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Sample records for oak silkworm antheraea

  1. The origin and dispersal of the domesticated Chinese oak silkworm, Antheraea pernyi, in China: a reconstruction based on ancient texts.

    PubMed

    Liu, Yanqun; Li, Yuping; Li, Xisheng; Qin, Li

    2010-01-01

    Sericulture is one of the great inventions of the ancient Chinese. Besides the mulberry silkworm (Bombyx mori), Chinese farmers developed rearing of the Chinese oak silkworm (Antheraea pernyi) about 400 years ago. In this paper, the historic records of the origins and dispersal of the domesticated Chinese oak silkworm in China are summarized. The first document with clearly recorded oak silkworm artificial rearing appeared in 1651, although Chinese oak silkworm was documented in about 270 AD. All of the evidence in the available historic records suggests that the domesticated Chinese oak silkworm originated in central and southern areas of Shandong Province in China around the 16th century, and then was introduced directly and indirectly by human commerce into the present habitations in China after the late 17th century. The results strongly support the hypothesis that only one geographically distinct event occurred in domestication of the modern Chinese oak silkworm.

  2. Preferential codon usage and two types of repetitive motifs in the fibroin gene of the Chinese oak silkworm, Antheraea pernyi.

    PubMed

    Yukuhiro, K; Kanda, T; Tamura, T

    1997-02-01

    In this paper we describe the peculiar structures and preferential codon usage found in wild silkworm fibroin genes. We determined a 1350 bp nucleotide sequence from the Chinese oak silkworm, Antheraea pernyi. The deduced amino acid sequence was partitioned into thirteen polyalanine-containing repetitive motifs, which was one of the characteristics of Antheraea fibroins. Eleven of these arrays can be classified into two types of motifs depending on difference in amino acid sequences following polyalanine. Repetitive motifs structurally similar to those of A. pernyi were detected in a homologue of the Japanese oak silkworm, Antheraea yamamai. The most remarkable feature of this study was preferential codon usage, especially seen in alanine synonymous codons within both homologues of Antheraea: isocodon GCA most frequently occurred in alanine isocodons. In contrast, GCU isocodon was the most abundant in Bombyx mori fibroin heavy chain that lacks polyalanine arrays. This result strongly suggests different modes of selective constraint between the two types of fibroin gene. The similar finding that GCA isocodon was most frequent in two dragline silk sequences of the spider, Nephila clavipes, is consistent with our results because of the repetitive polyalanine-containing arrays seen in spider dragline silk.

  3. Identification and Characterization of a Novel Microvitellogenin from the Chinese Oak Silkworm Antheraea pernyi.

    PubMed

    Liu, Yanqun; Chen, Miaomiao; Su, Junfang; Ma, Hongfang; Zheng, Xixi; Li, Qun; Shi, Shenglin; Qin, Li

    2015-01-01

    Microvitellogenin (mVg) is a relatively small vitellogenic protein only characterized in the eggs of the lepidopteran insects Manduca sexta and Bombyx mori. In the present study, we report a novel mVg (ApmVg) isolated from the Chinese oak silkworm Antheraea pernyi. The obtained ApmVg cDNA sequence contains an open reading frame of 783 bp encoding a protein of 260 amino acids with a predicted molecular weight of 29.96 kDa. This gene does not contain introns. Structural analysis revealed that this protein shares putative conserved domains with the lepidopteran low-molecular weight lipoprotein, which belongs to the lipoprotein_11 superfamily. The protein sequence of ApmVg exhibits 48% sequence identity with mVg from M. sexta and 40-47% sequence identity with the 30K lipoproteins from B. mori. Phylogenetic analysis suggests that ApmVg is a novel member of the lepidopteran low-molecular weight lipoproteins. Transcriptional analysis indicated that ApmVg mRNA is mainly expressed in the fat body (both female and male) during post-diapause development of the pupal stage, and it was also detected in ovaries and spermaries in smaller amounts. RT-PCR and Western blot analyses revealed that ApmVg is synthesized by the fat body and secreted into hemolymph and ultimately accumulates in eggs. The ApmVg transcript can be detected in the fat bodies of female pupae four days after treatment with 20-hydroxyecdysone and shows an expression pattern distinct from that of vitellogenin (Vg), which is detectable throughout diapausing and in post-diapause development. ApmVg decreased dramatically during embryonic development. These results represent the first study of mVg outside M. sexta and B. mori and provide insight into the physiological role and evolution of mVgs. PMID:26126120

  4. The Toll Signaling Pathway in the Chinese Oak Silkworm, Antheraea pernyi: Innate Immune Responses to Different Microorganisms.

    PubMed

    Sun, Ying; Jiang, Yiren; Wang, Yong; Li, Xisheng; Yang, Ruisheng; Yu, Zhiguo; Qin, Li

    2016-01-01

    The Toll pathway is one of the most important signaling pathways regulating insect innate immunity. Spatzle is a key protein that functions as a Toll receptor ligand to trigger Toll-dependent expression of immunity-related genes. In this study, a novel spatzle gene (ApSPZ) from the Chinese oak silkworm Antheraea pernyi was identified. The ApSPZ cDNA is 1065 nucleotides with an open reading frame (ORF) of 777 bp encoding a protein of 258 amino acids. The protein has an estimated molecular weight of 29.71 kDa and an isoelectric point (PI) of 8.53. ApSPZ is a nuclear and secretory protein with no conserved domains or membrane helices and shares 40% amino acid identity with SPZ from Manduca sexta. Phylogenetic analysis indicated that ApSPZ might be a new member of the Spatzle type 1 family, which belongs to the Spatzle superfamily. The expression patterns of several genes involved in the Toll pathway were examined at different developmental stages and various tissues in 5th instar larvae. The examined targets included A. pernyi spatzle, GNBP, MyD88, Tolloid, cactus and dorsalA. The RT-PCR results showed that these genes were predominantly expressed in immune-responsive fat body tissue, indicating that the genes play a crucial role in A. pernyi innate immunity. Moreover, A. pernyi infection with the fungus Nosema pernyi and the gram-positive bacterium Enterococcus pernyi, but not the gram-negative bacterium Escherichia coli, activated the Toll signaling pathway. These results represent the first study of the Toll pathway in A. pernyi, which provides insight into the A. pernyi innate immune system. PMID:27483463

  5. Overexpression of small heat shock protein 21 protects the Chinese oak silkworm Antheraea pernyi against thermal stress.

    PubMed

    Liu, Qiu-Ning; Zhu, Bao-Jian; Dai, Li-Shang; Fu, Wei-Wei; Lin, Kun-Zhang; Liu, Chao-Liang

    2013-08-01

    Small heat shock proteins (sHSPs) usually act as molecular chaperones to prevent proteins from being denatured in extreme conditions. We first report the sHSP21 gene, named as Ap-sHSP21, in the Chinese oak silkworm Antheraea pernyi (Lepidoptera: Saturniidae). The full-length cDNA of Ap-sHSP21 is 976 bp, including a 5'-untranslated region (UTR) of 99 bp, a 3'-UTR of 316 bp and an open reading frame (ORF) of 561 bp encoding a polypeptide of 186 amino acids. The deduced A. pernyi sHSP21 protein sequence reveals the percent identity is 82-93% in comparison to other sHSPs from insects. Real-time quantitative reverse transcription-PCR (qRT-PCR) analysis shows that Ap-sHSP21 expression is higher in testis than that in other examined tissues and significantly up-regulated after heat shock. In addition, prokaryotic expression and purification of the Ap-sHSP21 protein were performed. SDS-PAGE and Western blot analysis demonstrated that a 25 kDa recombinant protein was successfully expressed in Escherichia coli cells and the purified recombinant protein was also confirmed to protect restriction enzymes from thermal inactivation. The expression of Ap-sHSP21 was significantly down-regulated after RNA interference, which was confirmed by qRT-PCR and Western blot analysis. All together, these results suggest that Ap-sHSP21 play a key role in thermal tolerance.

  6. The Toll Signaling Pathway in the Chinese Oak Silkworm, Antheraea pernyi: Innate Immune Responses to Different Microorganisms

    PubMed Central

    Wang, Yong; Li, Xisheng; Yang, Ruisheng; Yu, Zhiguo; Qin, Li

    2016-01-01

    The Toll pathway is one of the most important signaling pathways regulating insect innate immunity. Spatzle is a key protein that functions as a Toll receptor ligand to trigger Toll-dependent expression of immunity-related genes. In this study, a novel spatzle gene (ApSPZ) from the Chinese oak silkworm Antheraea pernyi was identified. The ApSPZ cDNA is 1065 nucleotides with an open reading frame (ORF) of 777 bp encoding a protein of 258 amino acids. The protein has an estimated molecular weight of 29.71 kDa and an isoelectric point (PI) of 8.53. ApSPZ is a nuclear and secretory protein with no conserved domains or membrane helices and shares 40% amino acid identity with SPZ from Manduca sexta. Phylogenetic analysis indicated that ApSPZ might be a new member of the Spatzle type 1 family, which belongs to the Spatzle superfamily. The expression patterns of several genes involved in the Toll pathway were examined at different developmental stages and various tissues in 5th instar larvae. The examined targets included A. pernyi spatzle, GNBP, MyD88, Tolloid, cactus and dorsalA. The RT-PCR results showed that these genes were predominantly expressed in immune-responsive fat body tissue, indicating that the genes play a crucial role in A. pernyi innate immunity. Moreover, A. pernyi infection with the fungus Nosema pernyi and the gram-positive bacterium Enterococcus pernyi, but not the gram-negative bacterium Escherichia coli, activated the Toll signaling pathway. These results represent the first study of the Toll pathway in A. pernyi, which provides insight into the A. pernyi innate immune system. PMID:27483463

  7. Enhancement of the Gelation Properties of Surimi from Yellowtail Seabream (Parargyrops edita, Sparidae) with Chinese Oak Silkworm Pupa, Antheraea pernyi.

    PubMed

    Zhu, Jialin; Fan, Daming; Zhao, Jianxin; Zhang, Hao; Huang, Jianlian; Zhou, Wenguo; Zhang, Wenhai; Chen, Wei

    2016-02-01

    In this study, the textural properties and micromechanism of yellowtail seabream (Parargyrops edita, Sparidae) surimi, with and without Chinese oak silkworm pupa homogenate (SPH), were investigated at different levels. The fresh, freeze-dried, and oven-dried SPH all showed a gel-enhancing ability in suwari (40/90 °C) and modori (67/90 °C) gels, in a concentration-dependent manner. Though the drying treatments can improve the storability of SPH, compared with fresh, the effect of the active substance was weakened. Suwari and modori gels added with 5%(w/w, whole product) fresh SPH had the increase in breaking force and deformation by 37.39% and 47.98%, and 85.14% and 78.49%, respectively, compared with the control gel (without SPH addition). The major myofibrillar protein, especially myosin heavy chain (MHC), was better retained by the addition of SPH. Compared the control group, a finer, denser, and more ordered 3-dimensional gel network microstructure was obtained, and different Df (Fractal dimension) was analyzed by using the box count method. This was found in all samples from 2.838 to 2.864 for suwari gels and 2.795 to 2.857 for modori gels, respectively. Therefore, the modori of yellowtail seabream surimi, linked with endogenous proteases, could be retarded in the presence of SPH, leading to an increase in gel strength.

  8. Enhancement of the Gelation Properties of Surimi from Yellowtail Seabream (Parargyrops edita, Sparidae) with Chinese Oak Silkworm Pupa, Antheraea pernyi.

    PubMed

    Zhu, Jialin; Fan, Daming; Zhao, Jianxin; Zhang, Hao; Huang, Jianlian; Zhou, Wenguo; Zhang, Wenhai; Chen, Wei

    2016-02-01

    In this study, the textural properties and micromechanism of yellowtail seabream (Parargyrops edita, Sparidae) surimi, with and without Chinese oak silkworm pupa homogenate (SPH), were investigated at different levels. The fresh, freeze-dried, and oven-dried SPH all showed a gel-enhancing ability in suwari (40/90 °C) and modori (67/90 °C) gels, in a concentration-dependent manner. Though the drying treatments can improve the storability of SPH, compared with fresh, the effect of the active substance was weakened. Suwari and modori gels added with 5%(w/w, whole product) fresh SPH had the increase in breaking force and deformation by 37.39% and 47.98%, and 85.14% and 78.49%, respectively, compared with the control gel (without SPH addition). The major myofibrillar protein, especially myosin heavy chain (MHC), was better retained by the addition of SPH. Compared the control group, a finer, denser, and more ordered 3-dimensional gel network microstructure was obtained, and different Df (Fractal dimension) was analyzed by using the box count method. This was found in all samples from 2.838 to 2.864 for suwari gels and 2.795 to 2.857 for modori gels, respectively. Therefore, the modori of yellowtail seabream surimi, linked with endogenous proteases, could be retarded in the presence of SPH, leading to an increase in gel strength. PMID:26709730

  9. Expression and characterization of a lipase-related protein in the malpighian tubules of the Chinese oak silkworm, Antheraea pernyi.

    PubMed

    Wang, L; Li, J; Zhao, X; Qian, C; Wei, G; Zhu, B; Liu, C

    2016-10-01

    Lipases are ubiquitous enzymes in nature, which play a crucial role in fat metabolism by catalyzing the hydrolysis of triacylglycerol to free fatty acids and glycerol. However, reports concerning insect lipase are rare. In this study, we studied the expression and activity of a lipase-related protein from Antheraea pernyi (ApLRP). Recombinant ApLRP was expressed in Escherichia coli cells and used to raise rabbit anti-ApLRP polyclonal antibodies. ApLRP mRNA and protein expression were abundant in the midgut and malpighian tubules, respectively. After challenge with four different microorganisms (E. coli, Beauveria bassiana, Micrococcus luteus and nuclear polyhedrosis virus), the expression levels of ApLRP mRNA in midgut were inducted significantly compared with the control. The different pathogens induced different ApLRP gene expression patterns. The optimum temperature and pH for the enzyme's activity were 35°C and 7.0, respectively. ApLRP activity was stimulated in the presence of Mg2+, Na+, Ca2+ and b-mercaptoethanol; while Zn2+, Cu2+ and Fe3+ inhibited its activity. Detergents such as SDS, glycerol and Tween-20 increased the lipase activity by 20-30%. Our results indicated that ApLRP might play an important role in the innate immunity of insects.

  10. Expression and characterization of a lipase-related protein in the malpighian tubules of the Chinese oak silkworm, Antheraea pernyi.

    PubMed

    Wang, L; Li, J; Zhao, X; Qian, C; Wei, G; Zhu, B; Liu, C

    2016-10-01

    Lipases are ubiquitous enzymes in nature, which play a crucial role in fat metabolism by catalyzing the hydrolysis of triacylglycerol to free fatty acids and glycerol. However, reports concerning insect lipase are rare. In this study, we studied the expression and activity of a lipase-related protein from Antheraea pernyi (ApLRP). Recombinant ApLRP was expressed in Escherichia coli cells and used to raise rabbit anti-ApLRP polyclonal antibodies. ApLRP mRNA and protein expression were abundant in the midgut and malpighian tubules, respectively. After challenge with four different microorganisms (E. coli, Beauveria bassiana, Micrococcus luteus and nuclear polyhedrosis virus), the expression levels of ApLRP mRNA in midgut were inducted significantly compared with the control. The different pathogens induced different ApLRP gene expression patterns. The optimum temperature and pH for the enzyme's activity were 35°C and 7.0, respectively. ApLRP activity was stimulated in the presence of Mg2+, Na+, Ca2+ and b-mercaptoethanol; while Zn2+, Cu2+ and Fe3+ inhibited its activity. Detergents such as SDS, glycerol and Tween-20 increased the lipase activity by 20-30%. Our results indicated that ApLRP might play an important role in the innate immunity of insects. PMID:27297450

  11. Identification of immune response-related genes in the Chinese oak silkworm, Antheraea pernyi by suppression subtractive hybridization.

    PubMed

    Liu, Qiu-Ning; Zhu, Bao-Jian; Wang, Lei; Wei, Guo-Qing; Dai, Li-Shang; Lin, Kun-Zhang; Sun, Yu; Qiu, Jian-Feng; Fu, Wei-Wei; Liu, Chao-Liang

    2013-11-01

    Insects possess an innate immune system that responds to invading microorganisms. In this study, a subtractive cDNA library was constructed to screen for immune response-related genes in the fat bodies of Antheraea pernyi (Lepidoptera: Saturniidae) pupa challenged with Escherichia coli. Four hundred putative EST clones were identified by suppression subtractive hybridization (SSH), including 50 immune response-related genes, three cytoskeleton genes, eight cell cycle and apoptosis genes, five respiration and energy metabolism genes, five transport genes, 40 metabolism genes, ten stress response genes, four transcription and translation regulation genes and 77 unknown genes. To verify the reliability of the SSH data, the transcription of a set of randomly selected immune response-related genes were confirmed by semi-quantitative reverse transcription-PCR (RT-PCR) and real-time quantitative reverse transcription-PCR (qRT-PCR). These identified immune response-related genes provide insight into understanding the innate immunity in A. pernyi. PMID:24076149

  12. Mechanical properties and structure of silkworm cocoons: a comparative study of Bombyx mori, Antheraea assamensis, Antheraea pernyi and Antheraea mylitta silkworm cocoons.

    PubMed

    Zhang, J; Kaur, J; Rajkhowa, R; Li, J L; Liu, X Y; Wang, X G

    2013-08-01

    As a protective shell against environmental damage and attack by natural predators, the silkworm cocoon has outstanding mechanical properties. In particular, this multilayer non-woven composite structure can be exceptionally tough to enhance the chance of survival for silkworms while supporting their metabolic activity. Peel, out-of-plane compression and nano-indentation tests and micro-structure analysis were performed on four types of silkworm cocoon walls (domesticated Bombyx mori, semi-domesticated Antheraea assamensis and wild Antheraea pernyi and Antheraea mylitta silkworm cocoons) to understand the structure and mechanical property relationships. The wild silkworm cocoons were shown to be uniquely tough composite structures. The maximum work-of-fracture for the wild cocoons (A. pernyi and A. mylitta) was approximately 1000 J/m(2), which was almost 10 times the value for the domesticated cocoon (Bombyx mori) and 3~4 times the value for the semi-domesticated cocoon (A. assamensis). Calcium oxalate crystals were found to deposit on the outer surfaces of the semi-domesticated and wild cocoons. They did not show influence in enhancing the interlaminar adhesion between cocoon layers but exhibited much higher hardness than the cocoon pelades.

  13. Involvement of a versatile pattern recognition receptor, apolipophorin-III in prophenoloxidase activation and antibacterial defense of the Chinese oak silkworm, Antheraea pernyi.

    PubMed

    Wen, Daihua; Wang, Xialu; Shang, Lei; Huang, Yu; Li, Tienan; Wu, Chunfu; Zhang, Rong; Zhang, Jinghai

    2016-12-01

    Apolipophorin-III (apoLp-III) is an exchangeable apolipoprotein found in many insect species and functions as a lipid transport vehicle. Recent studies have shown that apoLp-III is a multifunctional molecule involved in not only lipid transportation but also innate immune responses. In the present study, the pattern recognition properties of Antheraea pernyi apoLp-III were investigated. Recombinant Ap-apoLp-III was bound to different species of microbes and further study showed the rAp-apoLp-III is capable of interacting with pathogen associated molecular patterns (PAMPs) on the microbial cell surface. In addition, an Ap-apoLp-III/PAMP mixture stimulated the prophenoloxidase (PPO) activation of A. pernyi hemolymph in vitro, to a greater extent than PAMP alone while Ap-apoLp-III itself failed to activate the PPO system, indicating that Ap-apoLp-III up-regulates PPO activation by combining with PAMP. After pathogen invasion following an injection of Staphylococcus aureus, RNAi-mediated silencing of apoLp-III decreased the transcriptional abundance of three antimicrobial peptide genes. These data suggest that apoLp-III is a versatile pattern recognition receptor and may play important roles in the innate immune responses of Antheraea pernyi. PMID:27387151

  14. Supercooling Capacity and Cold Tolerance of the Wild Silkworm, Antheraea pernyi (Lepidoptera: Saturniidae).

    PubMed

    Liu, Yan-Qun; Zheng, Xi-Xi; Ma, Hong-Fang; Xia, Run-Xi; Li, Yu-Ping; Zhang, Qi-Rui

    2016-08-01

    While wild silkworms have served humans for several thousand years, little attention on cold hardiness has been paid to these economically important species. In the present study, supercooling capacity and low temperature tolerance of Chinese oak silkworm, Antheraea pernyi (Guérin-Méneville) (Lepidoptera: Saturniidae), an economic insect reared both for silk production as well as human food, were examined under laboratory conditions. The supercooling points (SCPs) of pupae dropped significantly from a mean of -15.6°C in prediapause to -20.1°C in diapause, and then increased to -17.5°C during postdiapause development. Sex and voltinism influenced body mass but had no significant effect on the SCP. Our data demonstrated that cold tolerance of A. pernyi is tightly linked to life stage. Exposure of eggs to -5°C for up to 8 h had no effect on the hatching rate, whereas silkworm larvae failed to break through the chorion and hatch following a 4-8-h exposure to -10°C. Mean SCPs of intact eggs and naked larvae one day before hatching were similar, -23.3°C and -22.3°C, respectively, indicating that chorion does not significantly affect SCP. Comparison of lower lethal temperature (LLT50) and SCP means suggested that both pupae and eggs of A. pernyi are chill intolerant. These data will improve our understanding of low temperature tolerance in this commercially important species. PMID:27371710

  15. Supercooling Capacity and Cold Tolerance of the Wild Silkworm, Antheraea pernyi (Lepidoptera: Saturniidae).

    PubMed

    Liu, Yan-Qun; Zheng, Xi-Xi; Ma, Hong-Fang; Xia, Run-Xi; Li, Yu-Ping; Zhang, Qi-Rui

    2016-08-01

    While wild silkworms have served humans for several thousand years, little attention on cold hardiness has been paid to these economically important species. In the present study, supercooling capacity and low temperature tolerance of Chinese oak silkworm, Antheraea pernyi (Guérin-Méneville) (Lepidoptera: Saturniidae), an economic insect reared both for silk production as well as human food, were examined under laboratory conditions. The supercooling points (SCPs) of pupae dropped significantly from a mean of -15.6°C in prediapause to -20.1°C in diapause, and then increased to -17.5°C during postdiapause development. Sex and voltinism influenced body mass but had no significant effect on the SCP. Our data demonstrated that cold tolerance of A. pernyi is tightly linked to life stage. Exposure of eggs to -5°C for up to 8 h had no effect on the hatching rate, whereas silkworm larvae failed to break through the chorion and hatch following a 4-8-h exposure to -10°C. Mean SCPs of intact eggs and naked larvae one day before hatching were similar, -23.3°C and -22.3°C, respectively, indicating that chorion does not significantly affect SCP. Comparison of lower lethal temperature (LLT50) and SCP means suggested that both pupae and eggs of A. pernyi are chill intolerant. These data will improve our understanding of low temperature tolerance in this commercially important species.

  16. Cocoon of the silkworm Antheraea pernyi as an example of a thermally insulating biological interface.

    PubMed

    Jin, Xing; Zhang, Jin; Gao, Weimin; Li, Jingliang; Wang, Xungai

    2014-09-01

    Biological materials are hierarchically organized complex composites, which embrace multiple practical functionalities. As an example, the wild silkworm cocoon provides multiple protective functions against environmental and physical hazards, promoting the survival chance of moth pupae that resides inside. In the present investigation, the microstructure and thermal property of the Chinese tussah silkworm (Antheraea pernyi) cocoon in both warm and cold environments under windy conditions have been studied by experimental and numerical methods. A new computational fluid dynamics model has been developed according to the original fibrous structure of the Antheraea pernyi cocoon to simulate the unique heat transfer process through the cocoon wall. The structure of the Antheraea pernyi cocoon wall can promote the disorderness of the interior air, which increases the wind resistance by stopping most of the air flowing into the cocoon. The Antheraea pernyi cocoon is wind-proof due to the mineral crystals deposited on the outer layer surface and its hierarchical structure with low porosity and high tortuosity. The research findings have important implications to enhancing the thermal function of biomimetic protective textiles and clothing.

  17. Generation of cytotoxic molecules and oxidative stress in haemolymph of pebrinised tasar silkworm Antheraea mylitta Drury.

    PubMed

    Jena, Karmabeer; Pandey, Jay Prakash; Priya, Anshu; Kundu, Papri; Sinha, Ajit Kumar; Yadav, Harendra; Sahay, Alok

    2016-01-01

    The present study was carried out to investigate the effects of microsporidial infection on redox regulation mechanism and oxidative stress in tasar silkworm Antheraea mylitta. High level of superoxide radical (p < 0.05), nitric oxide (p < 0.001) and lipid peroxidation (p < 0.001) was observed in haemolymph of pebrinised larvae, which indicated the resultant generation of cytotoxic molecules and oxidative damage. Increased phenol oxidase (PO) activity in haemolymph of pebrinised larvae indicated the activation of immune defences during pathological conditions. In addition, higher level of glutathione-S-tranferase (GST) activity and reduced glutathione (GSH) level observed in pebrinised larvae indicated adaptive behaviour of tissue against toxic oxyradicals (p < 0.05). Conversely, low level of ascorbic acid (ASA) content suggested that the larvae might have used these compounds to counteract stress in tissues or low uptake under microspridial infection (p < 0.05). Present findings provide new insights into the cellular and biochemical bases of host-pathogen interactions in tasar silkworm A. mylitta. PMID:26930859

  18. Characterization and pathogenicity assessment of gut-associated microbes of muga silkworm Antheraea assamensis Helfer (Lepidoptera: Saturniidae).

    PubMed

    Haloi, Kishor; Kalita, Moni Kankana; Nath, Ramesh; Devi, Dipali

    2016-07-01

    Antheraea assamensis Helfer (muga silkworm) is an economically important endemic insect species of North Eastern Region of India. The silkworm is often susceptible to infection by pathogenic bacteria, leads to a disease commonly known as flacherie which causes 40% loss per annum to the silk industry. In this study, we have reported isolation, characterization and pathogenicity assessment of gut-associated bacteria of healthy and diseased muga silkworms. Thirty five bacterial isolates were screened from the gut of healthy and diseased silkworms by morphological observation and biochemical tests. 11 and 5 strains from healthy and diseased silkworm respectively were identified by 16S rRNA gene sequencing and analyzed. Pseudomonas aeruginosa (DRK1), Ornithinibacillus bavariensis (DRK2), Achromobacter xylosoxidans (KH3) and Staphylococcus aureus (FLG1) strains were commonly found in healthy as well as diseased larvae whereas, Bacillus thuringiensis (MK1) was found only in diseased larvae. Survivability analysis was performed with the identified strains by injection and oral administration (10(4)CFU/ml). The immune response of the silkworm against the pathogen was also studied by phenoloxidase and lysozyme enzyme activity assay, total and differential hemocyte count and phagocytic activity of hemocytes. It was observed that S. aureus, P. aeruginosa and B. thuringiensis significantly reduced the survivability of silkworm (p<0.001) hence found highly pathogenic. The lethal concentrations (LC50) values of the pathogenic strains were calculated at different time intervals (24, 48, 72 and 96h) within the range from 1.38×10(2) to 3.63×10(7)CFU/ml. The pathogenic groups demonstrated inhibition of phenoloxidase activity and decreased in total hemocyte count after 48h of infection. However, the lysozyme activity increased significantly in the pathogenic groups compared to the control (p<0.05). Granulocytes and plasmatocytes showed phagocytosis whereas; other types of cells did

  19. Crystallization and preliminary X-ray diffraction analysis of a protease inhibitor from the haemolymph of the Indian tasar silkworm Antheraea mylitta

    SciTech Connect

    Roy, Sobhan; Aravind, Penmatsa; Madhurantakam, Chaithanya; Ghosh, Ananta Kumar; Sankaranarayanan, Rajan; Das, Amit Kumar

    2006-07-01

    The crystallization and preliminary X-ray crystallographic analysis of a protease inhibitor from the haemolymph of the Indian tasar silk worm A. mylitta is reported. A protein with inhibitory activity against fungal proteases was purified from the haemolymph of the Indian tasar silkworm Antheraea mylitta and was crystallized using the hanging-drop vapour-diffusion method. Polyethylene glycol 3350 was used as a precipitant. Crystals belonged to space group P6{sub 3}22, with unit-cell parameters a = b = 60.6, c = 85.1 Å. X-ray diffraction data were collected and processed to a maximum resolution of 2.1 Å.

  20. Ion-induced fabrication of silk fibroin nanoparticles from Chinese oak tasar Antheraea pernyi.

    PubMed

    Wang, Juan; Zhang, Shanshan; Xing, Tieling; Kundu, Banani; Li, Mingzhong; Kundu, Subhas C; Lu, Shenzhou

    2015-08-01

    Silk protein fibroin in nanoparticles form is a promising material for drug delivery due to its pleiotropic properties, including biocompatibility, biodegradability, ease in fabrication into smaller diameters, high bioavailability, and therapeutic retention at target sites. In the present study, silk nanoparticles are fabricated from regenerated fibroin solution of the Chinese temperate oak tasar Antheraea pernyi by novel ion-induced self-assembly in a very short time under mild conditions. The resultant fibroin nanoparticles range in size from 100 to 500 nm. The molecular conformation of regenerated fibroin changes from α-helical to a β-sheet structure as a rapid function of the ionic strength and the hydrophobic and electrostatic interactions. The mild conditions are potentially advantageous for the encapsulation of sensitive drugs and therapeutic molecules such as doxorubicin hydrochloride, an amphiphilic anticancer therapeutic. In vitro release of doxorubicin from nanoparticles is pH sensitive, with approx. 65% doxorubicin remaining in the fibroin nanoparticles after 11 days. The activity of fibroin nanoparticles on hepatomas indicates the efficacy of the fibroin nanoparticles to maintain the bioactivity of the loaded doxorubicin and impart a dose-dependent cell growth inhibition. The results suggest that Chinese temperate oak tasar silk fibroin nanoparticles can be used as a sustained drug delivery vehicle.

  1. Powdered Wound Dressing Materials Made from wild Silkworm Antheraea pernyi Silk Fibroin on Full-skin Thickness Burn Wounds on Rats

    PubMed Central

    Kim, Min-Keun; Yoo, Ki-Yeon; Kwon, Kwang-Jun; Kim, Seong-Gon; Park, Young-Wook; Lee, Kwang-Gill; Jo, You-Young; Kweon, Hae-Yong

    2014-01-01

    Purpose: This study evaluated powdered burn wound dressing materials from wild silkworm fibroin in an animal model. Methods: Fifteen rats were used in this experiment. Full-thickness 2×2 cm burn wounds were created on the back of rats under anesthesia. In the two experimental groups, the wounds were treated with two different dressing materials made from silkworm fibroin. In the Control Group, natural healing without any dressing material was set as control. The wound surface area was measured at five days, seven days and 14 days. Wound healing was evaluated by histologic analysis. Results: By gross observation, there were no infections or severe inflammations through 14 days post-injury. The differences among groups were statistically significant at seven days and 14 days, postoperatively (P <0.037 and 0.001, respectively). By post hoc test, the defect size was significantly smaller in experimental Group 1 compared with the Control Group and experimental Group 2 at seven days postoperatively (P =0.022 and 0.029, respectively). The difference between Group 1 and Group 2 was statistically significant at 14 days postoperatively (P <0.001). Group 1 and control also differed significantly (P =0.002). Group 1 showed a smaller residual scar than the Control Group and Group 2 at 14 days post-injury. Histologic analysis showed more re-epithelization in Groups 1 and 2 than in the Control Groups. Conclusion: Burn wound healing was accelerated with silk fibroin spun by wild silkworm Antheraea pernyi. There was no atypical inflammation with silk dressing materials. In conclusion, silk dressing materials can be used for treatment of burn wound. PMID:27489820

  2. Genetic Characterization of a Novel Iflavirus Associated with Vomiting Disease in the Chinese Oak Silkmoth Antheraea pernyi

    PubMed Central

    Lin, Lan; de Miranda, Joachim R.; Emrich, Scott; An, Lijia; Terenius, Olle

    2014-01-01

    Larvae of the Chinese oak silkmoth (Antheraea pernyi) are often affected by AVD (A. pernyi vomiting disease), whose causative agent has long been suspected to be a virus. In an unrelated project we discovered a novel positive sense single-stranded RNA virus that could reproduce AVD symptoms upon injection into healthy A. pernyi larvae. The genome of this virus is 10,163 nucleotides long, has a natural poly-A tail, and contains a single, large open reading frame flanked at the 5′ and 3′ ends by untranslated regions containing putative structural elements for replication and translation of the virus genome. The open reading frame is predicted to encode a 3036 amino acid polyprotein with four viral structural proteins (VP1-VP4) located in the N-terminal end and the non-structural proteins, including a helicase, RNA-dependent RNA polymerase and 3C-protease, located in the C-terminal end of the polyprotein. Putative 3C-protease and autolytic cleavage sites were identified for processing the polyprotein into functional units. The genome organization, amino acid sequence and phylogenetic analyses suggest that the virus is a novel species of the genus Iflavirus, with the proposed name of Antheraea pernyi Iflavirus (ApIV). PMID:24637949

  3. Construction of a full-length cDNA Library from Chinese oak silkworm pupa and identification of a KK-42-binding protein gene in relation to pupa-diapause termination.

    PubMed

    Li, Yu-Ping; Xia, Run-Xi; Wang, Huan; Li, Xi-Sheng; Liu, Yan-Qun; Wei, Zhao-Jun; Lu, Cheng; Xiang, Zhong-Huai

    2009-06-24

    In this study we successfully constructed a full-length cDNA library from Chinese oak silkworm, Antheraea pernyi, the most well-known wild silkworm used for silk production and insect food. Total RNA was extracted from a single fresh female pupa at the diapause stage. The titer of the library was 5 x 10(5) cfu/ml and the proportion of recombinant clones was approximately 95%. Expressed sequence tag (EST) analysis was used to characterize the library. A total of 175 clustered ESTs consisting of 24 contigs and 151 singlets were generated from 250 effective sequences. Of the 175 unigenes, 97 (55.4%) were known genes but only five from A. pernyi, 37 (21.2%) were known ESTs without function annotation, and 41 (23.4%) were novel ESTs. By EST sequencing, a gene coding KK-42-binding protein in A. pernyi (named as ApKK42-BP; GenBank accession no. FJ744151) was identified and characterized. Protein sequence analysis showed that ApKK42-BP was not a membrane protein but an extracellular protein with a signal peptide at position 1-18, and contained two putative conserved domains, abhydro_lipase and abhydrolase_1, suggesting it may be a member of lipase superfamily. Expression analysis based on number of ESTs showed that ApKK42-BP was an abundant gene in the period of diapause stage, suggesting it may also be involved in pupa-diapause termination.

  4. Studies on the Influence of Host Plants and Effect of Chemical Stimulants on the Feeding Behavior in the Muga Silkworm, Antheraea assamensis

    PubMed Central

    Neog, Kartik; Unni, Balagopalan; Ahmed, Giasuddin

    2011-01-01

    The feeding habits of Antheraea assamensis, Helfer (Lepidoptera: Saturniidae) larvae towards the leaves of its four different host plants, Persea bombycina King ex. Hook (Laurales: Lauraceae), Litsea polhantha Jussieu, L. salicifolia Roxburgh ex. Nees and L. citrata Blume, and the chemical basis of feeding preference were investigated. Nutritional superiority of young and medium leaves with respect to soluble protein, total phenol and phenylalanine ammonia lyase activity was observed in the leaves of P. bombycina compared to other host plants. Attraction and feeding tests with detached leaves and artificial diet with different chemical stimulants revealed that a mixture of the flavonoids, myrcetin, and 7, 2', 4' trimethoxy dihydroxy flavone with sterol compound β-sitosterol elicited the most biting behavior by A. assamensis larvae. While linalyl acetate alone attracted larvae towards the leaves of the host plants, a mixture of caryophyllene, decyl aldehyde and dodecyl aldehyde was found to both attract them to the host leaves and cause biting behavior. Azaindole was found to deter them from the host plants. PMID:22243364

  5. Morphological and molecular characterization of Nosema pernyi, a microsporidian parasite in Antheraea pernyi.

    PubMed

    Wang, Yong; Liu, Wei; Jiang, Yiren; Huang, Ling; Irfan, Muhammad; Shi, Shenglin; Yang, Ruisheng; Qin, Li

    2015-09-01

    Nosema pernyi is a lethal pathogen that causes microsporidiosis in the Chinese oak silkworm, Antheraea pernyi. In this study, we presented its morphological and some molecular characteristics. The mature spores were measured to be 4.36 × 1.49 μm. The spore wall consisted of an electron-dense exospore (EX) and electron-lucent endospore (EN) layer. The polar filament (PF) was isofilar with 10-12 coils that were frequently arranged in a single row. Investigation results indicated that N. pernyi can infect the gut wall, silk glands, and other tissues. A full-length SMART cDNA library of N. pernyi was constructed, and then 824 expressed sequence tags (ESTs) were sequenced. Ninety unigenes, out of 197 assembled unigenes, showed significant homology to known genes of Nosema ceranae, Nosema bombycis, Encephalitozoon cuniculi, and other microsporidian species. Based on the nucleotide sequence of the α- and β-tubulin genes and amino acid sequence of actin gene, phylogenetic trees analysis showed that N. pernyi was closely related to Nosema philosamiae and Nosema antheraeae. It was correctly assigned to the Nosema group. PMID:26044882

  6. Nutrigenomics in Arma chinensis: transcriptome analysis of Arma chinensis fed on artificial diet and Chinese oak silk moth Antheraea pernyi pupae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: The insect predator, Arma chinensis (Fallou), is capable of effectively controlling many pests, such as Colorado potato beetle, cotton bollworm, and mirid bugs. Life history parameters were not as good for A. chinensis when reared on an artificial diet compared to Chinese oak silk moth p...

  7. Uptake of atmospheric carbon dioxide into silk fiber by silkworms.

    PubMed

    Magoshi, Jun; Tanaka, Toshihisa; Sasaki, Haruto; Kobayashi, Masatoshi; Magoshi, Yoshiko; Tsuda, Hidetoshi; Becker, Mary A; Inoue, Shun-ichi; Ishimaru, Ken

    2003-01-01

    The relation between the uptake of atmospheric CO(2) and insect's production of silk fiber has not yet been reported. Here, we provide the first quantitative demonstrations that four species of silkworms (Bombyx mori, Samia cynthia ricini, Antheraea pernyi, and Antheraea yamamai) and a silk-producing spider (Nephila clavata) incorporate atmospheric CO(2) into their silk fibers. The abundance of (13)C incorporated from the environment was determined by mass spectrometry and (13)C NMR measurements. Atmospheric CO(2) was incorporated into the silk fibers in the carbonyl groups of alanine, aspartic acid, serine, and glycine and the C(gamma) of aspartic acid. We show a simple model for the uptake of atmospheric CO(2) by silkworms. These results will demonstrate that silkworm has incorporated atmospheric CO(2) into silk fiber via the TCA cycle; however, the magnitude of uptake into the silk fibers is smaller than that consumed by the photosynthesis in trees and coral reefs.

  8. Nutrigenomics in Arma chinensis: Transcriptome Analysis of Arma chinensis Fed on Artificial Diet and Chinese Oak Silk Moth Antheraea pernyi Pupae

    PubMed Central

    Zou, Deyu; Coudron, Thomas A.; Liu, Chenxi; Zhang, Lisheng; Wang, Mengqing; Chen, Hongyin

    2013-01-01

    Background The insect predator, Arma chinensis, is capable of effectively controlling many pests, such as Colorado potato beetle, cotton bollworm, and mirid bugs. Our previous study demonstrated several life history parameters were diminished for A. chinensis reared on an artificial diet compared to a natural food source like the Chinese oak silk moth pupae. The molecular mechanisms underlying the nutritive impact of the artificial diet on A. chinensis health are unclear. So we utilized transcriptome information to better understand the impact of the artificial diet on A. chinensis at the molecular level. Methodology/Principal Findings Illumina HiSeq2000 was used to sequence 4.79 and 4.70 Gb of the transcriptome from pupae-fed and artificial diet-fed A. chinensis libraries, respectively, and a de novo transcriptome assembly was performed (Trinity short read assembler). This resulted in 112,029 and 98,724 contigs, clustered into 54,083 and 54,169 unigenes for pupae-fed and diet-fed A. chinensis, respectively. Unigenes from each sample’s assembly underwent sequence splicing and redundancy removal to acquire non-redundant unigenes. We obtained 55,189 unigenes of A. chinensis, including 12,046 distinct clusters and 43,143 distinct singletons. Unigene sequences were aligned by BLASTx to nr, Swiss-Prot, KEGG and COG (E-value <10−5), and further aligned by BLASTn to nt (E-value <10−5), retrieving proteins of highest sequence similarity with the given unigenes along with their protein functional annotations. Totally, 22,964, 7,898, 18,069, 15,416, 8,066 and 5,341 unigenes were annotated in nr, nt, Swiss-Prot, KEGG, COG and GO, respectively. We compared gene expression variations and found thousands of genes were differentially expressed between pupae-fed and diet-fed A. chinensis. Conclusions/Significance Our study provides abundant genomic data and offers comprehensive sequence information for studying A. chinensis. Additionally, the physiological roles of the

  9. Supplementation of an Artificial Medium for the Parasitoid Exorista larvarum (Diptera: Tachnidae) With Hemolymph of Hermetia illucens (Diptera: Stratiomyidae) or Antheraea pernyi (Lepidoptera: Saturniidae).

    PubMed

    Dindo, Maria Luisa; Vandicke, Jonas; Marchetti, Elisa; Spranghers, Thomas; Bonte, Jochem; De Clercq, Patrick

    2016-04-01

    The effect of supplementing hemolymph of the black soldier fly, Hermetia illucens (L.), or the Chinese oak silkworm, Antheraea pernyi (Guérin-Méneville), to a basic insect-free artificial medium for the tachinid Exorista larvarum (L.) was investigated. The supplementation (20% w/w) was based on the assumption that insect additives may optimize the media for this parasitoid. Egg hatch, pupal and adult yields, and sex ratio did not differ among the enriched and basic media. Preimaginal development was faster on both hemolymph-enriched media than on the basic medium. Despite the shorter development on the medium supplemented with H. illucens hemolymph than on the basic medium, on the two media puparium weights were comparable. The female flies reared on the medium enriched with H. illucens hemolymph did not lay more eggs, but the latter yielded significantly more puparia compared with the control females. Conversely, the medium enriched with A. pernyi hemolymph yielded lower female puparium weights than the basic medium and produced only one ovipositing female out of the five obtained female adults. These results indicate that the in vitro development of E. larvarum improved when the basic artificial medium was enriched with H. illucens hemolymph, whereas the supplementation with A. pernyi hemolymph negatively affected the quality of the in vitro-reared females. PMID:26921223

  10. Supplementation of an Artificial Medium for the Parasitoid Exorista larvarum (Diptera: Tachnidae) With Hemolymph of Hermetia illucens (Diptera: Stratiomyidae) or Antheraea pernyi (Lepidoptera: Saturniidae).

    PubMed

    Dindo, Maria Luisa; Vandicke, Jonas; Marchetti, Elisa; Spranghers, Thomas; Bonte, Jochem; De Clercq, Patrick

    2016-04-01

    The effect of supplementing hemolymph of the black soldier fly, Hermetia illucens (L.), or the Chinese oak silkworm, Antheraea pernyi (Guérin-Méneville), to a basic insect-free artificial medium for the tachinid Exorista larvarum (L.) was investigated. The supplementation (20% w/w) was based on the assumption that insect additives may optimize the media for this parasitoid. Egg hatch, pupal and adult yields, and sex ratio did not differ among the enriched and basic media. Preimaginal development was faster on both hemolymph-enriched media than on the basic medium. Despite the shorter development on the medium supplemented with H. illucens hemolymph than on the basic medium, on the two media puparium weights were comparable. The female flies reared on the medium enriched with H. illucens hemolymph did not lay more eggs, but the latter yielded significantly more puparia compared with the control females. Conversely, the medium enriched with A. pernyi hemolymph yielded lower female puparium weights than the basic medium and produced only one ovipositing female out of the five obtained female adults. These results indicate that the in vitro development of E. larvarum improved when the basic artificial medium was enriched with H. illucens hemolymph, whereas the supplementation with A. pernyi hemolymph negatively affected the quality of the in vitro-reared females.

  11. Interactions between fibroin and sericin proteins from Antheraea pernyi and Bombyx mori silk fibers.

    PubMed

    Du, Shan; Zhang, Jin; Zhou, Wei T; Li, Quan X; Greene, George W; Zhu, Hai J; Li, Jing L; Wang, Xun G

    2016-09-15

    Silkworm silk fibers are core-shell composites of fibroin and sericin proteins. Studying the interactions between fibroin and sericin is essential for understanding the properties of these composites. It is observed that compared to the domestic silk cocoon Bombyx mori (B. mori), the adhesion between fibroin and sericin from the wild silk cocoon, Antheraea pernyi (A. pernyi), is significantly stronger with a higher degree of heterogeneity. The adsorption of A. pernyi sericin on its fibroin is almost twice the value for B. mori sericin on fibroin, both showing a monolayer Langmuir adsorption. (1)H NMR and FTIR studies demonstrate on a molecular level the stronger interactions and the more intensive complex formation between A. pernyi fibroin and sericin, facilitated by the hydrogen bonding between glycine and serine. The findings of this study may help the design of composites with superior interfacial adhesion between different components. PMID:27314644

  12. Directional moisture transfer through a wild silkworm cocoon wall.

    PubMed

    Jin, Xing; Zhang, Jin; Gao, Weimin; Du, Shan; Li, Jingliang; Wang, Xungai

    2016-06-25

    A silkworm cocoon is a porous biological structure with multiple protective functions. In the current work, the authors have used both experimental and numerical methods to reveal the unique moisture transfer characteristics through a wild Antheraea pernyi silkworm cocoon wall, in comparison with the long-domesticated Bombyx mori silkworm cocoon walls. The water vapor transmission and water vapor permeability (WVP) properties show that the A. pernyi cocoons exhibit directional moisture transfer behavior, with easier moisture transfer from inside out than outside in [e.g., the average WVP is 0.057 g/(h m bar) from inside out and is 0.034 g/(h m bar) from outside in]. Numerical analysis shows that the cubic mineral crystals in the outer section of the A. pernyi cocoon wall create a rough surface that facilitates air turbulence and promotes disturbance amplitude of the flow field, leading to lengthened water vapor transfer path and increased tortuosity of the moist air. It also indicates the vortex of water vapor can be generated in the outer section of cocoon wall, which increases the diffusion distance of water vapor and enhances the turbulence kinetic energy and turbulence eddy dissipation, signifying higher moisture resistance in the outer section. The difference in moisture resistance of the multiple A. pernyi cocoon layers is largely responsible for the unique directional moisture transfer behavior of this wild silkworm cocoon. These findings may inspire a biomimicry approach to develop novel lightweight moisture management materials and structures.

  13. Directional moisture transfer through a wild silkworm cocoon wall.

    PubMed

    Jin, Xing; Zhang, Jin; Gao, Weimin; Du, Shan; Li, Jingliang; Wang, Xungai

    2016-01-01

    A silkworm cocoon is a porous biological structure with multiple protective functions. In the current work, the authors have used both experimental and numerical methods to reveal the unique moisture transfer characteristics through a wild Antheraea pernyi silkworm cocoon wall, in comparison with the long-domesticated Bombyx mori silkworm cocoon walls. The water vapor transmission and water vapor permeability (WVP) properties show that the A. pernyi cocoons exhibit directional moisture transfer behavior, with easier moisture transfer from inside out than outside in [e.g., the average WVP is 0.057 g/(h m bar) from inside out and is 0.034 g/(h m bar) from outside in]. Numerical analysis shows that the cubic mineral crystals in the outer section of the A. pernyi cocoon wall create a rough surface that facilitates air turbulence and promotes disturbance amplitude of the flow field, leading to lengthened water vapor transfer path and increased tortuosity of the moist air. It also indicates the vortex of water vapor can be generated in the outer section of cocoon wall, which increases the diffusion distance of water vapor and enhances the turbulence kinetic energy and turbulence eddy dissipation, signifying higher moisture resistance in the outer section. The difference in moisture resistance of the multiple A. pernyi cocoon layers is largely responsible for the unique directional moisture transfer behavior of this wild silkworm cocoon. These findings may inspire a biomimicry approach to develop novel lightweight moisture management materials and structures. PMID:27226205

  14. Molecular profiling of silkworm biodiversity in India.

    PubMed

    Chatterjee, S N; Tanushree, T

    2004-12-01

    Molecular tools opened a new vista to understand nature's bio-diversity and its relevance and the same approach was availed of to build-up the foundation work on the bio-diversity of silkworm spp in India. It is well established that the heritage of usage of silk for dress materials in India, Russia and China dates back to premedieval period and in spite of industrial development resulting to de-forestation, India still can claim as the owner of wide bio-diversity, espicially in northern India for silkworm spp. The molecular diversity was assessed among Antheraea mylitta, A. assama, A. pernyi, A. provlei, A. roylei and Philcosomia cynthia wiht 11 ISSR and 8 non-random primers on agarose gel. Nei's statistics as also Euclidean distance matrix was applied to find the genetic diversity between the six species, wherein the closest relationship between A. pernyi and A. proylei is established. With the help of POPGEN statistics, the average genetic heterozygosity appeared as 0.271 while Shanon's index is 0.4312 and alleles with segregation ratios of 3:1, 2:1, 1:1, 9:1 (generated with ISSR primers) were identified which can be utilized for future molecular breeding porgram. Further, and attempt was made to isolate a number of bands generated with 3 ISSR and six non-random primers, specific for different species and 22 such markers have been characterized through sequencing which will be made available through international public domain database.

  15. Comparison of the in vitro and in vivo degradations of silk fibroin scaffolds from mulberry and nonmulberry silkworms.

    PubMed

    You, Renchuan; Xu, Yamei; Liu, Yi; Li, Xiufang; Li, Mingzhong

    2015-02-01

    Degradation behavior is very important in the field of silk-based biomaterials. Mulberry and nonmulberry silk fibroins are structurally and functionally distinguishable; however, no studies have examined the differences in the degradation behaviors of silk materials from various silkworm species. In this study, Ca(NO3)2 was used as a uniform solvent to obtain regenerated mulberry and nonmulberry (Antheraea pernyi and Antheraea yamamai) silk fibroin (SF) solutions, and the degradation behaviors of various SF scaffolds were examined. In vitro and in vivo results demonstrated that regenerated mulberry SF scaffolds exhibited significantly higher mass loss and free amino acid content release than did nonmulberry SF scaffolds. The differences in the primary structures and condensed structures between mulberry and nonmulberry SF contributed to the significant difference in degradation rates, in which the characteristic (-Ala-)n repeats, compact crystal structure and high α-helix and β-sheet contents make nonmulberry SF more resistant than mulberry SF to enzymatic degradation. Moreover, the Antheraea pernyi and Antheraea yamamai SFs possess similar primary structures and condensed structures, although a slight difference in degradation was observed; this difference might depend on the differences in molecular weight following the regeneration process. The results indicate that the original sources of SF significantly influence the degradation rates of SF-based materials; therefore, the original sources of SF should be fully considered for preparing tissue engineering scaffolds with matched degradation rates.

  16. Genetic Diversity and Population Structure of Indian Golden Silkmoth (Antheraea assama)

    PubMed Central

    Arunkumar, Kallare P.; Sahu, Anup Kumar; Mohanty, Atish Ranjan; Awasthi, Arvind K.; Pradeep, Appukuttannair R.; Urs, S. Raje; Nagaraju, Javaregowda

    2012-01-01

    Background The Indian golden saturniid silkmoth (Antheraea assama), popularly known as muga silkmoth, is a semi-domesticated silk producing insect confined to a narrow habitat range of the northeastern region of India. Owing to the prevailing socio-political problems, the muga silkworm habitats in the northeastern region have not been accessible hampering the phylogeography studies of this rare silkmoth. Recently, we have been successful in our attempt to collect muga cocoon samples, although to a limited extent, from their natural habitats. Out of 87 microsatellite markers developed previously for A. assama, 13 informative markers were employed to genotype 97 individuals from six populations and analyzed their population structure and genetic variation. Methodology/Principal Findings We observed highly significant genetic diversity in one of the populations (WWS-1, a population derived from West Garo Hills region of Meghalaya state). Further analysis with and without WWS-1 population revealed that dramatic genetic differentiation (global FST = 0.301) was due to high genetic diversity contributed by WWS-1 population. Analysis of the remaining five populations (excluding WWS-1) showed a marked reduction in the number of alleles at all the employed loci. Structure analysis showed the presence of only two clusters: one formed by WWS-1 population and the other included the remaining five populations, inferring that there is no significant genetic diversity within and between these five populations, and suggesting that these five populations are probably derived from a single population. Patterns of recent population bottlenecks were not evident in any of the six populations studied. Conclusions/Significance A. assama inhabiting the WWS-1 region revealed very high genetic diversity, and was genetically divergent from the five populations studied. The efforts should be continued to identify and study such populations from this region as well as other muga silkworm

  17. Attachment and growth of human bone marrow derived mesenchymal stem cells on regenerated antheraea pernyi silk fibroin films.

    PubMed

    Luan, Xi-Ying; Wang, Yong; Duan, Xiang; Duan, Qiao-Yan; Li, Ming-Zhong; Lu, Shen-Zhou; Zhang, Huan-Xiang; Zhang, Xue-Guang

    2006-12-01

    Silk fibroin of the silkworm Bombyx mori has been studied extensively, while the research on Antheraea pernyi silk fibroin (A. pernyi SF) in biomaterials is only at an early stage. In this study, the attachment, morphology, growth and phenotype of human bone marrow derived mesenchymal stem cells (hBMSCs) cultured on the regenerated A. pernyi SF films were studied in vitro. The results indicated that the attachment of hBMSCs on the regenerated A. pernyi SF films was almost the same as that on the collagen films. MTT and cell counting analyses demonstrated that the growth of hBMSCs on the regenerated A. pernyi SF films was better than that on controls. Moreover, electron scanning microscopy and fluorescence-activated cell sorting assays showed that the regenerated A. pernyi SF supported hBMSCs growth and functional maintenance compared with the controls. These data suggest that the regenerated A. pernyi SF, like Bombyx mori silk fibroin (B. mori SF) and collagen, can support hBMSCs attachment, growth and phenotypic maintenance, and has better biocompatibilities for hBMSCs in vitro culture. PMID:18458403

  18. Survey and Analysis of Microsatellites in the Silkworm, Bombyx mori

    PubMed Central

    Prasad, M. Dharma; Muthulakshmi, M.; Madhu, M.; Archak, Sunil; Mita, K.; Nagaraju, J.

    2005-01-01

    We studied microsatellite frequency and distribution in 21.76-Mb random genomic sequences, 0.67-Mb BAC sequences from the Z chromosome, and 6.3-Mb EST sequences of Bombyx mori. We mined microsatellites of ≥15 bases of mononucleotide repeats and ≥5 repeat units of other classes of repeats. We estimated that microsatellites account for 0.31% of the genome of B. mori. Microsatellite tracts of A, AT, and ATT were the most abundant whereas their number drastically decreased as the length of the repeat motif increased. In general, tri- and hexanucleotide repeats were overrepresented in the transcribed sequences except TAA, GTA, and TGA, which were in excess in genomic sequences. The Z chromosome sequences contained shorter repeat types than the rest of the chromosomes in addition to a higher abundance of AT-rich repeats. Our results showed that base composition of the flanking sequence has an influence on the origin and evolution of microsatellites. Transitions/transversions were high in microsatellites of ESTs, whereas the genomic sequence had an equal number of substitutions and indels. The average heterozygosity value for 23 polymorphic microsatellite loci surveyed in 13 diverse silkmoth strains having 2–14 alleles was 0.54. Only 36 (18.2%) of 198 microsatellite loci were polymorphic between the two divergent silkworm populations and 10 (5%) loci revealed null alleles. The microsatellite map generated using these polymorphic markers resulted in 8 linkage groups. B. mori microsatellite loci were the most conserved in its immediate ancestor, B. mandarina, followed by the wild saturniid silkmoth, Antheraea assama. PMID:15371363

  19. Investigation into the Effect of Altitude on the Differential Hemocyte Count of Circulating Plasmatocytes and Granulocytes of Larval Stage of Antheraea assama (Lepidoptera: Saturniidae).

    PubMed

    Baishya, Bhavna Prishnee; Bardoloi, Sunayan; Bharali, Rupjyoti

    2015-01-01

    Differential hemocyte count of circulating plasmatocytes (PL) and granulocytes (GR) of fifth-instar larvae of Muga Silkworm Antheraea assama Westwood (Lepidoptera: Saturniidae) reared at four different sericulture farms situated at different altitudes, viz, Khanapara State Sericulture Farm, Assam, altitude 55.5 m above sea level (ASL); Nongpoh (Central Silk Board farm), Meghalaya, altitude 464 m ASL; Tura (Central Silk Board farm), West Garo Hills, Meghalaya, 657 m ASL; and Kalimpong (Central Silk Board farm), West Bengal, altitude 1,247 m ASL, were calculated and compared to investigate the effect of altitude on the number of PL and GR per mm(3) of larval hemolymph. The investigation showed that the mean circulating PL and GR were highest at Khanapara (55.5 m ASL) located at the lowest altitude, whereas their numbers gradually decreased with increase in altitude at Nongpoh (464 m ASL), Tura (657 m ASL), and Kalimpong (1,247 m ASL). This may be attributed to the average environmental temperatures observed at different altitudes, which may affect the overall hemocyte load of larval stages reared at those altitudes.

  20. Sericin Composition in the Silk of Antheraea yamamai.

    PubMed

    Zurovec, Michal; Yonemura, Naoyuki; Kludkiewicz, Barbara; Sehnal, František; Kodrik, Dalibor; Vieira, Ligia Cota; Kucerova, Lucie; Strnad, Hynek; Konik, Peter; Sehadova, Hana

    2016-05-01

    The silks produced by caterpillars consist of fibroin proteins that form two core filaments, and sericin proteins that seal filaments into a fiber and conglutinate fibers in the cocoon. Sericin genes are well-known in Bombyx mori (Bombycidae) but have received little attention in other insects. This paper shows that Antheraea yamamai (Saturniidae) contains five sericin genes very different from the three sericin genes of B. mori. In spite of differences, all known sericins are characterized by short exons 1 and 2 (out of 3-12 exons), expression in the middle silk gland section, presence of repeats with high contents of Ser and charged amino acid residues, and secretion as a sticky silk component soluble in hot water. The B. mori sericins represent tentative phylogenetic lineages (I) BmSer1 and orthologs in Saturniidae, (II) BmSer2, and (III) BmSer3 and related sericins of Saturniidae and of the pyralid Galleria mellonella. The lineage (IV) seems to be limited to Saturniidae. Concerted evolution of the sericin genes was apparently associated with gene amplifications as well as gene loses. Differences in the silk fiber morphology indicate that the cocktail of sericins linking the filaments and coating the fiber is modified during spinning. Silks are composite biomaterials of conserved function in spite of great diversity of their composition. PMID:27049111

  1. Genetic analysis of Indian tasar silkmoth (Antheraea mylitta) populations.

    PubMed

    Chakraborty, Saikat; Muthulakshmi, M; Vardhini, Deena; Jayaprakash, P; Nagaraju, J; Arunkumar, K P

    2015-01-01

    Indian tasar silkmoth, Antheraea mylitta is an economically important wild silkmoth species distributed across India. A number of morphologically and ethologically well-defined ecotypes are known for this species that differ in their primary food plant specificity. Most of these ecotypes do not interbreed in nature, but are able to produce offspring under captive conditions. Microsatellite markers were developed for A. mylitta, and out of these, ten well-behaved microsatellite loci were used to analyze the population structure of different ecoraces. A total of 154 individual moths belonging to eight different ecoraces, were screened at each locus. Hierarchical analysis of population structure using Analysis of MOlecular VAriance (AMOVA) revealed significant structuring (FST = 0.154) and considerable inbreeding (FIS = 0.505). A significant isolation by distance was also observed. The number of possible population clusters was investigated using distance method, Bayesian algorithm and self organization maps (SOM). The first two methods revealed two distinct clusters, whereas the SOM showed the different ecoraces not to be clearly differentiated. These results suggest that although there is a large degree of phenotypic variation among the different ecoraces of A. mylitta, genetically they are not very different, and the phenotypic differences may largely be a result of their respective ecology. PMID:26510465

  2. Genetic analysis of Indian tasar silkmoth (Antheraea mylitta) populations

    PubMed Central

    Chakraborty, Saikat; Muthulakshmi, M; Vardhini, Deena; Jayaprakash, P; Nagaraju, J; Arunkumar, K. P.

    2015-01-01

    Indian tasar silkmoth, Antheraea mylitta is an economically important wild silkmoth species distributed across India. A number of morphologically and ethologically well-defined ecotypes are known for this species that differ in their primary food plant specificity. Most of these ecotypes do not interbreed in nature, but are able to produce offspring under captive conditions. Microsatellite markers were developed for A. mylitta, and out of these, ten well-behaved microsatellite loci were used to analyze the population structure of different ecoraces. A total of 154 individual moths belonging to eight different ecoraces, were screened at each locus. Hierarchical analysis of population structure using Analysis of MOlecular VAriance (AMOVA) revealed significant structuring (FST = 0.154) and considerable inbreeding (FIS = 0.505). A significant isolation by distance was also observed. The number of possible population clusters was investigated using distance method, Bayesian algorithm and self organization maps (SOM). The first two methods revealed two distinct clusters, whereas the SOM showed the different ecoraces not to be clearly differentiated. These results suggest that although there is a large degree of phenotypic variation among the different ecoraces of A. mylitta, genetically they are not very different, and the phenotypic differences may largely be a result of their respective ecology. PMID:26510465

  3. Strain Rate and Anisotropic Microstructure Dependent Mechanical Behaviors of Silkworm Cocoon Shells

    PubMed Central

    Xu, Jun; Zhang, Wen; Gao, Xiang; Meng, Wanlin; Guan, Juan

    2016-01-01

    Silkworm cocoons are multi-layered composite structures comprised of high strength silk fiber and sericin, and their mechanical properties have been naturally selected to protect pupas during metamorphosis from various types of external attacks. The present study attempts to gain a comprehensive understanding of the mechanical properties of cocoon shell materials from wild silkworm species Antheraea pernyi under dynamic loading rates. Five dynamic strain rates from 0.00625 s-1 to 12.5 s-1 are tested to show the strain rate sensitivity of the cocoon shell material. In the meantime, the anisotropy of the cocoon shell is considered and the cocoon shell specimens are cut along 0°, 45° and 90° orientation to the short axis of cocoons. Typical mechanical properties including Young’s modulus, yield strength, ultimate strength and ultimate strain are extracted and analyzed from the stress-strain curves. Furthermore, the fracture morphologies of the cocoon shell specimens are observed under scanning electron microscopy to help understand the relationship between the mechanical properties and the microstructures of the cocoon material. A discussion on the dynamic strain rate effect on the mechanical properties of cocoon shell material is followed by fitting our experimental results to two previous models, and the effect could be well explained. We also compare natural and dried cocoon materials for the dynamic strain rate effect and interestingly the dried cocoon shells show better overall mechanical properties. This study provides a different perspective on the mechanical properties of cocoon material as a composite material, and provides some insight for bio-inspired engineering materials. PMID:26939063

  4. Strain Rate and Anisotropic Microstructure Dependent Mechanical Behaviors of Silkworm Cocoon Shells.

    PubMed

    Xu, Jun; Zhang, Wen; Gao, Xiang; Meng, Wanlin; Guan, Juan

    2016-01-01

    Silkworm cocoons are multi-layered composite structures comprised of high strength silk fiber and sericin, and their mechanical properties have been naturally selected to protect pupas during metamorphosis from various types of external attacks. The present study attempts to gain a comprehensive understanding of the mechanical properties of cocoon shell materials from wild silkworm species Antheraea pernyi under dynamic loading rates. Five dynamic strain rates from 0.00625 s-1 to 12.5 s-1 are tested to show the strain rate sensitivity of the cocoon shell material. In the meantime, the anisotropy of the cocoon shell is considered and the cocoon shell specimens are cut along 0°, 45° and 90° orientation to the short axis of cocoons. Typical mechanical properties including Young's modulus, yield strength, ultimate strength and ultimate strain are extracted and analyzed from the stress-strain curves. Furthermore, the fracture morphologies of the cocoon shell specimens are observed under scanning electron microscopy to help understand the relationship between the mechanical properties and the microstructures of the cocoon material. A discussion on the dynamic strain rate effect on the mechanical properties of cocoon shell material is followed by fitting our experimental results to two previous models, and the effect could be well explained. We also compare natural and dried cocoon materials for the dynamic strain rate effect and interestingly the dried cocoon shells show better overall mechanical properties. This study provides a different perspective on the mechanical properties of cocoon material as a composite material, and provides some insight for bio-inspired engineering materials. PMID:26939063

  5. Spectral imaging analysis for silkworm gender classification

    NASA Astrophysics Data System (ADS)

    Sumriddetchkajorn, Sarun; Kamtongdee, Chakkrit; Sa-Ngiamsak, Chiranut

    2013-05-01

    We examine the effect of different wavelength spectra in the performance of our optical penetration-based silkworm pupa sex identification system. With available low-cost light emitting diodes (LEDs), each emitting different wavelength spectra at 468 nm, 565 nm, 639 nm, 940 nm, and broad white light, we find that the body of the silkworm pupa can block blue and near infrared light while allowing green and red light pass through. In particular, the red light can clearly highlight an important organ called "chitin gland" of the female, leading to high accuracy of silkworm gender identification. In our experiment with 120 silkworm pupae, measured high average 92.8% and lower average 87.5% accuracies in identifying silkworm gender are obtained under red and white light LEDs, respectively.

  6. [EPR spectra of silkworm eggs].

    PubMed

    Korkhova, E D; Chepel', L M; Nikolov, O T; Komar', I N; Shakhbazov, V G

    1976-01-01

    ESR spectra of the native grain of the silkworm have been studied in the course of embryo formation, during a diapause, and during embryo development after the diapause. It is shown that the nature of ESR spectra of the grain is not determined by the metabolic processes, but by the presence of pigments in it and other stationary biological structures having developed pi-systems and unpaired electrones. The latter are mainly found in the envelope and may give the ESR spectra, with various g-factors. A dependence of the ESR spectra integral intensity of the grain on the denotype is discovered.

  7. Silkworm protein: its possibility as an actuator

    NASA Astrophysics Data System (ADS)

    Jin, Hyoung-Joon; Myung, Seung Jun; Kim, Heung Soo; Jung, Woochul; Kim, Jaehwan

    2006-03-01

    The possibility of silkworm (Bombyx mori) protein as a base material of biomimetic actuator was investigated in this paper. Silkworm films were prepared from high concentrations of regenerated fibroin in aqueous solution. Films with thickness of about 100 μm were prepared for coating electrodes. The cast silk films were coated by very thin gold electrode on both sides of the film. Tensile test of cast film showed bi-modal trend, which is typical stress-strain relation of polymeric film. As the test of a possible biomimetic actuator, silkworm film actuator provides bending deformations according to the magnitude and frequency of the applied electric filed. Although the present bending deformation of silkworm film actuator is smaller than that of Electro-Active Paper actuator, it provides the possibility of biomimetic actuator.

  8. Biomimetic nucleation of hydroxyapatite crystals mediated by Antheraea pernyi silk sericin promotes osteogenic differentiation of human bone marrow derived mesenchymal stem cells.

    PubMed

    Yang, Mingying; Shuai, Yajun; Zhang, Can; Chen, Yuyin; Zhu, Liangjun; Mao, Chuanbin; OuYang, Hongwei

    2014-04-14

    Biomacromolecules have been used as templates to grow hydroxyapatite crystals (HAps) by biomineralization to fabricate mineralized materials for potential application in bone tissue engineering. Silk sericin is a protein with features desirable as a biomaterial, such as increased hydrophilicity and biodegradation. Mineralization of the silk sericin from Antheraea pernyi (A. pernyi) silkworm has rarely been reported. Here, for the first time, nucleation of HAps on A. pernyi silk sericin (AS) was attempted through a wet precipitation method and consequently the cell viability and osteogenic differentiation of BMSCs on mineralized AS were investigated. It was found that AS mediated the nucleation of HAps in the form of nanoneedles while self-assembling into β-sheet conformation, leading to the formation of a biomineralized protein based biomaterial. The cell viability assay of BMSCs showed that the mineralization of AS stimulated cell adhesion and proliferation, showing that the resultant AS biomaterial is biocompatible. The differentiation assay confirmed that the mineralized AS significantly promoted the osteogenic differentiation of BMSCs when compared to nonmineralized AS as well as other types of sericin (B. mori sericin), suggesting that the resultant mineralized AS biomaterial has potential in promoting bone formation. This result represented the first work proving the osteogenic differentiation of BMSCs directed by silk sericin. Therefore, the biomineralization of A. pernyi silk sericin coupled with seeding BMSCs on the resultant mineralized biomaterials is a useful strategy to develop the potential application of this unexplored silk sericin in the field of bone tissue engineering. This study lays the foundation for the use of A. pernyi silk sericin as a potential scaffold for tissue engineering.

  9. Biomimetic Nucleation of Hydroxyapatite Crystals Mediated by Antheraea pernyi Silk Sericin Promotes Osteogenic Differentiation of Human Bone Marrow Derived Mesenchymal Stem Cells

    PubMed Central

    2015-01-01

    Biomacromolecules have been used as templates to grow hydroxyapatite crystals (HAps) by biomineralization to fabricate mineralized materials for potential application in bone tissue engineering. Silk sericin is a protein with features desirable as a biomaterial, such as increased hydrophilicity and biodegradation. Mineralization of the silk sericin from Antheraea pernyi (A. pernyi) silkworm has rarely been reported. Here, for the first time, nucleation of HAps on A. pernyi silk sericin (AS) was attempted through a wet precipitation method and consequently the cell viability and osteogenic differentiation of BMSCs on mineralized AS were investigated. It was found that AS mediated the nucleation of HAps in the form of nanoneedles while self-assembling into β-sheet conformation, leading to the formation of a biomineralized protein based biomaterial. The cell viability assay of BMSCs showed that the mineralization of AS stimulated cell adhesion and proliferation, showing that the resultant AS biomaterial is biocompatible. The differentiation assay confirmed that the mineralized AS significantly promoted the osteogenic differentiation of BMSCs when compared to nonmineralized AS as well as other types of sericin (B. mori sericin), suggesting that the resultant mineralized AS biomaterial has potential in promoting bone formation. This result represented the first work proving the osteogenic differentiation of BMSCs directed by silk sericin. Therefore, the biomineralization of A. pernyi silk sericin coupled with seeding BMSCs on the resultant mineralized biomaterials is a useful strategy to develop the potential application of this unexplored silk sericin in the field of bone tissue engineering. This study lays the foundation for the use of A. pernyi silk sericin as a potential scaffold for tissue engineering. PMID:24666022

  10. Female Attacus atlas respond to pheromones of Antheraea polyphemus: a comparative electrophysiological and biochemical study.

    PubMed

    Maida, R; Ziesmann, J

    2001-01-01

    Female Attacus atlas respond electrophysiologically to both of the Antheraea polyphemus pheromone components (E,Z)-6,11-hexadecadienyl acetate and (E,Z)-6,11-hexadecadienal. Moreover, they possess a pheromone-binding protein (PBP) and general odorant-binding proteins (GOBPs), as well as a pheromone-degrading sensillar esterase and aldehyde oxidase enzymes. They show no electroantennogram responses to their own gland extract. In contrast, female A. polyphemus do not respond to their own or to A. atlas pheromone. Male A. atlas do not detect any of the A. polyphemus compounds but only the conspecific female gland extracts. Both male A. atlas and female A. polyphemus possess PBP and GOBP but lack the pheromone-degrading esterases of male Antheraea. The results indicate that the two species use quite distinct classes of chemicals as pheromones. In spite of this, the N-terminal amino acid sequences of the PBPs show homology of 68%.

  11. Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The development of a spider silk manufacturing process is of great interest. piggyBac vectors were used to create transgenic silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk prote...

  12. Comparative transcriptomic analysis of silkwormBmovo-1 and wild type silkworm ovary

    PubMed Central

    Xue, Renyu; Hu, Xiaolong; Zhu, Liyuan; Cao, Guangli; Huang, Moli; Xue, Gaoxu; Song, Zuowei; Lu, Jiayu; Chen, Xueying; Gong, Chengliang

    2015-01-01

    The detailed molecular mechanism of Bmovo-1 regulation of ovary size is unclear. To uncover the mechanism of Bmovo-1 regulation of ovarian development and oogenesis using RNA-Seq, we compared the transcriptomes of wild type (WT) and Bmovo-1-overexpressing silkworm (silkworm+Bmovo-1) ovaries. Using a pair-end Illumina Solexa sequencing strategy, 5,296,942 total reads were obtained from silkworm+Bmovo-1 ovaries and 6,306,078 from WT ovaries. The average read length was about 100 bp. Clean read ratios were 98.79% for silkworm+Bmovo-1 and 98.87% for WT silkworm ovaries. Comparative transcriptome analysis showed 123 upregulated and 111 downregulated genes in silkworm+Bmovo-1 ovaries. These differentially expressed genes were enriched in the extracellular and extracellular spaces and involved in metabolism, genetic information processing, environmental information processing, cellular processes and organismal systems. Bmovo-1 overexpression in silkworm ovaries might promote anabolism for ovarian development and oogenesis and oocyte proliferation and transport of nutrients to ovaries by altering nutrient partitioning, which would support ovary development. Excessive consumption of nutrients for ovary development alters nutrient partitioning and deters silk protein synthesis. PMID:26643037

  13. Detection and characterization of Wolbachia infection in silkworm

    PubMed Central

    Zha, Xingfu; Zhang, Wenji; Zhou, Chunyan; Zhang, Liying; Xiang, Zhonghuai; Xia, Qingyou

    2014-01-01

    Wolbachia naturally infects a wide variety of arthropods, where it plays important roles in host reproduction. It was previously reported that Wolbachia did not infect silkworm. By means of PCR and sequencing we found in this study that Wolbachia is indeed present in silkworm. Phylogenetic analysis indicates that Wolbachia infection in silkworm may have occurred via transfer from parasitic wasps. Furthermore, Southern blotting results suggest a lateral transfer of the wsp gene into the genomes of some wild silkworms. By antibiotic treatments, we found that tetracycline and ciprofloxacin can eliminate Wolbachia in the silkworm and Wolbachia is important to ovary development of silkworm. These results provide clues towards a more comprehensive understanding of the interaction between Wolbachia and silkworm and possibly other lepidopteran insects. PMID:25249781

  14. Combination solar hothouse and silkworm breeding house

    SciTech Connect

    Vardiashvili, A.B.; Muradov, M.; Kim, V.D.

