table: BB991685 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB991685 E_ET_MSV3_17H04_R_0 10/09/28 91 %/187 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990706 MSV3 ...

  14. EST Table: BB992013 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB992013 E_ET_MSV3_23E01_R_0 10/09/28 86 %/109 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB992001 MSV3 ...

  15. EST Table: BB985678 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985678 E_ET_MSV3_08E02_F_0 10/09/28 78 %/258 aa ref|NP_001037506.1| sericin 1 [Bo...mbyx mori] emb|CAA88741.1| sericin1B [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h Z48802 MSV3 ...

  16. EST Table: BB990706 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB990706 E_ET_MSV3_01D08_R_0 11/12/09 n.h 10/09/28 83 %/200 aa ref|NP_001166287.1| sericin... 2 isoform 1 precursor [Bombyx mori] gb|ADA84147.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990706 MSV3 ...

  17. EST Table: BB985753 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985753 E_ET_MSV3_09D11_F_0 10/09/28 77 %/109 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 MSV3 ...

  18. EST Table: BB985966 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB985966 E_ET_MSV3_12A08_F_0 10/09/28 81 %/234 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB985569 MSV3 ...

  19. EST Table: BB986294 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB986294 E_ET_MSV3_15H07_F_0 10/09/28 75 %/221 aa ref|NP_001037506.1| sericin 1 [Bo...mbyx mori] emb|CAA88741.1| sericin1B [Bombyx mori] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB986107 MSV3 ...

  20. EST Table: BB991973 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB991973 E_ET_MSV3_22H07_R_0 10/09/28 89 %/139 aa ref|NP_001166288.1| sericin 2 iso...form 2 precursor [Bombyx mori] gb|ADB04958.1| sericin 2 precursor [Bombyx mori] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB992001 MSV3 ...