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. PMID:21062145

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

    PubMed Central

    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

  3. Identification of Highly Active Genes from the Posterior Silk Gland of the Oak Silkworm, Antheraea yamamai, Through Expressed Sequence Tag (EST) Strategy

    Microsoft Academic Search

    Jae-Pil Jeon; Jae-Sam Hwang; Jeong-Hae Lee; Sang-Mong Lee; Hyun-Ah Kang; Uik Sohn

    2000-01-01

    Gene discovery from highly specialized tissues is important for understanding of development and tissue-dependent gene regulation in organism. To identify highly active genes from the posterior silk gland of the oak silkworm, Antheraea yamamai, which is specialized for the production of silk proteins, we constructed an oligo (dT) primed directional cDNA library from the posterior silk gland of the final

  4. Cloning of the fibroin gene from the oak silkworm, Antheraea yamamai and its complete sequence

    Microsoft Academic Search

    Jae-Sam Hwang; Jin-Sung Lee; Tae-Won Goo; Eun-Young Yun; Kwang-Sik Lee; Yong-Sung Kim; Byung-Rae Jin; Sang-Mong Lee; Keun-Young Kim; Seok-Woo Kang; Dong-Sang Suh

    2001-01-01

    The nucleotide sequences containing an entire genomic region and 5' upstream region of Antheraea yamamai fibroin gene have been determined. The gene consists of an initial exon encoding 14 amino acids, an intron (150 bp), and a long second exon coding for 2641 amino acids. The fibroin coding sequence shows a specialized organization with a highly repetitive region flanked by

  5. Molecular cloning and characterization of two 12 kDa FK506-binding protein genes in the Chinese oak silkworm, Antheraea pernyi.

    PubMed

    Chen, Mo; Chen, Miao-Miao; Yao, Rui; Li, Yan; Wang, Huan; Li, Yu-Ping; Liu, Yan-Qun

    2013-05-15

    Two 12 kDa FK506-binding protein (FKBP12) genes were isolated and characterized from Chinese oak silkworm Antheraea pernyi , an important agricultural and edible insect, designated ApFKBP12 A and B, respectively. Both ApFKBP12 A and B contained 108 amino acids with 82% sequence identity. Phylogenetic analysis showed that FKBP12 B sequences of A. pernyi, Bombyx mori , and Danaus plexippus were clearly separated from FKBP12 A sequences of these three species, suggesting that insect FKBP12 A and B may have been evolving independently. RT-PCR analyses revealed that two ApFKBP12 genes were expressed during the four developmental stages and in all tested tissues, and that the mRNA expression level of the ApFKBP12 A gene was significantly higher than that of the ApFKBP12 B gene. After heat shock treatment, expressions of the two FKBP12 genes were up-regulated, but at different time points. The results suggested that each paralogue of the FKBP12 genes may play a distinct functional role in the development of A. pernyi. PMID:23617895

  6. Diapause-Associated Protein3 Functions as Cu/Zn Superoxide Dismutase in the Chinese Oak Silkworm (Antheraea pernyi)

    PubMed Central

    Yu, Wei; Shu, Jianhong; Zhang, Yaozhou

    2014-01-01

    To better understand the molecular mechanism underlying of diapause in Antheraea pernyi (A.pernyi), we cloned a novel diapause-associated protein 3 (DAP3) gene from A.pernyi by reverse transcription-polymerase chain reaction (RT-PCR) and studied the biological functions. Sequence analysis revealed that this gene encodes 171 amino acids and has a conserved domain of Copper/Zinc superoxide dismutase (Cu/Zn-SOD). Western blot and qRT-PCR results showed that DAP3 was mainly expressed in the pupal stage, and gradually decreased as diapause development. DAP3 was also expressed in 1st and 5th instar larvae of A.pernyi. In tissues of 5th instar larvae of A.pernyi, DAP3 was mainly expressed in the epidermis, followed by the head, hemolymph and fat body. To identify the SOD activity of DAP3, we constructed a prokaryotic expression vector by inserting the coding region sequence into plasmid pET-28a (+) and obtained the purified rHIS-DAP3 fusion protein by Ni-NTA affinitive column. Importantly, we found the SOD activity of DAP3 fusion protein was approximately 0.6674 U/µg. To further confirm the SOD activity of DAP3 in vivo, we induced the oxidative stress model of pupae by UV irradiation. The results showed that both the mRNA and protein level of DAP3 significantly increased by UV irradiation. Furthermore, the SOD activity of the total lysate of pupae increased significantly at 10 min post UV irradiation and transiently returned to normal level afterwards. These results suggested that DAP3 might be a novel protein with SOD activity getting involved in regulation of diapause in A.pernyi. PMID:24613963

  7. 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. PMID:23706202

  8. Eicosanoids mediate sHSP 20.8 gene response to biotic stress in larvae of the Chinese oak silkworm Antheraea pernyi.

    PubMed

    Zhang, Cong-Fen; Dai, Li-Shang; Wang, Lei; Qian, Cen; Wei, Guo-Qing; Li, Jun; Zhu, Bao-Jian; Liu, Chao-Liang

    2015-05-10

    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 (A. pernyi; Lepidoptera: Saturniidae), cDNA encoding HSP20.8 in A. pernyi, termed Ap-sHSP20.8, was identified as a 564 bp ORF. The translated amino acid sequence encoded 187 residues with a calculated molecular mass of 20.8 kDa and an isoelectronic point (pI) of 5.98; the sequence showed homology to sHSP chaperone proteins from other insects. Ap-sHSP20.8 mRNA transcript expression was abundant in the midgut and fat body and found to be both constitutive and inducible by infectious stimuli. Therefore, Ap-sHSP20.8 may play important roles in A. pernyi immune responses under biotic stress. Furthermore, we found that eicosanoids could mediate the induction of Ap-sHSP20.8 in the fat body and midgut. Our findings show that sHSPs may be promising molecules to target in order to cripple immunity in insect pests. PMID:25527122

  9. 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. PMID:25280854

  10. Isolation, purification and characterization of silk protein sericin from cocoon peduncles of tropical tasar silkworm, Antheraea mylitta

    Microsoft Academic Search

    Rupesh Dash; Soumen Mukherjee; S. C. Kundu

    2006-01-01

    A high molecular weight water-soluble glue protein, sericin was identified in the cocoon peduncle (a strong thread connecting the cocoons to the branches of the tree with a ring) of the tropical tasar silkworm, Antheraea mylitta. The sericin was isolated by 8M urea containing 1% sodium dodecyl sulfate and ?-mercaptoethenol (2%) or by 1% sodium chloride. The protein was purified

  11. Biological relevance of host plant-derived terpenoid in the cocoons of the tropical tasar silkworm Antheraea mylitta

    Microsoft Academic Search

    P. C. Bindu; P. Jaisankar; F. Hauer; H. O. Gutzeit; S. C. Kundu

    2006-01-01

    We have characterized and studied the biological functions of a terpenoid derivative in the Indian tropical tasar silkworm, Antheraea mylitta reared on the primary host plant Arjun, Terminalia arjuna. The compound from insect cocoon turned out to be a terpenoid derivative which resembled oleanane type triterpene (Arjunolic acid) present in the host plant. The plant and cocoon compounds were anti-oxidative

  12. Identification of RAPD and SCAR markers associated with yield traits in the Indian tropical tasar silkworm Antheraea mylitta drury

    PubMed Central

    Dutta, Suhrid R.; Kar, Prasanta K.; Srivastava, Ashok K.; Sinha, Manoj K.; Shankar, Jai; Ghosh, Ananta K.

    2012-01-01

    The tropical tasar silkworm, Antheraea mylitta, is a semi-domesticated vanya silk-producing insect of high economic importance. To date, no molecular marker associated with cocoon and shell weights has been identified in this species. In this report, we identified a randomly amplified polymorphic DNA (RAPD) marker and examined its inheritance, and also developed a stable diagnostic sequence-characterized amplified region (SCAR) marker. Silkworms were divided into groups with high (HCSW) and low (LCSW) cocoon and shell weights, and the F2 progeny of a cross between these two groups were obtained. DNA from these silkworms was screened by PCR using 34 random primers and the resulting RAPD fragments were used for cluster analysis and discriminant function analysis (DFA). The clustering pattern in a UPGMA-based dendogram and DFA clearly distinguished the HCSW and LCSW groups. Multiple regression analysis identified five markers associated with cocoon and shell weights. The marker OPW16905 bp showed the most significant association with cocoon and shell weights, and its inheritance was confirmed in F2 progeny. Cloning and sequencing of this 905 bp fragment showed 88% identity between its 134 nucleotides and the Bmc-1/Yamato-like retroposon of A. mylitta. This marker was further converted into a diagnostic SCAR marker (SCOPW 16826 bp). The SCAR marker developed here may be useful in identifying the right parental stock of tasar silk-worms for high cocoon and shell weights in breeding programs designed to enhance the productivity of tasar silk. PMID:23271934

  13. Identification of RAPD and SCAR markers associated with yield traits in the Indian tropical tasar silkworm Antheraea mylitta drury.

    PubMed

    Dutta, Suhrid R; Kar, Prasanta K; Srivastava, Ashok K; Sinha, Manoj K; Shankar, Jai; Ghosh, Ananta K

    2012-12-01

    The tropical tasar silkworm, Antheraea mylitta, is a semi-domesticated vanya silk-producing insect of high economic importance. To date, no molecular marker associated with cocoon and shell weights has been identified in this species. In this report, we identified a randomly amplified polymorphic DNA (RAPD) marker and examined its inheritance, and also developed a stable diagnostic sequence-characterized amplified region (SCAR) marker. Silkworms were divided into groups with high (HCSW) and low (LCSW) cocoon and shell weights, and the F(2) progeny of a cross between these two groups were obtained. DNA from these silkworms was screened by PCR using 34 random primers and the resulting RAPD fragments were used for cluster analysis and discriminant function analysis (DFA). The clustering pattern in a UPGMA-based dendogram and DFA clearly distinguished the HCSW and LCSW groups. Multiple regression analysis identified five markers associated with cocoon and shell weights. The marker OPW16(905 bp) showed the most significant association with cocoon and shell weights, and its inheritance was confirmed in F(2) progeny. Cloning and sequencing of this 905 bp fragment showed 88% identity between its 134 nucleotides and the Bmc-1/Yamato-like retroposon of A. mylitta. This marker was further converted into a diagnostic SCAR marker (SCOPW 16(826 bp)). The SCAR marker developed here may be useful in identifying the right parental stock of tasar silk-worms for high cocoon and shell weights in breeding programs designed to enhance the productivity of tasar silk. PMID:23271934

  14. Comparative genetic diversity and genetic structure of three Chinese silkworm species Bombyx mori L. (Lepidoptera: Bombycidae), Antheraea pernyi Guérin-Meneville and Samia cynthia ricini Donovan (Lepidoptera: Saturniidae).

    PubMed

    Liu, Yan-Qun; Qin, Li; Li, Yu-Ping; Wang, Huan; Xia, Run-Xi; Qi, Yong-Hong; Li, Xi-Sheng; Lu, Cheng; Xiang, Zhong-Huai

    2010-01-01

    The genetic diversity and genetic structure of three Chinese silkworm species Bombyx mori L., Antheraea pernyi Guérin-Meneville and Samia cynthia ricini Donovan were comparatively assessed based on RAPD markers. At the species level, A. pernyi and B. mori showed high levels of genetic diversity, whereas S. cynthia ricini showed low level of genetic diversity. However, at the strain level, A. pernyi had relatively highest genetic diversity and B. mori had lowest genetic diversity. Analysis of molecular variance (AMOVA) suggested that 60% and 72% of genetic variation resided within strains in A. pernyi and S. cynthia ricini, respectively, whereas only 16% of genetic variation occurred within strains in B. mori. In UPGMA dendrogram, individuals of A. pernyi and B. mori formed the strain-specific genetic clades, whereas those of S. cynthia ricini were distributed in a mixed way. The implications of these results for the conservation and utilization in breeding programs of three silkworm species are discussed. PMID:21271066

  15. Silkworms

    NSDL National Science Digital Library

    Olivia Worland (Purdue University; Biological Sciences)

    2008-06-03

    Adult silkworm moths lay eggs to reproduce. The eggs hatch into silkworm larvae. The larvae spin silk cocoons and use them as they change from larvae to silkworm moths. Silkworm larvae exclusively eat mulberry leaves and their cocoons are used by human to make silk products such as silk fabric.

  16. Foliage of oaks grown under elevated CO2 reduces performance of Antheraea polyphemus (Lepidoptera: Saturniidae).

    PubMed

    Knepp, Rachel G; Hamilton, Jason G; Zangerl, Arthur R; Berenbaum, May R; DeLucia, Evan H

    2007-06-01

    To understand how the increase in atmospheric CO2 from human activity may affect leaf damage by forest insects, we examined host plant preference and larval performance of a generalist herbivore, Antheraea polyphemus Cram., that consumed foliage developed under ambient or elevated CO2. Larvae were fed leaves from Quercus alba L. and Quercus velutina Lam. grown under ambient or plus 200 microl/liter CO2 using free air carbon dioxide enrichment (FACE). Lower digestibility of foliage, greater protein precipitation capacity in frass, and lower nitrogen concentration of larvae indicate that growth under elevated CO2 reduced the food quality of oak leaves for caterpillars. Consuming leaves of either oak species grown under elevated CO2 slowed the rate of development of A. polyphemus larvae. When given a choice, A. polyphemus larvae preferred Q. velutina leaves grown under ambient CO2; feeding on foliage of this species grown under elevated CO2 led to reduced consumption, slower growth, and greater mortality. Larvae compensated for the lower digestibility of Q. alba leaves grown under elevated CO2 by increasing the efficiency of conversion of ingested food into larval mass. Despite equivalent consumption rates, larvae grew larger when they consumed Q. alba leaves grown under elevated compared with ambient CO2. Reduced consumption, slower growth rates, and increased mortality of insect larvae may explain lower total leaf damage observed previously in plots in this forest exposed to elevated CO2. By subtly altering aspects of leaf chemistry, the ever-increasing concentration of CO2 in the atmosphere will change the trophic dynamics in forest ecosystems. PMID:17540072

  17. The fibroin genes of Antheraea yamamai and Bombyx mori are different in their core regions but reveal a striking sequence similarity in their 5? ends and 5?flanking regions

    Microsoft Academic Search

    Toshiki Tamura; Hajime Inoue; Yoshiaki Suzuki

    1987-01-01

    We have cloned a fibroin gene from the Japanese oak silkworm, Antheraea yamamai, which belongs to the Saturniidae. The cloned DNA covers the entire region of the fibroin gene and a 1.4 kb 5'flanking sequence. Structural analysis of the gene by restriction mapping, S1 digestion and primer extension experiments shows that of Bombyx mori; both genes consist of one intron

  18. Purification and biochemical characterization of a 70 kDa sericin from tropical tasar silkworm, Antheraea mylitta

    Microsoft Academic Search

    Rupesh Dash; Sudip K. Ghosh; David L. Kaplan; S. C. Kundu

    2007-01-01

    Sericin isolated from the cocoon of the tropical tasar silkmoth Antheraea mylitta showed three major bands, with the lowest 70 kDa. This band was purified by anion exchange chromatography. Immunoblotting with concanavalin-A suggests a glycoprotein and CD analysis of secondary structure includes ?-sheet. Amino acid analysis shows that the protein is enriched in glycine and serine while the mole percentages of

  19. Comparative Study of Total Hydroperoxides and Antioxidant Defense System in the Indian Tropical Tasar Silkworm, Antheraea mylitta, in Diapausing and Non-Diapausing Generations

    PubMed Central

    Jena, Karmabeer; Kar, Prasanta K.; Babu, Chittithoti S.; Giri, Shantakar; Singh, Shyam S.; Prasad, Bhagwan C.

    2013-01-01

    In the present study, the total hydroperoxides, catalase, glutathione-s-transferase, and ascorbic acid contents were determined in different developmental stages of the non-diapause and the diapause generation of the tropical tasar silkworm, Antheraea mylitta Drury (Lepidoptera: Saturniidae). The results showed stage-specific significantly higher levels of total hydroperoxides, catalase, and ascorbic acid contents in the non-diapause as compared to the diapause generation (p < 0.05). However, a significantly enhanced level of glutathione-S-transferase activity was observed in mature 5th instar larvae of the diapause generation (p < 0.05). In the case of pupae, significantly higher levels of total hydroperoxides, catalase, and glutathione-s-transferase activity were observed in the non-diapause generation (p < 0.05). These results could be the effect of intensive metabolic transformation that takes place in tissues of the non-diapause generation and causes increased production of reactive oxygen species, such as hydroperoxides. The results suggest that antioxidants play an important role in protecting cells against reactive oxygen species. PMID:24786341

  20. Silkworm

    Microsoft Academic Search

    Yuji Yasukochi; Hiroshi Fujii; Marian R. Goldsmith

    The silkworm, Bombyx mori (n = 28), was domesticated from a mulberry-feeding moth, B. mandarina, in ancient China. Selective breeding, practiced from ancient times, and genetic studies performed since the early twentieth\\u000a century have yielded hundreds of practical strains with diverse economic characters, and linkage maps marked by > 200 loci\\u000a representing > 400 morphological and biochemical traits. Many types

  1. Foliage of Oaks Grown Under Elevated CO 2 Reduces Performance of Antheraea polyphemus (Lepidoptera: Saturniidae)

    Microsoft Academic Search

    Rachel G. Knepp; Jason G. Hamilton; Arthur R. Zangerl; May R. Berenbaum; Evan H. DeLucia

    2007-01-01

    To understand how the increase in atmospheric CO2 from human activity may affect leaf damage by forest insects, we examined host plant preference and larval performance of a generalist herbivore, Antheraea polyphemus Cram., that consumed foliage developed under ambient or elevated CO2. Larvae were fed leaves from Quercus alba L. and Quercus velutina Lam. grown under ambient or plus 200

  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. Bio-inspired fabrication of fibroin cryogels from the muga silkworm Antheraea assamensis for liver tissue engineering.

    PubMed

    Kundu, Banani; Kundu, S C

    2013-10-01

    Conventional scaffold fabrication techniques result in narrow pore architectures causing a limited interconnectivity and use of porogens, which affects the bio- or cyto-compatibility. To ameliorate this, cryogels are immensely explored due to their macro-porous nature, ease in fabrication, using ice crystals as porogens, the shape property, easy reproducibility and cost-effective fabrication technique. Cryogels in the present study are prepared from nonmulberry Indian muga silk gland protein fibroin of Antheraea assamensis using two different fabrication temperatures (-20 and -80 °C). Anionic surfactant sodium dodecyl sulfate is used to solubilize fibroin, which in turn facilitates gelation by accelerating the ß-sheet formation. Ethanol is employed to stabilize the 3D network and induces bimodal porosity. The gels thus formed demonstrate increased ß-sheet content (FTIR) and a considerable effect of pre-freezing temperatures on 3D micro-architectures. The cryogels are capable of absorbing large amounts of water and withstanding mechanical compression without structure deformation. Further, cell impregnated cryogels well support the viability of human hepatocarcinoma cells (live/dead assay). The formation of cellular aggregates (confocal laser and scanning electron microscope), derivation in metabolic activity and proliferation rate are obtained in constructs fabricated at different temperatures. In summary, the present work reveals promising insights in the development of a biomimetic functional template for biomedical therapeutics and liver tissue engineering. PMID:24002731

  4. Studies on the influence of host plants and effect of chemical stimulants on the feeding behavior in the muga silkworm, Antheraea assamensis.

    PubMed

    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 ?-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. Silkworm moths

    NSDL National Science Digital Library

    Gerd A.T. Müller (None; )

    2002-05-18

    Silkworm moths are the adult form of silkworm larvae. They emerge from the silk cocoons to mate. Mating is their only purpose and they do not eat or drink water. The females will lay hundreds of tiny white eggs.

  6. Silkworm cocoons

    NSDL National Science Digital Library

    Roman Neumüller (None; )

    2006-07-05

    Silkworm larvae spin silk cocoons to live in while they go through metamorphosis. They change from silkworm larvae into white silk moths. The silk cocoons are valuable to humans and can be made into silk fabric.

  7. Silk fibroin protein from mulberry and non-mulberry silkworms: cytotoxicity, biocompatibility and kinetics of L929 murine fibroblast adhesion

    Microsoft Academic Search

    Chitrangada Acharya; Sudip K. Ghosh; S. C. Kundu

    2008-01-01

    Silks fibers and films fabricated from fibroin protein of domesticated mulberry silkworm cocoon have been traditionally utilized\\u000a as sutures in surgery and recently as biomaterial films respectively. Here, we explore the possibility of application of silk\\u000a fibroin protein from non-mulberry silkworm cocoon as a potential biomaterial aid. In terms of direct inflammatory potential,\\u000a fibroin proteins from Antheraea mylitta and Bombyx

  8. Silkworm larvae

    NSDL National Science Digital Library

    Ma?gorzata Mi?aszewska (None; )

    2007-08-04

    Silkworm larvae hatch from eggs. They have 13 segments, split up into the head, thorax, and abdomen regions. The walking legs are on the thorax region and the prolegs are on the abdomen region. The larvae have a false eye on one of the segments to appear larger, spiracles on each segment to breathe through, and spinnerets to spin silk with near the head.

  9. PLANTINSECT INTERACTIONS Foliage of Oaks Grown Under Elevated CO2 Reduces Performance of

    E-print Network

    DeLucia, Evan H.

    ,weexaminedhostplantpreferenceandlarvalperformanceofageneralist herbivore, Antheraea polyphemus Cram., that consumed foliage developed under ambient or elevated CO2. LarvaePLANTÐINSECT INTERACTIONS Foliage of Oaks Grown Under Elevated CO2 Reduces Performance of Antheraea polyphemus (Lepidoptera: Saturniidae) RACHEL G. KNEPP,1 JASON G. HAMILTON,2 ARTHUR R. ZANGERL,3 MAY R

  10. Silkworm life cycle

    NSDL National Science Digital Library

    Hubert Ludwig (None; )

    2004-11-27

    Adult silkworms lay eggs to reproduce. Silkworm larvae hatch from these eggs. The larvae constantly eat only one thing- mulberry leaves. The larvae will spin silk cocoons for metamorphosis. The adults mate after emerging from the cocoon and the female will lay many small eggs.

  11. Transposable elements in commercially useful insects: I. Southern hybridization study of silkworms and honeybees using Drosophila probes.

    PubMed

    Kimura, K; Okumura, T; Ninaki, O; Kidwell, M G; Suzuki, K

    1993-02-01

    As a first step in surveying transposable elements in silkworms and honeybees, hybridization analyses were carried out using 16 known families of Drosophila transposable elements as probes. jockey and G were the only transposable elements that hybridized with genomic DNA of either honeybees or silkworms under the conditions of this study. jockey hybridized with genomic DNA of both European honeybees (Apis mellifera) and silkworms (Bombyx mori and Antheraea yamamai) and showed significant bands in Southern blots. Banding patterns were highly polymorphic. jockey did not, however, hybridize with any strains of the Asian honeybee (A. cerana). G elements showed a faint signal with the Asian honeybee, but not with any other insects tested. The results suggest that, even though it has some limitations, this approach can be used in practice as a first preliminary step in surveys for the presence of transposable elements in organisms which do not have good genetic information. PMID:8391286

  12. 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 ?-helix and ?-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. PMID:25532470

  13. Lipids of silkworm cocoons

    Microsoft Academic Search

    I. Tolibaev; Sh. R. Mad'yarov; A. I. Glushenkova

    1995-01-01

    The lipid complex of silkworm cocoons had been investigated. The qualitative and quantitative compositions of the total neutral lipids and glyco- and phospholipids have been determined. Considerable differences have been noted in the amounts of individual fatty acids in the neutral lipids and the phospholipids.

  14. Ligand binding to six recombinant pheromone-binding proteins of Antheraea polyphemus and Antheraea pernyi

    Microsoft Academic Search

    R. Maida; G. Ziegelberger; K.-E. Kaissling

    2003-01-01

    Binding properties of six heterologously expressed pheromone-binding proteins (PBPs) identified in the silkmoths Antheraea polyphemus and Antheraea pernyi were studied using tritium-labelled pheromone components, ( E, Z)-6,11-hexadecadienyl acetate ( 3H-Ac1) and ( E, Z)-6,11-hexadecadienal ( 3H-Ald), common to both species. In addition, a known ligand of PBP and inhibitor of pheromone receptor cells, the tritium-labelled esterase inhibitor decyl-thio-1,1,1-trifluoropropanone ( 3H-DTFP),

  15. Silk hydrogels from non-mulberry and mulberry silkworm cocoons processed with ionic liquids.

    PubMed

    Silva, Simone S; Popa, Elena G; Gomes, Manuela E; Oliveira, Mariana B; Nayak, Sunita; Subia, Bano; Mano, João F; Kundu, Subhas C; Reis, Rui L

    2013-11-01

    Matrices based on silk fibroin from the non-mulberry silkworm Antheraea mylitta and the mulberry silkworm Bombyx mori have demonstrated good applicability in regenerative medicine. However, the cocoons of A. mylitta are underutilized in part due to their lack of solubility in traditional organic solvents. Therefore, the present work investigates the solubilization and processing of degummed fibers obtained from the cocoons of both silkworm species into hydrogels using ionic liquids (ILs). The developed hydrogels exhibited a rubbery consistency, viscoelastic behavior and rapid degradation in the presence of protease XIV. Scanning electron and confocal microscopy images suggest that human adipose stem cells (hASCs) are able to adhere to and migrate at different levels within the hydrogel structures. Moreover, the MTS assay demonstrated the maintenance of cell metabolic activity for up to 28 days, while DNA quantification showed that hASCs were able to proliferate on the seeded hydrogels. The findings indicate that complete IL removal from the fabricated hydrogels results in a positive hASCs cellular response. Thus the present approach provides a unique opportunity to broaden the processability and application of silk fibroin obtained from A. mylitta cocoons for regenerative medicine, namely cartilage regeneration. PMID:23845228

  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 habitats. The information generated will be very useful in conservation of dwindling muga culture in Northeast India. PMID:22952746

  17. Extraction conditions of Antheraea mylitta sericin with high yields and minimum molecular weight degradation.

    PubMed

    Yun, Haesung; Oh, Hanjin; Kim, Moo Kon; Kwak, Hyo Won; Lee, Jeong Yun; Um, In Chul; Vootla, Shyam Kumar; Lee, Ki Hoon

    2013-01-01

    Although the technique for extracting the Bombyx mori sericin has been extensively known, the extraction of sericin from wild-silkworm cocoons is not yet standardized. The aim of this study was to find the optimal conditions for the extraction of sericin from Antheraea mylitta cocoons, with high yields and minimum degradation. We attempted to apply various protocols for the extraction of the A. mylitta sericin (AmS). Among these, we found that the extraction of AmS with a sodium carbonate solution exhibited the highest yield except the conventional soap-alkali extraction. To find the optimal conditions for the AmS extraction with the sodium carbonate, we changed the concentration of sodium carbonate and the treatment time. With an increase in the sodium carbonate concentration and the extraction time, the yield of AmS increased, but the molecular weight (MW) of AmS decreased. Considering the yield, molecular weight distribution (MWD) and amino acid composition of AmS, we suggest that the optimal conditions for the AmS extraction require treatment with 0.02 M sodium carbonate and boiling for 60 min. PMID:23026092

  18. KILLING SILKWORM COCOONS BY IRRADIATION

    Microsoft Academic Search

    Tsetskhladze

    1962-01-01

    The results of investigations of silkworm cocoon killing by irradiation ; are given. It is shown that the method is a very perspective one. Its ; industrial realization will give both increased raw silk yield and improved ; dynamometric characteristics of silk thread. (auth);

  19. Mechanical properties of silkworm cocoons

    Microsoft Academic Search

    Hong-Ping Zhao; Xi-Qiao Feng; Shou-Wen Yu; Wei-Zheng Cui; Feng-Zhu Zou

    2005-01-01

    Silkworm caterpillars, Bombyx mori, construct cocoons in order to protect their moth pupas against possible attacks from the outside. We experimentally measured the mechanical properties (Young's modulus, tensile strength, and thermo-mechanical parameters) of normal compact cocoons and the variations of these properties in the thickness direction of a cocoon. Tension tests were carried out by using rectangular specimens of two

  20. Comparative genomics of parasitic silkworm microsporidia reveal an association between genome expansion and host adaptation

    PubMed Central

    2013-01-01

    Background Microsporidian Nosema bombycis has received much attention because the pébrine disease of domesticated silkworms results in great economic losses in the silkworm industry. So far, no effective treatment could be found for pébrine. Compared to other known Nosema parasites, N. bombycis can unusually parasitize a broad range of hosts. To gain some insights into the underlying genetic mechanism of pathological ability and host range expansion in this parasite, a comparative genomic approach is conducted. The genome of two Nosema parasites, N. bombycis and N. antheraeae (an obligatory parasite to undomesticated silkworms Antheraea pernyi), were sequenced and compared with their distantly related species, N. ceranae (an obligatory parasite to honey bees). Results Our comparative genomics analysis show that the N. bombycis genome has greatly expanded due to the following three molecular mechanisms: 1) the proliferation of host-derived transposable elements, 2) the acquisition of many horizontally transferred genes from bacteria, and 3) the production of abundnant gene duplications. To our knowledge, duplicated genes derived not only from small-scale events (e.g., tandem duplications) but also from large-scale events (e.g., segmental duplications) have never been seen so abundant in any reported microsporidia genomes. Our relative dating analysis further indicated that these duplication events have arisen recently over very short evolutionary time. Furthermore, several duplicated genes involving in the cytotoxic metabolic pathway were found to undergo positive selection, suggestive of the role of duplicated genes on the adaptive evolution of pathogenic ability. Conclusions Genome expansion is rarely considered as the evolutionary outcome acting on those highly reduced and compact parasitic microsporidian genomes. This study, for the first time, demonstrates that the parasitic genomes can expand, instead of shrink, through several common molecular mechanisms such as gene duplication, horizontal gene transfer, and transposable element expansion. We also showed that the duplicated genes can serve as raw materials for evolutionary innovations possibly contributing to the increase of pathologenic ability. Based on our research, we propose that duplicated genes of N. bombycis should be treated as primary targets for treatment designs against pébrine. PMID:23496955

  1. Clicking caterpillars: acoustic aposematism in Antheraea polyphemus and other Bombycoidea

    Microsoft Academic Search

    Sarah G. Brown; George H. Boettner; Jayne E. Yack

    2007-01-01

    Acoustic signals produced by caterpillars have been documented for over 100 years, but in the majority of cases their significance is unknown. This study is the first to experimentally examine the phenomenon of audible sound production in larval Lepidoptera, focusing on a common silkmoth caterpillar, Antheraea polyphemus (Saturniidae). Larvae produce airborne sounds, resembling 'clicks', with their mandibles. Larvae typically signal

  2. Ligand binding to six recombinant pheromone-binding proteins of Antheraea polyphemus and Antheraea pernyi.

    PubMed

    Maida, R; Ziegelberger, G; Kaissling, K-E

    2003-09-01

    Binding properties of six heterologously expressed pheromone-binding proteins (PBPs) identified in the silkmoths Antheraea polyphemus and Antheraea pernyi were studied using tritium-labelled pheromone components, ( E, Z)-6,11-hexadecadienyl acetate ((3)H-Ac1) and ( E, Z)-6,11-hexadecadienal ((3)H-Ald), common to both species. In addition, a known ligand of PBP and inhibitor of pheromone receptor cells, the tritium-labelled esterase inhibitor decyl-thio-1,1,1-trifluoropropanone ((3)H-DTFP), was tested. The binding of ligands was measured after native gel electrophoresis and cutting gel slices. In both species, PBP1 and PBP3 showed binding of (3)H-Ac1. In competition experiments with (3)H-Ac1 and the third unlabelled pheromone component, ( E, Z)-4,9-tetradecadienyl acetate (Ac2), the PBP1 showed preferential binding of Ac1, whereas PBP3 preferentially bound Ac2. The PBP2 of both species bound (3)H-Ald only. All of the six PBPs strongly bound (3)H-DTFP. Among unlabelled pheromone derivatives, alcohols were revealed to be the best competitors for (3)H-Ac1 and (3)H-Ald bound to PBPs. No pH influence was found for (3)H-Ac1 binding to, or its release from, the PBP3 of A. polyphemus and A. pernyi between pH 4.0 and pH 7.5. The data indicate binding preference of each of the three PBP-subtypes (1-3) for a specific pheromone component and support the idea that PBPs contribute to odour discrimination, although to a smaller extent than receptor activation. PMID:12879348

  3. TREATMENT OF SILKWORM COCOONS BY RADIATION

    Microsoft Academic Search

    U. A. Ariffov; I. D. Artmeladze; V. A.. Barnov

    1959-01-01

    The existing methods of silkworm cocoon treatment, including heat, ; chemicals, and vacuum, essentially damage the silk thread and considerably ; decrease the yield of raw silk as compared with untreated cocoons. A possibility ; of treating the silkworm chrysalis by Co⁶° gamma rays was studied in this ; connection. A comparative study of the properties of the silk thread

  4. ASPERGILLUS BOMBYCIS GENOTYPES (RFLP) FROM SILKWORM CULTIVATION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eighteen isolates of Aspergillus bombycis from samples of dust, insect frass, and soil collected from 8 silkworm rearing facilities in Japan, as well as single silkworm rearing facilities in Indonesia and Malaysia, were subjected to DNA fingerprinting. PstI digests of total genomic DNA from each is...

  5. Phylogeny and evolutionary history of the silkworm.

    PubMed

    Sun, Wei; Yu, Hongsong; Shen, Yihong; Banno, Yutaka; Xiang, Zhonghuai; Zhang, Ze

    2012-06-01

    The silkworm, Bombyx mori, played an important role in the old Silk Road that connected ancient Asia and Europe. However, to date, there have been few studies of the origins and domestication of this species using molecular methods. In this study, DNA sequences of mitochondrial and nuclear loci were used to infer the phylogeny and evolutionary history of the domesticated silkworm and its relatives. All of the phylogenetic analyses indicated a close relationship between the domesticated silkworm and the Chinese wild silkworm. Domestication was estimated to have occurred about 4100 years ago (ya), and the radiation of the different geographic strains of B. mori about 2000 ya. The Chinese wild silkworm and the Japanese wild silkworm split about 23600 ya. These estimates are in good agreement with the fossil evidence and historical records. In addition, we show that the domesticated silkworm experienced a population expansion around 1000 ya. The divergence times and the population dynamics of silkworms presented in this study will be useful for studies of lepidopteran phylogenetics, in the genetic analysis of domestic animals, and for understanding the spread of human civilizations. PMID:22744178

  6. 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

  7. Segmental duplications in the silkworm genome

    PubMed Central

    2013-01-01

    Background Segmental duplications (SDs) or low-copy repeats play important roles in both gene and genome evolution. SDs have been extensively investigated in many organisms, however, there is no information about SDs in the silkworm, Bombyx mori. Result In this study, we identified and annotated the SDs in the silkworm genome. Our results suggested that SDs constitute ~1.4% of the silkworm genome sequence (?1 kb in length and ?90% in the identity of sequence); the number is similar to that in Drosophila melanogaster but smaller than mammalian organisms. Almost half (42%) of the SD sequences are not assigned to chromosomes, indicating that the SDs are challenges for the assembling of genome sequences. We also provided experimental validation of large duplications using qPCR. The analysis of SD content indicated that the genes related to immunity, detoxification, reproduction, and environmental signal recognition are significantly enriched in the silkworm SDs. Conclusion Our results suggested that segmental duplications have been problematic for sequencing and assembling of the silkworm genome. SDs may have important biological significances in immunity, detoxification, reproduction, and environmental signal recognition in the silkworm. This study provides insight into the evolution of the silkworm genome and an invaluable resource for insect genomics research. PMID:23901934

  8. Molecular characterization of genome segments 1 and 3 encoding two capsid proteins of Antheraea mylitta cytoplasmic polyhedrosis virus

    PubMed Central

    2010-01-01

    Background Antheraea mylitta cytoplasmic polyhedrosis virus (AmCPV), a cypovirus of Reoviridae family, infects Indian non-mulberry silkworm, Antheraea mylitta, and contains 11 segmented double stranded RNA (S1-S11) in its genome. Some of its genome segments (S2 and S6-S11) have been previously characterized but genome segments encoding viral capsid have not been characterized. Results In this study genome segments 1 (S1) and 3 (S3) of AmCPV were converted to cDNA, cloned and sequenced. S1 consisted of 3852 nucleotides, with one long ORF of 3735 nucleotides and could encode a protein of 1245 amino acids with molecular mass of ~141 kDa. Similarly, S3 consisted of 3784 nucleotides having a long ORF of 3630 nucleotides and could encode a protein of 1210 amino acids with molecular mass of ~137 kDa. BLAST analysis showed 20-22% homology of S1 and S3 sequence with spike and capsid proteins, respectively, of other closely related cypoviruses like Bombyx mori CPV (BmCPV), Lymantria dispar CPV (LdCPV), and Dendrolimus punctatus CPV (DpCPV). The ORFs of S1 and S3 were expressed as 141 kDa and 137 kDa insoluble His-tagged fusion proteins, respectively, in Escherichia coli M15 cells via pQE-30 vector, purified through Ni-NTA chromatography and polyclonal antibodies were raised. Immunoblot analysis of purified polyhedra, virion particles and virus infected mid-gut cells with the raised anti-p137 and anti-p141 antibodies showed specific immunoreactive bands and suggest that S1 and S3 may code for viral structural proteins. Expression of S1 and S3 ORFs in insect cells via baculovirus recombinants showed to produce viral like particles (VLPs) by transmission electron microscopy. Immunogold staining showed that S3 encoded proteins self assembled to form viral outer capsid and VLPs maintained their stability at different pH in presence of S1 encoded protein. Conclusion Our results of cloning, sequencing and functional analysis of AmCPV S1 and S3 indicate that S3 encoded viral structural proteins can self assemble to form viral outer capsid and S1 encoded protein remains associated with it as inner capsid to maintain the stability. Further studies will help to understand the molecular mechanism of capsid formation during cypovirus replication. PMID:20684765

  9. 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...