    1980-01-01

    The basic arrangement is shown for a combination silkworm breeding house and solar hothouse with subsoil irrigation and accumulation of heat; it employs a semicylindrical film covering. The process of accumulation of solar heat in the subsoil pebble stores, in water-heater banks, and in the soil is described.

  15. Silk Spinning in Silkworms and Spiders

    PubMed Central

    Andersson, Marlene; Johansson, Jan; Rising, Anna

    2016-01-01

    Spiders and silkworms spin silks that outcompete the toughness of all natural and manmade fibers. Herein, we compare and contrast the spinning of silk in silkworms and spiders, with the aim of identifying features that are important for fiber formation. Although spiders and silkworms are very distantly related, some features of spinning silk seem to be universal. Both spiders and silkworms produce large silk proteins that are highly repetitive and extremely soluble at high pH, likely due to the globular terminal domains that flank an intermediate repetitive region. The silk proteins are produced and stored at a very high concentration in glands, and then transported along a narrowing tube in which they change conformation in response primarily to a pH gradient generated by carbonic anhydrase and proton pumps, as well as to ions and shear forces. The silk proteins thereby convert from random coil and alpha helical soluble conformations to beta sheet fibers. We suggest that factors that need to be optimized for successful production of artificial silk proteins capable of forming tough fibers include protein solubility, pH sensitivity, and preservation of natively folded proteins throughout the purification and initial spinning processes. PMID:27517908

  16. Silk Spinning in Silkworms and Spiders.

    PubMed

    Andersson, Marlene; Johansson, Jan; Rising, Anna

    2016-01-01

    Spiders and silkworms spin silks that outcompete the toughness of all natural and manmade fibers. Herein, we compare and contrast the spinning of silk in silkworms and spiders, with the aim of identifying features that are important for fiber formation. Although spiders and silkworms are very distantly related, some features of spinning silk seem to be universal. Both spiders and silkworms produce large silk proteins that are highly repetitive and extremely soluble at high pH, likely due to the globular terminal domains that flank an intermediate repetitive region. The silk proteins are produced and stored at a very high concentration in glands, and then transported along a narrowing tube in which they change conformation in response primarily to a pH gradient generated by carbonic anhydrase and proton pumps, as well as to ions and shear forces. The silk proteins thereby convert from random coil and alpha helical soluble conformations to beta sheet fibers. We suggest that factors that need to be optimized for successful production of artificial silk proteins capable of forming tough fibers include protein solubility, pH sensitivity, and preservation of natively folded proteins throughout the purification and initial spinning processes. PMID:27517908

  17. Silk Spinning in Silkworms and Spiders.

    PubMed

    Andersson, Marlene; Johansson, Jan; Rising, Anna

    2016-08-09

    Spiders and silkworms spin silks that outcompete the toughness of all natural and manmade fibers. Herein, we compare and contrast the spinning of silk in silkworms and spiders, with the aim of identifying features that are important for fiber formation. Although spiders and silkworms are very distantly related, some features of spinning silk seem to be universal. Both spiders and silkworms produce large silk proteins that are highly repetitive and extremely soluble at high pH, likely due to the globular terminal domains that flank an intermediate repetitive region. The silk proteins are produced and stored at a very high concentration in glands, and then transported along a narrowing tube in which they change conformation in response primarily to a pH gradient generated by carbonic anhydrase and proton pumps, as well as to ions and shear forces. The silk proteins thereby convert from random coil and alpha helical soluble conformations to beta sheet fibers. We suggest that factors that need to be optimized for successful production of artificial silk proteins capable of forming tough fibers include protein solubility, pH sensitivity, and preservation of natively folded proteins throughout the purification and initial spinning processes.

  18. Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties

    PubMed Central

    Teulé, Florence; Miao, Yun-Gen; Sohn, Bong-Hee; Kim, Young-Soo; Hull, J. Joe; Fraser, Malcolm J.; Lewis, Randolph V.; Jarvis, Donald L.

    2012-01-01

    The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk proteins integrated in an extremely stable manner. Furthermore, these composite fibers were, on average, tougher than the parental silkworm silk fibers and as tough as native dragline spider silk fibers. These results demonstrate that silkworms can be engineered to manufacture composite silk fibers containing stably integrated spider silk protein sequences, which significantly improve the overall mechanical properties of the parental silkworm silk fibers. PMID:22215590

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

    PubMed

    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. PMID:22239062

  20. Fruit body formation on silkworm by Cordyceps militaris

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Injection inoculation protocols for fruit body formation of Cordyceps militaris were investigated to improve the incidence of infection in the silkworm species Bombyx mori. Injection, with suspensions of C. militaris hyphal bodies into living silkworm pupae, was used to test for fruit body productio...

  1. Calcium Oxalate Accumulation in Malpighian Tubules of Silkworm (Bombyx mori)

    NASA Astrophysics Data System (ADS)

    Wyman, Aaron J.; Webb, Mary Alice

    2007-04-01

    Silkworm provides an ideal model system for study of calcium oxalate crystallization in kidney-like organs, called Malpighian tubules. During their growth and development, silkworm larvae accumulate massive amounts of calcium oxalate crystals in their Malpighian tubules with no apparent harm to the organism. This manuscript reports studies of crystal structure in the tubules along with analyses identifying molecular constituents of tubule exudate.

  2. Rearing of silkworm under hypobaric and hypoxia conditions

    NASA Astrophysics Data System (ADS)

    Hashimoto, Hirofumi; Nakayama, Shin; Yamashita, Masamichi; Space Agriculture Task Force, J.

    In order to investigate of a possibility of utilizing silkworm for the space agriculture, rearing of silkworms was examined under hypobaric and hypoxia conditions. In terms of structural mechanics, the lower inner pressure of Martian greenhouse has advantage to reduce requirements on physical properties of mechanical member of the pressurized structure. The main objective of this study is to know the influence of lower total pressure and hypoxia condition on silkworm. Silkworms are reared under following four hypobaric and hypoxia conditions, 10kPa pure oxygen, 20kPa pure oxygen, 10kPa oxygen and 10kPa nitrogen, and 10kPa oxygen and 90kPa nitrogen. After rearing them to pupa stage, growth of silkworms was found poor under all hypobaric hypoxia conditions compared to those grown under the normal atmospheric condition; the control group. The growth under total pressure of 20kPa is slightly fast.

  3. Silkworm expression system as a platform technology in life science.

    PubMed

    Kato, Tatsuya; Kajikawa, Mizuho; Maenaka, Katsumi; Park, Enoch Y

    2010-01-01

    Many recombinant proteins have been successfully produced in silkworm larvae or pupae and used for academic and industrial purposes. Several recombinant proteins produced by silkworms have already been commercialized. However, construction of a recombinant baculovirus containing a gene of interest requires tedious and troublesome steps and takes a long time (3-6 months). The recent development of a bacmid, Escherichia coli and Bombyx mori shuttle vector, has eliminated the conventional tedious procedures required to identify and isolate recombinant viruses. Several technical improvements, including a cysteine protease or chitinase deletion bacmid and chaperone-assisted expression and coexpression, have led to significantly increased protein yields and reduced costs for large-scale production. Terminal N-acetyl glucosamine and galactose residues were found in the N-glycan structures produced by silkworms, which are different from those generated by insect cells. Genomic elucidation of silkworm has opened a new chapter in utilization of silkworm. Transgenic silkworm technology provides a stable production of recombinant protein. Baculovirus surface display expression is one of the low-cost approaches toward silkworm larvae-derived recombinant subunit vaccines. The expression of pharmaceutically relevant proteins, including cell/viral surface proteins, membrane proteins, and guanine nucleotide-binding protein (G protein) coupled receptors, using silkworm larvae or cocoons has become very attractive. Silkworm biotechnology is an innovative and easy approach to achieve high protein expression levels and is a very promising platform technology in the field of life science. Like the "Silkroad," we expect that the "Bioroad" from Asia to Europe will be established by the silkworm expression system. PMID:19830419

  4. Mechanical behaviour and formation process of silkworm silk gut.

    PubMed

    Cenis, José L; Madurga, Rodrigo; Aznar-Cervantes, Salvador D; Lozano-Pérez, A Abel; Marí-Buyé, Núria; Meseguer-Olmo, Luis; Plaza, Gustavo R; Guinea, Gustavo V; Elices, Manuel; Del Pozo, Francisco; Pérez-Rigueiro, José

    2015-12-14

    High performance silk fibers were produced directly from the silk glands of silkworms (Bombyx mori) following an alternative route to natural spinning. This route is based on a traditional procedure that consists of soaking the silk glands in a vinegar solution and stretching them by hand leading to the so called silkworm guts. Here we present, to the authors' best knowledge, the first comprehensive study on the formation, properties and microstructure of silkworm gut fibers. Comparison of the tensile properties and microstructural organization of the silkworm guts with those of naturally spun fibers allows gain of a deeper insight into the mechanisms that lead to the formation of the fiber, as well as the relationship between the microstructure and properties of these materials. In this regard, it is proved that an acidic environment and subsequent application of tensile stress in the range of 1000 kPa are sufficient conditions for the formation of a silk fiber.

  5. Genome segment 5 of Antheraea mylitta cytoplasmic polyhedrosis virus encodes a bona fide guanylyltransferase

    PubMed Central

    2014-01-01

    Background Antheraea mylitta cytoplasmic polyhedrosis virus (AmCPV), a cypovirus of Reoviridae family, infects non mulberry Indian silk worm, Antheraea mylitta, and contains eleven segmented double stranded RNA in its genome (S1-S11). Some of its genome segments (S1-S3, and S6-S11) have been previously characterized but genome segment encoding the viral guanylyltransferase which helps in RNA capping has not been characterized. Results In this study genome segment 5 (S5) of AmCPV was converted to cDNA, cloned and sequenced. S5 consisted of 2180 nucleotides, with one long ORF of 1818 nucleotides and could encode a protein of 606 amino acids with molecular mass of ~65 kDa (p65). Bioinformatics analysis showed presence of KLRS and HxnH motifs as observed in some other reoviral guanylyltransferase and suggests that S5 may encodes viral guanylyltransferase. The ORF of S5 was expressed in E. coli as 65 kDa his tagged fusion protein, purified through Ni-NTA chromatography and polyclonal antibody was raised. Immunoblot analysis of virion particles with the purified antibody showed specific immunoreactive band and suggests p65 as a viral structural protein. Functional analysis showed that recombinant p65 possesses guanylyltransferase activity, and transfers GMP moiety to the 5' diphosphate (A/G) ended viral RNA after the formation of p65-GMP complex for capping. Kinetic analysis showed Km of this enzyme for GTP and RNA was 34.24 uM and 98.35 nM, respectively. Site directed mutagenesis at K21A in KLRS motif, and H93A or H105A in HxnH motif completely abolished the autoguanylylation activity and indicates importance of these residues at these sites. Thermodynamic analysis showed p65-GTP interaction was primarily driven by enthalpy (ΔH = -399.1 ± 4.1 kJ/mol) whereas the p65-RNA interaction by favorable entropy (0.043 ± 0.0049 kJ/ mol). Conclusion Viral capping enzymes play a critical role in the post transcriptional or post replication modification in case of

  6. A comparison of virus genome sequences with their host silkworm, Bombyx mori.

    PubMed

    Tang, Xu-Dong; Yue, Ya-Jie; Wang, Wei; Li, Nan; Shen, Zhong-Yuan

    2016-01-15

    With the recent availability of the genomes of many viruses and the silkworm, Bombyx mori, as well as a variety of Basic Local Alignment Search Tool (BLAST) programs, a new opportunity to gain insight into the interaction of viruses with the silkworm is possible. This study aims to determine the possible existence of sequence identities between the genomes of viruses and the silkworm and attempts to explain this phenomenon. BLAST searches of the genomes of viruses against the silkworm genome were performed using the resources of the National Center for Biotechnology Information. All studied viruses contained variable numbers of short regions with sequence identity to the genome of the silkworm. The short regions of sequence identity in the genome of the silkworm may be derived from the genomes of viruses in the long history of silkworm-virus interaction. This study is the first to compare these genomes, and may contribute to research on the interaction between viruses and the silkworm.

  7. Usefulness of silkworm as a host animal for understanding pathogenicity of Cryptococcus neoformans.

    PubMed

    Ishii, Masaki; Matsumoto, Yasuhiko; Sekimizu, Kazuhisa

    2016-02-01

    We propose Cryptococcus neoformans infection model using silkworm for understanding cryptococcosis and screening of therapeutically effective antibiotics. Silkworm is an insect whose rearing methods were established through a long history of the sericulture industry. Silkworm facilitates experiments using a large number of individuals because of low cost for rearing and few ethical problems caused by killing animals. Silkworm can be reared at 37˚C to perform infection experiments at same temperature to human body. Injection of accurate amounts of samples into hemolymph of silkworm by usual syringes is easy to be done since silkworm has an appropriate size to handle. Moreover two injection methods, injection into hemolymph and intestine, are distinguishable for silkworms. The former is correspondent to intravenous injection, and the latter is to oral administration in humans. Taking these advantages of silkworms as host animals, it is possible to evaluate the virulence factors in C. neoformans and the therapeutic efficacy of antifungal agents. PMID:26902902

  8. Effects of fluoride in mulberry leaves on the growth and development of silkworm

    SciTech Connect

    Wang Chia-hsi; Qian Da-fu; Li Zheng-fang; Gao Xu-ping

    1980-01-01

    The effects of fluorides on mulberry and silkworm were investigated. The results had shown that polluted mulberry leaves which contain more than 30 parts per million fluorides (dry wt.) may induce acute damage to silkworm. 6 tables.

  9. Genome segment 6 of Antheraea mylitta cypovirus encodes a structural protein with ATPase activity

    SciTech Connect

    Chavali, Venkata R.M.; Madhurantakam, Chaithanya; Ghorai, Suvankar; Roy, Sobhan; Das, Amit K.; Ghosh, Ananta K.

    2008-07-20

    The genome segment 6 (S6) of the 11 double stranded RNA genomes from Antheraea mylitta cypovirus was converted into cDNA, cloned and sequenced. S6 consisted of 1944 nucleotides with an ORF of 607 amino acids and could encode a protein of 68 kDa, termed P68. Motif scan and molecular docking analysis of P68 showed the presence of two cystathionine beta synthase (CBS) domains and ATP binding sites. The ORF of AmCPV S6 was expressed in E. coli as His-tag fusion protein and polyclonal antibody was raised. Immunoblot analysis of virus infected gut cells and purified polyhedra using raised anti-p68 polyclonal antibody showed that S6 encodes a viral structural protein. Fluorescence and ATPase assay of soluble P68 produced in Sf-9 cells via baculovirus expression system showed its ability to bind and cleave ATP. These results suggest that P68 may bind viral RNA through CBS domains and help in replication and transcription through ATP binding and hydrolysis.

  10. Antheraea pernyi silk fibroin for targeted gene delivery of VEGF165-Ang-1 with PEI.

    PubMed

    Ma, Caili; Lv, Linlin; Liu, Yu; Yu, Yanni; You, Renchuan; Yang, Jicheng; Li, Mingzhong

    2014-06-01

    Vascularization is a crucial challenge in tissue engineering. One solution for this problem is to implant scaffolds that contain functional genes that promote vascularization by providing angiogenic growth factors via a gene delivery carrier. Poly(ethylenimine) (PEI) is a gene delivery carrier with high transfection efficiency but with cytotoxicity. To solve this problem, we utilized Antheraea pernyi silk fibroin (ASF), which has favorable cytocompatibility and biodegradability, RGD sequences and a negative charge, in conjunction with PEI, as the delivery vector for vascular endothelial growth factor (VEGF) 165-angiopoietin-1 (Ang-1) dual gene simultaneous expression plasmid, creating an ASF/PEI/pDNA complex. The results suggested that the zeta potential of the ASF/PEI/pDNA complex was significantly lower than that of the PEI/pDNA complex. Decreased nitrogen and increased oxygen on the surface of the complex demonstrated that the ASF had successfully combined with the surface of the PEI/pDNA. Furthermore, the complexes resisted digestion by nucleic acid enzymes and degradation by serum. L929 cells were cultured and transfected in vitro and improved cytotoxicity was found when the cells were transfected with ASF/PEI/pDNA compared with PEI/pDNA. In addition, the transfection efficiency and VEGF secretion increased. In general, this study provides a novel method for decreasing the cytotoxicity of PEI gene delivery vectors and increasing transfection efficiency of angiogenesis-related genes.

  11. Structure and properties of regenerated Antheraea pernyi silk fibroin in aqueous solution.

    PubMed

    Tao, Wei; Li, Mingzhong; Zhao, Chunxia

    2007-04-10

    Antheraea pernyi silk fibroin fibers were dissolved by aqueous lithium thiocyanate to obtain regenerated A. pernyi silk fibroin solution. By means of circular dichroism, (13)C NMR and Raman spectroscopy, the molecular conformation of regenerated A. pernyi silk fibroin in aqueous solution was investigated. The relationship of environmental factors and sol-gel transformation behavior of regenerated A. pernyi silk fibroin was also studied. The molecular conformations of regenerated A. pernyi silk fibroin mainly were alpha-helix and random coil in solution. There also existed a little beta-sheet conformation. It was obviously different with Bombyx mori silk fibroin, whose molecular conformation in solution was only random coil but no alpha-helix existence. With the increase of temperature and solution concentration and with the decrease of solution pH value, the gelation velocity of regenerated A. pernyi silk fibroin solution increased. Especially, it showed that A. pernyi silk fibroin was more sensitive to temperature than B. mori silk fibroin during the sol-gel transformation. The velocity increased obviously when the temperature was above 30 degrees C. During the sol-gel transformation, the molecular conformation of regenerated A. pernyi silk fibroin changed from random coil to beta-sheet structure. The results of these studies provided important insight into the preparation of new biomaterials by silk fibroin protein.

  12. Molecular architecture of silk fibroin of Indian golden silkmoth, Antheraea assama

    PubMed Central

    Gupta K, Adarsh; Mita, Kazuei; Arunkumar, Kallare P.; Nagaraju, Javaregowda

    2015-01-01

    The golden silk spun by Indian golden silkmoth Antheraea assama, is regarded for its shimmering golden luster, tenacity and value as biomaterial. This report describes the gene coding for golden silk H-fibroin (AaFhc), its expression, full-length sequence and structurally important motifs discerning the underlying genetic and biochemical factors responsible for its much sought-after properties. The coding region, with biased isocodons, encodes highly repetitious crystalline core, flanked by a pair of 5′ and 3′ non-repetitious ends. AaFhc mRNA expression is strictly territorial, confined to the posterior silk gland, encoding a protein of size 230 kDa, which makes homodimers making the elementary structural units of the fibrous core of the golden silk. Characteristic polyalanine repeats that make tight β-sheet crystals alternate with non-polyalanine repeats that make less orderly antiparallel β-sheets, β-turns and partial α-helices. Phylogenetic analysis of the conserved N-terminal amorphous motif and the comparative analysis of the crystalline region with other saturniid H-fibroins reveal that AaFhc has longer, numerous and relatively uniform repeat motifs with lower serine content that assume tighter β-crystals and denser packing, which are speculated to be responsible for its acclaimed properties of higher tensile strength and higher refractive index responsible for golden luster. PMID:26235912

  13. UV-reflecting wing scales in the silk moth Antheraea assamensis: its biophysical implications.

    PubMed

    Dey, Sudip; Singh, Surendra; Dey, Samrat; Chaudhury, Sudipto; Chakraborty, Rahul; Hooroo, R N K; Sharma, D K

    2011-01-01

    Scanning electron microcopy of the wing scales from some specific locations in the silk moth, Antheraea assamensis, revealed the presence of 50-60-nm-thick microridges and 80-nm-thick air spaces between them. The principle of optics suggests that when the measured average thickness of microridges or lamellae is about 50-60 nm, that of the air-space between them is about 80-82 nm, then due to the known refractive index of air (1.0) and the infrared refractive index of lamellar cuticle (1.60), the lamellae (microridges) and the intervening lamellar air spaces will have similar optical thickness. A common optical thickness for the two types of layers causes the microridge structures to function in the fashion of a quarter wavelength interference filter and to reflect UV light. Behavioral studies under the natural conditions and short experimental exposure of the moths to artificial UV light suggests that coupling behavior of the moth is governed to a significant extent by these UV-reflecting wing scales. The importance of the study in overcoming a major problem of low autocoupling efficiency of moths in the Muga Silk Industry is discussed with the help of relevant literature.

  14. Structure and physical properties of silkworm cocoons

    PubMed Central

    Chen, Fujia; Porter, David; Vollrath, Fritz

    2012-01-01

    Silkworm cocoons have evolved a wide range of different structures and combinations of physical and chemical properties in order to cope with different threats and environmental conditions. We present our observations and measurements on 25 diverse types of cocoons in a first attempt to correlate physical properties with the structure and morphology of the cocoons. These two architectural parameters appear to be far more important than the material properties of the silk fibres themselves. We consider tensile and compressive mechanical properties and gas permeation of the cocoon walls, and in each case identify mechanisms or models that relate these properties to cocoon structure, usually based upon non-woven fibre composites. These properties are of relevance also for synthetic non-woven composite materials and our studies will help formulate bio-inspired design principles for new materials. PMID:22552916

  15. Current use of silkworm larvae (Bombyx mori) as an animal model in pharmaco-medical research.

    PubMed

    Nwibo, Don Daniel; Hamamoto, Hiroshi; Matsumoto, Yasuhiko; Kaito, Chikara; Sekimizu, Kazuhisa

    2015-04-01

    We advocate the use of silkworm larvae, Bombyx mori, as an animal model for discovery of drug candidates. We have established several disease models using silkworms, which offer technical advantages in drug development and the study of host-pathogen interaction. This mini review briefly describes recent trends in the use of silkworm larvae as a non-mammalian model for drug discovery and it offers suggestions regarding the potential for silkworm use in pharmaceutical-biomedical research.

  16. A hyperglycemic silkworm model for evaluating hypoglycemic activity of Rehmanniae Radix, an herbal medicine.

    PubMed

    Matsumoto, Yasuhiko; Sekimizu, Kazuhisa

    2016-02-01

    Silkworm shows hyperglycemia after intake of diet containing large amount of glucose. The hyperglycemic silkworm model is useful for evaluation of anti-diabetic drugs. A hot water extract of Rehmanniae Radix, an herbal medicine, showed hypoglycemic effect against the hyperglycemic silkworms. This method is applicable for quick and simple evaluation of the hypoglycemic activities of different batches of Rehmanniae Radix. Our findings suggest that silkworms have a lot of merit as experimental animals for evaluation of various herbal medicines. PMID:26902904

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

    PubMed

    Zabelina, Valeriya; Klymenko, Vyacheslav; Tamura, Toshiki; Doroshenko, Karina; Liang, Haoyuan; Sezutsu, Hideki; Sehnal, František

    2015-09-01

    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 for commercial use. 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 as genetically stable female stocks whose unfertilized eggs are induced to develop by heat-shock treatment. Any exclusively female generation contains exact copies of the maternal clone-founder genome. Ovaries transplanted in either direction between the standard and the parthenogenetic genotypes yield eggs capable of parthenocloning. In addition, use ofmale larvae as ovary recipients eliminates diapause in eggs produced in the implants. Unfertilized eggs of some silkworm clones respond also to the cold-shock treatment by producing homozygous fertile sons; cloned females can be crossed with their parthenogenetic sons to obtain progeny homozygous for the transgene in both sexes. Rational exploitation of available parthenozygous pools and the use of parthenocloning methods enable rapid fixation and maintenance of the desired genotypes.

  18. Accumulation of 1-deoxynojirimycin in silkworm, Bombyx mori L.

    PubMed Central

    Yin, Hao; Shi, Xin-qin; Sun, Bo; Ye, Jing-jing; Duan, Zu-an; Zhou, Xiao-ling; Cui, Wei-zheng; Wu, Xiao-feng

    2010-01-01

    1-deoxynojirimycin (1-DNJ) contents in the silkworm, Bombyx mori, at different developmental stages and tissues were investigated by using reverse-phase high-performance liquid chromatography. The 1-DNJ contents of silkworm larvae change significantly with their developmental stages. The male larvae showed higher accumulation efficiency of 1-DNJ than the females and also a significant variation was observed among the silkworm strains. The present results show that tissue distribution of 1-DNJ was significantly higher in blood, digestive juice, and alimentary canal, but no 1-DNJ was observed in the silkgland. Moreover, 1-DNJ was not found in silkworms fed with artificial diet that does not contain mulberry leaf powder. This proves that silkworms obtain 1-DNJ from mulberry leaves; they could not synthesize 1-DNJ by themselves. The accumulation and excretion of 1-DNJ change periodically during the larval stage. There was no 1-DNJ in the newly-hatched larvae and 1-DNJ was mainly accumulated during the early and middle stages of every instar, while excreted at later stages of larval development. Further, it is possible to extract 1-DNJ from the larval feces and it is optimal to develop the 1-DNJ related products for diabetic auxiliary therapy. PMID:20349525

  19. Engineering Silkworms for Resistance to Baculovirus Through Multigene RNA Interference

    PubMed Central

    Subbaiah, Edupalli V.; Royer, Corinne; Kanginakudru, Sriramana; Satyavathi, Valluri V.; Babu, Adari Sobhan; Sivaprasad, Vankadara; Chavancy, Gérard; DaRocha, Martine; Jalabert, Audrey; Mauchamp, Bernard; Basha, Ibrahim; Couble, Pierre; Nagaraju, Javaregowda

    2013-01-01

    Bombyx mori nucleopolyhedrovirus (BmNPV) that infects the silkworm, B. mori, accounts for >50% of silk cocoon crop losses globally. We speculated that simultaneous targeting of several BmNPV essential genes in transgenic silkworm would elicit a stable defense against the virus. We introduced into the silkworm germline the vectors carrying short sequences of four essential BmNPV genes in tandem, either in sense or antisense or in inverted-repeat arrangement. The transgenic silkworms carrying the inverted repeat-containing transgene showed stable protection against high doses of baculovirus infection. Further, the antiviral trait was incorporated to a commercially productive silkworm strain highly susceptible to BmNPV. This led to combining the high-yielding cocoon and silk traits of the parental commercial strain and a very high level of refractoriness (>75% survival rate as compared to <15% in nontransgenic lines) to baculovirus infection conferred by the transgene. We also observed impaired infectivity of the occlusion bodies derived from the transgenic lines as compared to the wild-type ones. Currently, large-scale exploitation of these transgenic lines is underway to bring about economic transformation of sericulture. PMID:23105011

  20. In vitro study of the proteolytic degradation of Antheraea pernyi silk fibroin.

    PubMed

    Taddei, Paola; Arai, Takayuki; Boschi, Alessandra; Monti, Patrizia; Tsukada, Masuhiro; Freddi, Giuliano

    2006-01-01

    In this study, Antheraea pernyi silk fibroin (Ap-SF) films were incubated with Protease Type XXI from Streptomyces griseus, at 37 degrees C, to investigate the degradation behavior in an in vitro model system. The enzyme-resistant fractions of Ap-SF films and the soluble peptides formed by proteolytic degradation were collected at specified times, from 1 to 17 days, and analyzed by high performance liquid chromatography, differential scanning calorimetry, FT-Raman, and FT-IR spectroscopy. Proteolysis resulted in extensive weight loss and progressive fragmentation of films, especially at long degradation times. A range of soluble peptides was formed by proteolysis. By high performance-size exclusion chromatography it was found that their average molecular weight changed with the time of incubation. The chemical analysis of the enzyme-resistant fraction of Ap-SF films at different times of degradation indicated that the proteolytic attack preferentially occurred in the less ordered Gly rich sequences and that the contribution of the Ala rich crystalline regions to the composition of biodegraded films became progressively larger. Accordingly, DSC and spectroscopic results showed an enhancement of the crystalline character of the biodegraded films. From the behavior of the most important thermal transitions, it was deduced that the alpha-helix domains probably represent the most enzyme-resistant fraction. The in vitro approach used in the present study seems to be a valid tool for studying the rate and mechanism of degradation of Ap-SF films and of other biopolymers of potential biomedical utility.

  1. Spermine-modified Antheraea pernyi silk fibroin as a gene delivery carrier

    PubMed Central

    Yu, Yanni; Hu, Yongpei; Li, Xiufang; Liu, Yu; Li, Mingzhong; Yang, Jicheng; Sheng, Weihua

    2016-01-01

    The development of a novel cationized polymer used as a gene delivery carrier that can conveniently and effectively transfect cells resulting in a stably expressed target gene remains a challenge. Antheraea pernyi silk fibroin (ASF) is a cytocompatible and biodegradable natural polymer, and it possesses Arg–Gly–Asp sequences but a negative charge. In order to render ASF amenable to packaging plasmid DNA (pDNA), spermine was used to modify ASF to synthesize cationized ASF (CASF), which was used as a gene delivery carrier. CASF was characterized using trinitrobenzene sulfonic acid assay, the zeta potential determination, and a Fourier transform infrared analysis, and the results of these characterizations indicated that the –NH2 in spermine effectively reacts with the –COOH in the side chains of ASF. Spermine grafted to the side chains of ASF resulted in the conversion of the negative charge of ASF to a positive charge. CASF packaged pDNA and formed CASF/pDNA complexes, which exhibited spherical morphology with average particle sizes of 215–281 nm and zeta potential of approximately +3.0 mV to +3.2 mV. The results of the MTT assay, confocal laser scanning microscopy, and flow cytometry analysis in a human endothelial cell line revealed that CASF/pDNA complexes exhibited lower cytotoxicity and higher transfection efficiency compared to the pDNA complexes of polyethyleneimine. These results indicate that our synthesized CASF, a cationized polymer, is a potential gene delivery carrier with the advantages of biodegradability and low cytotoxicity. PMID:27042056

  2. Genome segment 4 of Antheraea mylitta cytoplasmic polyhedrosis virus encodes RNA triphosphatase and methyltransferases.

    PubMed

    Biswas, Poulomi; Kundu, Anirban; Ghosh, Ananta Kumar

    2015-01-01

    Cloning and sequencing of Antheraea mylitta cytoplasmic polyhedrosis virus (AmCPV) genome segment S4 showed that it consists of 3410 nt with a single ORF of 1110 aa which could encode a protein of ~127 kDa (p127). Bioinformatics analysis showed the presence of a 5' RNA triphosphatase (RTPase) domain (LRDR), a S-adenosyl-l-methionine (SAM)-binding (GxGxG) motif and the KDKE tetrad of 2'-O-methyltransferase (MTase), which suggested that S4 may encode RTPase and MTase. The ORF of S4 was expressed in Escherichia coli as a His-tagged fusion protein and purified by nickel-nitrilotriacetic acid affinity chromatography. Biochemical analysis of recombinant p127 showed its RTPase as well as SAM-dependent guanine N(7)-and ribose 2'-O-MTase activities. A MTase assay using in vitro transcribed AmCPV S2 RNA having a 5' G*pppG end showed that guanine N(7) methylation occurred prior to the ribose 2'-O methylation to yield a m(7)GpppG/m(7)GpppGm RNA cap. Mutagenesis of the SAM-binding (GxGxG) motif (G831A) completely abolished N(7)- and 2'-O-MTase activities, indicating the importance of these residues for capping. From the kinetic analysis, the Km values of N(7)-MTase for SAM and RNA were calculated as 4.41 and 0.39 µM, respectively. These results suggested that AmCPV S4-encoded p127 catalyses RTPase and two cap methylation reactions for capping the 5' end of viral RNA. PMID:25228490

  3. Identification and evolution of the orphan genes in the domestic silkworm, Bombyx mori.

    PubMed

    Sun, Wei; Zhao, Xin-Wei; Zhang, Ze

    2015-09-14

    Orphan genes (OGs) which have no recognizable homology to any sequences in other species could contribute to the species specific adaptations. In this study, we identified 738 OGs in the silkworm genome. About 31% of the silkworm OGs is derived from transposable elements, and 5.1% of the silkworm OGs emerged from gene duplication followed by divergence of paralogs. Five de novo silkworm OGs originated from non-coding regions. Microarray data suggested that most of the silkworm OGs were expressed in limited tissues. RNA interference experiments suggested that five de novo OGs are not essential to the silkworm, implying that they may contribute to genetic redundancy or species-specific adaptation. Our results provide some new insights into the evolutionary significance of the silkworm OGs.

  4. Suppression of intestinal immunity through silencing of TCTP by RNAi in transgenic silkworm, Bombyx mori.

    PubMed

    Hu, Cuimei; Wang, Fei; Ma, Sanyuan; Li, Xianyang; Song, Liang; Hua, Xiaoting; Xia, Qingyou

    2015-12-10

    Intestinal immune response is a front line of host defense. The host factors that participate in intestinal immunity response remain largely unknown. We recently reported that Translationally Controlled Tumor Protein (BmTCTP) was obtained by constructing a phage display cDNA library of the silkworm midgut and carrying out high throughput screening of pathogen binding molecules. To further address the function of BmTCTP in silkworm intestinal immunity, transgenic RNAi silkworms were constructed by microinjection piggBac plasmid to Dazao embryos. The antimicrobial capacity of transgenic silkworm decreased since the expression of gut antimicrobial peptide from transgenic silkworm was not sufficiently induced during oral microbial challenge. Moreover, dynamic ERK phosphorylation from transgenic silkworm midgut was disrupted. Taken together, the innate immunity of intestinal was suppressed through disruption of dynamic ERK phosphorylation after oral microbial infection as a result of RNAi-mediated knockdown of midgut TCTP in transgenic silkworm.

  5. Phylogenetics of eggshell morphogenesis in Antheraea (lepidoptera: saturniidae): unique origin and repeated reduction of the aeropyle crown.

    PubMed

    Regier, Jerome C; Paukstadt, Ulrich; Paukstadt, Laela H; Mitter, Charles; Peigler, Richard S

    2005-04-01

    Integrated phylogenetic and developmental analyses should enhance our understanding of morphological evolution and thereby improve systematists' ability to utilize morphological characters, but case studies are few. The eggshell (chorion) of Lepidoptera (Insecta) has proven especially tractable experimentally for such analyses because its morphogenesis proceeds by extracellular assembly of proteins. This study focuses on a morphological novelty, the aeropyle crown, that arises at the end of choriogenesis in the wild silkmoth genus Antheraea. Aeropyle crowns are cylindrical projections, ending in prominent prongs, that surround the openings of breathing tubes (aeropyle channels) traversing the chorion. They occur over the entire egg surface in some species, are localized to a circumferential band in many others, and in some are missing entirely, thus exhibiting variation typical of discrete characters analyzed in morphological phylogenetics. Seeking an integrated developmental-phylogenetic view, we first survey aeropyle crown variation broadly across Antheraea and related genera. We then map these observations onto a robust phylogeny, based on three nuclear genes, to test the adequacy of character codings for aeropyle crown variation and to estimate the frequency and direction of change in those characters. Thirdly, we draw on previous studies of choriogenesis, supplemented by new data on gene expression, to hypothesize developmental-genetic bases for the inferred chorion character transformations. Aeropyle crowns are inferred to arise just once, in the ancestor of Antheraea, but to undergo four or more subsequent reductions without regain, a pattern consistent with Dollo's Law. Spatial distribution shows an analogous trend, though less clear-cut, toward reduction of coverage by aeropyle crowns. These trends suggest either that there is little or no natural selection on the details of the aeropyle crown structure or that evolution toward functional optima is ongoing

  6. From silkworms to bees: Diseases of beneficial insects

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The diseases of the silkworm (Bombyx mori) and managed bees, including the honey bee (Apis mellifera), bumbles bees (Bombus spp.), the alfalfa leafcutting bee (Megachile rotundata), and mason bees (Osmia spp.) are reviewed, with diagnostic descriptions and a summary of control methods for production...

  7. Feeding scenario of the silkworm Bombyx Mori, L. in the BLSS

    NASA Astrophysics Data System (ADS)

    Yu, XiaoHui; Liu, Hong; Tong, Ling

    A simple subunit of the bioregenerative life support system (BLSS) consisting of the ground-controlled mulberry ( Morus alba L.) and the silkworms was set up on the ground. The mulberry tree could provide nutrient mulberry fruits for astronauts and its leaves as the main feedstuff for the silkworms until their third instar. Astronauts utilized curled lettuce ( Lactuca sativa L.) stem as vegetables and the silkworms over third instar could be fed on 65% of inedible leaves of the lettuce. About 71.4% of protein were detected in the silkworm larval powder; thus, 105 silkworms could satisfy the requirement of one person per day. Besides, 18 kinds of amino acids were determined in the obtained silkworm powder. Moreover, the R-criterion was suggested to estimate and optimize the animal feeding facilities. The scenario of treating the wastes is also proposed in this paper. Our results may be valuable for the establishment of a complex BLSS in the future.

  8. Effects of different Bombyx mori silkworm varieties on the structural characteristics and properties of silk.

    PubMed

    Chung, Da Eun; Kim, Hyung Hwan; Kim, Moo Kon; Lee, Ki Hoon; Park, Young Hwan; Um, In Chul

    2015-08-01

    Silk has attracted the attention of biomedical researchers because of its good biocompatibility. Although various characteristics of silk are needed for its successful application in biomedical fields, the performance of silk material is limited. Although there are many varieties of Bombyx mori silkworm, the effect of different silkworm varieties on regenerated silk has not been considered in detail. That is, the use of a diverse variety of silkworms has not been considered in non-textile applications resulting in limited performance of silk materials. In this study, the effects of different silkworm varieties on the structural characteristics and properties of silk cocoon and regenerated silk fibroin (SF) were examined. Structural characteristics of silk cocoon including color, fiber diameter, and porosity, differed depending on the silkworm variety. Furthermore, molecular weight, solution viscosity, and mechanical properties of regenerated SF were influenced by the variety of silkworm, while the amino acid composition, β-sheet crystallization by formic acid, and cyto-compatibility of regenerated SF did not differ between the samples from different varieties of silkworm. These results imply that diverse performance of silk can be obtained by controlling the silkworm variety, and that the use of different varieties of silkworm might be a good way to strengthen the performance of silk in biomedical fields.

  9. Effects of different Bombyx mori silkworm varieties on the structural characteristics and properties of silk.

    PubMed

    Chung, Da Eun; Kim, Hyung Hwan; Kim, Moo Kon; Lee, Ki Hoon; Park, Young Hwan; Um, In Chul

    2015-08-01

    Silk has attracted the attention of biomedical researchers because of its good biocompatibility. Although various characteristics of silk are needed for its successful application in biomedical fields, the performance of silk material is limited. Although there are many varieties of Bombyx mori silkworm, the effect of different silkworm varieties on regenerated silk has not been considered in detail. That is, the use of a diverse variety of silkworms has not been considered in non-textile applications resulting in limited performance of silk materials. In this study, the effects of different silkworm varieties on the structural characteristics and properties of silk cocoon and regenerated silk fibroin (SF) were examined. Structural characteristics of silk cocoon including color, fiber diameter, and porosity, differed depending on the silkworm variety. Furthermore, molecular weight, solution viscosity, and mechanical properties of regenerated SF were influenced by the variety of silkworm, while the amino acid composition, β-sheet crystallization by formic acid, and cyto-compatibility of regenerated SF did not differ between the samples from different varieties of silkworm. These results imply that diverse performance of silk can be obtained by controlling the silkworm variety, and that the use of different varieties of silkworm might be a good way to strengthen the performance of silk in biomedical fields. PMID:26072984

  10. Initial ground experiments of silkworm cultures living on different feedstock for provision of high quality animal protein for human in space

    NASA Astrophysics Data System (ADS)

    Yang, Yunan; Tang, Liman; Tong, Ling; Liu, Yang; Liu, Hong; Li, Xiaomin

    2010-09-01

    Silkworm could be an alternative to provide edible animal protein in Controlled Ecological Life Support System (CELSS) for long-term manned space missions. Silkworms can consume non-edible plant residue and convert plant nutrients to high quality edible animal protein for astronauts. The preliminary investigation of silkworm culture was carried out in earth environment. The silkworms were fed with artificial silkworm diet and the leaves of stem lettuce ( Lactuca sativa L. var. angustana Irish) separately and the nutritional structure of silkworm was investigated and compared, The culture experiments showed that: (1) Stem lettuce leaves could be used as food of silkworm. The protein content of silkworm fed with lettuce leaves can reach 70% of dry mass. (2) The protein content of silkworm powder produced by the fifth instar silkworm larvae was 70%, which was similar to the protein content of silkworm pupae. The powder of the fifth instar silkworm larvae can be utilized by astronaut. (3) The biotransformation rate of silkworm larvae between the third instar and the fifth instar could reach above 70%. The biotransformation cycle of silkworm was determined as 24 days. (4) Using the stem lettuce leaves to raise silkworm, the coarse fiber content of silkworm excrements reached about 33%. The requirements of space silkworm culture equipment, feeding approaches and feeding conditions were also preliminarily designed and calculated. It is estimated that 2.2 m 3 of culture space could satisfy daily animal protein demand for seven astronauts.

  11. Nutrient composition and respiration characteristics of silkworms in the Bioregenerative Life Support System

    NASA Astrophysics Data System (ADS)

    Tong, Ling; Yu, Xiaohui; Liu, Hong

    As the appropriate space animal candidate, silkworm(Bombyx Mori L.) can supply animal food for taikonauts and consume inedible parts of plants in Bioregenerative Life Support Sys-tem(BLSS). Due to the features of BLSS, the silkworm breeding method in the system differ-ent from the conventional one is feeding the silkworm in the first three developing stages with mulberry leaves and with lettuce leaves in the latter two developing stages. Therefore, it is nec-essary to investigate the biochemical components and respiration characteristics of silkworms raised with this method to supply data bases for the inclusion of silkworms in the system to conduct system experiments. The nutrient compositions of silkworm powder (SP) which are the grinded and freeze-dried silkworm on the 3rd day in the fifth developing stage containing protein, fat, vitamins, minerals and fatty acids were determined with international standard analyzing methods in this study. The results showed that SP was rich in protein and amino acids. There were twelve kinds of essential vitamins, nine kinds of minerals and twelve kinds of fatty acids in SP. In contrast, SP had much better nutrient components than snail, fish, chicken, beef and pork as animal food for crew members. Moreover, 359 kCal can be generated per 100g of SP (dry weight). The respirations of silkworm during its whole growing process under two main physiological statuses which were eating and non-eating leaves were studied. According to the results measured by the animal respiration measuring system, there were much difference among the respirations of silkworms under the two main physiological statuses. The amounts of O2 inhaled and CO2 exhaled by the silkworms when they were eating leaves were more than those under the non-eating status. Even under the same status, the respiration characteristics of silkworms in five different developing stages were also different from one an-other. The respiratory quotients of silkworms under two

  12. Aroma potential of oak battens prepared from decommissioned oak barrels.

    PubMed

    Li, Sijing; Crump, Anna M; Grbin, Paul R; Cozzolino, Daniel; Warren, Peter; Hayasaka, Yoji; Wilkinson, Kerry L

    2015-04-01

    During barrel maturation, volatile compounds are extracted from oak wood and impart aroma and flavor to wine, enhancing its character and complexity. However, barrels contain a finite pool of extractable material, which diminishes with time. As a consequence, most barrels are decommissioned after 5 or 6 years. This study investigated whether or not decommissioned barrels can be "reclaimed" and utilized as a previously untapped source of quality oak for wine maturation. Oak battens were prepared from staves of decommissioned French and American oak barrels, and their composition analyzed before and after toasting. The oak lactone glycoconjugate content of untoasted reclaimed oak was determined by liquid chromatography-tandem mass spectrometry, while the concentrations of cis- and trans-oak lactone, guaiacol, 4-methlyguaiacol, vanillin, eugenol, furfural, and 5-methylfurfural present in toasted reclaimed oak were determined by gas chromatography-mass spectrometry. Aroma potential was then evaluated by comparing the composition of reclaimed oak with that of new oak. Comparable levels of oak lactone glycoconjugates and oak volatiles were observed, demonstrating the aroma potential of reclaimed oak and therefore its suitability as a raw material for alternative oak products, i.e., chips or battens, for the maturation of wine. The temperature profiles achieved during toasting were also measured to evaluate the viability of any yeast or bacteria present in reclaimed oak. PMID:25771908

  13. Establishment of the BacMam system using silkworm baculovirus.

    PubMed

    Imai, Atsutoshi; Tadokoro, Takashi; Kita, Shunsuke; Horiuchi, Masataka; Fukuhara, Hideo; Maenaka, Katsumi

    2016-09-16

    The BacMam system uses modified insect viruses (baculoviruses) as vehicles to efficiently deliver genes for expression in mammalian cells. The technique can be widely applied to large-scale recombinant protein production with appropriate modifications, high-throughput screening platforms for cell-based assays, and the delivery of large genes. The silkworm system is often employed as a rapid and cost-effective approach for recombinant baculovirus generation. Here we have developed the novel BacMam system using silkworm baculovirus, and shown the successful expression of EGFP in mammalian cells. The transduction to mammalian cells via the BacMam system was improved by adding phosphate-buffered saline and sodium butyrate to the culture medium and lowering the temperature after viral infection. This study provides an alternative gene delivery system for mammalian cells, which has various potential applications, including efficient native protein production and gene therapy. PMID:27480929

  14. Transcriptome Analysis of Thermal Parthenogenesis of the Domesticated Silkworm

    PubMed Central

    Du, Xin; Yao, Lusong; Li, Fengbo; Meng, Zhiqi

    2015-01-01

    Thermal induction of parthenogenesis (also known as thermal parthenogenesis) in silkworms is an important technique that has been used in artificial insemination, expansion of hybridization, transgenesis and sericultural production; however, the exact mechanisms of this induction remain unclear. This study aimed to investigate the gene expression profile in silkworms undergoing thermal parthenogenesis using RNA-seq analysis. The transcriptome profiles indicated that in non-induced and induced eggs, the numbers of differentially expressed genes (DEGs) for the parthenogenetic line (PL) and amphigenetic line (AL) were 538 and 545, respectively, as determined by fold-change ≥ 2. Gene ontology (GO) analysis showed that DEGs between two lines were mainly involved in reproduction, formation of chorion, female gamete generation and cell development pathways. Upregulation of many chorion genes in AL suggests that the maturation rate of AL eggs was slower than PL eggs. Some DEGs related to reactive oxygen species removal, DNA repair and heat shock response were differentially expressed between the two lines, such as MPV-17, REV1 and HSP68. These results supported the view that a large fraction of genes are differentially expressed between PL and AL, which offers a new approach to identifying the molecular mechanism of silkworm thermal parthenogenesis. PMID:26274803

  15. Optical penetration-based silkworm pupa gender sensor structure.

    PubMed

    Sumriddetchkajorn, Sarun; Kamtongdee, Chakkrit

    2012-02-01

    This paper proposes and experimentally demonstrates for what is believed to be the first time a highly sought-after optical structure for highly-accurate identification of the silkworm pupa gender. The key idea is to exploit a long wavelength optical beam in the red or near infrared spectrum that can effectively and safely penetrate the body of a silkworm pupa. Later on, simple image processing operations via image thresholding, blob filtering, and image inversion processes are applied in order to eliminate the unwanted image noises and at the same time highlight the gender gland. Experimental proof of concept using three 636 nm wavelength light emitting diodes, a two-dimensional web camera, an 8 bit microcontroller board, and a notebook computer shows a very high 95.6% total accuracy in identifying the gender of 45 silkworm pupae with a measured fast identification time of 96.6 ms. Other key features include low cost, low component counts, and ease of implementation and control.

  16. Animal welfare and use of silkworm as a model animal.

    PubMed

    Sekimizu, N; Paudel, A; Hamamoto, H

    2012-08-01

    Sacrificing model animals is required for developing effective drugs before being used in human beings. In Japan today, at least 4,210,000 mice and other mammals are sacrificed to a total of 6,140,000 per year for the purpose of medical studies. All the animals treated in Japan, including test animals, are managed under control of "Act on Welfare and Management of Animals". Under the principle of this Act, no person shall kill, injure, or inflict cruelty on animals without due cause. "Animal" addressed in the Act can be defined as a "vertebrate animal". If we can make use of invertebrate animals in testing instead of vertebrate ones, that would be a remarkable solution for the issue of animal welfare. Furthermore, there are numerous advantages of using invertebrate animal models: less space and small equipment are enough for taking care of a large number of animals and thus are cost-effective, they can be easily handled, and many biological processes and genes are conserved between mammals and invertebrates. Today, many invertebrates have been used as animal models, but silkworms have many beneficial traits compared to mammals as well as other insects. In a Genome Pharmaceutical Institute's study, we were able to achieve a lot making use of silkworms as model animals. We would like to suggest that pharmaceutical companies and institutes consider the use of the silkworm as a model animal which is efficacious both for financial value by cost cutting and ethical aspects in animals' welfare.

  17. Research on non-destructive testing method of silkworm cocoons based on image processing technology

    NASA Astrophysics Data System (ADS)

    Gan, Yong; Kong, Qing-hua; Wei, Li-fu

    2008-03-01

    The major studied in this dissertation is the non-destructive testing method of silkworm cocoon's quality, based on the digital image processing and photoelectricity technology. Through the images collection and the data analysis, procession and calculation of the tested silkworm cocoons with the non-destructive testing technology, internet applications automatically reckon all items of the classification indexes. Finally we can conclude the classification result and the purchase price of the silkworm cocoons. According to the domestic classification standard of the silkworm cocoons, the author investigates various testing methods of silkworm cocoons which are used or have been explored at present, and devices a non-destructive testing scheme of the silkworm cocoons based on the digital image processing and photoelectricity technology. They are dissertated about the project design of the experiment. The precisions of all the implements are demonstrated. I establish Manifold mathematic models, compare them with each other and analyze the precision with technology of databank to get the best mathematic model to figure out the weight of the dried silkworm cocoon shells. The classification methods of all the complementary items are designed well and truly. The testing method has less error and reaches an advanced level of the present domestic non-destructive testing technology of the silkworm cocoons.

  18. Lactic acid bacteria activating innate immunity improve survival in bacterial infection model of silkworm.

    PubMed

    Nishida, Satoshi; Ono, Yasuo; Sekimizu, Kazuhisa

    2016-02-01

    Lactic acid bacteria (LAB) have been thought to be helpful for human heath in the gut as probiotics. It recently was noted that activity of LAB stimulating immune systems is important. Innate immune systems are conserved in mammals and insects. Silkworm has innate immunity in response to microbes. Microbe-associated molecular pattern (ex. peptidoglycan and β-glucan) induces a muscle contraction of silkworm larva. In this study, we established an efficient method to isolate lactic acid bacteria derived from natural products. We selected a highly active LAB to activate the innate immunity in silkworm by using the silkworm muscle contraction assay, as well. The assay revealed that Lactococcus lactis 11/19-B1 was highly active on the stimulation of the innate immunity in silkworm. L. lactis 11/19-B1 solely fermented milk with casamino acid and glucose. This strain would be a starter strain to make yogurt. Compared to commercially available yogurt LAB, L. lactis 11/19-B1 has higher activity on silkworm contraction. Silkworm normally ingested an artificial diet mixed with L. lactis 11/19-B1 or a yogurt fermented with L. lactis 11/19-B1. Interestingly, silkworms that ingested the LAB showed tolerance against the pathogenicity of Pseudomonas aeruginosa. These data suggest that Lactococcus lactis 11/19-B1 would be expected to be useful for making yogurt and probiotics to activate innate immunity. PMID:26971556

  19. Effect of Vitamin A on Listeria monocytogenes Infection in a Silkworm Model

    PubMed Central

    Watanabe, Kenta; Shimizu, Takashi; Watarai, Masahisa

    2016-01-01

    Insect infection models have been used increasingly to study various pathogenic agents in evaluations of pathogenicity and drug efficacy. In this study, we demonstrated that larvae of the silkworm Bombyx mori are useful for studying Listeria monocytogenes infections in insects. Infection with the L. monocytogenes wild-type strain induced silkworm death. Infection by a listeriolysin O (LLO) deletion mutant also induced silkworm death, but the bacterial numbers in silkworms were lower than those of the wild-type strain. Intracellular growth was observed when the silkworm ovary-derived cell line BmN4 was infected with the wild-type strain. Explosive replication was not observed in BmN4 cells infected with the LLO mutant and the bacterial numbers of the LLO mutant were lower than those of the wild-type strain. Pretreatment with vitamin A did not affect silkworm mortality after bacterial infection, but the efficiency of infecting the hemocytes and BmN4 cells was decreased with vitamin A treatment. Our results indicate that silkworm larvae are a useful insect infection model for L. monocytogenes and that vitamin A has protective effects against bacterial infection in silkworms. PMID:27669511

  20. Highly accurate and fast optical penetration-based silkworm gender separation system

    NASA Astrophysics Data System (ADS)

    Kamtongdee, Chakkrit; Sumriddetchkajorn, Sarun; Chanhorm, Sataporn

    2015-07-01

    Based on our research work in the last five years, this paper highlights our innovative optical sensing system that can identify and separate silkworm gender highly suitable for sericulture industry. The key idea relies on our proposed optical penetration concepts and once combined with simple image processing operations leads to high accuracy in identifying of silkworm gender. Inside the system, there are electronic and mechanical parts that assist in controlling the overall system operation, processing the optical signal, and separating the female from male silkworm pupae. With current system performance, we achieve a very highly accurate more than 95% in identifying gender of silkworm pupae with an average system operational speed of 30 silkworm pupae/minute. Three of our systems are already in operation at Thailand's Queen Sirikit Sericulture Centers.

  1. Symbiosis with Francisella tularensis provides resistance to pathogens in the silkworm.

    PubMed

    Suzuki, Jin; Uda, Akihiko; Watanabe, Kenta; Shimizu, Takashi; Watarai, Masahisa

    2016-01-01

    Francisella tularensis, the causative agent of tularemia, is a highly virulent facultative intracellular pathogen found in a wide range of animals, including arthropods, and environments. This bacterium has been known for over 100 years, but the lifestyle of F. tularensis in natural reservoirs remains largely unknown. Thus, we established a novel natural host model for F. tularensis using the silkworm (Bombyx mori), which is an insect model for infection by pathogens. F. tularensis established a symbiosis with silkworms, and bacteria were observed in the hemolymph. After infection with F. tularensis, the induction of melanization and nodulation, which are immune responses to bacterial infection, were inhibited in silkworms. Pre-inoculation of silkworms with F. tularensis enhanced the expression of antimicrobial peptides and resistance to infection by pathogenic bacteria. These results suggest that silkworms acquire host resistance via their symbiosis with F. tularensis, which may have important fitness benefits in natural reservoirs. PMID:27507264

  2. Symbiosis with Francisella tularensis provides resistance to pathogens in the silkworm

    PubMed Central

    Suzuki, Jin; Uda, Akihiko; Watanabe, Kenta; Shimizu, Takashi; Watarai, Masahisa

    2016-01-01

    Francisella tularensis, the causative agent of tularemia, is a highly virulent facultative intracellular pathogen found in a wide range of animals, including arthropods, and environments. This bacterium has been known for over 100 years, but the lifestyle of F. tularensis in natural reservoirs remains largely unknown. Thus, we established a novel natural host model for F. tularensis using the silkworm (Bombyx mori), which is an insect model for infection by pathogens. F. tularensis established a symbiosis with silkworms, and bacteria were observed in the hemolymph. After infection with F. tularensis, the induction of melanization and nodulation, which are immune responses to bacterial infection, were inhibited in silkworms. Pre-inoculation of silkworms with F. tularensis enhanced the expression of antimicrobial peptides and resistance to infection by pathogenic bacteria. These results suggest that silkworms acquire host resistance via their symbiosis with F. tularensis, which may have important fitness benefits in natural reservoirs. PMID:27507264

  3. Molecular characterization and phylogenetic analysis of the eukaryotic translation initiation factor 4A gene in Antheraea pernyi (Lepdoptera: Saturniidae).

    PubMed

    Chen, Miao-Miao; Liu, Yan-Qun; Li, Yan; Yao, Rui; Chen, Mo; Xia, Run-Xi; Li, Qun; Qin, Li

    2014-01-01

    Eukaryotic initiation factor 4A (eIF-4A) is an essential component for protein translation in eukaryotes. The eIF-4A gene (ApeIF-4A) was isolated and characterized from Antheraea pernyi (Guérin-Méneville) (Lepidoptera: Saturniidae). The obtained cDNA sequence was 1,435-bp long with an open reading frame of 1,266 bp encoding 421 amino acids. The predicted amino acid sequence shared several conserved features as found in known eIF-4As and revealed 74 and 78% identities with eIF-4As of Homo sapiens L. and Drosophila melanogaster (Meigen), respectively. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that ApeIF-4A was transcribed at four developmental stages and in all tissues tested, suggesting that it plays an important role in development of A. pernyi. Homologous alignment suggested that eIF-4As are highly conserved throughout evolution of eukaryote organisms. Phylogenetic trees based on the amino acid and nucleotide sequences of eIF-4A demonstrated a similar topology with the classical systematics, suggesting that it has the potential value in phylogenetic inference of eukaryotes.

  4. Molecular characterization of genome segment 2 encoding RNA dependent RNA polymerase of Antheraea mylitta cytoplasmic polyhedrosis virus

    SciTech Connect

    Ghorai, Suvankar; Chakrabarti, Mrinmay; Roy, Sobhan; Chavali, Venkata Ramana Murthy; Bagchi, Abhisek; Ghosh, Ananta Kumar

    2010-08-15

    Genome segment 2 (S2) from Antheraea mylitta cypovirus (AmCPV) was converted into cDNA, cloned and sequenced. S2 consisted of 3798 nucleotides with a long ORF encoding a 1116 amino acid long protein (123 kDa). BLAST and phylogenetic analysis showed 29% sequence identity and close relatedness of AmCPV S2 with RNA dependent RNA polymerase (RdRp) of other insect cypoviruses, suggesting a common origin of all insect cypoviruses. The ORF of S2 was expressed as 123 kDa soluble His-tagged fusion protein in insect cells via baculovirus recombinants which exhibited RdRp activity in an in vitro RNA polymerase assay without any intrinsic terminal transferase activity. Maximum activity was observed at 37 deg. C at pH 6.0 in the presence of 3 mM MgCl{sub 2.} Site directed mutagenesis confirmed the importance of the conserved GDD motif. This is the first report of functional characterization of a cypoviral RdRp which may lead to the development of anti-viral agents.

  5. Microarray analysis of New Green Cocoon associated genes in silkworm, Bombyx mori.

    PubMed

    Lu, Ya-Ru; He, Song-Zhen; Tong, Xiao-Ling; Han, Min-Jin; Li, Chun-Lin; Li, Zhi-Quan; Dai, Fang-Yin

    2016-06-01

    Green cocoons in silkworm, Bombyx mori, are caused by flavonoids accumulation in the silk proteins, fibroin and sericin. Despite the economic value of natural green cocoon and medical value of flavonoids, there is limited understanding of the molecular mechanism regulating flavonoids uptake in silkworm, which is tightly associated with the trait of green cocoon. The purpose of this study is to perform a comprehensive analysis to understand the molecular mechanisms of flavonoids uptake in silkworm based on microarray analyses. The study subject was the New Green Cocoon from the silkworm strains, G200 and N100, a new spontaneous dominant green cocoon trait identified in the 2000s. The genes regulating this trait are independent of other green cocoon genes previously reported. Genome-wide gene expression was compared between the New Green Cocoon producing silkworm strains, G200 and N100, and the control sample, which is the white cocoon producing strain 872B. Among these strains, N100 and 872B are near-isogenic lines. The results showed that 130 genes have consistently changing expression patterns in the green cocoon strains when compared with the white cocoon strain. Among these, we focused on the genes related to flavonoids metabolism and absorption, such as sugar transporter genes and UDP-glucosyltransferase genes. Based on our findings, we propose the potential mechanisms for flavonoids absorption and metabolism in silkworm. Our results imply that silkworm might be used as an underlying model for flavonoids in pharmaceutical research.

  6. Biological Effects of Low Energy Ar+ Ion Bombardment on Silkworm Eggs: a Novel Animal Model

    NASA Astrophysics Data System (ADS)

    Xu, Jiaping; Wu, Yuejin; Liu, Xuelan; Yuan, Hang; Yu, Zengliang

    2009-06-01

    In this study, we found for the first time that silkworm eggs were able to survive in vacuum for a long period of time. Subsequently, low energy Ar+ ions with different energies and fluences were used to bombard silkworm eggs so as to explore the resulting biological effects. Results showed that (i) the exposure of silkworm eggs to vacuum within 10 min did not cause significant impact on the hatching rates, while the irradiation of silkworm eggs by Ar+ ions of 25 keV or 30 keV with fluences ranging from 2.6×2.6 × 1015 ion/cm2 to 8×2.6 × 1015 ion/cm2 caused a significant impact on the hatching rates, and the hatching rates decreased with the increase in the fluence and energy level; (ii) the irradiation of silkworm eggs by Ar+ ions of 30 keV with a fluence of 8×2.6 × 1015 ion/cm2 or 9×2.6 × 1015 ion/cm2 resulted in a noticeable etching on the egg shell surface which could be observed by a scanning electron microscope; and (iii) the irradiation of silkworm eggs by Ar+ ions of 30 keV with a fluence of 9×2.6 × 1015 ion/cm2 generated several mutant phenotypes which were observed in the 5th instar silkworms and a moth.

  7. [Effects of silkworm pupa oil on serum lipids level and platelet function in rats].

    PubMed

    Yang, Xuefeng; Huang, Lianzhen; Hu, Jianping; Li, Tao

    2002-08-01

    To observe the effects of silkworm pupa oil on serum lipids level and platelet function in rats, according to serum TG, TC level, 40 male Wistar rats are divided into four groups (normal control group, high fat control group, silkworm pupa oil group and silkworm pupa oil + VE group). The rats are fed different diets and six weeks later, serum lipids level and platelet function are measured. The results show that (1) Compared with high fat control group, serum TC, TG, LDL-C level, AI value, Platelet aggregability, plasma TXB2 level and T/P ratio decrease significantly while HDL-C level and 6-k-PGF1 level increase in silkworm pupa oil group; (2) Serum TC, LDL-C level, T/P ratio and platelet aggregability are significantly lower in silkworm pupa oil + VE group than in silkworm pupa oil group. It is suggested that silkworm pupa oil rich in alpha-linolenic acid can reduce serum lipids level and inhibit platelet aggregation, which is more effective with the supplementation with VE.

  8. BmTEdb: a collective database of transposable elements in the silkworm genome.

    PubMed

    Xu, Hong-En; Zhang, Hua-Hao; Xia, Tian; Han, Min-Jin; Shen, Yi-Hong; Zhang, Ze

    2013-01-01

    The silkworm, Bombyx mori, is one of the major insect model organisms, and its draft and fine genome sequences became available in 2004 and 2008, respectively. Transposable elements (TEs) constitute ~40% of the silkworm genome. To better understand the roles of TEs in organization, structure and evolution of the silkworm genome, we used a combination of de novo, structure-based and homology-based approaches for identification of the silkworm TEs and identified 1308 silkworm TE families. These TE families and their classification information were organized into a comprehensive and easy-to-use web-based database, BmTEdb. Users are entitled to browse, search and download the sequences in the database. Sequence analyses such as BLAST, HMMER and EMBOSS GetORF were also provided in BmTEdb. This database will facilitate studies for the silkworm genomics, the TE functions in the silkworm and the comparative analysis of the insect TEs. Database URL: http://gene.cqu.edu.cn/BmTEdb/.

  9. Silkworm cocoons inspire models for random fiber and particulate composites

    NASA Astrophysics Data System (ADS)

    Chen, Fujia; Porter, David; Vollrath, Fritz

    2010-10-01

    The bioengineering design principles evolved in silkworm cocoons make them ideal natural prototypes and models for structural composites. Cocoons depend for their stiffness and strength on the connectivity of bonding between their constituent materials of silk fibers and sericin binder. Strain-activated mechanisms for loss of bonding connectivity in cocoons can be translated directly into a surprisingly simple yet universal set of physically realistic as well as predictive quantitative structure-property relations for a wide range of technologically important fiber and particulate composite materials.

  10. Silkworm cocoons inspire models for random fiber and particulate composites.

    PubMed

    Chen, Fujia; Porter, David; Vollrath, Fritz

    2010-10-01

    The bioengineering design principles evolved in silkworm cocoons make them ideal natural prototypes and models for structural composites. Cocoons depend for their stiffness and strength on the connectivity of bonding between their constituent materials of silk fibers and sericin binder. Strain-activated mechanisms for loss of bonding connectivity in cocoons can be translated directly into a surprisingly simple yet universal set of physically realistic as well as predictive quantitative structure-property relations for a wide range of technologically important fiber and particulate composite materials.

  11. Silkworm cocoons inspire models for random fiber and particulate composites

    SciTech Connect

    Chen Fujia; Porter, David; Vollrath, Fritz

    2010-10-15

    The bioengineering design principles evolved in silkworm cocoons make them ideal natural prototypes and models for structural composites. Cocoons depend for their stiffness and strength on the connectivity of bonding between their constituent materials of silk fibers and sericin binder. Strain-activated mechanisms for loss of bonding connectivity in cocoons can be translated directly into a surprisingly simple yet universal set of physically realistic as well as predictive quantitative structure-property relations for a wide range of technologically important fiber and particulate composite materials.

  12. Transcriptome analysis of the silkworm (Bombyx mori) by high-throughput RNA sequencing.

    PubMed

    Li, Yinü; Wang, Guozeng; Tian, Jian; Liu, Huifen; Yang, Huipeng; Yi, Yongzhu; Wang, Jinhui; Shi, Xiaofeng; Jiang, Feng; Yao, Bin; Zhang, Zhifang

    2012-01-01

    The domestic silkworm, Bombyx mori, is a model insect with important economic value for silk production that also acts as a bioreactor for biomaterial production. The functional complexity of the silkworm transcriptome has not yet been fully elucidated, although genomic sequencing and other tools have been widely used in its study. We explored the transcriptome of silkworm at different developmental stages using high-throughput paired-end RNA sequencing. A total of about 3.3 gigabases (Gb) of sequence was obtained, representing about a 7-fold coverage of the B. mori genome. From the reads that were mapped to the genome sequence; 23,461 transcripts were obtained, 5,428 of them were novel. Of the 14,623 predicted protein-coding genes in the silkworm genome database, 11,884 of them were found to be expressed in the silkworm transcriptome, giving a coverage of 81.3%. A total of 13,195 new exons were detected, of which, 5,911 were found in the annotated genes in the Silkworm Genome Database (SilkDB). An analysis of alternative splicing in the transcriptome revealed that 3,247 genes had undergone alternative splicing. To help with the data analysis, a transcriptome database that integrates our transcriptome data with the silkworm genome data was constructed and is publicly available at http://124.17.27.136/gbrowse2/. To our knowledge, this is the first study to elucidate the silkworm transcriptome using high-throughput RNA sequencing technology. Our data indicate that the transcriptome of silkworm is much more complex than previously anticipated. This work provides tools and resources for the identification of new functional elements and paves the way for future functional genomics studies. PMID:22928022

  13. Production of scFv-Conjugated Affinity Silk Powder by Transgenic Silkworm Technology

    PubMed Central

    Sato, Mitsuru; Kojima, Katsura; Sakuma, Chisato; Murakami, Maria; Aratani, Eriko; Takenouchi, Takato; Tamada, Yasushi; Kitani, Hiroshi

    2012-01-01

    Bombyx mori (silkworm) silk proteins are being utilized as unique biomaterials for medical applications. Chemical modification or post-conjugation of bioactive ligands expand the applicability of silk proteins; however, the processes are elaborate and costly. In this study, we used transgenic silkworm technology to develop single-chain variable fragment (scFv)-conjugated silk fibroin. The cocoons of the transgenic silkworm contain fibroin L-chain linked with scFv as a fusion protein. After dissolving the cocoons in lithium bromide, the silk solution was dialyzed, concentrated, freeze-dried, and crushed into powder. Immunoprecipitation analyses demonstrate that the scFv domain retains its specific binding activity to the target molecule after multiple processing steps. These results strongly suggest the promise of scFv-conjugated silk fibroin as an alternative affinity reagent, which can be manufactured using transgenic silkworm technology at lower cost than traditional affinity carriers. PMID:22496833

  14. Characterization and Comparison of Serratia Marcescens Isolated from Edible Cactus and from Silkworm for Virulence Potential and Chitosan Susceptibility

    PubMed Central

    Li, Bin; Yu, Rongrong; Liu, Baoping; Tang, Qiaomei; Zhang, Guoqing; Wang, Yanli; Xie, Guanlin; Sun, Guochang

    2011-01-01

    Representative strains of Serratia marcescens from an edible cactus plant and silkworms were characterized and a comparison based on their cellular fatty acid composition, 16S rRNA and groE gene sequence analysis as well as silkworm virulence and chitosan susceptibility was carried out. Results from this study indicate that there are no significant differences between the phenotypic and molecular characterization, virulence and chitosan susceptibility of the S. marcescens strains from the cactus plant and silkworms. Silkworms inoculated with S. marcescens from either plant or silkworm resulted in nearly 100% mortality. Chitosan solution exhibited strong antibacterial activity against S. marcescens. This activity increased with the increase of chitosan concentration and incubation time regardless of the strain source. Also, the results indicate that the plant associated S. marcescens maybe plays a possible role in the contamination of humans and animals, in particular silkworms, while chitosan showed a potential to control the contamination caused by S. marcescens. PMID:24031610

  15. Characterization and comparison of serratia marcescens isolated from edible cactus and from silkworm for virulence potential and chitosan susceptibility.

    PubMed

    Li, Bin; Yu, Rongrong; Liu, Baoping; Tang, Qiaomei; Zhang, Guoqing; Wang, Yanli; Xie, Guanlin; Sun, Guochang

    2011-01-01

    Representative strains of Serratia marcescens from an edible cactus plant and silkworms were characterized and a comparison based on their cellular fatty acid composition, 16S rRNA and groE gene sequence analysis as well as silkworm virulence and chitosan susceptibility was carried out. Results from this study indicate that there are no significant differences between the phenotypic and molecular characterization, virulence and chitosan susceptibility of the S. marcescens strains from the cactus plant and silkworms. Silkworms inoculated with S. marcescens from either plant or silkworm resulted in nearly 100% mortality. Chitosan solution exhibited strong antibacterial activity against S. marcescens. This activity increased with the increase of chitosan concentration and incubation time regardless of the strain source. Also, the results indicate that the plant associated S. marcescens maybe plays a possible role in the contamination of humans and animals, in particular silkworms, while chitosan showed a potential to control the contamination caused by S. marcescens.

  16. Analysis of silkworm gut microflora in the Bioregenerative Life Support System

    NASA Astrophysics Data System (ADS)

    Liang, Xue; Liu, lh64. Hong

    2012-07-01

    Silkworm (Bombyx mori L) has advantages in the nutritional composition, growth characteristics and other factors, it is regarded as animal protein source for astronauts in the Bioregenerative Life Support System (BLSS).Due to the features of BLSS, silkworm breeding way is different from the conventional one (mulberry leaves throughout five instars): they were fed with mulberry and lettuce leaves during the 1st-3rd instars and 4th -5th instars, respectively. As the lettuce stem can be eaten by astronauts, the leaves not favored by humans can be insect's foodstuff. Therefore, it is necessary to investigate the gut microbial composition, the type of dominant bacteria of silkworm raised with this way and the differences from the conventional breeding method, so as to reduce the mortality rate caused by the foodstuff change and to provide more animal protein for astronauts. In this study, 16srDNA sequencing, phylogenetic analysis and denaturing gradient gel electrophoresis method were used to analyze the silkworm gut microbial flora under two breeding manners. The results show that conventional and BLSS breeding way have six dominant bacteria in common: Clostridium, Enterococcus, Bacteroides, Chryseobacterium, Parabacteroides, Paenibacillus. We also found Escherichia, Janthinobacterium, Sedimentibacter, Streptococcus, Bacillus, Arcobacter, Rothia, Polaribacter and Acinetobacter, Anaerofilum, Rummeliibacillus, Anaeroplasma, Serratia in the ground conventional and BLSS special breeding way, respectively. Changing the foodstuff of silkworm leads to the dynamic alteration of gut microbial. Dominant bacteria of the two breeding ways have diversities from each other. The ground conventional breeding way has more abundant bacteria than the BLSS one. Due to the lettuce leaves have replaced mulberry leaves at the beginning of the silkworm 4th instar, some silkworms can not survive without the bacteria that digest and absorb lettuce leaves. We suggest those dominant bacteria

  17. Mechanism of enhanced Bombyx mori nucleopolyhedrovirus-resistance by titanium dioxide nanoparticles in silkworm.

    PubMed

    Xu, Kaizun; Li, Fanchi; Ma, Lie; Wang, Binbin; Zhang, Hua; Ni, Min; Hong, Fashui; Shen, Weide; Li, Bing

    2015-01-01

    The infection of Bombyx mori nucleopolyhedrovirus (BmNPV) in silkworms is often lethal. It is difficult to prevent, and its lethality is correlated with both viral particle characteristics and silkworm strains. Low doses of titanium dioxide nanoparticles (TiO2 NPs) can promote silkworm growth and improve its resistance to organophosphate pesticides. In this study, TiO2 NPs' effect on BmNPV resistance was investigated by analyzing the characteristics of BmNPV proliferation and transcriptional differences in silkworm midgut and the transcriptional changes of immunity related genes after feeding with TiO2 NPs. We found that low doses of TiO2 NPs improved the resistance of silkworm against BmNPV by 14.88-fold, with the mortalities of the experimental group and control group being 0.56% and 8.33% at 144 h, respectively. The proliferation of BmNPV in the midgut was significantly increased 72 h after infection in both experimental and control groups; the control group reached the peak at 120 h, while the experimental group took 24 more hours to reach the maximal value that was 12.63 times lower than the control, indicating that TiO2 NPs can inhibit BmNPV proliferation in the midgut. Consistently, the expression of the BmNPV-resistant gene Bmlipase-1 had the same increase pattern as the proliferation changes. Immune signaling pathway analysis revealed that TiO2 NPs inhibited the proliferation of silkworm BmNPV to reduce the activation levels of janus kinase/signal transducer and activator of transcription (JAK/STAT) and phosphatidylinositol 3-kinase (PI3K)-Akt signaling pathway, while promoting the expression of Bmakt to improve the immunity. Overall, our results demonstrate that TiO2 NPs increase silkworm resistance against BmNPV by inhibiting virus proliferation and improving immunity in silkworms.

  18. Vertebrate estrogen regulates the development of female characteristics in silkworm, Bombyx mori.

    PubMed

    Shen, Guanwang; Lin, Ying; Yang, Congwen; Xing, Runmiao; Zhang, Haiyan; Chen, Enxiang; Han, Chaoshan; Liu, Hongling; Zhang, Weiwei; Xia, Qingyou

    2015-01-01

    The vertebrate estrogens include 17-β-estradiol (E2), which has an analog in silkworm ovaries. In this study, the Bombyx mori vitellogenin gene (BmVg) was used as a biomarker to analyze the function of the E2 in silkworm. In most oviparous animals, Vg has female-specific expression. However, BmVg expression was also detected in B. mori males. Stage specific fluctuation of BmVg expression was similar in males and females, but expression levels in males were lower than in females. E2 treatment by injection or feeding of male larvae in the final instar stage induced and stimulated male BmVg transcription and protein synthesis. When silkworm ovary primordia were transplanted into males, BmVg was induced in male fat bodies. Transplanted ovaries primordia were also able to develop into ovaries and produce mature eggs. When females were treated with E2 promoted BmVg/BmVn protein accumulation in hemolymph, ovaries and eggs. However, BmVg transcription was decreased in female fat bodies. An E2 analog was identified in the hemolymph of day 3 wandering silkworms using high-performance liquid chromatography. Estradiol titers from fifth late-instar larvae to pupal stage were determined by enzyme-linked immunosorbent assay. The results suggested that silkworms synthesized a vertebrate E2 analog. This study found that E2 promoted the synthesis of BmVg, a female typical protein in silkworms.

  19. The profiles of red fluorescent proteins with antinucleopolyhedrovirus activity in races of the silkworm Bombyx mori.

    PubMed

    Sunagar, Santosh G; Savanurmath, Chandrashekhar J; Hinchigeri, Shivayogeppa B

    2011-12-01

    Partially purified red fluorescent proteins (RFPs) secured from the gut juice of 5th-instar multivoltine and bivoltine silkworm races were observed as several bands in electrophoretograms and chromatographic eluates. Interestingly, different races of silkworms had varying numbers of fluorescent protein bands: 11 in Pure Mysore (resistant), 11 in Nistari (resistant), 4 in CSR(2) (moderately susceptible) and 1 in NB(4)D(2) (highly susceptible). Bioassay experiments indicated that the fluorescent bands had antinucleopolyhedrovirus (antiNPV) activity. The molar extinction coefficients and fluorescence quantum yields of all RFPs were estimated. The purified tetrapyrroles were characterized by UV-visible absorption and fluorescence spectral analyses. All tetrapyrrole moieties associated with RFPs were found to be different and characteristic of the fluorescent bands. The resulting qualitative and quantitative differences among the individual RFPs from various races of silkworm were related to the susceptibilities of the silkworms to the viral disease. Moreover, light was found to be essential for the synthesis of RFPs, and, therefore, the role of light in the synthesis of RFPs was evaluated. Thus, this work may elucidate the process of RFP synthesis in silkworm, which may be used as a biomarker to measure the degree of susceptibility of silkworm races to NPV. Therefore, the characteristic band pattern may be used as an indicator to define the relative resistance of a race towards the specific virus.

  20. Rhodiola rosea extends lifespan and improves stress tolerance in silkworm, Bombyx mori.

    PubMed

    Chen, Cong; Song, Jiangbo; Chen, Min; Li, Zhiquan; Tong, Xiaoling; Hu, Hai; Xiang, Zhonghuai; Lu, Cheng; Dai, Fangyin

    2016-04-01

    The root of Rhodiola rosea is widely used in Traditional Chinese Medicine. The extract from R. rosea is reported to extend the lifespan of yeast, nematode, and fruit fly. However, the molecular mechanism is not fully understood. Here, we tested whether R. rosea extends the lifespan of the silkworm. An aqueous extract of R. rosea significantly prolonged the lifespan of the silkworm, without affecting its daily food intake, body weight, or fecundity, suggesting that R. rosea did not exhibit obvious side effects. Rhodiola rosea extract also enhanced the stress resistance in the silkworm, against heat stress (37 °C) and starvation. The R. rosea extract increased the activity of the major antioxidant enzymes, glutathione S-transferase and catalase, and altered the content of glutathione and malondialdehyde. Rhodiola rosea increased the expression of BmFoxO, which is a downstream regulator of insulin/IGF-1 signaling (IIS) pathway in the silkworm. Our results showed that R. rosea extends lifespan, in which IIS pathway might be involved, and enhances stress resistance in the silkworm. Thus, the silkworm might be used as a novel animal model for lifespan study and efficacy evaluation of Traditional Chinese Medicines.

  1. Improvement of light penetration based silkworm gender identification with confined regions of interest

    NASA Astrophysics Data System (ADS)

    Kamtongdee, Chakkrit; Sumriddetchkajorn, Sarun; Sa-ngiamsak, Chiranut

    2013-06-01

    Based on our previous work on light penetration-based silkworm gender identification, we find that unwanted optical noises scattering from the surrounding area near the silkworm pupa and the transparent support are sometimes analyzed and misinterpreted leading to incorrect silkworm gender identification. To alleviate this issue, we place a small rectangular hole on a transparent support so that it not only helps the user precisely place the silkworm pupa but also functions as a region of interest (ROI) for blocking unwanted optical noises and for roughly locating the abdomen region in the image for ease of image processing. Apart from the external ROI, we also assign a smaller ROI inside the image in order to remove strong scattering light from all edges of the external ROI and at the same time speed up our image processing operations. With only the external ROI in function, our experiment shows a measured 86% total accuracy in identifying gender of 120 silkworm pupae with a measured average processing time of 38 ms. Combining the external ROI and the image ROI together revamps the total accuracy in identifying the silkworm gender to 95% with a measured faster 18 ms processing time.

  2. Metabolomics Analysis of the Larval Head of the Silkworm, Bombyx mori

    PubMed Central

    Li, Yi; Wang, Xin; Chen, Quanmei; Hou, Yong; Xia, Qingyou; Zhao, Ping

    2016-01-01

    The head, which performs many biological functions, is the most complicated structure of an insect. Development, locomotor behavior, food intake, environmental sensing, and signal transduction are all controlled by the insect’s head. As a well-studied insect in Lepidoptera, the silkworm head has an additional function of spinning silk fibers. To understand which molecules are involved in these physiological activities, we performed a metabolomics analysis of silkworm heads. By integrating GC-MS and LC-MS/MS, 90 metabolites were identified in the larval heads of silkworms. These were classified into 13 categories, including amino acids, sugars, organic acids, nucleotides, alcohols, and fatty acids. Informatics analysis revealed that these metabolites are involved in cellular processes, environmental information processing, genetic information processing, human diseases, metabolism, organismal systems, and other pathways. The identified metabolites and pathways are involved in biological processes such as signal transduction, carbohydrate metabolism, endocrine activities, and sensory activities; reflecting the functions of various organs in silkworm heads. Thus, our findings provide references which elucidate the potential functions of the silkworm head and will be of great value for the metabolomics research of silkworms and other insects. PMID:27657048

  3. Transcriptome Analysis of Silkworm, Bombyx mori, during Early Response to Beauveria bassiana Challenges

    PubMed Central

    Hou, Chengxiang; Qin, Guangxing; Liu, Ting; Geng, Tao; Gao, Kun; Pan, Zhonghua; Qian, Heying; Guo, Xijie

    2014-01-01

    Host–pathogen interactions are complex processes and it is a central challenge to reveal these interactions. Fungal infection of silkworm, Bombyx mori, may induce a variety of responsive reaction. However, little is known about the molecular mechanism of silkworm immune response against the fungal infection. To obtain an overview of the interaction between silkworm and an entomopathogenic fungus Beauveria bassiana, Digital Gene Expression profiling, a tag based high-throughput transcriptome sequencing method, was employed to screen and identify differentially expressed genes (DEGs, FDR≤0.001, ∣log2ratio∣≥1) of silkworm larvae during early response against B. bassiana infection. Total 1430 DEGs including 960 up-regulated and 470 down-regulated ones were identified, of which 627 DEGs can be classified into GO categories by Gene Ontology (GO) analysis. KEGG pathways analysis of these DEGs suggested that many biological processes, such as defense and response, signal transduction, phagocytosis, regulation of gene expression, RNA splicing, biosynthesis and metabolism, protein transport etc. were involved in the interaction between the silkworm and B. bassiana. A number of differentially expressed fungal genes were also identified by mapping the sequencing tags to B. bassiana genome. These results provided new insights to the molecular mechanism of silkworm immune response to B. bassiana infection. PMID:24618587

  4. Rhodiola rosea extends lifespan and improves stress tolerance in silkworm, Bombyx mori.

    PubMed

    Chen, Cong; Song, Jiangbo; Chen, Min; Li, Zhiquan; Tong, Xiaoling; Hu, Hai; Xiang, Zhonghuai; Lu, Cheng; Dai, Fangyin

    2016-04-01

    The root of Rhodiola rosea is widely used in Traditional Chinese Medicine. The extract from R. rosea is reported to extend the lifespan of yeast, nematode, and fruit fly. However, the molecular mechanism is not fully understood. Here, we tested whether R. rosea extends the lifespan of the silkworm. An aqueous extract of R. rosea significantly prolonged the lifespan of the silkworm, without affecting its daily food intake, body weight, or fecundity, suggesting that R. rosea did not exhibit obvious side effects. Rhodiola rosea extract also enhanced the stress resistance in the silkworm, against heat stress (37 °C) and starvation. The R. rosea extract increased the activity of the major antioxidant enzymes, glutathione S-transferase and catalase, and altered the content of glutathione and malondialdehyde. Rhodiola rosea increased the expression of BmFoxO, which is a downstream regulator of insulin/IGF-1 signaling (IIS) pathway in the silkworm. Our results showed that R. rosea extends lifespan, in which IIS pathway might be involved, and enhances stress resistance in the silkworm. Thus, the silkworm might be used as a novel animal model for lifespan study and efficacy evaluation of Traditional Chinese Medicines. PMID:26497336

  5. Proteomic and immunological identification of two new allergens from silkworm (Bombyx mori L.) pupae

    PubMed Central

    Zhao, Xiangjie; Li, Lin; Kuang, Zheshi; Luo, Guoqing; Li, BING

    2015-01-01

    This study explored food allergy caused by eating silkworm (Bombyx mori L.) pupae, a traditionally accepted food and animal feed in East and Southeast Asia, and identified two new allergens by proteomic and immunological methods. Proteins isolated from silkworm pupae were separated by two-dimensional gel electrophoresis (2-DE); pooled sera from patients allergic to silkworm pupa proteins were used to detect immunoglobulin E (IgE)-binding proteins by western blotting, and allergens specific for silkworm pupa consumption-caused allergy were visualised with the ECL reagents. The selected allergen proteins were further identified by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) analysis. Finally, chitinase and paramyosin were identified as silkworm pupa proteins showing strong immunoglobulin (IgE)-binding reaction. Analysis of the sequence homology of the two proteins using the AllergenOnline database indicated that chitinase and paramyosin shared 24.8% and 62.8% sequence homology with known allergens Der f 18 (Dermatophagoides farinae) and Der p 11 (Dermatophagoides pteronyssinus), respectively. Our results shed light on the understanding and treatment of silkworm pupa allergy. PMID:26155181

  6. Cadmium transfer and detoxification mechanisms in a soil-mulberry-silkworm system: phytoremediation potential.

    PubMed

    Zhou, Lingyun; Zhao, Ye; Wang, Shuifeng

    2015-11-01

    Phytoremediation has been proven to be an environmentally sound alternative for the recovery of contaminated soils, and the economic profit that comes along with the process might stimulate its field use. This study investigated cadmium (Cd) transfer and detoxification mechanisms in a soil-mulberry-silkworm system to estimate the suitability of the mulberry and silkworm as an alternative method for the remediation of Cd-polluted soil; it also explored the underlying mechanisms regulating the trophic transfer of Cd. The results show that both the mulberry and silkworm have high Cd tolerance. The transfer factor suggests that the mulberry has high potential for Cd extraction from polluted soil. The subcellular distribution and chemical forms of Cd in mulberry leaves show that cell wall deposition and vacuolar compartmentalization play important role in Cd tolerance. In the presence of increasing Cd concentrations in silkworm food, detoxification mechanisms (excretion and homeostasis) were activated so that excess Cd was excreted in fecal balls, and metallothionein levels in the mid-gut, the posterior of the silk gland, and the fat body of silkworms were enhanced. And, the Cd concentrations in silk are at a low level, ranging from 0.02 to 0.21 mg kg(-1). Therefore, these mechanisms of detoxification can regulate Cd trophic transfer, and mulberry planting and silkworm breeding has high phytoremediation potential for Cd-contaminated soil.

  7. An adaptive transposable element insertion in the regulatory region of the EO gene in the domesticated silkworm, Bombyx mori.

    PubMed

    Sun, Wei; Shen, Yi-Hong; Han, Min-Jin; Cao, Yun-Feng; Zhang, Ze

    2014-12-01

    Although there are many studies to show a key role of transposable elements (TEs) in adaptive evolution of higher organisms, little is known about the molecular mechanisms. In this study, we found that a partial TE (Taguchi) inserted in the cis-regulatory region of the silkworm ecdysone oxidase (EO) gene, which encodes a crucial enzyme to reduce the titer of molting hormone (20-hydroxyecdysone, 20E). The TE insertion occurred during domestication of silkworm and the frequency of the TE insertion in the domesticated silkworm (Bombyx mori) is high, 54.24%. The linkage disequilibrium in the TE inserted strains of the domesticated silkworm was elevated. Molecular population genetics analyses suggest that this TE insertion is adaptive for the domesticated silkworm. Luminescent reporter assay shows that the TE inserted in the cis-regulatory region of the EO gene functions as a 20E-induced enhancer of the gene expression. Further, phenotypic bioassay indicates that the silkworm with the TE insertion exhibited more stable developmental phenotype than the silkworm without the TE insertion when suffering from food shortage. Thus, the inserted TE in the cis-regulatory region of the EO gene increased developmental uniformity of silkworm individuals through regulating 20E metabolism, partially explaining transformation of a domestication developmental trait in the domesticated silkworm. Our results emphasize the exceptional role of gene expression regulation in developmental transition of domesticated animals.

  8. Comparing the rheology of native spider and silkworm spinning dope

    NASA Astrophysics Data System (ADS)

    Holland, C.; Terry, A. E.; Porter, D.; Vollrath, F.

    2006-11-01

    Silk production has evolved to be energetically efficient and functionally optimized, yielding a material that can outperform most industrial fibres, particularly in toughness. Spider silk has hitherto defied all attempts at reproduction, despite advances in our understanding of the molecular mechanisms behind its superb mechanical properties. Spun fibres, natural and man-made, rely on the extrusion process to facilitate molecular orientation and bonding. Hence a full understanding of the flow characteristics of native spinning feedstock (dope) will be essential to translate natural spinning to artificial silk production. Here we show remarkable similarity between the rheologies for native spider-dragline and silkworm-cocoon silk, despite their independent evolution and substantial differences in protein structure. Surprisingly, both dopes behave like typical polymer melts. This observation opens the door to using polymer theory to clarify our general understanding of natural silks, despite the many specializations found in different animal species.

  9. Proteome identification of the silkworm middle silk gland

    PubMed Central

    Li, Jian-ying; Ye, Lu-peng; Che, Jia-qian; Song, Jia; You, Zheng-ying; Wang, Shao-hua; Zhong, Bo-xiong

    2016-01-01

    To investigate the functional differentiation among the anterior (A), middle (M), and posterior (P) regions of silkworm middle silk gland (MSG), their proteomes were characterized by shotgun LC–MS/MS analysis with a LTQ-Orbitrap mass spectrometer. To get better proteome identification and quantification, triplicate replicates of mass spectrometry analysis were performed for each sample. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (Vizcaíno et al., 2014) [1] via the PRIDE partner repository (Vizcaino, 2013) [2] with the dataset identifier PXD003371. The peptide identifications that were further processed by PeptideProphet program in Trans-Proteomic Pipeline (TPP) after database search with Mascot software were also available in .XML format files. Data presented here are related to a research article published in Journal of Proteomics by Li et al. (2015) [3]. PMID:26937469

  10. Proteome identification of the silkworm middle silk gland.

    PubMed

    Li, Jian-Ying; Ye, Lu-Peng; Che, Jia-Qian; Song, Jia; You, Zheng-Ying; Wang, Shao-Hua; Zhong, Bo-Xiong

    2016-03-01

    To investigate the functional differentiation among the anterior (A), middle (M), and posterior (P) regions of silkworm middle silk gland (MSG), their proteomes were characterized by shotgun LC-MS/MS analysis with a LTQ-Orbitrap mass spectrometer. To get better proteome identification and quantification, triplicate replicates of mass spectrometry analysis were performed for each sample. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (Vizcaíno et al., 2014) [1] via the PRIDE partner repository (Vizcaino, 2013) [2] with the dataset identifier PXD003371. The peptide identifications that were further processed by PeptideProphet program in Trans-Proteomic Pipeline (TPP) after database search with Mascot software were also available in .XML format files. Data presented here are related to a research article published in Journal of Proteomics by Li et al. (2015) [3]. PMID:26937469

  11. N-acetyltransferase (nat) Is a Critical Conjunct of Photoperiodism between the Circadian System and Endocrine Axis in Antheraea pernyi

    PubMed Central

    Bembenek, Jadwiga; Hiragaki, Susumu; Suzuki, Takeshi; Takeda, Makio

    2014-01-01

    Since its discovery in 1923, the biology of photoperiodism remains a mystery in many ways. We sought the link connecting the circadian system to an endocrine switch, using Antheraea pernyi. PER-, CLK- and CYC-ir were co-expressed in two pairs of dorsolateral neurons of the protocerebrum, suggesting that these are the circadian neurons that also express melatonin-, NAT- and HIOMT-ir. The results suggest that a melatonin pathway is present in the circadian neurons. Melatonin receptor (MT2 or MEL-1B-R)-ir in PTTH-ir neurons juxtaposing clock neurons suggests that melatonin gates PTTH release. RIA showed a melatonin rhythm with a peak four hours after lights off in adult brain both under LD16∶8 (LD) and LD12∶12 (SD), and both the peak and the baseline levels were higher under LD than SD, suggesting a photoperiodic influence. When pupae in diapause were exposed to 10 cycles of LD, or stored at 4°C for 4 months under constant darkness, an increase of NAT activity was observed when PTTH released ecdysone. DNA sequence upstream of nat contained E-boxes to which CYC/CLK could bind, and nat transcription was turned off by clk or cyc dsRNA. dsRNANAT caused dysfunction of photoperiodism. dsRNAPER upregulated nat transcription as anticipated, based on findings in the Drosophila melanogaster circadian system. Transcription of nat, cyc and clk peaked at ZT12. RIA showed that dsRNANAT decreased melatonin while dsRNAPER increased melatonin. Thus nat, a clock controlled gene, is the critical link between the circadian clock and endocrine switch. MT-binding may release PTTH, resulting in termination of diapause. This study thus examined all of the basic functional units from the clock: a photoperiodic counter as an accumulator of mRNANAT, to endocrine switch for photoperiodism in A. pernyi showing this system is self-complete without additional device especially for photoperiodism. PMID:24667367

  12. Evaluation of General Toxicity and Genotoxicity of the Silkworm Extract Powder

    PubMed Central

    Heo, Hyun-Suk; Choi, Jae-Hun; Oh, Jung-Ja; Lee, Woo-Joo; Kim, Seong-Sook; Lee, Do-Hoon; Lee, Hyun-Kul; Song, Si-Whan; Kim, Kap-Ho; Choi, Yang-Kyu; Ryu, Kang-Sun

    2013-01-01

    The silkworm extract powder contain 1-deoxynojirimycin (DNJ), a potent α-glycosidase inhibitor, has therapeutic potency against diabetes mellitus. Therefore, natural products containing DNJ from mulberry leaves and silkworm are consumed as health functional food. The present study was performed to evaluate the safety of the silkworm extract powder, a health food which containing the DNJ. The repeated toxicity studies and gentic toxicity studies of the silkworm extract powder were performed to obtain the data for new functional food approval in MFDS. The safety was evaluated by a single-dose oral toxicity study and a 90 day repeated-dose oral toxicity study in Sprague-Dawley rats. The silkworm extract powder was also evaluated for its mutagenic potential in a battery of genetic toxicity test: in vitro bacterial reverse mutation assay, in vitro chromosomal aberration test, and in vivo mouse bone marrow micronucleus assay. The results of the genetic toxicology assays were negative in all of the assays. The approximate lethal dose in single oral dose toxicity study was considered to be higher than 5000 mg/kg in rats. In the 90 day study, the dose levels were wet at 0, 500, 1000, 2000 mg/kg/day, and 10 animals/sex/dose were treated with oral gavage. The parameters that were monitored were clinical signs, body weights, food and water consumptions, ophthalmic examination, urinalysis, hematology, serum biochemistry, necropsy findings, organ weights, and histopathological examination. No adverse effects were observed after the 90 day administration of the silkworm extract powder. The No-Observed-Adverse-Effect-Level (NOAEL) of silkworm extract powder in the 90 day study was 2000 mg/kg/day in both sexes, and no target organ was identified. PMID:24578797

  13. Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)

    PubMed Central

    Tada, Minoru; Tatematsu, Ken-Ichiro; Ishii-Watabe, Akiko; Harazono, Akira; Takakura, Daisuke; Hashii, Noritaka; Sezutsu, Hideki; Kawasaki, Nana

    2015-01-01

    In response to the successful use of monoclonal antibodies (mAbs) in the treatment of various diseases, systems for expressing recombinant mAbs using transgenic animals or plants have been widely developed. The silkworm (Bombyx mori) is a highly domesticated insect that has recently been used for the production of recombinant proteins. Because of their cost-effective breeding and relatively easy production scale-up, transgenic silkworms show great promise as a novel production system for mAbs. In this study, we established a transgenic silkworm stably expressing a human-mouse chimeric anti-CD20 mAb having the same amino acid sequence as rituximab, and compared its characteristics with rituximab produced by Chinese hamster ovary (CHO) cells (MabThera®). The anti-CD20 mAb produced in the transgenic silkworm showed a similar antigen-binding property, but stronger antibody-dependent cell-mediated cytotoxicity (ADCC) and weaker complement-dependent cytotoxicity (CDC) compared to MabThera. Post-translational modification analysis was performed by peptide mapping using liquid chromatography/mass spectrometry. There was a significant difference in the N-glycosylation profile between the CHO− and the silkworm-derived mAbs, but not in other post-translational modifications including oxidation and deamidation. The mass spectra of the N-glycosylated peptide revealed that the observed biological properties were attributable to the characteristic N-glycan structures of the anti-CD20 mAbs produced in the transgenic silkworms, i.e., the lack of the core-fucose and galactose at the non-reducing terminal. These results suggest that the transgenic silkworm may be a promising expression system for the tumor-targeting mAbs with higher ADCC activity. PMID:26261057

  14. Heme precursor injection is effective for Arthromyces ramosus peroxidase fusion protein production by a silkworm expression system.

    PubMed

    Hayashi, Kounosuke; Lee, Jae Man; Tomozoe, Yusuke; Kusakabe, Takahiro; Kamiya, Noriho

    2015-10-01

    Recombinant peroxidase from Arthromyces ramosus, fused with domains of antibody-binding proteins, was successfully obtained by a silkworm larvae expression system. The catalytic activity of the fusion peroxidase was increased 6-fold with the injection of 5-aminolevulinic acid into silkworm larvae as a heme precursor.

  15. Genome-wide identification and characterization of Fox genes in the silkworm, Bombyx mori.

    PubMed

    Song, JiangBo; Li, ZhiQuan; Tong, XiaoLing; Chen, Cong; Chen, Min; Meng, Gang; Chen, Peng; Li, ChunLin; Xin, YaQun; Gai, TingTing; Dai, FangYin; Lu, Cheng

    2015-09-01

    The forkhead box (Fox) transcription factor family has a characteristic of forkhead domain, a winged DNA-binding domain. The Fox genes have been classified into 23 subfamilies, designated FoxA to FoxS, of which the FoxR and FoxS subfamilies are specific to vertebrates. In this review, using whole-genome scanning, we identified 17 distinct Fox genes distributed on 13 chromosomes of the silkworm, Bombyx mori. A phylogenetic tree showed that the silkworm Fox genes could be classified into 13 subfamilies. The FoxK subfamily is specifically absent from the silkworm, although it is present in other lepidopteran insects, including Danaus plexippus and Heliconius melpomene. Microarray data revealed that the Fox genes have distinct expression patterns in the tissues on day 3 of the 5th instar larva. A Gene Ontology analysis suggested that the Fox genes have roles in cellular components, molecular functions, and biological processes, except in pore complex biogenesis. An analysis of the selective pressure on the proteins indicated that most of the amino acid sites in the Fox proteins are undergoing strong purifying selection. Here, we summarize the general characteristics of the Fox genes in the silkworm, which should support further functional studies of the silkworm Fox proteins.

  16. Genome-Wide Identification and Characterization of Carboxypeptidase Genes in Silkworm (Bombyx mori).

    PubMed

    Ye, Junhong; Li, Yi; Liu, Hua-Wei; Li, Jifu; Dong, Zhaoming; Xia, Qingyou; Zhao, Ping

    2016-01-01

    The silkworm (Bombyx mori) is an economically-important insect that can secrete silk. Carboxypeptidases have been found in various metazoan species and play important roles in physiological and biochemical reactions. Here, we analyzed the silkworm genome database and characterized 48 carboxypeptidases, including 34 metal carboxypeptidases (BmMCP1-BmMCP34) and 14 serine carboxypeptidases (BmSCP1-BmSCP14), to better understand their diverse functions. Compared to other insects, our results indicated that carboxypeptidases from silkworm have more family members. These silkworm carboxypeptidases could be divided into four families: Peptidase_M2 carboxypeptidases, Peptidase_M14 carboxypeptidases, Peptidase_S10 carboxypeptidases and Peptidase_S28 carboxypeptidases. Microarray analysis showed that the carboxypeptidases had distinct expression patterns, whereas quantitative real-time PCR demonstrated that the expression level of 13 carboxypeptidases significantly decreased after starvation and restored after re-feeding. Overall, our study provides new insights into the functional and evolutionary features of silkworm carboxypeptidases.

  17. The expression profile and promoter analysis of ultraspiracle gene in the silkworm Bombyx mori.

    PubMed

    Huang, Ming-xia; Du, Jie; Su, Bao-jin; Zhao, Guo-dong; Shen, Wei-de; Wei, Zheng-guo

    2014-12-01

    The nuclear receptor, ultraspiracle protein (USP), is a transcription factor and an essential component of a heterodimeric receptor complex with ecdysone receptor. However, the mechanisms underlying the transcriptional regulation of USP in silkworm are unknown. In this study, using dual-spike-in qPCR method, we examined the expression of Bombyx ultraspiracle gene (BmUSP) in various tissues of silkworm as well as expression changes after stimulation with ecdysone. The results showed that the expression levels of BmUSP gene varied in different tissues and were increased 2 h after exposure to ecdysone. To identify the molecular mechanism underlying the regulation of USP gene expression in silkworm Bombyx mori, promoter truncation analyses were performed using the luciferase reporter assay and Bac-to-Bac expression system in several tissues of B. mori. BmUSP gene promoter with 5' end serial deletions showed different levels of activity in various tissues, higher in fat body and Malpighian tubule. Deletion of the region from -485 to -445 and -307 to -281 upstream of BmUSP gene abolished and increased its promoter activity, respectively. This region contains AP-1, Dfd transcription factor binding sites. These results indicate that BmUSP are expressed at different levels in different tissues of the silkworm, but all are subjected to the regulation by ecdysone. This study would provide an important foundation for investigating the mechanism underlying the transcriptional regulation of BmUSP in the silkworm.

  18. Genome-Wide Identification and Characterization of Carboxypeptidase Genes in Silkworm (Bombyx mori)

    PubMed Central

    Ye, Junhong; Li, Yi; Liu, Hua-Wei; Li, Jifu; Dong, Zhaoming; Xia, Qingyou; Zhao, Ping

    2016-01-01

    The silkworm (Bombyx mori) is an economically-important insect that can secrete silk. Carboxypeptidases have been found in various metazoan species and play important roles in physiological and biochemical reactions. Here, we analyzed the silkworm genome database and characterized 48 carboxypeptidases, including 34 metal carboxypeptidases (BmMCP1–BmMCP34) and 14 serine carboxypeptidases (BmSCP1–BmSCP14), to better understand their diverse functions. Compared to other insects, our results indicated that carboxypeptidases from silkworm have more family members. These silkworm carboxypeptidases could be divided into four families: Peptidase_M2 carboxypeptidases, Peptidase_M14 carboxypeptidases, Peptidase_S10 carboxypeptidases and Peptidase_S28 carboxypeptidases. Microarray analysis showed that the carboxypeptidases had distinct expression patterns, whereas quantitative real-time PCR demonstrated that the expression level of 13 carboxypeptidases significantly decreased after starvation and restored after re-feeding. Overall, our study provides new insights into the functional and evolutionary features of silkworm carboxypeptidases. PMID:27483237

  19. Female qualities in males: vitellogenin synthesis induced by ovary transplants into the male silkworm, Bombyx mori.

    PubMed

    Yang, Congwen; Lin, Ying; Shen, Guanwang; Chen, Enxiang; Wang, Yanxia; Luo, Juan; Zhang, Haiyan; Xing, Runmiao; Xia, Qingyou

    2014-10-10

    Female qualities in males are common in vertebrates but have not been extensively reported in insects. Vitellogenin (Vg) is highly expressed in the female fat body and is generally required for the formation of yolk proteins in the insect egg. Vg upregulation is generally regarded as a female quality in female oviparous animals. In this study, we found that Bombyx mori Vg (BmVg) is especially highly expressed in the female pupa. Downregulation of the BmVg gene in the female pupa by RNA interference (RNAi) interfered with egg formation and embryonic development, showing the importance of BmVg in these processes. So, we used BmVg as a biomarker for female qualities in the silkworm. Hematoxylin-eosin staining and immunofluorescence histochemistry showed that ovary transplants induced BmVg synthesis in the male pupa fat body. Ovaries transplanted into male silkworms produced only a few eggs with deformed yolk granules. These results suggested that the amount of BmVg in the male silkworm was insufficient for eggs to undergo complete embryonic development. After 17-beta-estradiol was used to treat male pupae and male pupal fat bodies, BmVg was upregulated in vivo and in vitro. These findings indicated that the male silkworm has innate female qualities that were induced by a transplanted ovary and 17β-estradiol. However, in silkworms, female qualities in males are not as complete as in females.

  20. Identification and Evolution of the Silkworm Helitrons and their Contribution to Transcripts

    PubMed Central

    Han, Min-Jin; Shen, Yi-Hong; Xu, Meng-Shu; Liang, Hong-Yu; Zhang, Hua-Hao; Zhang, Ze

    2013-01-01

    In this study, we developed a structure-based approach to identify Helitrons in four lepidopterans and systematically analysed Helitrons in the silkworm genome. We found that the content of Helitrons varied greatly among genomes. The silkworm genome harboured 67 555 Helitron-related sequences that could be classified into 21 families and accounted for ∼4.23% of the genome. Thirteen of the families were new. Three families were putatively autonomous and included the replication initiator motif and helicase domain. The silkworm Helitrons were widely and randomly distributed in the genome. Most Helitron families radiated within the past 2 million years and experienced a single burst of expansion. These Helitron families captured 3724 gene fragments and contributed to at least 1.4% of the silkworm full-length cDNAs, suggesting important roles of Helitrons in the evolution of the silkworm genes. In addition, we found that some new Helitrons were generated by combinations of other Helitrons. Overall, the results presented in this study provided insights into the generation and evolution of Helitron transposons and their contribution to transcripts. PMID:23771679

  1. Genome-Wide Identification and Characterization of Carboxypeptidase Genes in Silkworm (Bombyx mori).

    PubMed

    Ye, Junhong; Li, Yi; Liu, Hua-Wei; Li, Jifu; Dong, Zhaoming; Xia, Qingyou; Zhao, Ping

    2016-01-01

    The silkworm (Bombyx mori) is an economically-important insect that can secrete silk. Carboxypeptidases have been found in various metazoan species and play important roles in physiological and biochemical reactions. Here, we analyzed the silkworm genome database and characterized 48 carboxypeptidases, including 34 metal carboxypeptidases (BmMCP1-BmMCP34) and 14 serine carboxypeptidases (BmSCP1-BmSCP14), to better understand their diverse functions. Compared to other insects, our results indicated that carboxypeptidases from silkworm have more family members. These silkworm carboxypeptidases could be divided into four families: Peptidase_M2 carboxypeptidases, Peptidase_M14 carboxypeptidases, Peptidase_S10 carboxypeptidases and Peptidase_S28 carboxypeptidases. Microarray analysis showed that the carboxypeptidases had distinct expression patterns, whereas quantitative real-time PCR demonstrated that the expression level of 13 carboxypeptidases significantly decreased after starvation and restored after re-feeding. Overall, our study provides new insights into the functional and evolutionary features of silkworm carboxypeptidases. PMID:27483237

  2. Construction of transformed, cultured silkworm cells and transgenic silkworm using the site-specific integrase system from phage φC31.

    PubMed

    Yin, Yajuan; Cao, Guangli; Xue, Renyu; Gong, Chengliang

    2014-10-01

    The Streptomyces bacteriophage, φC31, uses a site-specific integrase enzyme to perform efficient recombination. The recombination system uses specific sequences to integrate exogenous DNA from the phage into a host. The sequences are known as the attP site in the phage and the attB site in the host. The system can be used as a genetic manipulation tool. In this study it has been applied to the transformation of cultured BmN cells and the construction of transgenic Bombyx mori individuals. A plasmid, pSK-attB/Pie1-EGFP/Zeo-PASV40, containing a cassette designed to express a egfp-zeocin fusion gene, was co-transfected into cultured BmN cells with a helper plasmid, pSK-Pie1/NLS-Int/NSL. Expression of the egfp-zeocin fusion gene was driven by an ie-1 promoter, downstream of a φC31 attB site. The helper plasmid encoded the φC31 integrase enzyme, which was flanked by two nuclear localization signals. Expression of the egfp-zeocin fusion gene could be observed in transformed cells. The two plasmids were also transferred into silkworm eggs to obtain transgenic silkworms. Successful integration of the fusion gene was indicated by the detection of green fluorescence, which was emitted by the silkworms. Nucleotide sequence analysis demonstrated that the attB site had been cut, to allow recombination between the attB and endogenous pseudo attP sites in the cultured silkworm cells and silkworm individuals.

  3. Efficient large-scale protein production of larvae and pupae of silkworm by Bombyx mori nuclear polyhedrosis virus bacmid system.

    PubMed

    Motohashi, Tomoko; Shimojima, Tsukasa; Fukagawa, Tatsuo; Maenaka, Katsumi; Park, Enoch Y

    2005-01-21

    Silkworm is one of the most attractive hosts for large-scale production of eukaryotic proteins as well as recombinant baculoviruses for gene transfer to mammalian cells. The bacmid system of Autographa californica nuclear polyhedrosis virus (AcNPV) has already been established and widely used. However, the AcNPV does not have a potential to infect silkworm. We developed the first practical Bombyx mori nuclear polyhedrosis virus bacmid system directly applicable for the protein expression of silkworm. By using this system, the green fluorescence protein was successfully expressed in silkworm larvae and pupae not only by infection of its recombinant virus but also by direct injection of its bacmid DNA. This method provides the rapid protein production in silkworm as long as 10 days, is free from biohazard, thus will be a powerful tool for the future production factory of recombinant eukaryotic proteins and baculoviruses. PMID:15596136

  4. Learning experiences at Oak Ridge

    SciTech Connect

    White, R.K.

    1990-01-01

    The Oak Ridge Operations (ORO) of DOE has organized an Environmental Restoration Program to handle environmental cleanup activitis for the Oak Ridge Reservation (ORR) following General Watkins' reorganization at DOE Headquarters. Based on the major facilities and locations of contamination sites, the Environmental Restoration Program is divided into five subprograms: Oak Ridge, National Laboatory (ORNL) sites, y-12 Plant sites, Oak Ridge Gaseous Diffusion Plant (ORGDP) sites, Oak Ridge Associated Universities (ORAU) sites and off-site areas. The Office of Risk Analysis at ORNL was established under the auspices of the Environmental Restoration Program to implement Superfun legislation in the five subprograms of DOE-ORO. Risk assessment must examine protetial human health and ecological impacts from contaminant sources that range from highly radioactive materials to toxic chemicals and mixed wastes. The remedial alternatives we are evaluating need to reach acceptable levels of risk effectively while also being cost-efficient. The purpose of this paper is to highlight areas of particular interest and concern at Oak Ridge and to discuss, where possible, solutions implemented by the Oak Ridge Environmental Restoation Program.

  5. Effect of Venom from the Jellyfish Nemopilema nomurai on the Silkworm Bombyx mori L.

    PubMed

    Yu, Huahua; Li, Rongfeng; Chen, Xiaolin; Yue, Yang; Xing, Ronge; Liu, Song; Li, Pengcheng

    2015-10-01

    The silkworm Bombyx mori L. (B. mori) has a significant impact on the economy by producing more than 80% of the globally produced raw silk. The exposure of silkworm to pesticides may cause adverse effects on B. mori, such as a reduction in the production and quality of silk. This study aims to assay the effect of venom from the jellyfish Nemopilema nomurai on growth, cuticle and acetylcholinesterase (AChE) activity of the silkworm B. mori by the leaf dipping method. The experimental results revealed that the four samples caused neither antifeeding nor a lethal effect on B. mori. The sample SFV inhibited B. mori growth after 6 days of treatment in a dose-dependent manner. The samples SFV, DSFV and Fr-1 inhibited the precipitation and synthesis of chitin in the cuticle after 12 and 14 days of treatment. In the case of the four samples, the AChE was significantly improved after 14 days of treatment.

  6. The extramacrochaetae gene is required for blastokinesis in silkworm, Bombyx mori.

    PubMed

    Liu, Wenbin; Chai, Dezhi; Wang, Cailian; Li, Qing; Lei, Jinfeng; Yang, Min; Dai, Fangyin; Lu, Cheng

    2015-07-01

    In silkworm, Bombyx mori Linnaeus (Lepidoptera: Bombycidae), blastokinesis results in embryo reversal from ventrally to dorsally convex flexion. In this study, we showed that the extramacrochaetae (emc) gene is required for blastokinesis in silkworm. Depletion of Bmemc expression via RNA interference led to severe phenotypic defects in blastokinesis. The defective embryos failed to invert their body sides during blastokinesis. This caused the posterior half of the abdomen to abnormally fold back toward the dorsal side, forming a U-shaped morphology. Dorsal closure was also disrupted. Our results suggest that Bmemc is involved in blastokinesis of silkworm embryos. J. Exp. Zool. (Mol. Dev. Evol.) 324B: 405-409, 2015. © 2015 Wiley Periodicals, Inc.

  7. Effect of Venom from the Jellyfish Nemopilema nomurai on the Silkworm Bombyx mori L

    PubMed Central

    Yu, Huahua; Li, Rongfeng; Chen, Xiaolin; Yue, Yang; Xing, Ronge; Liu, Song; Li, Pengcheng

    2015-01-01

    The silkworm Bombyx mori L. (B. mori) has a significant impact on the economy by producing more than 80% of the globally produced raw silk. The exposure of silkworm to pesticides may cause adverse effects on B. mori, such as a reduction in the production and quality of silk. This study aims to assay the effect of venom from the jellyfish Nemopilema nomurai on growth, cuticle and acetylcholinesterase (AChE) activity of the silkworm B. mori by the leaf dipping method. The experimental results revealed that the four samples caused neither antifeeding nor a lethal effect on B. mori. The sample SFV inhibited B. mori growth after 6 days of treatment in a dose-dependent manner. The samples SFV, DSFV and Fr-1 inhibited the precipitation and synthesis of chitin in the cuticle after 12 and 14 days of treatment. In the case of the four samples, the AChE was significantly improved after 14 days of treatment. PMID:26404374

  8. Effect of Venom from the Jellyfish Nemopilema nomurai on the Silkworm Bombyx mori L.

    PubMed

    Yu, Huahua; Li, Rongfeng; Chen, Xiaolin; Yue, Yang; Xing, Ronge; Liu, Song; Li, Pengcheng

    2015-10-01

    The silkworm Bombyx mori L. (B. mori) has a significant impact on the economy by producing more than 80% of the globally produced raw silk. The exposure of silkworm to pesticides may cause adverse effects on B. mori, such as a reduction in the production and quality of silk. This study aims to assay the effect of venom from the jellyfish Nemopilema nomurai on growth, cuticle and acetylcholinesterase (AChE) activity of the silkworm B. mori by the leaf dipping method. The experimental results revealed that the four samples caused neither antifeeding nor a lethal effect on B. mori. The sample SFV inhibited B. mori growth after 6 days of treatment in a dose-dependent manner. The samples SFV, DSFV and Fr-1 inhibited the precipitation and synthesis of chitin in the cuticle after 12 and 14 days of treatment. In the case of the four samples, the AChE was significantly improved after 14 days of treatment. PMID:26404374

  9. Silkworms culture as a source of protein for humans in space

    NASA Astrophysics Data System (ADS)

    Yang, Yunan; Tang, Liman; Tong, Ling; Liu, Hong

    2009-04-01

    This paper focuses on the problem about a configuration with complete nutrition for humans in a Controlled Ecological Life Support System (CELSS) applied in the spacebases. The possibility of feeding silkworms to provide edible animal protein with high quality for taikonauts during long-term spaceflights and lunar-based missions was investigated from several aspects, including the nutrition structure of silkworms, feeding method, processing methods, feeding equipment, growing conditions and the influences on the space environmental condition changes caused by the silkworms. The originally inedible silk is also regarded as a protein source. A possible process of edible silk protein was brought forward in this paper. After being processed, the silk can be converted to edible protein for humans. The conclusion provides a promising approach to solving the protein supply problem for the taikonauts living in space during an extended exploration period.

  10. The advances and perspectives of recombinant protein production in the silk gland of silkworm Bombyx mori.

    PubMed

    Xu, Hanfu

    2014-10-01

    The silk gland of silkworm Bombyx mori, is one of the most important organs that has been fully studied and utilized so far. It contributes finest silk fibers to humankind. The silk gland has excellent ability of synthesizing silk proteins and is a kind tool to produce some useful recombinant proteins, which can be widely used in the biological, biotechnical and pharmaceutical application fields. It's a very active area to express recombinant proteins using the silk gland as a bioreactor, and great progress has been achieved recently. This review recapitulates the progress of producing recombinant proteins and silk-based biomaterials in the silk gland of silkworm in addition to the construction of expression systems. Current challenges and future trends in the production of valuable recombinant proteins using transgenic silkworms are also discussed.

  11. Production and characterization of the celery mismatch endonuclease CEL II using baculovirus/silkworm expression system.

    PubMed

    Mon, Hiroaki; Lee, Jaeman; Fukushima, Mai; Nagata, Yudai; Fujii, Mie; Xu, Jian; Nishi, Oumi; Iiyama, Kazuhiro; Kusakabe, Takahiro

    2013-08-01

    Mutation and polymorphism detection by nucleases has become a more important tool in clinical and biological researches. There are several kinds of single-stranded nucleases for detecting mismatched DNAs. One of them, CEL II, was isolated from Apium graveolens and cleaves DNA with high specificity at sites of mismatch. High-throughput mutation scanning requires large quantity of CEL II endonuclease. Here, we demonstrate high-level expression of CEL II using silkworm-baculovirus system. The recombinant CEL II secreted in silkworm hemolymph was glycosylated and susceptible to N-glycosidase F. Additionally, larger metal ions such as Ca(2+) and Sr(2+) were able to replace Mg(2+) and enhanced mismatch cleavage activity of CEL II. These results indicate that the silkworm-baculovirus platform is a good alternative system to obtain the functional CEL II.

  12. Two Hemocyte Lineages Exist in Silkworm Larval Hematopoietic Organ

    PubMed Central

    Nakahara, Yuichi; Kanamori, Yasushi; Kiuchi, Makoto; Kamimura, Manabu

    2010-01-01

    Background Insects have multiple hemocyte morphotypes with different functions as do vertebrates, however, their hematopoietic lineages are largely unexplored with the exception of Drosophila melanogaster. Methodology/Principal Findings To study the hematopoietic lineage of the silkworm, Bombyx mori, we investigated in vivo and in vitro differentiation of hemocyte precursors in the hematopoietic organ (HPO) into the four mature hemocyte subsets, namely, plasmatocytes, granulocytes, oenocytoids, and spherulocytes. Five days after implantation of enzymatically-dispersed HPO cells from a GFP-expressing transgenic line into the hemocoel of normal larvae, differentiation into plasmatocytes, granulocytes and oenocytoids, but not spherulocytes, was observed. When the HPO cells were cultured in vitro, plasmatocytes appeared rapidly, and oenocytoids possessing prophenol oxidase activity appeared several days later. HPO cells were also able to differentiate into a small number of granulocytes, but not into spherulocytes. When functionally mature plasmatocytes were cultured in vitro, oenocytoids were observed 10 days later. These results suggest that the hemocyte precursors in HPO first differentiate into plasmatocytes, which further change into oenocytoids. Conclusions/Significance From these results, we propose that B. mori hemocytes can be divided into two major lineages, a granulocyte lineage and a plasmatocyte-oenocytoid lineage. The origins of the spherulocytes could not be determined in this study. We construct a model for the hematopoietic lineages at the larval stage of B. mori. PMID:20676370

  13. Molecular mapping and characterization of the silkworm apodal mutant

    PubMed Central

    Chen, Peng; Tong, Xiao-Ling; Fu, Ming-Yue; Hu, Hai; Song, Jiang-Bo; He, Song-Zhen; Gai, Ting-Ting; Dai, Fang-Yin; Lu, Cheng

    2016-01-01

    The morphological diversity of insects is important for their survival; in essence, it results from the differential expression of genes during development of the insect body. The silkworm apodal (ap) mutant has degraded thoracic legs making crawling and eating difficult and the female is sterile, which is an ideal subject for studying the molecular mechanisms of morphogenesis. Here, we confirmed that the infertility of ap female moths is a result of the degradation of the bursa copulatrix. Positional cloning of ap locus and expression analyses reveal that the Bombyx mori sister of odd and bowl (Bmsob) gene is a strong candidate for the ap mutant. The expression of Bmsob is down-regulated, while the corresponding Hox genes are up-regulated in the ap mutant compared to the wild type. Analyses with the dual luciferase assay present a declined activity of the Bmsob promoter in the ap mutant. Furthermore, we demonstrate that Bmsob can inhibit Hox gene expression directly and by suppressing the expression of other genes, including the BmDsp gene. The results of this study are an important contribution to our understanding of the diversification of insect body plan. PMID:26738847

  14. Thermally induced increase in energy transport capacity of silkworm silks.

    PubMed

    Liu, Guoqing; Xu, Shen; Cao, Ting-Ting; Lin, Huan; Tang, Xiaoduan; Zhang, Yu-Qing; Wang, Xinwei

    2014-10-01

    This work reports on the first study of thermally induced effect on energy transport in single filaments of silkworm (Bombyx mori) fibroin degummed mild (type 1), moderate (type 2), to strong (type 3). After heat treatment from 140 to 220°C, the thermal diffusivity of silk fibroin type 1, 2, and 3 increases up to 37.9, 20.9, and 21.5%, respectively. Our detailed scanning electron microscopy study confirms that the sample diameter change is almost negligible before and after heat treatment. Raman analysis is performed on the original and heat-treated (at 147°C) samples. After heat treatment at 147°C, the Raman peaks at 1081, 1230, and 1665 cm(-1) become stronger and narrower, indicating structural transformation from amorphous to crystalline. A structure model composed of amorphous, crystalline, and laterally ordered regions is proposed to explain the structural change by heat treatment. Owing to the close packing of more adjacent laterally ordered regions, the number and size of the crystalline regions of Bombyx mori silk fibroin increase by heat treatment. This structure change gives the observed significant thermal diffusivity increase by heat treatment.

  15. The effects of diphtheria toxin on the Cecropia silkworm.

    PubMed

    PAPPENHEIMER, A M; WILLIAMS, C M

    1952-05-01

    1. The metamorphosis of the Cecropia silkworm is accompanied by large and systematic changes in the insect's sensitivity to diphtheria toxin. 2. Injection of less than 1 gamma of toxin into mature caterpillars, prepupae, or developing adults causes cessation of development followed by delayed death 1 to 5 weeks later. 3. Dormant pupae, on the contrary, are resistant to 70 gamma of toxin and may survive even this enormous dose for over 4 weeks. One-hundredth of this dose, however, prevents pupae from initiating adult development. 4. Tetanus toxin, to which the insect is insensitive, failed to duplicate any of these effects. 5. Maximal sensitivity to diphtheria toxin is characteristic of those stages in the life history which depend on the presence and function of the cytochrome system. Resistance to the toxin, as in the case of the diapausing pupa, is correlated with the existence and utilization of metabolic pathways other than the usual cytochrome system. 6. This correlation persists within the individual insect. Thus, within the diapausing pupa, the toxin fails to affect the heart in which a normal cytochrome system is absent, but, within the same insect, causes a degeneration of the intersegmental muscles in which an intact cytochrome system is present. 7. These several lines of evidence are interpreted in support of the conclusion that diphtheria toxin acts by blocking the synthesis of one or more components in the cytochrome system. PMID:14955616

  16. Molecular mapping and characterization of the silkworm apodal mutant.

    PubMed

    Chen, Peng; Tong, Xiao-Ling; Fu, Ming-Yue; Hu, Hai; Song, Jiang-Bo; He, Song-Zhen; Gai, Ting-Ting; Dai, Fang-Yin; Lu, Cheng

    2016-01-01

    The morphological diversity of insects is important for their survival; in essence, it results from the differential expression of genes during development of the insect body. The silkworm apodal (ap) mutant has degraded thoracic legs making crawling and eating difficult and the female is sterile, which is an ideal subject for studying the molecular mechanisms of morphogenesis. Here, we confirmed that the infertility of ap female moths is a result of the degradation of the bursa copulatrix. Positional cloning of ap locus and expression analyses reveal that the Bombyx mori sister of odd and bowl (Bmsob) gene is a strong candidate for the ap mutant. The expression of Bmsob is down-regulated, while the corresponding Hox genes are up-regulated in the ap mutant compared to the wild type. Analyses with the dual luciferase assay present a declined activity of the Bmsob promoter in the ap mutant. Furthermore, we demonstrate that Bmsob can inhibit Hox gene expression directly and by suppressing the expression of other genes, including the BmDsp gene. The results of this study are an important contribution to our understanding of the diversification of insect body plan. PMID:26738847

  17. Transcriptome analysis of interactions between silkworm and cytoplasmic polyhedrosis virus

    PubMed Central

    Jiang, Liang; Peng, Zhengwen; Guo, Youbing; Cheng, Tingcai; Guo, Huizhen; Sun, Qiang; Huang, Chunlin; Zhao, Ping; Xia, Qingyou

    2016-01-01

    Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) specifically infects silkworm midgut (MG) and multiplication occurs mainly in posterior midgut (PM). In this study, MG and fat body (FB) were extracted at 0, 3, 24, and 72 h after BmCPV infection. The total sequence reads of each sample were more than 1510000, and the mapping ratio exceeded 95.3%. Upregulated transcripts increased in MG during the infection process. Gene ontology (GO) categories showed that antioxidants were all upregulated in FB but not in MG. BGI001299, BGI014434, BGI012068, and BGI009201 were MG-specific genes with transmembrane transport function, the expression of which were induced by BmCPV. BGI001299, BGI014434, and BGI012068 expressed in entire MG and may be involved in BmCPV invasion. BGI009201 expressed only in PM and may be necessary for BmCPV proliferation. BmPGRP-S2 and BGI012452 (a putative serine protease) were induced by BmCPV and may be involved in immune defense against BmCPV. The expression level of BmCPV S1, S2, S3, S6, and S7 was high and there was no expression of S9 in MG 72 h, implying that the expression time of structural protein coding genes is earlier. These results provide insights into the mechanism of BmCPV infection and host defense. PMID:27118345

  18. Evaluation of anti-diabetic drugs by using silkworm, Bombyx mori.

    PubMed

    Matsumoto, Yasuhiko; Sekimizu, Kazuhisa

    2016-02-01

    Since diabetes patients are increasing in the world, establishment of a novel method for development of anti-diabetic drugs is desired. In this review, we describe recent results of our studies regarding diabetic model using silkworms for evaluation of anti-diabetic drugs for patients of both type I and type II diabetes. The use of the evaluation systems using silkworms is expected to contribute to reduction in cost and in the number of mammals sacrificed for screening of anti-diabetic drugs.

  19. [Cloning of silkworm 3-hydroxyisobutyrate dehydrogenase gene and its expression patterns analysis in simulated weightless environment].

    PubMed

    Tian, Zongcheng; Zhou, Bo; Qian, Airong; Xu, Huiyun; Di, Shengmeng; Zhao, Yunpo; Zhang, Yuping; Liu, Jia; Huang, Yongping; Shang, Peng

    2008-12-01

    The full length cDNA of silkworm hibadh gene was cloned by RT-PCR and RACE (Rapid amplification of cDNA ends) technique. The hibadh gene and its deduced amino acid sequences were analyzed. The tissue distribution of hibadh gene in 5th instar silkworm larvae was tested by RT-PCR. The expression patterns of hibadh gene in simulated weightless environment were analyzed by real time RT-PCR. The results showed that the full length hibadh cDNA sequence was 1074 bp in lenth, including an open read frame of 969 bp encoding the entire coding region of Hibadh (GenBank accession No. EU719652). The deduced amino acid sequence similarities of hibadh between silkworm and Burkholderia ambifaria, Drosophila melanogaster, Apis mellifera, Xenopus tropicalis, Mus musculus, Homo sapiens were 46%, 43%, 48%, 44%, 45%, 45%, respectively. Signal peptide analysis showed that Hibadh was a secretory protein. There wasn't glycosyl-phosphatidyl inositol anchor site in Hibadh amino acid sequence. Molecular weight and isoelectric point of Hibadh were 34.1 kD and 9.14 respectively. The RT-PCR tests indicated that the hibadh gene expressed in head, silk gland, midgut, cuticle, blood, fat body, tuba malpighii of the 5th instar silkworm larvae. There were different expression patterns of hibadh gene during different silkworm embryo period in simulated weightless environment. Simulated weightlessness resulted in the expression of silkworm hibadh gene up regulated 2.3-fold (P < 0.05), up regulated 4.6-fold (P<0.01), down regulated 7.6-fold (P < 0.01), down regulated 2.6-fold (P < 0.05) during apophysis formation period, inverse period, trachea formation period, and whole embryo period, respectively. There was no significant change of hibadh gene expression during other period of silkworm embryo between simulated weightless and control groups. There were different response patterns to simulated weightless environment between hibadh gene and whole body of silkworm. Gene showed much higher sensitivity

  20. Poison Ivy, Oak, and Sumac

    MedlinePlus

    ... U.S.) is a delayed allergic reaction. Brushing the plant on the skin results in blisters and slightly ... of poison ivy, poison oak, and poison sumac plants. People typically have itchy bumps (papules) and blisters ( ...

  1. Contact sensitivity to oak moss.

    PubMed

    Gonçalo, S; Cabral, F; Gonçalo, M

    1988-11-01

    Oak moss allergy was the principle allergen in contact sensitivity to perfumes (45%); 31 patients reacting to oak moss were studied. The sensitivity was attributed to contact with perfumes in 20, lichens in 7 and unknown in 4. Atranorin was the most frequent allergen, followed by usnic, evernic and fumarprotocetraric acids. Concomitant allergy occurred to several lichen acids and also to balsam of Peru, colophony and other fragrance components.

  2. Contact sensitivity to oak moss.

    PubMed

    Gonçalo, S; Cabral, F; Gonçalo, M

    1988-11-01

    Oak moss allergy was the principle allergen in contact sensitivity to perfumes (45%); 31 patients reacting to oak moss were studied. The sensitivity was attributed to contact with perfumes in 20, lichens in 7 and unknown in 4. Atranorin was the most frequent allergen, followed by usnic, evernic and fumarprotocetraric acids. Concomitant allergy occurred to several lichen acids and also to balsam of Peru, colophony and other fragrance components. PMID:3233955

  3. Allergenic Characterization of 27-kDa Glycoprotein, a Novel Heat Stable Allergen, from the Pupa of Silkworm, Bombyx mori

    PubMed Central

    Son, Mina; Lee, June Yong

    2016-01-01

    Boiled silkworm pupa is a traditional food in Asia, and patients with silkworm pupa food allergy are common in these regions. Still now only one allergen from silkworm, arginine kinase, has been identified. The purpose of this study was to identify novel food allergens in silkworm pupa by analyzing a protein extract after heat treatment. Heat treated extracts were examined by proteomic analysis. A 27-kDa glycoprotein was identified, expressed in Escherichia coli, and purified. IgE reactivity of the recombinant protein was investigated by ELISA. High molecular weight proteins (above 100 kDa) elicited increased IgE binding after heat treatment compared to that before heat treatment. The molecular identities of these proteins, however, could not be determined. IgE reactivity toward a 27-kDa glycoprotein was also increased after heating the protein extract. The recombinant protein was recognized by IgE antibodies from allergic subjects (33.3%). Glycation or aggregation of protein by heating may create new IgE binding epitopes. Heat stable allergens are shown to be important in silkworm allergy. Sensitization to the 27-kDa glycoprotein from silkworm may contribute to elevation of IgE to silkworm. PMID:26770033

  4. Postoperative anti-adhesion ability of a novel carboxymethyl chitosan from silkworm pupa in a rat cecal abrasion model.

    PubMed

    Zhu, Lin; Zhang, Yu-Qing

    2016-04-01

    N,O-Carboxymethyl chitosan (NOCC) can prevent postsurgical adhesion formation. Here, we described the preparation of a novel silkworm pupa NOCC and its effects on the prevention of postoperative adhesion in a rat cecal abrasion model. The degree of deacetylation (DDA) of silkworm pupa chitosan was only 49.87 ± 0.86%; regardless, it was used as the raw material to construct the novel silkworm pupa NOCC, which had a weaker crystallinity than the NOCC standard. Sixty male Sprague-Dawley rats were divided into three groups and treated as follows: 0.9% normal saline solution as a negative control, medical anti-adhesion gel as a positive control and the silkworm pupa NOCC anti-adhesion solution. Two and three weeks after surgery, the animals were killed and the adhesion formation was scored. The silkworm pupa NOCC solution significantly decreased the levels of WBC, TNF-α, IL-1β, IL-2, IL-6 and IL-8 but had no effect on IL-4. Additionally, a lower level of TGF-β1 expression was found in the silkworm pupa NOCC group, and significantly less collagen (P<0.01) and fewer inflammatory cells and fibroblasts were detected in the animals of this group. These results suggested that the novel NOCC from silkworm pupa using the method described here have potential applications in the prevention of postoperative intestinal adhesion. PMID:26838865

  5. Allergenic Characterization of 27-kDa Glycoprotein, a Novel Heat Stable Allergen, from the Pupa of Silkworm, Bombyx mori.

    PubMed

    Jeong, Kyoung Yong; Son, Mina; Lee, June Yong; Park, Kyung Hee; Lee, Jae-Hyun; Park, Jung-Won

    2016-01-01

    Boiled silkworm pupa is a traditional food in Asia, and patients with silkworm pupa food allergy are common in these regions. Still now only one allergen from silkworm, arginine kinase, has been identified. The purpose of this study was to identify novel food allergens in silkworm pupa by analyzing a protein extract after heat treatment. Heat treated extracts were examined by proteomic analysis. A 27-kDa glycoprotein was identified, expressed in Escherichia coli, and purified. IgE reactivity of the recombinant protein was investigated by ELISA. High molecular weight proteins (above 100 kDa) elicited increased IgE binding after heat treatment compared to that before heat treatment. The molecular identities of these proteins, however, could not be determined. IgE reactivity toward a 27-kDa glycoprotein was also increased after heating the protein extract. The recombinant protein was recognized by IgE antibodies from allergic subjects (33.3%). Glycation or aggregation of protein by heating may create new IgE binding epitopes. Heat stable allergens are shown to be important in silkworm allergy. Sensitization to the 27-kDa glycoprotein from silkworm may contribute to elevation of IgE to silkworm.

  6. Postoperative anti-adhesion ability of a novel carboxymethyl chitosan from silkworm pupa in a rat cecal abrasion model.

    PubMed

    Zhu, Lin; Zhang, Yu-Qing

    2016-04-01

    N,O-Carboxymethyl chitosan (NOCC) can prevent postsurgical adhesion formation. Here, we described the preparation of a novel silkworm pupa NOCC and its effects on the prevention of postoperative adhesion in a rat cecal abrasion model. The degree of deacetylation (DDA) of silkworm pupa chitosan was only 49.87 ± 0.86%; regardless, it was used as the raw material to construct the novel silkworm pupa NOCC, which had a weaker crystallinity than the NOCC standard. Sixty male Sprague-Dawley rats were divided into three groups and treated as follows: 0.9% normal saline solution as a negative control, medical anti-adhesion gel as a positive control and the silkworm pupa NOCC anti-adhesion solution. Two and three weeks after surgery, the animals were killed and the adhesion formation was scored. The silkworm pupa NOCC solution significantly decreased the levels of WBC, TNF-α, IL-1β, IL-2, IL-6 and IL-8 but had no effect on IL-4. Additionally, a lower level of TGF-β1 expression was found in the silkworm pupa NOCC group, and significantly less collagen (P<0.01) and fewer inflammatory cells and fibroblasts were detected in the animals of this group. These results suggested that the novel NOCC from silkworm pupa using the method described here have potential applications in the prevention of postoperative intestinal adhesion.

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

    PubMed

    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.

  8. Allergenic Characterization of 27-kDa Glycoprotein, a Novel Heat Stable Allergen, from the Pupa of Silkworm, Bombyx mori.

    PubMed

    Jeong, Kyoung Yong; Son, Mina; Lee, June Yong; Park, Kyung Hee; Lee, Jae-Hyun; Park, Jung-Won

    2016-01-01

    Boiled silkworm pupa is a traditional food in Asia, and patients with silkworm pupa food allergy are common in these regions. Still now only one allergen from silkworm, arginine kinase, has been identified. The purpose of this study was to identify novel food allergens in silkworm pupa by analyzing a protein extract after heat treatment. Heat treated extracts were examined by proteomic analysis. A 27-kDa glycoprotein was identified, expressed in Escherichia coli, and purified. IgE reactivity of the recombinant protein was investigated by ELISA. High molecular weight proteins (above 100 kDa) elicited increased IgE binding after heat treatment compared to that before heat treatment. The molecular identities of these proteins, however, could not be determined. IgE reactivity toward a 27-kDa glycoprotein was also increased after heating the protein extract. The recombinant protein was recognized by IgE antibodies from allergic subjects (33.3%). Glycation or aggregation of protein by heating may create new IgE binding epitopes. Heat stable allergens are shown to be important in silkworm allergy. Sensitization to the 27-kDa glycoprotein from silkworm may contribute to elevation of IgE to silkworm. PMID:26770033

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

    PubMed

    Zhao, Hailong; Zheng, Chunqin; Cui, Hongjuan

    2015-01-01

    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(var)3-9 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.

  10. Hedgehog signaling pathway regulated the target genes for adipogenesis in silkworm Bombyx mori.

    PubMed

    Liang, Shuang; Chen, Rui-Ting; Zhang, Deng-Pan; Xin, Hu-Hu; Lu, Yan; Wang, Mei-Xian; Miao, Yun-Gen

    2015-10-01

    Hedgehog (Hh) signals regulate invertebrate and vertebrate development, yet the role of the pathway in adipose development remains poorly understood. In this report, we found that Hh pathway components are expressed in the fat body of silkworm larvae. Functional analysis of these components in a BmN cell line model revealed that activation of the Hh gene stimulated transcription of Hh pathway components, but inhibited the expression of the adipose marker gene AP2. Conversely, specific RNA interference-mediated knockdown of Hh resulted in increased AP2 expression. This further showed the regulation of Hh signal on the adipose marker gene. In silkworm larval models, enhanced adipocyte differentiation and an increase in adipocyte cell size were observed in silkworms that had been treated with a specific Hh signaling pathway antagonist, cyclopamine. The fat-body-specific Hh blockade tests were consistent with Hh signaling inhibiting silkworm adipogenesis. Our results indicate that the role of Hh signaling in inhibiting fat formation is conserved in vertebrates and invertebrates.

  11. High-Toughness Silk Produced by a Transgenic Silkworm Expressing Spider (Araneus ventricosus) Dragline Silk Protein

    PubMed Central

    Kuwana, Yoshihiko; Sezutsu, Hideki; Nakajima, Ken-ichi; Tamada, Yasushi; Kojima, Katsura

    2014-01-01

    Spider dragline silk is a natural fiber that has excellent tensile properties; however, it is difficult to produce artificially as a long, strong fiber. Here, the spider (Araneus ventricosus) dragline protein gene was cloned and a transgenic silkworm was generated, that expressed the fusion protein of the fibroin heavy chain and spider dragline protein in cocoon silk. The spider silk protein content ranged from 0.37 to 0.61% w/w (1.4–2.4 mol%) native silkworm fibroin. Using a good silk-producing strain, C515, as the transgenic silkworm can make the raw silk from its cocoons for the first time. The tensile characteristics (toughness) of the raw silk improved by 53% after the introduction of spider dragline silk protein; the improvement depended on the quantity of the expressed spider dragline protein. To demonstrate the commercial feasibility for machine reeling, weaving, and sewing, we used the transgenic spider silk to weave a vest and scarf; this was the first application of spider silk fibers from transgenic silkworms. PMID:25162624

  12. Intravital imaging of Bacillus thuringiensis Cry1A toxin binding sites in the midgut of silkworm.

    PubMed

    Li, Na; Wang, Jing; Han, Heyou; Huang, Liang; Shao, Feng; Li, Xuepu

    2014-02-15

    Identification of the resistance mechanism of insects against Bacillus thuringiensis Cry1A toxin is becoming an increasingly challenging task. This fact highlights the need for establishing new methods to further explore the molecular interactions of Cry1A toxin with insects and the receptor-binding region of Cry1A toxins for their wider application as biopesticides and a gene source for gene-modified crops. In this contribution, a quantum dot-based near-infrared fluorescence imaging method has been applied for direct dynamic tracking of the specific binding of Cry1A toxins, CrylAa and CrylAc, to the midgut tissue of silkworm. The in vitro fluorescence imaging displayed the higher binding specificity of CrylAa-QD probes compared to CrylAc-QD to the brush border membrane vesicles of midgut from silkworm. The in vivo imaging demonstrated that more CrylAa-QDs binding to silkworm midgut could be effectively and distinctly monitored in living silkworms. Furthermore, frozen section analysis clearly indicated the broader receptor-binding region of Cry1Aa compared to that of Cry1Ac in the midgut part. These observations suggest that the insecticidal activity of Cry toxins may depend on the receptor-binding sites, and this scatheless and visual near-infrared fluorescence imaging could provide a new avenue to study the resistance mechanism to maintain the insecticidal activity of B. thuringiensis toxins. PMID:24252542

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

    PubMed Central

    Zhao, Hailong; Zheng, Chunqin; Cui, Hongjuan

    2015-01-01

    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(var)3-9 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. PMID:26558257

  14. High-toughness silk produced by a transgenic silkworm expressing spider (Araneus ventricosus) dragline silk protein.

    PubMed

    Kuwana, Yoshihiko; Sezutsu, Hideki; Nakajima, Ken-ichi; Tamada, Yasushi; Kojima, Katsura

    2014-01-01

    Spider dragline silk is a natural fiber that has excellent tensile properties; however, it is difficult to produce artificially as a long, strong fiber. Here, the spider (Araneus ventricosus) dragline protein gene was cloned and a transgenic silkworm was generated, that expressed the fusion protein of the fibroin heavy chain and spider dragline protein in cocoon silk. The spider silk protein content ranged from 0.37 to 0.61% w/w (1.4-2.4 mol%) native silkworm fibroin. Using a good silk-producing strain, C515, as the transgenic silkworm can make the raw silk from its cocoons for the first time. The tensile characteristics (toughness) of the raw silk improved by 53% after the introduction of spider dragline silk protein; the improvement depended on the quantity of the expressed spider dragline protein. To demonstrate the commercial feasibility for machine reeling, weaving, and sewing, we used the transgenic spider silk to weave a vest and scarf; this was the first application of spider silk fibers from transgenic silkworms.

  15. Large-scale production of bioactive recombinant human acidic fibroblast growth factor in transgenic silkworm cocoons

    PubMed Central

    Wang, Feng; Wang, Riyuan; Wang, Yuancheng; Zhao, Ping; Xia, Qingyou

    2015-01-01

    With an increasing clinical demand for functional therapeutic proteins every year, there is an increasing requirement for the massive production of bioactive recombinant human acidic fibroblast growth factor (r-haFGF). In this present study, we delicately explore a strategy for the mass production of r-haFGF protein with biological activity in the transgenic silkworm cocoons. The sequence-optimized haFGF was inserted into an enhanced sericin-1 expression system to generate the original transgenic silkworm strain, which was then further crossed with a PIG jumpstarter strain to achieve the remobilization of the expression cassette to a “safe harbor” locus in the genome for the efficient expression of r-haFGF. In consequence, the expression of r-haFGF protein in the mutant line achieved a 5.6-fold increase compared to the original strain. The high content of r-haFGF facilitated its purification and large-scald yields. Furthermore, the r-haFGF protein bioactively promoted the growth, proliferation and migration of NIH/3T3 cells, suggesting the r-haFGF protein possessed native mitogenic activity and the potential for wound healing. These results show that the silk gland of silkworm could be an efficient bioreactor strategy for recombinant production of bioactive haFGF in silkworm cocoons. PMID:26567460

  16. Identification of a Serratia marcescens virulence factor that promotes hemolymph bleeding in the silkworm, Bombyx mori.

    PubMed

    Ishii, Kenichi; Adachi, Tatsuo; Hara, Takashi; Hamamoto, Hiroshi; Sekimizu, Kazuhisa

    2014-03-01

    Injection of culture supernatant of Serratia marcescens, a Gram-negative bacterium pathogenic to a wide range of host animals including insects and mammals, into the hemolymph of silkworm (Bombyx mori) larvae led to continuous flow of the hemolymph (blood of insects) from the injection site. The amount of hemolymph lost within 60 min reached 15-20% of the total larval weight. Using a bioassay with live silkworms, we purified Serralysin, a metalloprotease that requires divalent cations for its activity, as the factor responsible for the promotion of hemolymph bleeding from the culture supernatant of S. marcescens. Recombinant protein also induced hemolymph bleeding in silkworms. Moreover, the culture supernatant of an S. marcescens disruption mutant of the ser gene showed attenuated ability to promote hemolymph bleeding. In addition, this bleeding-promoting activity of the S. marcescens culture supernatant was attenuated by disruption of the wecA gene, which is involved in the biosynthesis of the lipopolysaccharide O-antigen. These findings suggest that Serralysin metalloprotease contributes to the pathogenesis of S. marcescens by inhibiting wound healing, which leads to a massive loss of hemolymph from silkworm larvae.

  17. High-toughness silk produced by a transgenic silkworm expressing spider (Araneus ventricosus) dragline silk protein.

    PubMed

    Kuwana, Yoshihiko; Sezutsu, Hideki; Nakajima, Ken-ichi; Tamada, Yasushi; Kojima, Katsura

    2014-01-01

    Spider dragline silk is a natural fiber that has excellent tensile properties; however, it is difficult to produce artificially as a long, strong fiber. Here, the spider (Araneus ventricosus) dragline protein gene was cloned and a transgenic silkworm was generated, that expressed the fusion protein of the fibroin heavy chain and spider dragline protein in cocoon silk. The spider silk protein content ranged from 0.37 to 0.61% w/w (1.4-2.4 mol%) native silkworm fibroin. Using a good silk-producing strain, C515, as the transgenic silkworm can make the raw silk from its cocoons for the first time. The tensile characteristics (toughness) of the raw silk improved by 53% after the introduction of spider dragline silk protein; the improvement depended on the quantity of the expressed spider dragline protein. To demonstrate the commercial feasibility for machine reeling, weaving, and sewing, we used the transgenic spider silk to weave a vest and scarf; this was the first application of spider silk fibers from transgenic silkworms. PMID:25162624

  18. Large-scale production of bioactive recombinant human acidic fibroblast growth factor in transgenic silkworm cocoons

    NASA Astrophysics Data System (ADS)

    Wang, Feng; Wang, Riyuan; Wang, Yuancheng; Zhao, Ping; Xia, Qingyou

    2015-11-01

    With an increasing clinical demand for functional therapeutic proteins every year, there is an increasing requirement for the massive production of bioactive recombinant human acidic fibroblast growth factor (r-haFGF). In this present study, we delicately explore a strategy for the mass production of r-haFGF protein with biological activity in the transgenic silkworm cocoons. The sequence-optimized haFGF was inserted into an enhanced sericin-1 expression system to generate the original transgenic silkworm strain, which was then further crossed with a PIG jumpstarter strain to achieve the remobilization of the expression cassette to a “safe harbor” locus in the genome for the efficient expression of r-haFGF. In consequence, the expression of r-haFGF protein in the mutant line achieved a 5.6-fold increase compared to the original strain. The high content of r-haFGF facilitated its purification and large-scald yields. Furthermore, the r-haFGF protein bioactively promoted the growth, proliferation and migration of NIH/3T3 cells, suggesting the r-haFGF protein possessed native mitogenic activity and the potential for wound healing. These results show that the silk gland of silkworm could be an efficient bioreactor strategy for recombinant production of bioactive haFGF in silkworm cocoons.

  19. Intestinal microecology associated with fluoride resistance capability of the silkworm (Bombyx mori L.).

    PubMed

    Li, Guan-Nan; Xia, Xue-Juan; Tang, Wen-Chao; Zhu, Yong

    2016-08-01

    The silkworm (Bombyx mori L.) is an ideal model of Lepidoptera. However, the diversity and function of the intestinal microbiota in the gut of silkworm remain largely unknown. Changes in the intestinal microecology in fluoride-resistant strain T6 and fluoride-susceptible strain 734 of the silkworm in response to fluoride exposure were investigated. T6 and 734 were treated with 200 mg/kg fluoride (designated as T6-T and 734-T groups) and deionized water (designated as T6-C and 734-C groups). Culture-dependent approach revealed that the numbers of intestinal bacteria in the 734-T group significantly decreased compared with that in the 734-C group (4.8 ± 0.6 × 10(7) CFU/mL vs. 7.5 ± 0.7 × 10(7) CFU/mL; P < 0.05). Analyses of the intestinal content pH showed that the pH decreased in the 734-T group only. Additionally, SCFA concentrations significantly decreased in both treatment groups compared with the control groups. High-throughput sequencing indicated that the intestinal microbiota in the 734-T group was significantly more diverse than those in the other groups. The bacterial community was composed of two dominant groups (Firmicutes and Proteobacteria). Principal component analyses revealed a significant difference in the composition of the intestinal microbiota in the 734-T group compared with those in the other groups. Thaumarchaeota and Euryarchaeota were more abundant in the 734-T group, but they were less abundant in the other groups. This study enhances our understanding about the diversity and function of silkworm intestinal microbiota in response to fluoride exposure among silkworm strains with diverse resistance.