  10. 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

  11. 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. PMID:26044642

  12. Mechanical properties of silkworm cocoon pelades

    Microsoft Academic Search

    Hong-Ping Zhao; Xi-Qiao Feng; Wei-Zheng Cui; Feng-Zhu Zou

    2007-01-01

    The pelade, the innermost layer of silkworm cocoon next to the chrysalis, has special microstructures, mechanical properties and protective functions distinctly different from those of all the other layers. In the present paper, a series of static tensile tests and dynamic mechanical thermal analysis were performed for the first time to measure the mechanical properties of pelades, including Young’s modulus,

  13. Snmp-1, a Novel Membrane Protein of Olfactory Neurons of the Silk Moth Antheraea polyphemus with Homology to the CD36 Family of

    E-print Network

    Vogt, Richard G.

    Snmp-1, a Novel Membrane Protein of Olfactory Neurons of the Silk Moth Antheraea polyphemus of the wild silk moth Antheraea polyphemus. We have purified and cloned a prominent 67-kDa protein which we of olfactory neuron receptor membranes of the wild silk moth Antheraea polyphemus. The morphology of the A

  14. Screening Strains of the Mulberry Silkworm, Bombyx mori, for Thermotolerance

    PubMed Central

    Kumari, Savarapu Sugnana; Subbarao, Sure Venkata; Misra, Sunil; Murty, Upadyayula Suryanarayana

    2011-01-01

    A tropical climate prevails in most of the sericultural areas in India, where temperature increases during the summer lead to adverse effects on temperate bivoltine silkworm rearing and cause crop losses. Screening for thermotolerance in the silkworm, Bombyxmori L. (Lepidoptera: Bombycidae) is an essential prerequisite for the development of thermotolerant breeds/hybrids. In the current study, the aim was to identify potential bivoltine silkworm strains specific for tolerance to high temperature. The third day of fifth stage silkworm larvae of bivoltine strains were subjected to high temperature of 36 ± 1° C with RH of 50 ± 5 % for six hours (10:00–16:00) every day until spinning for three consecutive generations. Highly significant differences were found among all genetic traits of bivoltine silkworm strains in the treated groups. Three groups of silkworm resulted including susceptible, moderately tolerant, and tolerant by utilizing pupation rate or survival rate with thermal stress as the index for thermotolerance. Furthermore, based on the overall silkworm rearing performance of nine quantitative genetic traits such as larval weight, cocoon yield by number and weight, pupation, single cocoon and shell weight, shell ratio, filament length and denier, three bivoltine silkworm strains, BD2-S, SOF-BR and BO2 were developed as having the potential for thermotolerance. The data from the present study enhance knowledge for the development of thermo tolerant silkworm breeds/ hybrids and their effective commercial utilization in the sericulture industry. PMID:22225406

  15. Reconstruction and morphometry of silkmoth olfactory hairs: A comparative study of sensilla trichodea on the antennae of male Antheraea polyphemus and Antheraea pernyi (Insecta, Lepidoptera)

    Microsoft Academic Search

    T. A. Keil

    1984-01-01

    Olfactory trichoid hairs on the antennae of male Antheraea silkmoths were reconstructed with respect to the following parameters: number, shape, course, and dimensions of outer dendritic segments as well as the numbers of their microtubules; inner and outer dimensions of the cuticular hair shafts; and number and distribution of pores and pore tubules in the hair walls. The smallest distances

  16. Threshold and odor specificity of pheromone-sensitive neurons in the deutocerebrum of Antheraea pernyi and A. polyphemus (Saturnidae)

    Microsoft Academic Search

    J. Boeckh; V. Boeckh

    1979-01-01

    In extracellular recordings neurons were located in the deutocerebrum ofAntheraea pernyi andAntheraea polyphemus which are specifically excited by components found in the female sexual attractant glands of both species (Figs. 3, 4). Extracellular cobalt injections via the recording electrode revealed a certain morphological type of neuron with widely branching arborizations in a big mass of neuropile (“macroglomerulus”) in the dorsal

  17. ?-Fructofuranosidase Genes of the Silkworm, Bombyx mori

    PubMed Central

    Daimon, Takaaki; Taguchi, Tomohiro; Meng, Yan; Katsuma, Susumu; Mita, Kazuei; Shimada, Toru

    2008-01-01

    Mulberry latex contains extremely high concentrations of alkaloidal sugar mimic glycosidase inhibitors, such as 1,4-dideoxy-1,4-imino-d-arabinitol (d-AB1) and 1-deoxynojirimycin (DNJ). Although these compounds do not harm the silkworm, Bombyx mori, a mulberry specialist, they are highly toxic to insects that do not normally feed on mulberry leaves. d-AB1 and DNJ are strong inhibitors of ?-glucosidases (EC 3.2.1.20); however, they do not affect the activity of?-fructofuranosidases (EC 3.2.1.26). Although?-glucosidase genes are found in a wide range of organisms, ?-fructofuranosidase genes have not been identified in any animals so far. In this study, we report the identification and characterization of ?-fructofuranosidase genes (BmSuc1 and BmSuc2) from B. mori. The BmSuc1 gene was highly expressed in the midgut and silk gland, whereas the expression of BmSuc2 gene was not detected. BmSuc1 encodes a functional ?-fructofuranosidase, whose enzymatic activity was not inhibited by DNJ or d-AB1. We also showed that BmSUC1 protein localized within the midgut goblet cell cavities. Collectively, our data clearly demonstrated that BmSuc1 serves as a sugar-digesting enzyme in the silkworm physiology. This anomalous presence of the ?-fructofuranosidase gene in the B. mori genome may partly explain why the silkworm can circumvent the mulberry's defense system. PMID:18397891

  18. Analysis on Gender of Silkworms by MRI Technology

    Microsoft Academic Search

    Cong Liu; Zhao Hui Ren; Hong Zhi Wang; Pei Qiang Yang; Xue Long Zhang

    2008-01-01

    At present silkworm cocoons, dried, boiled and reeled, are the female and male mix, so it's very difficult to produce high grade raw silk in large quantity. In the progress of the silk reeling what we can do to separate the complex of female and male silkworms still encounters presently certain difficulties. In this paper the intrinsic relaxation time of

  19. Overexpression of host plant urease in transgenic silkworms.

    PubMed

    Jiang, Liang; Huang, Chunlin; Sun, Qiang; Guo, Huizhen; Peng, Zhengwen; Dang, Yinghui; Liu, Weiqiang; Xing, Dongxu; Xu, Guowen; Zhao, Ping; Xia, Qingyou

    2015-06-01

    Bombyx mori and mulberry constitute a model of insect-host plant interactions. Urease hydrolyzes urea to ammonia and is important for the nitrogen metabolism of silkworms because ammonia is assimilated into silk protein. Silkworms do not synthesize urease and acquire it from mulberry leaves. We synthesized the artificial DNA sequence ure-as using the codon bias of B. mori to encode the signal peptide and mulberry urease protein. A transgenic vector that overexpresses ure-as under control of the silkworm midgut-specific P2 promoter was constructed. Transgenic silkworms were created via embryo microinjection. RT-PCR results showed that urease was expressed during the larval stage and qPCR revealed the expression only in the midgut of transgenic lines. Urea concentration in the midgut and hemolymph of transgenic silkworms was significantly lower than in a nontransgenic line when silkworms were fed an artificial diet. Analysis of the daily body weight and food conversion efficiency of the fourth and fifth instar larvae and economic characteristics indicated no differences between transgenic silkworms and the nontransgenic line. These results suggested that overexpression of host plant urease promoted nitrogen metabolism in silkworms. PMID:25549597

  20. Clicking caterpillars: acoustic aposematism in Antheraea polyphemus and other Bombycoidea.

    PubMed

    Brown, Sarah G; Boettner, George H; Yack, Jayne E

    2007-03-01

    Acoustic signals produced by caterpillars have been documented for over 100 years, but in the majority of cases their significance is unknown. This study is the first to experimentally examine the phenomenon of audible sound production in larval Lepidoptera, focusing on a common silkmoth caterpillar, Antheraea polyphemus (Saturniidae). Larvae produce airborne sounds, resembling ;clicks', with their mandibles. Larvae typically signal multiple times in quick succession, producing trains that last over 1 min and include 50-55 clicks. Individual clicks within a train are on average 24.7 ms in duration, often consisting of multiple components. Clicks are audible in a quiet room, measuring 58.1-78.8 dB peSPL at 10 cm. They exhibit a broadband frequency that extends into the ultrasound spectrum, with most energy between 8 and 18 kHz. Our hypothesis that clicks function as acoustic aposematic signals, was supported by several lines of evidence. Experiments with forceps and domestic chicks correlated sound production with attack, and an increase in attack rate was positively correlated with the number of signals produced. In addition, sound production typically preceded or accompanied defensive regurgitation. Bioassays with invertebrates (ants) and vertebrates (mice) revealed that the regurgitant is deterrent to would-be predators. Comparative evidence revealed that other Bombycoidea species, including Actias luna (Saturniidae) and Manduca sexta (Sphingidae), also produce airborne sounds upon attack, and that these sounds precede regurgitation. The prevalence and adaptive significance of warning sounds in caterpillars is discussed. PMID:17337712

  1. Synnemata Production Using Silkworm Variety, Female Yangwonjam by Isaria tenuipes.

    PubMed

    Ji, Sang-Duk; Sung, Gyoo-Byung; Kang, Pil-Don; Kim, Kee-Young; Choi, Yong-Soo; Kim, Nam-Suk; Woo, Soon-Ok; Han, Sang-Mi; Hong, In-Pyo; Ha, Nam-Gyu

    2011-09-01

    This study was conducted to confirm the possible use of female Yangwonjam as a host for synnemata production of Isaria tenuipes in eight local areas in Korea. Silkworm pupation rate, infection rate and synnemata characteristics of I. tenuipes were examined. Normal silkworms had a higher pupation rate than silkworms inoculated with I. tenuipes. The pupae survival percentage of normal silkworm in cocoons was 92.5~97.6%, whereas it ranged from 91.1~95.6% in silkworms sprayed with I. tenuipes. Female Yangwonjam showed the highest survival percentage at 97.6% among the silkworm varieties tested. I. tenuipes infection rate of larvae of 5th instar newly-exuviated silkworms was 89.2~90.7% in the spring rearing season and 98.2~99.3% in the autumn rearing season. Synnemata production of I. tenuipes was excellent in female Yangwonjam with an incidence rate of 98.0% followed by male Yangwonjam (94.1%) and Baegokjam (93.3%) in the spring rearing season. Synnemata living weight ranged from 1.44~0.94 g in the spring rearing season. The female Yangwonjam had the heaviest synnemata weight (1.44 g) in the spring rearing season. The synnemata of I. tenuipes produced on pupae were white or milky-white in color, and were similar in shape and color to wild synnemata collected in Korea. PMID:22783097

  2. Synnemata Production Using Silkworm Variety, Female Yangwonjam by Isaria tenuipes

    PubMed Central

    Ji, Sang-Duk; Sung, Gyoo-Byung; Kang, Pil-Don; Kim, Kee-Young; Choi, Yong-Soo; Kim, Nam-Suk; Woo, Soon-Ok; Han, Sang-Mi; Ha, Nam-Gyu

    2011-01-01

    This study was conducted to confirm the possible use of female Yangwonjam as a host for synnemata production of Isaria tenuipes in eight local areas in Korea. Silkworm pupation rate, infection rate and synnemata characteristics of I. tenuipes were examined. Normal silkworms had a higher pupation rate than silkworms inoculated with I. tenuipes. The pupae survival percentage of normal silkworm in cocoons was 92.5~97.6%, whereas it ranged from 91.1~95.6% in silkworms sprayed with I. tenuipes. Female Yangwonjam showed the highest survival percentage at 97.6% among the silkworm varieties tested. I. tenuipes infection rate of larvae of 5th instar newly-exuviated silkworms was 89.2~90.7% in the spring rearing season and 98.2~99.3% in the autumn rearing season. Synnemata production of I. tenuipes was excellent in female Yangwonjam with an incidence rate of 98.0% followed by male Yangwonjam (94.1%) and Baegokjam (93.3%) in the spring rearing season. Synnemata living weight ranged from 1.44~0.94 g in the spring rearing season. The female Yangwonjam had the heaviest synnemata weight (1.44 g) in the spring rearing season. The synnemata of I. tenuipes produced on pupae were white or milky-white in color, and were similar in shape and color to wild synnemata collected in Korea. PMID:22783097

  3. 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 ?-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

  4. 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 ?-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

  5. Elektronenmikroskopische Beobachtungen über die Fusion myogener Zellen bei Antheraea polyphemus (Lepidoptera)

    Microsoft Academic Search

    R. F. Stocker

    1974-01-01

    Summary The fusion of both myoblasts and ‘Muskelanlagezellen’ during development of the dorsolongitudinal muscles inAntheraea polyphemus (Lep.) is shown at the ultrastructural level. Several cytoplasmic bridges are formed simultaneously between the two cells. It is supposed that the effect of breakdown of both myoblast membrane and sarcolemma originates from the myoblast.

  6. Similarity of reaction spectra and odor discrimination: single receptor cell recordings in Antheraea polyphemus (Saturniidae)

    Microsoft Academic Search

    Wolf A. Kafka

    1987-01-01

    In the general context of primary olfactory coding, the reaction spectra of 50 receptor cells to a total of 54 substances were electrophysiologically determined on base of dose response curves and quantitatively compared with regard to their similarity by methods of vector analysis. The receptor cells are distributed among 3 morphologically distinguishable sensilla on the antennae of the male silkmothAntheraea

  7. HORMONAL CONTROL OF MUSCLE ATROPHY AND DEGENERATION IN THE MOTH ANTHERAEA POLYPHEMUS

    Microsoft Academic Search

    LAWRENCE M. SCHWARTZ; JAMES W. TRUMAN

    1984-01-01

    SUMMARY 1. The intersegmental muscles of the giant silkmoth Antheraea polyphemus (Cramer) can undergo two forms of degenerative changes: a wasting atrophy that lasts about 6 days or rapid dissolution that is completed within 30 h. 2. Muscle atrophy is induced by a dramatic decline in the endogenous titres of the steroid moulting hormone 20-hydroxyecdysone. 3. 20-Hydroxyecdysone appears to act

  8. Functional morphology of the ommatidia in the compound eye of the moth, Antheraea polyphemus (Insecta, Saturniidae)

    Microsoft Academic Search

    Friederike Anton-Erxleben; Helmut Langer

    1988-01-01

    The fine structure of the superposition eye of the Saturniid moth Antheraea polyphemus Cramer was investigated by electron microscopy. Each of the approximately 10000 ommatidia consists of the same structural components, but regarding the arrangement of the ommatidia and the rhabdom structure therein, two regions of the eye have to be distinguished. In a small dorsal rim area, the ommatidia

  9. Preparation and physical properties of antheraea yamamai\\/Bombyx mori silk fibroin blending film

    Microsoft Academic Search

    Linlin Lv; Yanqiong Wei; Jiannan Wang; Mingzhong Li; Huijing Zhao; Guiyang Liu; Qiang Lv

    2011-01-01

    Antheraea yamamai (A yamamai) silk fibroin contains tripeptide sequence of Arg-Gly-Asp (RGD) and more positively charged amino acids residues. In this study, aqueous A. yamamai silk fibroin solution and Bombyx mori (B. mori) silk fibroin solution in various blending ratios were mixed to prepare blending films. The morphology and physical prosperities of the blending films were examined and analyzed through

  10. Cyclic GMP Levels and Guanylate Cyclase Activity in Pheromone Sensitive Antennae of the Silkmoths Antheraea polyphemus and Bombyx mori

    Microsoft Academic Search

    G. Ziegelberger; M. J. van den Berg; K.-E. Kaissling; S. Klumpp; J. E. Schultz

    Female sex pheromones applied to freshly isolated, living antennae of male Antheraea polyphemus and Bombyx mori led to an increase of cGMP. A 1:l mixture of 2 pheromone components of Antheraea polyphemus blown for 10 set in physiological concentrations over their antenna1 branches raised cGMP levels about 1.34-fold (kO.08 SEM, n = 23) from a basal level of 3.0 f

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

    Microsoft Academic Search

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

    2010-01-01

    Many recombinant proteins have been successfully produced in silkworm larvae or pupae and used for academic and industrial\\u000a purposes. Several recombinant proteins produced by silkworms have already been commercialized. However, construction of a\\u000a recombinant baculovirus containing a gene of interest requires tedious and troublesome steps and takes a long time (3–6 months).\\u000a The recent development of a bacmid, Escherichia coli and

  12. Transgenic silkworms that weave recombinant proteins into silk cocoons

    Microsoft Academic Search

    Masahiro Tomita

    2011-01-01

    As a result of breeding for more than 4,000 years, the silkworm, Bombyx mori, has acquired the ability to synthesize bulk amounts of silk proteins in its silk glands. To utilize this capacity for mass\\u000a production of useful proteins, transgenic silkworms were generated that synthesized recombinant proteins in the silk gland\\u000a and secreted them into the silk cocoon. The silk gland

  13. Mechanism of fluorescent cocoon sex identification for silkworms Bombyx mori

    Microsoft Academic Search

    YuQing Zhang; XiaoHua Yu; WeiDe Shen; YongLei Ma; LiXia Zhou; NaiXi Xu; ShuQian Yi

    2010-01-01

    By using silkworms, Bombyx mori, fluorescent cocoon sex identification (FCSI) as an experimental material, direct fluorescence spectrometry of the cocoon\\u000a surface indicates that the fluorescent color of silkworm cocoons is made up of two peaks of yellow and blue-purple fluorescence\\u000a emission. The fluorescent difference between male and female cocoons is attributed to the differential absorption of yellow\\u000a fluorescent substances by

  14. Construction of transgenic silkworm spinning antibacterial silk with fluorescence.

    PubMed

    Li, Zhen; Jiang, Yue; Cao, Guangli; Li, Jingzhi; Xue, Renyu; Gong, Chengliang

    2015-01-01

    A targeting vector consisting of a fusion gene of the green fluorescent protein (GFP) gene gfp and the antimicrobial peptide cecropin gene cec flanked by pieces of the 5' and 3' sequences of the fibroin L chain gene fib-L of the silkworm (Bombyx mori) and a negative selection DsRed marker gene driven by the baculovirus immediate early gene 1 (i.e.-1) promoter, was used to target the silkworm genome in order to explore the possibility of improving the performance of silk. A transgenic silkworm with a green fluorescent cocoon was obtained and PCR analysis of its genome confirmed that the target genes had been integrated into the silkworm genome correctly. Furthermore, in the posterior silk glands of the G6 generation transformation silkworm, a band representing the fusion protein Fib-L-GFP-Cec with a molecular mass of 68.7 kDa was detected by western blotting with an antibody against GFP. An investigation of the number of bacteria attached to a cocoon showed the transgenic silkworm cocoon possessed antibacterial properties. These results suggested the performance of silk can be improved by modifying the fibroin gene. PMID:25223857

  15. Biological and productive characteristics of silkworm mulberry varieties of different ploidy and their use for raising silkworms in different seasons

    Microsoft Academic Search

    E. F. Leinveber; V. V. Bogoslovskii; M. I. Selionova

    2011-01-01

    The biological and productive characteristics of heteroploid silkworm mulberry varieties of different ploidy are characterized.\\u000a As a result of feeding silkworm caterpillars the leaves of polyploid mulberry varieties compared with the dip0loid variety\\u000a Pc-109, the caterpillar stage is shortened and the viability of caterpillars and palatability and assimilability of leaves,\\u000a cocoon weight, cocoon yield per gram of caterpillars and per

  16. Three pheromone-binding proteins in olfactory sensilla of the two silkmoth species Antheraea polyphemus and Antheraea pernyi.

    PubMed

    Maida, R; Krieger, J; Gebauer, T; Lange, U; Ziegelberger, G

    2000-05-01

    Females of the sibling silkmoth species Antheraea polyphemus and A. pernyi use the same three sex pheromone components in different ratios to attract conspecific males. Accordingly, the sensory hairs on the antennae of males contain three receptor cells sensitive to each of the pheromone components. In agreement with the number of pheromones used, three different pheromone-binding proteins (PBPs) could be identified in pheromone-sensitive hairs of both species by combining biochemical and molecular cloning techniques. MALDI-TOF MS of sensillum lymph droplets from pheromone-sensitive sensilla trichodea of male A. polyphemus revealed the presence of three major peaks with m/z of 15702, 15752 and 15780 and two minor peaks of m/z 15963 and 15983. In Western blots with four antisera raised against different silkmoth odorant-binding proteins, immunoreactivity was found only with an anti-(Apol PBP) serum. Free-flow IEF, ion-exchange chromatography and Western blot analyses revealed at least three anti-(Apol PBP) immunoreactive proteins with pI values between 4.4 and 4.7. N-Terminal sequencing of these three proteins revealed two proteins (Apol PBP1a and Apol PBP1b) identical in the first 49 amino acids to the already known PBP (Apol PBP1) [Raming, K. , Krieger, J. & Breer, H. (1989) FEBS Lett. 256, 2215-2218] and a new PBP having only 57% identity with this amino-acid region. Screening of antennal cDNA libraries with an oligonucleotide probe corresponding to the N-terminal end of the new A. polyphemus PBP, led to the discovery of full length clones encoding this protein in A. polyphemus (Apol PBP3) and in A. pernyi (Aper PBP3). By screening the antennal cDNA library of A. polyphemus with a digoxigenin-labelled A. pernyi PBP2 cDNA [Krieger, J., Raming, K. & Breer, H. (1991) Biochim. Biophys. Acta 1088, 277-284] a homologous PBP (Apol PBP2) was cloned. Binding studies with the two main pheromone components of A. polyphemus and A. pernyi, the (E,Z)-6, 11-hexadecadienyl acetate (AC1) and the (E,Z)-6,11-hexadecadienal (ALD), revealed that in A. polyphemus both Apol PBP1a and the new Apol PBP3 bound the 3H-labelled acetate, whereas no binding of the 3H-labelled aldehyde was found. In A. pernyi two PBPs from sensory hair homogenates showed binding affinity for the AC1 (Aper PBP1) and the ALD (Aper PBP2), respectively. PMID:10806387

  17. Space flight experiment on chinese silkworm on board the russian 10th biosatellite

    Microsoft Academic Search

    Shi Zhizhen; Zhuang Dahuan; Eugene A. Ilyin

    1998-01-01

    Space flight experiments on Chinese silkworm (Bombyx mori L.) were conducted on board the Russian 10th Biosatellite for 12 days. The samples included silkworm eggs, larvae, cocoons, pupae and moths. The processes of spinning, cocooning, mating, oviposition, larval hatching, pupation and moth emergence all completed well in space. The following effects of space flight on silkworm development were observed: The

  18. Use of silkworms for identification of drug candidates having appropriate pharmacokinetics from plant sources

    Microsoft Academic Search

    Yukihiro Asami; Ryo Horie; Hiroshi Hamamoto; Kazuhisa Sekimizu

    2010-01-01

    BACKGROUND: We use silkworms to evaluate therapeutic effects of drug candidates. Our previous reports have revealed that there are common mechanisms of pharmacokinetics of chemicals in silkworms and mammals. In this report, we attempt to establish a method by using silkworms to identify chemicals from plant extracts which are absorbed from intestine and also stably exist in body fluids. RESULTS:

  19. Structure and physical properties of silkworm cocoons.

    PubMed

    Chen, Fujia; Porter, David; Vollrath, Fritz

    2012-09-01

    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

  20. 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

  1. 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%. PMID:11124211

  2. Identification and localization of visual pigments in the retina of the moth, Antheraea polyphemus (Insecta, Saturniidae)

    Microsoft Academic Search

    Helmut Langer; Gudrun Schmeinck; Friederike Anton-Erxleben

    1986-01-01

    In the compound eye of the moth Antheraea polyphemus, three types of visual pigments were found in extracts from the retina and by microspectrophotometry in situ. The absorption maxima of the receptor pigment P and the metarhodopsin M are at (1) P 520–530 nm, M 480–490 nm; (2) P 460–480 nm, M 530–540 nm; (3) P 330–340 nm, M 460–470

  3. Probing a pheromone binding protein of the silkmoth Antheraea polyphemus by endogenous tryptophan fluorescence

    Microsoft Academic Search

    Stefanie Bette; Heinz Breer; Jürgen Krieger

    2002-01-01

    One subtype of the pheromone binding proteins of the silkmoth Antheraea polyphemus (ApolPBP1) has been analysed exploiting the two endogenous tryptophan residues as fluorescent probe. The intrinsic fluorescence exhibited a rather narrow spectrum with a maximum at 336 nm. Site-directed mutagenesis experiments revealed that one of the tryptophan residues (Trp37) is located in a hydrophobic environment whereas Trp127 is more

  4. Resolution of pheromone pulses in receptor cells of Antheraea polyphemus at different temperatures

    Microsoft Academic Search

    B. Kodadová

    1996-01-01

    The ability of pheromone receptor cells of male Antheraea polyphemus (Saturniidae) to resolve stimulus pulses was determined at different temperatures (8°, 18°, 28°C). The cells were stimulated by repeated 20-ms puffs of the pheromone components (E, Z)-6, 11-hexadecadienyl acetate and (E, Z)-6,11-hexadecadienal. At higher temperatures, higher frequencies of stimulus pulses were resolved by the nerve-impulse response: about 1.25 pulses per

  5. A CALCIUM-ACTIVATED NONSPECIFIC CATION CHANNEL FROM OLFACTORY RECEPTOR NEURONES OF THE SILKMOTH ANTHERAEA POLYPHEMUS

    Microsoft Academic Search

    T. A. KEIL

    Summary Single-channel patch-clamp techniques were used to identify and characterize a Ca2+-activated nonspecific cation channel (CAN channel) on insect olfactory receptor neurones (ORNs) from antennae of male Antheraea polyphemus. The CAN channel was found both in acutely isolated ORNs from developing pupae and in membrane vesicles from mature ORNs that presumably originated from inner dendritic segments. Amplitude histograms of the

  6. Kinetic Properties of a Sex Pheromone-Degrading Enzyme: The Sensillar Esterase of Antheraea polyphemus

    Microsoft Academic Search

    Richard G. Vogt; Lynn M. Riddiford; Glenn D. Prestwich

    1985-01-01

    Behavioral and electrophysiological evidence has suggested that sex pheromone is rapidly inactivated within the sensory hairs soon after initiation of the action-potential spike. We report the isolation and characterization of a sex-pheromone-degrading enzyme from the sensory hairs of the silkmoth Antheraea polyphemus. In the presence of this enzyme at physiological concentration, the pheromone [(6E, 11Z)-hexadecadienyl acetate] has an estimated half-life

  7. Female Attacus atlas Respond to Pheromones of Antheraea polyphemus: a Comparative Electrophysiological and Biochemical Study

    Microsoft Academic Search

    Rosario Maida; Jürgen Ziesmann

    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

  8. Rhythms of male Antheraea polyphemus Attraction and female attractiveness, and an improved pheromone sythesis

    Microsoft Academic Search

    J. P. Kochansky; R. T. Cardé; E. F. Taschenberg; W. L. Roelofs

    1977-01-01

    WildAntheraea polyphemus (Cramer) (Lepidoptera:Saturniidae) males were attracted to synthetic pheromone and to laboratory-reared females from 23:00 (EDT) until dawn. Males were attracted to the synthetic pheromone at a mean time of 01:33 and to the females at 03:23 (EDT). Synthetic traps accumulated the most males (61 compared to 19 in the female-baited traps), but there was no large difference in

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

    Microsoft Academic Search

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

    2010-01-01

    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

  10. Mechanism of fluorescent cocoon sex identification for silkworms Bombyx mori.

    PubMed

    Zhang, YuQing; Yu, XiaoHua; Shen, WeiDe; Ma, YongLei; Zhou, LiXia; Xu, NaiXi; Yi, ShuQian

    2010-11-01

    By using silkworms, Bombyx mori, fluorescent cocoon sex identification (FCSI) as an experimental material, direct fluorescence spectrometry of the cocoon surface indicates that the fluorescent color of silkworm cocoons is made up of two peaks of yellow and blue-purple fluorescence emission. The fluorescent difference between male and female cocoons is attributed to the differential absorption of yellow fluorescent substances by the midgut tissue of 5th instar female silkworms. Thin layer chromatography (TLC) and fluorescent spectra indicate that blue-purple fluorescent substances are composed of at least five blue-purple fluorescent pigments, and yellow fluorescent substances are made up of at least three. UV spectra and AlCl? color reaction show that the three fluorescent yellow pigments are flavonoids or their glycosides. Silkworm FCSI is due to selective absorption or accumulation of the yellow fluorescent pigments by the posterior midgut cells of female 5th instar larvae. The cells of the FCSI silkworm midgut, especially the cylinder intestinal cells of the posterior midgut have a component which is a yellow fluorescent pigment-specific binding protein that may be vigorously expressed in the 5th instar larvae. PMID:21046325

  11. [Emergy analysis of mulberry-silkworm ecosystem in China].

    PubMed

    Chen, Mingang; Jin, Peihua; Huang, Lingxia; Lu, Xingmeng

    2006-02-01

    Mulberry-silkworm ecosystem is one of the important agro-ecosystems in China. Based on the principles and methods of emergy analysis, this paper studied the interior structure of mulberry-silkworm ecosystem and its relationship with exterior environment and economy. Some emergy indices for this ecosystem were quantitatively calculated, and compared with those of the agro-ecosystem in China. The results showed that the emergy investment ratio, emergy yield ratio, environmental loading ratio and emergy sustainability index was 3.78, 4.68, 0.18 and 26.0, respectively, suggesting the low environmental pressure and good ecological benefit in mulberry-silkworm ecosystem in China. Hi-technology was required to further decrease the labor force input and enhance the comprehensive utilization of sericultural resources. PMID:16706044

  12. Fruit Body Formation on Silkworm by Cordyceps militaris

    PubMed Central

    Kang, Pil-Don; Kim, Ki-Young; Nam, Sung-Hee; Lee, Man-Young; Choi, Yong-Soo; Kim, Nam-Suk; Kim, Hye-Kyung; Lee, Kwang-Gill; Humber, Richard A.

    2010-01-01

    Injection inoculation protocols for fruit body formation of Cordyceps militaris (C. militaris) were investigated to improve the incidence of infection in the silkworm species Bombyx mori (B. mori). Injection, with suspensions of C. militaris hyphal bodies into living silkworm pupae, was used to test for fruit body production. Use of Daeseungjam rather than Baegokjam or Keumokjam varieties of B. mori is thought to be suitable for infection by C. militaris. From mounting, nine-day-old to 11-day-old pupae showed the best incidence of infection with a 100 µL injection volume. Silkworm pupae injected with a hyphal suspension concentration of more than 2 × 105 colony-forming unit (cfu) recorded a greater than 96% incidence of infection. Also, fruit bodies of C. militaris were induced and produced at a light intensity between 500 and 1,000 lx. PMID:23956640

  13. Horizontal gene transfer in silkworm, Bombyx mori

    PubMed Central

    2011-01-01

    Background The domesticated silkworm, Bombyx mori, is the model insect for the order Lepidoptera, has economically important values, and has gained some representative behavioral characteristics compared to its wild ancestor. The genome of B. mori has been fully sequenced while function analysis of BmChi-h and BmSuc1 genes revealed that horizontal gene transfer (HGT) maybe bestow a clear selective advantage to B. mori. However, the role of HGT in the evolutionary history of B. mori is largely unexplored. In this study, we compare the whole genome of B. mori with those of 382 prokaryotic and eukaryotic species to investigate the potential HGTs. Results Ten candidate HGT events were defined in B. mori by comprehensive sequence analysis using Maximum Likelihood and Bayesian method combining with EST checking. Phylogenetic analysis of the candidate HGT genes suggested that one HGT was plant-to- B. mori transfer while nine were bacteria-to- B. mori transfer. Furthermore, functional analysis based on expression, coexpression and related literature searching revealed that several HGT candidate genes have added important characters, such as resistance to pathogen, to B. mori. Conclusions Results from this study clearly demonstrated that HGTs play an important role in the evolution of B. mori although the number of HGT events in B. mori is in general smaller than those of microbes and other insects. In particular, interdomain HGTs in B. mori may give rise to functional, persistent, and possibly evolutionarily significant new genes. PMID:21595916

  14. Breeding of a Silkworm Variety for Synnemata Production of Isaria tenuipes.

    PubMed

    Kang, Pil-Don; Sung, Gyoo-Byung; Kim, Kee-Young; Kim, Mi-Ja; Hong, In-Pyo; Ha, Nam-Gyu

    2010-09-01

    This study was conducted out to select a silkworm variety suitable for synnemata production of Isaria tenuipes. Four kinds of the mulberry silkworm varieties, Bombyx mori, were hybridized using a Japanese parental line and a Chinese parental line, and used to test for synemata formation in I. tenuipes. The larval period of normal silkworms was 22 hr longer than the silkworms inoculated with this fungus. Among the silkworm varieties tested, Hachojam had the shortest larval period with 23.02 days. The non-cocooning silkworm had a shorter larval period than the cocoon producing silkworms. The pupation rate of normal silkworms was about 9% higher than that of silkworms sprayed with I. tenuipes. Hachojam had the highest infection rate at 99.8%, but no significant difference was observed for the infection rate by silkworm variety. The production of synnemata was the best in JS171 × CS188 with an incidence rate of 99.3%, followed by Hachojam, and Chugangjam. The synnemata produced from Hachojam were the heaviest and showed white or milky-white in color. PMID:23956651

  15. Breeding of a Silkworm Variety for Synnemata Production of Isaria tenuipes

    PubMed Central

    Kang, Pil-Don; Sung, Gyoo-Byung; Kim, Kee-Young; Kim, Mi-Ja; Ha, Nam-Gyu

    2010-01-01

    This study was conducted out to select a silkworm variety suitable for synnemata production of Isaria tenuipes. Four kinds of the mulberry silkworm varieties, Bombyx mori, were hybridized using a Japanese parental line and a Chinese parental line, and used to test for synemata formation in I. tenuipes. The larval period of normal silkworms was 22 hr longer than the silkworms inoculated with this fungus. Among the silkworm varieties tested, Hachojam had the shortest larval period with 23.02 days. The non-cocooning silkworm had a shorter larval period than the cocoon producing silkworms. The pupation rate of normal silkworms was about 9% higher than that of silkworms sprayed with I. tenuipes. Hachojam had the highest infection rate at 99.8%, but no significant difference was observed for the infection rate by silkworm variety. The production of synnemata was the best in JS171 × CS188 with an incidence rate of 99.3%, followed by Hachojam, and Chugangjam. The synnemata produced from Hachojam were the heaviest and showed white or milky-white in color. PMID:23956651

  16. Diabetic silkworms for evaluation of therapeutically effective drugs against type II diabetes

    PubMed Central

    Matsumoto, Yasuhiko; Ishii, Masaki; Hayashi, Yohei; Miyazaki, Shinya; Sugita, Takuya; Sumiya, Eriko; Sekimizu, Kazuhisa

    2015-01-01

    We previously reported that sugar levels in the silkworm hemolymph, i.e., blood, increase immediately (within 1?h) after intake of a high-glucose diet, and that the administration of human insulin decreases elevated hemolymph sugar levels in silkworms. In this hyperglycemic silkworm model, however, administration of pioglitazone or metformin, drugs used clinically for the treatment of type II diabetes, have no effect. Therefore, here we established a silkworm model of type II diabetes for the evaluation of anti-diabetic drugs such as pioglitazone and metformin. Silkworms fed a high-glucose diet over a long time-period (18?h) exhibited a hyperlipidemic phenotype. In these hyperlipidemic silkworms, phosphorylation of JNK, a stress-responsive protein kinase, was enhanced in the fat body, an organ that functionally resembles the mammalian liver and adipose tissue. Fat bodies isolated from hyperlipidemic silkworms exhibited decreased sensitivity to human insulin. The hyperlipidemic silkworms have impaired glucose tolerance, characterized by high fasting hemolymph sugar levels and higher hemolymph sugar levels in a glucose tolerance test. Administration of pioglitazone or metformin improved the glucose tolerance of the hyperlipidemic silkworms. These findings suggest that the hyperlipidemic silkworms are useful for evaluating the hypoglycemic activities of candidate drugs against type II diabetes. PMID:26024298

  17. Diabetic silkworms for evaluation of therapeutically effective drugs against type II diabetes.

    PubMed

    Matsumoto, Yasuhiko; Ishii, Masaki; Hayashi, Yohei; Miyazaki, Shinya; Sugita, Takuya; Sumiya, Eriko; Sekimizu, Kazuhisa

    2015-01-01

    We previously reported that sugar levels in the silkworm hemolymph, i.e., blood, increase immediately (within 1?h) after intake of a high-glucose diet, and that the administration of human insulin decreases elevated hemolymph sugar levels in silkworms. In this hyperglycemic silkworm model, however, administration of pioglitazone or metformin, drugs used clinically for the treatment of type II diabetes, have no effect. Therefore, here we established a silkworm model of type II diabetes for the evaluation of anti-diabetic drugs such as pioglitazone and metformin. Silkworms fed a high-glucose diet over a long time-period (18?h) exhibited a hyperlipidemic phenotype. In these hyperlipidemic silkworms, phosphorylation of JNK, a stress-responsive protein kinase, was enhanced in the fat body, an organ that functionally resembles the mammalian liver and adipose tissue. Fat bodies isolated from hyperlipidemic silkworms exhibited decreased sensitivity to human insulin. The hyperlipidemic silkworms have impaired glucose tolerance, characterized by high fasting hemolymph sugar levels and higher hemolymph sugar levels in a glucose tolerance test. Administration of pioglitazone or metformin improved the glucose tolerance of the hyperlipidemic silkworms. These findings suggest that the hyperlipidemic silkworms are useful for evaluating the hypoglycemic activities of candidate drugs against type II diabetes. PMID:26024298

  18. Species-specific pheromonal compounds induce distinct conformational changes of pheromone binding protein subtypes from Antheraea polyphemus

    Microsoft Academic Search

    Claudia Mohl; Heinz Breer; Jürgen Krieger

    2002-01-01

    .   We have investigated the structural features of three pheromone binding protein (PBP) subtypes from Antheraea polyphemus and monitored possible changes induced upon interaction with the Antheraea pheromonal compounds 4E,9Z-14:Ac [(E4,Z9)-tetradecadienyl-1-acetate], 6E,11Z-16:Ac [(E6,Z11)-hexadecadienyl-1-acetate], and\\u000a 6E,11Z-16:Al [(E6,Z11)-hexadecadienal]. Circular dichroism and second derivative UV-difference spectroscopy data demonstrate\\u000a that the structure of subtype PBP1 significantly changes upon binding of 4E,9Z-14:Ac. The related 6E,11Z-16:Ac

  19. Properties of Xanthine Dehydrogenase in the Silkworm, Bombyx mori L

    Microsoft Academic Search

    Yoshiyuki Hayashi

    1961-01-01

    THE enzyme which catalyses the oxidation of purine derivatives xanthine and hypoxanthine to uric acid has been demonstrated in several insects1-5. In the silkworm, Bombyx mori, it functions as a dehydrogenase. This inference is based on the fact that the formation of uric acid and oxygen consumption of the fat body homogenates show marked increases only when methylene blue is

  20. 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...

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

    Microsoft Academic Search

    Fujia Chen; David Porter; Fritz Vollrath

    2010-01-01

    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

  2. A PRELIMINARY STUDY OF STEAM DRYING OF SILKWORM COCOONS

    Microsoft Academic Search

    Chen Shi-Ruo; Chen Jin-Yong; Arun S. Mujumdar

    1992-01-01

    An experimental study was conducted to explore the feasibility of silkworm cocoon drying with superheated steam. The influence of steam drying on the drying kinetics and the technological characteristics of the dried cocoons is discussed. A promising new technology of cocoon drying is suggested to improve cocoon quality and decrease raw cocoon consumption in silk production.

  3. Transgenic silkworms that weave recombinant proteins into silk cocoons.

    PubMed

    Tomita, Masahiro

    2011-04-01

    As a result of breeding for more than 4,000 years, the silkworm, Bombyx mori, has acquired the ability to synthesize bulk amounts of silk proteins in its silk glands. To utilize this capacity for mass production of useful proteins, transgenic silkworms were generated that synthesized recombinant proteins in the silk gland and secreted them into the silk cocoon. The silk gland is classified into two main regions: the posterior (PSG) and the middle silk gland (MSG). By controlling the expressed regions of the recombinant protein gene in the silk gland, we were able to control the localization of the synthesized protein in the silk thread. Expression in the PSG or MSG led to localization in the insoluble fibroin core or hydrophilic outer sericin layer, respectively. This review focuses on the expression of recombinant protein in the MSG of transgenic silkworms. The recombinant protein secreted in the sericin layer is extractable from the cocoon with only a small amount of endogenous silk protein contamination by soaking the cocoon in mild aqueous solutions. The possibility of utilizing transgenic silkworms as a valuable tool for the mass production of therapeutic and industrially relevant recombinant proteins is discussed. PMID:21184136

  4. Flavonoid 5-glucosides from the cocoon shell of the silkworm, Bombyx mori

    Microsoft Academic Search

    Yasumori Tamura; Ken-ichi Nakajima; Ken-ichi Nagayasu; Chiyuki Takabayashi

    2002-01-01

    The flavonoid 5-glucosides, quercetin 5,4?-di-O-?-d-glucopyranoside and quercetin 5,7,4?-tri-O-?-d-glucopyranoside, together with the known quercetin 5-O-?-d-glucopyranoside, were isolated from the cocoon shell of the silkworm, Bombyx mori. The structures were identified by spectroscopic analysis. These flavonoid glucosides were not present in mulberry leaves, the silkworm's only food, and they are considered to be metabolites produced by the silkworm.

  5. Generation of transgenic silkworms for production of erythropoietin in Bombyx mori

    Microsoft Academic Search

    Jiyeon Seong; Min-Jung Kim; Hyo-Sun Kim; Sun-Ah Kim; Hyung-Wook Jeon; Seung-Hyun Sung; Keun-Chong Kim; Dong Sang Suh

    2011-01-01

    There have been many attempts to generate various essential proteins using transformed E. coli systems. However, prokaryote systems are not equipped with the protein maturation mechanisms necessary to generate eukaryotic\\u000a proteins. In this sense, among the eukaryotes, silkworms have major merits in overcoming the difficulties. Such protein maturation\\u000a mechanisms are available in silkworms. In this study, a transgenic silkworm producing

  6. A receptor and binding protein interplay in the detection of a distinct pheromone component in the silkmoth Antheraea polyphemus

    Microsoft Academic Search

    Maike Forstner; Heinz Breer; Jürgen Krieger

    2009-01-01

    Male moths respond to conspecific female-released pheromones with remarkable sensitivity and specificity, due to highly specialized chemosensory neurons in their antennae. In An- theraea silkmoths, three types of sensory neurons have been described, each responsive to one of three pheromone components. Since also three different pheromone binding pro- teins (PBPs) have been identified, the antenna of Antheraea seems to provide

  7. Morphogenesis of the antenna of the male silkmoth, Antheraea polyphemus. VI. Experimental disturbance of antennal branch formation

    Microsoft Academic Search

    Cornelia Steiner; Thomas A. Keil

    1995-01-01

    In the male silkmoth Antheraea polyphemus, the formation of the side branches of the quadripectinate antennal flagellum was disturbed by an experimental manipulation. Normally the side branches develop in the pupa via deep incisions which proceed from the periphery towards the centerline of the leaf-shaped antennal anlage. Local removal of the uppermost, pigmented cuticular layers of the pupal antennal pocket

  8. Pheromone-binding Proteins Contribute to the Activation of Olfactory Receptor Neurons in the Silkmoths Antheraea polyphemus and Bombyx mori

    Microsoft Academic Search

    Blanka Pophof

    2004-01-01

    The sensilla trichodea of the silkmoth Antheraea polyphemus are innervated by three types of receptor neurons each responding specifically to one of three pheromone components. The sensillum lymph of these sensilla surrounding the sensory dendrites contains three different types of pheromone-binding proteins (PBPs) in high concentrations. The sensilla trichodea of the silkmoth Bombyx mori are supplied by two receptor neurons

  9. Olfactory dendrites of the silkmoth antheraea polyphemus cramer (Lepidoptera : Saturniidae) : immunofluorescent staining of cytoskeletal proteins and transmission electron microscopy

    Microsoft Academic Search

    George L Kumar; Thomas A Keil

    1998-01-01

    In this paper we show the presence of major cytoskeletal proteins : tubulin, globular actin, and intermediate filament-like protein, 2 motor proteins : kinesin and unconventional myosin, and fodrin, in the olfactory dendrites of the silkmoth Antheraea polyphemus (Cramer) (Lepidoptera : Saturniidae) by means of immunofluorescence microscopy. We also show cross-bridges between the membrane and the microtubules and between the

  10. Ultrastructural studies on neuromuscular contacts and the formation of junctions in the flight muscle of Antheraea polyphemus (Lep.)

    Microsoft Academic Search

    H. Nüesch; R. F. Stocker

    1975-01-01

    In the moth Antheraea polyphemus the innervation of the anlage of the dorsolongitudinal flight muscle (dlm) was transected at the onset of adult development. The subsequent breakdown of the isolated motor stumps during early adult development was studied at the ultrastructural level. First reactions are seen on the second day of development when axonal mitochondria shrink. Later, elongated vesicles similar

  11. Morphogenesis of the antenna of the male silkmoth, Antheraea polyphemus. V. Development of the peripheral nervous system

    Microsoft Academic Search

    Cornelia Steiner; Thomas a. Keil

    1995-01-01

    The imaginal antenna of the male silkmoth Antheraea polyphemus is a feather-shaped structure consisting of about 30 flagellomeres, each of which gives off two pairs of side branches. During the pupal stage (lasting for 3 weeks), the antenna develops from a leaf-shaped, flattened epidermal sac (‘antennal blade’) via two series of incisions which proceed from the periphery towards the prospective

  12. Ultrastructural studies on neuromuscular contacts and the formation of junctions in the flight muscle of Antheraea polyphemus (Lep.)

    Microsoft Academic Search

    R. F. Stocker; H. Nüesch

    1975-01-01

    The ultrastructure of neuromuscular connections on developing dorsolongitudinal flight muscles was studied in the moth Antheraea polyphemus, Undifferentiated membrane contacts between axon terminals and muscle-fiber anlagen are present in the diapause pupa. They persist during the period of nerve outgrowth, which probably provides a pathway of contact guidance. By the 4th day of adult development some of these contact areas

  13. Morphogenesis of the antenna of the male silkmoth, Antheraea polyphemus. IV. Segmentation and branch formation.

    PubMed

    Steiner, C; Keil, T A

    1993-06-01

    The imaginal antenna of the male silkmoth Antheraea polyphemus is a featherlike structure; its flagellum consists of about 30 stem segments each giving off two pairs of side branches. The antenna develops during the pupal stage (lasting in total about 21 days) from a leaf-shaped anlage by incisions proceeding from the periphery towards the prospective antennal stem. Primary incisions, starting about 3 days after apolysis, form double branches, which arethen split into single branches by parallel running secondary incisions. The initial pattern of tracheae and peripheral nerves is completely rearranged during these morphogenetic processes which are finished 9-10 days after apolysis. In Antheraea the dorsal and ventral epithelial monolayers of the antennal anlage are successively subdivided during development into a pattern of repetitive epithelial zones. Within the first day after apolysis alternating stripes of sensillogenic and non-sensillogenic epithelium are differentiating. Then the latter are further subdivided, and at last four different stripelike zones (I-IV) can be discriminated. Long basal protrusions of the epidermal cells ('epidermal feet'), and most probably haemocytes, seem to be involved in the reconstruction of the epithelium: both show characteristic arrangements within the antennal anlage during successive developmental stages. PMID:18621244

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

    Microsoft Academic Search

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

    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

  15. 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 statuses are largely different (eating F=3.191, P<0.05; non-eating F=2.935, P <0.05). Moreover, the amounts of O2 inhaled and CO2 exhaled by the silkworms in the first three developing stages were much more than those of the silkworms in the latter two developing stages per unit weight of leaves. These results concerning the nutrient compositions and respiration characteristics of silkworms can provide valuable data for the establishment of complex bioregenerative life support systems including different biological units in the lunar or mars bases in the future.

  16. Oak Poisoning in Livestock. 

    E-print Network

    Dollahite, J. W.; Housholder, G. T.; Camp, B. J.

    1966-01-01

    April 1966 r Oak Poisoning *. . TEXAS A&M UNIVERSITY Texas Agricultural Experiment Station R. E. Patterson, Director, College Station, Texas Summary Oak poisoning is a major problem in the production of livestock in areas where oak occurs.... The blossoms, buds, young leaves and acorns are poisonous. Cattle, sheep, goats, swine, rabbits and guinea pigs are susceptible to oak poisoning. A gallotannin isolated from oak has been demonstrated to be poisonous. Calcium hydroxide is an antidote...

  17. Silkworm and spider silk scaffolds for chondrocyte support

    Microsoft Academic Search

    Kris Gellynck; Peter C. M. Verdonk; Els Van Nimmen; Karl F. Almqvist; Tom Gheysens; Gustaaf Schoukens; Lieva Van Langenhove; Paul Kiekens; Johan Mertens; Gust Verbruggen

    2008-01-01

    Objective To create scaffolds with silkworm cocoon, spider egg sac and spider dragline silk fibres and examine their use for chondrocyte\\u000a attachment and support. Methods Three different kinds of scaffolds were developed with Bombyx mori cocoon, Araneus diadematus egg sac and dragline silk fibres. The attachment of human articular cartilage cells were investigated on these bioprotein\\u000a matrices. The chondrocytes produced

  18. 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.

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

    Microsoft Academic Search

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

    2008-01-01

    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

  20. Identification and Characterization of an Arginine Kinase as a Major Allergen from Silkworm (Bombyx mori) Larvae

    Microsoft Academic Search

    Zhigang Liu; Lixin Xia; Yulan Wu; Qingyou Xia; Jiajie Chen; Kenneth H. Roux

    2009-01-01

    Background: The silkworm, Bombyx mori, is an important insect in the textile industry and its pupa are used in Chinese cuisine and traditional Chinese medicine. The silk, urine and dander of silkworms is often the cause of allergies in sericulture workers and the pupa has been found to be a food allergen in China. Recent studies have focused on reporting

  1. in silico identification of protein-protein interactions in Silkworm, Bombyx mori

    PubMed Central

    Sumathy, Ramasamy; Rao, Ashwath Southekal Krishna; Chandrakanth, Nalavadi; Gopalakrishnan, Velliyur Kanniappan

    2014-01-01

    The Domesticated silkworm, Bombyx mori, an economically important insect has been used as a lepidopteran molecular model next only to Drosophila. Compared to the genomic information in silkworm, the protein-protein interaction data are limited. Therefore experimentally identified PPI maps from five model organisms such as E.coli, C.elegans, D.melanogaster, H. sapiens, S. cerevisiae were used to infer the PPI network of silkworm using the well-recognized Interlog based method. Among the 14623 silkworm proteins, 7736 protein-protein interaction pairs were predicted which include 2700 unique proteins of the silkworms. Using the iPfam interaction domains and the gene expression data, these predictions were validated. In that 625 PPI pairs of predicted network were associated with the iPfam domain-domain interactions and the random network has average of 9. In the gene expression method, the average PCC value of the predicted network and random network was 0.29 and 0.23100±0.00042 respectively. It reveals that the predicted PPI networks of silkworm are highly significant and reliable. This is the first PPI network for the silkworm which will provide a framework for deciphering the cellular processes governing key metabolic pathways in the silkworm, Bombyx mori and available at SilkPPI (http://210.212.197.30/SilkPPI/). PMID:24616555

  2. in silico identification of protein-protein interactions in Silkworm, Bombyx mori.

    PubMed

    Sumathy, Ramasamy; Rao, Ashwath Southekal Krishna; Chandrakanth, Nalavadi; Gopalakrishnan, Velliyur Kanniappan

    2014-01-01

    The Domesticated silkworm, Bombyx mori, an economically important insect has been used as a lepidopteran molecular model next only to Drosophila. Compared to the genomic information in silkworm, the protein-protein interaction data are limited. Therefore experimentally identified PPI maps from five model organisms such as E.coli, C.elegans, D.melanogaster, H. sapiens, S. cerevisiae were used to infer the PPI network of silkworm using the well-recognized Interlog based method. Among the 14623 silkworm proteins, 7736 protein-protein interaction pairs were predicted which include 2700 unique proteins of the silkworms. Using the iPfam interaction domains and the gene expression data, these predictions were validated. In that 625 PPI pairs of predicted network were associated with the iPfam domain-domain interactions and the random network has average of 9. In the gene expression method, the average PCC value of the predicted network and random network was 0.29 and 0.23100±0.00042 respectively. It reveals that the predicted PPI networks of silkworm are highly significant and reliable. This is the first PPI network for the silkworm which will provide a framework for deciphering the cellular processes governing key metabolic pathways in the silkworm, Bombyx mori and available at SilkPPI (http://210.212.197.30/SilkPPI/). PMID:24616555

  3. Identification of a New Protein from Silkworm Pupas by Biological Mass Spectrometry

    Microsoft Academic Search

    Liang Jiang; Qiong Liu; Decong Hu; Huibi Xu; Hongxia Wang

    2005-01-01

    To find out the efficient peptide of selenium-rich silkworm pupas in inducing the apoptosis of hepatoma cells, selenium-containing proteins were isolated and characterized. One of the two major proteins in selenium-rich silkworm pupas was identified to be a new protein by peptide mass fingerprinting on matrix assisted laser desorption ionization - time of flight - mass spectrometry. Amino acid sequences

  4. A draft sequence for the genome of the domesticated silkworm (Bombyx mori).

    PubMed

    Xia, Qingyou; Zhou, Zeyang; Lu, Cheng; Cheng, Daojun; Dai, Fangyin; Li, Bin; Zhao, Ping; Zha, Xingfu; Cheng, Tingcai; Chai, Chunli; Pan, Guoqing; Xu, Jinshan; Liu, Chun; Lin, Ying; Qian, Jifeng; Hou, Yong; Wu, Zhengli; Li, Guanrong; Pan, Minhui; Li, Chunfeng; Shen, Yihong; Lan, Xiqian; Yuan, Lianwei; Li, Tian; Xu, Hanfu; Yang, Guangwei; Wan, Yongji; Zhu, Yong; Yu, Maode; Shen, Weide; Wu, Dayang; Xiang, Zhonghuai; Yu, Jun; Wang, Jun; Li, Ruiqiang; Shi, Jianping; Li, Heng; Li, Guangyuan; Su, Jianning; Wang, Xiaoling; Li, Guoqing; Zhang, Zengjin; Wu, Qingfa; Li, Jun; Zhang, Qingpeng; Wei, Ning; Xu, Jianzhe; Sun, Haibo; Dong, Le; Liu, Dongyuan; Zhao, Shengli; Zhao, Xiaolan; Meng, Qingshun; Lan, Fengdi; Huang, Xiangang; Li, Yuanzhe; Fang, Lin; Li, Changfeng; Li, Dawei; Sun, Yongqiao; Zhang, Zhenpeng; Yang, Zheng; Huang, Yanqing; Xi, Yan; Qi, Qiuhui; He, Dandan; Huang, Haiyan; Zhang, Xiaowei; Wang, Zhiqiang; Li, Wenjie; Cao, Yuzhu; Yu, Yingpu; Yu, Hong; Li, Jinhong; Ye, Jiehua; Chen, Huan; Zhou, Yan; Liu, Bin; Wang, Jing; Ye, Jia; Ji, Hai; Li, Shengting; Ni, Peixiang; Zhang, Jianguo; Zhang, Yong; Zheng, Hongkun; Mao, Bingyu; Wang, Wen; Ye, Chen; Li, Songgang; Wang, Jian; Wong, Gane Ka-Shu; Yang, Huanming

    2004-12-10

    We report a draft sequence for the genome of the domesticated silkworm (Bombyx mori), covering 90.9% of all known silkworm genes. Our estimated gene count is 18,510, which exceeds the 13,379 genes reported for Drosophila melanogaster. Comparative analyses to fruitfly, mosquito, spider, and butterfly reveal both similarities and differences in gene content. PMID:15591204

  5. A Draft Sequence for the Genome of the Domesticated Silkworm (Bombyx mori)

    Microsoft Academic Search

    Qingyou Xia; Zeyang Zhou; Cheng Lu; Daojun Cheng; Fangyin Dai; Bin Li; Ping Zhao; Xingfu Zha; Tingcai Cheng; Chunli Chai; Guoqing Pan; Jinshan Xu; Chun Liu; Ying Lin; Jifeng Qian; Yong Hou; Zhengli Wu; Guanrong Li; Minhui Pan; Chunfeng Li; Yihong Shen; Xiqian Lan; Lianwei Yuan; Tian Li; Hanfu Xu; Guangwei Yang; Yongji Wan; Yong Zhu; Maode Yu; Weide Shen; Dayang Wu; Zhonghuai Xiang; Jun Yu; Jun Wang; Ruiqiang Li; Jianping Shi; Heng Li; Guangyuan Li; Jianning Su; Xiaoling Wang; Guoqing Li; Zengjin Zhang; Qingfa Wu; Jun Li; Qingpeng Zhang; Ning Wei; Jianzhe Xu; Haibo Sun; Le Dong; Dongyuan Liu; Shengli Zhao; Xiaolan Zhao; Qingshun Meng; Fengdi Lan; Xiangang Huang; Yuanzhe Li; Lin Fang; Changfeng Li; Dawei Li; Yongqiao Sun; Zhenpeng Zhang; Zheng Yang; Yanqing Huang; Yan Xi; Qiuhui Qi; Dandan He; Haiyan Huang; Xiaowei Zhang; Zhiqiang Wang; Wenjie Li; Yuzhu Cao; Yingpu Yu; Hong Yu; Jinhong Li; Jiehua Ye; Huan Chen; Yan Zhou; Bin Liu; Jing Wang; Jia Ye; Hai Ji; Shengting Li; Peixiang Ni; Jianguo Zhang; Yong Zhang; Hongkun Zheng; Bingyu Mao; Wen Wang; Chen Ye; Songgang Li; Jian Wang; Gane Ka-Shu Wong; Huanming Yang

    2004-01-01

    We report a draft sequence for the genome of the domesticated silkworm (Bombyx mori), covering 90.9% of all known silkworm genes. Our estimated gene count is 18,510, which exceeds the 13,379 genes reported for Drosophila melanogaster. Comparative analyses to fruitfly, mosquito, spider, and butterfly reveal both similarities and differences in gene content.

  6. Survey and analysis of microsatellites in the silkworm, Bombyx mori: frequency, distribution, mutations, marker potential and their conservation in heterologous species.

    PubMed

    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

  7. Marker-assisted selection in breeding silkworm strains with high silk production and resistance to the densonucleosis virus.

    PubMed

    Hou, C X; Sun, P J; Guo, X J; Huang, Y P; Li, M W

    2013-01-01

    In the silkworm (Bombyx mori), resistance to the Zhenjiang (China) strain of the densonucleosis virus (DNV-Z) is controlled by the recessive gene nsd-Z (non-susceptible to DNV-Z), which is linked to 7 simple-sequence repeat markers. Marker-assisted evaluation and selection of DNV-Z-resistant silkworms were used for predicting DNV-resistance in backcrossed animals. A silkworm race was bred using this method, and its economic characteristics were found to be similar to those of commercial silkworm races. These markers will therefore be useful for silkworm breeding programs and in screening for densonucleosis resistance in segregating populations. PMID:24114212

  8. Esterase activity in the olfactory sensilla of the silkmoth Antheraea polyphemus.

    PubMed

    Maida, R; Ziegelberger, G; Kaissling, K E

    1995-03-27

    We studied in individual males of Antheraea polyphemus the activity of the sensillar esterase, a pheromone-degrading enzyme present in the sensillum lymph surrounding the olfactory receptor cells. In parallel, receptor potentials from single pheromone-sensitive sensilla trichodea were recorded. Our screening revealed a large variability of the enzyme activity in individuals with similar electrophysiological responses. In some moths the sensillar esterase was not detectable, i.e. present with 100-fold less activity. However, such variable esterase activity showed no correlation to the time course of the receptor potential. Thus, enzymatic pheromone degradation does not seem to be involved in the rapid pheromone inactivation at the end of the stimulus, but rather serves as the final pheromone sequestration step. PMID:7605955

  9. Identification of cytoskeletal proteins in the antennae of the silkmoths Antheraea polyphemus and A. pernyi.

    PubMed

    Kumar, G L; Maida, R; Keil, T A

    1996-08-12

    Although the overall cytoskeletal morphology of the olfactory dendrite in the antennae of the silkmoths Antheraea polyphemus and A. pernyi is known, the cytoskeleton proteins that structurally and functionally support these structures remain to be identified in this paper, we describe the identification of tubulin, actin and intermediate filament-like proteins in the olfactory dendrites, and motor proteins such as kinesin and unconventional myosin in the antennal branches by the use of antibodies. We also show that the tubulins within the olfactory dendrites and in the antennal branches are acetylated. This study provides valuable information concerning the possible role of these proteins in transduction, transport and motility, as is evident in other systems. PMID:8905709

  10. Identification of PLC beta and PKC in pheromone receptor neurons of Antheraea polyphemus.

    PubMed

    Maida, R; Redkozubov, A; Ziegelberger, G

    2000-06-01

    Two proteins of the IP3 transduction pathway were identified by Western blots in homogenates of isolated pheromone-sensitive sensilla of the silkmoth Antheraea polyphemus. A 110 kDa protein was recognized by an antiserum raised against the Drosophila phospholipase C beta (PLC beta p121) and a 80kDa protein was labelled by an antiserum against a synthetic peptide of a conserved region of protein kinase C (PKC). Incubation of homogenized sensory hairs with the main sex pheromone component, (E,Z) 6-11 hexadecadienyl acetate, resulted in a 6-fold increase in the activity of PKC compared to controls without pheromone. In contrast, incubation with pheromone did not affect the activity of protein kinase A (PKA). Activation of PKC by the membrane permeable dioctanoylglycerol led to excitation of the pheromone-sensitive receptor neurons. These data support the current concept that pheromone perception of moths is mediated by the IP3 transduction pathway. PMID:10852242

  11. Kinetic properties of a sex pheromone-degrading enzyme: the sensillar esterase of Antheraea polyphemus.

    PubMed

    Vogt, R G; Riddiford, L M; Prestwich, G D

    1985-12-01

    Behavioral and electrophysiological evidence has suggested that sex pheromone is rapidly inactivated within the sensory hairs soon after initiation of the action-potential spike. We report the isolation and characterization of a sex-pheromone-degrading enzyme from the sensory hairs of the silkmoth Antheraea polyphemus. In the presence of this enzyme at physiological concentration, the pheromone [(6E,11Z)-hexadecadienyl acetate] has an estimated half-life of 15 msec. Our findings suggest a molecular model for pheromone reception in which a previously reported pheromone-binding protein acts as a pheromone carrier, and an enzyme acts as a rapid pheromone inactivator, maintaining a low stimulus noise level within the sensory hairs. PMID:3001718

  12. Hormonal control of muscle atrophy and degeneration in the moth Antheraea polyphemus.

    PubMed

    Schwartz, L M; Truman, J W

    1984-07-01

    The intersegmental muscles of the giant silkmoth Antheraea polyphemus (Cramer) can undergo two forms of degenerative changes: a wasting atrophy that lasts about 6 days or rapid dissolution that is completed within 30 h. Muscle atrophy is induced by a dramatic decline in the endogenous titres of the steroid moulting hormone 20-hydroxyecdysone. 20-Hydroxyecdysone appears to act as a trophic factor for the muscles as infusion or injection of this steroid blocks further atrophy of the muscle. The normal decline of 20-hydroxyecdysone also allows the muscles to become competent to respond to the peptide eclosion hormone. Eclosion hormone is then released and acts directly on these muscles to induce rapid cell death which is morphologically and physiologically distinct from steroid-regulated atrophy. PMID:6491588

  13. Mineralization and biocompatibility of Antheraea pernyi (A. pernyi) silk sericin film for potential bone tissue engineering.

    PubMed

    Yang, Mingying; Mandal, Namita; Shuai, Yajun; Zhou, Guanshan; Min, Sijia; Zhu, Liangjun

    2014-01-01

    This study aimed to investigate the mineralization of Antheraea pernyi (A. pernyi) silk sericin. Mineralization of A. pernyi sericin was performed by alternative soaking in calcium and phosphate. The inhibition of precipitation of calcium carbonate and von Kossa staining on A. pernyi sericin were tested, and the corresponding results prove that A. pernyi sericin has Ca binding activity. Scanning electron microscope (SEM) observation shows that spherical crystals could be nucleated on the A. pernyi sericin film. These crystals were confirmed to be hydroxyapatite according to FT-IR and XRD spectra, indicating that A. pernyi sericin is capable of mineralization. In addition, cell adhesion and growth activity assay demonstrate that A. pernyi sericin shows excellent biocompatibility for the growth of MG-63 cells. PMID:24211968

  14. MEASURED BEHAVIOURAL LATENCY IN RESPONSE TO SEX-PHEROMONE LOSS IN THE LARGE SILK MOTH ANTHERAEA POLYPHEMUS

    Microsoft Academic Search

    T. C. BAKER

    1988-01-01

    Summary Males of the giant silk moth Antheraea polyphemus Cramer (Lepidoptera: Saturniidae) were video-recorded in a sustained-flight wind tunnel in a constant plume of sex pheromone. The plume was experimentally truncated, and the moths, on losing pheromone stimulus, rapidly changed their behaviour from up- tunnel zig-zag flight to lateral casting flight. The latency of this change was in the range

  15. Binding and hydrolysis of radiolabeled pheromone and several analogs by male-specific antennal proteins of the moth Antheraea polyphemus

    Microsoft Academic Search

    Glenn D. Prestwich; Richard G. Vogt; Lynn M. Riddiford

    1986-01-01

    Sensory hair proteins from antennae of males of the wild silk moth,Antheraea polyphemus (Lepidoptera, Saturniidae) were incubated with radiolabeled 6E,11Z-hexadecadienyl acetate in the presence of unlabeled pheromone analogs as competitive inhibitors. The two extracellular proteins of importance, a highly active sensillar esterase and an abundant 15,000 mol wt binding protein, interact to degrade labeled pheromone less efficiently in the presence

  16. Dynamic Rearrangement Within the Antheraea pernyi Silk Fibroin Gene Is Associated with Four Types of Repetitive Units

    Microsoft Academic Search

    Hideki Sezutsu; Kenji Yukuhiro

    2000-01-01

    .   We characterized a full-length gene encoding wild silkmoth Antheraea pernyi fibroin (Ap-fibroin) to clarify the conformation of repetitive sequences. The gene consisted of a first exon encoding 14\\u000a amino acid residues, a short intron (120 bp), and a long second exon encoding 2,625 amino acid residues. Three amino acids,\\u000a alanine, glycine, and serine, amounted to 81% of the Ap-fibroin

  17. Porphyromonas gingivalis Peptidoglycans Induce Excessive Activation of the Innate Immune System in Silkworm Larvae*

    PubMed Central

    Ishii, Kenichi; Hamamoto, Hiroshi; Imamura, Katsutoshi; Adachi, Tatsuo; Shoji, Mikio; Nakayama, Koji; Sekimizu, Kazuhisa

    2010-01-01

    Porphyromonas gingivalis, a pathogen that causes inflammation in human periodontal tissue, killed silkworm (Bombyx mori, Lepidoptera) larvae when injected into the blood (hemolymph). Silkworm lethality was not rescued by antibiotic treatment, and heat-killed bacteria were also lethal. Heat-killed bacteria of mutant P. gingivalis strains lacking virulence factors also killed silkworms. Silkworms died after injection of peptidoglycans purified from P. gingivalis (pPG), and pPG toxicity was blocked by treatment with mutanolysin, a peptidoglycan-degrading enzyme. pPG induced silkworm hemolymph melanization at the same dose as that required to kill the animal. pPG injection increased caspase activity in silkworm tissues. pPG-induced silkworm death was delayed by injecting melanization-inhibiting reagents (a serine protease inhibitor and 1-phenyl-2-thiourea), antioxidants (N-acetyl-l-cysteine, glutathione, and catalase), and a caspase inhibitor (Ac-DEVD-CHO). Thus, pPG induces excessive activation of the innate immune response, which leads to the generation of reactive oxygen species and apoptotic cell death in the host tissue. PMID:20702417

  18. An amplified fragment length polymorphism map of the silkworm.

    PubMed

    Tan, Y D; Wan, C; Zhu, Y; Lu, C; Xiang, Z; Deng, H W

    2001-03-01

    The silkworm (Bombyx mori L.) is a lepidopteran insect with a long history of significant agricultural value. We have constructed the first amplified fragment length polymorphism (AFLP) genetic linkage map of the silkworm B. mori at a LOD score of 2.5. The mapping AFLP markers were genotyped in 47 progeny from a backcross population of the cross no. 782 x od100. A total of 1248 (60.7%) polymorphic AFLP markers were detected with 35 PstI/TaqI primer combinations. Each of the primer combinations generated an average of 35.7 polymorphic AFLP markers. A total of 545 (44%) polymorphic markers are consistent with the expected segregation ratio of 1:1 at the significance level of P = 0.05. Of the 545 polymorphic markers, 356 were assigned to 30 linkage groups. The number of markers on linkage groups ranged from 4 to 36. There were 21 major linkage groups with 7-36 markers and 9 relatively small linkage groups with 4-6 markers. The 30 linkage groups varied in length from 37.4 to 691.0 cM. The total length of this AFLP linkage map was 6512 cM. Genetic distances between two neighboring markers on the same linkage group ranged from 0.2 to 47 cM with an average of 18.2 cM. The sex-linked gene od was located between the markers P1T3B40 and P3T3B27 at the end of group 3, indicating that AFLP linkage group 3 was the Z (sex) chromosome. This work provides an essential basic map for constructing a denser linkage map and for mapping genes underlying agronomically important traits in the silkworm B. mori L. PMID:11238411

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

    SciTech Connect

    Chen Fujia; Porter, David; Vollrath, Fritz [Department of Zoology, University of Oxford, Oxford OX1 3PS (United Kingdom)

    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.