  20. Construction and long term preservation of clonal transgenic silkworms using a parthenogenetic strain.

    PubMed

    Zabelina, Valeriya; Uchino, Keiro; Mochida, Yuji; Yonemura, Naoyuki; Klymenko, Vyacheslav; Sezutsu, Hideki; Tamura, Toshiki; Sehnal, František

    2015-10-01

    For the functional analysis of insect genes as well as for the production of recombinant proteins for biomedical use, clonal transgenic silkworms are very useful. We examined if they could be produced in the parthenogenetic strain that had been maintained for more than 40years as a female line in which embryogenesis is induced with nearly 100% efficiency by a heat shock treatment of unfertilized eggs. All individuals have identical female genotype. Silkworm transgenesis requires injection of the DNA constructs into the non-diapausing eggs at the preblastodermal stage of embryogenesis. Since our parthenogenetic silkworms produce diapausing eggs, diapause programing was eliminated by incubating ovaries of the parthenogenetic strain in standard male larvae. Chorionated eggs were dissected from the implants, activated by the heat shock treatment and injected with the transgene construct. Several transgenic individuals occurred in the daughter generation. Southern blotting analysis of two randomly chosen transgenic lines VTG1 and VTG14 revealed multiple transgene insertions. Insertions found in the parental females were transferred to the next generation without any changes in their sites and copy numbers, suggesting that transgenic silkworms can be maintained as clonal strains with homozygous transgenes. Cryopreservation was developed for the storage of precious genotypes. As shown for the VTG1 and VTG14 lines, larval ovaries can be stored in DMSO at the temperature of liquid nitrogen, transferred to Grace's medium during defrosting, and then implanted into larvae of either sex of the standard silkworm strains C146 and w1-pnd. Chorionated eggs, which developed in the implants, were dissected and activated by the heat shock to obtain females (nearly 100% efficiency) or by a cold shock to induce development to both sexes in 4% of the eggs. It was then possible to establish bisexual lines homozygous for the transgene. PMID:26112978

  1. 77 FR 23506 - Notice of Inventory Completion: The Region of Three Oaks Museum, Three Oaks, MI

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-19

    ... National Park Service Notice of Inventory Completion: The Region of Three Oaks Museum, Three Oaks, MI AGENCY: National Park Service, Interior. ACTION: Notice. SUMMARY: The Region of Three Oaks Museum has... contact The Region of Three Oaks Museum. Repatriation of the human remains to the Indian tribe...

  2. Identification and methods for prevention of Enterococcus mundtii infection in silkworm larvae, Bombyx mori, reared on artificial diet.

    PubMed

    Nwibo, Don Daniel; Matsumoto, Yasuhiko; Sekimizu, Kazuhisa

    2015-06-01

    Previously, it was reported that Enterococcus mundtii (E. mundtii) was associated with flacherie disease of silkworm larvae reared on artificial diet. In this study, we report that E. mundtii was isolated from diseased silkworm larvae, and validated as a pathogenic bacterium of the animal. When silkworm larva was infected with 1.04 × 10⁶ colony-forming units of E. mundtii via oral administration of diet, half population died within six days, indicating that the bacterium is pathogenic to silkworm. Less severe infection was found to cause anorexia and hamper the development of larvae. This pathogen was found to proliferate in both time- and dose-dependent manner in the gastrointestinal tract of the animal. The bacterium was isolated from powder of artificial diet made from mulberry leaves, and from mulberry leaves growing at a field. Minimum inhibitory concentration determination revealed that this bacterium was susceptible to tested antibiotics. Vancomycin treatment of diet significantly decreased the number of E. mundtii in intestine of silkworm larvae infected with the bacteria, compared to control. Furthermore, autoclaving or gamma ray irradiation of diet was also effective for exclusion of E. mundtii from the diet without the loss of its nutrient capacities. These results suggest that mulberry leaves used in making artificial diet for silkworm larvae is one of the sources of E. mundtii infection; and that antibiotic treatment, autoclaving or gamma ray irradiation of artificial diet can exclude the bacteria.

  3. Oxidative Damaged Products, Level of Hydrogen Peroxide, and Antioxidant Protection in Diapausing Pupa of Tasar Silk Worm, Antheraea mylitta: A Comparative Study in Two Voltine Groups.

    PubMed

    Sahoo, Alpana; Dandapat, Jagneshwar; Samanta, Luna

    2015-01-01

    The present study demonstrates tissue-specific (hemolymph and fat body) and inter-voltine [bivoltine (BV) and trivoltine (TV)] differences in oxidatively damaged products, H2O2 content, and the relative level of antioxidant protection in the diapausing pupae of Antheraea mylitta. Results suggest that fat body (FB) of both the voltine groups has oxidative predominance, as evident from the high value of lipid peroxidation and H2O2 content, despite better enzymatic defenses in comparison to hemolymph (HL). This may be attributed to the higher metabolic rate of the tissue concerned, concomitant with high lipid content and abundance of polyunsaturated fatty acids (PUFA). Nondetectable catalase activity in the pupal hemolymph of both strains apparently suggests an additional mechanism for H2O2 metabolism in the tissue. Inter-voltine comparison of the oxidative stress indices and antioxidant defense potential revealed that the TV group has a higher oxidative burden, lower activities for the antioxidant enzymes, and compensatory nonenzymatic protection from reduced glutathione and ascorbic acid.

  4. Molecular cloning, sequence analysis and expression of genome segment 7 (S7) of Antheraea mylitta cypovirus (AmCPV) that encodes a viral structural protein.

    PubMed

    Chavali, Venkata Ramana Murthy; Ghosh, Ananta K

    2007-10-01

    The Genome segment 7 (S7) of the 11 double stranded RNA genomes from Antheraea mylitta cypovirus (AmCPV) was converted to cDNA, cloned and sequenced. The nucleotide sequence showed that segment 7 consisted of 1789 nucleotides with an ORF of 530 amino acids and could encode a protein of approximately 61 kDa, termed P61. The 5' terminal sequence, AGTAAT and the 3' terminal sequence, AGAGC of the plus strand was found to be the same as genome segment 10 of AmCPV encoding polyhedrin. No sequence similarity was found by searching nucleic acid and protein sequence databases using BLAST. The secondary structure prediction showed the presence of 17 alpha-helices, 18 extended beta-sheets along the entire length of P61. The ORF of segment 7 was expressed in E. coli as His-tagged fusion protein, purified through Ni-NTA chromatography, and polyclonal antibody was raised in rabbit indicating that P61 is immunogenic. Immunoblot analysis using this antibody on viral infected cells as well as purified polyhedra showed that P61 is a viral structural protein. Motif scan search showed some similarity of P61 with Inosine monophosphate dehydrogenase (IMPDH) cystathionine-beta-synthase (CBS) domain at the C-terminus and it was hypothesized that by binding to single stranded viral RNA through its CBS domain P61 may help in virus replication or transcription.

  5. Inhibitors of Eicosanoid Biosynthesis Influencing the Transcripts Level of sHSP21.4 Gene Induced by Pathogen Infections, in Antheraea pernyi

    PubMed Central

    Zhang, Congfen; Dai, Lishang; Wang, Lei; Qian, Cen; Wei, Guoqing; Li, Jun; Zhu, Baojian; Liu, Chaoliang

    2015-01-01

    Small heat shock proteins (sHSPs) can regulate protein folding and protect cells from stress. To investigate the role of sHSPs in the silk-producing insect Antheraea pernyi response to microorganisms, a sHsp gene termed as Ap-sHSP21.4, was identified. This gene encoded a 21.4 kDa protein which shares the conserved structure of insect sHsps and belongs to sHSP21.4 family. Ap-sHSP21.4 was highly expressed in fat body and up-regulated in midgut and fat body of A. pernyi challenged with Escherichia coli, Beauveria bassiana and nuclear polyhedrosis virus (NPV), which was determined by quantitative real-time PCR. Meanwhile, knock down of Ap-sHSP21.4 with dsRNA result in the decrease at the expression levels of several immune response-related genes (defensin, Dopa decarboxylase, Toll1, lysozyme and Kazal-type serine protease inhibitor). Additionally, the impact of eicosanoid biosynthesis on the expression of Ap-sHSP21.4 response to NPV was determined using qPCR, inhibitors of eicosanoid biosynthesis significantly suppress Ap-HSP21.4 expression upon NPV challenge. All together, Ap-sHSP21.4 was involved in the immunity of A. pernyi against microorganism and possibly mediated by eicosanoids pathway. These results will shed light in the understanding of the pathogen-host interaction in A. pernyi. PMID:25844646

  6. Oxidative Damaged Products, Level of Hydrogen Peroxide, and Antioxidant Protection in Diapausing Pupa of Tasar Silk Worm, Antheraea mylitta: A Comparative Study in Two Voltine Groups.

    PubMed

    Sahoo, Alpana; Dandapat, Jagneshwar; Samanta, Luna

    2015-01-01

    The present study demonstrates tissue-specific (hemolymph and fat body) and inter-voltine [bivoltine (BV) and trivoltine (TV)] differences in oxidatively damaged products, H2O2 content, and the relative level of antioxidant protection in the diapausing pupae of Antheraea mylitta. Results suggest that fat body (FB) of both the voltine groups has oxidative predominance, as evident from the high value of lipid peroxidation and H2O2 content, despite better enzymatic defenses in comparison to hemolymph (HL). This may be attributed to the higher metabolic rate of the tissue concerned, concomitant with high lipid content and abundance of polyunsaturated fatty acids (PUFA). Nondetectable catalase activity in the pupal hemolymph of both strains apparently suggests an additional mechanism for H2O2 metabolism in the tissue. Inter-voltine comparison of the oxidative stress indices and antioxidant defense potential revealed that the TV group has a higher oxidative burden, lower activities for the antioxidant enzymes, and compensatory nonenzymatic protection from reduced glutathione and ascorbic acid. PMID:26816485

  7. Oak Ridge callibration recall program

    SciTech Connect

    Falter, K.G.; Wright, W.E.; Pritchard, E.W.

    1996-12-31

    A development effort was initiated within the Oak Ridge metrology community to address the need for a more versatile and user friendly tracking database that could be used across the Oak Ridge complex. This database, which became known as the Oak Ridge Calibration Recall Program (ORCRP), needed to be diverse enough for use by all three Oak Ridge facilities, as well as the seven calibration organizations that support them. Various practical functions drove the initial design of the program: (1) accessible by any user at any site through a multi-user interface, (2) real-time database that was able to automatically generate e-mail notices of due and overdue measuring and test equipment, (3) large memory storage capacity, and (4) extremely fast data access times. In addition, the program needed to generate reports on items such as instrument turnaround time, workload projections, and laboratory efficiency. Finally, the program should allow the calibration intervals to be modified, based on historical data. The developed program meets all of the stated requirements and is accessible over a network of computers running Microsoft Windows software.

  8. BmRobo2/3 is required for axon guidance in the silkworm Bombyx mori.

    PubMed

    Li, Xiao-Tong; Yu, Qi; Zhou, Qi-Sheng; Zhao, Xiao; Liu, Zhao-Yang; Cui, Wei-Zheng; Liu, Qing-Xin

    2016-02-15

    Axon guidance is critical for proper wiring of the nervous system. During the neural development, the axon guidance molecules play a key role and direct axons to choose the correct way to reach the target. Robo, as the receptor of axon guidance molecule Slit, is evolutionarily conserved from planarians to humans. However, the function of Robo in the silkworm, Bombyx mori, remained unknown. In this study, we cloned robo2/3 from B. mori (Bmrobo2/3), a homologue of robo2/3 in Tribolium castaneum. Moreover, BmRobo2/3 was localized in the neuropil, and RNAi-mediated knockdown of Bmrobo2/3 resulted in the longitudinal connectives forming closer to the midline. These data demonstrate that BmRobo2/3 is required for axon guidance in the silkworm.

  9. Proteomic-Based Insight into Malpighian Tubules of Silkworm Bombyx mori

    PubMed Central

    Liu, Shi-ping; Yi, Qi-ying; Hu, Cui-mei; Wang, Chen; Xia, Qing-you; Zhao, Ping

    2013-01-01

    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. PMID:24098719

  10. BmRobo2/3 is required for axon guidance in the silkworm Bombyx mori.

    PubMed

    Li, Xiao-Tong; Yu, Qi; Zhou, Qi-Sheng; Zhao, Xiao; Liu, Zhao-Yang; Cui, Wei-Zheng; Liu, Qing-Xin

    2016-02-15

    Axon guidance is critical for proper wiring of the nervous system. During the neural development, the axon guidance molecules play a key role and direct axons to choose the correct way to reach the target. Robo, as the receptor of axon guidance molecule Slit, is evolutionarily conserved from planarians to humans. However, the function of Robo in the silkworm, Bombyx mori, remained unknown. In this study, we cloned robo2/3 from B. mori (Bmrobo2/3), a homologue of robo2/3 in Tribolium castaneum. Moreover, BmRobo2/3 was localized in the neuropil, and RNAi-mediated knockdown of Bmrobo2/3 resulted in the longitudinal connectives forming closer to the midline. These data demonstrate that BmRobo2/3 is required for axon guidance in the silkworm. PMID:26625973

  11. Malpighi, Swammerdam and the colourful silkworm: replication and visual representation in early modern science.

    PubMed

    Cobb, Matthew

    2002-04-01

    In 1669, Malpighi published the first systematic dissection of an insect. The manuscript of this work contains a striking water-colour of the silkworm, which is described here for the first time. On repeating Malpighi's pioneering investigation, Swammerdam found what he thought were a number of errors, but was hampered by Malpighi's failure to explain his techniques. This may explain Swammerdam's subsequent description of his methods. In 1675, as he was about to abandon his scientific researches for a life of religious contemplation, Swammerdam destroyed his manuscript on the silkworm, but not before sending the drawings to Malpighi. These figures, with their rich and unique use of colour, are studied here for the first time. The role played by Henry Oldenburg, secretary of the Royal Society, in encouraging contact between the two men is emphasized and the way this exchange reveals the development of some key features of modern science - replication and modern scientific illustration - is discussed.

  12. Directly obtaining high strength silk fiber from silkworm by feeding carbon nanotubes.

    PubMed

    Wang, Jun-Ting; Li, Lu-Lu; Zhang, Meng-Yuan; Liu, Si-Lu; Jiang, Lin-Hai; Shen, Qing

    2014-01-01

    By feeding silkworm with the carbon nanotube, CNT, we directly obtained high strength silk fiber, SF, from silkworm. The CNT-based SF, SF/CNT, has a stress at 1.69GPa and a strain at about 24% both higher than those of the SF and are capable to compare with the super SF and even the spider fiber. Morphology comparison showed that the presence of CNT in SF caused the cross-section changed from triangle to ellipse. X-ray diffraction and infrared analysis indicated that the embedded CNT in SF caused an increase in silk-I structure. Specifically the amide-II structure reduced by about 5% and the amide-III structure increased by about 10%. Thermogravimetric analyses indicated that the presence of CNT in SF enhanced the thermal stability. Additionally, the presence of CNT in SF also enhanced the electrical property.

  13. Characterization of Transgenic Silkworm Yielded Biomaterials with Calcium-Binding Activity

    PubMed Central

    Wang, Shaohua; Zhang, Yuyu; Yang, Mingying; Ye, Lupeng; Gong, Lu; Qian, Qiujie; Shuai, Yajun; You, Zhengying; Chen, Yuyin; Zhong, Boxiong

    2016-01-01

    Silk fibers have many inherent properties that are suitable for their use in biomaterials. In this study, the silk fibroin was genetically modified by including a Ca-binding sequence, [(AGSGAG)6ASEYDYDDDSDDDDEWD]2 from shell nacreous matrix protein. It can be produced as fibers by transgenic silkworm. The Ca-binding activity and mineralization of the transgenic silk fibroin were examined in vitro. The results showed that this transgenic silk fibroin had relatively higher Ca-binding activity than unmodified silk fibroin. The increased Ca-binding activity could promote the usage of silk fibroin as a biomaterial in the pharmaceutical industry. This study shows the possibility of using silk fibroin as a mineralization accelerating medical material by generating genetically modified transgenic silkworm. PMID:27414647

  14. Toxicity of Bacillus thuringiensis crystal proteins against eri silkworm, Samia cynthia ricini (Lepidoptera: Saturniidae).

    PubMed

    Sandeep Kumar, Donthula; Tarakeswari, Muddanuru; Lakshminarayana, Maddukuri; Sujatha, Mulpuri

    2016-07-01

    Ten purified crystal proteins of Bacillus thuringiensis (Bt) were tested at concentrations ranging from 2.93 to 3000ng/cm(2) for their toxicity to eri silkworm through protein paint bioassays using castor leaves. Based on LC50 values, Cry1Aa (2.6ng/cm(2)) was highly toxic followed by Cry1Ac (29.3ng/cm(2)) and Cry1Ab (68.7ng/cm(2)). The Cry1Ca and Cry1Ea proteins were moderately toxic to eri silkworm larvae and resulted in 23% and 28% mortality, respectively at the highest concentration tested (3000ng/cm(2)). Only reduction in larval weight was observed with Cry2Aa, Cry1Da and Cry9Aa proteins while Cry3Aa and Cry1Ba proteins were found to be nontoxic. PMID:27377590

  15. Identification of a diazinon-metabolizing glutathione S-transferase in the silkworm, Bombyx mori

    PubMed Central

    Yamamoto, Kohji; Yamada, Naotaka

    2016-01-01

    The glutathione S-transferase superfamily play key roles in the metabolism of numerous xenobiotics. We report herein the identification and characterization of a novel glutathione S-transferase in the silkworm, Bombyx mori. The enzyme (bmGSTu2) conjugates glutathione to 1-chloro-2,4-dinitrobenzene, as well as metabolizing diazinon, one of the organophosphate insecticides. Quantitative reverse transcription–polymerase chain reaction analysis of transcripts demonstrated that bmGSTu2 expression was induced 1.7-fold in a resistant strain of B. mori. Mutagenesis of putative amino acid residues in the glutathione-binding site revealed that Ile54, Glu66, Ser67, and Asn68 are crucial for enzymatic function. These results provide insights into the catalysis of glutathione conjugation in silkworm by bmGSTu2 and into the detoxification of organophosphate insecticides. PMID:27440377

  16. Utilization of silkworm cocoon waste as a sorbent for the removal of oil from water.

    PubMed

    Moriwaki, Hiroshi; Kitajima, Shiori; Kurashima, Masahiro; Hagiwara, Ayaka; Haraguchi, Kazuma; Shirai, Koji; Kanekatsu, Rensuke; Kiguchi, Kenji

    2009-06-15

    The aim of this study is to investigate the utilization of silkworm cocoon waste, such as pierced or stained cocoons, as a sorbent material for the removal of motor and vegetable oils from water. The oil-sorption capacity, rate and reusability of the material were evaluated. The results show the high sorption capacity of the silkworm cocoon waste sorbent (42-52 g(oil)/g(sorbent) for motor oil and 37-60 g(oil)/g(sorbent) for vegetable oil). The oil sorbed onto the material could be recovered by squeezing the sorbent, and the squeezed material showed an oil-sorption capacity over 15 g(oil)/g(sorbent). We concluded that the material shows a high performance as a low cost and environmental friendly sorbent for the removal of oil from water.

  17. Characterization of Transgenic Silkworm Yielded Biomaterials with Calcium-Binding Activity.

    PubMed

    Wang, Shaohua; Zhang, Yuyu; Yang, Mingying; Ye, Lupeng; Gong, Lu; Qian, Qiujie; Shuai, Yajun; You, Zhengying; Chen, Yuyin; Zhong, Boxiong

    2016-01-01

    Silk fibers have many inherent properties that are suitable for their use in biomaterials. In this study, the silk fibroin was genetically modified by including a Ca-binding sequence, [(AGSGAG)6ASEYDYDDDSDDDDEWD]2 from shell nacreous matrix protein. It can be produced as fibers by transgenic silkworm. The Ca-binding activity and mineralization of the transgenic silk fibroin were examined in vitro. The results showed that this transgenic silk fibroin had relatively higher Ca-binding activity than unmodified silk fibroin. The increased Ca-binding activity could promote the usage of silk fibroin as a biomaterial in the pharmaceutical industry. This study shows the possibility of using silk fibroin as a mineralization accelerating medical material by generating genetically modified transgenic silkworm. PMID:27414647

  18. Identification of a diazinon-metabolizing glutathione S-transferase in the silkworm, Bombyx mori.

    PubMed

    Yamamoto, Kohji; Yamada, Naotaka

    2016-01-01

    The glutathione S-transferase superfamily play key roles in the metabolism of numerous xenobiotics. We report herein the identification and characterization of a novel glutathione S-transferase in the silkworm, Bombyx mori. The enzyme (bmGSTu2) conjugates glutathione to 1-chloro-2,4-dinitrobenzene, as well as metabolizing diazinon, one of the organophosphate insecticides. Quantitative reverse transcription-polymerase chain reaction analysis of transcripts demonstrated that bmGSTu2 expression was induced 1.7-fold in a resistant strain of B. mori. Mutagenesis of putative amino acid residues in the glutathione-binding site revealed that Ile54, Glu66, Ser67, and Asn68 are crucial for enzymatic function. These results provide insights into the catalysis of glutathione conjugation in silkworm by bmGSTu2 and into the detoxification of organophosphate insecticides. PMID:27440377

  19. [Identification and expression pattern of cathepsin family in silkworm (Bombyx mori)].

    PubMed

    Li, Yi; Zhou, Xiaoying; Li, Zhilang; Li, Jianwei; Chen, Shida; Guo, Chao; Hou, Yong; Zhao, Ping

    2015-12-01

    Silkworm is a holometabolous insect of Lepidoptera. During metamorphosis, significant morphological changes happen including the dissociation of old tissues and remodeling of new tissues. It has been reported that cathepsins are involved in these processes. Cathepsin is a kind of intracellular proteinase that exists in many species. It includes some subfamilies like cathepsin B, H and L. The studies on cathepsin are useful for clarifying the details of silkworm metamorphosis process. In total, 13 cathepsins were identified by screening the silkworm genome database. The basic information and the expression patterns about these genes were analyzed. Interestingly, an ovary-specific cathepsin L gene (Gene ID: BGIBMGAOO4622) was investigated by the data of silkworm microarray and real-time quantitative PCR (qPCR). The full-length cDNA is 1,209 bp, encoding a protein with 402 amino acids. Sequences alignment revealed that it has a high sequence similarity with cathepsin L of other species, and it is highly conserved in the active-site of the enzyme. The phylogenetic analysis showed that ovary-specific cathepsin L is clustered with other lepidopterous insects. Furthermore, this gene was cloned and prokaryotic expressed. Recombinant protein was present in inclusion body. Importantly, the qPCR result showed that the expression level of this gene is increasing during the early stage of pupal development and reaches the highest value at the 3rd day of pupal stage, which suggests that this gene may be involved in the process of development of the ovary and oocyte.

  20. A new arylalkylamine N-acetyltransferase in silkworm (Bombyx mori) affects integument pigmentation.

    PubMed

    Long, Yaohang; Li, Jiaorong; Zhao, Tianfu; Li, Guannan; Zhu, Yong

    2015-04-01

    Dopamine is a precursor for melanin synthesis. Arylalkylamine N-acetyltransferase (AANAT) is involved in the melatonin formation in insects because it could catalyze the transformation from dopamine to dopamine-N-acetyldopamine. In this study, we identified a new AANAT gene in the silkworm (Bombyx mori) and assessed its role in the silkworm. The cDNA of this gene encodes 233 amino acids that shares 57 % amino acid identity with the Bm-iAANAT protein. We thus refer to this gene as Bm-iAANAT2. To investigate the role of Bm-iAANAT2, we constructed a transgenic interference system using a 3xp3 promoter to suppress the expression of Bm-iAANAT2 in the silkworm. We observed that melanin deposition occurs in the head and integument in transgenic lines. To verify the melanism pattern, dopamine content and the enzyme activity of AANAT were determined by high-performance liquid chromatography (HPLC). We found that an increase in dopamine levels affects melanism patterns on the heads of transgenic B. mori. A reduction in the enzyme activity of AANAT leads to changes in dopamine levels. We analyzed the expression of the Bm-iAANAT2 genes by qPCR and found that the expression of Bm-iAANAT2 gene is significantly lower in transgenic lines. Our results lead us to conclude that Bm-iAANAT2 is a new arylalkylamine N-acetyltransferase gene in the silkworm and is involved in the metabolism of the dopamine to avoid the generation of melanin.

  1. Micro/Nanometer-scale fiber with highly ordered structures by mimicking the spinning process of silkworm.

    PubMed

    Chae, Su-Kyoung; Kang, Edward; Khademhosseini, Ali; Lee, Sang-Hoon

    2013-06-11

    A new method for the microfluidic spinning of ultrathin fibers with highly ordered structures is proposed by mimicking the spinning mechanism of silkworms. The self-aggregation is driven by dipole-dipole attractions between polar polymers upon contact with a low-polarity solvent to form fibers with nanostrands. The induction of Kelvin-Helmholtz instabilities at the dehydrating interface between two miscible fluids generates multi-scale fibers in a single microchannel.

  2. Effects of TiO2 nanoparticles on nutrition metabolism in silkworm fat body

    PubMed Central

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

    2016-01-01

    ABSTRACT Silkworm (Bombyx mori) is an important economic insect with a fat body that plays a crucial role in the storage and transfer of nutrients. It is also known that TiO2 nanoparticles (NPs) can improve feed efficiency and promote silk protein synthesis in the silkworm. In this study, we profiled gene expression in the silkworm fat body after TiO2 NP treatment, validated the major RNA-seq findings, and determined the contents of trehalose and triglyceride, the activity of lipase, and the amount of total proteins. RNA-seq analysis revealed that TiO2 NP treatment caused significant expression changes in 341 genes (P≤0.01), 138 of which were upregulated while the other 203 were downregulated. The expression levels of two target genes in the insulin signaling pathway and two protein metabolism-related target genes, three lipid metabolism-associated target genes, two carbohydrate metabolism related target genes and expression levels of seven heat shock protein genes were increased, and that of threonine dehydratase gene and fatty acid transport protein gene were decreased. The RNA-seq results of 16 genes were validated by quantitative real-time PCR. The lipase activity, content of trehalose, and amount of total proteins were elevated by 3.86-fold, 1.34-fold, and 1.21-fold, respectively, and the content of triglyceride was decreased by 0.94-fold after TiO2 NP treatment. These results indicated that TiO2 NPs activated the insulin signaling pathway, promoted the metabolism of protein, fat, and carbohydrate, and improved nutrition metabolism. Our study provides new support for the understanding of the beneficial effect of TiO2 NPs on silkworm nutrient metabolism. PMID:27185267

  3. Effects of TiO2 nanoparticles on nutrition metabolism in silkworm fat body.

    PubMed

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

    2016-01-01

    Silkworm (Bombyx mori) is an important economic insect with a fat body that plays a crucial role in the storage and transfer of nutrients. It is also known that TiO2 nanoparticles (NPs) can improve feed efficiency and promote silk protein synthesis in the silkworm. In this study, we profiled gene expression in the silkworm fat body after TiO2 NP treatment, validated the major RNA-seq findings, and determined the contents of trehalose and triglyceride, the activity of lipase, and the amount of total proteins. RNA-seq analysis revealed that TiO2 NP treatment caused significant expression changes in 341 genes (P≤0.01), 138 of which were upregulated while the other 203 were downregulated. The expression levels of two target genes in the insulin signaling pathway and two protein metabolism-related target genes, three lipid metabolism-associated target genes, two carbohydrate metabolism related target genes and expression levels of seven heat shock protein genes were increased, and that of threonine dehydratase gene and fatty acid transport protein gene were decreased. The RNA-seq results of 16 genes were validated by quantitative real-time PCR. The lipase activity, content of trehalose, and amount of total proteins were elevated by 3.86-fold, 1.34-fold, and 1.21-fold, respectively, and the content of triglyceride was decreased by 0.94-fold after TiO2 NP treatment. These results indicated that TiO2 NPs activated the insulin signaling pathway, promoted the metabolism of protein, fat, and carbohydrate, and improved nutrition metabolism. Our study provides new support for the understanding of the beneficial effect of TiO2 NPs on silkworm nutrient metabolism. PMID:27185267

  4. Genome-wide transcriptional response of silkworm (Bombyx mori) to infection by the microsporidian Nosema bombycis.

    PubMed

    Ma, Zhengang; Li, Chunfeng; Pan, Guoqing; Li, Zhihong; Han, Bing; Xu, Jinshan; Lan, Xiqian; Chen, Jie; Yang, Donglin; Chen, Quanmei; Sang, Qi; Ji, Xiaocun; Li, Tian; Long, Mengxian; Zhou, Zeyang

    2013-01-01

    Microsporidia have attracted much attention because they infect a variety of species ranging from protists to mammals, including immunocompromised patients with AIDS or cancer. Aside from the study on Nosema ceranae, few works have focused on elucidating the mechanism in host response to microsporidia infection. Nosema bombycis is a pathogen of silkworm pébrine that causes great economic losses to the silkworm industry. Detailed understanding of the host (Bombyx mori) response to infection by N. bombycis is helpful for prevention of this disease. A genome-wide survey of the gene expression profile at 2, 4, 6 and 8 days post-infection by N. bombycis was performed and results showed that 64, 244, 1,328, 1,887 genes were induced, respectively. Up to 124 genes, which are involved in basal metabolism pathways, were modulated. Notably, B. mori genes that play a role in juvenile hormone synthesis and metabolism pathways were induced, suggesting that the host may accumulate JH as a response to infection. Interestingly, N. bombycis can inhibit the silkworm serine protease cascade melanization pathway in hemolymph, which may be due to the secretion of serpins in the microsporidia. N. bombycis also induced up-regulation of several cellular immune factors, in which CTL11 has been suggested to be involved in both spore recognition and immune signal transduction. Microarray and real-time PCR analysis indicated the activation of silkworm Toll and JAK/STAT pathways. The notable up-regulation of antimicrobial peptides, including gloverins, lebocins and moricins, strongly indicated that antimicrobial peptide defense mechanisms were triggered to resist the invasive microsporidia. An analysis of N. bombycis-specific response factors suggested their important roles in anti-microsporidia defense. Overall, this study primarily provides insight into the potential molecular mechanisms for the host-parasite interaction between B. mori and N. bombycis and may provide a foundation for

  5. Genome-Wide Transcriptional Response of Silkworm (Bombyx mori) to Infection by the Microsporidian Nosema bombycis

    PubMed Central

    Pan, Guoqing; Li, Zhihong; Han, Bing; Xu, Jinshan; Lan, Xiqian; Chen, Jie; Yang, Donglin; Chen, Quanmei; Sang, Qi; Ji, Xiaocun; Li, Tian; Long, Mengxian; Zhou, Zeyang

    2013-01-01

    Microsporidia have attracted much attention because they infect a variety of species ranging from protists to mammals, including immunocompromised patients with AIDS or cancer. Aside from the study on Nosema ceranae, few works have focused on elucidating the mechanism in host response to microsporidia infection. Nosema bombycis is a pathogen of silkworm pébrine that causes great economic losses to the silkworm industry. Detailed understanding of the host (Bombyx mori) response to infection by N. bombycis is helpful for prevention of this disease. A genome-wide survey of the gene expression profile at 2, 4, 6 and 8 days post-infection by N. bombycis was performed and results showed that 64, 244, 1,328, 1,887 genes were induced, respectively. Up to 124 genes, which are involved in basal metabolism pathways, were modulated. Notably, B. mori genes that play a role in juvenile hormone synthesis and metabolism pathways were induced, suggesting that the host may accumulate JH as a response to infection. Interestingly, N. bombycis can inhibit the silkworm serine protease cascade melanization pathway in hemolymph, which may be due to the secretion of serpins in the microsporidia. N. bombycis also induced up-regulation of several cellular immune factors, in which CTL11 has been suggested to be involved in both spore recognition and immune signal transduction. Microarray and real-time PCR analysis indicated the activation of silkworm Toll and JAK/STAT pathways. The notable up-regulation of antimicrobial peptides, including gloverins, lebocins and moricins, strongly indicated that antimicrobial peptide defense mechanisms were triggered to resist the invasive microsporidia. An analysis of N. bombycis-specific response factors suggested their important roles in anti-microsporidia defense. Overall, this study primarily provides insight into the potential molecular mechanisms for the host-parasite interaction between B. mori and N. bombycis and may provide a foundation for

  6. CONSTRUCTION OF SILKWORM MIDGUT cDNA LIBRARY FOR SCREEN AND SEQUENCE ANALYSIS OF PERITROPHIC MEMBRANE PROTEIN GENES.

    PubMed

    Zhou, Yi-Jun; Xue, Bin; Li, Yang-Yang; Li, Fan-Chi; Ni, Min; Shen, Wei-De; Gu, Zhi-Ya; Li, Bing; Shen, Wei-De; Gu, Zhi-Ya; Li, Bing

    2016-01-01

    Silkworm is an important economic insect and the model species for Lepidoptera. The midgut of silkworm is an important physiological barrier, as its peritrophic membrane (PM) can resist pathogen invasion. In this study, a silkworm midgut cDNA library was constructed in order to identify silkworm PM genes. The capacity of the initial library was 6.92 × 10(6) pfu/ml, along with a recombination rate of 92.14% and a postamplification titer of 4.10 × 10(9) pfu/ml. Three silkworm PM protein genes were obtained by immunoscreening, two of which were chitin-binding protein (CBP) genes and one of which was a chitin deacetylase (CDA) gene as revealed by sequence analysis. Three genes were named BmCBP02, BmCBP13, and BmCDA17, and their ORF sizes are 678, 1,029, and 645 bp, respectively; all of them contain sequences of chitin-binding domains. Phylogenetic analysis indicated that BmCBP02 has the highest consensus with Mamestra configurata CBP at 61.0%; BmCBP13 has the highest consensus with Loxostege sticticalis PM CBP at 53.35%; BmCDA17 has the highest consensus with Helicoverpa armigera CDA5a at 70.83%. Tissue transcriptional analysis revealed that all three genes were specifically expressed in the midgut, and during the developmental process of fifth-instar silkworms, the transcription of all the genes showed an upward trend. This study laid a foundation for further studies on the functions of silkworm PM genes.

  7. Effects of altered catecholamine metabolism on pigmentation and physical properties of sclerotized regions in the silkworm melanism mutant.

    PubMed

    Qiao, Liang; Li, Yuanhao; Xiong, Gao; Liu, Xiaofan; He, Songzhen; Tong, Xiaoling; Wu, Songyuan; Hu, Hai; Wang, Rixin; Hu, Hongwei; Chen, Lushi; Zhang, Li; Wu, Jie; Dai, Fangyin; Lu, Cheng; Xiang, Zhonghuai

    2012-01-01

    Catecholamine metabolism plays an important role in the determination of insect body color and cuticle sclerotization. To date, limited research has focused on these processes in silkworm. In the current study, we analyzed the interactions between catecholamines and melanin genes and their effects on the pigmentation patterns and physical properties of sclerotized regions in silkworm, using the melanic mutant melanism (mln) silkworm strain as a model. Injection of β-alanine into mln mutant silkworm induced a change in catecholamine metabolism and turned its body color yellow. Further investigation of the catecholamine content and expression levels of the corresponding melanin genes from different developmental stages of Dazao-mln (mutant) and Dazao (wild-type) silkworm revealed that at the larval and adult stages, the expression patterns of melanin genes precipitated dopamine accumulation corresponding to functional loss of Bm-iAANAT, a repressive effect of excess NBAD on ebony, and upregulation of tan in the Dazao-mln strain. During the early pupal stage, dopamine did not accumulate in Dazao-mln, since upregulation of ebony and black genes led to conversion of high amounts of dopamine into NBAD, resulting in deep yellow cuticles. Scanning electron microscope analysis of a cross-section of adult dorsal plates from both wild-type and mutant silkworm disclosed the formation of different layers in Dazao-mln owing to lack of NADA, compared to even and dense layers in Dazao. Analysis of the mechanical properties of the anterior wings revealed higher storage modulus and lower loss tangent in Dazao-mln, which was closely associated with the altered catecholamine metabolism in the mutant strain. Based on these findings, we conclude that catecholamine metabolism is crucial for the color pattern and physical properties of cuticles in silkworm. Our results should provide a significant contribution to Lepidoptera cuticle tanning research.

  8. A Genome-Wide Survey for Host Response of Silkworm, Bombyx mori during Pathogen Bacillus bombyseptieus Infection

    PubMed Central

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

    2009-01-01

    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

  9. Toyama Kametaro and Vernon Kellogg: silkworm inheritance experiments in Japan, Siam, and the United States, 1900-1912.

    PubMed

    Onaga, Lisa

    2010-01-01

    Japanese agricultural scientist Toyama Kametaro's report about the Mendelian inheritance of silkworm cocoon color in Studies on the Hybridology of Insects (1906) spurred changes in Japanese silk production and thrust Toyama and his work into a scholarly exchange with American entomologist Vernon Kellogg. Toyama's work, based on research conducted in Japan and Siam, came under international scrutiny at a time when analyses of inheritance flourished after the "rediscovery" of Mendel's laws of heredity in 1900. The hybrid silkworm studies in Asia attracted the attention of Kellogg, who was concerned with how experimental biology would be used to study the causes of natural selection. He challenged Toyama's conclusions that Mendelism alone could explain the inheritance patterns of silkworm characters such as cocoon color because they had been subject to hundreds of years of artificial selection, or breeding. This examination of the intersection of Japanese sericulture and American entomology probes how practical differences in scientific interests, societal responsibilities, and silkworm materiality were negotiated throughout the processes of legitimating Mendelian genetics on opposite sides of the Pacific. The ways in which Toyama and Kellogg assigned importance to certain silkworm properties show how conflicting intellectual orientations arose in studies of the same organism. Contestation about Mendelism took place not just on a theoretical level, but the debate was fashioned through each scientist's rationale about the categorization of silkworm breeds and races and what counted as "natural". This further mediated the acceptability of the silkworm not as an experimental organism, but as an appropriately "natural" insect with which to demonstrate laws of inheritance. All these shed light on the challenges that came along with the use of agricultural animals to convincingly articulate new biological principles.

  10. Developmental Changes for the Hemolymph Metabolome of Silkworm (Bombyx mori L.).

    PubMed

    Zhou, Lihong; Li, Huihui; Hao, Fuhua; Li, Ning; Liu, Xin; Wang, Guoliang; Wang, Yulan; Tang, Huiru

    2015-05-01

    Silkworm (Bombyx mori) is a lepidopteran-holometabolic model organism. To understand its developmental biochemistry, we characterized the larval hemolymph metabonome from the third instar to prepupa stage using (1)H NMR spectroscopy whilst hemolymph fatty acid composition using GC-FID/MS. We unambiguously assigned more than 60 metabolites, among which tyrosine-o-β-glucuronide, mesaconate, homocarnosine, and picolinate were reported for the first time from the silkworm hemolymph. Phosphorylcholine was the most abundant metabolite in all developmental stages with exception for the periods before the third and fourth molting. We also found obvious developmental dependence for the hemolymph metabonome involving multiple pathways including protein biosyntheses, glycolysis, TCA cycle, the metabolisms of choline amino acids, fatty acids, purines, and pyrimidines. Most hemolymph amino acids had two elevations during the feeding period of the fourth instar and prepupa stage. Trehalose was the major blood sugar before day 8 of the fifth instar, whereas glucose became the major blood sugar after spinning. C16:0, C18:0 and its unsaturated forms were dominant fatty acids in hemolymph. The developmental changes of hemolymph metabonome were associated with dietary nutrient intakes, biosyntheses of cell membrane, pigments, proteins, and energy metabolism. These findings offered essential biochemistry information in terms of the dynamic metabolic changes during silkworm development.

  11. A novel Lozenge gene in silkworm, Bombyx mori regulates the melanization response of hemolymph.

    PubMed

    Xu, Man; Wang, Xue; Tan, Juan; Zhang, Kui; Guan, Xi; Patterson, Laurence H; Ding, Hanfei; Cui, Hongjuan

    2015-11-01

    Runt-related (RUNX) transcription factors are evolutionarily conserved either in vertebrate or invertebrate. Lozenge (Lz), a members of RUNX family as well as homologue of AML-1, functions as an important transcription factor regulating the hemocytes differentiation. In this paper, we identified and characterized RUNX family especially Lz in silkworm, which is a lepidopteran model insect. The gene expression analysis illustrated that BmLz was highly expressed in hemocytes throughout the whole development period, and reached a peak in glutonous stage. Over-expression of BmLz in silkworm accelerated the melanization process of hemolymph, and led to instantaneously up-regulation of prophenoloxidases (PPOs), which were key enzymes in the melanization process. Further down-regulation of BmLz expression by RNA interference resulted in the significant delay of melanization reaction of hemolymph. These findings suggested that BmLz regulated the melanization process of hemolymph by inducing PPOs expression, and played a critical role in innate immunity defense in silkworm.

  12. Characterization and identification of the integrin family in silkworm, Bombyx mori.

    PubMed

    Zhang, Kui; Xu, Man; Su, Jingjing; Yu, Shuang; Sun, Zhongfeng; Li, Yutian; Zhang, Weibo; Hou, Jianbing; Shang, Lijun; Cui, Hongjuan

    2014-10-01

    As an important economic insect, Bombyx mori is also a useful model organism for lepidopteran insect. Integrins are evolutionarily conserved from sponges to humans, and play vital roles in many physiological and pathological processes. To explore their diverse functions of integrins in insect, eleven integrins including six α and five β subunits were cloned and characterized from silkworm. Our results showed that integrins from silkworm own more family members compared to other invertebrates. Among those α subunits, integrins α1, α2, and the other four subunits belong to PS1, PS2, and PS3 groups, respectively. The β subunits mainly gather in the insect βν group except the β1 subunit which belongs to the insect β group. Expression profiles demonstrated that the integrins exhibited distinct patterns, but were mainly expressed in hemocytes. α1 and β2 subunits are the predominant ones either in the embryogenesis or larva stages. Interestingly, integrins were significantly up-regulated after stimulated by 20-hydroxyecdysone (20-E) in vivo. These results indicate that integrins perform diverse functions in hemocytes of silkworm. Overall, our results provide a new insight into the functional and evolutionary features of integrins.

  13. Molecular cloning, expression and characterization of acylpeptide hydrolase in the silkworm, Bombyx mori.

    PubMed

    Fu, Ping; Sun, Wei; Zhang, Ze

    2016-04-10

    Acylpeptide hydrolase (APH) can catalyze the release of the N-terminal amino acid from acetylated peptides. There were many documented examples of this enzyme in various prokaryotic and eukaryotic organisms. However, knowledge about APH in insects still remains unknown. In this study, we cloned and sequenced a putative silkworm Bombyx mori APH (BmAPH) gene. The BmAPH gene encodes a protein of 710 amino acids with a predicted molecular mass of 78.5kDa. The putative BmAPH and mammal APHs share about 36% amino acid sequence identity, yet key catalytic residues are conserved (Ser566, Asp654, and His686). Expression and purification of the recombinant BmAPH in Escherichia coli showed that it has acylpeptide hydrolase activity toward the traditional substrate, Ac-Ala-pNA. Furthermore, organophosphorus (OP) insecticides, chlorpyrifos, phoxim, and malathion, significantly inhibited the activity of the APH both in vitro and in vivo. In addition, BmAPH was expressed in all tested tissues and developmental stages of the silkworm. Finally, immunohistochemistry analysis showed that BmAPH protein was localized in the basement membranes. These results suggested that BmAPH may be involved in enhancing silkworm tolerance to the OP insecticides. In a word, our results provide evidence for understanding of the biological function of APH in insects.

  14. Aspartate Decarboxylase is Required for a Normal Pupa Pigmentation Pattern in the Silkworm, Bombyx mori

    PubMed Central

    Dai, Fangyin; Qiao, Liang; Cao, Cun; Liu, Xiaofan; Tong, Xiaoling; He, Songzhen; Hu, Hai; Zhang, Li; Wu, Songyuan; Tan, Duan; Xiang, Zhonghuai; Lu, Cheng

    2015-01-01

    The pigmentation pattern of Lepidoptera varies greatly in different development stages. To date, the effects of key genes in the melanin metabolism pathway on larval and adult body color are distinct, yet the effects on pupal pigmentation remains unclear. In the silkworm, Bombyx mori, the black pupa (bp) mutant is only specifically melanized at the pupal stage. Using positional cloning, we found that a mutation in the Aspartate decarboxylase gene (BmADC) is causative in the bp mutant. In the bp mutant, a SINE-like transposon with a length of 493 bp was detected ~2.2 kb upstream of the transcriptional start site of BmADC. This insertion causes a sharp reduction in BmADC transcript levels in bp mutants, leading to deficiency of β-alanine and N-β-alanyl dopamine (NBAD), but accumulation of dopamine. Following injection of β-alanine into bp mutants, the color pattern was reverted that of the wild-type silkworms. Additionally, melanic pupae resulting from knock-down of BmADC in the wild-type strain were obtained. These findings show that BmADC plays a crucial role in melanin metabolism and in the pigmentation pattern of the silkworm pupal stage. Finally, this study contributes to a better understanding of pupa pigmentation patterns in Lepidoptera. PMID:26077025

  15. Developmental Changes for the Hemolymph Metabolome of Silkworm (Bombyx mori L.).

    PubMed

    Zhou, Lihong; Li, Huihui; Hao, Fuhua; Li, Ning; Liu, Xin; Wang, Guoliang; Wang, Yulan; Tang, Huiru

    2015-05-01

    Silkworm (Bombyx mori) is a lepidopteran-holometabolic model organism. To understand its developmental biochemistry, we characterized the larval hemolymph metabonome from the third instar to prepupa stage using (1)H NMR spectroscopy whilst hemolymph fatty acid composition using GC-FID/MS. We unambiguously assigned more than 60 metabolites, among which tyrosine-o-β-glucuronide, mesaconate, homocarnosine, and picolinate were reported for the first time from the silkworm hemolymph. Phosphorylcholine was the most abundant metabolite in all developmental stages with exception for the periods before the third and fourth molting. We also found obvious developmental dependence for the hemolymph metabonome involving multiple pathways including protein biosyntheses, glycolysis, TCA cycle, the metabolisms of choline amino acids, fatty acids, purines, and pyrimidines. Most hemolymph amino acids had two elevations during the feeding period of the fourth instar and prepupa stage. Trehalose was the major blood sugar before day 8 of the fifth instar, whereas glucose became the major blood sugar after spinning. C16:0, C18:0 and its unsaturated forms were dominant fatty acids in hemolymph. The developmental changes of hemolymph metabonome were associated with dietary nutrient intakes, biosyntheses of cell membrane, pigments, proteins, and energy metabolism. These findings offered essential biochemistry information in terms of the dynamic metabolic changes during silkworm development. PMID:25825269

  16. Elementary research of the formation mechanism of sex-related fluorescent cocoon of silkworm, Bombyx mori.

    PubMed

    Xiaolong, Hu; Renyu, Xue; Guangli, Cao; Xing, Zhang; Yilin, Zhang; Xiaohua, Yu; Yuqing, Zhang; Chengliang, Gong

    2012-02-01

    To understand mechanisms for the difference of uptaking and transporting the pigments between the male and female in the silkworm, Bombyx mori strain of sex-related fluorescent cocoon, the fluorescent pigments in the midgut lumen, midgut, blood, silk glands and cocoon were analyzed with thin-layer chromatography, and showed that fluorescent colors of cocoons consisted with that of blood and silk glands. The different fluorescent colors of cocoons between the male and female may be mainly caused by the difference of accumulation and transportation for fluorescent pigments in the midgut and in the silk glands. Furthermore the midgut proteins were separated with Native-PAGE, and the proteins respectively recovered from three fluorescent regions presenting on a Native-PAGE gel for the female silkworms were determined using shotgun proteomics and mass spectrometry sequencing, of which 60, 40 and 18 proteins respectively from the region 1, 2 and 3 were identified. It was found that the several kinds of low molecular mass 30 kDa lipoproteins and the actins could be detected in all three regions, troponin, 30 kDa lipoprotein and 27 kDa glycoprotein precursor could be detected in the region 2 and 3, suggesting these proteins may be fluorescent pigments binding candidates proteins. Analysis of gene ontology indicated that the identified proteins in the three regions linked to the cellular component, molecular function, and biological process categories. These results provide a new clew to understand the formation mechanism of sex-related fluorescent cocoon of silkworm.

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

    PubMed

    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.

  18. Aspartate Decarboxylase is Required for a Normal Pupa Pigmentation Pattern in the Silkworm, Bombyx mori.

    PubMed

    Dai, Fangyin; Qiao, Liang; Cao, Cun; Liu, Xiaofan; Tong, Xiaoling; He, Songzhen; Hu, Hai; Zhang, Li; Wu, Songyuan; Tan, Duan; Xiang, Zhonghuai; Lu, Cheng

    2015-06-16

    The pigmentation pattern of Lepidoptera varies greatly in different development stages. To date, the effects of key genes in the melanin metabolism pathway on larval and adult body color are distinct, yet the effects on pupal pigmentation remains unclear. In the silkworm, Bombyx mori, the black pupa (bp) mutant is only specifically melanized at the pupal stage. Using positional cloning, we found that a mutation in the Aspartate decarboxylase gene (BmADC) is causative in the bp mutant. In the bp mutant, a SINE-like transposon with a length of 493 bp was detected ~2.2 kb upstream of the transcriptional start site of BmADC. This insertion causes a sharp reduction in BmADC transcript levels in bp mutants, leading to deficiency of β-alanine and N-β-alanyl dopamine (NBAD), but accumulation of dopamine. Following injection of β-alanine into bp mutants, the color pattern was reverted that of the wild-type silkworms. Additionally, melanic pupae resulting from knock-down of BmADC in the wild-type strain were obtained. These findings show that BmADC plays a crucial role in melanin metabolism and in the pigmentation pattern of the silkworm pupal stage. Finally, this study contributes to a better understanding of pupa pigmentation patterns in Lepidoptera.

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

    NASA Astrophysics Data System (ADS)

    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.

  20. Design and optimization of an experimental bioregenerative life support system with higher plants and silkworms

    NASA Astrophysics Data System (ADS)

    Hu, Enzhu; Bartsev, Sergey I.; Zhao, Ming; Liu, Professor Hong

    The conceptual scheme of an experimental bioregenerative life support system (BLSS) for planetary exploration was designed, which consisted of four elements - human metabolism, higher plants, silkworms and waste treatment. 15 kinds of higher plants, such as wheat, rice, soybean, lettuce, mulberry, et al., were selected as regenerative component of BLSS providing the crew with air, water, and vegetable food. Silkworms, which producing animal nutrition for crews, were fed by mulberry-leaves during the first three instars, and lettuce leaves last two instars. The inedible biomass of higher plants, human wastes and silkworm feces were composted into soil like substrate, which can be reused by higher plants cultivation. Salt, sugar and some household material such as soap, shampoo would be provided from outside. To support the steady state of BLSS the same amount and elementary composition of dehydrated wastes were removed periodically. The balance of matter flows between BLSS components was described by the system of algebraic equations. The mass flows between the components were optimized by EXCEL spreadsheets and using Solver. The numerical method used in this study was Newton's method.

  1. Developmental Changes for the Hemolymph Metabolome of Silkworm (Bombyx moriL.)

    PubMed Central

    Zhou, Lihong; Li, Huihui; Hao, Fuhua; Li, Ning; Liu, Xin; Wang, Guoliang; Wang, Yulan; Tang, Huiru

    2015-01-01

    Silkworm (Bombyx mori) is a lepidopteran-holometabolic model organism. To understand its developmental biochemistry, we characterized the larval hemolymph metabonome from the third instar to prepupa stage using 1H NMR spectroscopy whilst hemolymph fatty acid composition using GC-FID/MS. We unambiguously assigned more than 60 metabolites, among which tyrosine-o-β-glucuronide, mesaconate, homocarnosine, and picolinate were reported for the first time from the silkworm hemolymph. Phosphorylcholine was the most abundant metabolite in all developmental stages with exception for the periods before the third and fourth molting. We also found obvious developmental dependence for the hemolymph metabonome involving multiple pathways including protein biosyntheses, glycolysis, TCA cycle, the metabolisms of choline amino acids, fatty acids, purines, and pyrimidines. Most hemolymph amino acids had two elevations during the feeding period of the fourth instar and prepupa stage. Trehalose was the major blood sugar before day 8 of the fifth instar, whereas glucose became the major blood sugar after spinning. C16:0, C18:0 and its unsaturated forms were dominant fatty acids in hemolymph. The developmental changes of hemolymph metabonome were associated with dietary nutrient intakes, biosyntheses of cell membrane, pigments, proteins, and energy metabolism. These findings offered essential biochemistry information in terms of the dynamic metabolic changes during silkworm development. PMID:25825269

  2. Primed Immune Responses Triggered by Ingested Bacteria Lead to Systemic Infection Tolerance in Silkworms

    PubMed Central

    Miyashita, Atsushi; Takahashi, Shinji; Ishii, Kenichi; Sekimizu, Kazuhisa; Kaito, Chikara

    2015-01-01

    In the present study, we examined whether microorganisms collaterally ingested by insects with their food activate the innate immune system to confer systemic resistance against subsequent bacterial invasion. Silkworms orally administered heat-killed Pseudomonas aeruginosa cells showed resistance against intra-hemolymph infection by P. aeruginosa. Oral administration of peptidoglycans, cell wall components of P. aeruginosa, conferred protective effects against P. aeruginosa infection, whereas oral administration of lipopolysaccharides, bacterial surface components, did not. In silkworms orally administered heat-killed P. aeruginosa cells, P. aeruginosa growth was inhibited in the hemolymph, and mRNA amounts of the antimicrobial peptides cecropin A and moricin were increased in the hemocytes and fat body. Furthermore, the amount of paralytic peptide, an insect cytokine that activates innate immune reactions, was increased in the hemolymph of silkworms orally administered heat-killed P. aeruginosa cells. These findings suggest that insects sense bacteria present in their food by peptidoglycan recognition, which activates systemic immune reactions to defend the insects against a second round of infection. PMID:26107254

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

    SciTech Connect

    Kajikawa, Mizuho; Sasaki, Kaori; Wakimoto, Yoshitaro; Toyooka, Masaru; Motohashi, Tomoko; Shimojima, Tsukasa; Takeda, Shigeki; Park, Enoch Y.; Maenaka, Katsumi

    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.

  4. A novel Lozenge gene in silkworm, Bombyx mori regulates the melanization response of hemolymph.

    PubMed

    Xu, Man; Wang, Xue; Tan, Juan; Zhang, Kui; Guan, Xi; Patterson, Laurence H; Ding, Hanfei; Cui, Hongjuan

    2015-11-01

    Runt-related (RUNX) transcription factors are evolutionarily conserved either in vertebrate or invertebrate. Lozenge (Lz), a members of RUNX family as well as homologue of AML-1, functions as an important transcription factor regulating the hemocytes differentiation. In this paper, we identified and characterized RUNX family especially Lz in silkworm, which is a lepidopteran model insect. The gene expression analysis illustrated that BmLz was highly expressed in hemocytes throughout the whole development period, and reached a peak in glutonous stage. Over-expression of BmLz in silkworm accelerated the melanization process of hemolymph, and led to instantaneously up-regulation of prophenoloxidases (PPOs), which were key enzymes in the melanization process. Further down-regulation of BmLz expression by RNA interference resulted in the significant delay of melanization reaction of hemolymph. These findings suggested that BmLz regulated the melanization process of hemolymph by inducing PPOs expression, and played a critical role in innate immunity defense in silkworm. PMID:26164197

  5. Oak Ridge National Laboratory Review

    SciTech Connect

    Krause, C.; Pearce, J.; Zucker, A.

    1992-01-01

    This report presents brief descriptions of the following programs at Oak Ridge National Laboratory: The effects of pollution and climate change on forests; automation to improve the safety and efficiency of rearming battle tanks; new technologies for DNA sequencing; ORNL probes the human genome; ORNL as a supercomputer research center; paving the way to superconcrete made with polystyrene; a new look at supercritical water used in waste treatment; and small mammals as environmental monitors.

  6. Gene expression patterns in response to pathogen challenge and interaction with hemolin suggest that the Yippee protein of Antheraea pernyi is involved in the innate immune response.

    PubMed

    Sun, Yu; Dai, Lishang; Sun, Yuxuan; Wang, Lei; Qian, Cen; Wei, Guoqing; Zhu, Bao-Jian; Liu, Chao-Liang

    2016-07-01

    Yippee was first identified as a protein that physically interacts with the Hemolin protein of Hyalophora cecropia. In this study, we identified a gene with a 366bp open reading frame (ORF) that encodes a 121 amino acid protein containing a conserved Yippee domain. We named this gene Ap-Yippee (Yippee gene from Antheraea pernyi), and investigated the role of the protein in the host immune response. A recombinant Ap-Yippee protein was expressed in Escherichia coli cells, and polyclonal antibodies were produced against the recombinant protein. Real-time PCR and a Western blot analysis revealed that Ap-Yippee is expressed in the hemocytes, Malpighian tubules, midgut, silk gland, epidermis, and fat bodies of A. pernyi, with the highest expression level observed in Malpighian tubules. The fifth instar larvae of A. pernyi were challenged by injecting them with nucleopolyhedrovirus (AP-NPV), the Gram-negative bacterium E. coli, the Gram-positive bacterium Micrococcus luteus, or the entomopathogenic fungus, Beauveria bassiana. These challenges with diverse pathogens resulted in differential expression patterns of the protein. A knockdown of the Ap-Yippee gene by small interfering RNA (siRNA) transfection had a significant influence on the expression of the hemolin in the pupae which was confirmed by qRT-PCR and Western blot. Furthermore, a possible protein-protein interaction between Ap-Yippee and Hemolin was explored by Far-Western blotting. Therefore, our data suggest that the Ap-Yippee protein is involved in a pathway that regulates the immune response of insects. PMID:27261060

  7. Serotonin Receptor B May Lock the Gate of PTTH Release/Synthesis in the Chinese Silk Moth, Antheraea pernyi; A Diapause Initiation/Maintenance Mechanism?

    PubMed Central

    Takeda, Makio

    2013-01-01

    The release of prothoracicotropic hormone, PTTH, or its blockade is the major endocrine switch regulating the developmental channel either to metamorphosis or to pupal diapause in the Chinese silk moth, Antheraea pernyi. We have cloned cDNAs encoding two types of serotonin receptors (5HTRA and B). 5HTRA-, and 5HTRB-like immunohistochemical reactivities (-ir) were colocalized with PTTH-ir in two pairs of neurosecretory cells at the dorsolateral region of the protocerebrum (DL). Therefore, the causal involvement of these receptors was suspected in PTTH release/synthesis. The level of mRNA5HTRB responded to 10 cycles of long-day activation, falling to 40% of the original level before activation, while that of 5HTRA was not affected by long-day activation. Under LD 16:8 and 12:12, the injection of dsRNA5HTRB resulted in early diapause termination, whereas that of dsRNA5HTRA did not affect the rate of diapause termination. The injection of dsRNA5HTRB induced PTTH accumulation, indicating that 5HTRB binding suppresses PTTH synthesis also. This conclusion was supported pharmacologically; the injection of luzindole, a melatonin receptor antagonist, plus 5th inhibited photoperiodic activation under LD 16:8, while that of 5,7-DHT, induced emergence in a dose dependent fashion under LD 12:12. The results suggest that 5HTRB may lock the PTTH release/synthesis, maintaining diapause. This could also work as diapause induction mechanism. PMID:24223937

  8. Purification and molecular cloning of an inducible gram-negative bacteria-binding protein from the silkworm, Bombyx mori.

    PubMed

    Lee, W J; Lee, J D; Kravchenko, V V; Ulevitch, R J; Brey, P T

    1996-07-23

    A 50-kDa hemolymph protein, having strong affinity to the cell wall of Gram(-) bacteria, was purified from the hemolymph of the silkworm, Bombyx mori. The cDNA encoding this Gram(-) bacteria-binding protein (GNBP) was isolated from an immunized silkworm fat body cDNA library and sequenced. Comparison of the deduced amino acid sequence with known sequences revealed that GNBP contained a region displaying significant homology to the putative catalytic region of a group of bacterial beta-1,3 glucanases and beta-1,3-1,4 glucanases. Silkworm GNBP was also shown to have amino acid sequence similarity to the vertebrate lipopolysaccharide receptor CD14 and was recognized specifically by a polygonal anti-CD14 antibody. Northern blot analysis showed that GNBP was constitutively expressed in fat body, as well as in cuticular epithelial cells of naive silkworms. Intense transcription was, however, rapidly induced following a cuticular or hemoceolien bacterial challenge. An mRNA that hybridized with GNBP cDNA was also found in the l(2)mbn immunocompetent Drosophila cell line. These observations suggest that GNBP is an inducible acute phase protein implicated in the immune response of the silkworm and perhaps other insects.

  9. Silkworm eggs: An ideal model for studying the biological effects of low energy Ar + ion interaction in animals

    NASA Astrophysics Data System (ADS)

    Ling, Lin; Liu, Xuelan; Xu, Jiaping; You, Zhengying; Zhou, Jingbo

    2011-09-01

    The object of the current work was to study low energy Ar + ion beam interactions with silkworm eggs and thus provide further understanding of the mechanisms involved in ion bombardment-induced direct gene transfer into silkworm eggs. In this paper, using low-energy Ar + ion beam bombardment combined with piggyBac transposon, we developed a novel method to induce gene transfer in silkworm. Using bombardment conditions optimized for egg-incubation (25 keV with ion fluences of 800 × 2.6 × 10 15 ions/cm 2 in dry state under vacuum), vector pBac{3 × P3-EGFPaf} and helper plasmid pHA3pig were successfully transferred into the silkworm eggs. Our results obtained from by PCR assay and genomic Southern blotting analysis of the G1 generations provide evidence that low-energy ion beam can generate some craters that play a role in acting as pathways of exogenous DNA molecules into silkworm eggs.

  10. Isolation and characterization of lipase-producing bacteria in the intestine of the silkworm, Bombyx mori, reared on different forage.

    PubMed

    Feng, Wei; Wang, Xiao-Qiang; Zhou, Wei; Liu, Guang-Ying; Wan, Yong-Ji

    2011-01-01

    The silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), an oligophagous insect that mainly feeds on mulberry leaves, is susceptible to entomopathogen infection when reared with tricuspid cudrania leaves. A total of 56 dominant bacterial strains, classified into 12 phylotypes based on bacteriological properties and analysis of 16S rRNA genes, were isolated from the intestine of the fourth and fifth instar silkworm larvae. Ten and seven phylotypes exist in the intestine of the silkworm larvae reared with mulberry leaves and tricuspid cudrania leaves, respectively. Four of them are common in the intestine of the two treatment groups. By screening their lipolytic ability on a Rhodamine B agar plate, nine lipase-producing bacterial strains were obtained and classified into six genera, including Bacillus, Brevibacterium, Corynebacterium, Staphylococcus, Klebsiella, and Stenotrophomonas. Except for Stenotrophomonas, which is common in both, the other genera only exist in the intestine of the silkworm larvae fed with mulberry leaves. In addition, by culture and fermentation in vitro, the maximum cell density and lipase activity of lipase-producing bacteria were examined at about 48 hours. The results indicate that diet has a significant impact on the gut bacterial community, especially lipase-producing bacteria. We suggest that the difference of lipase-producing bacterial diversity might be related to disease resistance of the silkworm. PMID:22243438

  11. A novel granulocyte-specific α integrin is essential for cellular immunity in the silkworm Bombyx mori.

    PubMed

    Zhang, Kui; Tan, Juan; Xu, Man; Su, Jingjing; Hu, Renjian; Chen, Yibiao; Xuan, Fan; Yang, Rui; Cui, Hongjuan

    2014-12-01

    Haemocytes play crucial roles in immune responses and survival in insects. Specific cell markers have proven effective in clarifying the function and haematopoiesis of haemocytes. The silkworm Bombyx mori is a good model for studying insect haemocytes; however, little is known about haemocyte-specific markers or their functions in silkworm. In this study, we identified the α subunit of integrin, BmintegrinαPS3, as being specifically and highly expressed in silkworm haemocytes. Immunofluorescence analysis validated the specificity of BmintegrinαPS3 in larval granulocytes. Further analyses indicated that haemocytes dispersed from haematopoietic organs (HPOs) into the circulating haemolymph could differentiate into granulocytes. In addition, the processes of encapsulation and phagocytosis were controlled by larval granulocytes. Our work demonstrated that BmintegrinαPS3 could be used as a specific marker for granulocytes and could be applied to future molecular cell biology studies.

  12. [The Xanthine Oxidase Inhibitory Activity and Hypouricemic Effects of Crude Drugs Obtained from the Silkworm in Mice].

    PubMed

    Tanaka, Ryuichirou; Miyata, Yuuma; Minakuchi, Naoki; Murakami, Ayako; Sakazaki, Fumitoshi

    2015-01-01

    This study evaluated the effects of crude drugs obtained from the silkworm in mice with oxonic acid-induced hyperuricemia using xanthine oxidase inhibitory activity and plasma uric acid levels. The plasma uric acid level was analyzed using an improved HPLC with UV detection (HPLC-UV) method, which enabled high-sensitivity analysis of a microliter of plasma. Using this method, we evaluated natural products administered orally to the hypouricemic mice. The plasma uric acid level of mice administered a water-soluble extract from silkworm larvae with botrytis (used in traditional Chinese medicine to reduce wind, lower blood pressure, and change platelet coagulation) was significantly lower than in the control group 1, 2, and 3 h after treatment. In addition, water soluble extracts from a fungus (NBRC 31161) metabolite and silkworm pupae and larvae reduced the plasma uric acid levels in mice compared with the control group.

  13. [The Xanthine Oxidase Inhibitory Activity and Hypouricemic Effects of Crude Drugs Obtained from the Silkworm in Mice].

    PubMed

    Tanaka, Ryuichirou; Miyata, Yuuma; Minakuchi, Naoki; Murakami, Ayako; Sakazaki, Fumitoshi

    2015-01-01

    This study evaluated the effects of crude drugs obtained from the silkworm in mice with oxonic acid-induced hyperuricemia using xanthine oxidase inhibitory activity and plasma uric acid levels. The plasma uric acid level was analyzed using an improved HPLC with UV detection (HPLC-UV) method, which enabled high-sensitivity analysis of a microliter of plasma. Using this method, we evaluated natural products administered orally to the hypouricemic mice. The plasma uric acid level of mice administered a water-soluble extract from silkworm larvae with botrytis (used in traditional Chinese medicine to reduce wind, lower blood pressure, and change platelet coagulation) was significantly lower than in the control group 1, 2, and 3 h after treatment. In addition, water soluble extracts from a fungus (NBRC 31161) metabolite and silkworm pupae and larvae reduced the plasma uric acid levels in mice compared with the control group. PMID:26423873

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

    PubMed

    Chen, Jianqing; Shu, Tejun; Lv, Zhengbing; Nie, Zuoming; Chen, Jian; Chen, Hao; Yu, Wei; Gai, Qijing; Zhang, Yaozhou

    2014-01-01

    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.

  15. Purification and Functional Characterization of a Protein: Bombyx mori Human Growth Hormone Like Protein in Silkworm Pupa

    PubMed Central

    Lv, Zhengbing; Nie, Zuoming; Chen, Jian; Chen, Hao; Yu, Wei; Gai, Qijing; Zhang, Yaozhou

    2014-01-01

    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. PMID:25469649

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

    PubMed

    Sun, Zhenli; Lu, Yahong; Zhang, Hao; Kumar, Dhiraj; Liu, Bo; Gong, Yongchang; Zhu, Min; Zhu, Liyuan; Liang, Zi; Kuang, Sulan; Chen, Fei; Hu, Xiaolong; Cao, Guangli; Xue, Renyu; Gong, Chengliang

    2016-01-01

    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.

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

    PubMed Central

    Zhang, Hao; Kumar, Dhiraj; Liu, Bo; Gong, Yongchang; Zhu, Min; Zhu, Liyuan; Liang, Zi; Kuang, Sulan; Chen, Fei; Hu, Xiaolong; Cao, Guangli; Xue, Renyu; Gong, Chengliang

    2016-01-01

    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. PMID:26745627

  18. Conceptual design of a bioregenerative life support system containing crops and silkworms

    NASA Astrophysics Data System (ADS)

    Hu, Enzhu; Bartsev, Sergey I.; Liu, Hong

    2010-04-01

    This article summarizes a conceptual design of a bioregenerative life support system for permanent lunar base or planetary exploration. The system consists of seven compartments - higher plants cultivation, animal rearing, human habitation, water recovery, waste treatment, atmosphere management, and storages. Fifteen kinds of crops, such as wheat, rice, soybean, lettuce, and mulberry, were selected as main life support contributors to provide the crew with air, water, and vegetable food. Silkworms fed by crop leaves were designated to produce partial animal nutrition for the crew. Various physical-chemical and biological methods were combined to reclaim wastewater and solid waste. Condensate collected from atmosphere was recycled into potable water through granular activated carbon adsorption, iodine sterilization, and trace element supplementation. All grey water was also purified though multifiltration and ultraviolet sterilization. Plant residue, human excrement, silkworm feces, etc. were decomposed into inorganic substances which were finally absorbed by higher plants. Some meat, ingredients, as well as nitrogen fertilizer were prestored and resupplied periodically. Meanwhile, the same amount and chemical composition of organic waste was dumped to maintain the steady state of the system. A nutritional balanced diet was developed by means of the linear programming method. It could provide 2721 kcal of energy, 375.5 g of carbohydrate, 99.47 g of protein, and 91.19 g of fat per capita per day. Silkworm powder covered 12.54% of total animal protein intakes. The balance of material flows between compartments was described by the system of stoichiometric equations. Basic life support requirements for crews including oxygen, food, potable and hygiene water summed up to 29.68 kg per capita per day. The coefficient of system material closure reached 99.40%.

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

    PubMed

    Sun, Zhenli; Lu, Yahong; Zhang, Hao; Kumar, Dhiraj; Liu, Bo; Gong, Yongchang; Zhu, Min; Zhu, Liyuan; Liang, Zi; Kuang, Sulan; Chen, Fei; Hu, Xiaolong; Cao, Guangli; Xue, Renyu; Gong, Chengliang

    2016-01-01

    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. PMID:26745627

  20. Diversity analysis of Beauveria bassiana isolated from infected silkworm in southwest China based on molecular data and morphological features of colony.

    PubMed

    Wang, Jing-jie; Yang, Li; Qiu, Xin; Liu, Yong-gui; Zhou, Wei; Wan, Yong-Ji

    2013-07-01

    Beauveria bassiana is an important entomopathogenic fungus that not only often causes infection and epidemics of wild insects but some strains also show pathogenicity to the silkworm, Bombyx mori. The present study is about diversity of B. bassiana isolated from the silkworm in southwest China. Five strains of B. bassiana were isolated from infected silkworm. Two isolates, GXtr1009 and GXtr1010, were isolated from infected silkworms treated with two kinds of biological pesticides applied in Guangxi province, and three isolates, SCsk1006, YNsk1106 and GXsk1011, were collected from naturally infected silkworms from different geographical locations in Yunnan and Sichuan. All of the isolates showed highly similar conidia and conidial fructification, but the colony characteristics demonstrated great differences among the isolates. The ITS and 18S rDNA sequence analysis was sufficient to identify all five isolates as B. bassiana. However, the dendrogram, based on the ISSR data, produced two large genetic groups. GXtr1009 and GXtr1010 comprised one group, and SCsk1006, YNsk1106 and GXsk1011 converged in a different large group. The results suggested that, although all of these five B. bassiana strains were pathogenic to silkworms, strains of biological pesticides could be differentiated from strains of naturally infected silkworm via ISSR analysis. PMID:23417263

  1. Emergency preparedness at Oak Ridge National Laboratory, Oak Ridge, Tennessee

    SciTech Connect

    Skipper, M.N.

    1990-03-01

    Emergency preparedness for industry was commonly believed to be an essential responsibility on the part of management. Therefore, this study was conducted to research and accumulate information and data on emergency preparedness at Oak Ridge National Laboratory (ORNL). The objective of this study was to conduct a thorough evaluation of emergency preparedness knowledge among employees to determine if they were properly informed or if they needed more training. Also, this study was conducted to provide insight to management as to what their responsibility was concerning this training. To assess employee emergency preparedness knowledge, a questionnaire was developed and administered to 100 employees at ORNL. The data was analyzed using frequencies and percentages of response and was displayed through the use of graphs within the report. 22 refs., 22 figs.

  2. Systematic Identification and Characterization of Long Non-Coding RNAs in the Silkworm, Bombyx mori.

    PubMed

    Wu, Yuqian; Cheng, Tingcai; Liu, Chun; Liu, Duolian; Zhang, Quan; Long, Renwen; Zhao, Ping; Xia, Qingyou

    2016-01-01

    Long noncoding RNAs (lncRNAs) are emerging as important regulators in various biological processes. However, to date, no systematic characterization of lncRNAs has been reported in the silkworm Bombyx mori. In the present study, we generated eighteen RNA-seq datasets with relatively high depth. Using an in-house designed lncRNA identification pipeline, 11,810 lncRNAs were identified for 5,556 loci. Among these lncRNAs, 474 transcripts were intronic lncRNAs (ilncRNAs), 6,250 transcripts were intergenic lncRNAs (lincRNAs), and 5,086 were natural antisense lncRNAs (lncNATs). Compared with protein-coding mRNAs, silkworm lncRNAs are shorter in terms of full length but longer in terms of exon and intron length. In addition, lncRNAs exhibit a lower level of sequence conservation, more repeat sequences overlapped and higher tissue specificity than protein-coding mRNAs in the silkworm. We found that 69 lncRNA transcripts from 33 gene loci may function as miRNA precursors, and 104 lncRNA transcripts from 72 gene loci may act as competing endogenous RNAs (ceRNAs). In total, 49.47% of all gene loci (2,749/5,556) for which lncRNAs were identified showed sex-biased expression. Co-expression network analysis resulted in 19 modules, 12 of which revealed relatively high tissue specificity. The highlighted darkgoldenrod module was specifically associated with middle and posterior silk glands, and the hub lncRNAs within this module were co-expressed with proteins involved in translation, translocation, and secretory processes, suggesting that these hub lncRNAs may function as regulators of the biosynthesis, translocation, and secretion of silk proteins. This study presents the first comprehensive genome-wide analysis of silkworm lncRNAs and provides an invaluable resource for genetic, evolutionary, and genomic studies of B. mori.

  3. Systematic Identification and Characterization of Long Non-Coding RNAs in the Silkworm, Bombyx mori

    PubMed Central

    Wu, Yuqian; Cheng, Tingcai; Liu, Chun; Liu, Duolian; Zhang, Quan; Long, Renwen; Zhao, Ping; Xia, Qingyou

    2016-01-01

    Long noncoding RNAs (lncRNAs) are emerging as important regulators in various biological processes. However, to date, no systematic characterization of lncRNAs has been reported in the silkworm Bombyx mori. In the present study, we generated eighteen RNA-seq datasets with relatively high depth. Using an in-house designed lncRNA identification pipeline, 11,810 lncRNAs were identified for 5,556 loci. Among these lncRNAs, 474 transcripts were intronic lncRNAs (ilncRNAs), 6,250 transcripts were intergenic lncRNAs (lincRNAs), and 5,086 were natural antisense lncRNAs (lncNATs). Compared with protein-coding mRNAs, silkworm lncRNAs are shorter in terms of full length but longer in terms of exon and intron length. In addition, lncRNAs exhibit a lower level of sequence conservation, more repeat sequences overlapped and higher tissue specificity than protein-coding mRNAs in the silkworm. We found that 69 lncRNA transcripts from 33 gene loci may function as miRNA precursors, and 104 lncRNA transcripts from 72 gene loci may act as competing endogenous RNAs (ceRNAs). In total, 49.47% of all gene loci (2,749/5,556) for which lncRNAs were identified showed sex-biased expression. Co-expression network analysis resulted in 19 modules, 12 of which revealed relatively high tissue specificity. The highlighted darkgoldenrod module was specifically associated with middle and posterior silk glands, and the hub lncRNAs within this module were co-expressed with proteins involved in translation, translocation, and secretory processes, suggesting that these hub lncRNAs may function as regulators of the biosynthesis, translocation, and secretion of silk proteins. This study presents the first comprehensive genome-wide analysis of silkworm lncRNAs and provides an invaluable resource for genetic, evolutionary, and genomic studies of B. mori. PMID:26771876

  4. Identification of a bipartite nuclear localization signal in the silkworm Masc protein.

    PubMed

    Sugano, Yudai; Kokusho, Ryuhei; Ueda, Masamichi; Fujimoto, Masaru; Tsutsumi, Nobuhiro; Shimada, Toru; Kiuchi, Takashi; Katsuma, Susumu

    2016-07-01

    The silkworm Masculinizer (Masc) gene encodes a CCCH-tandem zinc finger protein that controls both masculinization and dosage compensation. Masc protein is a nuclear protein, but the mechanism underlying the transport of this protein into the nucleus has not yet been elucidated. Here, we identified a functional bipartite nuclear localization signal (NLS) located between residues 274 and 290 of the Masc protein. Sequence comparison revealed that this bipartite NLS is evolutionarily conserved in Masc proteins from other lepidopteran insects. Furthermore, we showed that the degree of nuclear localization is not associated with the masculinizing activity of the Masc protein. PMID:27277067

  5. Hydroforming Applications at Oak Ridge

    SciTech Connect

    bird, e.l.; ludtka, g.m.

    1999-03-10

    Hydroforming technology is a robust forming process that produces components with high precision and complexity. The goal of this paper is to present a brief description of the sheet hydroforming process with respect to the authors' experience and capabilities. Following the authors' discussion of the sheet-metal forming application, the tubular hydroforming process is described in the context of one of our technology development programs with an automotive industrial partner. After that is a summary of the tubular hydroforming advisor (expert system) development activity, which was a significant part of this overall program based on previous experience in developing a design and manufacturing support hydroforming advisor for the Oak Ridge Y-12 Plant's weapons-component manufacturing needs. Therefore, this paper is divided into three sections: (1) Hydroforming of Stainless Steel Parts, (2) Tubular Hydroforming, and (3) Components of a Tubular Hydroforming Advisor.

  6. Production of an active feline interferon in the cocoon of transgenic silkworms using the fibroin H-chain expression system

    SciTech Connect

    Kurihara, H. . E-mail: Hiroyuki_Kurihara@nts.toray.co.jp; Sezutsu, H.; Tamura, T.; Yamada, K.

    2007-04-20

    We constructed the fibroin H-chain expression system to produce recombinant proteins in the cocoon of transgenic silkworms. Feline interferon (FeIFN) was used for production and to assess the quality of the product. Two types of FeIFN fusion protein, each with N- and C-terminal sequences of the fibroin H-chain, were designed to be secreted into the lumen of the posterior silk glands. The expression of the FeIFN/H-chain fusion gene was regulated by the fibroin H-chain promoter domain. The transgenic silkworms introduced these constructs with the piggyBac transposon-derived vector, which produced the normal sized cocoons containing each FeIFN/H-chain fusion protein. Although the native-protein produced by transgenic silkworms have almost no antiviral activity, the proteins after the treatment with PreScission protease to eliminate fibroin H-chain derived N- and C-terminal sequences from the products, had very high antiviral activity. This H-chain expression system, using transgenic silkworms, could be an alternative method to produce an active recombinant protein and silk-based biomaterials.

  7. Comprehensive profiling of lysine acetylation suggests the widespread function is regulated by protein acetylation in the silkworm, Bombyx mori.

    PubMed

    Nie, Zuoming; Zhu, Honglin; Zhou, Yong; Wu, Chengcheng; Liu, Yue; Sheng, Qing; Lv, Zhengbing; Zhang, Wenping; Yu, Wei; Jiang, Caiying; Xie, Longfei; Zhang, Yaozhou; Yao, Juming

    2015-09-01

    Lysine acetylation in proteins is a dynamic and reversible PTM and plays an important role in diverse cellular processes. In this study, using lysine-acetylation (Kac) peptide enrichment coupled with nano HPLC/MS/MS, we initially identified the acetylome in the silkworms. Overall, a total of 342 acetylated proteins with 667 Kac sites were identified in silkworm. Sequence motifs analysis around Kac sites revealed an enrichment of Y, F, and H in the +1 position, and F was also enriched in the +2 and -2 positions, indicating the presences of preferred amino acids around Kac sites in the silkworm. Functional analysis showed the acetylated proteins were primarily involved in some specific biological processes. Furthermore, lots of nutrient-storage proteins, such as apolipophorin, vitellogenin, storage proteins, and 30 K proteins, were highly acetylated, indicating lysine acetylation may represent a common regulatory mechanism of nutrient utilization in the silkworm. Interestingly, Ser2 proteins, the coating proteins of larval silk, were found to contain many Kac sites, suggesting lysine acetylation may be involved in the regulation of larval silk synthesis. This study is the first to identify the acetylome in a lepidoptera insect, and expands greatly the catalog of lysine acetylation substrates and sites in insects.

  8. 2A self-cleaving peptide-based multi-gene expression system in the silkworm Bombyx mori.

    PubMed

    Wang, Yuancheng; Wang, Feng; Wang, Riyuan; Zhao, Ping; Xia, Qingyou

    2015-01-01

    Fundamental and applied studies of silkworms have entered the functional genomics era. Here, we report a multi-gene expression system (MGES) based on 2A self-cleaving peptide (2A), which regulates the simultaneous expression and cleavage of multiple gene targets in the silk gland of transgenic silkworms. First, a glycine-serine-glycine spacer (GSG) was found to significantly improve the cleavage efficiency of 2A. Then, the cleavage efficiency of six types of 2As with GSG was analyzed. The shortest porcine teschovirus-1 2A (P2A-GSG) exhibited the highest cleavage efficiency in all insect cell lines that we tested. Next, P2A-GSG successfully cleaved the artificial human serum albumin (66 kDa) linked with human acidic fibroblast growth factor (20.2 kDa) fusion genes and vitellogenin receptor fragment (196 kD) of silkworm linked with EGFP fusion genes, importantly, vitellogenin receptor protein was secreted to the outside of cells. Furthermore, P2A-GSG successfully mediated the simultaneous expression and cleavage of a DsRed and EGFP fusion gene in silk glands and caused secretion into the cocoon of transgenic silkworms using our sericin1 expression system. We predicted that the MGES would be an efficient tool for gene function research and innovative research on various functional silk materials in medicine, cosmetics, and other biomedical areas.

  9. MicroRNA-281 regulates the expression of ecdysone receptor (EcR) isoform B in the silkworm, Bombyx mori

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hundreds of Bombyx mori miRNAs had been identified in recent years, but their function in vivo remains poorly understood. The silkworm EcR gene (BmEcR) has three transcriptional isoforms, A, B1 and B2. Isoform sequences are different in the 3’UTR region of the gene, which is the case only in insects...

  10. An efficient strategy for producing a stable, replaceable, highly efficient transgene expression system in silkworm, Bombyx mori.

    PubMed

    Long, Dingpei; Lu, Weijian; Zhang, Yuli; Bi, Lihui; Xiang, Zhonghuai; Zhao, Aichun

    2015-01-01

    We developed an efficient strategy that combines a method for the post-integration elimination of all transposon sequences, a site-specific recombination system, and an optimized fibroin H-chain expression system to produce a stable, replaceable, highly efficient transgene expression system in the silkworm (Bombyx mori) that overcomes the disadvantages of random insertion and post-integration instability of transposons. Here, we generated four different transgenic silkworm strains, and of one the transgenic strains, designated TS1-RgG2, with up to 16% (w/w) of the target protein in the cocoons, was selected. The subsequent elimination of all the transposon sequences from TS1-RgG2 was completed by the heat-shock-induced expression of the transposase in vivo. The resulting transgenic silkworm strain was designated TS3-g2 and contained only the attP-flanked optimized fibroin H-chain expression cassette in its genome. A phiC31/att-system-based recombinase-mediated cassette exchange (RMCE) method could be used to integrate other genes of interest into the same genome locus between the attP sites in TS3-g2. Controlling for position effects with phiC31-mediated RMCE will also allow the optimization of exogenous protein expression and fine gene function analyses in the silkworm. The strategy developed here is also applicable to other lepidopteran insects, to improve the ecological safety of transgenic strains in biocontrol programs.

  11. A composite method for mapping quantitative trait loci without interference of female achiasmatic and gender effects in silkworm, Bombyx mori.

    PubMed

    Li, C; Zuo, W; Tong, X; Hu, H; Qiao, L; Song, J; Xiong, G; Gao, R; Dai, F; Lu, C

    2015-08-01

    The silkworm, Bombyx mori, is an economically important insect that was domesticated more than 5000 years ago. Its major economic traits focused on by breeders are quantitative traits, and an accurate and efficient QTL mapping method is necessary to explore their genetic architecture. However, current widely used QTL mapping models are not well suited for silkworm because they ignore female achiasmate and gender effects. In this study, we propose a composite method combining rational population selection and special mapping methods to map QTL in silkworm. By determining QTL for cocoon shell weight (CSW), we demonstrated the effectiveness of this method. In the CSW mapping process, only 56 markers were used and five loci or chromosomes were detected, more than in previous studies. Additionally, loci on chromosomes 1 and 11 dominated and accounted for 35.10% and 15.03% of the phenotypic variance respectively. Unlike previous studies, epistasis was detected between loci on chromosomes 11 and 22. These mapping results demonstrate the power and convenience of this method for QTL mapping in silkworm, and this method may inspire the development of similar approaches for other species with special genetic characteristics.

  12. Nonfunctional ingestion of plant miRNAs in silkworm revealed by digital droplet PCR and transcriptome analysis

    PubMed Central

    Jia, Ling; Zhang, Dayan; Xiang, Zhonghuai; He, Ningjia

    2015-01-01

    Since a plant miRNA (miR168) cross-regulating a mammalian transcript was reported, miRNA-mediated cross-kingdom communication has become one of the most compelling but controversial topics. In the present study, we used silkworm and mulberry, which is a model for studies on the interactions between the insect and its host plant, to address whether miRNA-mediated cross-kingdom communication is a common phenomenon. The results of TA clone, Sanger sequencing and droplet digital PCR demonstrated that several mulberry-derived miRNAs could enter to silkworm hemolymph and multiple tested tissues. Synthetic miR166b was also detected in hemolymph and fat body. However, the ingestion of synthetic miR166b did not play roles in silkworm physiological progress, which was revealed by RNA-seq analyses, RT-PCR, and phenotypic investigations. Mulberry miRNAs are convincingly transferred to the silkworm orally and no physiological process associated with the miRNAs was demonstrable. The results provided a new aspect of cross-kingdom miRNA transfer. PMID:26195298

  13. Serine protease P-IIc is responsible for the digestion of yolk proteins at the late stage of silkworm embryogenesis.

    PubMed

    Wang, Dandan; Zhang, Yan; Dong, Zhaoming; Guo, Pengchao; Ma, Sanyuan; Guo, Kaiyu; Xia, Qingyou; Zhao, Ping

    2016-07-01

    In silkworms, yolk proteins comprise vitellin, egg-specific protein and 30K proteins, which are sequentially degraded by endogenous proteases strictly regulated during embryogenesis. Although the process has been extensively investigated, there is still a gap in the knowledge about the degradation of silkworm yolk proteins on the last two days of embryonic development. In the present study, we isolated and purified a gut serine protease P-IIc, which demonstrated optimal activity at 25 °C and pH 11. Semi-quantitative RT-PCR combined with western blotting showed that P-IIc was actively expressed and significantly accumulated in the gut on the last two days of embryogenesis. When natural yolk proteins were incubated with P-IIc in vitro, vitellin and ESP were selectively degraded. P-IIc also demonstrated activity towards 30K proteins as evidenced by rapid and complete digestion of BmLP1 and partial digestion of BmLP2 and BmLP3. Furthermore, RNAi knockdown of P-IIc in silkworm embryos significantly reduced the degradation rate of residual yolk proteins on embryonic day 10. Taken together, our results indicate that P-IIc represents an embryonic gut protease with a relatively broad substrate specificity, which plays an important role in the degradation of yolk proteins at the late stage of silkworm embryogenesis. PMID:27137459

  14. [Control effect and bacteriostasis of mulberry endophytic bacterium Burkholderia cepacia Lu10-1 on silkworm septicemia].

    PubMed

    Dong, Fa-Bao; Mu, Zhi-Mei; Yu, Qi; Zhao, Kai; Liu, Zhao-Yang; Wang, Yan-Wen; Gao, Hui-Ju

    2012-11-01

    A laboratory test was conducted to study the control effect and bacteriostasis of antagonistic bacterium Burkholderia cepacia Lu10-1 isolated from mulberry on silkworm septicemia, aimed to develop a new microbial pesticide to control silkworm diseases. The supernatant of Lu10-1 zymotic fluid achieved 41.2% control efficiency and 24.0% prophylactic effect on silkworm septicemia. The antibacterial crude extract of Lu10-1 had stronger antagonistic activity against Bacillus bombyseptieus. The diameter of inhibition zone reached 18.20 mm, and the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the antibacterial crude extract were 1.56 and 3.13 mg x mL(-1), respectively. After treated with the antibacterial crude extract, B. bombyseptieus never appeared logarithmic growth phase, its cell membrane permeability changed, intracellular protein leaked out, intracellular macromolecular protein degraded, and at last, the thalli cracked, inner substances out-flowed, cavity formed, and cell ablated. It was considered that the antagonistic substances of Lu10-1 strain could be used for controlling silkworm septicemia, with preferable development foreground. PMID:23431798

  15. An efficient strategy for producing a stable, replaceable, highly efficient transgene expression system in silkworm, Bombyx mori

    PubMed Central

    Long, Dingpei; Lu, Weijian; Zhang, Yuli; Bi, Lihui; Xiang, Zhonghuai; Zhao, Aichun

    2015-01-01

    We developed an efficient strategy that combines a method for the post-integration elimination of all transposon sequences, a site-specific recombination system, and an optimized fibroin H-chain expression system to produce a stable, replaceable, highly efficient transgene expression system in the silkworm (Bombyx mori) that overcomes the disadvantages of random insertion and post-integration instability of transposons. Here, we generated four different transgenic silkworm strains, and of one the transgenic strains, designated TS1-RgG2, with up to 16% (w/w) of the target protein in the cocoons, was selected. The subsequent elimination of all the transposon sequences from TS1-RgG2 was completed by the heat-shock-induced expression of the transposase in vivo. The resulting transgenic silkworm strain was designated TS3-g2 and contained only the attP-flanked optimized fibroin H-chain expression cassette in its genome. A phiC31/att-system-based recombinase-mediated cassette exchange (RMCE) method could be used to integrate other genes of interest into the same genome locus between the attP sites in TS3-g2. Controlling for position effects with phiC31-mediated RMCE will also allow the optimization of exogenous protein expression and fine gene function analyses in the silkworm. The strategy developed here is also applicable to other lepidopteran insects, to improve the ecological safety of transgenic strains in biocontrol programs. PMID:25739894

  16. Construction of a Baculovirus-Silkworm Multigene Expression System and Its Application on Producing Virus-Like Particles

    PubMed Central

    Su, Shuo; Yao, Ning; He, Jian; Peng, Li; Sun, Jingchen

    2012-01-01

    A new baculovirus-silkworm multigene expression system named Bombyx mori MultiBac is developed and described here, by which multiple expression cassettes can be introduced into the Bombyx mori nuclear polyhedrosis virus (BmNPV) genome efficiently. The system consists of three donor vectors (pCTdual, pRADM and pUCDMIG) and an invasive diaminopimelate (DAP) auxotrophic recipient E. coli containing BmNPV-Bacmid (BmBacmid) with a homologous recombination region, an attTn7 site and a loxp site. Two genes carried by pCTdual are firstly inserted into BmBacmid by homologous recombination, while the other eight genes in pRADM and pUCDMIG are introduced into BmBacmid through Tn7 transposition and cre-loxp recombination. Then the invasive and DAP auxotrophic E. coli carrying recombinant BmBacmid is directly injected into silkworm for expressing heterologous genes in larvae or pupae. Three structural genes of rotavirus and three fluorescent genes have been simultaneously expressed in silkworm larvae using our new system, resulting in the formation of virus-like particles (VLPs) of rotavirus and the color change of larvae. The VLPs were purified from hemolymph by ultracentrifugation using CsCl gradients, with a yield of 12.7 µg per larva. For the great capacity of foreign genes and the low cost of feeding silkworm, this high efficient BmMultiBac expression system provides a suitable platform to produce VLPs or protein complexes. PMID:22403668

  17. 2A self-cleaving peptide-based multi-gene expression system in the silkworm Bombyx mori

    PubMed Central

    Wang, Yuancheng; Wang, Feng; Wang, Riyuan; Zhao, Ping; Xia, Qingyou

    2015-01-01

    Fundamental and applied studies of silkworms have entered the functional genomics era. Here, we report a multi-gene expression system (MGES) based on 2A self-cleaving peptide (2A), which regulates the simultaneous expression and cleavage of multiple gene targets in the silk gland of transgenic silkworms. First, a glycine-serine-glycine spacer (GSG) was found to significantly improve the cleavage efficiency of 2A. Then, the cleavage efficiency of six types of 2As with GSG was analyzed. The shortest porcine teschovirus-1 2A (P2A-GSG) exhibited the highest cleavage efficiency in all insect cell lines that we tested. Next, P2A-GSG successfully cleaved the artificial human serum albumin (66 kDa) linked with human acidic fibroblast growth factor (20.2 kDa) fusion genes and vitellogenin receptor fragment (196 kD) of silkworm linked with EGFP fusion genes, importantly, vitellogenin receptor protein was secreted to the outside of cells. Furthermore, P2A-GSG successfully mediated the simultaneous expression and cleavage of a DsRed and EGFP fusion gene in silk glands and caused secretion into the cocoon of transgenic silkworms using our sericin1 expression system. We predicted that the MGES would be an efficient tool for gene function research and innovative research on various functional silk materials in medicine, cosmetics, and other biomedical areas. PMID:26537835

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

    PubMed

    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

  19. Resistance of transgenic silkworm to BmNPV could be improved by silencing ie-1 and lef-1 genes.

    PubMed

    Zhang, P; Wang, J; Lu, Y; Hu, Y; Xue, R; Cao, G; Gong, C

    2014-01-01

    RNA interference (RNAi)-mediated viral inhibition has been used in several organisms for improving viral resistance. In the present study, we reported the use of transgenic RNAi in preventing Bombyx mori nucleopolyhedrovirus (BmNPV) multiplication in the transgenic silkworm B. mori. We targeted the BmNPV immediate-early-1 (ie-1) and late expression factor-1 (lef-1) genes in the transiently transfected BmN cells, in the stable transformed BmN cell line and in the transgenic silkworms. We generated four piggyBac-based vectors containing short double-stranded ie-1 RNA (sdsie-1), short double-stranded lef-1 RNA (sdslef-1), long double-stranded ie-1 RNA (ldsie-1) and both sdsie-1 and sdslef-1 (sds-ie1-lef1) expression cassettes. Strong viral repression was observed in the transiently transfected cells and in the stable transformed BmN cells transfected with sds-ie-1, sdslef-1, ldsie-1 or sds-ie-lef. The decrease of ie-1 mRNA level in the sds-ie1-lef1 transiently transfected cells was most obvious among the cells transfected with different vectors. The inhibitory effect of viral multiplication was decreased in a viral dose-dependent manner; the infection ratio of transfected cells for sds-ie-1, sdslef-1, ldsie-1 and sds-ie-lef decreased by 18.83%, 13.73%, 6.93% and 30.63%, respectively, compared with control cells 5 days after infection. We generated transgenic silkworms using transgenic vector piggyantiIE-lef1-neo with sds-ie1-lef1 expression cassette; the fourth instar larvae of transgenic silkworms of generation G5 exhibited stronger resistance to BmNPV, the mortalities for the transgenic silkworms and control silkworms were 60% and 100%, respectively, at 11 days after inoculation with BmNPV (10(6) occlusion bodies per ml). These results suggest that double-stranded RNA expression of essential genes of BmNPV is a feasible method for breeding silkworms with a high antiviral capacity. PMID:24173242

  20. Midgut immune responses induced by bacterial infection in the silkworm, Bombyx mori.

    PubMed

    Zhang, Lei; Wang, Yan-wen; Lu, Zhi-qiang

    2015-10-01

    Insect gut epithelial cells produce reactive oxygen species (ROS) and antimicrobial peptides (AMPs) to protect hosts from pathogenic microorganisms. In this study, we evaluate the pathogenicity of Pseudomonas aeruginosa and Bacillus bombysepticus in the silkworm, Bombyx mori. Survival curves show that B. bombysepticus is deadly when larval silkworms are infected orally. Bacterial infection caused intestinal hydrogen peroxide (H2O2) and nitric oxide (NO) levels to increase significantly by 8 and 16 h post-infection (hpi), respectively. Real-time quantitative polymerase chain reaction (qPCR) analysis shows that the transcription levels of dual oxidase (Duox) and catalase (CAT) are highly up-regulated by P. aeruginosa infection at 8 hpi. P. aeruginosa infection induced nitric oxide synthase 2 (NOS2) expression at 16 hpi, which contributes to the generation of NO. mRNA levels of AMP genes, specifically Glovorin 2 and Glovorin 3, which obviously increase during the early infection stage. These results indicate that invading bacteria elevate intestinal ROS and NO levels and induce AMP gene transcription, which contributes to intestinal immune defense. PMID:26465135

  1. Midgut immune responses induced by bacterial infection in the silkworm, Bombyx mori.

    PubMed

    Zhang, Lei; Wang, Yan-wen; Lu, Zhi-qiang

    2015-10-01

    Insect gut epithelial cells produce reactive oxygen species (ROS) and antimicrobial peptides (AMPs) to protect hosts from pathogenic microorganisms. In this study, we evaluate the pathogenicity of Pseudomonas aeruginosa and Bacillus bombysepticus in the silkworm, Bombyx mori. Survival curves show that B. bombysepticus is deadly when larval silkworms are infected orally. Bacterial infection caused intestinal hydrogen peroxide (H2O2) and nitric oxide (NO) levels to increase significantly by 8 and 16 h post-infection (hpi), respectively. Real-time quantitative polymerase chain reaction (qPCR) analysis shows that the transcription levels of dual oxidase (Duox) and catalase (CAT) are highly up-regulated by P. aeruginosa infection at 8 hpi. P. aeruginosa infection induced nitric oxide synthase 2 (NOS2) expression at 16 hpi, which contributes to the generation of NO. mRNA levels of AMP genes, specifically Glovorin 2 and Glovorin 3, which obviously increase during the early infection stage. These results indicate that invading bacteria elevate intestinal ROS and NO levels and induce AMP gene transcription, which contributes to intestinal immune defense.

  2. Functional Loss of Bmsei Causes Thermosensitive Epilepsy in Contractile Mutant Silkworm, Bombyx mori.

    PubMed

    Nie, Hongyi; Cheng, Tingcai; Huang, Xiaofeng; Zhou, Mengting; Zhang, Yinxia; Dai, Fangyin; Mita, Kazuei; Xia, Qingyou; Liu, Chun

    2015-01-01

    The thermoprotective mechanisms of insects remain largely unknown. We reported the Bombyx mori contractile (cot) behavioral mutant with thermo-sensitive seizures phenotype. At elevated temperatures, the cot mutant exhibit seizures associated with strong contractions, rolling, vomiting, and a temporary lack of movement. We narrowed a region containing cot to ~268 kb by positional cloning and identified the mutant gene as Bmsei which encoded a potassium channel protein. Bmsei was present in both the cell membrane and cytoplasm in wild-type ganglia but faint in cot. Furthermore, Bmsei was markedly decreased upon high temperature treatment in cot mutant. With the RNAi method and injecting potassium channel blockers, the wild type silkworm was induced the cot phenotype. These results demonstrated that Bmsei was responsible for the cot mutant phenotype and played an important role in thermoprotection in silkworm. Meanwhile, comparative proteomic approach was used to investigate the proteomic differences. The results showed that the protein of Hsp-1 and Tn1 were significantly decreased and increased on protein level in cot mutant after thermo-stimulus, respectively. Our data provide insights into the mechanism of thermoprotection in insect. As cot phenotype closely resembles human epilepsy, cot might be a potential model for the mechanism of epilepsy in future.

  3. Cloning and analysis of DnaJ family members in the silkworm, Bombyx mori.

    PubMed

    Li, Yinü; Bu, Cuiyu; Li, Tiantian; Wang, Shibao; Jiang, Feng; Yi, Yongzhu; Yang, Huipeng; Zhang, Zhifang

    2016-01-15

    Heat shock proteins (Hsps) are involved in a variety of critical biological functions, including protein folding, degradation, and translocation and macromolecule assembly, act as molecular chaperones during periods of stress by binding to other proteins. Using expressed sequence tag (EST) and silkworm (Bombyx mori) transcriptome databases, we identified 27 cDNA sequences encoding the conserved J domain, which is found in DnaJ-type Hsps. Of the 27 J domain-containing sequences, 25 were complete cDNA sequences. We divided them into three types according to the number and presence of conserved domains. By analyzing the gene structures, intron numbers, and conserved domains and constructing a phylogenetic tree, we found that the DnaJ family had undergone convergent evolution, obtaining new domains to expand the diversity of its family members. The acquisition of the new DnaJ domains most likely occurred prior to the evolutionary divergence of prokaryotes and eukaryotes. The expression of DnaJ genes in the silkworm was generally higher in the fat body. The tissue distribution of DnaJ1 proteins was detected by western blotting, demonstrating that in the fifth-instar larvae, the DnaJ1 proteins were expressed at their highest levels in hemocytes, followed by the fat body and head. We also found that the DnaJ1 transcripts were likely differentially translated in different tissues. Using immunofluorescence cytochemistry, we revealed that in the blood cells, DnaJ1 was mainly localized in the cytoplasm.

  4. Expression, purification and characterization of an atypical 2-Cys peroxiredoxin from the silkworm, Bombyx mori.

    PubMed

    Zhang, L; Lu, Z

    2015-04-01

    Peroxiredoxins (Prxs) play important roles in protecting organisms against damage caused by reactive oxygen species (ROS). In this study, we cloned a cDNA of Bombyx mori peroxiredoxin 5 (BmPrx5), which contained a 565-bp open reading frame for a 188-residue protein. Sequence analysis indicated that BmPrx5 belongs to the atypical 2-Cys peroxiredoxin family. Recombinant BmPrx5 purified from Escherichia coli showed antioxidant activity that removes H2 O2 and protects DNA from oxidative damage. Quantitative real-time PCR showed that the level of BmPrx5 mRNA in haemocytes increased early and decreased by 24 h after injection of H2 O2 whereas, in the fat body, the transcript level decreased at 6 h and increased at 12 h. Pseudomonas aeruginosa and Staphylococcus aureus infection resulted in higher levels of H2 O2 in the haemolymph and of BmPrx5 mRNA in haemocytes at 8 h postinfection. These data suggest that BmPrx5 acts as an antioxidant enzyme to protect the silkworm from oxidative damage induced by bacterial infection. Further study is needed to elucidate the exact role of BmPrx5 in the silkworm immune system.

  5. Proteomics analysis of digestive juice from silkworm during Bombyx mori nucleopolyhedrovirus infection.

    PubMed

    Hu, Xiaolong; Zhu, Min; Wang, Simei; Zhu, Liyuan; Xue, Renyu; Cao, Guangli; Gong, Chengliang

    2015-08-01

    Previous studies have analyzed the midgut transcriptome and proteome after challenge with Bombyx mori nucleopolyhedrovirus (BmNPV), however little information is available on the digestive juice proteome after BmNPV challenge. This study investigated BmNPV infection-induced protein changes in the digestive juice of silkworms using shotgun proteomics and MS sequencing. From the digestive juice of normal third-day, fifth-instar silkworm larvae, 75 proteins were identified, 44 of which were unknown; from larvae 6 h after inoculation with BmNPV, 106 proteins were identified, of which 39 were unknown. After BmNPV challenge, more secreted proteins appeared that had antiviral and digestive features. GO annotation analysis clustered most proteins in the lumen into catalytic, binding, and metabolic processes. Numerous proteins were reported to have BmNPV interactions. Hsp70 protein cognate, lipase-1, and chlorophyllide A-binding protein precursor were upregulated significantly after BmNPV challenge. Levels of trypsin-like serine protease, beta-1,3-glucanase, catalase, and serine protease transcripts decreased or were not significantly change after BmNPV challenge. Taken together, these findings provided insights into the interaction between host and BmNPV and revealed potential functions of digestive juice after per os BmNPV infection.

  6. TRANSCRIPTION FACTOR Bmsage PLAYS A CRUCIAL ROLE IN SILK GLAND GENERATION IN SILKWORM, Bombyx mori.

    PubMed

    Xin, Hu-hu; Zhang, Deng-pan; Chen, Rui-ting; Cai, Zi-zheng; Lu, Yan; Liang, Shuang; Miao, Yun-gen

    2015-10-01

    Salivary gland secretion is altered in Drosophila embryos with loss of function of the sage gene. Saliva has a reduced volume and an increased electron density according to transmission electron microscopy, resulting in regions of tube dilation and constriction with intermittent tube closure. However, the precise functions of Bmsage in silkworm (Bombyx mori) are unknown, although its sequence had been deposited in SilkDB. From this, Bmsage is inferred to be a transcription factor that regulates the synthesis of silk fibroin and interacts with another silk gland-specific transcription factor, namely, silk gland factor-1. In this study, we introduced a germline mutation of Bmsage using the Cas9/sgRNA system, a genome-editing technology, resulting in deletion of Bmsage from the genome of B. mori. Of the 15 tested samples, seven displayed alterations at the target site. The mutagenesis efficiency was about 46.7% and there were no obvious off-target effects. In the screened homozygous mutants, silk glands developed poorly and the middle and posterior silk glands (MSG and PSG) were absent, which was significantly different from the wild type. The offspring of G0 mosaic silkworms had indel mutations causing 2- or 9-bp deletions at the target site, but exhibited the same abnormal silk gland structure. Mutant larvae containing different open-reading frames of Bmsage had the same silk gland phenotype. This illustrated that the mutant phenotype was due to Bmsage knockout. We conclude that Bmsage participates in embryonic development of the silk gland.

  7. Identification and characterization of three novel hemocyte-specific promoters in silkworm Bombyx mori.

    PubMed

    Zhang, Kui; Yu, Shuang; Su, Jingjing; Xu, Man; Tan, Peng; Zhang, Yajun; Xiang, Zhonghuai; Cui, Hongjuan

    2015-05-22

    Insect hemocytes play essential roles in the metabolism, metamorphosis and immunity, which are closely related events of growth and development. Here, four novel hemocyte-specific genes were obtained and conformed in our study, namely, Bmintβ2, Bmintβ3, BmCatO, and BmSw04862, respectively. Subsequently, their promoter sequences were cloned, and their activity in hemocytes, fat body, and silk gland were analyzed using recombinant AcNPV vector system in vivo. Our results showed that Bmintβ2, Bmintβ3, and BmCatO were hemocyte-specific promoters in the silkworm, Bombyx mori. Interestingly, Bmintβ2, and Bmintβ3 promoter regions were both located in their first intron. Further analysis of a series of BmCatO promoter truncations showed that a 254 bp region could function as a promoter element in the tissue-specificity expression. In summary, the results of this study revealed that we have identified three hemocyte-specific promoters in silkworm that will not only great significance for better understanding of hemocyte-specific gene, but also has potential applications in insect hematopoiesis and innate immunity research.

  8. Molecular and Physiological Characterization of Two Novel Multirepeat β-Thymosins from Silkworm, Bombyx mori.

    PubMed

    Ma, Shangshang; Kang, Zhiqiong; Lü, Peng; Yang, Yanhua; Yao, Qin; Xia, Hengchuan; Chen, Keping

    2015-01-01

    β-thymosin plays important roles in the development of the lymphatic system and the central nervous system in vertebrates. However, its role and function in invertebrates remain much less explored. Here, we firstly isolated a gene encoding β-thymosin in silkworm (Bombyx mori L.). Interestingly, this gene encodes two polypeptides, named as BmTHY1 and BmTHY2, via two different modes of RNA splicing. The recombinant proteins fused with an N-term GST tag were over-expressed in Escherichia coli (E. coli) and further purified to near homogenity to prepare mouse antibodies. The Western blot analysis showed that these proteins were expressed in various tissues and organs, as well as in different developmental stages. Amazingly, the expression of BmTHY2 was hugely increased during the pupae stage, indicating a specialized role in this period. The expression of these proteins was gradually decreased in BmN cells infected by BmNPV, suggesting they may play different roles in the virus infection. In addition, both BmTHY1 and BmTHY2 can interact with 14-3-3 of silkworm and Ubiquitin of BmNPV as shown by GST pull down and Co-IP assays, consistent with their roles in the regulation of the development of nervous system.

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

    PubMed

    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.

  10. Can the silkworm (Bombyx mori) be used as a human disease model?

    PubMed

    Tabunoki, Hiroko; Bono, Hidemasa; Ito, Katsuhiko; Yokoyama, Takeshi

    2016-02-01

    Bombyx mori (silkworm) is the most famous lepidopteran in Japan. B. mori has long been used in the silk industry and also as a model insect for agricultural research. In recent years, B. mori has attracted interest in its potential for use in pathological analysis of model animals. For example, the human macular carotenoid transporter was discovered using information of B. mori carotenoid transporter derived from yellow-cocoon strain. The B. mori carotenoid transport system is useful in human studies. To develop a human disease model, we characterized the human homologs of B. mori, and by constructing KAIKO functional annotation pipeline, and to analyze gene expression profile of a unique B. mori mutant strain using microarray analysis. As a result, we identified a novel molecular network involved in Parkinson's disease. Here we describe the potential use of a spontaneous mutant silkworm strain as a human disease model. We also summarize recent progress in the application of genomic information for annotation of human homologs in B. mori. The B. mori mutant will provide a clue to pathological mechanisms, and the findings will be helpful for the development of therapies and for medical drug discovery.

  11. Heritable genome editing with CRISPR/Cas9 in the silkworm, Bombyx mori.

    PubMed

    Wei, Wei; Xin, Huhu; Roy, Bhaskar; Dai, Junbiao; Miao, Yungen; Gao, Guanjun

    2014-01-01

    We report the establishment of an efficient and heritable gene mutagenesis method in the silkworm Bombyx mori using modified type II clustered regularly interspaced short palindromic repeats (CRISPR) with an associated protein (Cas9) system. Using four loci Bm-ok, BmKMO, BmTH, and Bmtan as candidates, we proved that genome alterations at specific sites could be induced by direct microinjection of specific guide RNA and Cas9-mRNA into silkworm embryos. Mutation frequencies of 16.7-35.0% were observed in the injected generation, and DNA fragments deletions were also noted. Bm-ok mosaic mutants were used to test for mutant heritability due to the easily determined translucent epidermal phenotype of Bm-ok-disrupted cells. Two crossing strategies were used. In the first, injected Bm-ok moths were crossed with wild-type moths, and a 28.6% frequency of germline mutation transmission was observed. In the second strategy, two Bm-ok mosaic mutant moths were crossed with each other, and 93.6% of the offsprings appeared mutations in both alleles of Bm-ok gene (compound heterozygous). In summary, the CRISPR/Cas9 system can act as a highly specific and heritable gene-editing tool in Bombyx mori. PMID:25013902

  12. A discovery of novel microRNAs in the silkworm (Bombyx mori) genome.

    PubMed

    Yu, Xiaomin; Zhou, Qing; Cai, Yimei; Luo, Qibin; Lin, Hongbin; Hu, Songnian; Yu, Jun

    2009-12-01

    MicroRNAs (miRNAs) are pivotal regulators involved in various physiological and pathological processes via their post-transcriptional regulation of gene expressions. We sequenced 14 libraries of small RNAs constructed from samples spanning the life cycle of silkworms, and discovered 50 novel miRNAs previously not known in animals and verified 43 of them using stem-loop RT-PCR. Our genome-wide analyses of 27 species-specific miRNAs suggest they arise from transposable elements, protein-coding genes duplication/transposition and random foldback sequences; which is consistent with the idea that novel animal miRNAs may evolve from incomplete self-complementary transcripts and become fixed in the process of co-adaptation with their targets. Computational prediction suggests that the silkworm-specific miRNAs may have a preference of regulating genes that are related to life-cycle-associated traits, and these genes can serve as potential targets for subsequent studies of the modulating networks in the development of Bombyx mori.

  13. CULTURE IN VITRO OF TISSUE FROM THE SILKWORM, BOMBYX MORI L

    PubMed Central

    Wyatt, S. Silver

    1956-01-01

    1. Ovarian tissue from Bombyx mori L. larvae about to pupate was cultured in Trager's (1935) salt solution and 10 per cent hemolymph, with indifferent results. Improvement of cultures was sought by modifying the culture medium. 2. To reduce the activity of the tyrosinase, hemolymph for culture medium was heated for 5 minutes at 60°C., and the coagulated protein removed. 3. A physiological solution was formulated containing cations and amino acids as they occur normally in silkworm hemolymph. In both hanging-drop and small tube cultures use of this medium brought about increased cell number, improved cell appearance, more rapid mitoses, and longer life of cultures. 4. To the solution formulated from analyses, tryptophan, cystine, cysteine, malate, fumarate, succinate, and α-ketoglutarate were added after testing individually, resulting in improved growth in cultures. 5. Use of a silkworm egg extract prepared 4 to 5 days after acid treatment produced an increase in cell number. 6. In small roller tube cultures, when the new medium was changed twice a week, the cells spread over the walls of the tube in 4 or 5 days (Figs. 8 and 9), rapid mitoses were observed after 2 weeks, and transparent active cells were present at 3 weeks. Subculturing was not attempted. PMID:13346039

  14. Advanced silk material spun by a transgenic silkworm promotes cell proliferation for biomedical application.

    PubMed

    Wang, Feng; Xu, Hanfu; Wang, Yuancheng; Wang, Riyuan; Yuan, Lin; Ding, Huan; Song, Chunnuan; Ma, Sanyuan; Peng, Zhixin; Peng, Zhangchuan; Zhao, Ping; Xia, Qingyou

    2014-12-01

    Natural silk fiber spun by the silkworm Bombyx mori is widely used not only for textile materials, but also for biofunctional materials. In the present study, we genetically engineered an advanced silk material, named hSFSV, using a transgenic silkworm, in which the recombinant human acidic fibroblast growth factor (hFGF1) protein was specifically synthesized in the middle silk gland and secreted into the sericin layer to surround the silk fiber using our previously optimized sericin1 expression system. The content of the recombinant hFGF1 in the hSFSV silk was estimated to be approximate 0.07% of the cocoon shell weight. The mechanical properties of hSFSV raw silk fiber were enhanced slightly compared to those of the wild-type raw silk fiber, probably due to the presence of the recombinant of hFGF1 in the sericin layer. Remarkably, the hSFSV raw silk significantly stimulated the cell growth and proliferation of NIH/3T3 mouse embryonic fibroblast cells, suggesting that the mitogenic activity of recombinant hFGF1 was well maintained and functioned in the sericin layer of hSFSV raw silk. These results show that the genetically engineered raw silk hSFSV could be used directly as a fine biomedical material for mass application. In addition, the strategy whereby functional recombinant proteins are expressed in the sericin layer of silk might be used to create more genetically engineered silks with various biofunctions and applications.

  15. Molecular and Physiological Characterization of Two Novel Multirepeat β-Thymosins from Silkworm, Bombyx mori

    PubMed Central

    Lü, Peng; Yang, Yanhua; Yao, Qin; Xia, Hengchuan; Chen, Keping

    2015-01-01

    β-thymosin plays important roles in the development of the lymphatic system and the central nervous system in vertebrates. However, its role and function in invertebrates remain much less explored. Here, we firstly isolated a gene encoding β-thymosin in silkworm (Bombyx mori L.). Interestingly, this gene encodes two polypeptides, named as BmTHY1 and BmTHY2, via two different modes of RNA splicing. The recombinant proteins fused with an N-term GST tag were over-expressed in Escherichia coli (E. coli) and further purified to near homogenity to prepare mouse antibodies. The Western blot analysis showed that these proteins were expressed in various tissues and organs, as well as in different developmental stages. Amazingly, the expression of BmTHY2 was hugely increased during the pupae stage, indicating a specialized role in this period. The expression of these proteins was gradually decreased in BmN cells infected by BmNPV, suggesting they may play different roles in the virus infection. In addition, both BmTHY1 and BmTHY2 can interact with 14-3-3 of silkworm and Ubiquitin of BmNPV as shown by GST pull down and Co-IP assays, consistent with their roles in the regulation of the development of nervous system. PMID:26474303

  16. Comparative Transcriptome Analysis Reveals Different Silk Yields of Two Silkworm Strains

    PubMed Central

    Li, Juan; Qin, Sheng; Yu, Huanjun; Zhang, Jing; Liu, Na; Yu, Ye; Hou, Chengxiang; Li, Muwang

    2016-01-01

    Cocoon and silk yields are the most important characteristics of sericulture. However, few studies have examined the genes that modulate these features. Further studies of these genes will be useful for improving the products of sericulture. JingSong (JS) and Lan10 (L10) are two strains having significantly different cocoon and silk yields. In the current study, RNA-Seq and quantitative polymerase chain reaction (qPCR) were performed on both strains in order to determine divergence of the silk gland, which controls silk biosynthesis in silkworms. Compared with L10, JS had 1375 differentially expressed genes (DEGs; 738 up-regulated genes and 673 down-regulated genes). Nine enriched gene ontology (GO) terms were identified by GO enrichment analysis based on these DEGs. KEGG enrichment analysis results showed that the DEGs were enriched in three pathways, which were mainly associated with the processing and biosynthesis of proteins. The representative genes in the enrichment pathways and ten significant DEGs were further verified by qPCR, the results of which were consistent with the RNA-Seq data. Our study has revealed differences in silk glands between the two silkworm strains and provides a perspective for understanding the molecular mechanisms determining silk yield. PMID:27159277

  17. Expression, purification and characterization of an atypical 2-Cys peroxiredoxin from the silkworm, Bombyx mori.

    PubMed

    Zhang, L; Lu, Z

    2015-04-01

    Peroxiredoxins (Prxs) play important roles in protecting organisms against damage caused by reactive oxygen species (ROS). In this study, we cloned a cDNA of Bombyx mori peroxiredoxin 5 (BmPrx5), which contained a 565-bp open reading frame for a 188-residue protein. Sequence analysis indicated that BmPrx5 belongs to the atypical 2-Cys peroxiredoxin family. Recombinant BmPrx5 purified from Escherichia coli showed antioxidant activity that removes H2 O2 and protects DNA from oxidative damage. Quantitative real-time PCR showed that the level of BmPrx5 mRNA in haemocytes increased early and decreased by 24 h after injection of H2 O2 whereas, in the fat body, the transcript level decreased at 6 h and increased at 12 h. Pseudomonas aeruginosa and Staphylococcus aureus infection resulted in higher levels of H2 O2 in the haemolymph and of BmPrx5 mRNA in haemocytes at 8 h postinfection. These data suggest that BmPrx5 acts as an antioxidant enzyme to protect the silkworm from oxidative damage induced by bacterial infection. Further study is needed to elucidate the exact role of BmPrx5 in the silkworm immune system. PMID:25512182

  18. In vitro studies on the structure and properties of silk fibroin aqueous solutions in silkworm.

    PubMed

    Jin, Yuan; Hang, Yichun; Luo, Jie; Zhang, Yaopeng; Shao, Huili; Hu, Xuechao

    2013-11-01

    The spinning process of silkworm in vivo attracts great attentions. In this work, the structures and properties of the silk fibroin (SF) aqueous solutions from different divisions of silk glands of silkworms were investigated by using polarized microscope, rotational rheometer, Raman spectrometer and dynamic laser light scattering instrument. It was found that only the anterior (A) division and the anterior part of middle division (MA) of silk gland showed birefringence. With flowing from the posterior part (MP) to the MA part in the middle division of silk gland, the SF aqueous solutions was gradually transformed from random coil/α-helix to β-sheet conformation. Meantime, the elastic and viscous nature of the SF aqueous solution changed, and the mean diameter of SF aggregates increased from 118 nm to 331 nm. It could be concluded that the structures and properties of the SF aqueous solutions were gradually changed along the silk gland and the liquid crystal structure was initially formed in the MA part of silk gland. PMID:23994738

  19. Participation of D-serine in the development and reproduction of the silkworm Bombyx mori.

    PubMed

    Tanigawa, Minoru; Suzuki, Chihiro; Niwano, Kimio; Kanekatsu, Rensuke; Tanaka, Hiroyuki; Horiike, Kihachiro; Hamase, Kenji; Nagata, Yoko

    2016-04-01

    The silkworm Bombyx mori contains high concentrations of free D-serine, an optical isomer of L-serine. To elucidate its function, we first investigated the localization of D-serine in various organs of silkworm larvae, pupae, and adult moths. Using immunohistochemical analysis with an anti-D-serine antibody, we found D-serine in the microvilli of midgut goblet and cylindrical cells and in peripheral matrix components of testicular and ovarian cells. By spectrophotometric analysis, D-serine was also found in the hemolymph and fat body. D-Alanine was not detected in the various organs by immunohistochemistry. Serine racemase, which catalyzes the inter-conversion of L- and D-serine, was found to co-localize with D-serine, and D-serine production from L-serine by intrinsic serine racemase was suggested. O-Phospho-L-serine is an inhibitor of serine racemase, and it was administered to the larvae to reduce the D-serine level. This reagent decreased the midgut caspase-3 level and caused a delay in spermatogenesis and oogenesis. The reagent also decreased mature sperm and egg numbers, suggesting D-serine participation in these processes. D-Serine administration induced an increase in pyruvate levels in testis, midgut, and fat body, indicating conversion of D-serine to pyruvate. On the basis of these results, together with our previous investigation of ATP biosynthesis in testis, we consider the possible involvement of D-serine in ATP synthesis for metamorphosis and reproduction.

  20. Cloning and analysis of DnaJ family members in the silkworm, Bombyx mori.

    PubMed

    Li, Yinü; Bu, Cuiyu; Li, Tiantian; Wang, Shibao; Jiang, Feng; Yi, Yongzhu; Yang, Huipeng; Zhang, Zhifang

    2016-01-15

    Heat shock proteins (Hsps) are involved in a variety of critical biological functions, including protein folding, degradation, and translocation and macromolecule assembly, act as molecular chaperones during periods of stress by binding to other proteins. Using expressed sequence tag (EST) and silkworm (Bombyx mori) transcriptome databases, we identified 27 cDNA sequences encoding the conserved J domain, which is found in DnaJ-type Hsps. Of the 27 J domain-containing sequences, 25 were complete cDNA sequences. We divided them into three types according to the number and presence of conserved domains. By analyzing the gene structures, intron numbers, and conserved domains and constructing a phylogenetic tree, we found that the DnaJ family had undergone convergent evolution, obtaining new domains to expand the diversity of its family members. The acquisition of the new DnaJ domains most likely occurred prior to the evolutionary divergence of prokaryotes and eukaryotes. The expression of DnaJ genes in the silkworm was generally higher in the fat body. The tissue distribution of DnaJ1 proteins was detected by western blotting, demonstrating that in the fifth-instar larvae, the DnaJ1 proteins were expressed at their highest levels in hemocytes, followed by the fat body and head. We also found that the DnaJ1 transcripts were likely differentially translated in different tissues. Using immunofluorescence cytochemistry, we revealed that in the blood cells, DnaJ1 was mainly localized in the cytoplasm. PMID:26434795

  1. Can the silkworm (Bombyx mori) be used as a human disease model?

    PubMed

    Tabunoki, Hiroko; Bono, Hidemasa; Ito, Katsuhiko; Yokoyama, Takeshi

    2016-02-01

    Bombyx mori (silkworm) is the most famous lepidopteran in Japan. B. mori has long been used in the silk industry and also as a model insect for agricultural research. In recent years, B. mori has attracted interest in its potential for use in pathological analysis of model animals. For example, the human macular carotenoid transporter was discovered using information of B. mori carotenoid transporter derived from yellow-cocoon strain. The B. mori carotenoid transport system is useful in human studies. To develop a human disease model, we characterized the human homologs of B. mori, and by constructing KAIKO functional annotation pipeline, and to analyze gene expression profile of a unique B. mori mutant strain using microarray analysis. As a result, we identified a novel molecular network involved in Parkinson's disease. Here we describe the potential use of a spontaneous mutant silkworm strain as a human disease model. We also summarize recent progress in the application of genomic information for annotation of human homologs in B. mori. The B. mori mutant will provide a clue to pathological mechanisms, and the findings will be helpful for the development of therapies and for medical drug discovery. PMID:26853920

  2. Homeodomain Protein Scr Regulates the Transcription of Genes Involved in Juvenile Hormone Biosynthesis in the Silkworm.

    PubMed

    Meng, Meng; Liu, Chun; Peng, Jian; Qian, Wenliang; Qian, Heying; Tian, Ling; Li, Jiarui; Dai, Dandan; Xu, Anying; Li, Sheng; Xia, Qingyou; Cheng, Daojun

    2015-11-02

    The silkworm Dominant trimolting (Moltinism, M³) mutant undergoes three larval molts and exhibits precocious metamorphosis. In this study, we found that compared with the wild-type (WT) that undergoes four larval molts, both the juvenile hormone (JH) concentration and the expression of the JH-responsive gene Krüppel homolog 1 (Kr-h1) began to be greater in the second instar of the M³ mutant. A positional cloning analysis revealed that only the homeodomain transcription factor gene Sex combs reduced (Scr) is located in the genomic region that is tightly linked to the M³ locus. The expression level of the Scr gene in the brain-corpora cardiaca-corpora allata (Br-CC-CA) complex, which controls the synthesis of JH, was very low in the final larval instar of both the M³ and WT larvae, and exhibited a positive correlation with JH titer changes. Importantly, luciferase reporter analysis and electrophoretic mobility shift assay (EMSA) demonstrated that the Scr protein could promote the transcription of genes involved in JH biosynthesis by directly binding to the cis-regulatory elements (CREs) of homeodomain protein on their promoters. These results conclude that the homeodomain protein Scr is transcriptionally involved in the regulation of JH biosynthesis in the silkworm.

  3. Molecular and Physiological Characterization of Two Novel Multirepeat β-Thymosins from Silkworm, Bombyx mori.

    PubMed

    Ma, Shangshang; Kang, Zhiqiong; Lü, Peng; Yang, Yanhua; Yao, Qin; Xia, Hengchuan; Chen, Keping

    2015-01-01

    β-thymosin plays important roles in the development of the lymphatic system and the central nervous system in vertebrates. However, its role and function in invertebrates remain much less explored. Here, we firstly isolated a gene encoding β-thymosin in silkworm (Bombyx mori L.). Interestingly, this gene encodes two polypeptides, named as BmTHY1 and BmTHY2, via two different modes of RNA splicing. The recombinant proteins fused with an N-term GST tag were over-expressed in Escherichia coli (E. coli) and further purified to near homogenity to prepare mouse antibodies. The Western blot analysis showed that these proteins were expressed in various tissues and organs, as well as in different developmental stages. Amazingly, the expression of BmTHY2 was hugely increased during the pupae stage, indicating a specialized role in this period. The expression of these proteins was gradually decreased in BmN cells infected by BmNPV, suggesting they may play different roles in the virus infection. In addition, both BmTHY1 and BmTHY2 can interact with 14-3-3 of silkworm and Ubiquitin of BmNPV as shown by GST pull down and Co-IP assays, consistent with their roles in the regulation of the development of nervous system. PMID:26474303

  4. Functional Loss of Bmsei Causes Thermosensitive Epilepsy in Contractile Mutant Silkworm, Bombyx mori

    NASA Astrophysics Data System (ADS)

    Nie, Hongyi; Cheng, Tingcai; Huang, Xiaofeng; Zhou, Mengting; Zhang, Yinxia; Dai, Fangyin; Mita, Kazuei; Xia, Qingyou; Liu, Chun

    2015-07-01

    The thermoprotective mechanisms of insects remain largely unknown. We reported the Bombyx mori contractile (cot) behavioral mutant with thermo-sensitive seizures phenotype. At elevated temperatures, the cot mutant exhibit seizures associated with strong contractions, rolling, vomiting, and a temporary lack of movement. We narrowed a region containing cot to ~268 kb by positional cloning and identified the mutant gene as Bmsei which encoded a potassium channel protein. Bmsei was present in both the cell membrane and cytoplasm in wild-type ganglia but faint in cot. Furthermore, Bmsei was markedly decreased upon high temperature treatment in cot mutant. With the RNAi method and injecting potassium channel blockers, the wild type silkworm was induced the cot phenotype. These results demonstrated that Bmsei was responsible for the cot mutant phenotype and played an important role in thermoprotection in silkworm. Meanwhile, comparative proteomic approach was used to investigate the proteomic differences. The results showed that the protein of Hsp-1 and Tn1 were significantly decreased and increased on protein level in cot mutant after thermo-stimulus, respectively. Our data provide insights into the mechanism of thermoprotection in insect. As cot phenotype closely resembles human epilepsy, cot might be a potential model for the mechanism of epilepsy in future.

  5. Sorbitol as an arrester of embryonic development in diapausing eggs of the silkworm, Bombyx mori.

    PubMed

    Horie; Kanda; Mochida

    2000-06-01

    Recently, it was confirmed that embryos derived from diapausing eggs of the silkworm, Bombyx mori, begin their development and reach larval maturity on mulberry leaves, when the naked eggs are cultured in vitro. In this study, we found that the method of embryo culture is useful for determining the physiological regulation of diapause. We show that the development of embryos derived from diapausing eggs was strongly inhibited by the addition of either sorbitol or trehalose to the culture medium. Furthermore, this inhibitory effect disappeared when the embryos were cultured in a control medium which did not contain either sorbitol or trehalose, indicating that the inhibitory reactions caused by both substances are reversible. The minimal effective dose of either sorbitol or trehalose was approximately 0.2 M, a value similar to the in vivo concentration of sorbitol in diapausing eggs (0.2 M). Glycerol, mannitol or glucose were moderately effective for inhibition. Sorbitol present in diapausing silkworm eggs does not appear to serve as an antifreeze, but as an strong arresting factor of embryonic development. Furthermore, these results show that a decrease in sorbitol releases the embryos from diapause at the termination of diapause.

  6. Overexpression and functional characterization of an Aspergillus niger phytase in the fat body of transgenic silkworm, Bombyx mori.

    PubMed

    Xu, Hanfu; Liu, Yaowen; Wang, Feng; Yuan, Lin; Wang, Yuancheng; Ma, Sanyuan; Beneš, Helen; Xia, QingYou

    2014-08-01

    In a previous study, we isolated 1,119 bp of upstream promoter sequence from Bmlp3, a gene encoding a member of the silkworm 30 K storage protein family, and demonstrated that it was sufficient to direct fat body-specific expression of a reporter gene in a transgenic silkworm, thus highlighting the potential use of this promoter for both functional genomics research and biotechnology applications. To test whether the Bmlp3 promoter can be used to produce recombinant proteins in the fat body of silkworm pupae, we generated a transgenic line of Bombyx mori which harbors a codon-optimized Aspergillus niger phytase gene (phyA) under the control of the Bmlp3 promoter. Here we show that the Bmlp3 promoter drives high levels of phyA expression in the fat body, and that the recombinant phyA protein is highly active (99.05 and 54.80 U/g in fat body extracts and fresh pupa, respectively). We also show that the recombinant phyA has two optimum pH ranges (1.5-2.0 and 5.5-6.0), and two optimum temperatures (55 and 37 °C). The activity of recombinant phyA was lost after high-temperature drying, but treating with boiling water was less harmful, its residual activity was approximately 84% of the level observed in untreated samples. These results offer an opportunity not only for better utilization of large amounts of silkworm pupae generated during silk production, but also provide a novel method for mass production of low-cost recombinant phytase using transgenic silkworms.

  7. The oak wilt enigma: perspectives from the Texas epidemic.

    PubMed

    Appel, D N

    1995-01-01

    Ceratocystis fagacearum (Bretz) Hunt, the oak wilt pathogen, is currently causing massivel osses of semievergreen live oaks (Quercus fusiformis Small and Q. virginiana Mill.) in central Texas. Given the relatively limited oak mortality caused by C. fagacearum in the deciduous forests of the North Central, Midwestern, and Mid-Atlantic United States, this Texas epidemic was not anticipated. The intensity of oak wilt in Texas is attributed to a number of factors related to host characteristics and the ability of the pathogen to adapt to limiting environmental conditions. Oak wilt management in semievergreen oaks requires considerable revision of the control techniques previously designed for deciduous oaks. The Texas oak wilt epidemic provides a new perspective from which to evaluate questions concerning oak wilt, including the origins of the pathogen as well as the potential for future losses in unaffected oak forests.

  8. Poison oak rash on the arm (image)

    MedlinePlus

    ... appearance of poison oak dermatitis. Note the typical linear streaks produced either by scratching or brushing against the plant. (Image courtesy of the Centers for Disease Control and Prevention.)

  9. Oak Ridge Reservation environmental report for 1989

    SciTech Connect

    Jacobs, V.A.; Wilson, A.R.

    1990-10-01

    This two-volume report, the Oak Ridge Reservation Environmental Report for 1989, is the nineteenth in an annual series that began in 1971. It reports the results of a comprehensive, year-round program to monitor the impact of operations at the three major US Department of Energy (DOE) production and research installations in Oak Ridge on the immediate areas' and surrounding region's groundwater and surface waters, soil, air quality, vegetation and wildlife, and through these multiple and varied pathways, the resident human population. Information is presented for the environmental monitoring Quality Assurance (QA) Program, audits and reviews, waste management activities, land special environmental studies. Data are included for the Oak Ridge Y-12 Plant, Oak Ridge National Laboratory (ORNL), and Oak Ridge Gaseous Diffusion Plant (ORGDP). Volume 1 presents narratives, summaries, and conclusions based on environmental monitoring at the three DOE installations and in the surrounding environs during calendar year (CY) 1989. Volume 1 is intended to be a stand-alone'' report about the Oak Ridge Reservation (ORR) for the reader who does not want an in-depth review of 1989 data. Volume 2 presents the detailed data from which these conclusions have been drawn and should be used in conjunction with Volume 1.

  10. The development of an aquatic spill model for the White Oak Creek watershed, Oak Ridge National Laboratory, Oak Ridge, Tennessee

    SciTech Connect

    Johnson, R.O.

    1996-05-01

    This study develops an aquatic spill model applicable to the White Oak Creek watershed draining the Oak Ridge National Laboratory. Hazardous, toxic, and radioactive chemicals are handled and stored on the laboratory reservation. An accidental spill into the White Oak Creek watershed could contaminate downstream water supplies if insufficient dilution did not occur. White Oak Creek empties into the Clinch River, which flows into the Tennessee River. Both rivers serve as municipal water supplies. The aquatic spill model provides estimates of the dilution at sequential downstream locations along White Oak creek and the Clinch River after an accidental spill of a liquid containing a radioactively decaying constituent. The location of the spill on the laboratory is arbitrary, while hydrologic conditions range from drought to extreme flood are simulated. The aquatic spill model provides quantitative estimates with which to assess water quality downstream from the site of the accidental spill, allowing an informed decision to be made whether to perform mitigating measures so that the integrity of affected water supplies is not jeopardized.

  11. Embryonic thermosensitive TRPA1 determines transgenerational diapause phenotype of the silkworm, Bombyx mori.

    PubMed

    Sato, Azusa; Sokabe, Takaaki; Kashio, Makiko; Yasukochi, Yuji; Tominaga, Makoto; Shiomi, Kunihiro

    2014-04-01

    In the bivoltine strain of the silkworm, Bombyx mori, embryonic diapause is induced transgenerationally as a maternal effect. Progeny diapause is determined by the environmental temperature during embryonic development of the mother; however, its molecular mechanisms are largely unknown. Here, we show that the Bombyx TRPA1 ortholog (BmTrpA1) acts as a thermosensitive transient receptor potential (TRP) channel that is activated at temperatures above ∼ 21 °C and affects the induction of diapause in progeny. In addition, we show that embryonic RNAi of BmTrpA1 affects diapause hormone release during pupal-adult development. This study identifying a thermosensitive TRP channel that acts as a molecular switch for a relatively long-term predictive adaptive response by inducing an alternative phenotype to seasonal polyphenism is unique.

  12. A novel sodium N-fatty acyl amino acid surfactant using silkworm pupae as stock material.

    PubMed

    Wu, Min-Hui; Wan, Liang-Ze; Zhang, Yu-Qing

    2014-01-01

    A novel sodium N-fatty acyl amino acid (SFAAA) surfactant was synthesized using pupa oil and pupa protein hydrolysates (PPH) from a waste product of the silk industry. The aliphatic acids from pupa oil were modified into N-fatty acyl chlorides by thionyl chloride (SOCl2). SFAAA was synthesized using acyl chlorides and PPH. GC-MS analysis showed fatty acids from pupa oil consist mainly of unsaturated linolenic and linoleic acids and saturated palmitic and stearic acids. SFAAA had a low critical micelle concentration, great efficiency in lowering surface tension and strong adsorption at an air/water interface. SFAAA had a high emulsifying power, as well as a high foaming power. The emulsifying power of PPH and SFAAA in an oil/water emulsion was better with ethyl acetate as the oil phase compared to n-hexane. The environment-friendly surfactant made entirely from silkworm pupae could promote sustainable development of the silk industry. PMID:24651079

  13. A novel sodium N-fatty acyl amino acid surfactant using silkworm pupae as stock material.

    PubMed

    Wu, Min-Hui; Wan, Liang-Ze; Zhang, Yu-Qing

    2014-03-21

    A novel sodium N-fatty acyl amino acid (SFAAA) surfactant was synthesized using pupa oil and pupa protein hydrolysates (PPH) from a waste product of the silk industry. The aliphatic acids from pupa oil were modified into N-fatty acyl chlorides by thionyl chloride (SOCl2). SFAAA was synthesized using acyl chlorides and PPH. GC-MS analysis showed fatty acids from pupa oil consist mainly of unsaturated linolenic and linoleic acids and saturated palmitic and stearic acids. SFAAA had a low critical micelle concentration, great efficiency in lowering surface tension and strong adsorption at an air/water interface. SFAAA had a high emulsifying power, as well as a high foaming power. The emulsifying power of PPH and SFAAA in an oil/water emulsion was better with ethyl acetate as the oil phase compared to n-hexane. The environment-friendly surfactant made entirely from silkworm pupae could promote sustainable development of the silk industry.

  14. A novel sodium N-fatty acyl amino acid surfactant using silkworm pupae as stock material

    NASA Astrophysics Data System (ADS)

    Wu, Min-Hui; Wan, Liang-Ze; Zhang, Yu-Qing

    2014-03-01

    A novel sodium N-fatty acyl amino acid (SFAAA) surfactant was synthesized using pupa oil and pupa protein hydrolysates (PPH) from a waste product of the silk industry. The aliphatic acids from pupa oil were modified into N-fatty acyl chlorides by thionyl chloride (SOCl2). SFAAA was synthesized using acyl chlorides and PPH. GC-MS analysis showed fatty acids from pupa oil consist mainly of unsaturated linolenic and linoleic acids and saturated palmitic and stearic acids. SFAAA had a low critical micelle concentration, great efficiency in lowering surface tension and strong adsorption at an air/water interface. SFAAA had a high emulsifying power, as well as a high foaming power. The emulsifying power of PPH and SFAAA in an oil/water emulsion was better with ethyl acetate as the oil phase compared to n-hexane. The environment-friendly surfactant made entirely from silkworm pupae could promote sustainable development of the silk industry.

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

    PubMed

    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. PMID:23569267

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

    PubMed

    Cenis, Jose Luis; Aznar-Cervantes, Salvador D; Lozano-Pérez, Antonio Abel; Rojo, Marta; Muñoz, Juan; Meseguer-Olmo, Luis; Arenas, Aurelio

    2016-07-16

    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.

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

    PubMed Central

    Cenis, Jose Luis; Aznar-Cervantes, Salvador D.; Lozano-Pérez, Antonio Abel; Rojo, Marta; Muñoz, Juan; Meseguer-Olmo, Luis; Arenas, Aurelio

    2016-01-01

    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. PMID:27438824

  18. Biological Activity of Recombinant Bovine Interferon τ Produced by a Silkworm-Baculovirus Gene Expression System

    PubMed Central

    TAKAHASHI, Hitomi; TSUNAZAKI, Makoto; HAMANO, Takashi; TAKAHASHI, Masashi; OKUDA, Kiyoshi; INUMARU, Shigeki; OKANO, Akira; GESHI, Masaya; HIRAKO, Makoto

    2013-01-01

    ABSTRACT Bovine interferon (bIFN) τ plays a crucial role in maternal-fetal recognition and was expressed using a Bombyx mori (Bm) nuclear polyhedrosis virus (silkworm baculovirus) gene expression system. The biological effects of Bm-recombinant bIFNτ (rbIFNτ) on prostaglandin (PG) F2α synthesis were investigated in cultured bovine endometrial epithelial cells with oxytocin (OT, 100 nM) and on the in vitro development of bovine embryos. Bm-rbIFNτ and OT were shown to suppress PGF2α production in a dose-dependent manner. When in vitro produced morula stage embryos were cultured for 72 hr in modified CR1aa medium supplemented with or without rbIFNτ, Bm-rbIFNτ (10 ng/ml) significantly promoted development to the expanded blastocyst stage. In conclusion, Bm-rbIFNτ was suggested to have the same bioactivity as native IFNτ. PMID:24212505

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

    PubMed

    Cenis, Jose Luis; Aznar-Cervantes, Salvador D; Lozano-Pérez, Antonio Abel; Rojo, Marta; Muñoz, Juan; Meseguer-Olmo, Luis; Arenas, Aurelio

    2016-01-01

    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. PMID:27438824

  20. Transgenic characterization of two testis-specific promoters in the silkworm, Bombyx mori.

    PubMed

    Xu, J; Bi, H; Chen, R; Aslam, A F M; Li, Z; Ling, L; Zeng, B; Huang, Y; Tan, A

    2015-04-01

    Sex-specific regulatory elements are key components for developing insect genetic sexing systems. The current insect genetic sexing system mainly uses a female-specific modification system whereas little success was reported on male-specific genetic modification. In the silkworm Bombyx mori, a lepidopteran model insect with economic importance, a transgene-based, female-specific lethality system has been established based on sex-specific alternative splicing factors and a female-specific promoter BmVgp (vitellogenin promoter) has been identified. However, no male-specific regulatory elements have yet been identified. Here we report the transgenic identification of two promoters that drive reporter gene expression in a testis-specific manner in B. mori. Putative promoter sequences from the B. mori Radial spoke head 1 gene (BmR1) and beta-tubulin 4 gene (Bmβ4) were introduced using piggybac-based germline transformation. In transgenic silkworms, expression of the reporter gene enhanced green fluorescent protein (EGFP) directed by either BmR1 promoter (BmR1p) or Bmβ4p showed precisely testis-specific manners from the larval to adult stage. Furthermore, EGFP expression of these two transgenic lines showed different localization in the testis, indicating that BmR1p or Bmβ4p might be used as distinct regulatory elements in directing testis-specific gene expression. Identification of these testis-specific promoters not only contributes to a better understanding of testis-specific gene function in insects, but also has potential applications in sterile insect techniques for pest management.

  1. Promoter analysis and RNA interference of CYP6ab4 in the silkworm Bombyx mori.

    PubMed

    Zhao, Guo-Dong; Zhang, Yi-Ling; Liu, Yun-Lei; Li, Bing; Chen, Yu-Hua; Xu, Ya-Xiang; Xia, Qing-You; Shen, Wei-De; Wei, Zheng-Guo

    2015-10-01

    In insects, cytochrome P450 monooxygenases (P450s) are involved in the metabolism of endogenous compounds such as steroid hormones and lipids. In this study, we measured the 20-hydroxyecdysone (20E)-induced transcriptional level of the CYP6ab4 gene using reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) with a dual spike-in strategy. We then probed possible physiological functions using RNAi experiments in the silkworm Bombyx mori. The activity of the CYP6ab4 promoter in various silkworm tissues was measured by firefly luciferase activity and normalized by Renilla luciferase activity. Our results showed that the activity of the CYP6ab4 promoter was highest in the malpighian tubule, followed by the fat body, the silk gland, the midgut, the epidermis, and the hemocyte. The essential region for basal and 20E-induced transcriptional activity was between -908 and -456 bp from the transcription start site. Through promoter truncation analysis using a dual-luciferase reporter assay in B. mori ovary cells (BmN), we showed that the region between -827 and -722 bp was essential for basal and 20E-induced transcriptional activity. Sequence analysis of this region revealed several potential transcriptional regulatory elements such as Hunchback (Hb) and BR-C Z. Mutation of the core bases of the BR-C Z binding site demonstrated that BR-C Z induces 20E-mediated CYP6ab4 transcription. Further identification of cis- and trans-elements and their roles in the upregulation of CYP6ab4 may be useful for elucidating the contribution of P450 to the response mechanism to 20E.

  2. TRANSCRIPTION FACTOR Bmsage PLAYS A CRUCIAL ROLE IN SILK GLAND GENERATION IN SILKWORM, Bombyx mori.

    PubMed

    Xin, Hu-hu; Zhang, Deng-pan; Chen, Rui-ting; Cai, Zi-zheng; Lu, Yan; Liang, Shuang; Miao, Yun-gen

    2015-10-01

    Salivary gland secretion is altered in Drosophila embryos with loss of function of the sage gene. Saliva has a reduced volume and an increased electron density according to transmission electron microscopy, resulting in regions of tube dilation and constriction with intermittent tube closure. However, the precise functions of Bmsage in silkworm (Bombyx mori) are unknown, although its sequence had been deposited in SilkDB. From this, Bmsage is inferred to be a transcription factor that regulates the synthesis of silk fibroin and interacts with another silk gland-specific transcription factor, namely, silk gland factor-1. In this study, we introduced a germline mutation of Bmsage using the Cas9/sgRNA system, a genome-editing technology, resulting in deletion of Bmsage from the genome of B. mori. Of the 15 tested samples, seven displayed alterations at the target site. The mutagenesis efficiency was about 46.7% and there were no obvious off-target effects. In the screened homozygous mutants, silk glands developed poorly and the middle and posterior silk glands (MSG and PSG) were absent, which was significantly different from the wild type. The offspring of G0 mosaic silkworms had indel mutations causing 2- or 9-bp deletions at the target site, but exhibited the same abnormal silk gland structure. Mutant larvae containing different open-reading frames of Bmsage had the same silk gland phenotype. This illustrated that the mutant phenotype was due to Bmsage knockout. We conclude that Bmsage participates in embryonic development of the silk gland. PMID:25917878

  3. Getting a full dose? Reconsidering sex chromosome dosage compensation in the silkworm, Bombyx mori.

    PubMed

    Walters, James R; Hardcastle, Thomas J

    2011-01-01

    Dosage compensation--equalizing gene expression levels in response to differences in gene dose or copy number--is classically considered to play a critical role in the evolution of heteromorphic sex chromosomes. As the X and Y diverge through degradation and gene loss on the Y (or the W in female-heterogametic ZW taxa), it is expected that dosage compensation will evolve to correct for sex-specific differences in gene dose. Although this is observed in some organisms, recent genome-wide expression studies in other taxa have revealed striking exceptions. In particular, reports that both birds and the silkworm moth (Bombyx mori) lack dosage compensation have spurred speculation that this is the rule for all female-heterogametic taxa. Here, we revisit the issue of dosage compensation in silkworm by replicating and extending the previous analysis. Contrary to previous reports, our efforts reveal a pattern typically associated with dosage compensated taxa: the global male:female expression ratio does not differ between the Z and autosomes. We believe the previous report of unequal male:female ratios on the Z reflects artifacts of microarray normalization in conjunction with not testing a major assumption that the male:female global expression ratio was unbiased for autosomal loci. However, we also find that the global Z chromosome expression is significantly reduced relative to autosomes, a pattern not expected in dosage compensated taxa. This combination of male:female parity with an overall reduction in expression for sex-linked loci is not consistent with the prevailing evolutionary theory of sex chromosome evolution and dosage compensation.