  20. 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.

  1. Bmovo-1 Regulates Ovary Size in the Silkworm, Bombyx mori

    PubMed Central

    Cao, Guangli; Huang, Moli; Xue, Gaoxu; Qian, Ying; Song, Zuowei; Gong, Chengliang

    2014-01-01

    The regulation of antagonistic OVO isoforms is critical for germline formation and differentiation in Drosophila. However, little is known about genes related to ovary development. In this study, we cloned the Bombyx mori ovo gene and investigated its four alternatively spliced isoforms. BmOVO-1, BmOVO-2 and BmOVO-3 all had four C2H2 type zinc fingers, but differed at the N-terminal ends, while BmOVO-4 had a single zinc finger. Bmovo-1, Bmovo-2 and Bmovo-4 showed the highest levels of mRNA in ovaries, while Bmovo-3 was primarily expressed in testes. The mRNA expression pattern suggested that Bmovo expression was related to ovary development. RNAi and transgenic techniques were used to analyze the biological function of Bmovo. The results showed that when the Bmovo gene was downregulated, oviposition number decreased. Upregulation of Bmovo-1 in the gonads of transgenic silkworms increased oviposition number and elevated the trehalose contents of hemolymph and ovaries. We concluded that Bmovo-1 was involved in protein synthesis, contributing to the development of ovaries and oviposition number in silkworms. PMID:25119438

  2. Production of scFv-conjugated affinity silk powder by transgenic silkworm technology.

    PubMed

    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

  3. Transgenic silkworms (Bombyx mori) produce recombinant spider dragline silk in cocoons.

    PubMed

    Wen, Hongxiu; Lan, Xiqian; Zhang, Yuansong; Zhao, Tianfu; Wang, Yujun; Kajiura, Zenta; Nakagaki, Masao

    2010-04-01

    Spider dragline silk is a unique fibrous protein with a combination of tensile strength and elasticity, but the isolation of large amounts of silk from spiders is not feasible. In this study, we generated germline-transgenic silkworms (Bombyx mori) that spun cocoons containing recombinant spider silk. A piggyBac-based transformation vector was constructed that carried spider dragline silk (MaSp1) cDNA driven by the sericin 1 promoter. Silkworm eggs were injected with the vector, producing transgenic silkworms displaying DsRed fluorescence in their eyes. Genotyping analysis confirmed the integration of the MaSp1 gene into the genome of the transgenic silkworms, and silk protein analysis revealed its expression and secretion in the cocoon. Compared with wild-type silk, the recombinant silk displayed a higher tensile strength and elasticity. The results indicate the potential for producing recombinant spider silk in transgenic B. mori. PMID:19633923

  4. New and highly efficient expression systems for expressing selectively foreign protein in the silk glands of transgenic silkworm

    Microsoft Academic Search

    Aichun Zhao; Tianfu Zhao; Yuansong Zhang; Qingyou Xia; Cheng Lu; Zeyang Zhou; Zhonghuai Xiang; M. Nakagaki

    2010-01-01

    We constructed three different fibroin H-chain expression systems to estimate the efficacy of producing recombinant proteins\\u000a in the cocoon of transgenic silkworms. The results showed that the three different EGFP\\/H-chain fusion genes were all expressed\\u000a selectively in the posterior silk gland of the transgenic silkworm. The recombinant protein content of transgenic silkworm\\u000a cocoons is up to 15% (w\\/w) when using

  5. A germline transgenic silkworm that secretes recombinant proteins in the sericin layer of cocoon

    Microsoft Academic Search

    Masahiro Tomita; Rika Hino; Shingo Ogawa; Masashi Iizuka; Takahiro Adachi; Katsuhiko Shimizu; Hisaya Sotoshiro; Katsutoshi Yoshizato

    2007-01-01

    A silk thread of the silkworm, Bombyx mori, is composed of the insoluble inner fibroin and the hydrophilic outer sericin layer, which are synthesized in the posterior\\u000a and middle silk gland (MSG), respectively. This study aimed to develop a novel sericin 1 gene (ser1) promoter-driven recombinant expression system using transgenic silkworms, in which recombinant proteins are synthesized\\u000a in MSG and

  6. C-prolinylquercetins from the yellow cocoon shell of the silkworm, Bombyx mori

    Microsoft Academic Search

    Chikara Hirayama; Hiroshi Ono; Yasumori Tamura; Masatoshi Nakamura

    2006-01-01

    Two flavonoids containing the l-proline moiety, 6-C-[(2S,5S)-prolin-5-yl] quercetin (prolinalin A) and 6-C-[(2S,5R)-prolin-5-yl] quercetin (prolinalin B), were isolated from the cocoon shell of the silkworm, Bombyx mori. Their structural elucidation was achieved by application of acid hydrolysis and spectroscopic methods. These compounds were not found in the leaves of mulberry (Morus alba L.), the host plant of the silkworm, suggesting that

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

    Microsoft Academic Search

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

    2009-01-01

    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 (42–52goil\\/gsorbent for

  8. Purification and characterization of two major selenium-containing proteins in selenium-rich silkworm pupas

    Microsoft Academic Search

    Qiong Liu; Xueying Liang; Decong Hu; Ping Chen; Jing Tian; Hong Zhang

    2010-01-01

    Selenium (Se) is an essential trace element in vivo involved in the defense against oxidative stress. Se deficiency is associated with many human diseases. The bioactivity of\\u000a Se is dose- and species-dependent. Silkworm pupa has been reported to accumulate Se mainly in proteins. Thus the characterization\\u000a of major Se-containing proteins is very important in the application of Se-rich silkworm pupas

  9. Mechanism of Enhanced Bombyx mori Nucleopolyhedrovirus-Resistance by Titanium Dioxide Nanoparticles in Silkworm

    PubMed Central

    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. PMID:25692869

  10. Characterization of four pupal wing cuticular protein genes of the silkmoth Antheraea polyphemus.

    PubMed

    Kumar, M N; Sridhara, S

    1994-03-01

    Three different clones have been isolated from a genomic library of the silkmoth Antheraea polyphemus by employing a subtractive hybridization technique. The clones with inserts of 13-16 kb of DNA each, code for mRNAs expressed in the wing epidermis during JH induced second pupal cuticle deposition. While two of the clones code for a single mRNA each, the third one codes for two mRNAs. All the four mRNAs code for distinct polypeptides that can be precipitated with antibodies raised against pupal cuticular proteins. These genes are activated at the same period of pupal development and their transcripts follow similar patterns of accumulation. Although these genes are expressed in a tissue and time specific manner attesting to their pupal wing epidermal specificity, three of them are expressed in the adult wing epidermis also, but not at the larval stage. While DNAs from other silkmoths and insects hybridize to these genes, only one of the A. polyphemus genes hybridizes to RNA from second pupal wings of two other silkmoths tested. PMID:7517270

  11. Redox-shift of the pheromone-binding protein in the silkmoth Antheraea polyphemus.

    PubMed

    Ziegelberger, G

    1995-09-15

    In pheromone-sensitive hairs of the male silkmoth Antheraea polyphemus, two electrophoretically distinct pheromone-binding proteins (PBPs) are present. They indicate no amino acid sequence diversity according to peptide mapping, but differ in their redox state, as shown by free-sulfhydryl-group-specific cleavage at cysteine residues with 2-nitro-5-thiocyanobenzoic acid. In kinetic studies, the pheromone was initially bound mainly by the reduced PBP but later by the oxidized PBP, where all six cysteine residues form disulfide bonds. This redox shift was observed only in the homogenate of isolated olfactory hairs, where proteins of the sensillum lymph and receptive dendrites are present. In control experiments with purified binding proteins, the proportion of pheromone bound to the oxidized PBP did not increase with increasing incubation time, suggesting that disulfide formation does not occur spontaneously but is mediated by the sensory hairs, possibly by interaction with the receptor cell membrane. These data suggest that arriving hydrophobic pheromone molecules are first bound by the reduced PBP and transported through the aqueous sensillum lymph towards the receptor molecules of the dendritic membrane. The oxidized complex might not be able to activate further receptors and, thus, effectively deactivate the pheromone molecules within the sensillum lymph. PMID:7588707

  12. Probing a pheromone binding protein of the silkmoth Antheraea polyphemus by endogenous tryptophan fluorescence.

    PubMed

    Bette, Stefanie; Breer, Heinz; Krieger, Jürgen

    2002-03-01

    One subtype of the pheromone binding proteins of the silkmoth Antheraea polyphemus (ApolPBP1) has been analysed exploiting the two endogenous tryptophan residues as fluorescent probe. The intrinsic fluorescence exhibited a rather narrow spectrum with a maximum at 336 nm. Site-directed mutagenesis experiments revealed that one of the tryptophan residues (Trp37) is located in a hydrophobic environment whereas Trp127 is more solvent exposed, as was predicted modeling the ApolPBP1 sequence on the proposed structure of the Bombyx mori pheromone binding protein. Monitoring the interaction of ApolPBP1 as well as its Trp mutants with the three species-specific pheromone compounds by recording the endogenous fluorescence emission revealed profound differences; whereas (E6,Z11)-hexadecadienal induced a dose-dependent quenching of the fluorescence, both (E6,Z11)-hexadecadienyl-1-acetate and (E4,Z9)-tetradecadienyl-1-acetate elicited an augmentation of the endogenous fluorescence. These data indicate that although ApolPBP1 can bind all three pheromones, there are substantial differences concerning their interaction with the protein, which may have important functional implications. PMID:11804795

  13. A calcium-activated nonspecific cation channel from olfactory receptor neurones of the silkmoth Antheraea polyphemus.

    PubMed

    Zufall, F; Hatt, H

    1991-11-01

    Single-channel patch-clamp techniques were used to identify and characterize a Ca2(+)-activated nonspecific cation channel (CAN channel) on insect olfactory receptor neurones (ORNs) from antennae of male Antheraea polyphemus. The CAN channel was found both in acutely isolated ORNs from developing pupae and in membrane vesicles from mature ORNs that presumably originated from inner dendritic segments. Amplitude histograms of the CAN single-channel currents presented well-defined peaks corresponding to at least four channel substates each having a conductance of about 16 pS. Simultaneous gating of the substates was achieved by intracellular Ca2(+) with an EC(50) value of about 80 nmol(-l). Activity of the CAN channel could be blocked by application of amiloride (IC(50)<100 nmoll(-1)). Moreover, in the presence of l ?moll(-1) Ca2+,opening of the CAN channel was totally suppressed by 10 ?moll(-1) cyclic GMP,whereas ATP (1 mmoll(-1)) was without effect. We suggest that the CAN channel plays a specific role in modulation of cell excitability and in shaping the voltage response of ORNs. PMID:22141156

  14. Antennal-specific pheromone-degrading aldehyde oxidases from the moths Antheraea polyphemus and Bombyx mori.

    PubMed

    Rybczynski, R; Vogt, R G; Lerner, M R

    1990-11-15

    Female moths produce blends of odorant chemicals, called pheromones. These precise chemical mixtures both attract males and elicit appropriate mating behaviors. To locate females, male moths must rapidly detect changes in environmental pheromone concentration. Therefore, the regulation of pheromone concentration within antennae, their chief organ of smell, is important. We describe antennal-specific aldehyde oxidases from the moths Antheraea polyphemus and Bombyx mori that are capable of catabolizing long chain, unsaturated aldehydes such as their aldehyde pheromones. These soluble enzymes are associated uniquely with male and female antennae and have molecular masses of 175 and 130 kDa, respectively. The A. polyphemus aldehyde oxidase has been localized to the olfactory sensilla which contain the pheromone receptor cell dendrites. These same sensilla contain a previously described sensilla-specific esterase that degrades the acetate ester component of A. polyphemus pheromone. We propose that sensillar pheromone-degrading enzymes modulate pheromone concentration in the receptor space and hence play a dynamic role in the pheromone-mediated reproductive behaviors of these animals. PMID:2246254

  15. Functional morphology of the ommatidia in the compound eye of the moth, Antheraea polyphemus (Insecta, Saturniidae).

    PubMed

    Anton-Erxleben, F; Langer, H

    1988-05-01

    The fine structure of the superposition eye of the Saturniid moth Antheraea polyphemus Cramer was investigated by electron microscopy. Each of the approximately 10,000 ommatidia consists of the same structural components, but regarding the arrangement of the ommatidia and the rhabdom structure therein, two regions of the eye have to be distinguished. In a small dorsal rim area, the ommatidia are characterized by rectangularly shaped rhabdoms containing parallel microvilli arranged in groups that are oriented perpendicular to each other. In all other ommatidia, the proximal parts of the rhabdoms show radially arranged microvilli, whereas the distal parts may reveal different patterns, frequently with microvilli in two directions or sometimes even in one direction. Moreover, the microvilli of all distal cells are arranged in parallel to meridians of the eyes. By virtue of these structural features the eyes should enable this moth not only discrimination of the plane of polarized light but also skylight-orientation via the polarization pattern, depending on moon position. The receptor cells exhibit only small alterations during daylight within the natural diurnal cycle. However, under illumination with different monochromatic lights of physiological intensity, receptor cells can be unbalanced: Changes in ultrastructure of the rhabdomeres and the cytoplasm of such cells are evident. The effects are different in the daytime and at night. These findings are discussed in relation to the breakdown and regeneration of microvilli and the influence of the diurnal cycle. They are compared with results on photoreceptor membrane turnover in eyes of other arthropod species. PMID:3383218

  16. 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. PMID:17173967

  17. Molecular cloning and characterization of Antheraea mylitta cytoplasmic polyhedrosis virus polyhedrin gene and its variant forms

    SciTech Connect

    Sinha-Datta, Uma [Department of Biotechnology, Indian Institute of Technology, Kharagpur 721302 (India); Chavali, Venkata Ramana Murthy [Department of Biotechnology, Indian Institute of Technology, Kharagpur 721302 (India); Ghosh, Ananta K. [Department of Biotechnology, Indian Institute of Technology, Kharagpur 721302 (India)]. E-mail: aghosh@hijli.iitkgp.ernet.in

    2005-07-08

    The segments 10 (S10) of the 11 double stranded RNA genomes from Antheraea mylitta cytoplasmic polyhedrosis virus (AmCPV) encoding a novel polyhedrin polypeptide was converted to cDNA, cloned, and sequenced. Three cDNA clones consisting of 1502 (AmCPV10-1), 1120 (AmCPV10-2), and 1415 (AmCPV10-3) nucleotides encoding polyhedrin of 254, 339, and 319 amino acids with molecular masses of 29, 39, and 37 kDa, respectively, were obtained, and verified by Northern analysis. These clones showed 70-94% sequence identity among them but none with any sequences in databases. The expression of AmCPV10-1 cDNA encoded polyhedrin in Sf-9 cells was detected by immunoblot analysis and formation of polyhedra by electron microscopy, as observed in AmCPV-infected gut cells, but no expression of AmCPV10-2 or AmCPV10-3 cDNA was detected, indicating that during AmCPV replication, along with functional S10 RNA, some defective variant forms of S10 RNAs are packaged in virion particles.

  18. Molecular cloning and expression analysis of a hemolin-like molecule from Antheraea pernyi.

    PubMed

    Sun, Yu; Wang, Lei; Qian, Cen; Dai, Lishang; Li, Jun; Zhang, Congfen; Zhu, Bao-Jian; Liu, Chao-Liang

    2015-05-01

    Hemolin belongs to the immunoglobulin superfamily and plays an important role in innate immune response of insects. In this study, a hemolin-like cDNA of 1418bp was obtained from Antheraea pernyi (Ap-hemolin-like). Sequence analysis revealed Ap-hemolin-like was homologous to those hemolins from other insect species. Recombinant Ap-hemolin-like protein was expressed in Escherichia coli cells, and polyclonal antibodies were produced against the recombinant proteins. Real-time PCR and western blot analysis showed that the Ap-hemolin-like was expressed in hemolymph, Malpighian tubules, midgut, epidermis and fat body, with the highest expression level in hemolymph. To investigate its role in the immune response against microorganisms, fifth instar larvae were challenged by injecting nucleopolyhedrovirus (NPV), E. coli, or Beauveria bassiana. The results showed that the expression of Ap-hemolin-like in hemolymph and fat body was obviously induced by microorganisms. In addition, the recombinant Ap-hemolin-like protein promoted the agglutination of E. coli in the presence of calcium, which was confirmed by agglutination assay. These results suggested that the Ap-hemolin-like protein was involved in innate immune response of A. pernyi against pathogens. PMID:25812765

  19. 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. PMID:24867887

  20. 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.

  1. A receptor and binding protein interplay in the detection of a distinct pheromone component in the silkmoth Antheraea polyphemus.

    PubMed

    Forstner, Maike; Breer, Heinz; Krieger, Jürgen

    2009-01-01

    Male moths respond to conspecific female-released pheromones with remarkable sensitivity and specificity, due to highly specialized chemosensory neurons in their antennae. In Antheraea silkmoths, three types of sensory neurons have been described, each responsive to one of three pheromone components. Since also three different pheromone binding proteins (PBPs) have been identified, the antenna of Antheraea seems to provide a unique model system for detailed analyzes of the interplay between the various elements underlying pheromone reception. Efforts to identify pheromone receptors of Antheraea polyphemus have led to the identification of a candidate pheromone receptor (ApolOR1). This receptor was found predominantly expressed in male antennae, specifically in neurons located beneath pheromone-sensitive sensilla trichodea. The ApolOR1-expressing cells were found to be surrounded by supporting cells co-expressing all three ApolPBPs. The response spectrum of ApolOR1 was assessed by means of calcium imaging using HEK293-cells stably expressing the receptor. It was found that at nanomolar concentrations ApolOR1-cells responded to all three pheromones when the compounds were solubilized by DMSO and also when DMSO was substituted by one of the three PBPs. However, at picomolar concentrations, cells responded only in the presence of the subtype ApolPBP2 and the pheromone (E,Z)-6,11-hexadecadienal. These results are indicative of a specific interplay of a distinct pheromone component with an appropriate binding protein and its related receptor subtype, which may be considered as basis for the remarkable sensitivity and specificity of the pheromone detection system. PMID:20011135

  2. 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.

  3. Specificity Determinants of the Silkworm Moth Sex Pheromone

    PubMed Central

    Xu, Pingxi; Hooper, Antony M.; Pickett, John A.; Leal, Walter S.

    2012-01-01

    The insect olfactory system, particularly the peripheral sensory system for sex pheromone reception in male moths, is highly selective, but specificity determinants at the receptor level are hitherto unknown. Using the Xenopus oocyte recording system, we conducted a thorough structure-activity relationship study with the sex pheromone receptor of the silkworm moth, Bombyx mori, BmorOR1. When co-expressed with the obligatory odorant receptor co-receptor (BmorOrco), BmorOR1 responded in a dose-dependent fashion to both bombykol and its related aldehyde, bombykal, but the threshold of the latter was about one order of magnitude higher. Solubilizing these ligands with a pheromone-binding protein (BmorPBP1) did not enhance selectivity. By contrast, both ligands were trapped by BmorPBP1 leading to dramatically reduced responses. The silkworm moth pheromone receptor was highly selective towards the stereochemistry of the conjugated diene, with robust response to the natural (10E,12Z)-isomer and very little or no response to the other three isomers. Shifting the conjugated diene towards the functional group or elongating the carbon chain rendered these molecules completely inactive. In contrast, an analogue shortened by two omega carbons elicited the same or slightly higher responses than bombykol. Flexibility of the saturated C1–C9 moiety is important for function as addition of a double or triple bond in position 4 led to reduced responses. The ligand is hypothesized to be accommodated by a large hydrophobic cavity within the helical bundle of transmembrane domains. PMID:22957053

  4. Evaluation of general toxicity and genotoxicity of the silkworm extract powder.

    PubMed

    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; Kang, Boo-Hyon

    2013-12-31

    The silkworm extract powder contain 1-deoxynojirimycin (DNJ), a potent ?-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

  5. mRNA populations during wing development in the silkmoth Antheraea polyphemus.

    PubMed

    Katula, K; Gilbert, L I; Sridhara, S

    1981-12-01

    Pupal wing tissue of the American silkmoth Antheraea polyphemus has been used as a model system to study 20-hydroxyecdysone and juvenile hormone control of cuticle protein synthesis. Juvenile hormone does not affect either the content or rate of synthesis of RNA and protein of the wing tissue. both of which show linear increases during the first few days of hormonal treatment. Based on the fractionation of total RNA on oligo-dT columns the percent of mRNA remains the same throughout development after both hormone treatments. However, both the amount of poly-A+ RNA in the wing tissue, and its content of poly-A show considerable increases as a function of development. The products of translation of the various poly-A+ RNA populations in the cell-free wheatgerm system have been analyzed by one- and two-dimensional gel electrophoresis and fluorography. Qualitative changes occur during the first 24 h; the production of a mRNA coding for a protein of approx. 40 000 dalton is stimulated and the production of a mRNA coding for a protein of 29 000 dalton is greatly reduced. Only a few differences are observed between samples from the 2 hormone treatments. Over the next 5-15 days of development mainly quantitative changes are observed. Juvenile hormone application results in quantitative changes in specific mRNAs, but no new mRNAs unique to juvenile hormone action are observed. The data are consistent with the concept that in altering the epidermal developmental program, juvenile hormone is apparently modulating the action of 20-hydroxyecdysone. PMID:6173272

  6. 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

  7. Snmp1, a Novel Membrane Protein of Olfactory Neurons of the Silk Moth Antheraea polyphemus with Homology to the CD36 Family of Membrane Proteins

    Microsoft Academic Search

    Matthew E. Rogers; Ming Sun; Michael R. Lerner; Richard G. Vogt

    1997-01-01

    While olfactory neurons of silk moths are well known for their exquisite sensitivity to sex pheromone odor- ants, molecular mechanisms underlying this sensitivity are poorly understood. In searching for proteins that might support olfactory mechanisms, we characterized the protein profile of olfactory neuron receptor mem- branes of the wild silk moth Antheraea polyphemus .W e have purified and cloned a

  8. Species-specific pheromonal compounds induce distinct conformational changes of pheromone binding protein subtypes from Antheraea polyphemus.

    PubMed

    Mohl, Claudia; Breer, Heinz; Krieger, Jürgen

    2002-10-01

    We have investigated the structural features of three pheromone binding protein (PBP) subtypes from Antheraea polyphemus and monitored possible changes induced upon interaction with the Antheraea pheromonal compounds 4E,9Z-14:Ac [(E4,Z9)-tetradecadienyl-1-acetate], 6E,11Z-16:Ac [(E6,Z11)-hexadecadienyl-1-acetate], and 6E,11Z-16:Al [(E6,Z11)-hexadecadienal]. Circular dichroism and second derivative UV-difference spectroscopy data demonstrate that the structure of subtype PBP1 significantly changes upon binding of 4E,9Z-14:Ac. The related 6E,11Z-16:Ac was less effective and 6E,11Z-16:Al showed only a small effect. In contrast, in subtype PBP2 pronounced structural changes were only induced by the 6E,11Z-16:Al, and the subtype PBP3 did not show any considerable changes in response to the pheromonal compounds. The UV-spectroscopic data suggest that histidine residues are likely to be involved in the ligand-induced structural changes of the proteins, and this notion was confirmed by site-directed mutagenesis experiments. These results demonstrate that appropriate ligands induce structural changes in PBPs and provide evidence for ligand specificity of these proteins. PMID:12488967

  9. Analysis of Genetic Divergence for Classification of Morphological and Larval Gain Characteristics of Peanut Cocoon Silkworm (Bombyx mori L.) Germplasm

    Microsoft Academic Search

    M. Salehi Nezhad; S. Z. Mirhosseini

    2009-01-01

    A hierarchical agglomerative clustering analysis was undertaken for grouping the 51 lines silkworm, Bombyx mori L., based on larval gains parameters in the clustering process. The analysis was based on data from one rearing seasons with all 51 peanut cocoon strains of silkworm and varying morphological development potentials. The results indicate that two clusters can be realized based on larval

  10. New and highly efficient expression systems for expressing selectively foreign protein in the silk glands of transgenic silkworm.

    PubMed

    Zhao, Aichun; Zhao, Tianfu; Zhang, Yuansong; Xia, Qingyou; Lu, Cheng; Zhou, Zeyang; Xiang, Zhonghuai; Nakagaki, M

    2010-02-01

    We constructed three different fibroin H-chain expression systems to estimate the efficacy of producing recombinant proteins in the cocoon of transgenic silkworms. The results showed that the three different EGFP/H-chain fusion genes were all expressed selectively in the posterior silk gland of the transgenic silkworm. The recombinant protein content of transgenic silkworm cocoons is up to 15% (w/w) when using the most highly efficient H-chain expression system. To our knowledge, in comparison with silkworm silk gland expression systems in the literature, the highly efficient expression system developed in this study is the most efficient silkworm silk gland expression system to date. This expression system is the best candidate for foreign gene production and for creation of novel functional silk material. The results suggested the N-terminal domain and the intron of the H-chain gene are important in the secretion of fibroin and its transcription, respectively. PMID:19533404

  11. 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

  12. Expression, purification and antigenicity of Neospora caninum-antigens using silkworm larvae targeting for subunit vaccines.

    PubMed

    Otsuki, Takahiro; Dong, Jinhua; Kato, Tatsuya; Park, Enoch Y

    2013-02-18

    Infection of Neospora caninum causes abortion in cattle, which has a serious worldwide impact on the economic performance of the dairy and beef industries. Now, inexpensive and efficacious vaccines are required to protect cattle from neosporosis in livestock industry. In this study, N. caninum surface antigen 1 (SAG1) and SAG1-related sequence 2 (SRS2) were expressed in hemolymph of silkworm larvae as a soluble form. Expressed SAG1 and SRS2 clearly showed antigenicity against N. caninum-positive sera of cow. SAG1 and SRS2 were purified to near homogeneity from hemolymph of silkworm larvae using anti-FLAG M2 antibody agarose: approximately 1.7 mg of SAG1 from 10 silkworm larvae and 370 ?g of SRS2 from 17 silkworm larvae. Mice that were injected by antigens induced antibodies against SAG1 and SRS2. This study indicates that it is possible that this silkworm expression system leads to a large-scale production of N. caninum-antigens with biological function and low production cost. Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid expression system paves the way to produce largely and rapidly these recombinant antigens for its application to subunit vaccines against neosporosis in cattle. PMID:23102762

  13. 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. PMID:26055521

  14. Space flight experiment on chinese silkworm on board the Russian 10th biosatellite

    NASA Astrophysics Data System (ADS)

    Zhizhen, Shi; Dahuan, Zhuang; Ilyin, Eugene A.

    Space flight experiments on Chinese silkworm (Bombyx mori L.) were conducted on board the Russian 10th Biosatellite for 12 days. The samples included silkworm eggs, larvae, cocoons, pupae and moths. The processes of spinning, cocooning, mating, oviposition, larval hatching, pupation and moth emergence all completed well in space. The following effects of space flight on silkworm development were observed: The times of hatching and oviposition in the flight group were 2 to 3 days earlier than in the control group; the hatching rate of diapause eggs during space flight seemed higher than that of the control group; the life span of 2 of the 7 varieties flown was shortened; genetical variations appeared in 3 varieties. The results showed that the embryonic stage was probably the period most sensitive to the space flight environment.

  15. Assessment of genetic diversity by DNA profiling and its significance in silkworm, Bombyx mori.

    PubMed

    Nagaraju, J G; Singh, L

    1997-08-01

    Silkworm genetic resources that are being maintained in different countries are yet to be adequately tapped to develop elite varieties that are suited to different agro-eco-climatic conditions of countries like India. This is mostly due to unavailability of efficient protocols that could uncover usable genetic variability in silkworms. Molecular markers are known to provide unambiguous estimates of genetic variability of populations since they are independent of confounding effects of environment. The DNA fingerprinting assays, based on random amplified polymorphic DNA (RAPD) and banded krait minor satellite DNA (Bkm) 2(8) multilocus probes, which successfully characterise the diverse silkworm genotypes at their DNA level, are described. The use of these two DNA fingerprinting assays in estimation of within- and between-population genetic diversity is discussed. PMID:9378144

  16. A novel granulocyte-specific ? 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 ? subunit of integrin, Bmintegrin?PS3, as being specifically and highly expressed in silkworm haemocytes. Immunofluorescence analysis validated the specificity of Bmintegrin?PS3 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 Bmintegrin?PS3 could be used as a specific marker for granulocytes and could be applied to future molecular cell biology studies. PMID:25450560

  17. 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.

  18. Vitellogenin from the Silkworm, Bombyx mori: An Effective Anti-Bacterial Agent

    PubMed Central

    Kumar, Manish; Prasad, Tulika; Kannan, Mani; König, Simone

    2013-01-01

    Silkworm, Bombyx mori, vitellogenin (Vg) was isolated from perivisceral fat body of day 3 of pupa. Both Vg subunits were co-purified as verified by mass spectrometry and immunoblot. Purified Vg responded to specific tests for major posttranslational modifications on native gels indicating its nature as lipo-glyco-phosphoprotein. The Vg fraction had strong antibacterial activity against Gram negative bacterium Escherichia coli and Gram positive bacterium Bacillus subtilis. Microscopic images showed binding of Vg to bacterial cells and their destruction. When infected silkworm larvae were treated with purified Vg they survived the full life cycle in contrast to untreated animals. This result showed that Vg has the ability to inhibit the proliferation of bacteria in the silkworm fluid system without disturbing the regular metabolism of the host. PMID:24058454

  19. 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. PMID:25113390

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

    PubMed Central

    2009-01-01

    Background Carboxylesterase is a multifunctional superfamily and ubiquitous in all living organisms, including animals, plants, insects, and microbes. It plays important roles in xenobiotic detoxification, and pheromone degradation, neurogenesis and regulating development. Previous studies mainly used Dipteran Drosophila and mosquitoes as model organisms to investigate the roles of the insect COEs in insecticide resistance. However, genome-wide characterization of COEs in phytophagous insects and comparative analysis remain to be performed. Results Based on the newly assembled genome sequence, 76 putative COEs were identified in Bombyx mori. Relative to other Dipteran and Hymenopteran insects, alpha-esterases were significantly expanded in the silkworm. Genomics analysis suggested that BmCOEs showed chromosome preferable distribution and 55% of which were tandem arranged. Sixty-one BmCOEs were transcribed based on cDNA/ESTs and microarray data. Generally, most of the COEs showed tissue specific expressions and expression level between male and female did not display obvious differences. Three main patterns could be classified, i.e. midgut-, head and integument-, and silk gland-specific expressions. Midgut is the first barrier of xenobiotics peroral toxicity, in which COEs may be involved in eliminating secondary metabolites of mulberry leaves and contaminants of insecticides in diet. For head and integument-class, most of the members were homologous to odorant-degrading enzyme (ODE) and antennal esterase. RT-PCR verified that the ODE-like esterases were also highly expressed in larvae antenna and maxilla, and thus they may play important roles in degradation of plant volatiles or other xenobiotics. Conclusion B. mori has the largest number of insect COE genes characterized to date. Comparative genomic analysis suggested that the gene expansion mainly occurred in silkworm alpha-esterases. Expression evidence indicated that the expanded genes were specifically expressed in midgut, integument and head, implying that these genes may have important roles in detoxifying secondary metabolites of mulberry leaves, contaminants in diet, and odorants. Our results provide some new insights into functions and evolutionary characteristics of COEs in phytophagous insects. PMID:19930670

  1. Silkworms can be used as an animal model to screen and evaluate gouty therapeutic drugs.

    PubMed

    Zhang, Xiaoli; Xue, Renyu; Cao, Guangli; Pan, Zhonghua; Zheng, Xiaojian; Gong, Chengliang

    2012-01-01

    In the past few decades, the mouse has been used as a mammalian model for hyperuricemia and gout, which has increased not only in prevalence, but also in clinical complexity, accentuated in part by a dearth of novel advances in treatments for hyperuricemia and gouty arthritis. However, the use of mice for the development of gouty therapeutic drugs creates a number of problems. Thus, identification and evaluation of the therapeutic effects of chemicals in an alternative animal model is desirable. In the present study, the effects of gouty therapeutic drugs on lowering the content of uric acid and inhibiting activity of xanthine oxidase were evaluated by using a silkworm model, Bombyx mori L. (Lepidoptera: Bombycidae). The results showed that the effectiveness of oral administration of various gouty therapeutic drugs to 5(th) instar silkworms is consistent with results for human. The activity of xanthine oxidase of silkworm treated with allopurinol was lower, and declined in a dose-dependent manner compared with control silkworms, while sodium bicarbonate failed at inhibiting the activity of xanthine oxidase. The concentration of uric acid in the both hemolymph and fat body declined by 90 and 95% at six days post-administration with 25 mg/mL of allopurinol, respectively (p < 0.01), while the concentration of uric acid in both the hemolymph and fat body also declined by 81 and 95% at six days post-administration with 25 mg/mL of sodium bicarbonate, respectively (p < 0.01). Moreover, the epidermis of silkworm treated with allopurinol or sodium bicarbonate became transparent compared with the negative control group. These results suggest that silkworm larva can be used as an animal model for screening and evaluation of gouty therapeutic drugs. PMID:22934965

  2. Evidence of selection at melanin synthesis pathway loci during silkworm domestication.

    PubMed

    Yu, Hong-Song; Shen, Yi-Hong; Yuan, Gang-Xiang; Hu, Yong-Gang; Xu, Hong-En; Xiang, Zhong-Huai; Zhang, Ze

    2011-06-01

    The domesticated silkworm (Bombyx mori) was domesticated from wild silkworm (Bombyx mandarina) more than 5,000 years ago. During domestication, body color between B. mandarina and B. mori changed dramatically. However, the molecular mechanism of the silkworm body color transition is not known. In the present study, we examined within- and between-species nucleotide diversity for eight silkworm melanin synthesis pathway genes, which play a key role in cuticular pigmentation of insects. Our results showed that the genetic diversity of B. mori was significantly lower than that of B. mandarina and 40.7% of the genetic diversity of wild silkworm was lost in domesticated silkworm. We also examined whether position effect exists among melanin synthesis pathway genes in B. mandarina and B. mori. We found that the upstream genes have significantly lower levels of genetic diversity than the downstream genes, supporting a functional constraint hypothesis (FCH) of metabolic pathway, that is, upstream enzymes are under greater selective constraint than downstream enzymes because upstream enzymes participate in biosynthesis of a number of metabolites. We also investigated whether some of the melanin synthesis pathway genes experienced selection during domestication. Neutrality test, coalescent simulation, as well as network and phylogenetic analyses showed that tyrosine hydroxylase (TH) gene was a domestication locus. Sequence analysis further suggested that a putative expression enhancer (Abd-B-binding site) in the intron of TH gene might be disrupted during domestication. TH is the rate-limiting enzyme of melanin synthesis pathway in insects. Real-time polymerase chain reaction assay did show that the relative expression levels of TH gene in B. mori were significantly lower than that in B. mandarina at three different developmental stages, which is consistent with light body color of domesticated silkworm relative to wild silkworm. Therefore, we speculated that expression change of TH gene may contribute to the body color transition from B. mandarina to B. mori. Our results emphasize the exceptional role of gene expression regulation in morphological transition of domesticated animals. PMID:21212153

  3. Nature of heterosis and combining ability in the silkworm.

    PubMed

    Strunnikov, V A

    1986-07-01

    The isogenic, highly heterotic parthenoclone 29, originating from a hybrid silkworm female, was transformed via unisexual reproduction (meiotic and ameiotic parthenogenesis) into four genotypical variants differing in well-known various levels of hetero zygosity and combinations of useful and harmful genes. A comparison of these changes with the heterosis level made it possible to discover that both heterosity for adaptively neutral genes (overdominance hypothesis) and the number of allelic pairs, each of them being heterozygous for a favourable, completely dominant gene (dominance hypothesis) play no decisive role in the intensity of heterosis. The level of heterosis is largely determined by the relationship between the effects of useful and harmful genes, the first falling into the category of semidominant, cumulatively acting genes which control viability. Their favourable, joint well-coordinated effects, unlike those of genes which control quantitative characters, increase in relation to the number of genes in a geometric rather than an arithmetic progression. The interaction between semilethal genes is subjected to the same regularity. The high combining ability of parthenoclone 29 variants is determined by the number and homozygosity of the useful genes. PMID:24248023

  4. Measured behavioural latency in response to sex-pheromone loss in the large silk moth Antheraea polyphemus.

    PubMed

    Baker, T C; Vogt, R G

    1988-07-01

    Males of the giant silk moth Antheraea polyphemus Cramer (Lepidoptera: Saturniidae) were video-recorded in a sustained-flight wind tunnel in a constant plume of sex pheromone. The plume was experimentally truncated, and the moths, on losing pheromone stimulus, rapidly changed their behaviour from up-tunnel zig-zag flight to lateral casting flight. The latency of this change was in the range 300-500 ms. Video and computer analysis of flight tracks indicates that these moths effect this switch by increasing their course angle to the wind while decreasing their air speed. Combined with previous physiological and biochemical data concerning pheromone processing within this species, this behavioural study supports the argument that the temporal limit for this behavioural response latency is determined at the level of genetically coded kinetic processes located within the peripheral sensory hairs. PMID:3209970

  5. Co-Sensitization to Silkworm Moth (Bombyx mori) and 9 Inhalant Allergens among Allergic Patients in Guangzhou, Southern China

    PubMed Central

    Wei, Nili; Huang, Huimin; Zeng, Guangqiao

    2014-01-01

    Objectives This study aimed to investigate the profile of sensitization to silkworm moth (Bombyx mori) and other 9 common inhalant allergens among patients with allergic diseases in southern China. Methods A total of 175 patients were tested for serum sIgE against silkworm moth in addition to combinations of other allergens: Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicalis, Blattella germanica, Periplaneta americana, cat dander, dog dander, Aspergillus fumigatus and Artemisia vulgaris by using the ImmunoCAP system. Correlation between sensitization to silkworm moth and to the other allergens was analyzed. Results Of the 175 serum samples tested, 86 (49.14%) were positive for silkworm moth sIgE. With high concordance rates, these silkworm moth sensitized patients were concomitantly sensitized to Dermatophagoides pteronyssinus (94.34%), Dermatophagoides farinae (86.57%), Blomia tropicalis (93.33%), Blattella germanica (96.08%), and Periplaneta americana (79.41%). Moreover, there was a correlation in serum sIgE level between silkworm moth and Dermatophagoides pteronyssinus (r?=?0.518), Dermatophagoides farinae (r?=?0.702), Blomia tropicalis (r?=?0.701), Blattella germanica (r?=?0.878), and Periplaneta americana (r?=?0.531) among patients co-sensitized to silkworm moth and each of these five allergens. Conclusion In southern Chinese patients with allergic diseases, we showed a high prevalence of sensitization to silkworm moth, and a co-sensitization between silkworm moth and other five common inhalant allergens. Further serum inhibition studies are warranted to verify whether cross-reactivity exists among these allergens. PMID:24787549

  6. Cyclic GMP levels and guanylate cyclase activity in pheromone-sensitive antennae of the silkmoths Antheraea polyphemus and Bombyx mori.

    PubMed

    Ziegelberger, G; van den Berg, M J; Kaissling, K E; Klumpp, S; Schultz, J E

    1990-04-01

    Female sex pheromones applied to freshly isolated, living antennae of male Antheraea polyphemus and Bombyx mori led to an increase of cGMP. A 1:1 mixture of 2 pheromone components of Antheraea polyphemus blown for 10 sec in physiological concentrations over their antennal branches raised cGMP levels about 1.34-fold (+/- 0.08 SEM, n = 23) from a basal level of 3.0 +/- 0.6 (SEM, n = 20) pmol/mg protein. Similarly, bombykol elicited a 1.29-fold (+/- 0.13 SEM, n = 23) cGMP increase in antennae of male Bombyx mori from a basal level of 2.7 +/- 0.5 (SEM, n = 24) pmol/mg protein. No cross-sensitivity was found with respect to pheromones from either species. In antennae of female silkmoths, the cGMP response was missing upon stimulation with their own respective pheromones according to the known lack of pheromone receptor cells in the female. cAMP levels in the male antennae of 14.2 +/- 2.9 (SEM, n = 4) pmol/mg protein in A. polyphemus and 15.0 +/- 3.0 (SEM, n = 5) pmol/mg protein in B. mori were not affected by pheromone stimulation. Within 1-60 sec, the extent of cGMP increase in B. mori was independent of the duration of pheromone exposure. The levels of cGMP in pheromone-stimulated antennae of both species remained elevated for at least 10 min, i.e., much longer than the duration of the receptor potential measured in single-cell recordings. Guanylate cyclase activity was identified in homogenates of male and female antennae from both species. The Km of the guanylate cyclase from male B. mori for the preferential substrate MnGTP was 175 microM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1970356

  7. Study of Protein Conformation and Orientation in Silkworm and Spider Silk Fibers Using Raman Microspectroscopy

    Microsoft Academic Search

    Marie-Eve Rousseau; Thierry Lefèvre; Lilyane Beaulieu; Tetsuo Asakura; Michel Pézolet

    2004-01-01

    Raman microspectroscopy has been used for the first time to determine quantitatively the orientation of the ‚-sheets in silk monofilaments from Bombyx moriand Samia cynthia ricini silkworms, and from the spider Nephila edulis. It is shown that, for systems with uniaxial symmetry such as silk, it is possible to determine the order parameters ?P2? and ?P4? of the orientation distribution

  8. 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

  9. 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

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

    Microsoft Academic Search

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

    2008-01-01

    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

  11. Biosynthesis and cocoon-export of a recombinant globular protein in transgenic silkworms

    Microsoft Academic Search

    Corinne Royer; Audrey Jalabert; Martine Da Rocha; Anne-Marie Grenier; Bernard Mauchamp; Pierre Couble; Gérard Chavancy

    2005-01-01

    A gene construct was made by fusing the coding sequence of the red fluorescent protein (DsRed) to the exon 2 of the fibrohexamerin gene (fhx), that encodes a subunit of fibroin, the major silk protein of the silkworm Bombyx mori. The fusion gene was inserted into a piggyBac vector to establish a series of transgenic lines. The expression of the

  12. A potential material for tissue engineering: Silkworm silk\\/PLA biocomposite

    Microsoft Academic Search

    Hoi-Yan Cheung; Kin-Tak Lau; Xiao-Ming Tao; David Hui

    2008-01-01

    Poly(lactic acid) (PLA), a kind of well recognized biodegradable polymer, was reinforced by silkworm silk fibers to form a completely biodegradable and biocompatible biocomposite for tissue engineering applications. The influence on the mechanical and thermal properties of the biocomposite in relation to the length and weight content of silk fibers is studied in this paper. Through the micro-hardness test, optimized

  13. Cytoplasmic polyhedrosis virus-induced differential gene expression in two silkworm strains of different susceptibility.

    PubMed

    Gao, Kun; Deng, Xiang-Yuan; Qian, He-Ying; Qin, Guang-Xing; Hou, Cheng-Xiang; Guo, Xi-Jie

    2014-04-15

    Digital gene expression (DGE) was performed to investigate the gene expression profiles of 4008 and p50 silkworm strains at 48 h after oral infection with BmCPV. 3,668,437 clean tags were identified in the BmCPV-infected p50 silkworms and 3,540,790 clean tags in the control p50. By contrast, 4,498,263 clean tags were identified in the BmCPV-infected 4008 silkworms and 4,164,250 clean tags in the control 4008. A total of 691 differentially expressed genes were detected in the infected 4008 DGE library and 185 were detected in the infected p50 DGE library, respectively. The expression profiles identified some important differentially expressed genes involved in signal transduction, enzyme activity and apoptotic changes, some of which were verified using quantitative real-time PCR (qRT-PCR). These results provide important clues on the molecular mechanism of BmCPV invasion and resistance mechanism of silkworms against BmCPV infection. PMID:24525400

  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. PMID:25162624

  15. Structural Basis of Ligand Binding and Release in Insect Pheromone-binding Proteins: NMR Structure of Antheraea polyphemus PBP1 at pH 4.5

    Microsoft Academic Search

    Fred F. Damberger; Yuko Ishida; Walter S. Leal; Kurt Wüthrich

    2007-01-01

    The NMR structure of the Antheraea polyphemus pheromone-binding protein 1 at pH 4.5, ApolPBP1A, was determined at 20 °C. The structure consists of six ?-helices, which are arranged in a globular fold that encapsulates a central helix ?7 formed by the C-terminal polypeptide segment 131–142. The 3D arrangement of these helices is anchored by the three disulfide bonds 19–54, 50–108 and

  16. Immunolocalization of pheromone-binding protein and general odorant-binding protein in olfactory sensilla of the silk moths Antheraea and Bombyx

    Microsoft Academic Search

    R. A. Steinbrecht; M. Laue; G. Ziegelberger

    1995-01-01

    The distribution of odorant-binding proteins among olfactory sensilla of three moth species was studied by immuno-electron microscopy. Two polyclonal antisera were used in a post-embedding labelling protocol on sections of cryo-substituted antennae. The first was directed against the pheromone-binding protein (PBP) of Antheraea polyphemus, the second against the general odorant-binding protein (GOBP) of the same species. Immunoblots showed that these

  17. SUDDEN OAK DEATH Field Meeting

    E-print Network

    California at Berkeley, University of

    SUDDEN OAK DEATH Field Meeting Saturday, December 3, 1:00 p.m. Joaquin Miller Park - Craib Picnic Area (if raining, come to the Ranger Station) Learn about Sudden Oak Death management and see Death (SOD). SOD is a fungus-like mold that is killing oak trees in coastal California. Several samples

  18. Expression profile analysis of silkworm P450 family genes after phoxim induction.

    PubMed

    Li, Fanchi; Ni, Min; Zhang, Hua; Wang, Binbin; Xu, Kaizun; Tian, Jianghai; Hu, Jingsheng; Shen, Weide; Li, Bing

    2015-07-01

    Silkworm (Bombyx mori) is an important economic insect and a model species for Lepidopteran. Each year, O,O-diethyl O-(alpha-cyanobenzylideneamino) phosphorothioate (phoxim) pesticide poisoning in China results in huge economic losses in sericulture. Silkworm fat body is the main organ for nutrient storage, energy supply, intermediary metabolism, and detoxification. Microarray analysis of silkworm Cytochrome P450 detoxification enzyme genes revealed that all tested P450 4 (CYP4) family genes are expressed in the fat body. Quantitative Real-time PCR (QRT-PCR) was used to detect the expression of CYP4 family genes in silkworm fat body 0, 24, 48, and 72?h after phoxim exposure. The expression levels of silkworm molting hormone synthesis-related genes started to change 24?h after phoxim exposure, with those of CYP302A1, CYP306A1, and CYP314A1 being elevated by 1.38-, 1.33-, and 2.10-fold, respectively. The CYP18A1 gene that participates in steroid hormone inactivation and the CYP15C1 gene that participates in the epoxidation during the synthesis of juvenile hormone (JH) from methyl farnesoate (MF) were increased by 3.85- and 7.82-fold, respectively. Phylogenetic analysis indicated that these endogenous hormone metabolism-related genes belong to CYP mito clan and clan 2, and that phoxim exposure may affect silkworm development and metamorphosis. The CYP4, CYP6, and CYP9 families all showed some degrees of increases in gene expression; among them, CYP49A1, CYP4L6, CYP6AB4, CYP9G3, CYP9A19, and CYP9A22's transcription levels were significantly upregulated to 12.77-, 2.64-, 2.42-, 4.06-, 3.32-, and 2.98-fold, respectively, of the control levels. In the fat body, CYP49A1, CYP6AB4, CYP9A19, and CYP9A22 were constantly expressed at high levels after 24, 48, and 72?h of phoxim treatments; according to phylogenetic analysis, these genes belong to detoxification-related clan 3 and clan 4 CYP families. These genes may participate in the metabolism of phoxim in silkworm fat body. The results obtained in this study provide a basis for future in-depth investigations of insect P450 family genes in metabolic detoxification. PMID:26071814

  19. Identification of a novel strong and ubiquitous promoter/enhancer in the silkworm Bombyx mori.

    PubMed

    Tsubota, Takuya; Uchino, Keiro; Suzuki, Takao K; Tanaka, Hiromitsu; Kayukawa, Takumi; Shinoda, Tetsuro; Sezutsu, Hideki

    2014-07-01

    Transgenic techniques offer a valuable tool for determining gene functions. Although various promoters are available for use in gene overexpression, gene knockdown, and identification of transgenic individuals, there is nevertheless a lack of versatile promoters for such studies, and this dearth acts as a bottleneck, especially with regard to nonmodel organisms. Here, we succeeded in identifying a novel strong and ubiquitous promoter/enhancer in the silkworm. We identified a unique silkworm strain whose reporter gene showed strong and ubiquitous expression during the establishment of enhancer trap strains. In this strain, the transposon was inserted into the 5'UTR of hsp90, a housekeeping gene that is abundantly expressed in a range of tissues. To determine whether the promoter/enhancer of hsp90 could be used to induce strong gene expression, a 2.9-kb upstream genomic fragment of hsp90 was isolated (hsp90(P2.9k)), and its transcriptional activation activity was examined. Strikingly, hsp90(P2.9k) induced strong gene expression in silkworm cell cultures and also strongly induced gene expression in various tissues and developmental stages of the silkworm. hsp90(P2.9k) also exhibited significant promoter/enhancer activity in Sf9, a cell culture from the armyworm, suggesting that this fragment might possibly be used as a gene expression tool in other Lepidoptera. We further found that 2.0 kb of hsp90(P2.9k) is sufficient for the induction of strong gene expression. We believe that this element will be of value for a range of studies such as targeted gene overexpression, gene knockdown and marker gene expression, not only in the silkworm but also in other insect species. PMID:24875626

  20. Cloning and expression analysis of a peptidoglycan recognition protein in silkworm related to virus infection.

    PubMed

    Gao, Kun; Deng, Xiang-Yuan; Qian, He-Ying; Qin, Guang-Xing; Hou, Cheng-Xiang; Guo, Xi-Jie

    2014-11-15

    In this study, the full-length cDNA of a peptidoglycan recognition protein named BmPGRP-S3 was identified from the silkworm, Bombyx mori by rapid amplification of cDNA ends. It is 807 bp and comprises the following: a 5'-untranslated region (UTR) with a length of 112 bp, a 3'-UTR with a length of 92 bp including a poly-adenylation signal sequence (AATAAA) and a poly(A) tail. The longest open reading frame (ORF) of BmPGRP-S3 is 603 bp and encodes a polypeptide of 200 amino acids with a predicted molecular weight of 22.3 kDa including a PGRP domain. Sequence similarity and phylogenic analysis results indicated that BmPGRP-S3 belongs to the group of insect PGRPs and is closer to BmPGRP-S4 with the highest identity of 68%. Fluorescent quantitative real-time PCR results revealed that the mRNA transcripts of BmPGRP-S3 were presented in all of the tissues, but were highest in the midgut. In the silkworm larvae infected with B. mori cytoplasmic polyhedrosis virus (BmCPV), the relative expression level of BmPGRP-S3 was upregulated. The DNA segment of a mature BmPGRP-S3 peptide was inserted into the expression plasmid pET-28a(+) to construct a recombinant expression plasmid. Western blot results revealed that mature BmPGRP-S3 could be detected in the hemolymph and midgut which were the most important immune tissues in silkworm. All the results suggested that BmPGRP-S3 may play an important role in the immune response of silkworm to BmCPV infection and provided helpful information for further studying the function of BmPGRP-S3 in silkworm. PMID:25218236

  1. Identification of a Novel Strong and Ubiquitous Promoter/Enhancer in the Silkworm Bombyx mori

    PubMed Central

    Tsubota, Takuya; Uchino, Keiro; Suzuki, Takao K; Tanaka, Hiromitsu; Kayukawa, Takumi; Shinoda, Tetsuro; Sezutsu, Hideki

    2014-01-01

    Transgenic techniques offer a valuable tool for determining gene functions. Although various promoters are available for use in gene overexpression, gene knockdown, and identification of transgenic individuals, there is nevertheless a lack of versatile promoters for such studies, and this dearth acts as a bottleneck, especially with regard to nonmodel organisms. Here, we succeeded in identifying a novel strong and ubiquitous promoter/enhancer in the silkworm. We identified a unique silkworm strain whose reporter gene showed strong and ubiquitous expression during the establishment of enhancer trap strains. In this strain, the transposon was inserted into the 5?UTR of hsp90, a housekeeping gene that is abundantly expressed in a range of tissues. To determine whether the promoter/enhancer of hsp90 could be used to induce strong gene expression, a 2.9-kb upstream genomic fragment of hsp90 was isolated (hsp90P2.9k), and its transcriptional activation activity was examined. Strikingly, hsp90P2.9k induced strong gene expression in silkworm cell cultures and also strongly induced gene expression in various tissues and developmental stages of the silkworm. hsp90P2.9k also exhibited significant promoter/enhancer activity in Sf9, a cell culture from the armyworm, suggesting that this fragment might possibly be used as a gene expression tool in other Lepidoptera. We further found that 2.0 kb of hsp90P2.9k is sufficient for the induction of strong gene expression. We believe that this element will be of value for a range of studies such as targeted gene overexpression, gene knockdown and marker gene expression, not only in the silkworm but also in other insect species. PMID:24875626

  2. Analysis of genetic relationship in mutant silkworm strains of Bombyx mori using inter simple sequence repeat (ISSR) markers.

    PubMed

    Velu, Dhanikachalam; Ponnuvel, Kangayam M; Muthulakshmi, Murugiah; Sinha, Randhir K; Qadri, Syed M H

    2008-05-01

    Amplified inter simple sequence repeats (ISSR) markers were used to determine genetic relationships among mutant silkworm strains of Bombyx mori. Fifteen ISSR primers containing simple sequence repeat (SSR) motifs were used in this study. A total of 113 markers were produced among 20 mutant strains, of which 73.45% were found to be polymorphic. In selected mutant genetic stocks, the average number of observed allele was (1.7080+/-0.4567), effective alleles (1.5194+/-0.3950) and genetic diversity (Ht) (0.2901+/-0.0415). The dendrogram produced using the unweighted pair group method with arithmetic means (UPGMA) and cluster analysis made using Nei's genetic distance resulted in the formation of one major group containing 6 groups separated 20 mutant silkworm strains. Therefore, ISSR amplification is a valuable method for determining the genetic variability among mutant silkworm strains. This efficient molecular marker would be useful for characterizing a considerable number of silkworm strains maintained at the germplasm center. PMID:18499073

  3. A novel third chromosomal locus controls susceptibility to Autographa californica multiple nucleopolyhedrovirus in the silkworm, Bombyx mori.

    PubMed

    Xu, Jian; Kusakabe, Takahiro; Yamamoto, Kimiko; Suetsugu, Yoshitaka; Mon, Hiroaki; Li, Zhiqing; Zhu, Li; Iiyama, Kazuhiro; Banno, Yutaka; Yoshimura, Kaito; Lee, Jae Man

    2014-04-01

    Baculovirus demonstrates specific infection spectrums and thus one certain host exhibits particular response to single baculovirus isolate. Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is considered to be not an innate pathogen to Bombyx mori, but some silkworm strains have been identified to be permissive to AcMNPV, indicating the positive or negative involvement of certain host factors in baculovirus replications in vivo. To provide a fundamental knowledge of this process, we performed large-scale screening to investigate the responses of 448 silkworm strains against recombinant AcMNPV inoculation. By genetic analysis between permissive and resistant strains identified, we further confirmed that a potential corresponding locus on chromosome 3 regulates host responses to AcMNPV in silkworm. Additionally, we found that it is available for AcMNPV-silkworm baculovirus expression vector system to produce proteins of interest. PMID:24337396

  4. Identification and Expression Profiling of the BTB Domain-Containing Protein Gene Family in the Silkworm, Bombyx mori

    PubMed Central

    Cheng, Daojun; Qian, Wenliang; Meng, Meng; Wang, Yonghu; Peng, Jian; Xia, Qingyou

    2014-01-01

    The BTB domain is a conserved protein-protein interaction motif. In this study, we identified 56 BTB domain-containing protein genes in the silkworm, in addition to 46 in the honey bee, 55 in the red flour beetle, and 53 in the monarch butterfly. Silkworm BTB protein genes were classified into nine subfamilies according to their domain architecture, and most of them could be mapped on the different chromosomes. Phylogenetic analysis suggests that silkworm BTB protein genes may have undergone a duplication event in three subfamilies: BTB-BACK-Kelch, BTB-BACK-PHR, and BTB-FLYWCH. Comparative analysis demonstrated that the orthologs of each of 13 BTB protein genes present a rigorous orthologous relationship in the silkworm and other surveyed insects, indicating conserved functions of these genes during insect evolution. Furthermore, several silkworm BTB protein genes exhibited sex-specific expression in larval tissues or at different stages during metamorphosis. These findings not only contribute to a better understanding of the evolution of insect BTB protein gene families but also provide a basis for further investigation of the functions of BTB protein genes in the silkworm. PMID:24895545

  5. Fluorescence imaging for a noninvasive in vivo toxicity-test using a transgenic silkworm expressing green fluorescent protein.

    PubMed

    Inagaki, Yoshinori; Matsumoto, Yasuhiko; Ishii, Masaki; Uchino, Keiro; Sezutsu, Hideki; Sekimizu, Kazuhisa

    2015-01-01

    In drug development, the toxicity of candidate chemicals must be carefully examined in an animal model. Here we developed a live imaging technique using silkworms for a noninvasive toxicity test applicable for drug screening. Injection of carbon tetrachloride, a tissue-injuring chemical, into transgenic silkworms expressing green fluorescent protein (GFP) induced leakage of GFP from the tissues into the hemolymph. The leakage of GFP was suppressed by pre-administration of either cimetidine, a cytochrome P450 inhibitor, or N-acetyl cysteine, a free-radical scavenger. The transgenic silkworm was made transparent by feeding a diet containing chemicals that inhibit uric acid deposition in the epithelial cells. In the transparent silkworms, GFP fluorescence in the fat body could be observed from outside the body. Injection of salicylic acid or iron sulfate, tissue-injuring chemicals, into the transparent silkworms decreased the fluorescence intensity of the GFP in the fat body. These findings suggest that the transparent GFP-expressing silkworm model is useful for evaluating the toxicity of chemicals that induce tissue injury. PMID:26061948

  6. Fluorescence imaging for a noninvasive in vivo toxicity-test using a transgenic silkworm expressing green fluorescent protein

    PubMed Central

    Inagaki, Yoshinori; Matsumoto, Yasuhiko; Ishii, Masaki; Uchino, Keiro; Sezutsu, Hideki; Sekimizu, Kazuhisa

    2015-01-01

    In drug development, the toxicity of candidate chemicals must be carefully examined in an animal model. Here we developed a live imaging technique using silkworms for a noninvasive toxicity test applicable for drug screening. Injection of carbon tetrachloride, a tissue-injuring chemical, into transgenic silkworms expressing green fluorescent protein (GFP) induced leakage of GFP from the tissues into the hemolymph. The leakage of GFP was suppressed by pre-administration of either cimetidine, a cytochrome P450 inhibitor, or N-acetyl cysteine, a free-radical scavenger. The transgenic silkworm was made transparent by feeding a diet containing chemicals that inhibit uric acid deposition in the epithelial cells. In the transparent silkworms, GFP fluorescence in the fat body could be observed from outside the body. Injection of salicylic acid or iron sulfate, tissue-injuring chemicals, into the transparent silkworms decreased the fluorescence intensity of the GFP in the fat body. These findings suggest that the transparent GFP-expressing silkworm model is useful for evaluating the toxicity of chemicals that induce tissue injury. PMID:26061948

  7. A novel pattern recognition protein of the Chinese oak silkmoth, Antheraea pernyi, is involved in the pro-PO activating system

    PubMed Central

    Xialu, Wang; Jinghai, Zhang; Ying, Chen; Youlei, Ma; Wenjun, Zou; Guoyuan, Ding; Wei, Li; Mingyi, Zhao; Chunfu, Wu; Rong, Zhang

    2013-01-01

    In this paper, we firstly reported a C-type lectin cDNA clone of 1029 bps from the larvae of A. Pernyi (Ap-CTL) using PCR and RACE techniques. The full-length cDNA contains an open reading frame encoding 308 amino acid residues which has two different carbohydrate-recognition domains (CRDs) arranged in tandem. To investigate the biological activities in the innate immunity, recombinant Ap-CTL was expressed in E. coli with a 6-histidine at the amino-terminus (Ap-rCTL). Besides acted as a broad-spectrum recognition protein binding to a wide range of PAMPs and microorganisms, Ap-rCTL also had the ability to recognize and trigger the agglutination of bacteria and fungi. In the proPO activation assay, Ap-rCTL specifically restored the PO activity of hemolymph blocked by anti-Ap-rCTL antibody in the presence of different PAMPs or microorganisms. In summary, Ap-rCTL plays an important role in insect innate immunity as an pattern recognition protein. [BMB Reports 2013; 46(7): 358-363] PMID:23884102

  8. Dumbarton Oaks Electronic Texts

    NSDL National Science Digital Library

    Located in the Georgetown neighborhood of Washington, DC, Dumbarton Oaks was donated to Harvard University in 1940 as gift from Mr. and Mrs. Robert Woods Bliss. Currently, the Dumbarton Oaks Center offers a number of fellowships in its three areas of academic inquiry, which include Byzantine studies, pre-Columbian studies, and landscape architecture. The organization is also well known for their numerous edited volumes, which deal with the aforementioned research topics. Some of those volumes can be downloaded from this site, and cover a wide array of subjects, including pieces of Byzantine-period gardens and works on gender in pre-Hispanic America. Divided into the center's three main research areas, the lists of works contain a brief abstract about each volume and links to each chapter in the work.

  9. 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. PMID:24268277

  10. 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. PMID:19008047

  11. Mapping of major quantitative trait loci for economic traits of silkworm cocoon.

    PubMed

    Lie, Z; Cheng, L; Fang-yin, D; Shou-min, F

    2010-01-01

    The quantitative trait loci (QTLs) associated with cocoon traits in silkworms were mapped in 44 individuals of a backcross of Dazao females with hybrid F(1) males; the hybrid males were from females of inbred C(1)00 strain, which have white cocoons and superior cocoon traits, crossed with males of inbred strain Dazao, which have green cocoons and inferior cocoon traits. Nineteen putative major QTLs of silkworm cocoon traits, five QTLs of whole cocoon weight, four QTLs of cocoon shell weight, six QTLs of pupa weight, and four QTLs of cocoon shell rate were scattered across nine linkage groups. The variances explained by QTLs for whole cocoon weight, cocoon shell weight, pupa weight, and cocoon shell rate were 51.0, 73.69, 51.80, and 59.52%, respectively. The numbers of major QTLs with contributions above 10% for these traits were two, three, two, and four, respectively. PMID:20092037

  12. Identification of a new protein from silkworm pupas by biological mass spectrometry.

    PubMed

    Jiang, Liang; Liu, Qiong; Hu, Decong; Jiang, Liang; Xu, Huibi; Wang, Hongxia

    2005-01-01

    To find out the efficient peptide of selenium-rich silkworm pupas in inducing the apoptosis of hepatoma cells, selenium-containing proteins were isolated and characterized. One of the two major proteins in selenium-rich silkworm pupas was identified to be a new protein by peptide mass fingerprinting on matrix assisted laser desorption ionization - time of flight - mass spectrometry. Amino acid sequences of peptides digested by trypsin from the new protein were determined by capillary liquid chromatography - electrospray ionization - quadrupole/time of flight - mass spectrometry and searched with Mascot in NCBI database. Results showed that the six major peptides from the protein were also new peptides that could not be found in the database up to date. PMID:17281553

  13. Revisiting the specificity of Mamestra brassicae and Antheraea polyphemus pheromone-binding proteins with a fluorescence binding assay.

    PubMed

    Campanacci, V; Krieger, J; Bette, S; Sturgis, J N; Lartigue, A; Cambillau, C; Breer, H; Tegoni, M

    2001-06-01

    Pheromone-binding proteins (PBPs), located in the sensillum lymph of pheromone-responsive antennal hairs, are thought to transport the hydrophobic pheromones to the chemosensory membranes of olfactory neurons. It is currently unclear what role PBPs may play in the recognition and discrimination of species-specific pheromones. We have investigated the binding properties and specificity of PBPs from Mamestra brassicae (MbraPBP1), Antheraea polyphemus (ApolPBP1), Bombyx mori (BmorPBP), and a hexa-mutant of MbraPBP1 (Mbra1-M6), mutated at residues of the internal cavity to mimic that of BmorPBP, using the fluorescence probe 1-aminoanthracene (AMA). AMA binds to MbraPBP1 and ApolPBP1, however, no binding was observed with either BmorPBP or Mbra1-M6. The latter result indicates that relatively limited modifications to the PBP cavity actually interfere with AMA binding, suggesting that AMA binds in the internal cavity. Several pheromones are able to displace AMA from the MbraPBP1- and ApolPBP1-binding sites, without, however, any evidence of specificity for their physiologically relevant pheromones. Moreover, some fatty acids are also able to compete with AMA binding. These findings bring into doubt the currently held belief that all PBPs are specifically tuned to distinct pheromonal compounds. PMID:11274212

  14. Cloning of putative odorant-degrading enzyme and integumental esterase cDNAs from the wild silkmoth, Antheraea polyphemus.

    PubMed

    Ishida, Yuko; Leal, Walter S

    2002-12-01

    Odorant-degrading enzymes have been postulated to participate in the fast deactivation of insect pheromones. These proteins are expressed specifically in the sensillar lymph of insect antennae in such low amounts that, hitherto, isolation and protein-based cDNA cloning has not been possible. Using degenerate primers based on conserved amino acid sequences of insect carboxylesterases and juvenile hormone esterases, we were able to amplify partial cDNA fragments, which were then used for the design of gene-specific primers for RACE. This bioinformatics approach led us to the cloning of cDNAs, encoding a putative odorant-degrading enzyme (Apol-ODE) and a putative integumental esterase (Apol-IE) from the wild silkmoth, Antheraea polyphemus. Apol-ODE had a predicted molecular mass of 59,994 Da, pI of 6.63, three potential N-glycosylation sites, and a putative catalytic site Ser characterized by the sequence Gly(195)-Glu-Ser-Ala-Gly-Ala. Apol-IE gave calculated molecular mass of 61,694 Da, pI of 7.49, two potential N-glycosylation sites, and a putative active site with the sequence Gly(214)-Tyr-Ser-Ala-Gly. The transcript of Apol-ODE was detected by RT-PCR in male antennae and branches (sensillar tissues), but not in female antennae and other control tissues. Apol-IE was detected in male and female antennae as well as legs. PMID:12429129

  15. Pheromone-binding proteins contribute to the activation of olfactory receptor neurons in the silkmoths antheraea polyphemus and Bombyx mori.

    PubMed

    Pophof, Blanka

    2004-02-01

    The sensilla trichodea of the silkmoth Antheraea polyphemus are innervated by three types of receptor neurons each responding specifically to one of three pheromone components. The sensillum lymph of these sensilla surrounding the sensory dendrites contains three different types of pheromone-binding proteins (PBPs) in high concentrations. The sensilla trichodea of the silkmoth Bombyx mori are supplied by two receptor neurons each tuned specifically to one of the two pheromone components bombykol and bombykal, but only one type of PBP has been found so far in these sensilla. Recombinant PBPs of both silkmoth species in various combinations with pheromone components were applied to the receptor neurons via tip-opened sensilla during electrophysiological recordings. Over a fairly broad range of pheromone concentrations the responses of the receptor neurons depended on both, the pheromone component and the type of the PBP. Therefore, the PBPs appear to contribute to the excitation of the receptor neurons. Furthermore, bombykal in combination with the expressed PBP of B. mori failed to activate the corresponding receptor neuron of B. mori, but did so if combined with one of the PBPs of A. polyphemus. Therefore, a still unknown binding protein involved in bombykal transport might be present in B. mori. PMID:14977808

  16. Morphogenesis of the antenna of the male silkmoth, Antheraea polyphemus. VI. Experimental disturbance of antennal branch formation.

    PubMed

    Steiner, C; Keil, T A

    1995-06-01

    In the male silkmoth Antheraea polyphemus, the formation of the side branches of the quadripectinate antennal flagellum was disturbed by an experimental manipulation. Normally the side branches develop in the pupa via deep incisions which proceed from the periphery towards the centerline of the leaf-shaped antennal anlage. Local removal of the uppermost, pigmented cuticular layers of the pupal antennal pocket ('cuticular window') led to a local standstill of branch formation in the manipulated region of the pocket, most probably caused by increased evaporation of water through the remaining layers of meso- and endocuticle. These parts of the antenna retained an unbranched, plate-like shape. This early morphogenetic stage was conserved by the secretion of antennal cuticle. Besides cuticle formation, development of sensilla is not impeded by the manipulation. In the plate-shaped regions, the initial pattern formed by the sensilla in the antennal epidermis is preserved, because they maturate at their birth places. In the individual segments, the pattern of sensilla shows a mirror-like symmetry with respect to the segmental midline. From the edge to the midline, we found large s. trichodea, followed by small s. trichodea, s. basiconica, and s. coeloconica on the dorsal side whereas on the ventral side, there are only large s. trichodea and s. campaniformia. We conclude that the development of the featherlike antennal shape on the one hand and the development of sensilla and cuticle on the other hand are independent processes. PMID:18621301

  17. 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 [Department of Biotechnology, Indian Institute of Technology, Kharagpur, Kharagpur 721302, West Bengal (India); Ghosh, Ananta Kumar, E-mail: aghosh@hijli.iitkgp.ernet.i [Department of Biotechnology, Indian Institute of Technology, Kharagpur, Kharagpur 721302, West Bengal (India)

    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.

  18. Comparative proteomic analysis between the domesticated silkworm (Bombyx mori) reared on fresh mulberry leaves and on artificial diet.