  4. Transgenic characterization of two testis-specific promoters in the silkworm, Bombyx mori.

    PubMed

    Xu, J; Bi, H; Chen, R; Aslam, A F M; Li, Z; Ling, L; Zeng, B; Huang, Y; Tan, A

    2015-04-01

    Sex-specific regulatory elements are key components for developing insect genetic sexing systems. The current insect genetic sexing system mainly uses a female-specific modification system whereas little success was reported on male-specific genetic modification. In the silkworm Bombyx mori, a lepidopteran model insect with economic importance, a transgene-based, female-specific lethality system has been established based on sex-specific alternative splicing factors and a female-specific promoter BmVgp (vitellogenin promoter) has been identified. However, no male-specific regulatory elements have yet been identified. Here we report the transgenic identification of two promoters that drive reporter gene expression in a testis-specific manner in B. mori. Putative promoter sequences from the B. mori Radial spoke head 1 gene (BmR1) and beta-tubulin 4 gene (Bmβ4) were introduced using piggybac-based germline transformation. In transgenic silkworms, expression of the reporter gene enhanced green fluorescent protein (EGFP) directed by either BmR1 promoter (BmR1p) or Bmβ4p showed precisely testis-specific manners from the larval to adult stage. Furthermore, EGFP expression of these two transgenic lines showed different localization in the testis, indicating that BmR1p or Bmβ4p might be used as distinct regulatory elements in directing testis-specific gene expression. Identification of these testis-specific promoters not only contributes to a better understanding of testis-specific gene function in insects, but also has potential applications in sterile insect techniques for pest management. PMID:25387604

  5. Targeted gene expression using the GAL4/UAS system in the silkworm Bombyx mori.

    PubMed Central

    Imamura, Morikazu; Nakai, Junichi; Inoue, Satoshi; Quan, Guo Xing; Kanda, Toshio; Tamura, Toshiki

    2003-01-01

    The silkworm Bombyx mori is one of the most well-studied insects in terms of both genetics and physiology and is recognized as the model lepidopteran insect. To develop an efficient system for analyzing gene function in the silkworm, we investigated the feasibility of using the GAL4/UAS system in conjunction with piggyBac vector-mediated germ-line transformation for targeted gene expression. To drive the GAL4 gene, we used two endogenous promoters that originated from the B. mori actin A3 (BmA3) and fibroin light-chain (FiL) genes and the artificial promoter 3xP3. GFP was used as the reporter. In initial tests of the function of the GAL4/UAS system, we generated transgenic animals that carried the UAS-GFP construct plus either BmA3-GAL4 or 3xP3-GAL4. GFP fluorescence was observed in the tissues of GFP-positive animals, in which both promoters drove GAL4 gene expression. Animals that possessed only the GAL4 gene or UAS-GFP construct did not show GFP fluorescence. In addition, as a further test of the ability of the GAL4/UAS system to drive tissue-specific expression we constructed FiL-GAL4 lines with 3xP3-CFP as the transformation marker. FiL-GAL4 x UAS-GFP crosses showed GFP expression in the posterior silk gland, in which the endogenous FiL gene is normally expressed. These results show that the GAL4/UAS system is applicable to B. mori and emphasize the potential of this system for controlled analyses of B. mori gene function. PMID:14668386

  6. Working and Learning Among California Oaks

    NASA Astrophysics Data System (ADS)

    Tietje, B.; Gingg, B.; Zingo, J.; Huntsinger, L.

    2009-04-01

    With tremendous support from collaborators and enthusiastic volunteers, "Learning Among the Oaks" at the historic Santa Margarita Ranch has become a favorite outdoor learning experience for hundreds of Santa Margarita School students, along with their teachers and families. Oaks are at the center of this unique and cost effective public education program. From getting to know local oaks to exploring conservation issues within the context of a historic working cattle ranch, students take pride in expanding their awareness and knowledge of the local oak woodland community. Santa Margarita School families representing the varied demographics of the community come together on the trail. For many, the program provides a first opportunity to get to know those who make a living on the land and to understand that this land around their school is more than a pretty view. "Learning Among the Oaks" also addresses the need for quality, hands-on science activities and opportunities to connect children with the outdoor world. Using a thematic approach and correlating lessons with State Science Standards, we've engaged students in a full-spectrum of exciting outdoor learning adventures. As students progress through the grades, they find new challenges within the oak trail environment. We've succeeded in establishing an internship program that brings highly qualified, enthusiastic university students out to practice their science teaching skills while working with elementary school students. In the future, these university student interns may assist with the development of interpretive displays, after-school nature activities and monitoring projects. We've benefited from proximity to Cal Poly State University and its "learn-by-doing" philosophy. We've also succeeded in building a dedicated network of volunteers and collaborators, each with a special interest satisfied through participation in the oak trail program. While "Learning Among the Oaks" has focused on educating school

  7. ARTICLES: Synthesis of Biomorphic ZrO2-CeO2 Nanostructures by Silkworm Silk Template

    NASA Astrophysics Data System (ADS)

    Zhang, Zong-jian; Li, Jia; Sun, Fu-sheng; Dickon, H. L. Ng; Luen Kwong, Fung

    2010-06-01

    A simple and green technique has been developed to prepare hierarchical biomorphic ZrO2-CeO2, using silkworm silk as the template. Different from traditional immersion technics, the whole synthesis process depends more on the restriction or direction functions of the silkworm silk template. The analytic results showed that ZrO2-CeO2 exhibited a well-crystallized hierarchically interwoven hollow fiber structure with 16-28 μm in diameter. The grain size of the sample calcined at 800 °C was about 14 nm. Consequently, the interwoven meshwork at three dimensions is formed due to the direction of biotemplate. The action mechanism is summarily discussed here. It may bring the biomorphic ZrO2-CeO2 nanomaterials with hierarchical interwoven structures to more applications, such as catalysts.

  8. Cloning and expression analysis of the Bombyx mori α-amylase gene (Amy) from the indigenous Thai silkworm strain, Nanglai.

    PubMed

    Ngernyuang, Nipaporn; Kobayashi, Isao; Promboon, Amornrat; Ratanapo, Sunanta; Tamura, Toshiki; Ngernsiri, Lertluk

    2011-01-01

    α-Amylase is a common enzyme for hydrolyzing starch. In the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), α-amylase is found in both digestive fluid and hemolymph. Here, the complete genomic sequence of the Amy gene encoding α-amylase from a local Thai silkworm, the Nanglai strain, was obtained. This gene was 7981 bp long with 9 exons. The full length Amy cDNA sequence was 1749 bp containing a 1503 bp open reading frame. The ORF encoded 500 amino acid residues. The deduced protein showed 81-54% identity to other insect α-amylases and more than 50% identity to mammalian enzymes. Southern blot analysis revealed that in the Nanglai strain Amy is a single-copy gene. RT- PCR showed that Amy was transcribed only in the foregut. Transgenic B. mori also showed that the Amy promoter activates expression of the transgene only in the foregut.

  9. The heuristic function of 'error' in the scientific methodology of Louis Pasteur: the case of the silkworm diseases.

    PubMed

    Cadeddu, A

    2000-01-01

    With the aid of the Cahiers de laboratoire, the Correspondence and, of course, the Oeuvre de Pasteur, this work reconstructs the extraordinary scientific undertakings of the great French scientist in his study of silkworm diseases. The focus of this study consists in the attempt to explain the initial perplexing behaviour of Pasteur, even in the presence of correct interpretations regarding the causes of these diseases (cfr. the results obtained by Béchamp); for a good three years he insisted on maintaining that the aetiology of silkworm diseases could not be attributed to pathogenetic germs from outside. And this was in spite of the fact that previously (through fermentation and spontaneous generation) he had been able to demonstrate the importance of microorganisms in biological processes. Finally it is intended to highlight the extraordinary methodological depth of that initial 'error', which was capable of paving the way for the future conquests of Pasteur in the field of aetiology and the prevention of infectious diseases.

  10. Insect food for astronauts: gas exchange in silkworms fed on mulberry and lettuce and the nutritional value of these insects for human consumption during deep space flights.

    PubMed

    Tong, L; Yu, X; Liu, H

    2011-10-01

    In this study, silkworm moth (Bombyx mori L.) larvae were regarded as an animal protein source for astronauts in the bioregenerative life support system during long-term deep space exploration in the future. They were fed with mulberry and stem lettuce leaves during the first three instars and the last two instars, respectively. In addition, this kind of environmental approach, which utilised inedible biomass of plants to produce animal protein of high quality, can likewise be applied terrestrially to provide food for people living in extreme environments and/or impoverished agro-ecosystems, such as in polar regions, isolated military bases, ships, submarines, etc. Respiration characteristics of the larvae during development under two main physiological conditions, namely eating and not-eating of leaves, were studied. Nutrient compositions of silkworm powder (SP), ground and freeze-dried silkworms on the 3rd day of the 5th instar larvae, including protein, fat, vitamins, minerals and fatty acids, were measured using international standard methods. Silkworms' respiration rates, measured when larvae were eating mulberry leaves, were higher than those of similar larvae that hadn't eaten such leaves. There was a significant difference between silkworms fed on mulberry leaves and those fed on stem lettuce in the 4th and 5th instars (P<0.01). Amounts of CO2 exhaled by the silkworms under the two physiological regimes differed from each other (P<0.01). There was also a significant difference between the amount of O2 inhaled when the insects were under the two physiological statuses (P<0.01). Moreover, silkworms' respiration quotient under the eating regime was larger than when under the not-eating regime. The SP was found to be rich in protein and amino acids in total; 12 essential vitamins, nine minerals and twelve fatty acids were detected. Moreover, 359 kcal could be generated per 100 gram of SP (dry weight).

  11. Screening of contaminants in Waste Area Grouping 2 at Oak Ridge National Laboratory, Oak Ridge, Tennessee

    SciTech Connect

    Blaylock, B.G.; Frank, M.L.; Hoffman, F.O.; Hook, L.A.; Suter, G.W.; Watts, J.A.

    1992-07-01

    Waste Area Grouping 2 (WAG 2) of the Oak Ridge National Laboratory (ORNL) is located in the White Oak Creek Watershed and is composed of White Oak Creek Embayment, White Oak Lake and associated floodplain, and portions of White Oak Creek (WOC) and Melton Branch downstream of ORNL facilities. Contaminants leaving other ORNL WAGs in the WOC watershed pass through WAG 2 before entering the Clinch River. Health and ecological risk screening analyses were conducted on contaminants in WAG 2 to determine which contaminants were of concern and would require immediate consideration for remedial action and which contaminants could be assigned a low priority or further study. For screening purposes, WAG 2 was divided into four geographic reaches: Reach 1, a portion of WOC; Reach 2, Melton Branch; Reach 3, White Oak Lake and the floodplain area to the weirs on WOC and Melton Branch; and Reach 4, the White Oak Creek Embayment, for which an independent screening analysis has been completed. Screening analyses were conducted using data bases compiled from existing data on carcinogenic and noncarcinogenic contaminants, which included organics, inorganics, and radionuclides. Contaminants for which at least one ample had a concentration above the level of detection were placed in a detectable contaminants data base. Those contaminants for which all samples were below the level of detection were placed in a nondetectable contaminants data base.

  12. Development of oak plantations established for wildlife

    USGS Publications Warehouse

    Twedt, D.J.; Wilson, R.R.

    2002-01-01

    Extensive areas that are currently in agricultural production within the Mississippi Alluvial Valley are being restored to bottomland hardwood forests. Oaks (Quercus sp.), sown as seeds (acorns) or planted as seedlings, are the predominant trees established on most afforested sites. To compare stand development and natural invasion on sites afforested by planting seedlings or by sowing acorns, we sampled woody vegetation on ten 14- to 18-year-old oak plantations established to provide wildlife habitat. Stem densities of about 900 oaks/ha were comparable between stands established by sowing 4000 acorns/ha and stands established by planting 900 seedlings/ha. Densities of oaks in stands established from seedlings increased 38% from densities detected when these stands were 4- to 8-year-old. Densities of oaks established from field-sown acorns increased >100% during this same 10-year span. Oaks that were planted as seedlings were larger than those established from acorns, but trees resulting from either afforestation method were larger than trees naturally colonizing these sites. Natural invasion of woody species varied greatly among afforested sites, but was greater and more diverse on sites sown with acorns. Afforested stands were dominated by planted species, whereas naturally invading species were rare among dominant canopy trees. When afforestation objectives are primarily to provide wildlife habitat, we recommend, sowing acorns rather than planting seedlings. Additionally, planting fewer seeds or seedlings, diversifying the species planted, and leaving non-planted gaps will increase diversity of woody species and promote a more complex forest structure that enhances the suitability of afforested sites for wildlife.

  13. Comparative analysis of the integument transcriptomes of the black dilute mutant and the wild-type silkworm Bombyx mori

    PubMed Central

    Wu, Songyuan; Tong, Xiaoling; Peng, Chenxing; Xiong, Gao; Lu, Kunpeng; hu, Hai; Tan, Duan; Li, Chunlin; Han, Minjin; Lu, Cheng; Dai, Fangyin

    2016-01-01

    The insect cuticle is a critical protective shell that is composed predominantly of chitin and various cuticular proteins and pigments. Indeed, insects often change their surface pigment patterns in response to selective pressures, such as threats from predators, sexual selection and environmental changes. However, the molecular mechanisms underlying the construction of the epidermis and its pigmentation patterns are not fully understood. Among Lepidoptera, the silkworm is a favorable model for color pattern research. The black dilute (bd) mutant of silkworm is the result of a spontaneous mutation; the larval body color is notably melanized. We performed integument transcriptome sequencing of the wild-type strain Dazao and the mutant strains +/bd and bd/bd. In these experiments, during an early stage of the fourth molt, a stage at which approximately 51% of genes were expressed genome wide (RPKM ≥1) in each strain. A total of 254 novel transcripts were characterized using Cuffcompare and BLAST analyses. Comparison of the transcriptome data revealed 28 differentially expressed genes (DEGs) that may contribute to bd larval melanism, including 15 cuticular protein genes that were remarkably highly expressed in the bd/bd mutant. We suggest that these significantly up-regulated cuticular proteins may promote melanism in silkworm larvae. PMID:27193628

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

    PubMed

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

    2016-05-01

    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. PMID:26972338

  15. Quantitative proteomic and transcriptomic analyses of molecular mechanisms associated with low silk production in silkworm Bombyx mori.

    PubMed

    Wang, Shao-Hua; You, Zheng-Ying; Ye, Lu-Peng; Che, Jiaqian; Qian, Qiujie; Nanjo, Yohei; Komatsu, Setsuko; Zhong, Bo-Xiong

    2014-02-01

    To investigate the molecular mechanisms underlying the low fibroin production of the ZB silkworm strain, we used both SDS-PAGE-based and gel-free-based proteomic techniques and transcriptomic sequencing technique. Combining the data from two different proteomic techniques was preferable in the characterization of the differences between the ZB silkworm strain and the original Lan10 silkworm strain. The correlation analysis showed that the individual protein and transcript were not corresponded well, however, the differentially changed proteins and transcripts showed similar regulated direction in function at the pathway level. In the ZB strain, numerous ribosomal proteins and transcripts were down-regulated, along with the transcripts of translational related elongation factors and genes of important components of fibroin. The proteasome pathway was significantly enhanced in the ZB strain, indicating that protein degradation began on the third day of fifth instar when fibroin would have been produced in the Lan10 strain normally and plentifully. From proteome and transcriptome levels of the ZB strain, the energy-metabolism-related pathways, oxidative phosphorylation, glycolysis/gluconeogenesis, and citrate cycle were enhanced, suggesting that the energy metabolism was vigorous in the ZB strain, while the silk production was low. This may due to the inefficient energy employment in fibroin synthesis in the ZB strain. These results suggest that the reason for the decreasing of the silk production might be related to the decreased ability of fibroin synthesis, the degradation of proteins, and the inefficiency of the energy exploiting.

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

    PubMed

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

    2016-05-01

    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.

  17. Comparative analysis of the integument transcriptomes of the black dilute mutant and the wild-type silkworm Bombyx mori.

    PubMed

    Wu, Songyuan; Tong, Xiaoling; Peng, Chenxing; Xiong, Gao; Lu, Kunpeng; Hu, Hai; Tan, Duan; Li, Chunlin; Han, Minjin; Lu, Cheng; Dai, Fangyin

    2016-01-01

    The insect cuticle is a critical protective shell that is composed predominantly of chitin and various cuticular proteins and pigments. Indeed, insects often change their surface pigment patterns in response to selective pressures, such as threats from predators, sexual selection and environmental changes. However, the molecular mechanisms underlying the construction of the epidermis and its pigmentation patterns are not fully understood. Among Lepidoptera, the silkworm is a favorable model for color pattern research. The black dilute (bd) mutant of silkworm is the result of a spontaneous mutation; the larval body color is notably melanized. We performed integument transcriptome sequencing of the wild-type strain Dazao and the mutant strains +/bd and bd/bd. In these experiments, during an early stage of the fourth molt, a stage at which approximately 51% of genes were expressed genome wide (RPKM ≥1) in each strain. A total of 254 novel transcripts were characterized using Cuffcompare and BLAST analyses. Comparison of the transcriptome data revealed 28 differentially expressed genes (DEGs) that may contribute to bd larval melanism, including 15 cuticular protein genes that were remarkably highly expressed in the bd/bd mutant. We suggest that these significantly up-regulated cuticular proteins may promote melanism in silkworm larvae. PMID:27193628

  18. Large-scale production of porcine mature interleukin-18 (IL-18) in silkworms using a hybrid baculovirus expression system.

    PubMed

    Muneta, Yoshihiro; Zhao, Hong Kun; Inumaru, Shigeki; Mori, Yasuyuki

    2003-02-01

    In this report, a hybrid baculovirus expression system, which means a hybrid virus of the Autographa californica nuclear polyhedrosis virus and the Bombyx mori nuclear polyhedrosis virus, was used for the large-scale production of porcine mature interleukin-18 (IL-18) in silkworms. Two recombinant hybrid baculoviruses containing cDNA of the porcine precursor IL-18 and the porcine caspase-1 were constructed and were used to infect silkworm larvae. After the co-infection of the two viruses, porcine mature IL-18 was efficiently produced in the haemolymph. The concentration of IL-18 in the haemolymph was 80-100 microg/ml, as determined by porcine IL-18 specific ELISA. This yield was twenty-times more than that of the insect cell expression system described previously. The porcine mature IL-18 produced by the silkworms strongly induced interferon-gamma (IFN-gamma) production from porcine PBMC. An insect factory system for the large-scale production of useful cytokines for livestock animals will be available in the near future. PMID:12655117

  19. Identification of a microspordium isolated from Megacopta cribraria (Hemiptera: Plataspidae) and characterization of its pathogenicity in silkworms.

    PubMed

    Xing, Dongxu; Li, Li; Liao, Sentai; Luo, Guoqing; Li, Qingrong; Xiao, Yang; Dai, Fanwei; Yang, Qiong

    2014-11-01

    A new microsporidium isolated from Megacopta cribraria was characterized by both biological characteristics and phylogenetic analysis. Moreover, its pathogenicity to silkworms was also studied. The spores are oval in shape and measured 3.64 ± 0.2 × 2.20 ± 0.2 μm in size. Its ultrastructure is characteristic of the genus Nosema: a diplokaryon, 13-14 polar filament coils and posterior vacuole. Its life cycle includes meronts, sporonts, sporoblasts and mature spores, with a typical diplokaryon in each stage and propagation in a binary fission. A phylogenetic tree based on SSU rRNA and rRNA ITS gene sequence analysis further indicated that the parasite is closely related to Nosema bombycis and should be placed in the genus Nosema and sub-group 'true' Nosema. Furthermore, the microsporidium heavily infects lepidopteran silkworm insect and can be transmitted per os (horizontally) and transovarially (vertically). Our findings showed that the microsporidium belongs to the 'true' Nosema group within the genus Nosema and heavily infects silkworms. Based on the information obtained during this study, we named this new microsporidium isolated from M. cribraria as Nosema sp. MC. PMID:25173855

  20. Effects of Cattle Management on Oak Regeneration in Northern Californian Mediterranean Oak Woodlands

    PubMed Central

    López-Sánchez, Aida; Schroeder, John; Roig, Sonia; Sobral, Mar; Dirzo, Rodolfo

    2014-01-01

    Oak woodlands of Mediterranean ecosystems, a major component of biodiversity hotspots in Europe and North America, have undergone significant land-use change in recent centuries, including an increase in grazing intensity due to the widespread presence of cattle. Simultaneously, a decrease in oak regeneration has been observed, suggesting a link between cattle grazing intensity and limited oak regeneration. In this study we examined the effect of cattle grazing on coast live oak (Quercus agrifolia Née) regeneration in San Francisco Bay Area, California. We studied seedling, sapling and adult density of coast live oak as well as vertebrate herbivory at 8 independent sites under two grazing conditions: with cattle and wildlife presence (n = 4) and only with wildlife (n = 4). The specific questions we addressed are: i) to what extent cattle management practices affect oak density, and ii) what is the effect of rangeland management on herbivory and size of young oak plants. In areas with cattle present, we found a 50% reduction in young oak density, and plant size was smaller, suggesting that survival and growth young plants in those areas are significantly limited. In addition, the presence of cattle raised the probability and intensity of herbivory (a 1.5 and 1.8-fold difference, respectively). These results strongly suggest that the presence of cattle significantly reduced the success of young Q. agrifolia through elevated herbivory. Given the potential impact of reduced recruitment on adult populations, modifying rangeland management practices to reduce cattle grazing pressure seems to be an important intervention to maintain Mediterranean oak woodlands. PMID:25126939

  1. View of New Big Oak Flat Road seen from Old ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    View of New Big Oak Flat Road seen from Old Wawona Road near location of photograph HAER CA-148-17. Note road cuts, alignment, and tunnels. Devils Dance Floor at left distance. Looking northwest - Big Oak Flat Road, Between Big Oak Flat Entrance & Merced River, Yosemite Village, Mariposa County, CA

  2. Selection of reference genes for analysis of stress-responsive genes after challenge with viruses and temperature changes in the silkworm Bombyx mori.

    PubMed

    Guo, Huizhen; Jiang, Liang; Xia, Qingyou

    2016-04-01

    Viruses and high temperature (HT) are the primary threats to silkworms. Changes in the expression of stress-response genes can be measured using quantitative polymerase chain reaction (qPCR) after exposure to viruses or HT. However, appropriate reference genes (RGs) for qPCR data normalization have not been established in this organism. In this study, we summarized the RGs used in the previous silkworm studies after infection with Bombyx mori nucleopolyhedrovirus (BmNPV), B. mori cytoplasmic polyhedrosis virus (BmCPV), or B. mori densovirus (BmDNV) or after HT treatment. The expression levels of these RGs were extracted from silkworm transcriptome data to screen for candidate RGs that were unaffected by the experimental conditions. Actin-1 (A1), actin-3 (A3), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation initiation factor 4a (TIF-4A) were selected for further qPCR verification. The results of RNA-seq and qPCR showed that GAPDH and TIF-4A were suitable RGs after BmNPV challenge or HT stress, whereas TIF-4A was an appropriate RG for BmCPV or BmDNV-Z challenge in silkworms. These results suggested that TIF-4A may be the most appropriate RG for gene expression analysis after challenge with viruses or HT in silkworms.

  3. Species-specific expansion of C2H2 zinc-finger genes and their expression profiles in silkworm, Bombyx mori.

    PubMed

    Duan, Jun; Xia, Qingyou; Cheng, Daojun; Zha, Xingfu; Zhao, Ping; Xiang, Zhonghuai

    2008-12-01

    Most C2H2 zinc-finger proteins (ZFPs) function as sequence-specific DNA-binding transcription factors, and play important roles in a variety of biology processes, such as development, differentiation, and tumor suppression. By searching the silkworm genome with a HMM model of C2H2 zinc-fingers, we have identified a total of 338 C2H2 ZFPs. Most of the ZFP genes were clustered on chromosomes and showed uneven distribution in the genome. Over one third of genes were concentrated on chromosome 11, 15 and 24. Phylogenetic analysis classified all silkworm C2H2 ZFPs into 75 families; 63 of which belong to evolutionarily conserved families. In addition, 188 C2H2 ZFP genes (55.6%) are species-specific to the silkworm. A species-specific expansion of a family with 39 members in a tandem array on chromosome 24 may explain the higher number of species-specific ZFPs in silkworm compared to other organisms. The expression patterns of C2H2 ZFP genes were also examined by microarray analysis. Most of these genes were actively expressed among different tissues on day 3 of the fifth instar. The results provide insight into the biological functions of the silkworm C2H2 ZFP genes in metamorphism and development. PMID:18835444

  4. Oak Ridge reservation land-use plan

    SciTech Connect

    Bibb, W. R.; Hardin, T. H.; Hawkins, C. C.; Johnson, W. A.; Peitzsch, F. C.; Scott, T. H.; Theisen, M. R.; Tuck, S. C.

    1980-03-01

    This study establishes a basis for long-range land-use planning to accommodate both present and projected DOE program requirements in Oak Ridge. In addition to technological requirements, this land-use plan incorporates in-depth ecological concepts that recognize multiple uses of land as a viable option. Neither environmental research nor technological operations need to be mutually exclusive in all instances. Unique biological areas, as well as rare and endangered species, need to be protected, and human and environmental health and safety must be maintained. The plan is based on the concept that the primary use of DOE land resources must be to implement the overall DOE mission in Oak Ridge. This document, along with the base map and overlay maps, provides a reasonably detailed description of the DOE Oak Ridge land resources and of the current and potential uses of the land. A description of the land characteristics, including geomorphology, agricultural productivity and soils, water courses, vegetation, and terrestrial and aquatic animal habitats, is presented to serve as a resource document. Essentially all DOE land in the Oak Ridge area is being fully used for ongoing DOE programs or has been set aside as protected areas.

  5. Oak Ridge Reservation Waste Management Plan

    SciTech Connect

    Turner, J.W.

    1995-02-01

    This report presents the waste management plan for the Oak Ridge Reservation facilities. The primary purpose is to convey what facilities are being used to manage wastes, what forces are acting to change current waste management systems, and what plans are in store for the coming fiscal year.

  6. Consort contact dermatitis due to oak moss.

    PubMed

    Held, J L; Ruszkowski, A M; Deleo, V A

    1988-02-01

    An allergic contact dermatitis in a woman was found to be due to oak moss in her husband's after-shave lotion. When routine patch testing reveals a positive reaction, the dermatologist should consider exposure to the antigen not only in the patient but also through contact with the patient's consort.

  7. Oak Park Middle School and General Electric.

    ERIC Educational Resources Information Center

    Buckley, Anita

    1987-01-01

    Describes a partnership between Oak Park Middle School in Decatur, Alabama, and the General Electric Company that features the use of the quality circle approach. Not only were teachers, administrators, and parents organized into school quality circles, but students also formed quality circles to solve problems at the school. (CH)

  8. Oak ecosystem succession of the Northern Caucasus

    NASA Astrophysics Data System (ADS)

    Chalaya, Elena; Slepykh, Victor; Efimenko, Natalia; Povolotskaya, Nina

    2014-05-01

    English oak (Quercus robur L.) along with its well-known good properties has a high sanitary-hygienic and curative potential. Its volatile metabolites (VM) influence bacteriostatically Staphylococcus aureus 209r, oppressing it in vitro by 85% compared with the control, and Escherichia coli by 45%. There is the least amount of epiphytic microorganisms on the leaves of Q. robur L. compared with some tree species [1]. In addition, VM of Q. robur L. have direct milieu (hypotensive) effects on the organism under its canopy, lowering blood pressure by 20-25 mm Hg [2]. A.P. Kazankin (1993) [4] calculated the prehistoric formula of forest species composition of Caucasian Mineral Waters region (Northern Caucasus): 6Qr3Crp1Fr which was based on the theory of calcium-magnesium absorption complex. According to the theory, underground mineral water, soil, forest litter and the leaves of ground vegetation of the area have the same ratio of the cations Ca and Mg - calcium-magnesium index [3]. Hence oldgrowth in the region consisted of oak (Qr) by 60%, hornbeam (Carpinus-Crp) by 30% and ash (Fraxinus-Fr) only by 10%. Currently, the formula of the forests of the region has been changed by man: 5Fr3Crp2Qr. The proportion of oak forests has decreased to 20%, the proportion of ash has increased by 50%, but the proportion of hornbeam hasn't changed. So it is relevant to restore oak forests of the region in the former ratio to other forest-forming species - ash and hornbeam. Taking into consideration the change of economic formation of society in Russia, it is extremely important to restore natural seed oak forests. Therein the luminance of surface areal is a limiting factor. We have calculated that the natural recovery of oak forests is possible providing observation of optimal moisture standards and soil fertility in combination with solar light within 10-24% from the light of open space. Measures for promotion of oak natural regeneration in mountain resorts of the Northern Caucasus

  9. 78 FR 2431 - Notice of Inventory Completion: U.S. Department of Energy Oak Ridge Office, Oak Ridge, TN...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-11

    .... Department of Energy Oak Ridge Office has corrected an inventory of human remains and associated funerary... of Energy Oak Ridge Office. Repatriation of the human remains and associated funerary object to the... object in the possession of the U.S. Department of Energy Oak Ridge Office. The human remains...

  10. 76 FR 80433 - In the Matter of Royal Oak Capital Management, LLC, 6173 Bellevue Road, Royal Oak, MD 21662...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-23

    ... From the Federal Register Online via the Government Publishing Office SECURITIES AND EXCHANGE COMMISSION In the Matter of Royal Oak Capital Management, LLC, 6173 Bellevue Road, Royal Oak, MD 21662...''), cancelling the registration of Royal Oak Capital Management, LLC, hereinafter referred to as the...

  11. GC/MS-based metabolomic studies reveal key roles of glycine in regulating silk synthesis in silkworm, Bombyx mori.

    PubMed

    Chen, Quanmei; Liu, Xinyu; Zhao, Ping; Sun, Yanhui; Zhao, Xinjie; Xiong, Ying; Xu, Guowang; Xia, Qingyou

    2015-02-01

    Metabolic profiling of silkworm, especially the factors that affect silk synthesis at the metabolic level, is little known. Herein, metabolomic method based on gas chromatography-mass spectrometry was applied to identify key metabolic changes in silk synthesis deficient silkworms. Forty-six differential metabolites were identified in Nd group with the defect of silk synthesis. Significant changes in the levels of glycine and uric acid (up-regulation), carbohydrates and free fatty acids (down-regulation) were observed. The further metabolomics of silk synthesis deficient silkworms by decreasing silk proteins synthesis using knocking out fibroin heavy chain gene or extirpating silk glands operation showed that the changes of the metabolites were almost consistent with those of the Nd group. Furthermore, the increased silk yields by supplying more glycine or its related metabolite confirmed that glycine is a key metabolite to regulate silk synthesis. These findings provide important insights into the regulation between metabolic profiling and silk synthesis.

  12. White Oak Creek watershed: Melton Valley area Remedial Investigation report, at the Oak Ridge National Laboratory, Oak Ridge, Tennessee: Volume 2, Appendixes A and B

    SciTech Connect

    1996-11-01

    This document contains Appendixes A ``Source Inventory Information for the Subbasins Evaluated for the White Oak Creek Watershed`` and B ``Human Health Risk Assessment for White Oak Creek / Melton Valley Area`` for the remedial investigation report for the White Oak Creek Watershed and Melton Valley Area. Appendix A identifies the waste types and contaminants for each subbasin in addition to the disposal methods. Appendix B identifies potential human health risks and hazards that may result from contaminants present in the different media within Oak Ridge National Laboratory sites.

  13. The homeodomain transcription factors antennapedia and POU-M2 regulate the transcription of the steroidogenic enzyme gene Phantom in the silkworm.

    PubMed

    Meng, Meng; Cheng, Dao-Jun; Peng, Jian; Qian, Wen-Liang; Li, Jia-Rui; Dai, Dan-Dan; Zhang, Tian-Lei; Xia, Qing-You

    2015-10-01

    The steroid hormone ecdysone, which controls insect molting and metamorphosis, is synthesized in the prothoracic gland (PG), and several steroidogenic enzymes that are expressed specifically in the PG are involved in ecdysteroidogenesis. In this study, we identified new regulators that are involved in the transcriptional control of the silkworm steroidogenic enzyme genes. In silico analysis predicted several potential cis-regulatory elements (CREs) for the homeodomain transcription factors Antennapedia (Antp) and POU-M2 in the proximal promoters of steroidogenic enzyme genes. Antp and POU-M2 are expressed dynamically in the PG during larval development, and their overexpression in silkworm embryo-derived (BmE) cells induced the expression of steroidogenic enzyme genes. Importantly, luciferase reporter analyses, electrophoretic mobility shift assays, and chromatin immunoprecipitation assays revealed that Antp and POU-M2 promote the transcription of the silkworm steroidogenic enzyme gene Phantom (Phm) by binding directly to specific motifs within overlapping CREs in the Phm promoter. Mutations of these CREs in the Phm promoter suppressed the transcriptional activities of both Antp and POU-M2 in BmE cells and decreased the activities of mutated Phm promoters in the silkworm PG. In addition, pulldown and co-immunoprecipitation assays demonstrated that Antp can interact with POU-M2. Moreover, RNA interference-mediated down-regulation of either Antp or POU-M2 during silkworm wandering not only decreased the ecdysone titer but also led to the failure of metamorphosis. In summary, our results suggest that Antp and POU-M2 coordinate the transcription of the silkworm Phm gene directly, indicating new roles for homeodomain proteins in regulating insect ecdysteroidogenesis.

  14. The Homeodomain Transcription Factors Antennapedia and POU-M2 Regulate the Transcription of the Steroidogenic Enzyme Gene Phantom in the Silkworm*

    PubMed Central

    Meng, Meng; Cheng, Dao-jun; Peng, Jian; Qian, Wen-liang; Li, Jia-rui; Dai, Dan-dan; Zhang, Tian-lei; Xia, Qing-you

    2015-01-01

    The steroid hormone ecdysone, which controls insect molting and metamorphosis, is synthesized in the prothoracic gland (PG), and several steroidogenic enzymes that are expressed specifically in the PG are involved in ecdysteroidogenesis. In this study, we identified new regulators that are involved in the transcriptional control of the silkworm steroidogenic enzyme genes. In silico analysis predicted several potential cis-regulatory elements (CREs) for the homeodomain transcription factors Antennapedia (Antp) and POU-M2 in the proximal promoters of steroidogenic enzyme genes. Antp and POU-M2 are expressed dynamically in the PG during larval development, and their overexpression in silkworm embryo-derived (BmE) cells induced the expression of steroidogenic enzyme genes. Importantly, luciferase reporter analyses, electrophoretic mobility shift assays, and chromatin immunoprecipitation assays revealed that Antp and POU-M2 promote the transcription of the silkworm steroidogenic enzyme gene Phantom (Phm) by binding directly to specific motifs within overlapping CREs in the Phm promoter. Mutations of these CREs in the Phm promoter suppressed the transcriptional activities of both Antp and POU-M2 in BmE cells and decreased the activities of mutated Phm promoters in the silkworm PG. In addition, pulldown and co-immunoprecipitation assays demonstrated that Antp can interact with POU-M2. Moreover, RNA interference-mediated down-regulation of either Antp or POU-M2 during silkworm wandering not only decreased the ecdysone titer but also led to the failure of metamorphosis. In summary, our results suggest that Antp and POU-M2 coordinate the transcription of the silkworm Phm gene directly, indicating new roles for homeodomain proteins in regulating insect ecdysteroidogenesis. PMID:26253172

  15. [Cloning and characterization of BmBrat in silkworm, Bombyx mori].

    PubMed

    Liang, Hanghua; Gao, Hongyan; Xu, Man; Tan, Peng; Cui, Hongjuan

    2016-03-01

    NHL proteins, which play important roles in regulation of cell proliferation and differentiation, have been extensively studied on mammals. Here, we cloned a member of NHL protein family namely BmBrat in silkworm. The full-length cDNA sequence of BmB rat was obtained by means of the rapid amplification of cDNA ends (RACE), including 3 614 bp. The ORF is 2 580 bp long, encoding a protein with 859 amino acid residues. The molecular weight is 94.3 kDa and the isoeledtric point (pI) is 6.65. The BmBrat expression profile was detected by RT-PCR at L5D3 larval stage, and it was expressed in all tissues, including silk gland, midgut, fat body and malpighian tubule. However, it was highly expressed in ovary and head. The expression profile was also detected at different stage of embryo development, and reached a peak at the 4th and 5th days of the embryonic period. Anti-BmBrat polyclonal antibody was generated f6llowing prokaryotic expression, protein purification and mice immunization, which is highly specific and effective for recognizing BmBrat protein through Western blotting and immunofluorescence staining. Subcellular localization of BmBrat in hemocytes revealed that it was specifically expressed in cytoplasm. This study provides a foundation for further research of the biological function of BmBrat gene.

  16. A novel splice variant of the decapentaplegic (dpp) gene in the wild silkworm, Bombyx mandarina.

    PubMed

    Kwak, Woori; Choi, Jung-Won; Ryul Kim, Seong; Choi, Kwang-Ho; Kim, Kee-Young; Goo, Tae-Won; Park, Seung-Won

    2015-10-23

    Decapentaplegic (dpp) is a member of the transforming growth factor-β superfamily. Although the dpp gene and related pathways are known to play important roles in insect development, few studies have examined its function in Bombyx mori and Bombyx mandarina. To date, there have been no previous reports on novel splice variants of dpp in silkworm. In the present study, we conducted RT-PCR to examine dpp expression in the mid-gut tissue of B. mandarina and discovered a novel dpp isoform. The isoform sequence was confirmed using sequencing analysis and found to have 333 bp deletion compared to full-length cDNA encoding dpp. The deleted sequence encodes a region of the latency associated peptide (LAP) region of transforming growth factor-β (TGF-β), which may affect the activity and specificity of TGF-β. Using variant calling analyses, we detected 7 candidate single nucleotide variants (SNVs) for different alternative splicing in dpp. This is the first report of a novel splice variant of the dpp gene in B. mandarina and these results provide insight about the domestication process and distinct phenotypic traits of B. mori and B. mandarina.

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

    NASA Astrophysics Data System (ADS)

    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.

  18. Molecular characterization, tissue distribution, and immune reaction expression of karyopherins in the domestic silkworm (Bombyx mori).

    PubMed

    Li, J; Wang, L; Qian, C; Zhang, C F; Dai, L S; Liu, Q N; Wei, G Q; Sun, Y; Liu, D R; Zhu, B J; Liu, C L

    2015-01-01

    Karyopherins, including alpha and beta types, are transport proteins in the eukaryotic cell that carry cargoes across nuclear pore complexes into or out of the nucleus. In this study, full open reading frames of one beta and three alpha types of karyopherin were cloned from cDNA of the domestic silkworm (Bombyx mori). The one beta and three alpha types' open reading frames were 2661, 1563, 1515, and 1551 base pairs long, respectively, and coded 886, 520, 504, and 516 amino acids, respectively. The alphas all had one importin-beta-binding (IBB) domain, and eight, four, or seven armadillo/beta-catenin-like repeats. The beta had 19 HEAT repeat domains, which constructed one importin-beta-N-terminal domain and one IBB domain. The recombinant proteins were expressed in Escherichia coli cells. The molecular weight of the beta type was approximately 100 kDa, and the alphas weighed approximately 60 kDa. Phylogenic tree construction revealed that the alphas could be classified into three known karyopherin-alpha subfamilies. We detected mRNA of the four karyopherins in normal 3rd day of 5th instar larvae, and in larvae injected with Gram-positive bacteria, Gram-negative bacteria, viruses, and fungi using real-time fluorescence quantitative reverse transcriptase-polymerase chain reaction, and found that the four karyopherins were widely distributed, but their expression levels were related to tissues type, the microbe injected, and the time point.

  19. [Identification and characterization of a novel hemocyte specific gene Bm04862 in silkworm, Bombyx mori].

    PubMed

    Yu, Shuang; Su, Jingjing; Xu, Man; Zhang, Kui; Cui, Hongjuan

    2016-02-01

    Bm04862 is a novel hemocyte-specific gene, which has been identified and cloned through the microarray data in silkworm, Bombyx mori. Initially, we successfully obtained the full-length cDNA sequence of Bm04862 via the rapid amplification of cDNA ends (RACE). The sequence consists of an open reading frame (ORF) with 819 bp, which could encode 273 amino acid residues. The bioinformatics analysis predicted that Bm04862 was a transmembrane protein. Furthermore, qRT-PCR analysis revealed that the expression of Bm04862 was specifically detected in hemocyte and reached a peak at L4M and PP2 stage. Bm04862 was overexpressed in Sf9 insect cells, and the cellular localization indicated that Bm04862 was specifically located in cytoplasm and nuclei membrane. Interestingly, the expression of Bm04862 increas'd dramatically after challenged with Escherichia coli for 24 hours, which predicted its potential role in the innate immune system. Overall, this study could provide the fundamental knowledge for further research.

  20. New insight into the mechanism underlying fibroin secretion in silkworm, Bombyx mori.

    PubMed

    Long, Dingpei; Lu, Weijian; Zhang, Yang; Guo, Qing; Xiang, Zhonghuai; Zhao, Aichun

    2015-01-01

    In order to investigate the role of different parts of the fibroin heavy chain (H-chain) in the secretion of fibroin in the silk gland of the silkworm (Bombyx mori) in vivo, two enhanced green fluorescent protein (EGFP)/H-chain fusion genes with deduced protein sequences containing an identical N-terminal region and different C-terminal regions of the H-chain were introduced into the B. mori genome using a piggyBac-mediated germline transformation. EGFP fluorescence and molecular analysis showed the products of two different EGFP/H-chain fusion proteins were secreted into the posterior silk gland lumen and aggregated in the middle silk gland and spun into cocoons. The results revealed that only the non-repetitive N terminus of the H-chain is essential for secretion of the H-chain into the posterior silk gland lumen. In addition, our results also indicated that the most likely post-translational modification of the H-chain is at the C-terminal domain. Here, our results not only provide a theoretical basis for the genetic modification of silk fiber as a functional biomaterial but also are of great significance to establishing a new silk gland bioreactor to mass-produce exogenous proteins in an active form.

  1. IDENTIFICATION AND EXPRESSION ANALYSIS OF VITELLOGENIN RECEPTOR FROM THE WILD SILKWORM, Bombyx mandarina.

    PubMed

    Qian, Cen; Fu, Wei-Wei; Wei, Guo-Qing; Wang, Lei; Liu, Qiu-Ning; Dai, Li-Shang; Sun, Yu; Zhu, Bao-Jian; Liu, Chao-Liang

    2015-08-01

    The vitellogenin receptor (VgR) plays a key role on embryonic development in oviparous animals. Here, we cloned a VgR gene, which was identified from the wild silkworm Bombyx mandarina (BmaVgR) using reverse transcriptase polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). Sequence analysis revealed that BmaVgR is 5,861 bp long with an open reading frame encoded by 1,811 amino acid residues. The predicted amino acid sequence has 99.7 and 98.2% identity with the VgRs of Actias selene and Bombyx mori, respectively. The class B domain sequence of BmaVgR was cloned and expressed in Escherichia coli, and purified by a Ni-NTA column. Polyclonal antibodies were produced against the purified recombinant protein, and titer of the antibody was about 1:12,800 measured by enzyme-linked immunosorbent assay (ELISA). Western blot and RT-qPCR showed that BmaVgR was expressed in the ovary and fat body of female larvae and the ovary of moth, and the expression level was highest at the third day and then declined from third day to seventh in fat body of pupa. After knockdown of the BmaVgR gene through RNA interference (RNAi), other three BmaVgR-related genes (Vg, egg-specific protein, and low molecular weight lipoprotein LP gene) were all downregulated significantly.

  2. Knockout silkworms reveal a dispensable role for juvenile hormones in holometabolous life cycle

    PubMed Central

    Daimon, Takaaki; Uchibori, Miwa; Nakao, Hajime; Sezutsu, Hideki; Shinoda, Tetsuro

    2015-01-01

    Insect juvenile hormones (JHs) prevent precocious metamorphosis and allow larvae to undergo multiple rounds of status quo molts. However, the roles of JHs during the embryonic and very early larval stages have not been fully understood. We generated and characterized knockout silkworms (Bombyx mori) with null mutations in JH biosynthesis or JH receptor genes using genome-editing tools. We found that embryonic growth and morphogenesis are largely independent of JHs in Bombyx and that, even in the absence of JHs or JH signaling, pupal characters are not formed in first- or second-instar larvae, and precocious metamorphosis is induced after the second instar at the earliest. We also show by mosaic analysis that a pupal specifier gene broad, which is dramatically up-regulated in the late stage of the last larval instar, is essential for pupal commitment in the epidermis. Importantly, the mRNA expression level of broad, which is thought to be repressed by JHs, remained at very low basal levels during the early larval instars of JH-deficient or JH signaling-deficient knockouts. Therefore, our study suggests that the long-accepted paradigm that JHs maintain the juvenile status throughout larval life should be revised because the larval status can be maintained by a JH-independent mechanism in very early larval instars. We propose that the lack of competence for metamorphosis during the early larval stages may result from the absence of an unidentified broad-inducing factor, i.e., a competence factor. PMID:26195792

  3. Positional cloning of a gene responsible for the cts mutation of the silkworm, Bombyx mori.

    PubMed

    Ito, Katsuhiko; Kidokoro, Kurako; Katsuma, Susumu; Shimada, Toru; Yamamoto, Kimiko; Mita, Kazuei; Kadono-Okuda, Keiko

    2012-07-01

    The larval head cuticle and anal plates of the silkworm mutant cheek and tail spot (cts) have chocolate-colored spots, unlike the entirely white appearance of the wild-type (WT) strain. We report the identification and characterization of the gene responsible for the cts mutation. Positional cloning revealed a cts candidate on chromosome 16, designated BmMFS, based on the high similarity of the deduced amino acid sequence between the candidate gene from the WT strain and the major facilitator superfamily (MFS) protein. BmMFS likely encodes a membrane protein with 11 putative transmembrane domains, while the putative structure deduced from the cts-type allele possesses only 10-pass transmembrane domains owing to a deletion in its coding region. Quantitative RT-PCR analysis showed that BmMFS mRNA was strongly expressed in the integument of the head and tail, where the cts phenotype is observed; expression markedly increased at the molting and newly ecdysed stages. These results indicate that the novel BmMFS gene is cts and the membrane structure of its protein accounts for the cts phenotype. These expression profiles and the cts phenotype are quite similar to those of melanin-related genes, such as Bmyellow-e and Bm-iAANAT, suggesting that BmMFS is involved in the melanin synthesis pathway.

  4. Role of GTP-CHI links PAH and TH in melanin synthesis in silkworm, Bombyx mori.

    PubMed

    Chen, Ping; Wang, Jiying; Li, Haiyin; Li, Yan; Chen, Peng; Li, Tian; Chen, Xi; Xiao, Junjie; Zhang, Liang

    2015-08-10

    In insects, pigment patterns are formed by melanin, ommochromes, and pteridines. Here, the effects of pteridine synthesis on melanin formation were studied using 4th instar larvae of a wild-type silkworm strain, dazao (Bombyx mori), with normal color and markings. Results from injected larvae and in vitro integument culture indicated that decreased activity of guanosine triphosphate cyclohydrolase I (GTP-CH I, a rate-limiting enzyme for pteridine synthesis), lowers BH4 (6R-l-erythro-5,6,7,8-tetrahydrobiopterin, a production correlated with GTP-CH I activity) levels and eliminates markings and coloration. The conversion of phenylalanine and tyrosine to melanin was prevented when GTP-CH I was inhibited. When BH4 was added, phenylalanine was converted to tyrosine, and the tyrosine concentration increased. Tyrosine was then converted to melanin to create normal markings and coloration. Decreasing GTP-CH I activity did not affect L-DOPA (3,4-l-dihydroxyphenylalanine). GTP-CH I affected melanin synthesis by generating the BH4 used in two key reaction steps: (1) conversion of phenylalanine to tyrosine by PAH (phenylalanine hydroxylase) and (2) conversion of tyrosine to L-DOPA by TH (tyrosine hydroxylase). Expression profiles of BmGTPCH Ia, BmGTPCH Ib, BmTH, and BmPAH in the integument were consistent with the current findings.

  5. Proteins in the Cocoon of Silkworm Inhibit the Growth of Beauveria bassiana

    PubMed Central

    Zhang, Yan; Li, Youshan; Liu, Huawei; Xia, Qingyou; Zhao, Ping

    2016-01-01

    Silk cocoons are composed of fiber proteins (fibroins) and adhesive glue proteins (sericins), which provide a physical barrier to protect the inside pupa. Moreover, other proteins were identified in the cocoon silk, many of which are immune related proteins. In this study, we extracted proteins from the silkworm cocoon by Tris-HCl buffer (pH7.5), and found that they had a strong inhibitory activity against fungal proteases and they had higher abundance in the outer cocoon layers than in the inner cocoon layers. Moreover, we found that extracted cocoon proteins can inhibit the germination of Beauveria bassiana spores. Consistent with the distribution of protease inhibitors, we found that proteins from the outer cocoon layers showed better inhibitory effects against B. bassiana spores than proteins from the inner layers. Liquid chromatography-tandem mass spectrometry was used to reveal the extracted components in the scaffold silk, the outermost cocoon layer. A total of 129 proteins were identified, 30 of which were annotated as protease inhibitors. Protease inhibitors accounted for 89.1% in abundance among extracted proteins. These protease inhibitors have many intramolecular disulfide bonds to maintain their stable structure, and remained active after being boiled. This study added a new understanding to the antimicrobial function of the cocoon. PMID:27032085

  6. Silkworm Thorn Stem Extract Targets RSK2 and Suppresses Solar UV-Induced Cyclooxygenase-2 Expression.

    PubMed

    Kim, Jong-Eun; Lee, Ki Won

    2015-01-01

    Excessive exposure to solar UV (sUV) is associated with numerous human skin disorders, such as carcinogenesis, skin photoaging and skin inflammation. Silkworm Thorn (Cudraniatricuspidata, SW) is a plant belonging to the Moraceae family and widely present throughout Korea, China, and Japan. Most parts of the tree (including the fruit, leaf, stem, root, and bark) is consumable as a functional food or tea. In this study, we found that SW extract (SWE) inhibited the elevated expression of sUV-induced cyclooxygenase (COX)-2 levels in both HaCaT and JB6 cells. Levels of nuclear factor-κB and activator protein-1, two crucial transcription factors involved in COX-2 expression, were elevated by sUV treatment. Treatment with SWE abolished this activation. SWE also inhibited sUV-induced histone H3 phosphorylation. However, sUV-induced phosphorylation of Akt, c-Jun N-terminal kinase and p38 kinase remained unchanged in the presence of SWE. SWE inhibited RSK2 activity, and pull-down assays using SWE-Sepharose beads revealed that SWE binds directly with RSK2 in an ATP-competitive manner. These results suggest a potential for SWE to be developed as a cosmeceutical material and functional food constituent for the promotion of skin health. PMID:26506342

  7. Molecular characterization, tissue distribution, and immune reaction expression of karyopherins in the domestic silkworm (Bombyx mori).

    PubMed

    Li, J; Wang, L; Qian, C; Zhang, C F; Dai, L S; Liu, Q N; Wei, G Q; Sun, Y; Liu, D R; Zhu, B J; Liu, C L

    2015-01-01

    Karyopherins, including alpha and beta types, are transport proteins in the eukaryotic cell that carry cargoes across nuclear pore complexes into or out of the nucleus. In this study, full open reading frames of one beta and three alpha types of karyopherin were cloned from cDNA of the domestic silkworm (Bombyx mori). The one beta and three alpha types' open reading frames were 2661, 1563, 1515, and 1551 base pairs long, respectively, and coded 886, 520, 504, and 516 amino acids, respectively. The alphas all had one importin-beta-binding (IBB) domain, and eight, four, or seven armadillo/beta-catenin-like repeats. The beta had 19 HEAT repeat domains, which constructed one importin-beta-N-terminal domain and one IBB domain. The recombinant proteins were expressed in Escherichia coli cells. The molecular weight of the beta type was approximately 100 kDa, and the alphas weighed approximately 60 kDa. Phylogenic tree construction revealed that the alphas could be classified into three known karyopherin-alpha subfamilies. We detected mRNA of the four karyopherins in normal 3rd day of 5th instar larvae, and in larvae injected with Gram-positive bacteria, Gram-negative bacteria, viruses, and fungi using real-time fluorescence quantitative reverse transcriptase-polymerase chain reaction, and found that the four karyopherins were widely distributed, but their expression levels were related to tissues type, the microbe injected, and the time point. PMID:26535618

  8. Silkworm Thorn Stem Extract Targets RSK2 and Suppresses Solar UV-Induced Cyclooxygenase-2 Expression.

    PubMed

    Kim, Jong-Eun; Lee, Ki Won

    2015-01-01

    Excessive exposure to solar UV (sUV) is associated with numerous human skin disorders, such as carcinogenesis, skin photoaging and skin inflammation. Silkworm Thorn (Cudraniatricuspidata, SW) is a plant belonging to the Moraceae family and widely present throughout Korea, China, and Japan. Most parts of the tree (including the fruit, leaf, stem, root, and bark) is consumable as a functional food or tea. In this study, we found that SW extract (SWE) inhibited the elevated expression of sUV-induced cyclooxygenase (COX)-2 levels in both HaCaT and JB6 cells. Levels of nuclear factor-κB and activator protein-1, two crucial transcription factors involved in COX-2 expression, were elevated by sUV treatment. Treatment with SWE abolished this activation. SWE also inhibited sUV-induced histone H3 phosphorylation. However, sUV-induced phosphorylation of Akt, c-Jun N-terminal kinase and p38 kinase remained unchanged in the presence of SWE. SWE inhibited RSK2 activity, and pull-down assays using SWE-Sepharose beads revealed that SWE binds directly with RSK2 in an ATP-competitive manner. These results suggest a potential for SWE to be developed as a cosmeceutical material and functional food constituent for the promotion of skin health.

  9. Molecular Expression of the Scribble Complex Genes, Dlg, Scrib and Lgl, in Silkworm, Bombyx mori.

    PubMed

    Qi, Hai-Sheng; Liu, Shu-Min; Li, Sheng; Wei, Zhao-Jun

    2013-05-30

    The Scribble protein complex genes, consisting of lethal giant larvae (Lgl), discs large (Dlg) and scribble (Scrib) genes, are components of an evolutionarily conserved genetic pathway that links the cell polarity in cells of humans and Drosophila. The tissue expression and developmental changes of the Scribble protein complex genes were documented using qRT-RCR method. The Lgl and Scrib genes could be detected in all the experimental tissues, including fat body, midgut, testis/ovary, wingdisc, trachea, malpighian tubule, hemolymph, prothoracic gland and silk gland. The Dlg gene, mainly expressed only in testis/ovary, could not be detected in prothoracic gland and hemolymph. In fat body, there were two higher expression stages of the three genes. The highest peak of the expression of the Lgl and Scrib genes in wingdisc lay at the 1st day of the 5th instar, but the Dlg gene was at 3rd day of 5th instar. The above results indicate that Scribble complex genes are involved in the process of molting and development of the wingdisc in the silkworm. This will be useful in the future for the elucidation of the detailed biological function of the three genes Scrib, Dlg and Lgl in B. mori.

  10. Knockout silkworms reveal a dispensable role for juvenile hormones in holometabolous life cycle.

    PubMed

    Daimon, Takaaki; Uchibori, Miwa; Nakao, Hajime; Sezutsu, Hideki; Shinoda, Tetsuro

    2015-08-01

    Insect juvenile hormones (JHs) prevent precocious metamorphosis and allow larvae to undergo multiple rounds of status quo molts. However, the roles of JHs during the embryonic and very early larval stages have not been fully understood. We generated and characterized knockout silkworms (Bombyx mori) with null mutations in JH biosynthesis or JH receptor genes using genome-editing tools. We found that embryonic growth and morphogenesis are largely independent of JHs in Bombyx and that, even in the absence of JHs or JH signaling, pupal characters are not formed in first- or second-instar larvae, and precocious metamorphosis is induced after the second instar at the earliest. We also show by mosaic analysis that a pupal specifier gene broad, which is dramatically up-regulated in the late stage of the last larval instar, is essential for pupal commitment in the epidermis. Importantly, the mRNA expression level of broad, which is thought to be repressed by JHs, remained at very low basal levels during the early larval instars of JH-deficient or JH signaling-deficient knockouts. Therefore, our study suggests that the long-accepted paradigm that JHs maintain the juvenile status throughout larval life should be revised because the larval status can be maintained by a JH-independent mechanism in very early larval instars. We propose that the lack of competence for metamorphosis during the early larval stages may result from the absence of an unidentified broad-inducing factor, i.e., a competence factor. PMID:26195792

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

    PubMed

    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.

  12. Equilibrium and kinetic adsorption study of a cationic dye by a natural adsorbent--silkworm pupa.

    PubMed

    Noroozi, B; Sorial, G A; Bahrami, H; Arami, M

    2007-01-01

    In this work the use of silkworm pupa, which is the waste of silk spinning industries has been investigated as an adsorbent for the removal of C.I. Basic Blue 41. The amino acid nature of the pupa provided a reasonable capability for dye removal. Equilibrium adsorption isotherms and kinetics were investigated. The adsorption equilibrium data were analyzed by using various adsorption isotherm models and the results have shown that adsorption behavior of the dye could be described reasonably well by either Langmuir or Freundlich models. The characteristic parameters for each isotherm have been determined. The monolayer adsorption capacity was determined to be 555 mg/g. Kinetic studies indicated that the adsorption follows pseudo-second-order kinetics with a rate constant of 0.0434 and 0.0572 g/min mg for initial dye concentration of 200 mg/l at 20 and 40 degrees C, respectively. Kinetic studies showed that film diffusion and intra-particle diffusion were simultaneously operating during the adsorption process. The rate constant for intra-particle diffusion was estimated to be 1.985 mg/g min(0.5).

  13. Durable grafting of silkworm pupa protein onto the surface of polyethylene terephthalate fibers.

    PubMed

    Zhou, Jianfeng; Zheng, Dandan; Zhang, Fengxiu; Zhang, Guangxian

    2016-12-01

    In this paper, reactive -NH2 groups (8.36×10(-6)mol/g fabric) were introduced to the surface of polyethylene terephthalate (PET) fabrics by a nitration and reduction method, and epoxy groups were introduced to silkworm pupa protein (SPP) by reaction with epoxy chloropropane. PET-SPP composite fabrics were then prepared by reaction of these two precursors. The results showed that the SPP was firmly grafted onto the PET fabric surface and that the hydrophilicity of the fabric was markedly improved by the grafting of SPP. SEM images revealed a layer of substance covering the surface of the PET fibers, and XPS investigation showed that the nitrogen content of the PET-SPP fabric was higher than that of the original PET fabric (2.32% vs 0%). ATR-FTIR adsorption bands at 1653 and 1543cm(-1) suggested the successful grafting of SPP onto the PET fabric surface. The DSC and TG of the PET fibers demonstrated that the thermal stability of the original PET fibers was maintained well by the SPP-grafted PET fibers. The breaking strength, bending rigidity, air permeability, and crease recovery angle of the original PET fabric were also retained by the SPP-grafted PET fabric.

  14. Effects of male silkworm pupa powder on the erectile dysfunction by chronic ethanol consumption in rats.

    PubMed

    Oh, Hong-Geun; Lee, Hak-Yong; Kim, Jung-Hoon; Kang, Young-Rye; Moon, Dea-In; Seo, Min-Young; Back, Hyang-Im; Kim, Sun-Young; Oh, Mi-Ra; Park, Soo-Hyun; Kim, Min-Gul; Jeon, Ji-Young; Shin, Sook-Jeong; Ryu, Kang-Sun; Chae, Soo-Wan; Kim, Okjin; Park, Jong-Kwan

    2012-06-01

    Erectile dysfunction (ED) is a highly prevalent disorder that affects millions of men worldwide. ED is now considered an early manifestation of atherosclerosis, and consequently, a precursor of systemic vascular disease. This study was designed to investigate the effects of male silkworm pupa powder (SWP) on the levels of nitric oxide synthase (NOS) expression, nitrite, and glutathione (GSH); lipid peroxidation; libido; and erectile response of the corpus cavernosum of the rat penis. We induced ED in the study animals by oral administration of 20% ethanol over 8 weeks. The SWP-treated male rats were divided into 3 groups that were orally administered 200, 400, and 800 mg/kg. The libido of the SWP-administered male rats was higher than that of the ethanol control group. In addition, the erectile response of the corpus cavernosum was restored in males on SWP administration, to a level similar to that of the normal group without ED. The testosterone concentration did not increase significantly. The lipid peroxidation in the corpus cavernosum of the male rats administered SWP decreased significantly. In contrast, compared to the ethanol group, SWP-administered male rats showed increased GSH levels in the corpus cavernosum. The level of nitrite and NOS expression in the corpus cavernosum of SWP-administered male rats increased significantly. These results indicated that SWP effectively restored ethanol-induced ED in male rats.

  15. Molecular characterization, localization, and distribution of innexins in the silkworm, Bombyx mori.

    PubMed

    Hong, Sun Mee; Noh, Si Kab; Kim, Kyung A; Mitsunobu, Hitoshi; Mon, Hiroaki; Lee, Jae Man; Kawaguchi, Yutaka; Kusakabe, Takahiro

    2009-09-01

    Gap junctions that allow for a direct exchange of second messenger and ions are the most conserved cellular structures in multicellular organisms. We have isolated and characterized a Bombyx mori gene innexin3 that encodes a new member of the innexin family required for the early embryonic development. The BmINX3 mRNA was 1,814 nucleotide residues in length, and the deduced amino acid sequence of BmInx3 shared 74% similarity with Apis melifera innexin3. The expression profile of the BmINX3 mRNA is similar to that of previously described BmINX2, expressed in ovary and testis after 5th instar larvae and in fat body after gut purge. However, during embryogenesis, the expression of BmINX3 mRNA is restricted to the blastokinesis stage. Microscopic observation of the BmInx2 and BmInx3 fused to fluorescent proteins showed an overlapping cytoplasmic expression, whereas the BmInx4 is accumulated in the cytoplasmic surface at which two cells have physical contact. This finding of innexins distribution in silkworm would provide an essential basis for future studies of the functions and interactions of innexins.

  16. Identification and Functional Analysis of the Pre-piRNA 3' Trimmer in Silkworms.

    PubMed

    Izumi, Natsuko; Shoji, Keisuke; Sakaguchi, Yuriko; Honda, Shozo; Kirino, Yohei; Suzuki, Tsutomu; Katsuma, Susumu; Tomari, Yukihide

    2016-02-25

    PIWI-interacting RNAs (piRNAs) play a crucial role in transposon silencing in animal germ cells. In piRNA biogenesis, single-stranded piRNA intermediates are loaded into PIWI-clade proteins and cleaved by Zucchini/MitoPLD, yielding precursor piRNAs (pre-piRNAs). Pre-piRNAs that are longer than the mature piRNA length are then trimmed at their 3' ends. Although recent studies implicated the Tudor domain protein Papi/Tdrkh in pre-piRNA trimming, the identity of Trimmer and its relationship with Papi/Tdrkh remain unknown. Here, we identified PNLDC1, an uncharacterized 3'-5' exonuclease, as Trimmer in silkworms. Trimmer is enriched in the mitochondrial fraction and binds to Papi/Tdrkh. Depletion of Trimmer and Papi/Tdrkh additively inhibits trimming, causing accumulation of ∼35-40-nt pre-piRNAs that are impaired for target cleavage and prone to degradation. Our results highlight the cooperative action of Trimmer and Papi/Tdrkh in piRNA maturation. PMID:26919431

  17. Mechanisms of nodule-specific melanization in the hemocoel of the silkworm, Bombyx mori.

    PubMed

    Shu, Min; Mang, Dingze; Fu, Gege Sun; Tanaka, Shiho; Endo, Haruka; Kikuta, Shingo; Sato, Ryoichi

    2016-03-01

    In the insect immune system, nodules are known to be a product of the cellular response against microorganisms and may be a preferential target for melanization. However, the mechanism of nodule-preferential melanization remains to be explored. In this study, we identified several mechanisms of nodule-preferential melanization by analyzing congregation and the activation of several factors involved in the prophenoloxidase (proPO)-activating system in the silkworm, Bombyx mori. Microorganism-binding assays revealed that B. mori larval plasma have an effective invading microorganism-surveillance network consisting of at least six pattern-recognition receptors (PRRs). We also found that a hemolymph serine proteinase, BmHP14, can bind to Saccharomyces cerevisiae. Pull-down assays showed that PRR C-type lectins form protein complexes with serine proteinase homologs, BmSPH1 and BmSPH2, which leads to the activated forms of BmSPH1 and BmSPH2 being gathered on microorganisms and trapped in nodules. Immunostaining analysis revealed that most factors in the proPO-activating system and some factors in the triggering system for antimicrobial peptide production exist in the granules of hemocytes which can gather in nodules. Western blot analysis showed that factors in the proPO-activating system are congregated in formed nodules by their concentration in plasma and aggregating hemocytes. PMID:26707571

  18. Redundant Mechanisms Recruit Actin into the Contractile Ring in Silkworm Spermatocytes

    PubMed Central

    Chen, Wei; Foss, Margit; Tseng, Kuo-Fu; Zhang, Dahong

    2008-01-01

    Cytokinesis is powered by the contraction of actomyosin filaments within the newly assembled contractile ring. Microtubules are a spindle component that is essential for the induction of cytokinesis. This induction could use central spindle and/or astral microtubules to stimulate cortical contraction around the spindle equator (equatorial stimulation). Alternatively, or in addition, induction could rely on astral microtubules to relax the polar cortex (polar relaxation). To investigate the relationship between microtubules, cortical stiffness, and contractile ring assembly, we used different configurations of microtubules to manipulate the distribution of actin in living silkworm spermatocytes. Mechanically repositioned, noninterdigitating microtubules can induce redistribution of actin at any region of the cortex by locally excluding cortical actin filaments. This cortical flow of actin promotes regional relaxation while increasing tension elsewhere (normally at the equatorial cortex). In contrast, repositioned interdigitating microtubule bundles use a novel mechanism to induce local stimulation of contractility anywhere within the cortex; at the antiparallel plus ends of central spindle microtubules, actin aggregates are rapidly assembled de novo and transported laterally to the equatorial cortex. Relaxation depends on microtubule dynamics but not on RhoA activity, whereas stimulation depends on RhoA activity but is largely independent of microtubule dynamics. We conclude that polar relaxation and equatorial stimulation mechanisms redundantly supply actin for contractile ring assembly, thus increasing the fidelity of cleavage. PMID:18767903

  19. Silkworm Thorn Stem Extract Targets RSK2 and Suppresses Solar UV-Induced Cyclooxygenase-2 Expression

    PubMed Central

    Kim, Jong-Eun; Lee, Ki Won

    2015-01-01

    Excessive exposure to solar UV (sUV) is associated with numerous human skin disorders, such as carcinogenesis, skin photoaging and skin inflammation. Silkworm Thorn (Cudraniatricuspidata, SW) is a plant belonging to the Moraceae family and widely present throughout Korea, China, and Japan. Most parts of the tree (including the fruit, leaf, stem, root, and bark) is consumable as a functional food or tea. In this study, we found that SW extract (SWE) inhibited the elevated expression of sUV-induced cyclooxygenase (COX)-2 levels in both HaCaT and JB6 cells. Levels of nuclear factor-κB and activator protein-1, two crucial transcription factors involved in COX-2 expression, were elevated by sUV treatment. Treatment with SWE abolished this activation. SWE also inhibited sUV-induced histone H3 phosphorylation. However, sUV-induced phosphorylation of Akt, c-Jun N-terminal kinase and p38 kinase remained unchanged in the presence of SWE. SWE inhibited RSK2 activity, and pull-down assays using SWE-Sepharose beads revealed that SWE binds directly with RSK2 in an ATP-competitive manner. These results suggest a potential for SWE to be developed as a cosmeceutical material and functional food constituent for the promotion of skin health. PMID:26506342

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

    PubMed

    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. PMID:27405010

  1. Advanced technologies for genetically manipulating the silkworm Bombyx mori, a model Lepidopteran insect.

    PubMed

    Xu, Hanfu; O'Brochta, David A

    2015-07-01

    Genetic technologies based on transposon-mediated transgenesis along with several recently developed genome-editing technologies have become the preferred methods of choice for genetically manipulating many organisms. The silkworm, Bombyx mori, is a Lepidopteran insect of great economic importance because of its use in silk production and because it is a valuable model insect that has greatly enhanced our understanding of the biology of insects, including many agricultural pests. In the past 10 years, great advances have been achieved in the development of genetic technologies in B. mori, including transposon-based technologies that rely on piggyBac-mediated transgenesis and genome-editing technologies that rely on protein- or RNA-guided modification of chromosomes. The successful development and application of these technologies has not only facilitated a better understanding of B. mori and its use as a silk production system, but also provided valuable experiences that have contributed to the development of similar technologies in non-model insects. This review summarizes the technologies currently available for use in B. mori, their application to the study of gene function and their use in genetically modifying B. mori for biotechnology applications. The challenges, solutions and future prospects associated with the development and application of genetic technologies in B. mori are also discussed.

  2. [Identification and characterization of a novel hemocyte specific gene Bm04862 in silkworm, Bombyx mori].

    PubMed

    Yu, Shuang; Su, Jingjing; Xu, Man; Zhang, Kui; Cui, Hongjuan

    2016-02-01

    Bm04862 is a novel hemocyte-specific gene, which has been identified and cloned through the microarray data in silkworm, Bombyx mori. Initially, we successfully obtained the full-length cDNA sequence of Bm04862 via the rapid amplification of cDNA ends (RACE). The sequence consists of an open reading frame (ORF) with 819 bp, which could encode 273 amino acid residues. The bioinformatics analysis predicted that Bm04862 was a transmembrane protein. Furthermore, qRT-PCR analysis revealed that the expression of Bm04862 was specifically detected in hemocyte and reached a peak at L4M and PP2 stage. Bm04862 was overexpressed in Sf9 insect cells, and the cellular localization indicated that Bm04862 was specifically located in cytoplasm and nuclei membrane. Interestingly, the expression of Bm04862 increas'd dramatically after challenged with Escherichia coli for 24 hours, which predicted its potential role in the innate immune system. Overall, this study could provide the fundamental knowledge for further research. PMID:27382774

  3. [Cloning and characterization of BmBrat in silkworm, Bombyx mori].

    PubMed

    Liang, Hanghua; Gao, Hongyan; Xu, Man; Tan, Peng; Cui, Hongjuan

    2016-03-01

    NHL proteins, which play important roles in regulation of cell proliferation and differentiation, have been extensively studied on mammals. Here, we cloned a member of NHL protein family namely BmBrat in silkworm. The full-length cDNA sequence of BmB rat was obtained by means of the rapid amplification of cDNA ends (RACE), including 3 614 bp. The ORF is 2 580 bp long, encoding a protein with 859 amino acid residues. The molecular weight is 94.3 kDa and the isoeledtric point (pI) is 6.65. The BmBrat expression profile was detected by RT-PCR at L5D3 larval stage, and it was expressed in all tissues, including silk gland, midgut, fat body and malpighian tubule. However, it was highly expressed in ovary and head. The expression profile was also detected at different stage of embryo development, and reached a peak at the 4th and 5th days of the embryonic period. Anti-BmBrat polyclonal antibody was generated f6llowing prokaryotic expression, protein purification and mice immunization, which is highly specific and effective for recognizing BmBrat protein through Western blotting and immunofluorescence staining. Subcellular localization of BmBrat in hemocytes revealed that it was specifically expressed in cytoplasm. This study provides a foundation for further research of the biological function of BmBrat gene. PMID:27349120

  4. Influence of RNAi knockdown for E-complex genes on the silkworm proleg development.

    PubMed

    Xiang, H; Li, M W; Guo, J H; Jiang, J H; Huang, Y P

    2011-01-01

    Larvae of many holometabolous insects possess abdominal appendages called prolegs. Lepidoptera larvae have prolegs in the segments A3-A6. Functions of Lepidoptera hox genes on these abdominal appendages development is still a controversial issue. In this article, we report the use of double strand RNA (dsRNA)-mediated interference (RNAi) to dissect the function of some hox genes, specifically E-complex genes Ubx, abd-A, and Abd-B, in the ventral appendage development of the Lepidoptera silkworm, Bombyx mori. We found that Ubx RNAi caused leg identity in A1 segment, abd-A RNAi caused severe defect of abdominal prolegs and Abd-B RNAi allowed proleg identity in more posterior abdominal segments. These results confirm that Lepidoptera hox genes Ubx and Abd-B have evolved the repressing function to ventral appendage development, which is similar to those of Drosophila. However, Lepidoptera abd-A might have been modified distinctively during evolution, and has important roles in directing the development of prolegs.