    PubMed

    Zhou, Zhong-Hua; Yang, Hui-Juan; Chen, Ming; Lou, Cheng-Fu; Zhang, Yao-Zhou; Chen, Ke-Ping; Wang, Yong; Yu, Mei-Lan; Yu, Fang; Li, Jian-Ying; Zhong, Bo-Xiong

    2008-12-01

    To gain an insight into the effects of different diets on growth and development of the domesticated silkworm at protein level, we employed comparative proteomic approach to investigate the proteomic differences of midgut, hemolymph, fat body and posterior silk gland of the silkworms reared on fresh mulberry leaves and on artificial diet. Seventy-six differentially expressed proteins were identified by MALDI TOF/TOF MS, and among them, 41 proteins were up-regulated, and 35 proteins were downregulated. Database searches, combined with GO analysis and KEGG pathway analysis revealed that some hemolymph proteins such as Nuecin, Gloverin-like proteins, PGRP, P50 and beta/-N-acetylglucosamidase were related to innate immunity of the silkworm, and some proteins identified in silkworm midgut including Myosin 1 light chain, Tropomyosin 1, Profilin, Serpin-2 and GSH-Px were involved in digestion and nutrition absorption. Moreover, two up-regulated enzymes in fat body of larvae reared on artificial diet were identified as V-ATPase subunit B and Arginine kinase which participate in energy metabolism. Furthermore, 6 down-regulated proteins identified in posterior silk gland of silkworm larvae reared on artificial diet including Ribosomal protein SA, EF-2, EF-1gamma, AspAT, ERp57 and PHB were related to silk synthesis. Our results suggested that the different diets could alter the expression of proteins related to immune system, digestion and absorption of nutrient, energy metabolism and silk synthesis poor nutrition and absorption of nutrition in silkworm. The results also confirmed that the poor nutrient absorption, weakened innate immunity, decreased energy metabolism and reduced silk synthesis are the main reasons for low cocoons yield, inferior filament quality, low survival rate of young larvae and insufficient resistance against specific pathogens in the silkworms fed on artificial diet. PMID:18998723

  19. Mechanical Properties of Regenerated Bombyx mori Silk Fibers and Recombinant Silk Fibers Produced by Transgenic Silkworms

    Microsoft Academic Search

    Zhenghua Zhu; Yuka Kikuchi; Katsura Kojima; Toshiki Tamura; Nobuo Kuwabara; Takashi Nakamura; Tetsuo Asakura

    2010-01-01

    Regenerated silk fibroin fibers from the cocoons of silkworm, Bombyx mori, were prepared with hexafluoro solvents, 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) or hexafluoroacetone-trihydrate (HFA), as dope solvents and methanol as coagulation solvent. The regenerated fiber prepared from the HFIP solution showed slightly larger tensile strength when the draw ratio is 1:3 than that of native silk fiber, but the strength of the regenerated

  20. Genetic variants of the Bombyx mori silkworm encoding sericin proteins of different lengths

    Microsoft Academic Search

    Takuma Gamo

    1982-01-01

    A variant sericin polypeptide originally found by acid gel electrophoresis in the Nd-s mutant strain of the silkworm, Bombyx mori, has been analyzed genetically. The vriant polypeptide (called S-2v) is encoded by a gene which behaves as a codominant allele of the gene encoding the standard S-2 sericin polypeptide. Linkage analysis locates these alleles at 0.0 map unit on chromosome

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

    Microsoft Academic Search

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

    To understand mechanisms for the difference of uptaking and transporting the pigments between the male and female in the silkworm,\\u000a Bombyx mori strain of sex-related fluorescent cocoon, the fluorescent pigments in the midgut lumen, midgut, blood, silk glands and cocoon\\u000a were analyzed with thin-layer chromatography, and showed that fluorescent colors of cocoons consisted with that of blood and\\u000a silk glands.

  2. Tensile behavior and morphology of differently degummed silkworm ( Bombyx mori) cocoon silk fibres

    Microsoft Academic Search

    Ping Jiang; Huifen Liu; Changhe Wang; Lingzhi Wu; Jianguo Huang; Cong Guo

    2006-01-01

    Tensile tests have been performed on silkworm cocoon silk fibres degummed using five different methods: distilled water, boracic acid–sodium borate buffer, sodium carbonate, urea and succinic acid. Using an electronic single-fibre tensile instron, the force–displacement curves were obtained for each condition. Effects of degumming on silk include a decrease in the initial elastic modulus and a decrease in the proportional

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

    Microsoft Academic Search

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

    2008-01-01

    Most animals have the ability to adapt, to some extends and in different ways, the variation or disturbance of environment.\\u000a In our experiments, we forced a silkworm caterpillar to spin two, three or four thin cocoons by taking it out from the cocoon\\u000a being constructed. The mechanical properties of these cocoons were studied by static tensile tests and dynamic mechanical

  4. Transgenic silkworms ( Bombyx mori ) produce recombinant spider dragline silk in cocoons

    Microsoft Academic Search

    Hongxiu Wen; Xiqian Lan; Yuansong Zhang; Tianfu Zhao; Yujun Wang; Zenta Kajiura; Masao Nakagaki

    2010-01-01

    Spider dragline silk is a unique fibrous protein with a combination of tensile strength and elasticity, but the isolation\\u000a of large amounts of silk from spiders is not feasible. In this study, we generated germline-transgenic silkworms (Bombyx mori) that spun cocoons containing recombinant spider silk. A piggyBac-based transformation vector was constructed that carried spider dragline silk (MaSp1) cDNA driven by

  5. A new continuous cell line from larval ovaries of silkworm, Bombyx mori

    Microsoft Academic Search

    Arun M. Khurad; Min-Juan Zhang; Chanchal G. Deshmukh; Ravindra S. Bahekar; Ashish D. Tiple; Chuan-Xi Zhang

    2009-01-01

    A new continuous cell line from ovarian tissue of commercial variety “Kolar Gold” of silkworm, Bombyx mori, was established and designated as DZNU-Bm-12. The tissue was grown in MGM-448 insect cell culture medium supplemented with\\u000a 10% fetal bovine serum (FBS) and 3% heat-inactivated B. mori hemolymph at 25?±?1°C. The migration of partially attached small round refractive cells from the fragments

  6. Phylogenetic relationships of three new microsporidian isolates from the silkworm, Bombyx mori

    Microsoft Academic Search

    S. Nageswara Rao; M. Muthulakshmi; S. Kanginakudru; J. Nagarajua

    2004-01-01

    The pathogenicity, mode of transmission, tissue specificity of infection and the small subunit rRNA (SSU-rRNA) gene sequences of the three new microsporidian isolates from the silkworm Bombyx mori were studied. Out of the three, NIK-2r revealed life cycle features and SSU-rRNA gene sequence similar to Nosema bombycis, suggesting that it is N. bombycis. The other two, NIK-4m and NIK-3h, differed

  7. Chitin in the Silk Gland Ducts of the Spider Nephila edulis and the Silkworm Bombyx mori

    PubMed Central

    Davies, Gwilym J. G.; Knight, David P.; Vollrath, Fritz

    2013-01-01

    Here we report the detection and localisation of chitin in the cuticle of the spinning ducts of both the spider Nephila edulis and the silkworm Bombyx mori. Our observations demonstrate that the duct walls of both animals contain chitin notwithstanding totally independent evolutionary pathways of the systems. We conclude that chitin may well be an essential component for the construction of spinning ducts; we further conclude that in both species chitin may indicate the evolutionary origin of the spinning ducts. PMID:24015298

  8. Expression pattern of immunoglobulin superfamily members in the silkworm, Bombyx mori.

    PubMed

    He, Lei; Cao, Guangli; Huang, Moli; Xue, Renyu; Hu, Xiaolong; Gong, Chengliang

    2014-09-15

    Immunoglobulin superfamily (IgSF) proteins are involved in cell adhesion, cell communication and immune functions. In this study, 152 IgSF genes containing at least one immunoglobulin (Ig) domain were predicted in the Bombyx mori silkworm genome. Of these, 145 were distributed on 25 chromosomes with no genes on chromosomes 16, 18 and 26. Multiple sequence alignments and phylogenetic evolution analysis indicated that IgSFs evolved rapidly. Gene ontology (GO) annotation indicated that IgSF members functioned as cellular components and in molecular functions and biological processes. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis suggested that IgSF proteins were involved in signal transduction, signaling molecules and interaction, and cell communication. Microarray-based expression data showed tissue expression for 136 genes in anterior silkgland, middle silkgland, posterior silkgland, testis, ovary, fat body, midgut, integument, hemocyte, malpighian tubule and head. Expression pattern of IgSF genes in the silkworm ovary and midgut was analyzed by RNA-Seq. Expression of 105 genes was detected in the ovary in strain Dazao. Expression in the midgut was detected for 74 genes in strain Lan5 and 75 genes in strain Ou17. Expression of 34 IgSF genes in the midgut relative to the actin A3 gene was significantly different between strains Lan5 and Ou17. Furthermore, 1 IgSF gene was upregulated and 1 IgSF gene was downregulated in strain Lan5, and 4 IgSF genes were upregulated and 2 IgSF genes were downregulated in strain Ou17 after silkworms were challenged with B. mori cypovirus (BmCPV), indicating potential involvement in the response to BmCPV-infection. These results provide an overview of IgSF family members in silkworms, and lay the foundation for further functional studies. PMID:25020261

  9. Cloning and purification of recombinant silkworm dihydrolipoamide dehydrogenase expressed in Escherichia coli

    Microsoft Academic Search

    Juan Huo; Haifeng Shi; Qin Yao; Huiqin Chen; Lin Wang; Keping Chen

    2010-01-01

    Dihydrolipoamide dehydrogenase (DLDH), a flavin-dependent oxidoreductase is essential for energy metabolism. As an oxidoreductase it catalyzes the NAD+-dependent oxidation of dihydrolipoamide. In this study, a putative Bombyx mori dihydrolipoamide dehydrogenase (BmDLDH) gene was cloned, expressed, purified and characterized for the first time. The BmDLDH gene was amplified from a pool of silkworm cDNAs by PCR and cloned into Escherichia coli

  10. 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 further work on host-parasite interaction between insects and microsporidia. PMID:24386341

  11. MicroRNA expression profiling during the life cycle of the silkworm (Bombyx mori)

    Microsoft Academic Search

    Shiping Liu; Liang Zhang; Qibin Li; Ping Zhao; Jun Duan; Daojun Cheng; Zhonghuai Xiang; Qingyou Xia

    2009-01-01

    BACKGROUND: MicroRNAs (miRNAs) are expressed by a wide range of eukaryotic organisms, and function in diverse biological processes. Numerous miRNAs have been identified in Bombyx mori, but the temporal expression profiles of miRNAs corresponding to each stage transition over the entire life cycle of the silkworm remain to be established. To obtain a comprehensive overview of the correlation between miRNA

  12. 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. PMID:21604173

  13. 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.

  14. 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.

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

    SciTech Connect

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

    2009-07-31

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

  16. Roles of Piwi Proteins in Transcriptional Regulation Mediated by HP1s in Cultured Silkworm Cells

    PubMed Central

    Tatsuke, Tsuneyuki; Zhu, Li; Li, Zhiqing; Mitsunobu, Hitoshi; Yoshimura, Kaito; Mon, Hiroaki; Lee, Jae Man; Kusakabe, Takahiro

    2014-01-01

    Piwi proteins are part of a superfamily of Argonaute proteins, which are one of the core components of the RNA silencing pathway in many eukaryotes. Piwi proteins are thought to repress the transposon expression both transcriptionally and post-transcriptionally. Recently, Drosophila melanogaster Piwi was recently reported to associate with chromatin and to interact directly with the Heterochromatin Protein 1 (HP1a). However, similar interactions have not been reported in other higher eukaryotes. Here we show that silkworm Piwi proteins interact with HP1s in the nucleus. The silkworm, Bombyx mori, has two Piwi proteins, Ago3 and Siwi, and two typical HP1 proteins, HP1a and HP1b. We found that HP1a plays an important role in the interaction between Ago3/Siwi and HP1b in the ovary-derived BmN4 cell line. We also found that Ago3/Siwi regulates the transcription in an HP1-dependent manner. These results suggest that silkworm Piwi proteins function as a chromatin regulator in collaboration with HP1a and HP1b. PMID:24637637

  17. Recombinant canine parvovirus-like particles express foreign epitopes in silkworm pupae.

    PubMed

    Feng, Hao; Liang, Meng; Wang, Hua-lei; Zhang, Tao; Zhao, Ping-sen; Shen, Xing-jia; Zhang, Ren-zhou; Hu, Gui-qiu; Gao, Yu-qei; Wang, Cheng-yu; Wang, Tie-cheng; Zhang, Wei; Yang, Song-tao; Xia, Xian-zhu

    2011-12-29

    The capsid structural protein VP2 of canine parvovirus (CPV) can self-assemble into highly organized virus-like particles (VLPs) and retain major immunoreactivity. In this study, different recombinant baculoviruses that expressed varying fusion proteins of the CPV VP2 protein with the T cell determinant and/or the linear virus-neutralizing epitope of rabies virus (RV) were generated. Infection with these baculoviruses changed BmN cell morphology and inhibited their proliferation as well as damaged silkworms and pupae. However, infection with these baculoviruses induced high levels of recombinant protein expression in silkworms and pupae. More importantly, these fusion proteins self-assembled VLPs with properties similar to CPV virions and retained their VP2-specific immunoreactivity, but some retained their RV-specific immunoreactivity. Interestingly, only one fusion protein, T-VP2, maintained its haemagglutination activity. These data indicated that these insertions and replacements in the loop 2 of VP2 did not interfere with the formation of VLP, and silkworms and pupae could act as a low-costing bioreactor for the production of heterologous proteins. Therefore, our findings may provide a new framework for the development of subunit vaccines against RV and CPV. PMID:21782359

  18. 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

  19. Stable isotope labeling of glycoprotein expressed in silkworms using immunoglobulin G as a test molecule.

    PubMed

    Yagi, Hirokazu; Nakamura, Masatoshi; Yokoyama, Jun; Zhang, Ying; Yamaguchi, Takumi; Kondo, Sachiko; Kobayashi, Jun; Kato, Tatsuya; Park, Enoch Y; Nakazawa, Shiori; Hashii, Noritaka; Kawasaki, Nana; Kato, Koichi

    2015-06-01

    Silkworms serve as promising bioreactors for the production of recombinant proteins, including glycoproteins and membrane proteins, for structural and functional protein analyses. However, lack of methodology for stable isotope labeling has been a major deterrent to using this expression system for nuclear magnetic resonance (NMR) structural biology. Here we developed a metabolic isotope labeling technique using commercially available silkworm larvae. The fifth instar larvae were infected with baculoviruses for co-expression of recombinant human immunoglobulin G (IgG) as a test molecule, with calnexin as a chaperone. They were subsequently reared on an artificial diet containing (15)N-labeled yeast crude protein extract. We harvested 0.1 mg of IgG from larva with a (15)N-enrichment ratio of approximately 80 %. This allowed us to compare NMR spectral data of the Fc fragment cleaved from the silkworm-produced IgG with those of an authentic Fc glycoprotein derived from mammalian cells. Therefore, we successfully demonstrated that our method enables production of isotopically labeled glycoproteins for NMR studies. PMID:25902760

  20. 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 ?-alanine and N-?-alanyl dopamine (NBAD), but accumulation of dopamine. Following injection of ?-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

  1. 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

    2015-01-01

    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 ?-alanine and N-?-alanyl dopamine (NBAD), but accumulation of dopamine. Following injection of ?-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

  2. Oaks Through Time: Reconstructing Historical Change in Oak Landscapes 1

    Microsoft Academic Search

    Arthur Dawson

    2006-01-01

    The San Francisco Bay Area was home to some of the most celebrated valley oak lands in California, their grandeur much chronicled by early visitors and residents. Despite extensive loss, there remains substantial local interest in their preservation and restoration, necessitating an understanding of historical distribution and subsequent change. Assembling and interpreting historical and current oak data for highly modified

  3. 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.

  4. A Review of the Implications of Heterozygosity and Inbreeding on Germplasm Biodiversity and Its Conservation in the Silkworm, Bombyx mori

    PubMed Central

    Jingade, A.H.; Vijayan, K.; Somasundaram, P.; Srivasababu, G.K.; Kamble, C.K.

    2011-01-01

    Silkworm genebanks assume paramount importance as the reservoirs of biodiversity and source of alleles that can be easily retrieved for genetic enhancement of popular breeds. More than 4000 Bombyx mori L (Lepidoptera: Bombycidae) strains are currently available and these strains are maintained through continuous sibling mating. This repeated sibling mating makes the populations of each strain more homozygous, but leads to loss of unique and valuable genes through the process of inbreeding depression. Hence, it is essential to maintain a minimal degree of heterozygosity within the population of each silkworm strain, especially in the traditional geographic strains, to avoid such loss. As a result, accurate estimation of genetic diversity is becoming more important in silkworm genetic resources conservation. Application of molecular markers help estimate genetic diversity much more accurately than that of morphological traits. Since a minimal amount of heterozygosity in each silkworm strain is essential for better conservation by avoiding inbreeding depression, this article overviews both theoretical and practical importance of heterozygosity together with impacts of inbreeding depression and the merits and demerits of neutral molecular markers for measurements of both heterozygosity and inbreeding depression in the silkworm Bombyx mori. PMID:21521139

  5. A review of the implications of heterozygosity and inbreeding on germplasm biodiversity and its conservation in the silkworm, Bombyx mori.

    PubMed

    Jingade, A H; Vijayan, K; Somasundaram, P; Srivasababu, G K; Kamble, C K

    2011-01-01

    Abstract Silkworm genebanks assume paramount importance as the reservoirs of biodiversity and source of alleles that can be easily retrieved for genetic enhancement of popular breeds. More than 4000 Bombyx mori L (Lepidoptera: Bombycidae) strains are currently available and these strains are maintained through continuous sibling mating. This repeated sibling mating makes the populations of each strain more homozygous, but leads to loss of unique and valuable genes through the process of inbreeding depression. Hence, it is essential to maintain a minimal degree of heterozygosity within the population of each silkworm strain, especially in the traditional geographic strains, to avoid such loss. As a result, accurate estimation of genetic diversity is becoming more important in silkworm genetic resources conservation. Application of molecular markers help estimate genetic diversity much more accurately than that of morphological traits. Since a minimal amount of heterozygosity in each silkworm strain is essential for better conservation by avoiding inbreeding depression, this article overviews both theoretical and practical importance of heterozygosity together with impacts of inbreeding depression and the merits and demerits of neutral molecular markers for measurements of both heterozygosity and inbreeding depression in the silkworm Bombyx mori. PMID:21521139

  6. VARIATION IN FLOOD TOLERANCE OF CONTAINER-GROWN SEEDLINGS OF SWAMP WHITE OAK, BUR OAK, AND WHITE OAK

    Microsoft Academic Search

    Michael P. Walsh; J. W. Van Sambeek; Mark V. Coggeshall

    How much variation in flood tolerance exists among seedlings within oak species, given the flood frequency of sites from which acorns are collected, has been largely unexplored. Our studies examined initial growth and flood tolerance for seedlings of swamp white oak (Quercus bicolor Willd.), bur oak (Q. macrocarpa L.), and white oak (Q. alba L.) grown from acorns collected from

  7. Regeneration of the femoral epicondyle on calcium-binding silk scaffolds developed using transgenic silk fibroin produced by transgenic silkworm.

    PubMed

    Nagano, Aya; Tanioka, Yumi; Sakurai, Nobuko; Sezutsu, Hideki; Kuboyama, Noboru; Kiba, Hideo; Tanimoto, Yasuhiro; Nishiyama, Norihiro; Asakura, Tetsuo

    2011-03-01

    Genetically modified silk fibroin containing a poly-glutamic acid site, [(AGSGAG)4E8AS]4, for mineralization was produced as fibers by transgenic silkworms through systematic transformation of the silkworms. The Ca binding activity and mineralization of the transgenic silk fibroin were examined in vitro, showing that this transgenic silk fibroin had relatively high Ca binding activity compared with native silk fibroin. Porous silk scaffolds were prepared with the transgenic and native silk fibroins. Healing of femoral epicondyle defects in rabbit femurs treated with the scaffolds was examined by observing changes in images of the defects using micro-computed tomography. Earlier mineralization and bone formation were observed with scaffolds of transgenic silk fibroin compared with those of native silk fibroin. Thus, this study shows the feasibility of using genetically modified silk fibroin from transgenic silkworms as a mineralization-accelerating material for bone repair. PMID:21055485

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

    PubMed

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

    2013-01-01

    The most important pathogenic fungus of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), is Beauveria bassiana (Balsamo-Crivelli ) Vuillemin (Hypocreales: Clavicipitaceae), which causes significant damage to sericulture production. Therefore, diagnosing fungal disease and developing new control measures are crucial to silk production. To better understand the responsive and interactive mechanisms between the host silkworm and this fungus, variations in silkworm gene expression were investigated using the suppression subtractive hybridization method following the injection of B. bassiana conidia. Two cDNA libraries were constructed, and 140 cDNA clones were isolated. Of the 50 differentially expressed genes identified, 45 (112 clones) were identified in the forward library, and 5 (28 clones) were identified in the reverse library. Expression profiling of six of these genes by quantitative polymerase chain reaction (qPCR) verified that they were induced by the fungal challenge. The present study provides insight into the interaction between lepidopteran insects and pathogenic fungi. PMID:24794288

  9. 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. PMID:25533535

  10. Expression of SNMP-1 in olfactory neurons and sensilla of male and female antennae of the silkmoth Antheraea polyphemus.

    PubMed

    Rogers, M E; Steinbrecht, R A; Vogt, R G

    2001-03-01

    SNMP-1 (sensory neuron membrane protein 1) is an olfactory-specific membrane-bound protein which is homologous with the CD36 receptor family. Previous light level immunocytochemical studies suggested that SNMP-1 was localized in the dendrites and distal cell body of sex-pheromone-specific olfactory receptor neurons (ORN); these studies further suggested SNMP-1 was expressed in only one of two to three neurons in male-specific pheromone-sensitive trichoid sensilla. To better understand the expression and localization of SNMP-1, an immunocytochemical study was performed using electron microscopy to visualize the distribution of SNMP-1 among the neurons of several classes of olfactory sensilla of both male and female antennae of the silkmoth Antheraea polyphemus. SNMP-1 antigenicity was primarily restricted to the receptive dendritic membranes of ORNs of all sensilla types examined and was observed in cytosolic granules, but not plasma membranes, of the cell soma. Mean labeling densities ranged from 1 to 16 gold particles per micrometer of dendrite circumference; dendrites of trichoid and intermediate sensilla showed significantly higher labeling densities than those of basiconic sensilla. Larger dendrites of trichoid sensilla showed significantly higher mean labeling densities (13-16/micron) than smaller diameter dendrites (3-7/micron). Immunofluorescence studies using baculovirus expressed SNMP-1 and multiphoton photon laser scanning microscopy (MPLSM) indicated that rSNMP-1, which was post-translationally processed to the in vivo molecular weight, was inserted into the plasma membrane in a topography presenting extracellular epitopes. These studies suggest SNMP-1 is a common feature of the ORNs, is asymmetrically expressed among functionally distinct neurons, and possesses a topography which permits interaction with components of the extracellular sensillum lymph. PMID:11320659

  11. Morphogenesis of the antenna of the male silkmoth, Antheraea polyphemus. V. Development of the peripheral nervous system.

    PubMed

    Steiner, C; Keil, T A

    1995-06-01

    The imaginal antenna of the male silkmoth Antheraea polyphemus is a feather-shaped structure consisting of about 30 flagellomeres, each of which gives off two pairs of side branches. During the pupal stage (lasting for 3 weeks), the antenna develops from a leaf-shaped, flattened epidermal sac ('antennal blade') via two series of incisions which proceed from the periphery towards the prospective antennal stem. The development of the peripheral nervous system was studied by staining the neurons with an antibody against horseradish peroxidase as well as by electron microscopy. The epithelium is subdivided in segmentally arranged sensillogenic regions alternating with non-sensillogenic regions. Immediately after apolysis, clusters consisting of 5 sensory neurons each and belonging to the prospective sensilla chaetica can be localized at the periphery of the antennal blade in the sensillogenic regions. During the first day following apolysis, the primordia of ca. 70,000 olfactory sensilla arise in the sensillogenic regions. Axons from their neurons are collected in segmentally arranged nerves which run towards the CNS along the dorsal as well as the ventral epidermis and are enveloped by a glial sheath. This 'primary innervation pattern' is completed within the second day after apolysis. A first wave of incisions ('primary incisions') subdivide the antennal blade into segmental 'double branches' without disturbing the innervation pattern. Then a second wave of incisions ('secondary incisions') splits the double branches into single antennal branches. During this process, the segmental nerves and their glial sheaths are disintegrated. The axons are then redistributed into single branch nerves while their glial sheath is reconstituted, forming the 'secondary', or adult, innervation pattern. The epidermis is backed by a basal lamina which is degraded after outgrowth of the axons, but is reconstituted after formation of the single antennal branches. PMID:18621300

  12. The solution NMR structure of Antheraea polyphemus PBP provides new insight into pheromone recognition by pheromone-binding proteins.

    PubMed

    Mohanty, Smita; Zubkov, Sergey; Gronenborn, Angela M

    2004-03-19

    Pheromone-binding proteins (PBPs) located in the antennae of male moth species play an important role in olfaction. They are carrier proteins, believed to transport volatile hydrophobic pheromone molecules across the aqueous sensillar lymph to the membrane-bound G protein-coupled olfactory receptor proteins. The roles of PBPs in molecular recognition and the mechanisms of pheromone binding and release are poorly understood. Here, we report the NMR structure of a PBP from the giant silk moth Antheraea polyphemus. This is the first structure of a PBP with specific acetate-binding function in vivo. The protein consists of nine alpha-helices: alpha1a (residues 2-5), alpha1b (8-12), alpha1c (16-23), alpha2 (27-34), alpha3a (46-52), alpha3b (54-59), alpha4 (70-79), alpha5 (84-100) and alpha6 (107-125), held together by three disulfide bridges: 19-54, 50-108 and 97-117. A large hydrophobic cavity is located inside the protein, lined with side-chains from all nine helices. The acetate-binding site is located at the narrow end of the cavity formed by the helices alpha3b and alpha4. The pheromone can enter this cavity through an opening between the helix alpha1a, the C-terminal end of the helix alpha6, and the loop between alpha2 and alpha3a. We suggest that Trp37 may play an important role in the initial interaction with the ligand. Our analysis also shows that Asn53 plays the key role in recognition of acetate pheromones specifically, while Phe12, Phe36, Trp37, Phe76, and Phe118 are responsible for non-specific binding, and Leu8 and Ser9 may play a role in ligand chain length recognition. PMID:15003458

  13. 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

  14. Preparation and characterization of Antheraea assama silk fibroin based novel non-woven scaffold for tissue engineering applications.

    PubMed

    Kasoju, Naresh; Bhonde, Ramesh R; Bora, Utpal

    2009-10-01

    The quest for novel materials as scaffolds with suitable micro-architecture for supporting tissue neogenesis in tissue engineering and regenerative medicine (TERM) is continuing. In this paper we report an Antheraea assama silk-based non-woven fibroin scaffold for applications in TERM. The novel three-dimensional scaffold is highly interconnected and porous, with a pore size of 150 microm, porosity of 90% and water uptake capacity of 85%. FTIR revealed a typical beta-sheet structure of fibroin. The scaffold has thermal and mechanical properties superior to those of Bombyx mori, as revealed by DSC, TGA and tensile tests. The scaffold exhibited satisfactory blood compatibility, as determined by thrombogenicity, haemolysis, platelet/leukocyte count, platelet adhesion and protein adsorption studies. The scaffold was found to be cytocompatible with human cell lines A549, KB, HepG2 and HeLa for a period of up to 4 weeks. SEM analysis revealed excellent attachment, spreading and migration of cells in the scaffold. MTT assay was performed to estimate the viability and growth of cells in the matrix. Quantification of collagen in cell-scaffold constructs was done by picro-Sirius red assay. Ex ovo chorioallantoic membrane assay and nitric oxide estimations in spent culture medium showed the scaffold's ability to promote angiogenesis. Finally, the biodegradability of the scaffold was determined by the weight loss observed upon treatment with trypsin over a period of 4 weeks. The results reveal that the fibroin from A. assama is a promising candidate as a biocompatible, biomimetic and biodegradable biomaterial of natural origin for applications in TERM. PMID:19670334

  15. Reducing blood glucose level in TIDM mice by orally administering the silk glands of transgenic hIGF-I silkworms.

    PubMed

    Cong, Liang; Cao, Guangli; Renyu, Xue; Zhonghua, Pan; Xiaojian, Zheng; Zhou, Wenlin; Gong, Chengliang

    2011-07-15

    To realize the secretory expression of human insulin-like growth factor-I (hIGF-I) in the posterior silk glands (PSGs) of transgenic silkworms, the piggyBac transposon vector pigA3GFP-fibHS-hIGF-i.e.-neo containing a neomycin-resistance gene (neo), green fluorescent protein gene (gfp) and human insulin-like growth factor I (hIGF-I) gene controlled by the Bombyxmori fibroin heavy chain gene (fib-H) promoter with its downstream signal peptide sequence, and a helper plasmid containing the piggyBac transposase sequence under the control of the B. mori actin 3 gene (A3) promoter were transferred into silkworm eggs by sperm-mediated gene transfer. Transformed silkworms were obtained after being screened for green fluorescence and by the antibiotic G418. In the PSGs of the transformed silkworms, a specific band representing hIGF-I could be detected by Western blotting, and the content of the hIGF-I estimated by ELISA was approximately 1.84 ?g/gram of cocoon and 19.18 ?g/gram of freeze-dried PSG powder. To further estimate the biological activity of the expressed hIGF-I, streptozotocin-induced TIDM mice were orally administered with the PSG powder of the transgenic silkworms, the results showed the blood glucose levels of mice were significantly reduced, suggesting that the the PSGs powder of transgenic hIGF-I silkworms could possibly be used as a perorally administered medicine. PMID:21672515

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

    PubMed

    Nie, Hongyi; Liu, Chun; Cheng, Tingcai; Li, Qiongyan; Wu, Yuqian; Zhou, Mengting; Zhang, Yinxia; Xia, Qingyou

    2014-01-01

    In the silkworm Bombyx mori, pigment mutants with diverse body colors have been maintained throughout domestication for about 5000 years. The silkworm larval body color is formed through the mutual interaction of melanin, ommochromes, pteridines and uric acid. These pigments/compounds are synthesized by the cooperative action of various genes and enzymes. Previous reports showed that melanin, ommochrome and pteridine are increased in silkworm quail (q) mutants. To understand the pigment increase and alterations in pigment synthesis in q mutant, transcriptome profiles of the silkworm integument were investigated at 16 h after head capsule slippage in the fourth molt in q mutants and wild-type (Dazao). Compared to the wild-type, 1161 genes were differentially expressed in the q mutant. Of these modulated genes, 62.4% (725 genes) were upregulated and 37.6% (436 genes) were downregulated in the q mutant. The molecular function of differently expressed genes was analyzed by Blast2GO. The results showed that upregulated genes were mainly involved in protein binding, small molecule binding, transferase activity, nucleic acid binding, specific DNA-binding transcription factor activity and chromatin binding, while exclusively down-expressed genes functioned in oxidoreductase activity, cofactor binding, tetrapyrrole binding, peroxidase activity and pigment binding. We focused on genes related to melanin, pteridine and ommochrome biosynthesis; transport of uric acid; and juvenile hormone metabolism because of their importance in integument coloration during molting. This study identified differently expressed genes implicated in silkworm integument formation and pigmentation using silkworm q mutant. The results estimated the number and types of genes that drive new integument formation. PMID:24718369

  17. Proceedings of the Sudden Oak Death Third Science Symposium Welcome to the Sudden Oak Death Third

    E-print Network

    Standiford, Richard B.

    Proceedings of the Sudden Oak Death Third Science Symposium 3 Welcome to the Sudden Oak Death Third to welcome you to the Sudden Oak Death Third Science Symposium. Looking back at the first sudden oak death was first isolated and identified as the causal agent of sudden oak death. It was the summer of 2000

  18. Fine mapping of E(kp)-1, a locus associated with silkworm (Bombyx mori) proleg development.

    PubMed

    Xiang, H; Li, M; Yang, F; Guo, Q; Zhan, S; Lin, H; Miao, X; Huang, Y

    2008-05-01

    The silkworm homeotic mutant E(kp) has a pair of rudimentary abdominal legs, called prolegs, in its A2 segment. This phenotype is caused by a single dominant mutation at the E(kp)-1 locus, which was previously mapped to chromosome 6. To explore the possible association of Hox genes with proleg development in the silkworm, a map-based cloning strategy was used to isolate the E(kp)-1 locus. Five E(kp)-1-linked simple sequence repeat markers on chromosome 6 were used to generate a low-resolution map with a total genetic distance of 39.5 cM. Four additional cleaved amplified polymorphic sequence markers were developed based on the initial map. The closest marker to E(kp)-1 was at a genetic distance of 2.7 cM. A high-resolution genetic map was constructed using nine BC1 segregating populations consisting of 2396 individuals. Recombination suppression was observed in the vicinity of E(kp)-1. Four molecular markers were tightly linked to E(kp)-1, and three were clustered with it. These markers were used to screen a BAC library. A single bacterial artificial chromosome (BAC) clone spanning the E(kp)-1 locus was identified, and E(kp)-1 was delimited to a region less than 220 kb long that included the Hox gene abdominal-A and a non-coding locus, iab-4. These results provide essential information for the isolation of this locus, which may shed light on the mechanism of proleg development in the silkworm and possibly in Lepidoptera. PMID:18364737

  19. Systematic cloning and analysis of autophagy-related genes from the silkworm Bombyx mori

    PubMed Central

    Zhang, Xuan; Hu, Zhan-Ying; Li, Wei-Fang; Li, Qing-Rong; Deng, Xiao-Juan; Yang, Wan-Ying; Cao, Yang; Zhou, Cong-Zhao

    2009-01-01

    Background Through the whole life of eukaryotes, autophagy plays an important role in various biological events including development, differentiation and determination of lifespan. A full set of genes and their encoded proteins of this evolutionarily conserved pathway have been identified in many eukaryotic organisms from yeast to mammals. However, this pathway in the insect model organism, the silkworm Bombyx mori, remains poorly investigated. Results Based on the autophagy pathway in several model organisms and a series of bioinformatic analyses, we have found more than 20 autophagy-related genes from the current database of the silkworm Bombyx mori. These genes could be further classified into the signal transduction pathway and two ubiquitin-like pathways. Using the mRNA extracted from the silkgland, we cloned the full length cDNA fragments of some key genes via reverse transcription PCR and 3' rapid amplification of cDNA ends (RACE). In addition, we found that the transcription levels of two indicator genes BmATG8 and BmATG12 in the silkgland tend to be increased from 1st to 8th day of the fifth instar larvae. Conclusion Bioinformatics in combination with RT-PCR enable us to remodel a preliminary pathway of autophagy in the silkworm. Amplification and cloning of most autophagy-related genes from the silkgland indicated autophagy is indeed an activated process. Furthermore, the time-course transcriptional profiles of BmATG8 and BmATG12 revealed that both genes are up-regulated along the maturation of the silkgland during the fifth instar. These findings suggest that the autophagy should play an important role in Bombyx mori silkgland. PMID:19470186

  20. 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%.

  1. Poison Ivy, Oak, and Sumac

    MedlinePLUS

    ... Z Diseases and treatments M - P Poison ivy Poison ivy, oak, and sumac Rash from poison ivy. ... to an emergency room immediately. Learn more about poison ivy: Poison ivy: Signs and symptoms Poison ivy: ...

  2. Molecular size of the diapause hormone of the silkworm Bombyx mori.

    PubMed

    Kubota, I; Isobe, M; Goto, T; Hasegawa, K

    1976-01-01

    The diapause hormone (DH) responsible for arrested development of the silkworm Bombix mori consists of two active principles of peptide (A and B). Since both hormones form aggregates in aqueous phases, the molecular weights were determined by gel permeation chromatography of Merckogel OR 6000 using methanol-dichloromethane mixture as the developing agent. Gramicidins and modified peptides soluble in the organic solvents were used as the standard markers for the molecular weight measurements of the hormones. The molecular weights of DH-A and -B are estimated to be 3300 +/- 400 and 2000 +/- 200, respectively. PMID:134554

  3. Integration of molecular and classical linkage groups of the silkworm, Bombyx mori (n = 28).

    PubMed

    Yasukochi, Yuji; Banno, Yutaka; Yamamoto, Kohji; Goldsmith, Marian R; Fujii, Hiroshi

    2005-08-01

    Previously published linkage groups (LGs) composed of molecular markers were assigned to classical LGs in the silkworm, Bombyx mori (n = 28). Four markers from the classical linkage map, og, w-1, Lp, and Pfl, were assigned to the molecular linkage maps using sequence tagged sites. In addition, linkage analysis was carried out using BF1 progeny between wild-type and mutant stocks carrying morphological phenotypic markers. As a result, the counterparts for 26 of 28 molecular LGs were identified with their counterparts of the classical LGs. Two visible markers, Sel and Xan, representing different classical LGs, were found to be linked. PMID:16094430

  4. Integrated management of the uzi fly, Exorista bombycis (Louis) (Diptera: Tachinidae), a parasitoid of the silkworm, Bombyx mori L

    Microsoft Academic Search

    Pradip Kumar; D. Manjunath; K. Sathya Prasad; Ram Kishore; Vinod Kumar; R. K. Datta

    1993-01-01

    Exorista bombycis (Louis), known as uzi fly, causes considerable damage to silkworm rearings in several silkproducing countries including India. Various control measures have been adopted by farmers to control uzi fly. Although these measures help to reduce uzi infestation, the economic loss is not completely avoided. We therefore undertook an IPM programme involving spraying of an insecticide (a liquid formulation

  5. 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

  6. Development of a multiplex polymerase chain reaction for the simultaneous detection of microsporidians, nucleopolyhedrovirus, and densovirus infecting silkworms

    Microsoft Academic Search

    G. Ravikumar; S. Raje Urs; N. B. Vijaya Prakash; C. G. P. Rao; K. V. Vardhana

    2011-01-01

    We have developed a novel PCR-based assay for individual and simultaneous detection of three major pathogens (microsporidians, nucleopolyhedrovirus (NPV) and densovirus (DNV)) infecting the silkworm, Bombyx mori. Multiplex PCR, using three primer pairs, two of which were designed from the conserved regions of 16S small subunit ribosomal RNA gene of microsporidians, and polyhedrin gene of NPVs respectively, and a third

  7. Overexpression of recombinant infectious bursal disease virus (IBDV) capsid protein VP2 in the middle silk gland of transgenic silkworm.

    PubMed

    Xu, Hanfu; Yuan, Lin; Wang, Feng; Wang, Yuancheng; Wang, Riyuan; Song, Chunnuan; Xia, Qingyou; Zhao, Ping

    2014-10-01

    Infectious bursal disease virus (IBDV) is the causative agent of a highly contagious disease affecting young chickens and causes serious economic losses to the poultry industry worldwide. Development of subunit vaccine using its major caspid protein, VP2, is one of the promising strategies to protect against IBDV. This study aim to test the feasibility of using silkworm to produce recombinant VP2 protein (rVP2) derived from a very virulent strain of IBDV (vvIBDV). A total of 16 transgenic silkworm lines harboring a codon-optimized VP2 gene driven by the sericin1 promoter were generated and analyzed. The results showed that the rVP2 was synthesized in the middle silk gland of all lines and secreted into their cocoons. The content of rVP2 in the cocoon of each line was ranged from 0.07 to 16.10 % of the total soluble proteins. The rVP2 was purified from 30 g cocoon powders with a yield of 3.33 mg and a purity >90 %. Further analysis indicated that the rVP2 was able to tolerate high temperatures up to 80 °C, and exhibited specific immunogenic activity in mice. To our knowledge, this is the first report of overexpressing rVP2 in the middle silk gland of transgenic silkworm, which demonstrates the capability of silkworm as an efficient tool to produce recombinant immunogens for use in new vaccines against animal diseases. PMID:25106848

  8. Effects of BmKIT3R gene transfer on the development and survival of silkworm Bombyx mori.

    PubMed

    Wang, Xiaojuan; Cao, Guangli; Xue, Renyu; Xie, Min; Zhang, Xing; Yu, Xiaohua; Gong, Chengliang

    2011-12-01

    To verify the effects of gain-of-function mutation of the BmKIT3(R) gene (from the Chinese scorpion Buthus martensii Karsch) on the development and survival rate of insects and to explore a novel strategy for pest control, the effects of BmKIT3(R) gene transfer on the development and survival rate of silkworms were investigated. A novel transgenic vector derived from the piggyBac transposon with the BmKIT3(R) gene controlled by the Bmhsp20.4 promoter was transferred into silkworm eggs. Transgenic silkworms were obtained after screening with GFP and G418 antibiotics and verification by PCR and dot hybridization. The results showed that the oviposition number decreased by 18.9%, and the hatching and final survival rates were approximately 63% and 47.5%, respectively. Some 18.9% of surviving pupae died before developing into moths in the G3 generation. A specific band corresponding to BmKIT3(R) was detected for transgenic silkworms by Western blotting. This indicates that the Bmhsp 20.4 promoter has constitutive expression activity. The significant decrease in the survival rate suggests that pest population numbers could be effectively controlled by using BmKIT3(R) gene transfer. Furthermore, it can be speculated that pupal development to moths could be blocked if BmKIT3(R) were specially expressed in the pupal stage and reeling with fresh cocoons was performed. PMID:21890407

  9. Comparison of multilocus RFLPs and PCR-based marker systems for genetic analysis of the silkworm, Bombyx mori.

    PubMed

    Nagaraju, J; Reddy, K D; Nagaraja, G M; Sethuraman, B N

    2001-05-01

    The utility of multilocus RFLPs and three PCR-based techniques, Random Amplified Polymorphic DNA (RAPD), Inter-Simple Sequence Repeat-PCR (ISSR-PCR) and simple sequence repeats (SSRs) for genetic characterization was examined using 13 diverse silkworm strains. All four approaches successfully discriminated the 13 silkworm varieties but differed in the amount of polymorphism detected. The usefulness of each system was examined in terms of number of loci revealed (effective multiplex ratio, EMR) and the amount of polymorphism detected (diversity index, DI). For example, the six multilocus RFLP probes produced 180 products of which 97% were polymorphic; 15 SSR loci gave rise to an average of 8 alleles each, of which 86% were polymorphic. The ISSR-PCR produced 39 fragments of which 76.98% were polymorphic. The highest diversity index was observed for ISSR-PCR (0.957) and the lowest for RAPDs (0.744). The RAPD, ISSR-PCR and RFLP assays clearly separated the diapausing and non-diapausing silkworm varieties. These results are discussed in terms of choice of appropriate marker technology for different aspects of silkworm genome analysis. PMID:11554975

  10. Genetic mapping of Z chromosome and identification of W chromosome-specific markers in the silkworm, Bombyx mori.

    PubMed

    Nagaraja, G M; Mahesh, G; Satish, V; Madhu, M; Muthulakshmi, M; Nagaraju, J

    2005-08-01

    In the silkworm, Bombyx mori, the female is the heterogametic (ZW) sex and the male is homogametic (ZZ). The female heterogamety is a typical situation in the insect order Lepidoptera. Although the W chromosome in silkworm is strongly female determining, no W-linked gene for a morphological character has been found on it. The Z chromosome carries important traits of economic value as well as genes for various phenotypic traits, but only 2% of molecular information based on its relative size is known. Studies conducted so far indicate that the Z-linked genes are not dosage compensated. In the present study, we constructed a genetic map of randomly amplified polymorphic DNA fragments (RAPD), simple sequence repeats (SSR), and fluorescent intersimple sequence repeat PCR (FISSR) markers for the Z chromosome using a backcross mapping population. A total of 16 Z-linked markers were identified, characterized, and mapped using od, a recessive trait for translucent skin as an anchor marker yielding a total recombination map of 334.5 cM. The linkage distances obtained suggested that the markers were distributed throughout the Z chromosome. Four RAPD and four SSR markers that were linked to W chromosome were also identified. The proposed mapping approach should be useful to identify and map sex-linked traits in the silkworm. The economic and evolutionary significance of Z- and W-linked genes in silkworm, in particular, and lepidopterans, in general, is discussed. PMID:15931240

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

    SciTech Connect

    Kurihara, H. [Toray Industries, Inc., New Frontiers Research Laboratories, 1111 Tebiro, Kamakura, Kanagawa 248-8555 (Japan)]. E-mail: Hiroyuki_Kurihara@nts.toray.co.jp; Sezutsu, H. [Transgenic Silkworm Research Center, National Institute of Agrobiological Sciences, 1-2 Owashi, Tsukuba, Ibaraki 305-8634 (Japan); Tamura, T. [Transgenic Silkworm Research Center, National Institute of Agrobiological Sciences, 1-2 Owashi, Tsukuba, Ibaraki 305-8634 (Japan); Yamada, K. [Toray Industries, Inc., New Frontiers Research Laboratories, 1111 Tebiro, Kamakura, Kanagawa 248-8555 (Japan)

    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.

  12. 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. PMID:25739894

  13. PHYSIOLOGY OF INSECT DIAPAUSE. IV. THE BRAIN AND PROTHORACIC GLANDS AS AN ENDOCRINE SYSTEM IN THE CECROPIA SILKWORM

    Microsoft Academic Search

    CARROLL M. WILLIAMS

    In the previous papers of this series (Williams, 1946b, 1947, 1948a) an endo crine basis was described for the production and termination of pupal diapause in the Cecropia silkworm. The onset of diapause was correlated with a temporary failure of the brain in secreting a hormone required for the initiation of adult development. The ultimate release of this “¿?brain hormone”

  14. 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 chlorpyrifos. Overall, our results indicate that a complex expression regulation of Prxs that play important roles in maintaining redox equilibrium state of silkworm to reduce oxidative damage caused by pesticide. PMID:25175646

  15. 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. PMID:24980060

  16. Stress induction of Bm1 RNA in silkworm larvae: SINEs, an unusual class of stress genes

    PubMed Central

    Kimura, Richard H.; Choudary, Prabhakara V.; Stone, Koni K.; Schmid, Carl W.

    2001-01-01

    This study surveys the induction of RNA polymerase III (Pol III)–directed expression of short interspersed element (SINE) transcripts by various stresses in an animal model, silkworm larvae. Sublethal heat shock and exposure to several toxic compounds increase the level of Bm1 RNA, the silkworm SINE transcript, while also transiently increasing expression of a well-characterized stress-induced transcript, Hsp70 messenger RNA (mRNA). In certain cases, the Bm1 RNA response coincides with that of Hsp70 mRNA, but more often Bm1 RNA responds later in recovery. Baculovirus infection and exposure to certain toxic compounds increase Bm1 RNA but not Hsp70 mRNA, showing that SINE induction is not necessarily coupled to transcription of this particular heat shock gene. SINEs behave as an additional class of stress-inducible genes in living animals but are unusual as stress genes because of their high copy number, genomic dispersion, and Pol III–directed transcription. PMID:11599568

  17. Identification of ISSR markers associated with productivity traits in silkworm, Bombyx moni L.

    PubMed

    Chatterjee, S N; Mohandas, T P

    2003-06-01

    Bombyx mori L., commonly recognised around the world as the mulberry silkworm, is characterized by a wide variability in yield and developmental traits, which have been proven through conventional genetic analysis to be of polygenic nature. A large number of morpho-biochemical traits and RFLP and RAPD markers are mapped on different linkage groups, but to this point very little attention has been given to unravelling the genetics of yield traits. To address this issue, polymorphic profiles of 147 markers generated with 12 ISSR primers on the genomic DNA of 20 silkworm stocks of diverse yield status were subjected to multiple regression and discriminant function analyses (DFA). This led to the identification of eight markers generated by six primers, which demonstrated high beta-coefficient indices of -0.451 to -0.940. Furthermore, a significant difference between the yield traits for stocks with and without the specific marker could also be established. The inheritance pattern of one marker, L13800bp, identified at the first step of selection of markers through stepwise regression analyses for five yield parameters is discussed in the context of applying multiple regression analysis for establishing association, if not linkage, between a group of DNA markers and a particular yield trait of polygenic nature and using such markers in molecular marker-assisted breeding programs. PMID:12834060

  18. Existence of prophenoloxidase in wing discs: a source of plasma prophenoloxidase in the silkworm, Bombyx mori.

    PubMed

    Diao, Yupu; Lu, Anrui; Yang, Bing; Hu, Wenli; Peng, Qing; Ling, Qing-Zhi; Beerntsen, Brenda T; Söderhäll, Kenneth; Ling, Erjun

    2012-01-01

    In insects, hemocytes are considered as the only source of plasma prophenoloxidase (PPO). PPO also exists in the hemocytes of the hematopoietic organ that is connected to the wing disc of Bombyx mori. It is unknown whether there are other cells or tissues that can produce PPO and release it into the hemolymph besides circulating hemocytes. In this study, we use the silkworm as a model to explore this possibility. Through tissue staining and biochemical assays, we found that wing discs contain PPO that can be released into the culture medium in vitro. An in situ assay showed that some cells in the cavity of wing discs have PPO1 and PPO2 mRNA. We conclude that the hematopoietic organ may wrongly release hemocytes into wing discs since they are connected through many tubes as repost in previous paper. In wing discs, the infiltrating hemocytes produce and release PPO probably through cell lysis and the PPO is later transported into hemolymph. Therefore, this might be another source of plasma PPO in the silkworm: some infiltrated hemocytes sourced from the hematopoietic organ release PPO via wing discs. PMID:22848488

  19. 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

  20. Efficient and cost effective production of active-form human PKB using silkworm larvae

    PubMed Central

    Maesaki, Ryoko; Satoh, Ryosuke; Taoka, Masato; Kanaba, Teppei; Asano, Tsunaki; Fujita, Chiharu; Fujiwara, Toshinobu; Ito, Yutaka; Isobe, Toshiaki; Hakoshima, Toshio; Maenaka, Katsumi; Mishima, Masaki

    2014-01-01

    Protein kinase B (PKB) also known as Akt is involved in many signal transduction pathways. As alterations of the PKB pathway are found in a number of human malignancies, PKB is considered an important drug target for cancer therapy. However, production of sufficient amounts of active PKB for biochemical and structural studies is very costly because of the necessity of using a higher organism expression system to obtain phosphorylated PKB. Here, we report efficient production of active PKB? using the BmNPV bacmid expression system with silkworm larvae. Following direct injection of bacmid DNA, recombinant PKB? protein was highly expressed in the fat bodies of larvae, and could be purified using a GST-tag and then cleaved. A final yield of approximately 1?mg PKB?/20 larvae was recorded. Kinase assays showed that the recombinant PKB? possessed high phosphorylation activity. We further confirmed phosphorylation on the activation loop by mass spectrometric analysis. Our results indicate that the silkworm expression system is of value for preparation of active-form PKB? with phosphorylation on the activation loop. This efficient production of the active protein will facilitate further biochemical and structural studies and stimulate subsequent drug development. PMID:25125290

  1. Determination of iminosugars in mulberry leaves and silkworms using hydrophilic interaction chromatography-tandem mass spectrometry.

    PubMed

    Nakagawa, Kiyotaka; Ogawa, Kenta; Higuchi, Ohki; Kimura, Toshiyuki; Miyazawa, Teruo; Hori, Masatoshi

    2010-09-15

    Mulberry 1-deoxynojirimycin (DNJ, a potent alpha-glycosidase inhibitor) has been investigated thoroughly for its analytical methods and therapeutic potential against diabetes, whereas little attention has been given to other iminosugars such as 2-O-alpha-D-galactopyranosyl-DNJ (GAL-DNJ) and fagomine. For instance, concentration and composition of these iminosugars in mulberry leaves as well as sericulture products have not been fully characterized due to lack of suitable analytical methods. Here we developed a simultaneous determination method for DNJ, GAL-DNJ, and fagomine using hydrophilic interaction chromatography (HILIC) with tandem mass spectrometry (MS/MS). When mulberry leaf extracts were subjected to HILIC-MS/MS with multiple reaction monitoring (MRM), individual iminosugars could be separated and detected. The developed method is sufficiently sensitive for determining iminosugars in mulberry leaves as well as silkworms, providing new information (e.g., different amounts of iminosugars in mulberry leaf varieties; high DNJ and low GAL-DNJ in the silkworm body, especially in the blood) that is useful for producing iminosugar-rich products for nutraceutical purposes. PMID:20470744

  2. TIL-type protease inhibitors may be used as targeted resistance factors to enhance silkworm defenses against invasive fungi.

    PubMed

    Li, Youshan; Zhao, Ping; Liu, Huawei; Guo, Xiaomeng; He, Huawei; Zhu, Rui; Xiang, Zhonghuai; Xia, Qingyou

    2015-02-01

    Entomopathogenic fungi penetrate the insect cuticle using their abundant hydrolases. These hydrolases, which include cuticle-degrading proteases and chitinases, are important virulence factors. Our recent findings suggest that many serine protease inhibitors, especially TIL-type protease inhibitors, are involved in insect resistance to pathogenic microorganisms. To clarify the molecular mechanism underlying this resistance to entomopathogenic fungi and identify novel genes to improve the silkworm antifungal capacity, we conducted an in-depth study of serine protease inhibitors. Here, we cloned and expressed a novel silkworm TIL-type protease inhibitor, BmSPI39. In activity assays, BmSPI39 potently inhibited the virulence protease CDEP-1 of Beauveria bassiana, suggesting that it might suppress the fungal penetration of the silkworm integument by inhibiting the cuticle-degrading proteases secreted by the fungus. Phenol oxidase activation studies showed that melanization is involved in the insect immune response to fungal invasion, and that fungus-induced excessive melanization is suppressed by BmSPI39 by inhibiting the fungal cuticle-degrading proteases. To better understand the mechanism involved in the inhibition of fungal virulence by protease inhibitors, their effects on the germination of B. bassiana conidia was examined. BmSPI38 and BmSPI39 significantly inhibited the germination of B. bassiana conidia. Survival assays showed that BmSPI38 and BmSPI39 markedly improved the survival rates of silkworms, and can therefore be used as targeted resistance proteins in the silkworm. These results provided new insight into the molecular mechanisms whereby insect protease inhibitors confer resistance against entomopathogenic fungi, suggesting their potential application in medicinal or agricultural fields. PMID:25453359

  3. Oak Tree Preservation in Thousand Oaks, California1 William F. Elmendorf2

    E-print Network

    Standiford, Richard B.

    Oak Tree Preservation in Thousand Oaks, California1 William F. Elmendorf2 Abstract: The City- sake, the oak tree. First adopted in 1972 as an Emergency City Council Proclamation, the City's Oak Tree Ordinance has been considered by some, to be one of the first and toughest municipal native tree

  4. Proceedings of the Sudden Oak Death Third Science Symposium Quantification of Sudden Oak Death

    E-print Network

    Standiford, Richard B.

    Proceedings of the Sudden Oak Death Third Science Symposium 49 Quantification of Sudden Oak Death ramorum, the plant pathogen causing the forest disease known as sudden oak death. First confirmed in 2000 tree that exhibited spectral characteristics of trees killed by sudden oak death within each host

  5. Where Has All the White Oak Gone?

    NSDL National Science Digital Library

    MARC D. ABRAMS (; )

    2003-10-01

    This peer-reviewed article from BioScience is about the disappearance of white oaks in the US. Before European settlement, vast areas of deciduous forest in what is now the eastern United States were dominated by oak species. Among these species, white oak (Quercus alba) reigned supreme. White oak tended to grow at lower elevations but was distributed across a broad range of sites, from wet mesic to subxeric, and grew on all but the wettest and most xeric, rocky, or nutrient-poor soils. A comparison of witness tree data from early land surveys and data on modern-day forest composition revealed a drastic decline in white oak throughout the eastern forest. By contrast, other dominant oaks, such as red oak (Quercus rubra) and chestnut oak (Quercus prinus), often exhibited higher frequency in recent studies than in surveys of the original forest. The frequency of red oak witness trees before European settlement was quite low, generally under 5% in most forests. Red oak's distribution was apparently limited by a lower tolerance to fire and drought and a greater dependence on catastrophic disturbances than that of white oak. During the late 19th and early 20th centuries, much of the eastern forest was decimated by land clearing, extensive clear-cutting, catastrophic fires, chestnut blight, and then fire suppression and intensive deer browsing. These activities had the greatest negative impact on the highly valued white oak, while promoting the expansion of red oak and chestnut oak. More recently, however, recruitment of all the dominant upland oaks has been limited on all but the most xeric sites. Thus, the dynamic equilibrium in the ecology of upland oaks that existed for thousands of years has been destroyed in the few centuries following European settlement.

  6. Small Business Manager Oak Ridge National Laboratory

    E-print Network

    Keith Joy Small Business Manager Oak Ridge National Laboratory: Past, Present, and Future #12;2 OAK RIDGE NATIONAL LABORATORY U. S. DEPARTMENT OF ENERGY Overview_0604 ORNL in 1943 The Clinton Pile was the world's first continuously operated nuclear reactor Oak Ridge National Laboratory evolved from

  7. Deer Oaks EAP Services Fact Sheet

    E-print Network

    Velev, Orlin D.

    Deer Oaks EAP Services Fact Sheet www.deeroaks.com · 1-866-327-2400 · eap@deeroaks.com Eligibility. The Deer Oaks website also includes a wealth of information for supervisors with topics covering conflict violence prevention plans. TeleLanguage Services: Deer Oaks has the ability to translate any document

  8. Molecular Markers for Biomass Traits: Association, Interaction and Genetic Divergence in Silkworm Bombyx mori.

    PubMed

    Pradeep, Appukuttannair R; Jingade, Anuradha H; Urs, Raje S

    2007-01-01

    Improvement of high yielding, disease resistant silkworm strains became imminent to increase production of silk, which is a major revenue earner for sericulturists. Since environment interacts with phenotype, conventional breeding did not result in commendable yield improvement in synthetic strains of silkworm, Bombyx mori. Identification of DNA markers associated with different economically important biomass traits and its introgression could assist molecular breeding and expression of stabilized high yielding characters, but genetic basis of most quantitative traits in silkworm is poorly understood due to its polygenic control. Correlation analysis (R = 0.9) revealed significant interrelation among biomass traits viz., larval duration (TLD), larval weight (LWT), cocoon weight (CWT), shell weight (SWT), shell ratio (SR) and floss content. PCR using inter simple sequence repeat (ISSR) primers revealed 92% polymorphism among 14 tropical and temperate strains of B. mori, with average diversity index of 0.747. Stepwise multiple regression analysis (MRA) selected 35 ISSR markers positively or negatively correlated with different biomass traits, illustrated polygenic control. ISSR marker 830.8(1050bp) was significantly associated with LWT, CWT, SWT, SR and floss content, indicated its pleiotropic role. Two ISSR markers, 835.5(1950bp) and 825.9(710bp) showed significant association with floss content and TLD. These markers were segregated in F(2) generation and Chi-square test confirmed (chi(2) = ~45; P < 0.05) its genetic contribution to the associated biomass traits. Strains, with both positively and negatively correlated markers, had intermediate mean value for biomass traits (eg. SWT = 0.17 +/- 0.014 g in GNM and Moria) indicated interaction of loci in natural populations. Low yielding Indian strains grouped together by Hierarchical clustering. Chinese and Japanese strains were distributed in the periphery of ALSCAL matrix indicated convergence of genetic characters in Indian strains. Average genetic distance between Chinese strains and Indian strains (0.193) significantly (P < 0.01) varied from that between Chinese and Japanese strains. Interaction of loci and allelic substitutions induced phenotypic plasticity in temperate B. mori populations on tropic adaptation in India. These outcomes show possibility to combine favorable alleles at different QTL to increase larval, cocoon and shell weight. PMID:19662204

  9. Programmed cell death: Cytochemical and X-ray microanalytical characterization of calcium compartments in neuromuscular junctions during the normal breakdown of the intersegmental muscles in the giant silkmoth Antheraea polyphemus

    Microsoft Academic Search

    Jacques Beaulaton; Universitd Blaise

    1988-01-01

    Summary  Calcium stores were cytochemically demonstrated using a combined oxalate—pyroantimonate method in the neuromuscular junctions of the degenerating intersegmental muscles in the giant silkmothAntheraea polyphemus. The elemental composition of punctate precipitates of the reaction product was determined by electron probe X-ray microanalysis of unstained thin sections by energy-dispersive spectrometry and wavelength-dispersive spectrometry. The wavelength-dispersive spectra collected over terminal axons demonstrate a

  10. Proceedings of the 4th Fire in Eastern Oak Forests Conference 69GTR-NRS-P-102 OAK SAVANNA RESTORATION: OAK RESPONSE TO FIRE AND THINNING

    E-print Network

    Proceedings of the 4th Fire in Eastern Oak Forests Conference 69GTR-NRS-P-102 OAK SAVANNA for restoration of oak savanna, oak woodlands, pine-bluestem woodlands, and pine savanna for application but with 4-year burn regime treatment units. We included two additional thinning treatments, the oak-savanna

  11. Oak Ridge Reservation Annual Site

    E-print Network

    Pennycook, Steve

    Report for 2012 on the World Wide Web http://www.ornl.gov/sci/env_rpt/ Project director Joan Hughes Project coordinator Sharon Thompson Department of Energy project manager and Oak Ridge Office coordinator artists Kathy Jones VJ Ewing, Wendy Hames, Terry Bonine, Cindy Johnson Amy Harkey September 2013 Prepared

  12. Cre-mediated targeted gene activation in the middle silk glands of transgenic silkworms (Bombyx mori).

    PubMed

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

    2013-06-01

    Cre-mediated recombination is widely used to manipulate defined genes spatiotemporally in vivo. The present study evaluated the Cre/loxP system in Bombyx mori by establishing two transgenic lines. One line contained a Cre recombinase gene controlled by a sericin-1 gene (Ser1) promoter. The other line contained a loxP-Stop-loxP-DsRed cassette driven by the same Ser1 promoter. The precise deletion of the Stop fragment was found to be triggered by Cre-mediated site-specific excision, and led to the expression of DsRed fluorescence protein in the middle silk glands of all double-transgenic hybrids. This result was also confirmed by phenotypical analysis. Hence, the current study demonstrated the feasibility of Cre-mediated site-specific recombination in B. mori, and opened a new window for further refining genetic tools in silkworms. PMID:23264031

  13. 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.

  14. An emerging functional natural silk biomaterial from the only domesticated non-mulberry silkworm Samia ricini.

    PubMed

    Pal, Shilpa; Kundu, Joydip; Talukdar, Sarmistha; Thomas, Tintu; Kundu, Subhas C

    2013-08-01

    Mulberry silk fibroin is a widely used biomaterial and recent work on non-mulberry silk fibroin also suggests it may have similar uses. We expect silk fibroin from the only domesticated non-mulberry eri silkworm, Samia ricini, to possess useful properties as a biomaterial. Eri silk gland fibroin is a heterodimeric protein of approximately 450?kDa. Cytocompatibility evaluation with fibroblasts and osteoblast-like cells shows good cell attachment, viability and proliferation. The matrices, which have high thermal stability and good swellability, are also haemocompatible. Eri silk production is cost effective as no agronomic practices are required for their host plant cultivation. This fibroin provide new opportunities as an alternative natural functional biomaterial in various biomedical applications. PMID:23733347

  15. 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

  16. Mechanical properties of regenerated Bombyx mori silk fibers and recombinant silk fibers produced by transgenic silkworms.

    PubMed

    Zhu, Zhenghua; Kikuchi, Yuka; Kojima, Katsura; Tamura, Toshiki; Kuwabara, Nobuo; Nakamura, Takashi; Asakura, Tetsuo

    2010-01-01

    Regenerated silk fibroin fibers from the cocoons of silkworm, Bombyx mori, were prepared with hexafluoro solvents, 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) or hexafluoroacetone-trihydrate (HFA), as dope solvents and methanol as coagulation solvent. The regenerated fiber prepared from the HFIP solution showed slightly larger tensile strength when the draw ratio is 1:3 than that of native silk fiber, but the strength of the regenerated fiber with draw ratio 1:3 from the HFA solution is much lower than that of native silk fiber. This difference in the tensile strength of the regenerated silk fibers between two dope solvents comes from the difference in the long-range orientation of the crystalline region rather than that of short-range structural environment such as the fraction of beta-sheet structure. The increase in the biodegradation was observed for the regenerated silk fiber compared with native silk fiber. Preparations of regenerated silk fibroin fibers containing spider silk sequences were obtained by mixing silk fibroins and silk-like proteins with characteristic sequences from a spider, Naphila clavipes, to produce drag-line silk in E. coli in the fluoro solvents. A small increase in the tensile strength was obtained by adding 5% (w/w) of the silk-like protein to the silk fibroin. The production of silk fibroin fibers with these spider silk sequences was also performed with transgenic silkworms. Small increase in the tensile strength of the fibers was obtained without significant change in the elongation-at-break. PMID:20178693

  17. Transgenesis approaches for functional analysis of peptidergic cells in the silkworm Bombyx mori.

    PubMed

    Daubnerová, Ivana; Roller, Ladislav; Zitnan, Dusan

    2009-05-15

    The domestic silkworm, Bombyx mori represents an insect model of great scientific and economic importance. Besides the establishment of a stable germline transformation using the PiggyBac vector, technically feasible methods for in vivo gene delivery and transient gene expression were developed using viral based vectors, especially Sindbis viruses and baculoviruses. The recombinant baculovirus, Autographa californica multiple nucleopolyhedrovirus (AcMNPV), commonly used for large-scale protein production in permissive cell lines or insects, has been used for foreign gene transfer into specific peptidergic cells of B. mori in vivo. Since targeted gene expression is essential for functional analysis of neuropeptide genes and their receptors, the baculovirus-mediated gene transfer can serve as a reliable approach in reverse genetic studies in the silkworm. We review various strategies employing the baculovirus vector system for transient expression of molecular markers and transcription factors in specific peptidergic cells to investigate their roles in B. mori. We also use this system for functional analysis of neuropeptide signaling in the ecdysis behavioral sequence. Our data indicate that the AcMNPV vector is suitable for efficient delivery of foreign genes and their expression directed into specific peptidergic neurons and endocrine cells of B. mori larvae and pupae. However, some modifications of the vector and steps for optimization are necessary to minimize negative effects of viral infection on the host development. The transient gene expression using the AcMNPV and other virus vectors are promising tools for analysis of molecular mechanisms underlying various neuroendocrine processes during development of B. mori. PMID:19111552

  18. Two Adenine Nucleotide Translocase Paralogues Involved in Cell Proliferation and Spermatogenesis in the Silkworm Bombyx mori

    PubMed Central

    Sugahara, Ryohei; Jouraku, Akiya; Nakakura, Takayo; Kusakabe, Takahiro; Yamamoto, Takenori; Shinohara, Yasuo; Miyoshi, Hideto; Shiotsuki, Takahiro

    2015-01-01

    Mitochondrial adenine nucleotide translocase (ANT) specifically acts in ADP/ATP exchange through the mitochondrial inner membrane. This transporter protein thereby plays a significant role in energy metabolism in eukaryotic cells. Most mammals have four paralogous ANT genes (ANT1-4) and utilize these paralogues in different types of cells. The fourth paralogue of ANT (ANT4) is present only in mammals and reptiles and is exclusively expressed in testicular germ cells where it is required for meiotic progression in the spermatocytes. Here, we report that silkworms harbor two ANT paralogues, the homeostatic paralogue (BmANTI1) and the testis-specific paralogue (BmANTI2). The BmANTI2 protein has an N-terminal extension in which the positions of lysine residues in the amino acid sequence are distributed as in human ANT4. An expression analysis showed that BmANTI2 transcripts were restricted to the testis, suggesting the protein has a role in the progression of spermatogenesis. By contrast, BmANTI1 was expressed in all tissues tested, suggesting it has an important role in homeostasis. We also observed that cultured silkworm cells required BmANTI1 for proliferation. The ANTI1 protein of the lepidopteran Plutella xylostella (PxANTI1), but not those of other insect species (or PxANTI2), restored cell proliferation in BmANTI1-knockdown cells suggesting that ANTI1 has similar energy metabolism functions across the Lepidoptera. Our results suggest that BmANTI2 is evolutionarily divergent from BmANTI1 and has developed a specific role in spermatogenesis similar to that of mammalian ANT4. PMID:25742135

  19. Chondroitinase from baculovirus Bombyx mori nucleopolyhedrovirus and chondroitin sulfate from silkworm Bombyx mori.

    PubMed

    Sugiura, Nobuo; Ikeda, Motoko; Shioiri, Tatsumasa; Yoshimura, Mayumi; Kobayashi, Michihiro; Watanabe, Hideto

    2013-12-01

    Chondroitin sulfate (CS) is a linear polysaccharide composed of repeating disaccharide units of glucuronic acid (GlcUA) and N-acetyl-d-galactosamine (GalNAc) with sulfate groups at various positions. Baculovirus is an insect-pathogenic virus that infects Lepidoptera larvae. Recently, we found that the occlusion-derived virus envelope protein 66 (ODV-E66) from Autographa californica nucleopolyhedrovirus (AcMNPV) exhibits chondroitin (CH)-digesting activity with distinct substrate specificity. Here, we demonstrate that the ODV-E66 protein from Bombyx mori nucleopolyhedrovirus (BmNPV) exhibits 92% homology to the amino acid sequence and 83% of the CH lyase activity of ODV-E66 from AcMNPV. ODV-E66 cleaves glycosyl bonds at nonreducing sides of disaccharide units consisting of nonsulfated and 6-O-sulfated GalNAc residues. We then investigated CS in the silkworm, Bombyx mori, which is the host of BmNPV. CS was present in insect tissues such as the midgut, peritrophic membrane, silk gland and skin. The polysaccharide consisted of a nonsulfated disaccharide unit, mono-sulfated disaccharide at Position 4 of the GalNAc residue and mono-sulfated disaccharide at Position 6 of the GalNAc residue. With regard to immunohistochemical analysis, the staining patterns of the silkworm tissues were different among anti-CS antibodies. Chondroitn sulfate that is digestible by ODV-E66 exists sufficiently in the peritrophic membrane protecting the midgut epithelium from ingested pathogens. Our results suggest that ODV-E66 facilitates the primary infection of the virus by digestion of CS in the peritrophic membrane. PMID:24052236

  20. 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. PMID:11258099

  1. Genome-wide analysis of the WW domain-containing protein genes in silkworm and their expansion in eukaryotes.

    PubMed

    Meng, Gang; Dai, Fangyin; Tong, Xiaoling; Li, Niannian; Ding, Xin; Song, Jiangbo; Lu, Cheng

    2015-06-01

    WW domains are protein modules that mediate protein-protein interactions through recognition of proline-rich peptide motifs and phosphorylated serine/threonine-proline sites. WW domains are found in many different structural and signaling proteins that are involved in a variety of cellular processes. WW domain-containing proteins (WWCPs) and complexes have been implicated in major human diseases including cancer as well as in major signaling cascades such as the Hippo tumor suppressor pathway, making them targets for new diagnostics and therapeutics. There are a number of reports about the WWCPs in different species, but systematic analysis of the WWCP genes and its ligands is still lacking in silkworm and the other organisms. In this study, WWCP genes and PY motif-containing proteins have been identified and analyzed in 56 species including silkworm. Whole-genome screening of B. mori identified thirty-three proteins with thirty-nine WW domains located on thirteen chromosomes. In the 39 silkworm WW domains, 15 domains belong to the Group I WW domain; 14 domains were in Group II/III, 9 domains derived from 8 silkworm WWCPs could not be classified into any group, and Group IV contains only one WW domain. Based on gene annotation, silkworm WWCP genes have functions in multi-biology processes. A detailed list of WWCPs from the other 55 species was sorted in this work. In 14,623 silkworm predicted proteins, nearly 18 % contained PY motif, nearly 30 % contained various motifs totally that could be recognized by WW domains. Gene Ontology and KEGG analysis revealed that dozens of WW domain-binding proteins are involved in Wnt, Hedgehog, Notch, mTOR, EGF and Jak-STAT signaling pathway. Tissue expression patterns of WWCP genes and potential WWCP-binding protein genes on the third day of the fifth instar (L5D3) were examined by microarray analysis. Tissue expression profile analysis found that several WWCP genes and poly-proline or PY motif-containing protein genes took tissue- or gender-dependent expression manner in silkworms. We further analyzed WWCPs and PY motif-containing proteins in representative organisms of invertebrates and vertebrates. The results showed that there are no less than 16 and up to 29 WWCPs in insects, the average is 22. The number of WW domains in insects is no less than 19, and up to 47, the average is 36. In vertebrates, excluding the Hydrobiontes, the number of WWCPs is no less than 34 and up to 49, the average is 43. The number of WW domains in vertebrates is no less than 56 and up to 85, the average is 73. Phylogenetic analysis revealed that most homologous genes of the WWCP subfamily in vertebrates were duplicated during evolution and functions diverged. Nearly 1,000 PY motif-containing protein genes were found in insect genomes and nearly 2,000 genes in vertebrates. The different distributions of WWCP genes and PY motif-containing protein genes in different species revealed a possible positive correlation with organism complexity. In conclusion, this comprehensive bio-information analysis of WWCPs and its binding ligands would provide rich fundamental knowledge and useful information for further exploration of the function of the WW domain-containing proteins not only in silkworm, but also in other species. PMID:25424044