  5. Durable grafting of silkworm pupa protein onto the surface of polyethylene terephthalate fibers.

    PubMed

    Zhou, Jianfeng; Zheng, Dandan; Zhang, Fengxiu; Zhang, Guangxian

    2016-12-01

    In this paper, reactive -NH2 groups (8.36×10(-6)mol/g fabric) were introduced to the surface of polyethylene terephthalate (PET) fabrics by a nitration and reduction method, and epoxy groups were introduced to silkworm pupa protein (SPP) by reaction with epoxy chloropropane. PET-SPP composite fabrics were then prepared by reaction of these two precursors. The results showed that the SPP was firmly grafted onto the PET fabric surface and that the hydrophilicity of the fabric was markedly improved by the grafting of SPP. SEM images revealed a layer of substance covering the surface of the PET fibers, and XPS investigation showed that the nitrogen content of the PET-SPP fabric was higher than that of the original PET fabric (2.32% vs 0%). ATR-FTIR adsorption bands at 1653 and 1543cm(-1) suggested the successful grafting of SPP onto the PET fabric surface. The DSC and TG of the PET fibers demonstrated that the thermal stability of the original PET fibers was maintained well by the SPP-grafted PET fibers. The breaking strength, bending rigidity, air permeability, and crease recovery angle of the original PET fabric were also retained by the SPP-grafted PET fabric. PMID:27612829

  6. Lysine acetylation stabilizes SP2 protein in the silkworm Bombyx mori.

    PubMed

    Zhou, Yong; Wu, Chengcheng; Sheng, Qing; Jiang, Caiying; Chen, Qin; Lv, Zhengbing; Yao, Juming; Nie, Zuoming

    2016-01-01

    Lysine acetylation (Kac) is a vital post-translational modification that plays an important role in many cellular processes in organisms. In the present study, the nutrient storage proteins in hemolymph were first found to be highly acetylated-particularly SP2 protein, which contains 20 potential Kac sites. Further results confirmed that lysine acetylation could stabilize and up-regulate the protein level of anti-apoptosis protein SP2, thereby improving the survival of H2O2-treated BmN cells and suppressing the apoptosis induced by H2O2. The potential mechanism involved in the inhibition of ubiquitin-mediated proteasomal degradation by crosstalk between lysine acetylation and ubiquitination. Our results showed that the increase in the acetylation level by TSA could decrease the ubiquitination and improve the protein level of SP2, indicating that lysine acetylation could influence the SP2 protein level through competition between ubiquitination and the suppression of ubiquitin-mediated proteasomal degradation, thereby stabilizing the protein. SP2 is a major nutrient storage protein from hemolymph for amino acid storage and utilization. The crosstalk between lysine acetylation and ubiquitination of SP2 might imply an important role of lysine acetylation for nutrient storage and utilization in silkworm. PMID:27374983

  7. Midgut epithelium in molting silkworm: A fine balance among cell growth, differentiation, and survival.

    PubMed

    Franzetti, Eleonora; Casartelli, Morena; D'Antona, Paola; Montali, Aurora; Romanelli, Davide; Cappellozza, Silvia; Caccia, Silvia; Grimaldi, Annalisa; de Eguileor, Magda; Tettamanti, Gianluca

    2016-07-01

    The midgut of insects has attracted great attention as a system for studying intestinal stem cells (ISCs) as well as cell death-related processes, such as apoptosis and autophagy. Among insects, Lepidoptera represent a good model to analyze these cells and processes. In particular, larva-larva molting is an interesting developmental phase since the larva must deal with nutrient starvation and its organs are subjected to rearrangements due to proliferation and differentiation events. Several studies have analyzed ISCs in vitro and characterized key factors involved in their division and differentiation during molt. However, in vivo studies performed during larva-larva transition on these cells, and on the whole midgut epithelium, are fragmentary. In the present study, we analyzed the larval midgut epithelium of the silkworm, Bombyx mori, during larva-larva molting, focusing our attention on ISCs. Moreover, we investigated the metabolic changes that occur in the epithelium and evaluated the intervention of autophagy. Our data on ISCs proliferation and differentiation, autophagy activation, and metabolic and functional activities of the midgut cells shed light on the complexity of this organ during the molting phase. PMID:27349418

  8. Proteins in the Cocoon of Silkworm Inhibit the Growth of Beauveria bassiana.

    PubMed

    Guo, Xiaomeng; Dong, Zhaoming; Zhang, Yan; Li, Youshan; Liu, Huawei; Xia, Qingyou; Zhao, Ping

    2016-01-01

    Silk cocoons are composed of fiber proteins (fibroins) and adhesive glue proteins (sericins), which provide a physical barrier to protect the inside pupa. Moreover, other proteins were identified in the cocoon silk, many of which are immune related proteins. In this study, we extracted proteins from the silkworm cocoon by Tris-HCl buffer (pH7.5), and found that they had a strong inhibitory activity against fungal proteases and they had higher abundance in the outer cocoon layers than in the inner cocoon layers. Moreover, we found that extracted cocoon proteins can inhibit the germination of Beauveria bassiana spores. Consistent with the distribution of protease inhibitors, we found that proteins from the outer cocoon layers showed better inhibitory effects against B. bassiana spores than proteins from the inner layers. Liquid chromatography-tandem mass spectrometry was used to reveal the extracted components in the scaffold silk, the outermost cocoon layer. A total of 129 proteins were identified, 30 of which were annotated as protease inhibitors. Protease inhibitors accounted for 89.1% in abundance among extracted proteins. These protease inhibitors have many intramolecular disulfide bonds to maintain their stable structure, and remained active after being boiled. This study added a new understanding to the antimicrobial function of the cocoon. PMID:27032085

  9. Hydrologic data summary for the White Oak Watershed at Oak Ridge National Laboratory, Oak Ridge, Tennessee, October 1990--December 1991

    SciTech Connect

    Borders, D.M.; Gregory, S.M.; Clapp, R.B.; Frederick, B.J.; Watts, J.A.

    1992-06-01

    This report summarizes for the 15-month period of October 1990-- December 1991 the available dynamic hydrologic data collected, primarily on the White Oak Creek (WOC) watershed, along with information collected on the surface flow systems that affect the quality or quantity of surface water. The collection of hydrologic data is one component of numerous, ongoing Oak Ridge National Laboratory (ORNL) environmental studies and monitoring programs and is intended to: (1) characterize the quantity and quality of water in the flow systems; (2) assist with the planning and assessment of remedial action activities; and, (3) provide long-term availability of data and quality assurance. Characterization of the hydrology of the WOC watershed is critical for understanding the processes that drive contaminant transport in the watershed. Identification of spatial and temporal trends in hydrologic parameters and mechanisms that affect the movement of contaminants supports the development of interim corrective measures and remedial restoration alternatives. In addition, hydrologic monitoring supports long-term assessment of the effectiveness of remedial actions in limiting the transport of contaminants across Waste Area Grouping (WAG) boundaries and ultimately to the off-site environment. For these reasons, it is of paramount importance to the Environmental Restoration Program (ERP) to collect and report hydrologic data activities that contribute to the Site Investigations component of the ERP. (White Oak Creek is also referred to as ``Whiteoak`` Creek).

  10. Hydrologic data summary for the White Oak Watershed at Oak Ridge National Laboratory, Oak Ridge, Tennessee, October 1990--December 1991

    SciTech Connect

    Borders, D.M.; Gregory, S.M.; Clapp, R.B.; Frederick, B.J.; Watts, J.A.

    1992-06-01

    This report summarizes for the 15-month period of October 1990-- December 1991 the available dynamic hydrologic data collected, primarily on the White Oak Creek (WOC) watershed, along with information collected on the surface flow systems that affect the quality or quantity of surface water. The collection of hydrologic data is one component of numerous, ongoing Oak Ridge National Laboratory (ORNL) environmental studies and monitoring programs and is intended to: (1) characterize the quantity and quality of water in the flow systems; (2) assist with the planning and assessment of remedial action activities; and, (3) provide long-term availability of data and quality assurance. Characterization of the hydrology of the WOC watershed is critical for understanding the processes that drive contaminant transport in the watershed. Identification of spatial and temporal trends in hydrologic parameters and mechanisms that affect the movement of contaminants supports the development of interim corrective measures and remedial restoration alternatives. In addition, hydrologic monitoring supports long-term assessment of the effectiveness of remedial actions in limiting the transport of contaminants across Waste Area Grouping (WAG) boundaries and ultimately to the off-site environment. For these reasons, it is of paramount importance to the Environmental Restoration Program (ERP) to collect and report hydrologic data activities that contribute to the Site Investigations component of the ERP. (White Oak Creek is also referred to as Whiteoak'' Creek).

  11. Real-Time Mass Spectrometry Monitoring of Oak Wood Toasting: Elucidating Aroma Development Relevant to Oak-aged Wine Quality.

    PubMed

    Farrell, Ross R; Wellinger, Marco; Gloess, Alexia N; Nichols, David S; Breadmore, Michael C; Shellie, Robert A; Yeretzian, Chahan

    2015-11-27

    We introduce a real-time method to monitor the evolution of oak aromas during the oak toasting process. French and American oak wood boards were toasted in an oven at three different temperatures, while the process-gas was continuously transferred to the inlet of a proton-transfer-reaction time-of-flight mass spectrometer for online monitoring. Oak wood aroma compounds important for their sensory contribution to oak-aged wine were tentatively identified based on soft ionization and molecular mass. The time-intensity profiles revealed toasting process dynamics illustrating in real-time how different compounds evolve from the oak wood during toasting. Sufficient sensitivity was achieved to observe spikes in volatile concentrations related to cracking phenomena on the oak wood surface. The polysaccharide-derived compounds exhibited similar profiles; whilst for lignin-derived compounds eugenol formation differed from that of vanillin and guaiacol at lower toasting temperatures. Significant generation of oak lactone from precursors was evident at 225 (o)C. Statistical processing of the real-time aroma data showed similarities and differences between individual oak boards and oak wood sourced from the different origins. This study enriches our understanding of the oak toasting process and demonstrates a new analytical approach for research on wood volatiles.

  12. Real-Time Mass Spectrometry Monitoring of Oak Wood Toasting: Elucidating Aroma Development Relevant to Oak-aged Wine Quality

    PubMed Central

    Farrell, Ross R.; Wellinger, Marco; Gloess, Alexia N.; Nichols, David S.; Breadmore, Michael C.; Shellie, Robert A.; Yeretzian, Chahan

    2015-01-01

    We introduce a real-time method to monitor the evolution of oak aromas during the oak toasting process. French and American oak wood boards were toasted in an oven at three different temperatures, while the process-gas was continuously transferred to the inlet of a proton-transfer-reaction time-of-flight mass spectrometer for online monitoring. Oak wood aroma compounds important for their sensory contribution to oak-aged wine were tentatively identified based on soft ionization and molecular mass. The time-intensity profiles revealed toasting process dynamics illustrating in real-time how different compounds evolve from the oak wood during toasting. Sufficient sensitivity was achieved to observe spikes in volatile concentrations related to cracking phenomena on the oak wood surface. The polysaccharide-derived compounds exhibited similar profiles; whilst for lignin-derived compounds eugenol formation differed from that of vanillin and guaiacol at lower toasting temperatures. Significant generation of oak lactone from precursors was evident at 225 oC. Statistical processing of the real-time aroma data showed similarities and differences between individual oak boards and oak wood sourced from the different origins. This study enriches our understanding of the oak toasting process and demonstrates a new analytical approach for research on wood volatiles. PMID:26610612

  13. Real-Time Mass Spectrometry Monitoring of Oak Wood Toasting: Elucidating Aroma Development Relevant to Oak-aged Wine Quality

    NASA Astrophysics Data System (ADS)

    Farrell, Ross R.; Wellinger, Marco; Gloess, Alexia N.; Nichols, David S.; Breadmore, Michael C.; Shellie, Robert A.; Yeretzian, Chahan

    2015-11-01

    We introduce a real-time method to monitor the evolution of oak aromas during the oak toasting process. French and American oak wood boards were toasted in an oven at three different temperatures, while the process-gas was continuously transferred to the inlet of a proton-transfer-reaction time-of-flight mass spectrometer for online monitoring. Oak wood aroma compounds important for their sensory contribution to oak-aged wine were tentatively identified based on soft ionization and molecular mass. The time-intensity profiles revealed toasting process dynamics illustrating in real-time how different compounds evolve from the oak wood during toasting. Sufficient sensitivity was achieved to observe spikes in volatile concentrations related to cracking phenomena on the oak wood surface. The polysaccharide-derived compounds exhibited similar profiles; whilst for lignin-derived compounds eugenol formation differed from that of vanillin and guaiacol at lower toasting temperatures. Significant generation of oak lactone from precursors was evident at 225 oC. Statistical processing of the real-time aroma data showed similarities and differences between individual oak boards and oak wood sourced from the different origins. This study enriches our understanding of the oak toasting process and demonstrates a new analytical approach for research on wood volatiles.

  14. Impacts on silkworm larvae midgut proteomics by transgenic Trichoderma strain and analysis of glutathione S-transferase sigma 2 gene essential for anti-stress response of silkworm larvae.

    PubMed

    Li, Yingying; Dou, Kai; Gao, Shigang; Sun, Jianan; Wang, Meng; Fu, Kehe; Yu, Chuanjin; Wu, Qiong; Li, Yaqian; Chen, Jie

    2015-08-01

    Lepidoptera is a large order of insects that have major impacts on humans as agriculture pests. The midgut is considered an important target for insect control. In the present study, 10 up-regulated, 18 down-regulated, and one newly emerged protein were identified in the transgenic Trichoderma-treated midgut proteome. Proteins related to stress response, biosynthetic process, and metabolism process were further characterized through quantitative real-time PCR (qPCR). Of all the identified proteins, the glutathione S-transferase sigma 2 (GSTs2) gene displayed enhanced expression when larvae were fed with Trichoderma wild-type or transgenic strains. Down regulation of GSTs2 expression by RNA interference (RNAi) resulted in inhibition of silkworm growth when larvae were fed with mulberry leaves treated with the transgenic Trichoderma strain. Weight per larva decreased by 18.2%, 11.9%, and 10.7% in the untreated control, ddH2O, and GFP dsRNA groups, respectively, at 24h, while the weight decrease was higher at 42.4%, 28.8% and 32.4% at 72 h after treatment. Expression of glutathione S-transferase omega 2 (GSTo2) was also enhanced when larvae were fed with mulberry leaves treated with the transgenic Trichoderma strain. These results indicated that there was indeed correlation between enhanced expression of GSTs2 and the anti-stress response of silkworm larvae against Trichoderma. This study represents the first attempt at understanding the effects of transgenic organisms on the midgut proteomic changes in silkworm larvae. Our findings could not only broaden the biological control targets of insect at the molecular level, but also provide a theoretical foundation for biological safety evaluation of the transgenic Trichoderma strain.

  15. A Review of Polyphenolics in Oak Woods

    PubMed Central

    Zhang, Bo; Cai, Jian; Duan, Chang-Qing; Reeves, Malcolm J.; He, Fei

    2015-01-01

    Polyphenolics, which are ubiquitous in plants, currently are among the most studied phytochemicals because of their perceptible chemical properties and antioxidant activity. Oak barrels and their alternatives, which are widely used in winemaking nowadays, contribute polyphenolics to wines and are thought to play crucial roles in the development of wines during aging. This study summarizes the detailed information of polyphenolics in oak woods and their products by examining their structures and discussing their chemical reactions during wine aging. This paper evaluates the most recent developments in polyphenolic chemistry by summarizing their extraction, separation, and their identification by the use of chromatographic and spectral techniques. In addition, this paper also introduces polyphenol bioactive ingredients in other plant foods. PMID:25826529

  16. VOC emissions from beech, birch, and oak

    NASA Astrophysics Data System (ADS)

    Wildt, J.; Folkers, A.; Koch, N.; Kleist, E.

    2003-04-01

    VOC emissions from beech (Fagus sylvatica), birch (Betula pendula), and oak (Quercus robur) were studied in continuously stirred tank reactors. Oak emitted nearly exclusively isoprene. The dependence of these isoprene emissions on temperature and photosynthetic radiation (PAR) could quite well be described with existing algorithms and the emission factors were fairly constant. Beech and birch emitted mainly short chained oxygenated VOC and monoterpenes. Temperature and PAR dependence of monoterpene emissions were superimposed by a slow frequency modulation. Hence, descriptions of these emissions with existing algorithms were not successful. Moreover, in some cases the emission pattern switched drastically. For birch it was observed that the plant switched from a sesquiterpene emitter to a monoterpene emitter. emission pattern plants. Emissions of ethanol, acetaldehyde, and methanol were not affected by PAR. Here, the emission factors are determined by other factors not included in existing algorithms.

  17. Oak Ridge National Laboratory Waste Management Plan

    SciTech Connect

    Not Available

    1992-12-01

    The objective of the Oak Ridge National Laboratory Waste Management Plan is to compile and to consolidate information annually on how the ORNL Waste Management Program is conducted, which waste management facilities are being used to manage wastes, what forces are acting to change current waste management systems, what activities are planned for the forthcoming fiscal year (FY), and how all of the activities are documented.

  18. ORNL (Oak Ridge National Laboratory) 89

    SciTech Connect

    Anderson, T.D.; Appleton, B.R.; Jefferson, J.W.; Merriman, J.R.; Mynatt, F.R.; Richmond, C.R.; Rosenthal, M.W.

    1989-01-01

    This is the inaugural issues of an annual publication about the Oak Ridge National Laboratory. Here you will find a brief overview of ORNL, a sampling of our recent research achievements, and a glimpse of the directions we want to take over the next 15 years. A major purpose of ornl 89 is to provide the staff with a sketch of the character and dynamics of the Laboratory.

  19. AmeriFlux US-Oho Oak Openings

    SciTech Connect

    Chen, Jiquan

    2016-01-01

    This is the AmeriFlux version of the carbon flux data for the site US-Oho Oak Openings. Site Description - The Ohio Oak Openings site is located within the Oak Openings Preserve Metropark of northwest Ohio, one of the few remaining oak woodlands/savanna/prairie complexes in the Midwest. Declared one of the "One of America's Last Great Places" by the Nature Conservancy the area consists of four main vegetation types: Oak Woodlands, Oak Savanna, Floodplain Forests and Wet Prairies. The stand surrounding the tower is mainly Oak Woodlands dominated by red, white and black oaks with a relatively abundant population of red maples indicating high soil moisture retention and a history of limited fire disturbances. Most of the area was cleared for agriculture at the time of Euro-American settlements in the mid to late-19th century. A large fraction of the cleared land was later abandoned due to the poor sandy soils. These areas reverted to Oak Savannas and in cases where fire was limited progressively made the transition to Oak Woodlands. Today patches of the forest are burned every few years as part of prescribed burning cycle to control stand density.

  20. A 1H-NMR Based Study on Hemolymph Metabolomics in Eri Silkworm after Oral Administration of 1-Deoxynojirimycin

    PubMed Central

    Deng, Ming-Jie; Lin, Xiao-Dong; Lin, Qiu-Ting; Wen, De-Fu; Zhang, Mei-Ling; Wang, Xian-Qin; Gao, Hong-Chang; Xu, Jia-Ping

    2015-01-01

    We aimed to investigate whether 1-deoxynojirimycin (DNJ) modulates glycometabolism and has toxicity in Eri silkworm (Samia cynthia ricini, Saturniidae). In this paper, hemolymph metabolites were used to explore metabolic changes after oral administration of DNJ or mulberry latex and to characterize the biological function of DNJ at the metabolic and systemic levels. Hemolymph samples were collected from fourth-instar larvae of Eri silkworm and ex-vivo high-resolution 1H nuclear magnetic resonance (NMR) spectra were acquired from the collected hemolymph samples. Then the obtained spectra were analyzed by principal component analysis (PCA) and independent-samples t-test. Metabolic pattern recognition analysis of hemolymph samples indicated that the groups of 0.25% DNJ, latex, and the mixture of 0.5% DNJ and latex (1:1) were significantly different from the control group. Moreover, compared to the control group, the groups of 0.25% DNJ, latex, and the mixture of 0.5% DNJ and latex (1:1) showed the decreased levels of citrate, succinate, fumarate, malate, and glutamine in hemolymph, the groups of 0.25% DNJ and the mixture of 0.5% DNJ and latex (1:1) showed the increased levels of trehalose and lactate. In addition, mulberry leaves exude latex was highly toxic to Eri silkworm because rich unidentified high-molecular-weight factor (s) acted as toxic substances. In our results, latex caused 20 deaths among 50 fourth-instar larvae of Eri silkmoth, but DNJ or the mixture did not caused death. All these results suggest that DNJ has a positive impact on the reverse glycometabolism by modulating glycometabolism and inhibiting glucogenesis and energy metabolism. DNJ is a secure substance as a single-ingredient antidiabetic medicine due to its nontoxicity and bioactivity. PMID:26148185

  1. Recent progress in development of transgenic silkworms overexpressing recombinant human proteins with therapeutic potential in silk glands.

    PubMed

    Itoh, Kohji; Kobayashi, Isao; Nishioka, So-Ichiro; Sezutsu, Hideki; Machii, Hiroaki; Tamura, Toshiki

    2016-02-01

    Since 2000, transgenic silkworms have been developed to produce recombinant proteins with therapeutic potential for future clinical use, including antibody preparations. Lysosomal storage diseases (LSDs) are inherited metabolic disorders caused by mutations of lysosomal enzymes associated with excessive accumulation of natural substrates and neurovisceral symptoms. Over the past few years, enzyme replacement therapy (ERT) with human lysosomal enzymes produced by genetically engineered mammalian cell lines has been used clinically to treat several patients with an LSD involving multi-organ symptoms. ERT is based on the incorporation of recombinant glycoenzymes by their binding to glycan receptors on the surface of target cells and their subsequent delivery to lysosomes. However, ERT has several disadvantages, including difficulty mass producing human enzymes, dangers of pathogen contamination, and high costs. Recently, the current authors have succeeded in producing transgenic silkworms overexpressing human lysosomal enzymes in the silk glands and the authors have purified catalytically active enzymes from the middle silk glands. Silk gland-derived human enzymes carrying high-mannose and pauci-mannose N-glycans were endocytosed by monocytes via the mannose receptor pathway and were then delivered to lysosomes. Conjugates with cell-penetrating peptides were also taken up by cultured fibroblasts derived from patients with enzyme deficiencies to restore intracellular catalytic activity and reduce the excessive accumulation of substrates in patient fibroblasts. Transgenic silkworms overexpressing human lysosomal enzymes in the silk glands could serve as future bioresources that provide safe therapeutic enzymes for the treatment of LSDs. Combining recent developments in transglycosylation technology with microbial endoglycosidases will promote the development of therapeutic glycoproteins as bio-medicines. PMID:26971553

  2. Molecular Signatures of Reduced Nerve Toxicity by CeCl3 in Phoxim-exposed Silkworm Brains

    PubMed Central

    Wang, Binbin; Li, Fanchi; Ni, Min; Zhang, Hua; Xu, Kaizun; Tian, Jianghai; Hu, Jingsheng; Shen, Weide; Li, Bing

    2015-01-01

    CeCl3 can reduce the damage caused by OP pesticides, in this study we used the brain of silkworms to investigate the mechanism of CeCl3 effects on pesticide resistance. The results showed that phoxim treatments led to brain damages, swelling and death of neurons, chromatin condensation, and mitochondrial damage. Normal nerve conduction was severely affected by phoxim treatments, as revealed by: increases in the contents of neurotransmitters Glu, NO, and ACh by 63.65%, 61.14%, and 98.54%, respectively; decreases in the contents of 5-HT and DA by 53.19% and 43.71%, respectively; reductions in the activities of Na+/K+-ATPase, Ca2+/Mg2+-ATPase, and AChE by 85.27%, 85.63%, and 85.63%, respectively; and increase in the activity of TNOS by 22.33%. CeCl3 pretreatment can significantly reduce such damages. Results of DGE and qRT-PCR indicated that CeCl3 treatments significantly upregulated the expression levels of CYP4G23, cyt-b5, GSTs-σ1, ace1, esterase-FE4, and β-esterase 2. Overall, phoxim treatments cause nerve tissue lesions, neuron death, and nerve conduction hindrance, but CeCl3 pretreatments can promote the expression of phoxim resistance-related genes in silkworm brains to reduce phoxim-induced damages. Our study provides a potential new method to improve the resistance of silkworms against OP pesticides. PMID:26227613

  3. Floodplain and wetlands assessment of the White Oak Creek Embayment

    SciTech Connect

    Not Available

    1991-07-01

    This report describes the proposed methods for dealing with contaminants that have accumulated in White Oak Creek, White Oak Lake, and the White Oak Creek Embayment as a result of process releases and discharges from the Oak Ridge National Laboratory. Alternative methods of cleaning up the area which were considered in accordance with regulatory guidelines are listed, and information supporting the selected methods is provided. Also included are results of a site survey conducted at the White Oak Creek Embayment and the expected effects of the proposed control structures on the floodplain and wetlands. The appendix contains figures showing the nine cross-sections of the stream channel surveyed during studies of the White Oak Creek area.

  4. Oak moss extracts in the diagnosis of fragrance contact allergy.

    PubMed

    Johansen, Jeanne Duus; Heydorn, Siri; Menné, Torkil

    2002-03-01

    Oak moss absolute is one of the eight ingredients of the fragrance mix (FM) used for diagnosing perfume allergy. Oak moss absolute is an extract prepared from the lichen Evernia prunastri growing on oak trees. It has been shown that the oak moss patch test material from one producer contained resin acids which are ingredients of another lichen, tree moss. Resin acids, e.g. abietic acid and dehydroabietic acid, are also the main allergens in colophonium. The aim of the study was to assess whether the contamination of oak moss absolute and thus the FM with resin acids had affected their diagnostic value so that they, instead of indicating fragrance allergy, had become indicators of allergy to resin acids and thus colophonium. Two studies were undertaken. First the relationship between patch test reactions to FM, oak moss absolute, both with contents of resin acids, and colophonium were assessed in 885 consecutive patients. A significant relationship between reactions to colophonium and FM was seen (p < 0.001) as well as a significant relationship between oak moss absolute and colophonium (p < 0.001). The relationship between colophonium and FM was still significant when all reactions to oak moss absolute were disregarded (p < 0.001), showing a relationship also between colophonium and fragrance ingredients other than oak moss absolute. Second, 119 consecutive patients were tested with an old and a new version of oak moss absolute containing resin acid (0.05%) and no measurable resin acid, respectively, and with the corresponding FM. No overall difference in reactivity to the old and new version of oak moss absolute/FM was seen. It is concluded the diagnostic value of oak moss absolute as indicator fragrance contact allergy has been and is unaffected by the resin acid contamination.

  5. Identification of two juvenile hormone inducible transcription factors from the silkworm, Bombyx mori.

    PubMed

    Matsumoto, Hitoshi; Ueno, Chihiro; Nakamura, Yuki; Kinjoh, Terunori; Ito, Yuka; Shimura, Sachiko; Noda, Hiroaki; Imanishi, Shigeo; Mita, Kazuei; Fujiwara, Haruhiko; Hiruma, Kiyoshi; Shinoda, Tetsuro; Kamimura, Manabu

    2015-09-01

    Juvenile hormone (JH) regulates many physiological processes in insects. However, the signal cascades in which JH is active have not yet been fully elucidated, particularly in comparison to another major hormone ecdysteroid. Here we identified two JH inducible transcription factors as candidate components of JH signaling pathways in the silkworm, Bombyx mori. DNA microarray analysis showed that expression of two transcription factor genes, E75 and Enhancer of split mβ (E(spl)mβ), was induced by juvenile hormone I (JH I) in NIAS-Bm-aff3 cells. Real time RT-PCR analysis confirmed that expression of four E75 isoforms (E75A, E75B, E75C and E75D) and E(spl)mβ was 3-8 times greater after JH I addition. Addition of the protein synthesis inhibitor cycloheximide did not suppress JH-induced expression of the genes, indicating that they were directly induced by JH. JH-induced expression of E75 and E(spl)mβ was also observed in four other B. mori cell lines and in larval hemocytes of final instar larvae. Notably, E75A expression was induced very strongly in larval hemocytes by topical application of the JH analog fenoxycarb; the level of induced expression was comparable to that produced by feeding larvae with 20-hydroxyecdysone. These results suggest that E75 and E(spl)mβ are general and direct target genes of JH and that the transcription factors encoded by these genes play important roles in JH signaling.

  6. Identification of C-type lectin-domain proteins (CTLDPs) in silkworm Bombyx mori.

    PubMed

    Rao, Xiang-Jun; Shahzad, Toufeeq; Liu, Su; Wu, Peng; He, Yan-Ting; Sun, Wei-Jia; Fan, Xiang-Yun; Yang, Yun-Fan; Shi, Qiao; Yu, Xiao-Qiang

    2015-12-01

    C-type lectins (CTLs) represent a large family of proteins that can bind carbohydrate moieties normally in a calcium-dependent manner. CTLs play important roles in mediating cell adhesion and the recognition of pathogens in the immune system. In the present study, we have identified 23 CTL genes in domestic silkworm Bombyx mori. CTL-domain proteins (CTLDPs) are classified into three groups based on the number of carbohydrate-recognition domains (CRDs) and the domain architectures. These include twelve CTL-S (Single-CRD), six immulectins (Dual-CRD) and five CTL-X (CRD with other domains). We studied their phylogenetic features, analyzed the conserved residues, predicted tertiary structures, and examined the tissue expression profile and immune inducibility. Through bioinformatics analysis, we have putatively identified ten secretory and two cytoplasmic CTL-S; four secretory and two cytoplasmic immulectins; one secretory, one cytoplasmic and three transmembrane forms of CTL-X. Most B. mori CTLDPs form monophyletic groups with orthologs from Lepidoptera, Diptera, Coleoptera and Hymenoptera species. Immulectins of B. mori and Manduca sexta evolved from common ancestor genes perhaps due to gene duplication events of CTL-S ancestor genes. Homology modeling revealed that the overall structures of B. mori CTL domains are analogous to those of humans with a variable loop region. We examined the expression profile of CTLDP genes in naïve and immune-stimulated tissues. The expression and induction of CTLDP genes were related to the tissues and microorganisms. Together, our gene identification, sequence comparison, phylogenetic analysis, homology modeling and expression analysis laid a good foundation for the further studies of B. mori CTLDPs and comparative genomics.

  7. Identification of C-type lectin-domain proteins (CTLDPs) in silkworm Bombyx mori.

    PubMed

    Rao, Xiang-Jun; Shahzad, Toufeeq; Liu, Su; Wu, Peng; He, Yan-Ting; Sun, Wei-Jia; Fan, Xiang-Yun; Yang, Yun-Fan; Shi, Qiao; Yu, Xiao-Qiang

    2015-12-01

    C-type lectins (CTLs) represent a large family of proteins that can bind carbohydrate moieties normally in a calcium-dependent manner. CTLs play important roles in mediating cell adhesion and the recognition of pathogens in the immune system. In the present study, we have identified 23 CTL genes in domestic silkworm Bombyx mori. CTL-domain proteins (CTLDPs) are classified into three groups based on the number of carbohydrate-recognition domains (CRDs) and the domain architectures. These include twelve CTL-S (Single-CRD), six immulectins (Dual-CRD) and five CTL-X (CRD with other domains). We studied their phylogenetic features, analyzed the conserved residues, predicted tertiary structures, and examined the tissue expression profile and immune inducibility. Through bioinformatics analysis, we have putatively identified ten secretory and two cytoplasmic CTL-S; four secretory and two cytoplasmic immulectins; one secretory, one cytoplasmic and three transmembrane forms of CTL-X. Most B. mori CTLDPs form monophyletic groups with orthologs from Lepidoptera, Diptera, Coleoptera and Hymenoptera species. Immulectins of B. mori and Manduca sexta evolved from common ancestor genes perhaps due to gene duplication events of CTL-S ancestor genes. Homology modeling revealed that the overall structures of B. mori CTL domains are analogous to those of humans with a variable loop region. We examined the expression profile of CTLDP genes in naïve and immune-stimulated tissues. The expression and induction of CTLDP genes were related to the tissues and microorganisms. Together, our gene identification, sequence comparison, phylogenetic analysis, homology modeling and expression analysis laid a good foundation for the further studies of B. mori CTLDPs and comparative genomics. PMID:26187302

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

    PubMed

    Daimon, Takaaki; Kozaki, Toshinori; Niwa, Ryusuke; Kobayashi, Isao; Furuta, Kenjiro; Namiki, Toshiki; Uchino, Keiro; Banno, Yutaka; Katsuma, Susumu; Tamura, Toshiki; Mita, Kazuei; Sezutsu, Hideki; Nakayama, Masayoshi; Itoyama, Kyo; Shimada, Toru; Shinoda, Tetsuro

    2012-01-01

    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.

  9. Precocious Metamorphosis in the Juvenile Hormone–Deficient Mutant of the Silkworm, Bombyx mori

    PubMed Central

    Daimon, Takaaki; Kozaki, Toshinori; Niwa, Ryusuke; Kobayashi, Isao; Furuta, Kenjiro; Namiki, Toshiki; Uchino, Keiro; Banno, Yutaka; Katsuma, Susumu; Tamura, Toshiki; Mita, Kazuei; Sezutsu, Hideki; Nakayama, Masayoshi; Itoyama, Kyo; Shimada, Toru; Shinoda, Tetsuro

    2012-01-01

    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. PMID:22412378

  10. Shotgun proteomic analysis of the fat body during metamorphosis of domesticated silkworm (Bombyx mori).

    PubMed

    Yang, Huijuan; Zhou, Zhonghua; Zhang, Huarong; Chen, Ming; Li, Jianying; Ma, Yingying; Zhong, Boxiong

    2010-05-01

    Protein expression profiles in the fat bodies of larval, pupal, and moth stages of silkworm were determined using shotgun proteomics and MS sequencing. We identified 138, 217, and 86 proteins from the larval, pupal and moth stages, respectively, of which 12 were shared by the 3 stages. There were 92, 150, and 45 specific proteins identified in the larval, pupal and moth stages, respectively, of which 17, 68, and 9 had functional annotations. Among the specific proteins identified in moth fat body, sex-specific storage-protein 1 precursor and chorion protein B8 were unique to the moth stage, indicating that the moth stage fat body is more important for adult sexual characteristics. Many ribosomal proteins (L23, L4, L5, P2, S10, S11, S15A and S3) were found in pupal fat bodies, whereas only three (L14, S20, and S7) and none were identified in larval and moth fat bodies, respectively. Twenty-three metabolic enzymes were identified in the pupal stage, while only four and two were identified in the larval and moth stages, respectively. In addition, an important protein, gloverin2, was only identified in larval fat bodies. Gene ontology (GO) analysis of the proteins specific to the three stages linked them to the cellular component, molecular function, and biological process categories. The most diverse GO functional classes were involved by the relatively less specific proteins identified in larva. GO analysis of the proteins shared among the three stages showed that the pupa and moth stages shared the most similar protein functions in the fat body.

  11. Effect of TiO2 nanoparticles on the reproduction of silkworm.

    PubMed

    Ni, Min; Li, Fanchi; Wang, Binbin; Xu, Kaizun; Zhang, Hua; Hu, Jingsheng; Tian, Jianghai; Shen, Weide; Li, Bing

    2015-03-01

    Silkworm (Bombyx mori) is an important economic insect and the model insect of Lepidoptera. Because of its high fecundity and short reproduction cycle, it has been widely used in reproduction and development research. The high concentrations of titanium dioxide nanoparticles (TiO2 NPs) show reproductive toxicity, while low concentrations of TiO2 NPs have been used as feed additive and demonstrated significant biological activities. However, whether the low concentrations of TiO2 NPs affect the reproduction of B. mori has not been reported. In this study, the growth and development of gonad of B. mori fed with a low concentration of TiO2 NPs (5 mg/L) were investigated by assessing egg production and expression of reproduction-related genes. The results showed that the low concentration of TiO2 NPs resulted in faster development of the ovaries and testes and more gamete differentiation and formation, with an average increase of 51 eggs per insect and 0.34 × 10(-4) g per egg after the feeding. The expressions of several reproduction-related genes were upregulated, such as the yolk-development-related genes Ovo-781 and vitellogenin (Vg) were increased by 5.33- and 6.77-folds, respectively. This study shows that TiO2 NPs feeding at low concentration can enhance the reproduction of B. mori, and these results are useful in developing new methods to improve fecundity in B. mori and providing new clues for its broad biological applications.

  12. Effect of degumming time on silkworm silk fibre for biodegradable polymer composites

    NASA Astrophysics Data System (ADS)

    Ho, Mei-po; Wang, Hao; Lau, Kin-tak

    2012-02-01

    Recently, many studies have been conducted on exploitation of natural materials for modern product development and bioengineering applications. Apart from plant-based materials (such as sisal, hemp, jute, bamboo and palm fibre), animal-based fibre is a kind of sustainable natural materials for making novel composites. Silkworm silk fibre extracted from cocoon has been well recognized as a promising material for bio-medical engineering applications because of its superior mechanical and bioresorbable properties. However, when producing silk fibre reinforced biodegradable/bioresorbable polymer composites, hydrophilic sericin has been found to cause poor interfacial bonding with most polymers and thus, it results in affecting the resultant properties of the composites. Besides, sericin layers on fibroin surface may also cause an adverse effect towards biocompatibility and hypersensitivity to silk for implant applications. Therefore, a proper pre-treatment should be done for sericin removal. Degumming is a surface modification process which allows a wide control of the silk fibre's properties, making the silk fibre possible to be used for the development and production of novel bio-composites with unique/specific mechanical and biodegradable properties. In this paper, a cleaner and environmentally friendly surface modification technique for tussah silk in polymer based composites is proposed. The effectiveness of different degumming parameters including degumming time and temperature on tussah silk is discussed through the analyses of their mechanical and morphological properties. Based on results obtained, it was found that the mechanical properties of tussah silk are affected by the degumming time due to the change of the fibre structure and fibroin alignment.

  13. Phosphorylation of Ser-204 and Tyr-405 in human malonyl-CoA decarboxylase expressed in silkworm Bombyx mori regulates catalytic decarboxylase activity.

    PubMed

    Hwang, In-Wook; Makishima, Yu; Suzuki, Tomohiro; Kato, Tatsuya; Park, Sungjo; Terzic, Andre; Chung, Shin-Kyo; Park, Enoch Y

    2015-11-01

    Decarboxylation of malonyl-CoA to acetyl-CoA by malonyl-CoA decarboxylase (MCD; EC 4.1.1.9) is a vital catalytic reaction of lipid metabolism. While it is established that phosphorylation of MCD modulates the enzymatic activity, the specific phosphorylation sites associated with the catalytic function have not been documented due to lack of sufficient production of MCD with proper post-translational modifications. Here, we used the silkworm-based Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid system to express human MCD (hMCD) and mapped phosphorylation effects on enzymatic function. Purified MCD from silkworm displayed post-translational phosphorylation and demonstrated coherent enzymatic activity with high yield (-200 μg/silkworm). Point mutations in putative phosphorylation sites, Ser-204 or Tyr-405 of hMCD, identified by bioinformatics and proteomics analyses reduced the catalytic activity, underscoring the functional significance of phosphorylation in modulating decarboxylase-based catalysis. Identified phosphorylated residues are distinct from the decarboxylation catalytic site, implicating a phosphorylation-induced global conformational change of MCD as responsible in altering catalytic function. We conclude that phosphorylation of Ser-204 and Tyr-405 regulates the decarboxylase function of hMCD leveraging the silkworm-based BmNPV bacmid expression system that offers a fail-safe eukaryotic production platform implementing proper post-translational modification such as phosphorylation.

  14. Experimental RNomics and genomic comparative analysis reveal a large group of species-specific small non-message RNAs in the silkworm Bombyx mori

    PubMed Central

    Li, Dandan; Wang, Yanhong; Zhang, Kun; Jiao, Zhujin; Zhu, Xiaopeng; Skogerboe, Geir; Guo, Xiangqian; Chinnusamy, Viswanathan; Bi, Lijun; Huang, Yongping; Dong, Shuanglin; Chen, Runsheng; Kan, Yunchao

    2011-01-01

    Accumulating evidences show that small non-protein coding RNAs (ncRNAs) play important roles in development, stress response and other cellular processes. The silkworm is an important model for studies on insect genetics and control of lepidopterous pests. Here, we have performed the first systematic identification and analysis of intermediate size ncRNAs (50–500 nt) in the silkworm. We identified 189 novel ncRNAs, including 141 snoRNAs, six snRNAs, three tRNAs, one SRP and 38 unclassified ncRNAs. Forty ncRNAs showed significantly altered expression during silkworm development or across specific stage transitions. Genomic comparisons revealed that 123 of these ncRNAs are potentially silkworm-specific. Analysis of the genomic organization of the ncRNA loci showed that 32.62% of the novel snoRNA loci are intergenic, and that all the intronic snoRNAs follow the pattern of one-snoRNA-per-intron. Target site analysis predicted a total of 95 2′-O-methylation and pseudouridylation modification sites of rRNAs, snRNAs and tRNAs. Together, these findings provide new clues for future functional study of ncRNA during insect development and evolution. PMID:21227919

  15. Protective effect of the silkworm protein 30Kc6 on human vascular endothelial cells damaged by oxidized low density lipoprotein (Ox-LDL).

    PubMed

    Yu, Wei; Ying, Huihui; Tong, Fudan; Zhang, Chen; Quan, Yanping; Zhang, Yaozhou

    2013-01-01

    Although the 30K family proteins are important anti-apoptotic molecules in silkworm hemolymph, the underlying mechanism remains to be investigated. This is especially the case in human vascular endothelial cells (HUVECs). In this study, a 30K protein, 30Kc6, was successfully expressed and purified using the Bac-to-Bac baculovirus expression system in silkworm cells. Furthermore, the 30Kc6 expressed in Escherichia coli was used to generate a polyclonal antibody. Western blot analysis revealed that the antibody could react specifically with the purified 30Kc6 expressed in silkworm cells. The In vitro cell apoptosis model of HUVEC that was induced by oxidized low density lipoprotein (Ox-LDL) and in vivo atherosclerosis rabbit model were constructed and were employed to analyze the protective effects of the silkworm protein 30Kc6 on these models. The results demonstrated that the silkworm protein 30Kc6 significantly enhanced the cell viability in HUVEC cells treated with Ox-LDL, decreased the degree of DNA fragmentation and markedly reduced the level of 8-isoprostane. This could be indicative of the silkworm protein 30Kc6 antagonizing the Ox-LDL-induced cell apoptosis by inhibiting the intracellular reactive oxygen species (ROS) generation. Furthermore, Ox-LDL activated the cell mitogen activated protein kinases (MAPK), especially JNK and p38. As demonstrated with Western analysis, 30Kc6 inhibited Ox-LDL-induced cell apoptosis in HUVEC cells by preventing the MAPK signaling pathways. In vivo data have demonstrated that oral feeding of the silkworm protein 30Kc6 dramatically improved the conditions of the atherosclerotic rabbits by decreasing serum levels of total triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and total cholesterol (TC). Furthermore, 30Kc6 alleviated the extent of lesions in aorta and liver in the atherosclerotic rabbits. These data are not only helpful in understanding the anti

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

    SciTech Connect

    Wang Shengpeng; Guo Tingqing; Guo Xiuyang; Huang Junting; Lu Changde . E-mail: cdlu@sibs.ac.cn

    2006-03-24

    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. TAL effectors mediate high-efficiency transposition of the piggyBac transposon in silkworm Bombyx mori L.

    PubMed

    Ye, Lupeng; You, Zhengying; Qian, Qiujie; Zhang, Yuyu; Che, Jiaqian; Song, Jia; Zhong, Boxiong

    2015-11-26

    The piggyBac (PB) transposon is one of the most useful transposable elements, and has been successfully used for genetic manipulation in more than a dozen species. However, the efficiency of PB-mediated transposition is still insufficient for many purposes. Here, we present a strategy to enhance transposition efficiency using a fusion of transcription activator-like effector (TALE) and the PB transposase (PBase). The results demonstrate that the TALE-PBase fusion protein which is engineered in this study can produce a significantly improved stable transposition efficiency of up to 63.9%, which is at least 7 times higher than the current transposition efficiency in silkworm. Moreover, the average number of transgene-positive individuals increased up to 5.7-fold, with each positive brood containing an average of 18.1 transgenic silkworms. Finally, we demonstrate that TALE-PBase fusion-mediated PB transposition presents a new insertional preference compared with original insertional preference. This method shows a great potential and value for insertional therapy of many genetic diseases. In conclusion, this new and powerful transposition technology will efficiently promote genetic manipulation studies in both invertebrates and vertebrates.

  18. [Identification and expression patterns of anterior silk gland specific cuticle protein Bm11721 in the silkworm (Bombyx mori)].

    PubMed

    Xie, Kang; Wang, Xin; Chen, Huifang; Li, Yi; Song, Qianru; Zhao, Ping

    2016-01-01

    The silk gland of silkworm is the organ of silk protein synthesis and secretion. According to the morphological and functional differences, silk gland can be divided into anterior silk gland (ASG), middle silk gland (MSG) and posterior silk gland (PSG). ASG is the place for silk proteins conformation changes although it cannot synthetize silk proteins. ASG has narrow luminal structures and rigid wall which consists of chitin and cuticle proteins so that it can provide the shearing force which plays an important role in the silk protein conformation changes. The objective of this study is to identify the new chitin binding proteins in ASG of silkworm (Bombyx mori), and to analyze their expression patterns in different tissues. We identified a cuticle protein with chitin binding domain Bml1721 (GenBank Accession No. NM-001173285.1) by chitin affinity chromatography column. We also expressed the recombinant protein as inclusion body using the prokaryotic expression system, and then successfully purified the recombinant protein by nickel affinity chromatography column to generate the polyclonal antibodies. The expression patterns analysis in various tissues showed that both in transcriptional and protein levels Bm11721 was specifically expressed in ASG. Furthermore, the expression level of Bm 11721 protein was unchanged during the 5th instar. Immunofluorescence analysis revealed that Bm1 1721 was located in the ASG inner membrane. It is proposed that Bm11721 is a component of inner membrane and probably provides the shearing force for conformational changes.

  19. Impact of Single and Stacked Insect-Resistant Bt-Cotton on the Honey Bee and Silkworm

    PubMed Central

    Niu, Lin; Ma, Yan; Mannakkara, Amani; Zhao, Yao; Ma, Weihua; Lei, Chaoliang; Chen, Lizhen

    2013-01-01

    Transgenic insect-resistant cotton (Bt cotton) has been extensively planted in China, but its effects on non-targeted insect species such as the economically important honey bee (Apis mellifera) and silkworm (Bombyx mori) currently are unknown. In this study, pollen from two Bt cotton cultivars, one expressing Cry1Ac/EPSPS and the other expressing Cry1Ac/Cry2Ab, were used to evaluate the effects of Bt cotton on adult honey bees and silkworm larvae. Laboratory feeding studies showed no adverse effects on the survival, cumulative consumption, and total hemocyte count (THC) of A. mellifera fed with Bt pollen for 7 days. No effects on the survival or development of B. mori larvae were observed either. A marginally significant difference between Cry1Ac/Cry2Ab cotton and the conventional cotton on the THC of the 3rd day of 5th B. mori instar larvae was observed only at the two highest pollen densities (approximately 900 and 8000 grains/cm2), which are much higher than the pollen deposition that occurs under normal field conditions. The results of this study show that pollen of the tested Bt cotton varieties carried no lethal or sublethal risk for A. mellifera, and the risk for B. mori was negligible. PMID:24039838

  20. Ecdysone Receptor (EcR) Is Involved in the Transcription of Cell Cycle Genes in the Silkworm

    PubMed Central

    Qian, Wenliang; Kang, Lixia; Zhang, Tianlei; Meng, Meng; Wang, Yonghu; Li, Zhiqing; Xia, Qingyou; Cheng, Daojun

    2015-01-01

    EcR (ecdysone receptor)-mediated ecdysone signaling pathway contributes to regulate the transcription of genes involved in various processes during insect development. In this work, we detected the expression of EcR gene in silkworm ovary-derived BmN4 cells and found that EcR RNAi result in an alteration of cell shape, indicating that EcR may orchestrate cell cycle progression. EcR RNAi and EcR overexpression analysis revealed that in the cultured BmN4 cells, EcR respectively promoted and suppressed the transcription of E2F-1 and CycE, two genes controlling cell cycle progression. Further examination demonstrated that ecdysone application in BmN4 cells not only changed the transcription of these two cell cycle genes like that under EcR overexpression, but also induced cell cycle arrest at G2/M phase. In vivo analysis confirmed that E2F-1 expression was elevated in silk gland of silkworm larvae after ecdysone application, which is same as its response to ecdysone in BmN4 cells. However, ecdysone also promotes CycE transcription in silk gland, and this is converse with the observation in BmN4 cells. These results provide new insights into understanding the roles of EcR-mediated ecdysone signaling in the regulation of cell cycle. PMID:25654229

  1. Translationally Controlled Tumor Protein, a Dual Functional Protein Involved in the Immune Response of the Silkworm, Bombyx mori

    PubMed Central

    Hua, Xiaoting; Song, Liang; Xia, Qingyou

    2013-01-01

    Insect gut immunity is the first line of defense against oral infection. Although a few immune-related molecules in insect intestine has been identified by genomics or proteomics approach with comparison to well-studied tissues, such as hemolymph or fat body, our knowledge about the molecular mechanism underlying the gut immunity which would involve a variety of unidentified molecules is still limited. To uncover additional molecules that might take part in pathogen recognition, signal transduction or immune regulation in insect intestine, a T7 phage display cDNA library of the silkworm midgut is constructed. By use of different ligands for biopanning, Translationally Controlled Tumor Protein (TCTP) has been selected. BmTCTP is produced in intestinal epithelial cells and released into the gut lumen. The protein level of BmTCTP increases at the early time points during oral microbial infection and declines afterwards. In vitro binding assay confirms its activity as a multi-ligand binding molecule and it can further function as an opsonin that promotes the phagocytosis of microorganisms. Moreover, it can induce the production of anti-microbial peptide via a signaling pathway in which ERK is required and a dynamic tyrosine phosphorylation of certain cytoplasmic membrane protein. Taken together, our results characterize BmTCTP as a dual-functional protein involved in both the cellular and the humoral immune response of the silkworm, Bombyx mori. PMID:23894441

  2. Genome-Wide Identification, Characterization and Expression Analysis of the Solute Carrier 6 Gene Family in Silkworm (Bombyx mori)

    PubMed Central

    Tang, Xin; Liu, Huawei; Chen, Quanmei; Wang, Xin; Xiong, Ying; Zhao, Ping

    2016-01-01

    The solute carrier 6 (SLC6) gene family, initially known as the neurotransmitter transporters, plays vital roles in the regulation of neurotransmitter signaling, nutrient absorption and motor behavior. In this study, a total of 16 candidate genes were identified as SLC6 family gene homologs in the silkworm (Bombyx mori) genome. Spatio-temporal expression patterns of silkworm SLC6 gene transcripts indicated that these genes were highly and specifically expressed in midgut, brain and gonads; moreover, these genes were expressed primarily at the feeding stage or adult stage. Levels of expression for most midgut-specific and midgut-enriched gene transcripts were down-regulated after starvation but up-regulated after re-feeding. In addition, we observed that expression levels of these genes except for BmSLC6-15 and BmGT1 were markedly up-regulated by a juvenile hormone analog. Moreover, brain-enriched genes showed differential expression patterns during wandering and mating processes, suggesting that these genes may be involved in modulating wandering and mating behaviors. Our results improve our understanding of the expression patterns and potential physiological functions of the SLC6 gene family, and provide valuable information for the comprehensive functional analysis of the SLC6 gene family. PMID:27706106

  3. Impact of single and stacked insect-resistant Bt-cotton on the honey bee and silkworm.

    PubMed

    Niu, Lin; Ma, Yan; Mannakkara, Amani; Zhao, Yao; Ma, Weihua; Lei, Chaoliang; Chen, Lizhen

    2013-01-01

    Transgenic insect-resistant cotton (Bt cotton) has been extensively planted in China, but its effects on non-targeted insect species such as the economically important honey bee (Apis mellifera) and silkworm (Bombyx mori) currently are unknown. In this study, pollen from two Bt cotton cultivars, one expressing Cry1Ac/EPSPS and the other expressing Cry1Ac/Cry2Ab, were used to evaluate the effects of Bt cotton on adult honey bees and silkworm larvae. Laboratory feeding studies showed no adverse effects on the survival, cumulative consumption, and total hemocyte count (THC) of A. mellifera fed with Bt pollen for 7 days. No effects on the survival or development of B. mori larvae were observed either. A marginally significant difference between Cry1Ac/Cry2Ab cotton and the conventional cotton on the THC of the 3(rd) day of 5(th) B. mori instar larvae was observed only at the two highest pollen densities (approximately 900 and 8000 grains/cm(2)), which are much higher than the pollen deposition that occurs under normal field conditions. The results of this study show that pollen of the tested Bt cotton varieties carried no lethal or sublethal risk for A. mellifera, and the risk for B. mori was negligible. PMID:24039838

  4. Histomorphometric and transcriptomic features characterize silk glands' development during the molt to intermolt transition process in silkworm.

    PubMed

    Hu, Wenbo; Liu, Chun; Cheng, Tingcai; Li, Wei; Wang, Niannian; Xia, Qingyou

    2016-09-01

    The molt-intermolt cycle is an essential feature in holometabolous and hemimetabolous insects' development. In the silkworm, silk glands are under dramatic morphological and functional changes with fibroin genes' transcription being repeatedly turned off and on during the molt-intermolt cycles. However, the molecular mechanisms controlling it are still unknown. Here, silk gland's histomorphology and transcriptome analysis were used to characterize changes in its structure and gene expression patterns from molt to intermolt stages. By using section staining and transmission electron microscope, a renewable cell damage was detected in the silk gland at the molt stage, and an increased number of autophagosomes and lysosomes were found in silk gland cells' cytoplasm. Next, by using RNA sequencing, 54,578,413 reads were obtained, of which 85% were mapped to the silkworm reference genome. The expression level analysis of silk protein genes and silk gland transcription factors revealed that fibroin heavy chain, fibroin light chain, P25/fhx, sericin1, sericin3 and Dimm had consistent alteration trends in temporal expression. In addition, differentially expressed genes (DEGs) were identified, and most of the DEGs associated with ecdysone signal transduction, mRNA degradation, protein proteolysis, and autophagy were significantly down-regulated in the transition from molt to intermolt, suggesting that these pathways were activated for the silk gland renewal. These findings provide insights into the molecular mechanisms of silk gland development and silk protein genes transcriptional regulation during the molt to intermolt transition process. PMID:27395780

  5. Oral Administration of Silkworm-Produced GAD65 and Insulin Bi-Autoantigens against Type 1 Diabetes

    PubMed Central

    Liu, Baoping; Yue, Yuan; Yang, Yun; Jin, Yongfeng

    2016-01-01

    Induction of mucosal tolerance by oral administration of protein antigens is a potential therapeutic strategy for preventing and treating type 1 diabetes (T1D); however, the requirement for a large dosage of protein limits clinical applications because of the low efficacy. In this study, we generated a fusion protein CTB-Ins-GAD composed of CTB (cholera toxin B subunit), insulin, and three copies of GAD65 peptide 531–545, which were efficiently produced in silkworm pupae, to evaluate its protective effect against T1D. We demonstrate that oral administration of CTB-Ins-GAD suppressed T1D by up to 78%, which is much more effective than GAD65 single-antigen treatment. Strikingly, CTB-Ins-GAD enhance insulin- and GAD65-specific Th2-like immune responses, which repairs the Th1/Th2 imbalance and increases the number of CD4+CD25+Foxp3+ T cell and suppresses insulin- and GAD65-reactive spleen T lymphocyte proliferation and migration. Our results strongly suggest that the combined dual antigens promote the induction of oral tolerance, thus providing an effective and economic immunotherapy against T1D in combination with a silkworm bioreactor. PMID:26783749

  6. Genome-wide identification and expression profiling of the fatty acid desaturase gene family in the silkworm, Bombyx mori.

    PubMed

    Chen, Q M; Cheng, D J; Liu, S P; Ma, Z G; Tan, X; Zhao, P

    2014-01-01

    Fatty acid desaturases exist in all living organisms and play important roles in many different biologic processes, such as fatty acid metabolism, lipid biosynthetic processes, and pheromone biosynthetic processes. Using the available silkworm genome sequence, we identified 14 candidate fatty acid desaturase genes. Eleven genes contain 3 conserved histidine cluster motifs and 4 transmembrane domains, but their N-terminal residues exhibit obvious diversity. Phylogenetic analysis revealed that there are 6 groups; Bmdesat1 and Bmdesat5-8 were clustered into group 2, which is involved in Δ11 desaturation activity, and Bmdesat3-4 were grouped in group 1, which is involved in Δ9 desaturation activity. Twelve of the 14 genes have expressed sequence tag evidence. Microarray data and reverse transcription polymerase chain reaction analysis demonstrated that Bmdesat3-4 and Bmdesat10 were expressed from the larval to moth stages and in multiple tissues on day 3 of 5th instar larvae. Bmdesat9, Bmdesat11, and Bmdesat14 were expressed during the pupal and late-embryonic stage, suggesting that they may take part in fatty acid metabolism to provide energy. These results provide some insights into the functions of individual fatty acid desaturases in silkworm. PMID:24854660

  7. Oral Administration of Silkworm-Produced GAD65 and Insulin Bi-Autoantigens against Type 1 Diabetes.

    PubMed

    Liu, Baoping; Yue, Yuan; Yang, Yun; Jin, Yongfeng

    2016-01-01

    Induction of mucosal tolerance by oral administration of protein antigens is a potential therapeutic strategy for preventing and treating type 1 diabetes (T1D); however, the requirement for a large dosage of protein limits clinical applications because of the low efficacy. In this study, we generated a fusion protein CTB-Ins-GAD composed of CTB (cholera toxin B subunit), insulin, and three copies of GAD65 peptide 531-545, which were efficiently produced in silkworm pupae, to evaluate its protective effect against T1D. We demonstrate that oral administration of CTB-Ins-GAD suppressed T1D by up to 78%, which is much more effective than GAD65 single-antigen treatment. Strikingly, CTB-Ins-GAD enhance insulin- and GAD65-specific Th2-like immune responses, which repairs the Th1/Th2 imbalance and increases the number of CD4(+)CD25(+)Foxp3(+) T cell and suppresses insulin- and GAD65-reactive spleen T lymphocyte proliferation and migration. Our results strongly suggest that the combined dual antigens promote the induction of oral tolerance, thus providing an effective and economic immunotherapy against T1D in combination with a silkworm bioreactor. PMID:26783749

  8. Identification of the silkworm quail gene reveals a crucial role of a receptor guanylyl cyclase in larval pigmentation.

    PubMed

    Yuasa, Masashi; Kiuchi, Takashi; Banno, Yutaka; Katsuma, Susumu; Shimada, Toru

    2016-01-01

    Diverse color patterns on the integument of lepidopteran larvae play important roles in their survival through camouflage, mimicry, sexual signaling, and aposematism. In the silkworm Bombyx mori, many color pattern variations have been preserved in inbred strains making them a good model for elucidating the molecular mechanisms that underlie color pattern formation. In this study, we focused on the silkworm quail (q) mutant, which exhibits abnormalities in multiple pigment biosynthesis pathways. Positional cloning of the q gene revealed that disruption of a guanylyl cyclase gene, BmGC-I, is responsible for its abnormal pigmentation. In q mutants, we identified a 16-bp deletion in the BmGC-I transcript, resulting in the production of a premature stop codon. Knockout of the BmGC-I gene resulted in the q-like abnormal pigmentation, thereby demonstrating that the BmGC-I gene is involved in the pigment biosynthesis pathway in the integument. Moreover, quantitative reverse transcription polymerase chain reaction showed that BmGC-I was strongly expressed in the fourth instar on day 2. Our results suggest that BmGC-I deficiency affects the pigment biosynthesis pathway, which supports the involvement of guanylyl cyclase in larval coloration.

  9. Improved isolation and purification of functional human Fas receptor extracellular domain using baculovirus-silkworm expression system.

    PubMed

    Muraki, Michiro; Honda, Shinya

    2011-11-01

    To achieve an efficient isolation of human Fas receptor extracellular domain (hFasRECD), a fusion protein of hFasRECD with human IgG1 heavy chain Fc domain containing thrombin cleavage sequence at the junction site was overexpressed using baculovirus-silkworm larvae expression system. The hFasRECD part was separated from the fusion protein by the effective cleavage of the recognition site with bovine thrombin. Protein G column treatment of the reaction mixture and the subsequent cation-exchange chromatography provided purified hFasRECD with a final yield of 13.5mg from 25.0 ml silkworm hemolymph. The functional activity of the product was examined by size-exclusion chromatography analysis. The isolated hFasRECD less strongly interacted with human Fas ligand extracellular domain (hFasLECD) than the Fc domain-bridged counterpart, showing the contribution of antibody-like avidity in the latter case. The purified glycosylated hFasRECD presented several discrete bands in the disulphide-bridge non-reducing SDS-PAGE analysis, and virtually all of the components were considered to participate in the binding to hFasLECD. The attached glycans were susceptible to PNGase F digestion, but mostly resistant to Endo Hf digestion under denaturing conditions. One of the components exhibited a higher susceptibility to PNGase F digestion under non-denaturing conditions.

  10. Antiapoptotic activity of 30 kDa lipoproteins family from fat body tissue of silkworm, Bombyx mori.

    PubMed

    Pakkianathan, Britto Cathrin; Singh, Nitin Kumar; König, Simone; Krishnan, Muthukalingan

    2015-10-01

    The family of 30 kDa lipoproteins (LP1-5) is abundant in silkworm pupa fat body (FB) and hemolymph. One of its members, the 29 kDa protein decreased in concentration from peripheral (PP) FB tissue but was sustained in perivisceral (PV) FB tissue at the time of apoptosis. This study investigated the correlation of the 30 kDa proteins with FB apoptosis. Two protein fractions were purified, a 29 and a 30/31 kDa protein fraction, and they were used to test for activity against actinomycin D-induced apoptosis in the FB tissues. Concentrations as little as 50 μg/mL of the 29 kDa protein fraction efficiently inhibited apoptosis. Less antiapoptotic activity was detected for the higher MW fraction; DNA fragmentation was observed in FB tissue treated with 50 μg/mL of the 30/31 kDa fraction. The viability of the cells in the 29 kDa protein-supplemented culture was 40% higher than in the 31 kDa protein-supplemented culture. However, the 30 kDa lipoproteins were not able to prevent scheduled FB degeneration during silkworm metamorphosis. Thus, it is hypothesized that the antiapoptotic 29 kDa protein needs to be proteolytically degraded by a regulatory mechanism to allow programmed cell death of FB tissue.

  11. [Identification and expression patterns of anterior silk gland specific cuticle protein Bm11721 in the silkworm (Bombyx mori)].

    PubMed

    Xie, Kang; Wang, Xin; Chen, Huifang; Li, Yi; Song, Qianru; Zhao, Ping

    2016-01-01

    The silk gland of silkworm is the organ of silk protein synthesis and secretion. According to the morphological and functional differences, silk gland can be divided into anterior silk gland (ASG), middle silk gland (MSG) and posterior silk gland (PSG). ASG is the place for silk proteins conformation changes although it cannot synthetize silk proteins. ASG has narrow luminal structures and rigid wall which consists of chitin and cuticle proteins so that it can provide the shearing force which plays an important role in the silk protein conformation changes. The objective of this study is to identify the new chitin binding proteins in ASG of silkworm (Bombyx mori), and to analyze their expression patterns in different tissues. We identified a cuticle protein with chitin binding domain Bml1721 (GenBank Accession No. NM-001173285.1) by chitin affinity chromatography column. We also expressed the recombinant protein as inclusion body using the prokaryotic expression system, and then successfully purified the recombinant protein by nickel affinity chromatography column to generate the polyclonal antibodies. The expression patterns analysis in various tissues showed that both in transcriptional and protein levels Bm11721 was specifically expressed in ASG. Furthermore, the expression level of Bm 11721 protein was unchanged during the 5th instar. Immunofluorescence analysis revealed that Bm1 1721 was located in the ASG inner membrane. It is proposed that Bm11721 is a component of inner membrane and probably provides the shearing force for conformational changes. PMID:27363199

  12. US Department of Energy Oak Ridge Operations Environmental Management Public Involvement Plan for the Oak Ridge Reservation

    SciTech Connect

    1996-03-01

    This document was prepared in accordance with CERCLA requirements for writing community relations plans. It includes information on how the DOE Oak Ridge Operations Office prepares and executes Environmental Management Community relations activities. It is divided into three sections: the public involvement plan, public involvement in Oak Ridge, and public involvement in 1995. Four appendices are also included: environmental management in Oak Ridge; community and regional overview; key laws, agreements, and policy; and principal contacts.

  13. Field Use of NMIS at Oak Ridge

    SciTech Connect

    Chiang, L.G.; Conger, M.; Hughes, S.S.; Mattingly, J.K.; McEvers, J.A.; Mihalczo, J.T.; Mullens, J.A.; Perez, R.B.; Turner, C.R.; Uckan, T.; Valentine, T.E.

    1999-08-26

    The Nuclear Materials Identification System (NMIS), developed by the Oak Ridge National Laboratory and Oak Ridge Y-12 Plant (Y-12), has been successfully used at Y-12 for nuclear material control and accountability (NMC&A). It is particularly useful in the high gamma-ray background of storage arrays and for shielded HEU. With three systems in use at Y-12, NMIS has enhanced the NMC&A capability for verification and for confirmation of materials in storage and for HEU receipts by providing capability not available or practical by other NDA methods for safeguards. It has recently cost-effectively quantified the HEU mass and enrichment of hundreds of HEU metal items to within a total spread of {+-} 5% (3 sigma) with and mean deviations for all HEU verified of + 0.2% for mass and {minus}0.2% for enrichment. Three cart portable systems are easily moved around with minimal impact on facility operations since no permanent dedicated floor space is required. The positive impact of NMIS at the Oak Ridge Y-12 Plant is improved and more cost effective NMC&A as well as the resolution of NMC&A findings. Its operation at the Y-12 Plant is essential for compliance with the NMC&A requirements of the US Department of Energy. NMIS portability has allowed one system to be moved temporarily to the former K-25 Gaseous Diffusion Plant for characterization of a large deposit of hydrated uranyl fluoride. The impact of this NMIS application was enhanced and verified nuclear criticality safety that led to the safe removal of a large deposit originally estimated by gamma-ray spectrometry and neutron counting to contain 1300 kg of 3.3 wt% {sup 235}U material. NMIS has also been operational at Los Alamos National Laboratory and Pantex.

  14. Comprehensive integrated planning: A process for the Oak Ridge Reservation, Oak Ridge, Tennessee

    SciTech Connect

    1998-05-01

    The Oak Ridge Comprehensive Integrated Plan is intended to assist the US Department of Energy (DOE) and contractor personnel in implementing a comprehensive integrated planning process consistent with DOE Order 430.1, Life Cycle Asset Management and Oak Ridge Operations Order 430. DOE contractors are charged with developing and producing the Comprehensive Integrated Plan, which serves as a summary document, providing information from other planning efforts regarding vision statements, missions, contextual conditions, resources and facilities, decision processes, and stakeholder involvement. The Comprehensive Integrated Plan is a planning reference that identifies primary issues regarding major changes in land and facility use and serves all programs and functions on-site as well as the Oak Ridge Operations Office and DOE Headquarters. The Oak Ridge Reservation is a valuable national resource and is managed on the basis of the principles of ecosystem management and sustainable development and how mission, economic, ecological, social, and cultural factors are used to guide land- and facility-use decisions. The long-term goals of the comprehensive integrated planning process, in priority order, are to support DOE critical missions and to stimulate the economy while maintaining a quality environment.

  15. Sporulation capacity of Phytophthora ramorum on northern red oak and chestnut oak

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Branches from six 2 to 3-year old northern red and chestnut oak seedlings were dip-inoculated with ca. 5,000 sporangia per milliliter of Phytophthora ramorum isolate Pr-6 and incubated at 100 percent relative humidity in dew chambers for 6 days. Three plants were then used to assess sporangia produ...

  16. Infectivity and sporulation of Phytophthora ramorum on northern red oak and chestnut oak

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Branches from northern red and chestnut oak seedlings were dip-inoculated with 5,000 sporangia per milliliter of Phytophthora ramorum and incubated at 100 percent relative humidity in dew chambers for 6 days. Three plants were then used to assess sporangia production, while the other three plants w...

  17. Hormonal regulation and developmental role of Krüppel homolog 1, a repressor of metamorphosis, in the silkworm Bombyx mori.

    PubMed

    Kayukawa, Takumi; Murata, Mika; Kobayashi, Isao; Muramatsu, Daisuke; Okada, Chieko; Uchino, Keiro; Sezutsu, Hideki; Kiuchi, Makoto; Tamura, Toshiki; Hiruma, Kiyoshi; Ishikawa, Yukio; Shinoda, Tetsuro

    2014-04-01

    Juvenile hormone (JH) has an ability to repress the precocious metamorphosis of insects during their larval development. Krüppel homolog 1 (Kr-h1) is an early JH-inducible gene that mediates this action of JH; however, the fine hormonal regulation of Kr-h1 and the molecular mechanism underlying its antimetamorphic effect are little understood. In this study, we attempted to elucidate the hormonal regulation and developmental role of Kr-h1. We found that the expression of Kr-h1 in the epidermis of penultimate-instar larvae of the silkworm Bombyx mori was induced by JH secreted by the corpora allata (CA), whereas the CA were not involved in the transient induction of Kr-h1 at the prepupal stage. Tissue culture experiments suggested that the transient peak of Kr-h1 at the prepupal stage is likely to be induced cooperatively by JH derived from gland(s) other than the CA and the prepupal surge of ecdysteroid, although involvement of unknown factor(s) could not be ruled out. To elucidate the developmental role of Kr-h1, we generated transgenic silkworms overexpressing Kr-h1. The transgenic silkworms grew normally until the spinning stage, but their development was arrested at the prepupal stage. The transgenic silkworms from which the CA were removed in the penultimate instar did not undergo precocious pupation or larval-larval molt but fell into prepupal arrest. This result demonstrated that Kr-h1 is indeed involved in the repression of metamorphosis but that Kr-h1 alone is incapable of implementing normal larval molt. Moreover, the expression profiles and hormonal responses of early ecdysone-inducible genes (E74, E75, and Broad) in transgenic silkworms suggested that Kr-h1 is not involved in the JH-dependent modulation of these genes, which is associated with the control of metamorphosis.

  18. Reducing blood glucose levels in TIDM mice with an orally administered extract of sericin from hIGF-I-transgenic silkworm cocoons.

    PubMed

    Song, Zuowei; Zhang, Mengyao; Xue, Renyu; Cao, Guangli; Gong, Chengliang

    2014-05-01

    In previous studies, we reported that the blood glucose levels of mice with type I diabetes mellitus (TIDM) was reduced with orally administered silk gland powder from silkworms transgenic for human insulin-like growth factor-I (hIGF-I). However, potential safety hazards could not be eliminated because the transgenic silk gland powder contained heterologous DNA, including the green fluorescent protein (gfp) and neomycin resistance (neo) genes. These shortcomings might be overcome if the recombinant hIGF-I were secreted into the sericin layer of the cocoon. In this study, silkworm eggs were transfected with a novel piggyBac transposon vector, pigA3GFP-serHS-hIGF-I-neo, containing the neo, gfp, and hIGF-I genes controlled by the sericin-1 (ser-1) promoter with the signal peptide DNA sequence of the fibrin heavy chain (Fib-H) and a helper plasmid containing the piggyBac transposase sequence under the control of the Bombyx mori actin 3 (A3) promoter, using sperm-mediated gene transfer to generate the transformed silkworms. The hIGF-I content estimated by enzyme-linked immunosorbent assay was approximately 162.7 ng/g. To estimate the biological activity of the expressed hIGF-I, streptozotocin-induced TIDM mice were orally administered sericin from the transgenic silkworm. The blood glucose levels of the mice were significantly reduced, suggesting that the extract from the transgenic hIGF-I silkworm cocoons can be used as an orally administered drug.

  19. Oak Ridge Reservation environmental report for 1991

    SciTech Connect

    Mucke, P.C.

    1992-10-01

    The first two volumes of this report present data and supporting narratives regarding the impact of the US Department of Energy's (DOE's) Oak Ridge Reservation (ORR) on its surrounding environs and the public during 1991. Volume 1 includes all narrative descriptions, summaries, and conclusions and is intended to be a stand-alone'' report for the reader who does not want to review in detail all of the 1991 data for the ORR. This volume, Volume 2, includes the detailed data formats that ensure all the environmental data are represented. Narratives are not included. The information in Vol. 2 is addressed and analyzed in Vol. 1.

  20. Oak Ridge Leadership Computing Facility Position Paper

    SciTech Connect

    Oral, H Sarp; Hill, Jason J; Thach, Kevin G; Podhorszki, Norbert; Klasky, Scott A; Rogers, James H; Shipman, Galen M

    2011-01-01

    This paper discusses the business, administration, reliability, and usability aspects of storage systems at the Oak Ridge Leadership Computing Facility (OLCF). The OLCF has developed key competencies in architecting and administration of large-scale Lustre deployments as well as HPSS archival systems. Additionally as these systems are architected, deployed, and expanded over time reliability and availability factors are a primary driver. This paper focuses on the implementation of the Spider parallel Lustre file system as well as the implementation of the HPSS archive at the OLCF.

  1. Presence of Triploids among Oak Species

    PubMed Central

    Dzialuk, Artur; Chybicki, Igor; Welc, Monika; Śliwińska, Elwira; Burczyk, Jaroslaw

    2007-01-01

    Background and Aims Polyploids, organisms that have multiple sets of chromosomes, are common in certain plant and animal taxa. However, there are only a few reports of intraspecific ploidy variation within the genus Quercus. The aim of the study was to investigate the suspected ploidy level of two oaks that have unusual microsatellite banding patterns. Methods Polyploidy was investigated by using microsatellite analysis, stomata length measurements and nuclear DNA content estimation by flow cytometry. Key Results Each putative triploid tree has patterns of microsatellite variation unexpected for diploid genomes, with up to three alleles at some loci, significantly longer stomata and 1·5 times more DNA per nucleus compared with diploids. Conclusions To our knowledge, this report contains the first evidence for triploidy in Q. petraea and confirmation of this phenomenon in Q. robur. Regardless of the positive or negative aspects of the presence of triploid oaks in forest stands, it is of value to be able to screen for them. This study demonstrates that nuclear microsatellites and estimation of DNA content by flow cytometry can readily be used for this purpose. PMID:17409100

  2. The Oak Ridge Isochronous Cyclotron Refurbishment Project

    SciTech Connect

    Mendez, II, Anthony J; Ball, James B; Dowling, Darryl T; Mosko, Sigmund W; Tatum, B Alan

    2011-01-01

    The Oak Ridge Isochronous Cyclotron (ORIC) has been in operation for nearly fifty years at the Oak Ridge National Laboratory (ORNL). Presently, it serves as the driver accelerator for the ORNL Holifield Radioactive Ion Beam Facility (HRIBF), where radioactive ion beams are produced using the Isotope Separation Online (ISOL) technique for post-acceleration by the 25URC tandem electrostatic accelerator. Operability and reliability of ORIC are critical issues for the success of HRIBF and have presented increasingly difficult operational challenges for the facility in recent years. In February 2010, a trim coil failure rendered ORIC inoperable for several months. This presented HRIBF with the opportunity to undertake various repairs and maintenance upgrades aimed at restoring the full functionality of ORIC and improving the reliability to a level better than what had been typical over the previous decade. In this paper, we present details of these efforts, including the replacement of the entire trim coil set and measurements of their radial field profile. Comparison of measurements and operating tune parameters with setup code predictions will also be presented.

  3. 60 Years of Great Science (Oak Ridge National Laboratory)

    DOE R&D Accomplishments Database

    2003-01-01

    This issue of Oak Ridge National Laboratory Review (vol. 36, issue 1) highlights Oak Ridge National Laboratory's contributions in more than 30 areas of research and related activities during the past 60 years and provides glimpses of current activities that are carrying on this heritage.

  4. Exploring the Taxonomy of Oaks and Related Tree Species

    ERIC Educational Resources Information Center

    McMaster, Robert T.

    2004-01-01

    A lab in Eastern North America conducted a study to determine the taxonomic relationship between deciduous trees and several species of oaks by calculating the similarity index of all species to be studied. The study enabled students to classify the different species of oaks according to their distinct characteristics.

  5. Oak Ridge Reservation Annual Site environmental report summary for 1994

    SciTech Connect

    1995-09-01

    This document presents a summary of the information collected for the Oak Ridge Reservation 1994 site environmental report. Topics discussed include: Oak Ridge Reservation mission; ecology; environmental laws; community participation; environmental restoration; waste management; radiation effects; chemical effects; risk to public; environmental monitoring; and radionuclide migration.

  6. Evaluation of spectral light management on growth of container-grown willow oak, nuttall oak and summer red maple

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plant response to blue, red, gray or black shade cloth was evaluated with willow oak (Quercus phellos L.), Nuttall oak (Quercus nuttallii Palmer, Nuttall) and Summer Red maple (Acer rubrum L. ‘Summer Red’) liners. Light transmitted through the colored shade cloth had no influence on germination of ...

  7. 77 FR 68818 - Notice of Inventory Completion: U.S. Department of Energy Oak Ridge Office, Oak Ridge, TN

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-16

    ... affiliation with the human remains and associated funerary object should contact the U.S. Department of Energy... funerary object in the possession of the U.S. Department of Energy Oak Ridge Office. The human remains and... acquisition of the human remains and associated funerary object, the U.S. Department of Energy Oak...

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

    PubMed Central

    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

  9. Source document for waste area groupings at Oak Ridge National Laboratory, Oak Ridge, Tennessee

    SciTech Connect

    Osborne, P.L.; Kuhaida, A.J., Jr.

    1996-09-01

    This document serves as a source document for Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) and other types of documents developed for and pertaining to Environmental Restoration (ER) Program activities at Oak Ridge National Laboratory (ORNL). It contains descriptions of the (1) regulatory requirements for the ORR ER Program, (2) Oak Ridge Reservation (ORR) ER Program, (3) ORNL site history and characterization, and (4) history and characterization of Waste Area Groupings (WAGS) 1-20. This document was created to save time, effort, and money for persons and organizations drafting documents for the ER Program and to improve consistency in the documents prepared for the program. By eliminating the repetitious use of selected information about the program, this document will help reduce the time and costs associated with producing program documents. By serving as a benchmark for selected information about the ER Program, this reference will help ensure that information presented in future documents is accurate and complete.

  10. Oak Ridge Health Studies phase 1 report, Volume 1: Oak Ridge Phase 1 overview

    SciTech Connect

    Yarbrough, M.I.; Van Cleave, M.L.; Turri, P.; Daniel, J.

    1993-09-01

    In July 1991, the State of Tennessee initiated the Health Studies Agreement with the United States Department of Energy to carry out independent studies of possible adverse health effects in people living in the vicinity of the Oak Ridge Reservation. The health studies focus on those effects that could have resulted or could result from exposures to chemicals and radioactivity released at the Reservation since 1942. The major focus of the first phase was to complete a Dose Reconstruction Feasibility Study. This study was designed to find out if enough data exist about chemical and radionuclide releases from the Oak Ridge Reservation to conduct a second phase. The second phase will lead to estimates of the actual amounts or the ``doses`` of various contaminants received by people as a result of off-site releases. Once the doses of various contaminants have been estimated, scientists and physicians will be better able to evaluate whether adverse health effects could have resulted from the releases.

  11. White Oak Creek embayment sediment retention structure: The Oak Ridge model in action

    SciTech Connect

    Van Hoesen, S.D.; Kimmel, B.L.; Page, D.G.; Hudson, G.R.; Wilkerson, R.B.; Zocolla, M.; Kauschinger, J.L.

    1992-12-01

    White Oak Creek is the major surface-water drainage through the Department of Energy (DOE) Oak Ridge National Laboratory (ORNL). Samples taken from the lower portion of the creek revealed high levels of Cesium-137, and lower levels of Cobalt-60 in near-surface sediment. Other contaminants present in the sediment included: lead, mercury, chromium, and PCBS. In October 1990, DOE, US Environmental Protection Agency (EPA), and Tennessee Department of Environment and Conservation (DEC) agreed to initiate a time-critical removal action in accordance with Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) to prevent transport of the contaminated sediments into the Clinch River system. This paper discusses the environmental, regulatory, design, and construction issues that were encountered in conducting the remediation work.

  12. White Oak Creek embayment sediment retention structure: The Oak Ridge model in action

    SciTech Connect

    Van Hoesen, S.D.; Kimmel, B.L. ); Page, D.G.; Hudson, G.R. ); Wilkerson, R.B. ); Zocolla, M. . Nashville District); Kauschinger, J.L. )

    1992-01-01

    White Oak Creek is the major surface-water drainage through the Department of Energy (DOE) Oak Ridge National Laboratory (ORNL). Samples taken from the lower portion of the creek revealed high levels of Cesium-137, and lower levels of Cobalt-60 in near-surface sediment. Other contaminants present in the sediment included: lead, mercury, chromium, and PCBS. In October 1990, DOE, US Environmental Protection Agency (EPA), and Tennessee Department of Environment and Conservation (DEC) agreed to initiate a time-critical removal action in accordance with Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) to prevent transport of the contaminated sediments into the Clinch River system. This paper discusses the environmental, regulatory, design, and construction issues that were encountered in conducting the remediation work.

  13. Melton Valley Storage Tanks Capacity Increase Project, Oak Ridge National Laboratory, Oak Ridge, Tennessee

    SciTech Connect

    1995-04-01

    The US Department of Energy (DOE) proposes to construct and maintain additional storage capacity at Oak Ridge National Laboratory (ORNL), Oak Ridge, Tennessee, for liquid low-level radioactive waste (LLLW). New capacity would be provided by a facility partitioned into six individual tank vaults containing one 100,000 gallon LLLW storage tank each. The storage tanks would be located within the existing Melton Valley Storage Tank (MVST) facility. This action would require the extension of a potable water line approximately one mile from the High Flux Isotope Reactor (HFIR) area to the proposed site to provide the necessary potable water for the facility including fire protection. Alternatives considered include no-action, cease generation, storage at other ORR storage facilities, source treatment, pretreatment, and storage at other DOE facilities.

  14. Spatial distribution of oak mistletoe as it relates to habits of oak woodland frugivores.

    PubMed

    Wilson, Ethan A; Sullivan, Patrick J; Dickinson, Janis L

    2014-01-01

    This study addresses the underlying spatial distribution of oak mistletoe, Phoradendron villosum, a hemi-parasitic plant that provides a continuous supply of berries for frugivorous birds overwintering the oak savanna habitat of California's outer coast range. As the winter community of birds consuming oak mistletoe varies from group-living territorial species to birds that roam in flocks, we asked if mistletoe volume was spatially autocorrelated at the scale of persistent territories or whether the patterns predicted by long-term territory use by western bluebirds are overcome by seed dispersal by more mobile bird species. The abundance of mistletoe was mapped on trees within a 700 ha study site in Carmel Valley, California. Spatial autocorrelation of mistletoe volume was analyzed using the variogram method and spatial distribution of oak mistletoe trees was analyzed using Ripley's K and O-ring statistics. On a separate set of 45 trees, mistletoe volume was highly correlated with the volume of female, fruit-bearing plants, indicating that overall mistletoe volume is a good predictor of fruit availability. Variogram analysis showed that mistletoe volume was spatially autocorrelated up to approximately 250 m, a distance consistent with persistent territoriality of western bluebirds and philopatry of sons, which often breed next door to their parents and are more likely to remain home when their parents have abundant mistletoe. Using Ripley's K and O-ring analyses, we showed that mistletoe trees were aggregated for distances up to 558 m, but for distances between 558 to 724 m the O-ring analysis deviated from Ripley's K in showing repulsion rather than aggregation. While trees with mistletoe were aggregated at larger distances, mistletoe was spatially correlated at a smaller distance, consistent with what is expected based on persistent group territoriality of western bluebirds in winter and the extreme philopatry of their sons. PMID:25389971

  15. Tiger team assessment of the Oak Ridge Y-12 Plant, Oak Ridge, Tennessee

    SciTech Connect

    none,

    1990-02-01

    This document contains findings identified during the Tiger Team Compliance Assessment of the Department of Energy's (DOE's) Y-12 Plant in Oak Ridge, Tennessee. The Y-12 Plant Tiger Team Compliance Assessment is comprehensive in scope. It covers the Environmental, Safety, and Health (including Occupational Safety and Health Administration (OSHA) compliance), and Management areas and determines the plant's compliance with applicable federal (including DOE), state, and local regulations and requirements. 4 figs., 12 tabs.

  16. Removal action report on the Building 3001 canal at Oak Ridge National Laboratory, Oak Ridge, Tennessee

    SciTech Connect

    1997-05-01

    Oak Ridge National Laboratory (ORNL) is a federal facility managed by Lockheed Martin C, Energy Research, Inc., for the U.S. Department of Energy (DOE). ORNL on the Oak Ridge Reservation in East Tennessee at the Anderson and Roane County lines, approximately 38 km (24 miles) west of Knoxville, Tennessee, and 18 km (11 miles) southwest of downtown Oak Ridge. The Oak Ridge Graphite Reactor and its storage and transfer canal are located in Bldg. 3001 in the approximate center of Waste Area Grouping I in the ORNL main complex. 4:1 The Bldg. 3001 Storage Canal is an L-shaped, underground, reinforced-concrete structure running from the back and below the Graphite Reactor in Bldg. 3001 to a location beneath a hot cell in the adjacent Bldg. 3019. The Graphite Reactor was built in 1943 to produce small quantities of plutonium and was subsequently used to produce other isotopes for medical research before it was finally shut down in 1963. The associated canal was used to transport, under water, spent fuel slugs and other isotopes from the back of the reactor to the adjacent Bldg. 31319 hot cell for further processing. During its operation and years subsequent to operation, the canal`s concrete walls and floor became contaminated with radioisotopes from the water.This report documents the activities involved with replacing the canal water with a solid, controlled, low-strength material (CLSM) in response to a Comprehensive Environmental Response, Compensation, and Liability Act non-time-critical removal action.

  17. Site descriptions of environmental restoration units at Oak Ridge National Laboratory, Oak Ridge, Tennessee

    SciTech Connect

    Kuhaida, A.J. Jr.; Parker, A.F.

    1997-02-01

    This report provides summary information on Oak Ridge National Laboratory (ORNL) Environmental Restoration (ER) sites as listed in the Oak Ridge Reservation Federal Facility Agreement (FFA), dated January 1, 1992, Appendix C. The Oak Ridge National Laboratory was built in 1943 as part of the World War II Manhattan Project. The original mission of ORNL was to produce and chemically separate the first gram-quantities of plutonium as part of the national effort to produce the atomic bomb. The current mission of ORNL is to provide applied research and development in support of the U.S. Department of Energy (DOE) programs in nuclear fusion and fission, energy conservation, fossil fuels, and other energy technologies and to perform basic scientific research in selected areas of the physical, life, and environmental sciences. ER is also tasked with clean up or mitigation of environmental impacts resulting from past waste management practices on portions of the approximately 37,000 acres within the Oak Ridge Reservation (ORR). Other installations located within the ORR are the Gaseous Diffusion Plant (K-25) and the Y-12 plant. The remedial action strategy currently integrates state and federal regulations for efficient compliance and approaches for both investigations and remediation efforts on a Waste Area Grouping (WAG) basis. As defined in the ORR FFA Quarterly Report July - September 1995, a WAG is a grouping of potentially contaminated sites based on drainage area and similar waste characteristics. These contaminated sites are further divided into four categories based on existing information concerning whether the data are generated for scoping or remedial investigation (RI) purposes. These areas are as follows: (1) Operable Units (OU); (2) Characterization Areas (CA); (3) Remedial Site Evaluation (RSE) Areas; and (4) Removal Site Evaluation (RmSE) Areas.

  18. Oak Ridge Reservation environmental report for 1989

    SciTech Connect

    Jacobs, V.A.; Wilson, A.R.

    1990-10-01

    The first two volumes of this report are devoted to a presentation of environmental data and supporting narratives for the US Department of Energy's (DOE's) Oak Ridge Reservation (ORR) and surrounding environs during 1989. Volume 1 includes all narrative descriptions, summaries, and conclusions and is intended to be a stand-alone'' report for the ORR for the reader who does not want to review in detail all of the 1989 data. Volume 2 includes the detailed data summarized in a format to ensure that all environmental data are represented in the tables. Narratives are not included in Vol. 2. The tables in Vol. 2 are addressed in Vol. 1. For this reason, Vol. 2 cannot be considered a stand-alone report but is intended to be used in conjunction with Vol. 1. 16 figs., 194 tabs.

  19. Oak Ridge Reservation Environmental report for 1990

    SciTech Connect

    Wilson, A.R.

    1991-09-01

    The first two volumes of this report are devoted to a presentation of environmental data and supporting narratives for the US Department of Energy's (DOE's) Oak Ridge Reservation (ORR) and surrounding environs during 1990. Volume 1 includes all narrative descriptions, summaries, and conclusions and is intended to be a stand-alone'' report for the ORR for the reader who does not want to review in detail all of the 1990 data. Volume 2 includes the detailed data summarized in a format to ensure that all environmental data are represented in the tables. Narratives are not included in Vol. 2. The tables in Vol. 2 are addressed in Vol. 1. For this reason, Vol. 2 cannot be considered a stand-alone report but is intended to be used in conjunction with Vol. 1.

  20. Oak Ridge Reservation environmental report for 1988

    SciTech Connect

    Prince, N.L.

    1989-05-01

    The first two volumes of this report are devoted to a presentation of environmental data and supporting narratives for the US Department of Energy's (DOE's) Oak Ridge Reservation (ORR) and surrounding environs during 1988. Volume 1 includes all narrative descriptions, summaries, and conclusions and is intended to be a stand-alone'' report for the ORR for the reader who does not want to review in detail all of the 1988 data. Volume 2 includes the detailed data summarized in a format to ensure that all environmental data are represented in the tables. Narratives are not included in Volume 2. The tables in Volume 2 are addressed in Volume 1. For this reason, Vol. 2 cannot be considered a stand-alone report but is intended to be used in conjunction with Volume 1.

  1. Site descriptions of environmental restoration units at the Oak Ridge K-25 Site, Oak Ridge, Tennessee

    SciTech Connect

    Goddard, P.L.; Legeay, A.J.; Pesce, D.S.; Stanley, A.M.

    1995-11-01

    This report, Site Descriptions of Environmental Restoration Units at the Oak Ridge K-25 Site, Oak Ridge, Tennessee, is being prepared to assimilate information on sites included in the Environmental Restoration (ER) Program of the K-25 Site, one of three major installations on the Oak Ridge Reservation (ORR) built during World War III as part of the Manhattan Project. The information included in this report will be used to establish program priorities so that resources allotted to the K-25 ER Program can be best used to decrease any risk to humans or the environment, and to determine the sequence in which any remedial activities should be conducted. This document will be updated periodically in both paper and Internet versions. Units within this report are described in individual data sheets arranged alphanumerically. Each data sheet includes entries on project status, unit location, dimensions and capacity, dates operated, present function, lifecycle operation, waste characteristics, site status, media of concern, comments, and references. Each data sheet is accompanied by a photograph of the unit, and each unit is located on one of 13 area maps. These areas, along with the sub-area, unit, and sub-unit breakdowns within them, are outlined in Appendix A. Appendix B is a summary of information on remote aerial sensing and its applicability to the ER program.

  2. The ecdysteroidogenic P450 Cyp302a1/disembodied from the silkworm, Bombyx mori, is transcriptionally regulated by prothoracicotropic hormone.

    PubMed

    Niwa, R; Sakudoh, T; Namiki, T; Saida, K; Fujimoto, Y; Kataoka, H

    2005-10-01

    During larval and pupal development of insects, ecdysone is synthesized in the prothoracic gland (PG). Although several Drosophila genes, including Halloween P450 genes, are known to be important for ecdysteroidogenesis in PG, little is known of the ecdysteroidogenic genes in other insects. Here we report on Cyp302a1/disembodied (dib-Bm), one of the Halloween P450s in the silkworm Bombyx mori that is a carbon-22 hydroxylase. dib-Bm is predominantly expressed in PG and its developmental expression profile is correlated with a change in the ecdysteroid titre in the haemolymph. Furthermore, dib-Bm expression in cultured PGs is significantly induced by treatment with prothoracicotropic hormone. This is the first report on the transcriptional induction of a steroidogenic gene by the tropic hormone in insects. PMID:16164612

  3. Composite Membranes of Recombinant Silkworm Antimicrobial Peptide and Poly (L-lactic Acid) (PLLA) for biomedical application.

    PubMed

    Li, Zhi; Liu, Xuan; Li, Yi; Lan, Xiqian; Leung, Polly Hangmei; Li, Jiashen; Li, Gang; Xie, Maobin; Han, Yanxia; Lin, Xiaofen

    2016-01-01

    Antimicrobial peptides, produced by innate immune system of hosts in response to invading pathogens, are capable of fighting against a spectrum of bacteria, viruses, fungi, parasites and cancer cells. Here, a recombinant silkworm AMP Bmattacin2 from heterologous expression is studied, indicating a broad spectrum of antibacterial activity and showing selective killing ability towards skin and colon cancer cells over their normal cell counterparts. For the purpose of biomedical application, the electrospinning fabrication technique is employed to load Bmattacin2 into PLLA nanofibrous membrane. In addition to a good compatibility with the normal cells, Bmattacin2 loaded nanofibrous membranes demonstrate instant antibacterial effects and sustained anticancer effects. The cancer cell and bacteria targeting dynamics of recombinant Bmattacin2 are investigated. With these characteristics, PLLA/Bmattacin2 composite membranes have a great potential for developing novel biomedical applications such as cancer therapies and wound healing treatments. PMID:27503270

  4. Composite Membranes of Recombinant Silkworm Antimicrobial Peptide and Poly (L-lactic Acid) (PLLA) for biomedical application

    PubMed Central

    Li, Zhi; Liu, Xuan; Li, Yi; Lan, Xiqian; Leung, Polly Hangmei; Li, Jiashen; Li, Gang; Xie, Maobin; Han, Yanxia; Lin, Xiaofen

    2016-01-01

    Antimicrobial peptides, produced by innate immune system of hosts in response to invading pathogens, are capable of fighting against a spectrum of bacteria, viruses, fungi, parasites and cancer cells. Here, a recombinant silkworm AMP Bmattacin2 from heterologous expression is studied, indicating a broad spectrum of antibacterial activity and showing selective killing ability towards skin and colon cancer cells over their normal cell counterparts. For the purpose of biomedical application, the electrospinning fabrication technique is employed to load Bmattacin2 into PLLA nanofibrous membrane. In addition to a good compatibility with the normal cells, Bmattacin2 loaded nanofibrous membranes demonstrate instant antibacterial effects and sustained anticancer effects. The cancer cell and bacteria targeting dynamics of recombinant Bmattacin2 are investigated. With these characteristics, PLLA/Bmattacin2 composite membranes have a great potential for developing novel biomedical applications such as cancer therapies and wound healing treatments. PMID:27503270

  5. The ecdysteroidogenic P450 Cyp302a1/disembodied from the silkworm, Bombyx mori, is transcriptionally regulated by prothoracicotropic hormone.

    PubMed

    Niwa, R; Sakudoh, T; Namiki, T; Saida, K; Fujimoto, Y; Kataoka, H

    2005-10-01

    During larval and pupal development of insects, ecdysone is synthesized in the prothoracic gland (PG). Although several Drosophila genes, including Halloween P450 genes, are known to be important for ecdysteroidogenesis in PG, little is known of the ecdysteroidogenic genes in other insects. Here we report on Cyp302a1/disembodied (dib-Bm), one of the Halloween P450s in the silkworm Bombyx mori that is a carbon-22 hydroxylase. dib-Bm is predominantly expressed in PG and its developmental expression profile is correlated with a change in the ecdysteroid titre in the haemolymph. Furthermore, dib-Bm expression in cultured PGs is significantly induced by treatment with prothoracicotropic hormone. This is the first report on the transcriptional induction of a steroidogenic gene by the tropic hormone in insects.

  6. Beneficial effect of silkworm hemolymph on a CHO cell system: Inhibition of apoptosis and increase of EPO production.

    PubMed

    Choi, Shin Sik; Rhee, Won Jong; Park, Tai Hyun

    2005-09-30

    To produce erythropoietin (EPO), Chinese hamster ovary (CHO) cells were first cultured in a medium containing FBS (growth medium) and then in a serum-free medium containing sodium butyrate (production medium). Sodium butyrate increases recombinant protein production, but also induces apoptosis, which reduces cell viability and productivity. In a previous study, we found that silkworm hemolymph (SH), an insect serum, inhibits the apoptosis of insect and mammalian cells. To overcome sodium butyrate-induced apoptosis, we added SH to growth medium. This pretreatment with SH inhibited the sodium butyrate-induced apoptosis of CHO cells and consequently increased their longevity and their ability to produce EPO. As a result, the volumetric productivity of EPO was increased five-fold. SH was found to inhibit cytochrome c release from mitochondria into the cytosol, and prevented the activation of caspase-3 and other subsequent caspase reactions.

  7. Role of the silkworm argonaute2 homolog gene in double-strand break repair of extrachromosomal DNA.

    PubMed

    Tsukioka, Haruna; Takahashi, Masateru; Mon, Hiroaki; Okano, Kazuhiro; Mita, Kazuei; Shimada, Toru; Lee, Jae Man; Kawaguchi, Yutaka; Koga, Katsumi; Kusakabe, Takahiro

    2006-01-01

    The argonaute protein family provides central components for RNA interference (RNAi) and related phenomena in a wide variety of organisms. Here, we isolated, from a Bombyx mori cell, a cDNA clone named BmAGO2, which is homologous to Drosophila ARGONAUTE2, the gene encoding a repressive factor for the recombination repair of extrachromosomal double-strand breaks (DSBs). RNAi-mediated silencing of the BmAGO2 sequence markedly increased homologous recombination (HR) repair of DSBs in episomal DNA, but had no effect on that in chromosomes. Moreover, we found that RNAi for BmAGO2 enhanced the integration of linearized DNA into a silkworm chromosome via HR. These results suggested that BmAgo2 protein plays an indispensable role in the repression of extrachromosomal DSB repair.

  8. The silkworm glutathione S-transferase gene noppera-bo is required for ecdysteroid biosynthesis and larval development.

    PubMed

    Enya, Sora; Daimon, Takaaki; Igarashi, Fumihiko; Kataoka, Hiroshi; Uchibori, Miwa; Sezutsu, Hideki; Shinoda, Tetsuro; Niwa, Ryusuke

    2015-06-01

    Insect molting and metamorphosis are tightly controlled by ecdysteroids, which are important steroid hormones that are synthesized from dietary sterols in the prothoracic gland. One of the ecdysteroidogenic genes in the fruit fly Drosophila melanogaster is noppera-bo (nobo), also known as GSTe14, which encodes a member of the epsilon class of glutathione S-transferases. In D. melanogaster, nobo plays a crucial role in utilizing cholesterol via regulating its transport and/or metabolism in the prothoracic gland. However, it is still not known whether the orthologs of nobo from other insects are also involved in ecdysteroid biosynthesis via cholesterol transport and/or metabolism in the prothoracic gland. Here we report genetic evidence showing that the silkworm Bombyx mori ortholog of nobo (nobo-Bm; GSTe7) is essential for silkworm development. nobo-Bm is predominantly expressed in the prothoracic gland. To assess the functional importance of nobo-Bm, we generated a B. mori genetic mutant of nobo-Bm using TALEN-mediated genome editing. We show that loss of nobo-Bm function causes larval arrest and a glossy cuticle phenotype, which are rescued by the application of 20-hydroxyecdysone. Moreover, the prothoracic gland cells isolated from the nobo-Bm mutant exhibit an abnormal accumulation of 7-dehydrocholesterol, a cholesterol metabolite. These results suggest that the nobo family of glutathione S-transferases is essential for development and for the regulation of sterol utilization in the prothoracic gland in not only the Diptera but also the Lepidoptera. On the other hand, loss of nobo function mutants of D. melanogaster and B. mori abnormally accumulates different sterols, implying that the sterol utilization in the PG is somewhat different between these two insect species. PMID:25881968

  9. APA-style human milk fat analogue from silkworm pupae oil: Enzymatic production and improving storage stability using alkyl caffeates

    PubMed Central

    Liu, Xi; Wang, Xudong; Pang, Na; Zhu, Weijie; Zhao, Xingyu; Wang, Fangqin; Wu, Fuan; Wang, Jun

    2015-01-01

    Silkworm pupae oil derived from reeling waste is a rich source of α-linolenic acid (ALA), which has multipal applications. ALAs were added in sn-1, 3 positions in a triacylglycerol (TAG) to produce an APA-human milk fat analogues (APA-HMFAs, A: α-linolenic acid, P: palmitic acid). The optimum condition is that tripalmitin to free fatty acids of 1:12 (mole ratio) at 65 °C for 48 h using lipase Lipozyme RM IM. Results show that, the major TAG species that comprised APA-HMFAs were rich in ALA and palmitic acid, which contained 64.52% total unsaturated fatty acids (UFAs) and 97.05% PA at the sn-2 position. The melting point of APA was −27.5 °C which is much lower than tripalmitin (40.5 °C) indicating more plastic character. In addition, the practical application of alkyl caffeates as liposoluble antioxidants in APA was developed. Alkyl caffeate showed a superior IC50 (1.25–1.66 μg/mL) compared to butyl hydroxy anisd (1.67 μg/mL) and L-ascorbic acid-6-palmitate (L-AP) (1.87 μg/mL) in DPPH analysis. The addition of ethyl caffeate to oil achieved a higher UFAs content (73.58%) at high temperatures. Overall, APA was obtained from silkworm pupae oil successfully, and the addition of caffeates extended storage ranges for APA-HMFAs. PMID:26643045

  10. APA-style human milk fat analogue from silkworm pupae oil: Enzymatic production and improving storage stability using alkyl caffeates.

    PubMed

    Liu, Xi; Wang, Xudong; Pang, Na; Zhu, Weijie; Zhao, Xingyu; Wang, Fangqin; Wu, Fuan; Wang, Jun

    2015-12-08

    Silkworm pupae oil derived from reeling waste is a rich source of α-linolenic acid (ALA), which has multipal applications. ALAs were added in sn-1, 3 positions in a triacylglycerol (TAG) to produce an APA-human milk fat analogues (APA-HMFAs, A: α-linolenic acid, P: palmitic acid). The optimum condition is that tripalmitin to free fatty acids of 1:12 (mole ratio) at 65 °C for 48 h using lipase Lipozyme RM IM. Results show that, the major TAG species that comprised APA-HMFAs were rich in ALA and palmitic acid, which contained 64.52% total unsaturated fatty acids (UFAs) and 97.05% PA at the sn-2 position. The melting point of APA was -27.5 °C which is much lower than tripalmitin (40.5 °C) indicating more plastic character. In addition, the practical application of alkyl caffeates as liposoluble antioxidants in APA was developed. Alkyl caffeate showed a superior IC50 (1.25-1.66 μg/mL) compared to butyl hydroxy anisd (1.67 μg/mL) and L-ascorbic acid-6-palmitate (L-AP) (1.87 μg/mL) in DPPH analysis. The addition of ethyl caffeate to oil achieved a higher UFAs content (73.58%) at high temperatures. Overall, APA was obtained from silkworm pupae oil successfully, and the addition of caffeates extended storage ranges for APA-HMFAs.

  11. RNAi KNOCKDOWN OF BmRab3 LED TO LARVA AND PUPA LETHALITY IN SILKWORM Bombyx mori L.

    PubMed

    Singh, Chabungbam Orville; Xin, Hu-hu; Chen, Rui-ting; Wang, Mei-xian; Liang, Shuang; Lu, Yan; Cai, Zi-zheng; Zhang, Deng-pan; Miao, Yun-gen

    2015-06-01

    Rab3 GTPases are known to play key a role in vesicular trafficking, and express highest in brain and endocrine tissues. In mammals, Rab3 GTPases are paralogs unlike in insect. In this study, we cloned Rab3 from the silk gland tissue of silkworm Bombyx mori, and identified it as BmRab3. Our in silico analysis indicated that BmRab3 is an isoform with a theoretical isoelectric point and molecular weight of 5.52 and 24.3 kDa, respectively. Further, BmRab3 showed the C-terminal hypervariability for GGT2 site but having two other putative guanine nucleotide exchange factor/GDP dissociation inhibitor interaction sites. Multiple alignment sequence indicated high similarities of BmRab3 with Rab3 isoforms of other species. The phylogeny tree showed BmRab3 clustered between the species of Tribolium castaneum and Aedes aegypti. Meanwhile, the expression analysis of BmRab3 showed the highest expression in middle silk glands (MSGs) than all other tissues in the third day of fifth-instar larva. Simultaneously, we showed the differential expression of BmRab3 in the early instar larva development, followed by higher expression in male than female pupae. In vivo dsRNA interference of BmRab3 reduced the expression of BmRab3 by 75% compared to the control in the MSGs in the first day. But as the worm grew to the third day, the difference of BmRab3 between knockdown and control was only about 10%. The knockdown later witnessed underdevelopment of the larvae and pharate pupae lethality in the overall development of silkworm B. mori L.

  12. Cloning and expression characteristics of the notch-associated gene BmE(spl)mγ from silkworm, Bombyx mori.

    PubMed

    Liu, Min; Wang, Chan; Li, Dan; Liu, Yue; Sheng, Qing; Lv, Zhengbing; Yu, Wei; Wang, Dan; Zhang, Yaozhou; Nie, Zuoming

    2014-08-01

    The E(spl)mγ gene in Drosophila is a regulatory target gene downstream of the Notch pathway. BmE(spl)mγ (Bombyx mori, E(spl)mγ) is an ortholog of the Drosophila E(spl)mγ gene, and the gene encodes a protein with 248 amino acid residues. This gene was cloned and overexpressed in Escherichia coli BL21(DE3). The recombinant protein was purified and subsequently used to generate a rabbit polyclonal antibody. Western blotting analyses showed that BmE(spl)mγ expression is high in pupa and egg, and low in larva and moth. In the fifth instar larva, the protein levels are high in head, epidermis, sexual gland, trachea, and the fatbody and low in the Malpighian tubule, hemolymph, gut, and silk gland. The further immunohistochemical analyses also showed higher BmE(spl)mγ expression in the head of fifth instar larva and pupa. Of the four moth parts studied, the thorax had the highest expression level. Thus, BmE(spl)mγ might be associated with neurogenesis in silkworm. Furthermore, DAPT (a γ-secretase inhibitor and an indirect inhibitor of Notch) blocking experiments showed that higher concentrations of the blocking agent and a longer processing time reduce the transcription levels of the BmE(spl)mγ gene, demonstrating that the silkworm BmE(spl)mγ gene is associated with the Notch signal pathway. These findings suggest that the function of BmE(spl)mγ may be similar to that of its Drosophila homolog.

  13. Genetic Analysis of the Electrophysiological Response to Salicin, a Bitter Substance, in a Polyphagous Strain of the Silkworm Bombyx mori

    PubMed Central

    Iizuka, Tetsuya; Tamura, Toshiki; Sezutsu, Hideki; Mase, Keisuke; Okada, Eiji; Asaoka, Kiyoshi

    2012-01-01

    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 F1 generation of the two strains had characteristics similar to the Daiankyo strain, consistent with the idea that pph is recessive. In the BF1 progeny between F1 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 BF1 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. PMID:22649537

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

    PubMed

    Iizuka, Tetsuya; Tamura, Toshiki; Sezutsu, Hideki; Mase, Keisuke; Okada, Eiji; Asaoka, Kiyoshi

    2012-01-01

    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. PMID:22649537

  15. Oak Grove Fork Habitat Improvement Project, 1988 Annual Report.

    SciTech Connect

    Bettin, Scott

    1989-04-01

    The Lower Oak Grove Fork of the Clackamas River is a fifth-order tributary of the Clackamas River drainage supporting depressed runs of coho and chinook salmon, and summer and winter steelhead. Habitat condition rating for the Lower Oak Grove is good, but smelt production estimates are below the average for Clackamas River tributaries. Limiting factors in the 3.8 miles of the Lower Oak Grove supporting anadromous fish include an overall lack of quality spawning and rearing habitat. Beginning in 1986. measures to improve fish habitat in the Lower Oak Grove were developed in coordination with the Oregon Department of Fish and Wildlife (ODF&W) and Portland General Electric (PGE) fisheries biologists. Prior to 1986, no measures had been applied to the stream to mitigate for PGE's storage and regulation of flows in the Oak Grove Fork (Timothy Lake, Harriet Lake). Catchable rainbow trout are stocked by ODF&W two or three times a year during the trout fishing season in the lowermost portion of the Oak Grove Fork near two Forest Service campgrounds (Ripplebrook and Rainbow). The 1987 field season marked the third year of efforts to improve fish habitat of the Lower Oak Grove Fork and restore anadromous fish production. The efforts included the development of an implementation plan for habitat improvement activities in the Lower Oak Grove Fork. post-project monitoring. and maintenance of the 1986 improvement structures. No new structures were constructed or placed in 1987. Fiscal year 1988 brought a multitude of changes which delayed implementation of plans developed in 1987. The most prominent change was the withdrawal of the proposed Spotted Owl Habitat Area (SOHA) which overlapped the Oak Grove project implementation area. Another was the change in the Forest Service biologist responsible for implementation and design of this project.

  16. Oak Ridge Dose Reconstruction Project Summary Report; Reports of the Oak Ridge Dose Reconstruction, Vol. 7

    SciTech Connect

    Thomas E. Widner; et. al.

    1999-07-01

    In the early 1990s, concern about the Oak Ridge Reservation's past releases of contaminants to the environment prompted Tennessee's public health officials to pursue an in-depth study of potential off-site health effects at Oak Ridge. This study, the Oak Ridge dose reconstruction, was supported by an agreement between the U.S. Department of Energy (DOE) and the State of Tennessee, and was overseen by a 12-member panel of individuals appointed by Tennessee's Commissioner of Health. The panel requested that the principal investigator for the project prepare the following report, ''Oak Ridge Dose Reconstruction Project Summary Report,'' to serve the following purposes: (1) summarize in a single, less technical report, the methods and results of the various investigations that comprised the Phase II of the dose reconstruction; (2) describe the systematic searching of classified and unclassified historical records that was a vital component of the project; and (3) summarize the less detailed, screening-level assessments that were performed to evaluate the potential health significance of a number of materials, such a uranium, whose priority did not require a complete dose reconstruction effort. This report describes each major step of the dose reconstruction study: (1) the review of thousands of historical records to obtain information relating to past operations at each facility; (2) estimation of the quantity and timing of releases of radioiodines from X-10, of mercury from Y-12, of PCB's from all facilities, and of cesium-137 and other radionuclides from White Oak Creek; (3) evaluation of the routes taken by these contaminants through the environment to nearby populations; and (4) estimation of doses and health risks to exposed groups. Calculations found the highest excess cancer risks for a female born in 1952 who drank goat milk; the highest non-cancer health risk was for children in a farm family exposed to PCBs in and near East Fork Poplar Creek. More detailed

  17. 75 FR 82001 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-29

    ... Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation AGENCY: Department of Energy... Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act... 12, 2011, 6 p.m. ADDRESSES: DOE Information Center, 475 Oak Ridge Turnpike, Oak Ridge,...

  18. 77 FR 45345 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-31

    ... Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation AGENCY: Department of Energy... Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub... Oak Ridge Operations Office, P.O. Box 2001, EM-90, Oak Ridge, TN 37831. Phone (865) 241-3315; Fax...

  19. 76 FR 1415 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-10

    ... Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation AGENCY: Department of Energy... January 12, 2011, ] meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak... Coordinator, Department of Energy Oak Ridge Operations Office, P.O. Box 2001, EM-90, Oak Ridge, TN...

  20. 75 FR 43518 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-26

    ... Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation AGENCY: Department of Energy... Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub..., Department of Energy Oak Ridge Operations Office, P.O. Box 2001, EM-90, Oak Ridge, TN 37831. Phone (865)...

  1. Technical background information for the environmental and safety report, Volume 4: White Oak Lake and Dam

    SciTech Connect

    Oakes, T.W.; Kelly, B.A.; Ohnesorge, W.F.; Eldridge, J.S.; Bird, J.C.; Shank, K.E.; Tsakeres, F.S.

    1982-03-01

    This report has been prepared to provide background information on White Oak Lake for the Oak Ridge National Laboratory Environmental and Safety Report. The paper presents the history of White Oak Dam and Lake and describes the hydrological conditions of the White Oak Creek watershed. Past and present sediment and water data are included; pathway analyses are described in detail.

  2. Screening of contaminants in Waste Area Grouping 2 at Oak Ridge National Laboratory, Oak Ridge, Tennessee. Environmental Restoration Program

    SciTech Connect

    Blaylock, B.G.; Frank, M.L.; Hoffman, F.O.; Hook, L.A.; Suter, G.W.; Watts, J.A.

    1992-07-01

    Waste Area Grouping 2 (WAG 2) of the Oak Ridge National Laboratory (ORNL) is located in the White Oak Creek Watershed and is composed of White Oak Creek Embayment, White Oak Lake and associated floodplain, and portions of White Oak Creek (WOC) and Melton Branch downstream of ORNL facilities. Contaminants leaving other ORNL WAGs in the WOC watershed pass through WAG 2 before entering the Clinch River. Health and ecological risk screening analyses were conducted on contaminants in WAG 2 to determine which contaminants were of concern and would require immediate consideration for remedial action and which contaminants could be assigned a low priority or further study. For screening purposes, WAG 2 was divided into four geographic reaches: Reach 1, a portion of WOC; Reach 2, Melton Branch; Reach 3, White Oak Lake and the floodplain area to the weirs on WOC and Melton Branch; and Reach 4, the White Oak Creek Embayment, for which an independent screening analysis has been completed. Screening analyses were conducted using data bases compiled from existing data on carcinogenic and noncarcinogenic contaminants, which included organics, inorganics, and radionuclides. Contaminants for which at least one ample had a concentration above the level of detection were placed in a detectable contaminants data base. Those contaminants for which all samples were below the level of detection were placed in a nondetectable contaminants data base.

  3. A novel angiotensin-І converting enzyme (ACE) inhibitory peptide from gastrointestinal protease hydrolysate of silkworm pupa (Bombyx mori) protein: Biochemical characterization and molecular docking study.

    PubMed

    Wu, Qiongying; Jia, Junqiang; Yan, Hui; Du, Jinjuan; Gui, Zhongzheng

    2015-06-01

    Silkworm pupa (Bombyx mori) protein was hydrolyzed using gastrointestinal endopeptidases (pepsin, trypsin and α-chymotrypsin). Then, the hydrolysate was purified sequentially by ultrafiltration, gel filtration chromatography and RP-HPLC. A novel ACE inhibitory peptide, Ala-Ser-Leu, with the IC50 value of 102.15μM, was identified by IT-MS/MS. This is the first report of Ala-Ser-Leu from natural protein. Lineweaver-Burk plots suggest that the peptide is a competitive inhibitor against ACE. The molecular docking studies revealed that the ACE inhibition of Ala-Ser-Leu is mainly attributed to forming very strong hydrogen bonds with the S1 pocket (Ala354) and the S2 pocket (Gln281 and His353). The results indicate that silkworm pupa (B. mori) protein or its gastrointestinal protease hydrolysate could be used as a functional ingredient in auxiliary therapeutic foods against hypertension.

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

    PubMed

    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

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

    PubMed

    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

  6. [Cloning and expression profile of Bmyan in the silkworm (Bombyx mori) and experimental validation as one target of microRNA 7].

    PubMed

    Liu, Shiping; Huang, Yaxi; Yin, Jiyun; Wu, Xiaoyan; Zhou, Lanting; Wang, Wei; Xia, Qingyou

    2015-11-01

    microRNAs (miRNAs) are an extensive class of -22-nucleotide (nt) endogenous noncoding RNAs regulating life activities ofmetazoans through binding to 3'-untranslated regions (3'-UTRs) of their target genes. This work aimed to identify yan gene in the silkworm, reveal its expression profile and confirm if it is one target of bmo-miR-7 and, as such, have potential for contributing to better understanding of the molecular mechanisms involved in the metamorphosis of silkworm. Based on homolog searching and PCR amplification, we cloned the coding sequence (CDS) of Bmyan, which encodes 476 amino acid residues and contains SAM-PNT and ETs domains. Quantitative PCR (q-PCR), RT-PCR and microarray data revealed high expression of Bmyan in the head, body wall and ovary of day-3 fifth instar larval silkworm, low or no expression in other tissues. It was lowly expressed in the early larval stages, but highly expressed from late spinning to day 4 pupa. The 3'-UTR of Bmyan was obtained by rapid-amplification of cDNA ends (3'RACE) and predicted to contain two potential recognition sites of bmo-miR-7. The luciferase reporter vector containing the 3'-UTR of Bmyan was constructed and co-transfected into BmE cell line with the mimic of bmo-miR-7 and the decreased relative activity of luciferase showed that Bmyan is one target of bmo-miR-7. This work helps further functional analysis of bmo-miR-7 and Bmyan in the silkworm.

  7. Expression and purification of virus like particles (VLPs) of foot-and-mouth disease virus in Eri silkworm (Samia cynthia ricini) larvae.

    PubMed

    Kumar, Manoj; Saravanan, P; Jalali, S K

    2016-03-01

    Foot-and-mouth disease (FMD) is a highly contagious viral disease, which causes severe economic loss to livestock. Virus like particles (VLPs) produced by recombinant DNA technology are gaining importance because of their immunogenic properties and safety in developing a new vaccine for FMD. In the present study, a practical and economically feasible approach of expression, purification and characterization of VLPs of FMDV in Eri silkworm (Samia cynthia ricini) larvae was described. Although three lepidopteran insect larvae (Helicoverpa armigera, Spodoptera litura and Samia cynthia ricini) were tested for production of VLPs, expression was obtained only in Eri silkworm larvae. High titred recombinant baculovirus encoding the polyprotein P1-2A-3C of FMDV was prepared in Sf9 cells. Injection of recombinant baculovirus into hemocoel of Eri silkworm larvae resulted in increasing levels of expression of VLPs in the hemolymph from 3 to 7 days post infection (dpi) compared to low level expression by oral feeding. The VLPs reacted in Sandwich ELISA with serum raised against whole virus particles of FMDV type O/IND/R2/75 and protein banding pattern of 26, 37 and 47 kDa in Western blotting demonstrated their antigenic resemblance to native virus. Sucrose density gradient purified VLPs were used for immunization of rabbits and guinea pigs for assessing immunogenicity. Further, the reactivity of serum samples of rabbits and guinea pigs in Indirect-ELISA with titres (1.30-2.81 Log10) indicated that the VLPs were antigenic and immunogenic in nature. We demonstrate that Eri silkworm larvae could be used for production of VLPs of FMDV type O/IND/R2/75 for the first time. This approach could be useful for large scale production of recombinant VLPs for vaccine or diagnostic use in FMD control programme. PMID:26925448

  8. Properties of a recombinant bovine tissue factor expressed by Silkworm pupae and its performance as an Owren-type prothrombin time reagent for warfarin monitoring.

    PubMed

    Okuda, Masahiro; Taniguchi, Tomokuni; Takamiya, Osamu

    2012-09-01

    Tissue factor (TF), or thromboplastin, is a glycoprotein that triggers the extrinsic coagulation pathway. In blood coagulation testing, TF has been used as a natural source for determining Quick prothrombin time (PT) or the Owren PT (OBT). Currently, natural sources are being replaced with recombinant proteins because of their uniform characteristics and the possibility of stable mass production of PT reagents. Because bovine spongiform encephalopathy (BSE)-infected cows are widespread in Japan, we prepared a recombinant bovine TF (rbTF) with a baculovirus expression system using silkworms. To overcome the limitations of natural TF, especially in bovine brain, we expressed a full-length rbTF protein in Silkworm pupae with a baculovirus expression system. Baculovirus inactivation and the presence of DNA fragments in the rbTF fraction were confirmed using Reed-Muench and polymerase chain reaction methods after inactivation with a detergent. The rbTF fraction prepared by an immobilized anti-Silkworm pupae fluid protein Sepharose 4B column was identified as a visible band on western blots with a polyclonal antibody against human TF with cross-reactivity with TFs. The inhibition of the polyclonal antibody against human TF by the clotting assay for PT was identified, and amidolytic biological activity through activated factor VII on S-2288 substrate was observed. In conclusion, the rbTF expressed by the baculovirus system using Silkworm pupae was uniformly specific for bovine TF. The OBT reagent incorporated by this rbTF was similar to those of commercial reagents. It also showed a suitable International Sensitivity Index and reproducibility precision, thereby allowing for diagnostic use.

  9. [Cloning and expression profile of Bmyan in the silkworm (Bombyx mori) and experimental validation as one target of microRNA 7].

    PubMed

    Liu, Shiping; Huang, Yaxi; Yin, Jiyun; Wu, Xiaoyan; Zhou, Lanting; Wang, Wei; Xia, Qingyou

    2015-11-01

    microRNAs (miRNAs) are an extensive class of -22-nucleotide (nt) endogenous noncoding RNAs regulating life activities ofmetazoans through binding to 3'-untranslated regions (3'-UTRs) of their target genes. This work aimed to identify yan gene in the silkworm, reveal its expression profile and confirm if it is one target of bmo-miR-7 and, as such, have potential for contributing to better understanding of the molecular mechanisms involved in the metamorphosis of silkworm. Based on homolog searching and PCR amplification, we cloned the coding sequence (CDS) of Bmyan, which encodes 476 amino acid residues and contains SAM-PNT and ETs domains. Quantitative PCR (q-PCR), RT-PCR and microarray data revealed high expression of Bmyan in the head, body wall and ovary of day-3 fifth instar larval silkworm, low or no expression in other tissues. It was lowly expressed in the early larval stages, but highly expressed from late spinning to day 4 pupa. The 3'-UTR of Bmyan was obtained by rapid-amplification of cDNA ends (3'RACE) and predicted to contain two potential recognition sites of bmo-miR-7. The luciferase reporter vector containing the 3'-UTR of Bmyan was constructed and co-transfected into BmE cell line with the mimic of bmo-miR-7 and the decreased relative activity of luciferase showed that Bmyan is one target of bmo-miR-7. This work helps further functional analysis of bmo-miR-7 and Bmyan in the silkworm. PMID:26939444

  10. Effect of Induced Oxidative Stress and Herbal Extracts on Acid Phosphatase Activity in Lysosomal and Microsomal Fractions of Midgut Tissue of the Silkworm, Bombyx mori

    PubMed Central

    Gaikwad, Y. B.; Gaikwad, S. M.; Bhawane, G. P.

    2010-01-01

    Lysosomal and microsomal acid phosphatase activity was estimated in midgut tissue of silkworm larvae, Bombyx mori L. (Lepidoptera: Bombycidae), after induced oxidative stress by D-galactose. The larvae were simultaneously were treated with ethanolic extracts of Bacopa monniera and Lactuca sativa to study their antioxidant properties. Lipid peroxidation and fluorescence was measured to analyze extent of oxidative stress. The ethanolic extract of Lactuca sativa was found to be more effective in protecting membranes against oxidative stress than Bacopa monniera. PMID:20874583

  11. CRISPR/Cas9-mediated knockout of factors in non-homologous end joining pathway enhances gene targeting in silkworm cells.

    PubMed

    Zhu, Li; Mon, Hiroaki; Xu, Jian; Lee, Jae Man; Kusakabe, Takahiro

    2015-01-01

    Gene targeting can be achieved by precise genetic modifications through homology-directed repair (HDR) after DNA breaks introduced by genome editing tools such as CRISPR/Cas9 system. The most common form of HDR is homologous recombination (HR). Binding to the DNA breaks by HR factors is thought to compete with non-homologous end joining (NHEJ), an alternative DNA repair pathway. Here, we knocked out the factors in NHEJ by CRISPR/Cas9 system in silkworm cells, so that increased the activities of HR up to 7-fold. Also efficient HR-mediated genome editing events occurred between the chromosomal BmTUDOR-SN gene and donor DNA sequences with an EGFP gene in the middle of two homologous arms for the target gene. Utilizing the NHEJ-deficient silkworm cells, we found that homologous arms as short as 100 bp in donor DNA could be designed to perform precise genome editing. These studies should greatly accelerate investigations into genome editing of silkworm. PMID:26657947

  12. Evaluation of recombinant Neospora caninum antigens purified from silkworm larvae for the protection of N. caninum infection in mice.

    PubMed

    Yoshimoto, Mai; Otsuki, Takahiro; Itagaki, Kohei; Kato, Tatsuya; Kohsaka, Tetsuya; Matsumoto, Yumino; Ike, Kazunori; Park, Enoch Y

    2015-12-01

    Three antigens (NcSAG1, NcSRS2 and NcMIC3) from Neospora caninum were expressed using the BmNPV bacmid system in silkworm larvae and purified from the hemolymph. From 20 silkworm larvae, 1.5, 1.2 and 1.4 mg of purified recombinant NcSAG1, NcSRS2 and NcMIC3 were obtained, respectively. When each purified recombinant antigen was immunized with Freund's incomplete adjuvant (FIA) to mice, recombinant NcSAG1 induced a Th2 immune response in immunized mice and produced a SAG1-specific antibody. In the experiment where NcSAG1-immunized mice were challenged with N. caninum, the cerebral N. caninum burden was significantly reduced compared with that of either the FIA- or PBS-immunized mice. Recombinant NcSRS2 or NcMIC3 induced both Th1 and Th2 immune responses, but NcMIC3-immunization did not induce significant production of NcMIC3-specific antibodies. These results suggest that the silkworm can produce recombinant antigens of N. caninum, which can be used as a recombinant vaccine against N. caninum.

  13. Identification and analysis of the pigment composition and sources in the colored cocoon of the silkworm, Bombyx mori, by HPLC-DAD.

    PubMed

    Zhu, Lin; Zhang, Yu-Qing

    2014-01-01

    This study used the larval tissues and colored cocoons of silkworms, Bombyx mori L. (Lepidoptera: Bombycidae), that were fed leaves of cultivated mulberry, Husang 32, as experimental material. The pigment composition and content in colored cocoons and tissues of the 5th instar larvae and the mulberry leaves were rapidly detected using organic solvent extraction and reverse phase high-performance liquid chromatography with diode array detection. It was found that the mulberry leaf mainly contained four types of pigment: lutein (30.86%), β-carotene (26.3%), chlorophyll a (24.62%), and chlorophyll b (18.21%). The silk glands, blood, and cocoon shells of six yellow-red cocoons were used as the experimental materials. The results showed that there were generally two kinds of carotenoids (lutein and β-carotene) in the silk gland and cocoon shell, a little violaxanthin was detected in silk gland, and the pigment found in the blood was mainly lutein in all varieties of silkworm tested. Chlorophyll a and b had not been digested and utilized in the yellow-red series of silkworm. The method used to detect visible pigments reported here could be used to breed new colors of cocoons and to develop and utilize the pigments found in mulberry. PMID:25373178

  14. Production of Rous sarcoma virus-like particles displaying human transmembrane protein in silkworm larvae and its application to ligand-receptor binding assay.

    PubMed

    Tsuji, Yoshitaka; Deo, Vipin Kumar; Kato, Tatsuya; Park, Enoch Y

    2011-09-10

    Two types of Rous sarcoma virus (RSV) group-antigen protein (Gag) virus like particles (VLPs), full-length Gag (Gag701) and RSV protease domain (PR)-deleted mutant (Gag577) were expressed in silkworm larvae. Gag577 was secreted into hemolymph efficiently using wild type bacmid (WT), cysteine protease-deficient bacmid (CP(-)), cysteine protease and chitinase-deficient bacmid (CP(-)Chi(-)) bacmids, but comparatively Gag701 secretion levels were low. VLPs were purified on 10-60% (v/v) sucrose density gradient by ultracentrifugation and their structures confirmed under electron microscope. When hPRR and RSV Gag577 were co-expressed in silkworm larvae, human prorenin receptor (hPRR) was displayed on the surface of RSV VLPs, which was detected by Western blotting and immunoelectron microscopy. Moreover, binding of hPRR localized on the surface of VLPs to human prorenin was confirmed by ELISA. These results indicate that active hPRR was displayed on the surface of RSV VLPs, which can be utilized for drug discovery of hPRR blockers to prevent nephropathy. Moreover, this transmembrane protein display system using RSV Gag in silkworm larvae is applicable to expression of intact transmembrane proteins and binding assay of transmembrane proteins to its ligands, especially for transmembrane proteins which cannot be purified from membrane fractions in active states.

  15. Identification and analysis of the pigment composition and sources in the colored cocoon of the silkworm, Bombyx mori, by HPLC-DAD.

    PubMed

    Zhu, Lin; Zhang, Yu-Qing

    2014-01-01

    This study used the larval tissues and colored cocoons of silkworms, Bombyx mori L. (Lepidoptera: Bombycidae), that were fed leaves of cultivated mulberry, Husang 32, as experimental material. The pigment composition and content in colored cocoons and tissues of the 5th instar larvae and the mulberry leaves were rapidly detected using organic solvent extraction and reverse phase high-performance liquid chromatography with diode array detection. It was found that the mulberry leaf mainly contained four types of pigment: lutein (30.86%), β-carotene (26.3%), chlorophyll a (24.62%), and chlorophyll b (18.21%). The silk glands, blood, and cocoon shells of six yellow-red cocoons were used as the experimental materials. The results showed that there were generally two kinds of carotenoids (lutein and β-carotene) in the silk gland and cocoon shell, a little violaxanthin was detected in silk gland, and the pigment found in the blood was mainly lutein in all varieties of silkworm tested. Chlorophyll a and b had not been digested and utilized in the yellow-red series of silkworm. The method used to detect visible pigments reported here could be used to breed new colors of cocoons and to develop and utilize the pigments found in mulberry.

  16. Long-term preservation, regeneration, and cultivation of Paecilomyces tenuipes (Peck) Samson (Ascomycetes), an entomopathogenic fungus inoculated into the silkworm larva of Bombyx mori.

    PubMed

    Nam, Sung Hee; Li, Chun Ru; Li, Zeng-Zhi; Fan, Mei-Zhen; Kang, Seok Woo; Lee, Kwang Gill; Yeo, Joo Hong; Hwang, Jae Sam; Choi, Ji Young; Han, Sang Mi; Lee, Ki Man

    2011-01-01

    Paecilomyces tenuipes reportedly have anticancer and immune activities, along with various other medicinal uses. Cultured products with P. tenuipes are certified for use in food in South Korea, and processed goods containing this fungus have been developed in many countries, particularly South Korea, Japan, and China. Research on mass production technology-procured raw materials for the manufacture of P. tenuipes is very important; however, cultures of the fungus have been unstable. This study identified stable cultivation conditions, focusing on growth inhibition and revitalization. Moisture regulation and preservation of pupae inoculated with P. tenuipes were used to control growth inhibition and revitalization. When inoculated silkworm pupae were dehydrated to 4% moisture and preserved freeze-dried or at -70 degrees C, -20 degrees C, or 4 degrees C, the mycelia in their bodies were able to survive for 14 d. Inoculated silkworm pupae were rehydrated for 3 h and the mycelia within their bodies were recovered at 94.3-96.3%. Silkworm pupae at 4% moisture were able to survive for 135 d at temperatures < 4 degrees C and for 1 y after freeze-drying. Optimal conditions for synnemata induction were 25 degrees C and 100-300 1x.

  17. Mutation of a novel ABC transporter gene is responsible for the failure to incorporate uric acid in the epidermis of ok mutants of the silkworm, Bombyx mori.

    PubMed

    Wang, Lingyan; Kiuchi, Takashi; Fujii, Tsuguru; Daimon, Takaaki; Li, Muwang; Banno, Yutaka; Kikuta, Shingo; Kikawada, Takahiro; Katsuma, Susumu; Shimada, Toru

    2013-07-01

    ok mutants of the silkworm, Bombyx mori, exhibit highly translucent larval skin resulting from the inability to incorporate uric acid into the epidermal cells. Here we report the identification of a gene responsible for the ok mutation using positional cloning and RNAi experiments. In two independent ok mutant strains, we found a 49-bp deletion and a 233-bp duplication, respectively, in mRNAs of a novel gene, Bm-ok, which encodes a half-type ABC transporter, each of which results in translation of a truncated protein in each mutant. Although the Bm-ok sequence was homologous to well-known transporter genes, white, scarlet, and brown in Drosophila, the discovery of novel orthologs in the genomes of lepidopteran, hymenopteran, and hemipteran insects identifies it as a member of a new distinct subfamily of transporters. Embryonic RNAi of Bm-ok demonstrated that repression of Bm-ok causes a translucent phenotype in the first-instar silkworm larva. We discuss the possibility that Bm-ok forms a heterodimer with another half-type ABC transporter, Bmwh3, and acts as a uric acid transporter in the silkworm epidermis.

  18. Pigment pattern formation in the quail mutant of the silkworm, Bombyx mori: parallel increase of pteridine biosynthesis and pigmentation of melanin and ommochromes.

    PubMed

    Kato, Tomomi; Sawada, Hiroshi; Yamamoto, Takayuki; Mase, Keisuke; Nakagoshi, Motoko

    2006-08-01

    The larval pigment pattern in the silkworm, Bombyx mori, is formed by melanin, ommochromes and pteridines. During development all these pigments are synthesized autonomously, and possibly also with mutual interaction between them, to yield unique pigment patterns. In order to find the key trigger for such pigment pattern formation, developmental changes in pteridine biosynthesis were studied using the quail mutant (q/q), which has darker larval marks formed by melanin and an abundance of ommochromes in the integument. In the current study, emphasis has been placed on the analysis of GTP-cyclohydrolase I (GTP-CH I), which is a key enzyme for the biosynthesis of pteridines, during the development of the silkworm. Results of Northern blotting showed that in the quail mutant strong signals of GTP-CH I mRNA appeared around each period of ecdysis, while no such signals appeared in the background strain (+q/q) used. Also, both GTP-CH I activities and pteridine content were higher in the quail mutant compared with the background strain. These results strongly suggest that pteridine biosynthesis is closely linked to the formation of melanin and ommochromes. It is also suggested here that in the silkworm a recessive gene (q) may be involved in the regulation of its pigment pattern formation.

  19. Criticality emergency planning at Oak Ridge National Laboratory

    SciTech Connect

    Green, J.H.; Cain, G.C.

    1983-01-01

    A plan to protect personnel and control the spread of contamination in the event of a radiation accident at Oak Ridge National Laboratory (ORNL) is outlined. Procedures and personnel are presented. (ACK)

  20. 60 years of great science [Oak Ridge National Laboratory

    SciTech Connect

    2003-01-01

    This issue highlights Oak Ridge National Laboratory's contributions in more than 30 areas of research and related activities during the past 60 years and provides glimpses of current activities that are carrying on this heritage.

  1. Technical Evaluation of Oak Ridge Filter Test Facility

    SciTech Connect

    KRISKOVICH, J.R.

    2002-01-25

    Two evaluations of the Oak Ridge Department of Energy (DOE) Filter Test Facility (FTF) were performed on December 11 and 12, 2001, and consisted of a quality assurance and a technical evaluation. This report documents results of the technical evaluation.

  2. Science and Technology at Oak Ridge National Laboratory

    ScienceCinema

    Mason, Thomas

    2016-07-12

    ORNL Director Thom Mason explains the groundbreaking work in neutron sciences, supercomputing, clean energy, advanced materials, nuclear research, and global security taking place at the Department of Energy's Office of Science laboratory in Oak Ridge, Tenn.

  3. 97. HISTORIC PHOTOGRAPH, VIEW OF 'LAPHAM OAK,' NEW CANAAN, CA. ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    97. HISTORIC PHOTOGRAPH, VIEW OF 'LAPHAM OAK,' NEW CANAAN, CA. 1940. FROM PUBLIC WORKS COMMISSIONER'S REPORT. COLLECTION CONNECTICUT STATE LIBRARY AND ARCHIVES. - Merritt Parkway, Beginning in Greenwich & running 38 miles to Stratford, Greenwich, Fairfield County, CT

  4. Detail view of bronze door. Note oak branches with acorns ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Detail view of bronze door. Note oak branches with acorns in the left panels and olive branches with olives in right. - Flanders Field American Cemetery & Memorial, Chapel, Wortegemseweg 117, Waregem, West Flanders (Belgium)

  5. 19. DETAIL VIEW OF SKIFF BOW WITH OAK STEM AND ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    19. DETAIL VIEW OF SKIFF BOW WITH OAK STEM AND FRAMES PLANKED IN CEDAR USING COPPER CLINCH NAILS. TRANSOM OF SECOND SKIFF CAN BE SEEN BACKGROUND. - Lowell's Boat Shop, 459 Main Street, Amesbury, Essex County, MA

  6. Science and Technology at Oak Ridge National Laboratory

    SciTech Connect

    Mason, Thomas

    2012-11-01

    ORNL Director Thom Mason explains the groundbreaking work in neutron sciences, supercomputing, clean energy, advanced materials, nuclear research, and global security taking place at the Department of Energy's Office of Science laboratory in Oak Ridge, Tenn.

  7. The Oak Ridge Refrigerant Management Program

    NASA Technical Reports Server (NTRS)

    Kevil, Thomas H.

    1995-01-01

    For many years, chlorofluorocarbons (CFC's) have been used by the Department of Energy's (DOE) Oak Ridge Y-12 Plant in air conditioning and process refrigeration systems. However, Title 6 of the Clean Air Act Amendments (CAAA) and Executive Order 12843 (Procurement Requirements and Policies for Federal Agencies for Ozone Depleting Substances) signed by President Clinton require, as policy, that all federal agencies maximize their use of safe, alternate refrigerants and minimize, where economically practical, the use of Class 1 refrigerants. Unfortunately, many government facilities and industrial plants have no plan or strategy in place to make this changeover, even though their air conditioning and process refrigeration equipment may not be sustainable after CFC production ends December 31, 1995. The Y-12 Plant in Oak Ridge, Tennessee, has taken an aggressive approach to complying with the CAAA and is working with private industry and other government agencies to solve tough manufacturing and application problems associated with CFC and hydrochlorofluorocarbon (HCFC) alternatives. Y-12 was the first DOE Defense Program (DP) facility to develop a long-range Stratospheric Ozone Protection Plan for refrigerant management for compliance with the CAAA. It was also the first DOE DP facility to complete detailed engineering studies on retrofitting and replacing all air conditioning and process refrigeration equipment to enable operation with alternate refrigerants. The management plan and engineering studies are models for use by other government agencies, manufacturing plants, and private industry. This presentation identifies some of the hidden pitfalls to be encountered in the accelerated phaseout schedule of CFC's and explains how to overcome and prevent these problems. In addition, it outlines the general issues that must be considered when addressing the phase-out of ozone depleting substances and gives some 'lessons learned' by Y-12 from its Refrigerant Management

  8. The Oak Ridge Reservation Annual Site Environmental Report Summary, 2007

    SciTech Connect

    None, None

    2009-02-28

    The Oak Ridge Reservation Annual Site Environmental Report is prepared and published each year to inform the public of the environmental activities that take place on the reservation and in the surrounding areas. It is written to comply with DOE Order 231.1A, Environment, Safety, and Health Reporting. This document has been prepared to present the highlights of the Oak Ridge Reservation Annual Site Environmental Report 2007 in an easy-to-read, summary format.

  9. Decomposition of oak leaf litter and millipede faecal pellets in soil under temperate mixed oak forest

    NASA Astrophysics Data System (ADS)

    Tajovský, Karel; Šimek, Miloslav; Háněl, Ladislav; Šantrůčková, Hana; Frouz, Jan

    2015-04-01

    The millipedes Glomeris hexasticha (Diplopoda, Glomerida) were maintained under laboratory conditions and fed on oak leaf litter collected from a mixed oak forest (Abieto-Quercetum) in South Bohemia, Czech Republic. Every fourth day litter was changed and produced faecal pellets were separated and afterwards analysed. Content of organic carbon and C:N ratio lowered in faecal pellets as compared with consumed litter. Changes in content of chemical elements (P, K, Ca, Mg, Na) were recognised as those characteristic for the first stage of degradation of plant material. Samples of faecal pellets and oak leaf litter were then exposed in mesh bags between the F and H layers of forest soil for up to one year, subsequently harvested and analysed. A higher rate of decomposition of exposed litter than that of faecal pellets was found during the first two weeks. After 1-year exposure, the weight of litter was reduced to 51%, while that of pellets to 58% only, although the observed activity of present biotic components (algae, protozoans, nematodes; CO2 production, nitrogenase activity) in faecal pellets was higher as compared with litter. Different micro-morphological changes were observed in exposed litter and in pellets although these materials originated from the same initial sources. Comparing to intact leaf litter, another structural and functional processes occurred in pellets due to the fragmentation of plant material by millipedes. Both laboratory and field experiments showed that the millipede faecal pellets are not only a focal point of biodegradation activity in upper soil layers, but also confirmed that millipede feces undergo a slower decomposition than original leaf litter.

  10. Radioisotope production and management at Oak Ridge National Laboratory

    SciTech Connect

    Collins, E.D.; Aaron, W.S.; Alexander, C.W.; Bigelow, J.E.; Parks, J.T.; Tracy, J.G.; Wham, R.M.

    1994-09-01

    The production of radioisotopes has been one of the basic activities at Oak Ridge since the end of World War II. The importance of this work was best described by Alvin Weinberg, former Laboratory Director, when he wrote ``... If God has a golden book and writes down what it is that Oak Ridge National Laboratory did that had the biggest influence on science, I would guess that was the production and distribution of isotopes.`` Radioisotopes production continues to be an important aspect of Oak Ridge programs today and of those planned for the future. Past activities, current projects, and future plans and potentials will be described briefly in this paper. Also, some of the major issues facing the continued production of radioisotopes will be described. The scope of the program has always been primarily that of process development, followed by special batch-type productions, where no other supply exists. The technology developed has been available for adoption by US commercial corporations, and in cases where this has occurred, Oak Ridge has withdrawn as a supplier of the particular isotopes involved. One method of production that will not be described is that of target bombardment with an accelerator. This method was used at Oak Ridge prior to 1978 in the 86-inch Cyclotron. However, this method has not been used at Oak Ridge since then for radioisotope production, except as a research tool.

  11. Mesoscopic structural analysis of bedrock exposures at the Oak Ridge K-25 Site, Oak Ridge, Tennessee

    SciTech Connect

    Lemiszki, P.J.

    1995-07-01

    This document presents the detailed study of outcrop of mesoscopic structures during the geologic mapping completed in 1992-1993. The purpose of this study was to document the geometry and style of outcrop scale structures, such as fractures and faults and relate them to map scale structures present in the Oak Ridge K-25 Area. This report was prepared to document site characterization data collected during the scoping phase investigations in accordance with the requirements of the Comprehensive Environmental Response, Compensation, and Liability Act.

  12. Data Sharing Report Characterization of Isotope Row Facilities Oak Ridge National Laboratory Oak Ridge TN

    SciTech Connect

    Weaver, Phyllis C

    2013-12-12

    The U.S. Department of Energy (DOE) Oak Ridge Office of Environmental Management (EM-OR) requested that Oak Ridge Associated Universities (ORAU), working under the Oak Ridge Institute for Science and Education (ORISE) contract, provide technical and independent waste management planning support using funds provided by the American Recovery and Reinvestment Act (ARRA). Specifically, DOE EM-OR requested ORAU to plan and implement a survey approach, focused on characterizing the Isotope Row Facilities located at the Oak Ridge National Laboratory (ORNL) for future determination of an appropriate disposition pathway for building debris and systems, should the buildings be demolished. The characterization effort was designed to identify and quantify radiological and chemical contamination associated with building structures and process systems. The Isotope Row Facilities discussed in this report include Bldgs. 3030, 3031, 3032, 3033, 3033A, 3034, 3036, 3093, and 3118, and are located in the northeast quadrant of the main ORNL campus area, between Hillside and Central Avenues. Construction of the isotope production facilities was initiated in the late 1940s, with the exception of Bldgs. 3033A and 3118, which were enclosed in the early 1960s. The Isotope Row facilities were intended for the purpose of light industrial use for the processing, assemblage, and storage of radionuclides used for a variety of applications (ORNL 1952 and ORAU 2013). The Isotope Row Facilities provided laboratory and support services as part of the Isotopes Production and Distribution Program until 1989 when DOE mandated their shutdown (ORNL 1990). These facilities performed diverse research and developmental experiments in support of isotopes production. As a result of the many years of operations, various projects, and final cessation of operations, production was followed by inclusion into the surveillance and maintenance (S&M) project for eventual decontamination and decommissioning (D&D). The

  13. Geophysical Surveys of a Known Karst Feature, Oak Ridge Y-12 Plant, Oak Ridge, Tennessee

    SciTech Connect

    Doll, W.E.; Nyquist, J.E.; Carpenter, P.J.; Kaufmann, R.D.; Carr, B.J.

    1998-12-01

    Geophysical data were acquired at a site on the Oak Ridge Reservation, Tennessee to determine the characteristics of a mud-filled void and to evaluate the effectiveness of a suite of geophysical methods at the site. Methods that were used included microgravity, electrical resistivity, and seismic refraction. Both microgravity and resistivity were able to detect the void as well as overlying structural features. The seismic data provide bedrock depth control for the other two methods, and show other effects that are caused by the void.

  14. Project plan for the Background Soil Characterization Project on the Oak Ridge Reservation, Oak Ridge, Tennessee

    SciTech Connect

    Not Available

    1992-08-01

    The Background Soil characterization Project (BSCP) will provide background concentration levels of selected metals, organic compounds, and radionuclides in soils from uncontaminated on-site areas at the Oak Ridge Reservation (ORR), and off-site in the western part of Roane County and the eastern part of Anderson County. The BSCP will establish a database, recommend how to use the data for contaminated site assessment, and provide estimates of the potential human health and environmental risks associated with the background level concentrations of potentially hazardous constituents.

  15. INDEPENDENT VERIFICATION OF THE BUILDING 3550 SLAB AT OAK RIDGE NATIONAL LABORATORY OAK RIDGE, TENNESSEE

    SciTech Connect

    Weaver, Phyllis C.

    2012-05-08

    The Oak Ridge Institute for Science and Education (ORISE) has completed the independent verification survey of the Building 3550 Slab. The results of this effort are provided. The objective of this verification survey is to provide independent review and field assessment of remediation actions conducted by Safety and Ecology Corporation (SEC) to document that the final radiological condition of the slab meets the release guidelines. Verification survey activities on the Building 3550 Slab that included scans, measurements, and the collection of smears. Scans for alpha, alpha plus beta, and gamma activity identified several areas that were investigated.

  16. Geophysical Surveys of a Known Karst Feature, Oak Ridge Y-12 Plant, Oak Ridge, Tennessee

    SciTech Connect

    Carpenter, P.J.; Carr, B.J.; Doll, W.E.; Kaufmann, R.D.; Nyquist, J.E.

    1999-11-14

    Geophysical data were acquired at a site on the Oak Ridge Reservation, Tennessee to determine the characteristics of a mud-filled void and to evaluate the effectiveness of a suite of geophysical methods at the site. Methods that were used included microgravity, electrical resistivity, and seismic refraction. Both microgravity and resistivity were able to detect the void as well as overlying structural features. The seismic data provide bedrock depth control for the other two methods, and show other effects that are caused by the void.

  17. Leaf drop affects herbivory in oaks.

    PubMed

    Pearse, Ian S; Karban, Richard

    2013-11-01

    Leaf phenology is important to herbivores, but the timing and extent of leaf drop has not played an important role in our understanding of herbivore interactions with deciduous plants. Using phylogenetic general least squares regression, we compared the phenology of leaves of 55 oak species in a common garden with the abundance of leaf miners on those trees. Mine abundance was highest on trees with an intermediate leaf retention index, i.e. trees that lost most, but not all, of their leaves for 2-3 months. The leaves of more evergreen species were more heavily sclerotized, and sclerotized leaves accumulated fewer mines in the summer. Leaves of more deciduous species also accumulated fewer mines in the summer, and this was consistent with the idea that trees reduce overwintering herbivores by shedding leaves. Trees with a later leaf set and slower leaf maturation accumulated fewer herbivores. We propose that both leaf drop and early leaf phenology strongly affect herbivore abundance and select for differences in plant defense. Leaf drop may allow trees to dispose of their herbivores so that the herbivores must recolonize in spring, but trees with the longest leaf retention also have the greatest direct defenses against herbivores.

  18. Oak Ridge Reservation environmental report for 1990

    SciTech Connect

    Wilson, A.R.

    1991-09-01

    The purpose of this report is to provide information to the public about the impact of the US Department of Energy's (DOE's) facilities located on the Oak Ridge Reservation (ORR) on the public and the environment. It describes the environmental surveillance and monitoring activities conducted at and around the DOE facilities operated by Martin Marietta Energy Systems, Inc. Preparation and publication of this report is in accordance with DOE Order 5400.1. The order specifies a publication deadline of June of the following year for each calendar year of data. The primary objective of this report is to summarize all information collected for the previous calendar year regarding effluent monitoring, environmental surveillance, and estimates of radiation and chemical dose to the surrounding population. When multiple years of information are available for a program, trends are also evaluated. The first seven sections of Volume 1 of this report address this objective. The last three sections of Volume 1 provide information on solid waste management, special environmental studies, and quality assurance programs.

  19. Crush Testing at Oak Ridge National Laboratory

    SciTech Connect

    Feldman, Matthew R

    2011-01-01

    The dynamic crush test is required in the certification testing of some small Type B transportation packages. International Atomic Energy Agency regulations state that the test article must be 'subjected to a dynamic crush test by positioning the specimen on the target so as to suffer maximum damage.' Oak Ridge National Laboratory (ORNL) Transportation Technologies Group performs testing of Type B transportation packages, including the crush test, at the National Transportation Research Center in Knoxville, Tennessee (United States). This paper documents ORNL's experiences performing crush tests on several different Type B packages. ORNL has crush tested five different drum-type package designs, continuing its 60 year history of RAM package testing. A total of 26 crush tests have been performed in a wide variety of package orientations and crush plate CG alignments. In all cases, the deformation of the outer drum created by the crush test was significantly greater than the deformation damage caused by the 9 m drop test. The crush test is a highly effective means for testing structural soundness of smaller nondense Type B shipping package designs. Further regulatory guidance could alleviate the need to perform the crush test in a wide range of orientations and crush plate CG alignments.

  20. Leaf drop affects herbivory in oaks.

    PubMed

    Pearse, Ian S; Karban, Richard

    2013-11-01

    Leaf phenology is important to herbivores, but the timing and extent of leaf drop has not played an important role in our understanding of herbivore interactions with deciduous plants. Using phylogenetic general least squares regression, we compared the phenology of leaves of 55 oak species in a common garden with the abundance of leaf miners on those trees. Mine abundance was highest on trees with an intermediate leaf retention index, i.e. trees that lost most, but not all, of their leaves for 2-3 months. The leaves of more evergreen species were more heavily sclerotized, and sclerotized leaves accumulated fewer mines in the summer. Leaves of more deciduous species also accumulated fewer mines in the summer, and this was consistent with the idea that trees reduce overwintering herbivores by shedding leaves. Trees with a later leaf set and slower leaf maturation accumulated fewer herbivores. We propose that both leaf drop and early leaf phenology strongly affect herbivore abundance and select for differences in plant defense. Leaf drop may allow trees to dispose of their herbivores so that the herbivores must recolonize in spring, but trees with the longest leaf retention also have the greatest direct defenses against herbivores. PMID:23774946