  2. 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). PMID:21554801

  3. A fibroin secretion-deficient silkworm mutant, Nd-s D , provides an efficient system for producing recombinant proteins

    Microsoft Academic Search

    Satoshi Inoue; Toshio Kanda; Morikazu Imamura; Guo-Xing Quan; Katsura Kojima; Hiromitsu Tanaka; Masahiro Tomita; Rika Hino; Katsutoshi Yoshizato; Shigeki Mizuno; Toshiki Tamura

    2005-01-01

    The silkworm Nd-sD mutant is silk fibroin-secretion deficient. In the mutant, a disulfide linkage between the heavy (H) and light (L) chains, which is essential for the intracellular transport and secretion of fibroin, is not formed because of a partial deletion of the L-chain gene. To utilize the inactivity of the mutant L-chain, we investigated the possibility of using the

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

    Microsoft Academic Search

    H.. Kurihara; H. Sezutsu; T. Tamura; K. Yamada

    2007-01-01

    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

  5. Toyama Kametaro and Vernon Kellogg: Silkworm Inheritance Experiments in Japan, Siam, and the United States, 1900–1912

    Microsoft Academic Search

    Lisa Onaga

    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\\u000a entomologist Vernon Kellogg. Toyama’s work, based on research conducted in Japan and Siam, came under international scrutiny\\u000a at a time

  6. Effect of VAM fungi and bacterial biofertilizers on mulberry leaf quality and silkworm cocoon characters under semiarid conditions

    Microsoft Academic Search

    D. M. Ram Rao; J. Kodandaramaiah; M. P. Reddy; R. S. Katiyar

    2007-01-01

    The influence of VAM fungi and bacterial biofertilizer (BBF) with 50% reduction in the recommended dose of (N and P) chemical fertilizers on leaf quality traits of mulberry variety (S-13) and its impact on silkworm (PM × NB4D2) growth and cocoon characters were studied under semi-arid conditions. Four different treatments were imposed i.e., T1: Control (only 100% NPK); T2: VAM

  7. Effect of Salicylic Acid on Commercial Traits of the Bivoltine Crossbreed Races of the Silkworm, Bombyx mori L

    Microsoft Academic Search

    S. C. Kochi; B. B. Kaliwal

    Topical application with 100, 200 and 300 ng\\/ml phytohormone salicyclic acid on commercial traits was analysed in bivoltine CSR2, CSR4 and CSR2XCSR4 crossbreed races of the silkworm, Bombyx mori L. The results showed that there was significant increase in larval weight female cocoon weight, male cocoon shell weight, and hatching percentage with 100 and 200 ng\\/ml, silkgland weight, female cocoon

  8. Simplified purification of soluble histidine-tagged green fluorescent protein from cocoon of transgenic silkworm in metal affinity hydroxyapatite chromatography

    Microsoft Academic Search

    Ken Sugo; Tomohiko Yoshitake; Masahiro Tomita; Shintaro Kobayashi; Yae Kurosawa; Katsumi Kawamura; Tsuneo Okuyama

    2011-01-01

    Histidine-tagged proteins have been purified with immobilized nickel affinity chromatography. This method, however, bears some serious disadvantages, including carcinogenicity and potential leakage of the nickel and the handling and disposal costs. We developed a novel purification process for water-soluble histidine-tagged green fluorescent protein from the cocoons of transgenic silkworms, unlike conventional E. coli expression system which sometimes produced insoluble and

  9. Inheritance and linkage analysis of co-dominant SSR markers on the Z chromosome of the silkworm (Bombyx mori L.).

    PubMed

    Miao, Xue-Xia; Li, Wei-Hua; Li, Mu-Wang; Zhao, Yun-Po; Guo, Xian-Ru; Huang, Yong-Ping

    2008-04-01

    Microsatellites or simple sequence repeats (SSRs) are co-dominant molecular markers. When we used fluorescent SSR markers to construct a linkage map for the female heterogametic silkworm (Bombyx mori, ZW), we found that some loci did not segregate in a Mendelian ratio of 1:1 in a backcross population. These loci segregated in a 3:1 ratio of single bands compared with double bands. Further examination of band patterns indicated that three types of SSR bands were present: two homozygotes and one heterozygote. In the beginning, we considered to discard these markers. By scoring male and female F1 individuals, we confirmed that these loci were located on the Z chromosome. Using the sex-linked visible mutation sch (K05) and its wild-type (C108), we constructed an F1 male backcross (BC1M) mapping population. The combination of sch backcross and SSR data enabled us to map the SSR markers to the Z chromosome. By adjusting input parameters based on these data, we were able to use Mapmaker software to construct a linkage map. This strategy takes advantage of co-dominant markers for positional cloning of genes on the Z chromosome. We localized sch to the Z chromosome relative to six SSR markers and one PCR marker, covering a total of 76.1 cM. The sch mutation is an important sex-linked visible mutation widely used in breeding of commercial silkworms (e.g. male silkworm selection rearing). Localization of the sch gene may prove helpful in cloning the gene and developing strains for marker-assisted selection in silkworm breeding. PMID:18426618

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

    Microsoft Academic Search

    Hirotoshi Endo; Hiromichi Nagasawa; Toshiki Watanabe

    2000-01-01

    The crustacean hyperglycemic hormone (CHH) peptide family includes four types of neuropeptide in decapod and isopod crustaceans, and the ion-transport peptide in orthopteran insects. To identify a new member of this family in Insecta, a PCR-based search for cDNAs encoding CHH-family peptides was carried out in the silkworm Bombyx mori. A cDNA, named BmCHHL (Bombyx moriCHH-like protein), with an open

  11. Hyperproduction of Polyhedrin-IGF II Fusion Protein in Silkworm Larvae Infected with Recombinant Bombyx mori Nuclear Polyhedrosis Virus

    Microsoft Academic Search

    YASUMASA MARUMOTO; YOSHINARI SATO; H. Fujiwara; K. Sakano; Y. Saeki; M. Agata; M. Furusawa; S. Maeda

    1987-01-01

    SUMMARY A gene coding for insulin-like growth factor II (IGF II) was constructed from 16 oligodeoxynucleotides synthesized chemically and cloned into EcoRI-SalI sites of pBR322. In this gene an ATG codon for methionine was introduced for cleavage by CNBr at the beginning of mature IGF II. For expressing foreign genes, a new host- vector system, with Bombyx mori silkworm larvae

  12. Nanoparticle-induced morphological transition of Bombyx mori nucleopolyhedrovirus: a novel method to treat silkworm grasserie disease.

    PubMed

    Das, Sumistha; Bhattacharya, Ankita; Debnath, Nitai; Datta, Alokmay; Goswami, Arunava

    2013-07-01

    Grasserie, a polyorganotrophic disease caused by Bombyx mori nucleopolyhedrovirus (BmNPV), accounts for lethal infection to fifth instar silkworm larvae. It was found that nanoparticle (NP)-induced morphological transformation of BmNPV polyhedra could reduce the infectivity of BmNPV both in cell line and in silkworm larvae. Initially, 11 NPs were screened for evaluation of their nature of interaction with polyhedra surface through scanning electron microscopy. Amongst these NPs, lipophilically coated silica nanoparticle (SNPL), alumina nanoparticles in the hexagonal close-packed ? structure and aspartate capped gold nanoparticle transformed polyhedra were tested for their infectivity in B. mori cell line using cytopathic effect and plaque reduction assay. SNPL was evaluated for its bio-efficacy in fifth instar silkworm larvae. The study of polyhedra morphology as a function of NP concentration showed severe 'roughening' of the polyhedra with replacement of the regular facets by a large number of irregular ones by SNPL, and this caused transition of highly infectious polyhedra into a nearly spherical, non-infectious structure. A moderate polyhedra roughening was observed for alumina NPs, and no roughening was noticed for gold NPs. The morphological changes could be correlated with reduction of virus-induced cytopathic effect and plaque formation, and increased survival rate of SNPL transformed polyhedra infected silkworm larvae to 70.09±6.61% after 96 h. In this group, 61.04±8.03% larvae formed normal cocoons from which moths eclosed, laid eggs and larvae emerged. This study could lead to open up newer pathways for designing nano pharmaceuticals to combat other viral diseases. PMID:23588933

  13. Coordination of wing motion and walking suggests common control of zigzag motor program in a male silkworm moth

    Microsoft Academic Search

    Ryohei Kanzaki

    1998-01-01

    The male silkworm moth, Bombyx mori, exhibits a zigzagging pattern as it walks upwind to pheromones. This species usually does not fly, but obvious wing-beating\\u000a accompanies the pheromone-mediated walking. Males supported by a `sled', after having their legs removed, also moved upwind\\u000a in a pheromone plume along zigzagging tracks, indicating that wing-generated thrust and torque result in locomotory paths\\u000a similar

  14. Oak Leaf Roller and Springtime Defoliation of Live Oak Trees 

    E-print Network

    Drees, Bastiaan M.

    2004-03-26

    will eat all of the newly emerged leaves. Two such caterpillars are the oak leaf roller, Archips semiferana (Walker) (Lepi- doptera: Tortricidae), and an associated species, Sparga- nothis pettitana. These insects occur throughout Texas but are most... is seasonal may provide some comfort. Also, wearing broad- rimmed hats and long-sleeved shirts while out- doors may also help prevent contact with these insects. Damage When a tree is defoliated during the growing season, it becomes stressed and occasionally...

  15. Molecular genetic approach for identifying markers associated with yield traits in the silkworm, Bombyx mori using RFLP-STS primers.

    PubMed

    Mohandas, T P; Sethuraman, B N; Saratchandra, B; Chatterjee, S N

    2004-10-01

    Bombyx mori, the mulberry silkworm, exhibits wide variability in yield and developmental attributes. The genetics of yield expression, shown to be of polygenic nature, is poorly studied in silkworm. To identify markers associated with 10 selected yield traits, multiple regression analysis (MRA) and discriminant function analysis (DFA) were applied on 64 markers generated with eight RFLP-derived sequence-tagged-site (STS) primers on the genomic DNA of 20 silkworm stocks of different origin and diverse yield potential. The analyses led to the identification of ten markers showing significant association with the different yield traits. The markers could classify the stocks according to yield potential, irrespective of their origin and status of diapause. Trait means were significantly different for stocks with and with out the associated marker. The inheritance of a marker G2(1300bp), selected at the first step of MRA for five yield traits was shown to segregate in 1:1 ratio in the F2 progeny from a cross between two divergent stocks. The relevance of the STS primers is discussed in the context of applying multiple regression model for identifying markers associated with yield expression and suitability for molecular breeding work in B. mori for yield improvement. PMID:15609576

  16. Molecular markers (RAPD) associated with growth, yield and origin of the silkworm, Bombyx mori L. in India.

    PubMed

    Chatterjee, S N; Pradeep, A R

    2003-12-01

    To identify the molecular markers associated with growth and yield parameters in silkworm, Bombyx mori, RAPD profiles generated with seven UBC primers for fourteen silkworm stocks, originated from China, Japan, India and Russia, were statistically analyzed. Stepwise Multiple Regression Analysis establishes significant association of 45 markers with larval span, growth indices and four cocoon yield parameters relevant for silk production and t-test attest significance of the association of 89.5(1500 bp) and 54.13(300 bp), respectively with longer larval duration and high cocoon weight. The validity of this selection of markers was further supported with Discriminant Function Analysis (DFA) done on the basis of Mahalanobis D2 statistics. The two indices of yield/growth were also tested with DFA, which helped in identifying a few markers and thereby opened scope of using such marker (e.g. 91.11(900 bp)) for incorporating molecular markers in the breeding program for crop improvement in silkworm. PMID:14964827

  17. Inbreeding Effects on Quantitative Traits in Random Mating and Selected Populations of the Mulberry Silkworm, Bombyx mori

    PubMed Central

    Doreswamy, Jamuna; Gopal, Subramanya

    2012-01-01

    The objective of the present study was to estimate the level of inbreeding coefficient during inbreeding of the pedigree of random mating and selected populations of two distinct races of mulberry silkworm, Bombyx mori (Lepidoptera: Bombycidae), in the silkworm germplasm. The six generation data of the two races, namely multivoltine Pure Mysore and bivoltine NB4D2, were studied for inbreeding depression coefficient using the residual maximum likelihood method, utilizing two statistical models by analyzing six quantitative traits, namely, larval weight, cocoon weight, shell weight, shell ratio, pupation rate, and filament length. The results of the present experiment demonstrated that the inbreeding coefficient was significant in Model 1 for most of the economic traits in the random mating populations of both the races compared to those of selected populations. These results suggest that during stock maintenance, application of rigid selection for increased numbers of generations helps to retain original characteristics of the pure races while reducing the deleterious effects of inbreeding. The significance of inbreeding coefficient is discussed with reference to the inbreeding of silk moths in the silkworm germplasm. PMID:23461728

  18. The silkworm GSTe4 is sensitive to phoxim and protects HEK293 cells against UV-induced cell apoptosis.

    PubMed

    Chen, X Y; Liu, J; Zhang, C D; Li, Y F; Liu, T H; Wang, L; Yu, Q Y; Zhang, Y H; Lu, C; Pan, M H

    2015-08-01

    Glutathione S-transferases (GSTs, EC 2.5.1.18) are a family of super enzymes with multiple functions that play a major role in the detoxification of endogenous and xenobiotic compounds. In our previous study, we have predicted 23 putative cytosolic GSTs in the silkworm genome using bioinformatic methods. In this study, we cloned and studied the insect-specific epsilon-class GST gene GSTe4 from the silkworm, Bombyx mori. The recombinant BmGSTe4 (Bac-BmGSTe4) was overexpressed in SF-9 cell lines, and it was found to have effective GST activity. We also found that the expression of BmGSTe4 was especially down-regulated after the silkworms were fumigated with or ingested phoxim. Moreover, BmGSTe4 protected HEK293 cells against UV-induced cell apoptosis. These results demonstrated that BmGSTe4 has GST activity, is sensitive to phoxim, and plays a role in inhibition of UV-induced cell apoptosis. PMID:25850432

  19. Estimating the potential effects of sudden oak death on oak-dependent birds

    Microsoft Academic Search

    William B. Monahan; Walter D. Koenig

    2006-01-01

    Sudden oak death (SOD), a disease induced by the fungus-like pathogen Phytophthora ramorum, threatens to seriously reduce or eliminate several oak species endemic to the west coast of North America. We investigated how the disappearance of one of these species, coast live oak (Quercus agrifolia), may affect populations of five resident oak-affiliated California birds – acorn woodpecker (Melanerpes formicivorus), Nuttall’s

  20. Expression profiling and regulation of genes related to silkworm posterior silk gland development and fibroin synthesis.

    PubMed

    Li, Jian-ying; Yang, Hui-juan; Lan, Tian-yun; Wei, Hao; Zhang, Hua-rong; Chen, Ming; Fan, Wei; Ma, Ying-ying; Zhong, Bo-xiong

    2011-08-01

    The posterior silk gland (PSG) is the most important suborgan responsible for the synthesis and secretion of silk core fibroin proteins in silkworm. Here, we performed genome-scale expression profiling analysis of silkworm PSG at the fourth molting (M4) and at day 1 (V1), day 3 (V3), day 5 (V5), and wandering stage (W) of the fifth instar by microarray analysis with 22?987 probes. We found that the five genes of silk proteins secreted from PSG including fibroin heavy (H) and light (L) chains, P25, seroin 1, and seroin 2 basically showed obvious up-regulation at V3 which lasted to V5, while slight down-regulation at W. The expression of translation-related genes including ribosomal proteins and translation initiation factors generally remained stable from M4 to V5, whereas it showed clear down-regulation at W. Clustering analysis of the 643 significantly differentially expressed transcripts revealed that 43 of the important genes including seroin 1 and sugar transporter protein had co-expression patterns which were consistent with the rate changes of fibroin synthesis and PSG growth. Pathway analysis disclosed that the genes in different clusters might have co-regulations and direct interactions. These genes were supposed to be involved in the fibroin synthesis and secretion. The differential expression of several hormone-related genes also suggested their functions on the regulation of PSG development and fibroin synthesis. 2D gel-based proteomics and phosphoproteomics profiling revealed that the phosphorylated proteins accounted for no more than one-sixth of the total proteins at each stage, which was much lower than the level in normal eukaryotic cells. Changes in the phosphorylation status and levels of several proteins such as actin-depolymerizing factor 1 and enolase might be deeply involved in fibroin secretion and tissue development. Shotgun proteomic profiling combined with label-free quantification analysis on the PSG at V3, V5, and W revealed that many small heat shock proteins (sHSP) were specially expressed at W, which was substantially consistent with the results from 2-DE analysis, and implied the close correlations of sHSP with the physiological states of PSG at W. A majority of significantly up-regulated proteins at V5 were related to ribosome pathway, which was different from the microarray results, implying that the translation-level regulation of ribosomal proteins might be critical for fibroin synthesis. In contrast, the ubiquitin-proteasome pathway related proteins appeared obviously up-regulated at W, suggesting that the programmed cell death process of PSG cells might be started before cocooning. PMID:21657221

  1. Binding and hydrolysis of radiolabeled pheromone and several analogs by male-specific antennal proteins of the mothAntheraea polyphemus.

    PubMed

    Prestwich, G D; Vogt, R G; Riddiford, L M

    1986-02-01

    Sensory hair proteins from antennae of males of the wild silk moth,Antheraea polyphemus (Lepidoptera, Saturniidae) were incubated with radiolabeled 6E,11Z-hexadecadienyl acetate in the presence of unlabeled pheromone analogs as competitive inhibitors. The two extracellular proteins of importance, a highly active sensillar esterase and an abundant 15,000 mol wt binding protein, interact to degrade labeled pheromone less efficiently in the presence of certain unsaturated acetate analogs of the natural pheromone.Enzymatic hydrolysis of the acetate (or diazoacetate) was also examined for three pheromone analogs: [11,12-(3)H2]-6E,11Z-hexadecadienyl diazoacetate, [11,12-(3)H2]-hexadecyl acetate, and [9,10-(3)H2]-9Z-tetradecenyl acetate. The former two are poor substrates at concentrations over four orders of magnitude. The 9Z-14:Ac, however, is the best alternative substrate for this in vitro pheromone metabolism system. Unlabeled 9Z-14: Ac is also the best competitive inhibitor of the hydrolysis of labeled 6E, 11Z-16: Ac. Whereas the tritiated natural pheromone shows a flat response (ca. 40% conversion) to increasing concentrations from 3 × 10(-9) to 3 × 10(-6) M, tritiated 9Z-14: Ac is degraded more rapidly at higher concentrations. PMID:24306783

  2. Snmp-1, a novel membrane protein of olfactory neurons of the silk moth Antheraea polyphemus with homology to the CD36 family of membrane proteins.

    PubMed

    Rogers, M E; Sun, M; Lerner, M R; Vogt, R G

    1997-06-01

    While olfactory neurons of silk moths are well known for their exquisite sensitivity to sex pheromone odorants, molecular mechanisms underlying this sensitivity are poorly understood. In searching for proteins that might support olfactory mechanisms, we characterized the protein profile of olfactory neuron receptor membranes of the wild silk moth Antheraea polyphemus. We have purified and cloned a prominent 67-kDa protein which we have named Snmp-1 (sensory neuron membrane protein-1). Northern blot analysis suggests that Snmp-1 is uniquely expressed in antennal tissue; in situ hybridization and immunocytochemical analyses show that Snmp-1 is expressed in olfactory neurons and that the protein is localized to the cilia, dendrites, and somata but not the axons. Snmp-1 mRNA expression increases significantly 1-2 days before the end of adult development, coincident with the functional maturation of the olfactory system. Sequence analysis suggests Snmp-1 is homologous with the CD36 protein family, a phylogenetically diverse family of receptor-like membrane proteins. CD36 family proteins are characterized as having two transmembrane domains and interacting with proteinaceous ligands; Snmp-1 is the first member of this family identified in nervous tissue. These findings argue that Snmp-1 has an important role in olfaction; possible roles of Snmp-1 in odorant detection are discussed. PMID:9169446

  3. Morphogenesis of the antenna of the male silkmoth, Antheraea polyphemus. I. The leaf-shaped antenna of the pupa from diapause to apolysis.

    PubMed

    Keil, T A; Steiner, C

    1990-01-01

    The antenna of the male silkmoth Antheraea polyphemus is a featherlike structure consisting of a central stem and ca. 120 side branches, which altogether carry about 70,000 olfactory sensilla. We investigate the development during the pupal phase. At the end of diapause, the antennal rudiment consists of a leaf-shaped, one-layered epidermal sac. It is supplied with oxygen via a central main trachea, which gives off numerous thin side branches. These are segmentally arranged into bundles which run to the periphery of the antennal blade. When the epidermis retracts from the pupal cuticle (apolysis; stage 1), it consists of cells which are morphologically uniform. The epidermal cells form a network of long, irregular basal protrusions (epidermal feet), which crisscross the antennal lumen. During the first day post-apolysis (stage 2), the antennal epidermis differentiates into alternating thick 'sensillogenic' and thin 'non-sensillogenic' areas arranged in stripes which run in parallel to the tracheal bundles. Numerous dark, elongated cells, which might be the sensillar stem cells, are scattered in the sensillogenic epithelium. A number of very early sensilla has been found at the distal edges of the sensillogenic stripes in positions which later will be occupied by sensilla chaetica. The whole antennal blade is enveloped by the transparent ecdysial membrane, consisting of the innermost layers of the pupal cuticle which are detached during apolysis. PMID:18620306

  4. Expression of diverse midgut serine proteinases in the sericigenous Lepidoptera Antheraea assamensis (Helfer) is influenced by choice of host plant species.

    PubMed

    Saikia, M; Singh, Y T; Bhattacharya, A; Mazumdar-Leighton, S

    2011-02-01

    Antheraea assamensis is reared on various species of the Lauraceae family from north-east India for its distinctive cocoon silk. We demonstrate differential expression of digestive trypsin and chymotrypsins in larvae feeding on a primary host, Persea bombycina Kosterm., in comparison to larvae feeding on Litsea monopetala Roxb. using in vitro proteolytic assays, zymogram analyses with proteinase inhibitors, restriction digestion of RNA-PCR amplicons and quantitative real-time PCR (RT-PCR). Eight novel members of the serine proteinase gene family were identified, including an intron-spliced trypsin (AaPb4) and seven putative chymotrypsins (AaPb2, AaPb4, AaPb12, AaLm4, AaLm6, AaLm19 and AaLm29). Midgut transcript levels of the putative trypsin were higher in larvae fed P. bombycina whereas levels of transcripts encoding putative chymotrypsins were higher in larvae reared on L. monopetala. Complex, differential expression of sequence divergent midgut serine proteinases may reflect the ability of lepidopteran larvae to feed on different species of host plants. Possible implications of host plant choice on the digestive physiology of A. assamensis are discussed. PMID:20854480

  5. Inhibitors of eicosanoid biosynthesis influencing the transcripts level of sHSP21.4 gene induced by pathogen infections, in Antheraea pernyi.

    PubMed

    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. 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

  7. Purification and characterization of a 1,3-?-D-glucan recognition protein from Antheraea pernyi larve that is regulated after a specific immune challenge

    PubMed Central

    Jinghai, Zhang; Yuntao, Zhang; Jiaoshu, Lin; Tianyi, Wang; Chunfu, Wu; Rong, Zhang

    2013-01-01

    Pattern recognition receptors are known to participate in the activation of Prophenoloxidase system. In this study, a 1,3-?-D-glucan recognition protein was detected for the first time in Antheraea pernyi larvae (Ap-?GRP). Ap-?GRP was purified to 99.9% homogeneity from the hemolymph using traditional chromatographic methods. Ap-?GRP specifically bind 1,3-?-D-glucan and yeast, but not E. coli or M. luteus. The 1,3-?-D-glucan dependent phenoloxidase (PO) activity of the hemolymph inhibited by anti-Ap-?GRP antibody could be recovered by addition of purified Ap-?GRP. These results demonstrate that Ap-?GRP acts as a biosensor of 1,3-?-Dglucan to trigger the Prophenoloxidase system. A trace mount of 1,3-?-D-glucan or Ap-?GRP alone was unable to trigger the proPO system, but they both did. Ap-?GRP was specifically degraded following the activation of proPO with 1,3-?-Dglucan. These results indicate the variation in the amount of Ap-?GRP after specific immune challenge in A. pernyi hemolymph is an important regulation mechanism to immune response. [BMB Reports 2013; 46(5): 264-269] PMID:23710637

  8. Host plant selection by larvae of the muga silk moth, Antheraea assamensis, and the role of the antenna and maxillary palp.

    PubMed

    Bora, D S; Deka, B; Sen, A

    2013-01-01

    The importance of olfactory senses in food preference in fifth instar larvae of Antheraea assamensis Helfer (Lepidoptera: Saturniidae) was examined by subjecting larvae with only antennae or maxillary palpi after microsurgery to food and odor choice tests. Mean percent consumption, total consumption, and choice indices were used as parameters for drawing conclusions. The foods used were two hosts, two non-hosts, and a neutral medium (water). Both antennae and maxillary palpi were fully competent in preference for host plants, Persea bombycina Kostermans (Laurales: Lauraceae) and Litsea polyantha Juss, over the non-hosts, Litsea grandifolia Teschner and Ziziphus jujuba Miller (Rosales: Rhamnaceae). Both were competent in rejecting the non-hosts, L. grandifolia and Z. jujuba. The odor choice test was carried out using a Y-tube olfactometer and showed similar results to the ingestive tests. The results indicate the necessity of functional integration of a combination of olfactory and gustatory sensilla present in different peripheral organs in food acceptance by A. assamensis larvae. PMID:23909481

  9. Effect of ATG initiation codon context motifs on the efficiency of translation of mRNA derived from exogenous genes in the transgenic silkworm, Bombyx mori.

    PubMed

    Tatematsu, Ken-Ichiro; Uchino, Keiro; Sezutsu, Hideki; Tamura, Toshiki

    2014-01-01

    The context sequence motif surrounding the ATG initiation codon influences mRNA translation efficiency and affects protein production; however, the optimal sequence differs among species. To determine the optimal sequence for production of recombinant proteins in a transgenic silkworm, we compared 14-nucleotide context motifs around the ATG (ATG-context) in 50 silkworm genes and found the following consensus: (A/T)AN(A/T)ATCAAAatgN. We were also able to define the least-common motif: CCN(C/G)CGN(C/T/G)(G/C/T)(T/G)atgC, which served as a negative control. To examine the regulatory role of these motifs in protein expression, we constructed reporter plasmids containing different ATG-context motifs together with either the luciferase gene or an enhanced green fluorescent protein (EGFP) gene. These constructs were then used for comparison of luciferase reporter activity and EGFP production in BmN4 cells in vitro as well as in transgenic silkworms in vivo. We detected 10-fold higher luciferase activity in BmN4 cells transfected with the consensus ATG-context motif construct, compared to the negative control plasmid. ELISA measurements of EGFP translation products with the corresponding constructs in BmN4 cells showed consistently similar results. Interestingly, the translation efficiency of the novel consensus ATG-context motif did not show the highest activity in the transgenic silkworms in vivo, except for the fat body. The highest efficiency in the middle and posterior silk glands was produced by the sericin 1 context. Our results show that the ATG-context motifs differ among silkworm tissues. This result is important for the further improvement of the transgenic silkworm system for the production of recombinant proteins. PMID:25674439

  10. Production of a Chimeric Fibroin Light-chain Polypeptide in a Fibroin Secretion-deficient Naked Pupa Mutant of the Silkworm Bombyx mori

    Microsoft Academic Search

    Kazuyuki Mori; Kazunori Tanaka; Yoshimi Kikuchi; Miho Waga; Shou Waga; Shigeki Mizuno

    1995-01-01

    The allelicNd-sandNd-sDmutations of the silkwormBombyx moriare mapped to the same locus as that of the fibroin light (L)-chain gene (Fib-L). The silkworm carrying the homozygousNd-sorNd-sDmutation secretes less than 0.3% of the normal level of fibroin and produces a thin cocoon (naked pupa). In this study, cDNA sequences of theNd-sandNd-sDL-chains were compared with the cDNA and genomic sequences of the L-chain

  11. Genes encoding small heat shock proteins of the silkworm, Bombyx mori.

    PubMed

    Sakano, Daisuke; Li, Bin; Xia, Qingyou; Yamamoto, Kohji; Banno, Yutaka; Fujii, Hiroshi; Aso, Yoichi

    2006-10-01

    Small heat shock protein (sHSP) is a family of ubiquitous polypeptides involved in a variety of physiological phenomena. From the silkworm, Bombyx mori, we isolated and sequenced the following cDNAs encoding sHSPs: shsp19.9, shsp20.1, shsp20.4, shsp20.8, shsp21.4, and shsp23.7. shsp21.4 was nearly twice as large in size as other shsps. The deduced amino acid sequence of sHSP21.4 was similar to that of Drosophila melanogaster CG14207-PA. Other sHSPs were highly similar to each other and, in a phylogenetic tree, formed a cluster including Plodia interpunctella alphaCP25. It was speculated that shsp21.4 has at least one intron in genome while other shsps do not. The transcripts of all shsps were subtle, but were constitutively detected in various tissues. Heat shock triggered a substantial increase in the transcripts other than shsp21.4. The B. mori sHSPs are perhaps classified into at least two groups: sHSP21.4 and others. PMID:17031055

  12. 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. PMID:25302556

  13. Comparative proteome analysis of multi-layer cocoon of the silkworm, Bombyx mori.

    PubMed

    Zhang, Yan; Zhao, Ping; Dong, Zhaoming; Wang, Dandan; Guo, Pengchao; Guo, Xiaomeng; Song, Qianru; Zhang, Weiwei; Xia, Qingyou

    2015-01-01

    Bombyx mori cocoon has a multi-layer structure that provides optimal protection for silkworm pupa. Research on the mechanical properties of the multi-layer structure revealed structure-property relationships of the cocoon. Here, we investigated the protein components of the B. mori cocoon in terms of its multi-layer structure. Liquid chromatography-tandem mass spectrometry identified 286 proteins from the multiple cocoon layers. In addition to fibroins and sericins, we identified abundant protease inhibitors, seroins and proteins of unknown function. By comparing protein abundance across layers, we found that the outermost layer contained more sericin1 and protease inhibitors and the innermost layer had more seroin1. As many as 36 protease inhibitors were identified in cocoons, showing efficient inhibitory activities against a fungal protease. Thus, we propose that more abundant protease inhibitors in the outer cocoon layers may provide better protection for the cocoon. This study increases our understanding of the multi-layer mechanism of cocoons, and helps clarify the biological characteristics of cocoons. The data have been deposited to the ProteomeXchange with identifier PXD001469. PMID:25860555

  14. Proton spin-lattice relaxation in silkworm cocoons: physisorbed water and serine side-chain motions.

    PubMed

    Geppi, Marco; Mollica, Giulia; Borsacchi, Silvia; Cappellozza, Silvia

    2010-03-01

    The molecular dynamic behavior of silkworm cocoons produced by a single Bombyx mori strain was investigated by means of high- and low-resolution solid-state NMR experiments. Cocoons with different moisture content were prepared to study the effects of physisorbed water on their molecular dynamics in the MHz regime, which was probed through the measurement of (1)H T(1) relaxation times at 25 MHz in the 25-95 degrees C temperature range. The water content of the different samples was determined from the analysis of (1)H free-induction decays. In addition to the rotation of methyl groups, mostly from alanine, and to the reorientation of physisorbed water molecules, already identified in previous works as relaxation sinks, the reorientation of serine side-chains was here found to contribute to (1)H T(1) above room temperature. The analysis of the trends of (1)H T(1) versus temperature was carried out in terms of semiempirical models describing the three main motional processes, and indicated that methyl rotation, water reorientation and serine side-chain motions are the most efficient relaxation mechanisms below 0 degrees C, between 0 and 60 degrees C, and above 60 degrees C, respectively. The activation energies were found to decrease passing from serine to water to methyl motions. PMID:20136080

  15. Virulence of an exotoxin A-deficient strain of Pseudomonas aeruginosa toward the silkworm, Bombyx mori.

    PubMed

    Chieda, Yuuka; Iiyama, Kazuhiro; Lee, Jae Man; Kusakabe, Takahiro; Yasunaga-Aoki, Chisa; Shimizu, Susumu

    2011-12-01

    We studied the contribution of exotoxin A to the virulence of Pseudomonas aeruginosa against the silkworm, Bombyx mori. First, an exotoxin A-deficient mutant strain (PAO1toxA) was created, and its virulence compared with that of the parental PAO1 strain. In a short-term mortality assay, the mutant harboring pBBR1MCS2 did not kill B. mori until 120 h after inoculation and complementation of the corresponding gene in trans restored the strain's virulence. Next, to ascertain whether or not it lost all virulence, PAO1toxA (pBBR1MCS2, pGFP) was used in a long-term mortality assay. B. mori inoculated with the mutant strain did not die until early in the 5th instar (240 h after inoculation). However, 50% of the inoculated B. mori died late in the 5th instar or in the early pupal stage (408 h after inoculation). All had died by the pupal stage (600 h after inoculation). The mutant strain was isolated from dead larvae and cocoons. The bacterial population of PAO1toxA in hemolymph reached 4.77 × 10(7) cfu/ml. These results indicated that exotoxin A acts as a virulence factor in B. mori and that other virulence factor(s) are involved during the late stages of infection. PMID:21945328

  16. Comparison of phenotypic and genetic performance of local silkworm groups and two commercial lines.

    PubMed

    Mirhosseini, Seyed Ziaeddin; Nematollahian, Shahla; Ghanipoor, Mani; Seidavi, Alireza

    2010-01-01

    Five Iranian native silkworm groups: Baghdad, Khorasan Orange, Guilan Orange, Khorasan Pink, Khorasan Lemon, and 107 and 110 commercial lines (12 families from each breed) were randomly selected and reared during 2003-2005 (five generations in spring and autumn). In each family, 30 male and 30 female cocoons were individually recorded for weight, shell weight and shell ratio. From among the native groups, the highest average in all three traits belonged to Baghdad and Khorasan Pink, and the lowest to Khorasan Orange and Khorasan Lemon. From among the commercial lines, the highest average in all three traits belonged to 107. In comparing heritability for cocoon weight in native groups, the highest heritability belonged to Guilan Orange (0.5147) and Khorasan Orange (0.5036) and the lowest heritability belonged to Khorasan Pink (0.0967). In the two other traits, the highest heritability belonged to Khorasan Orange and Baghdad and the lowest to Khorasan Pink. In the commercial lines, line110 had higher heritability than line107 for cocoon weight and cocoon shell weight. In all the groups, genetic correlations between cocoon weight and cocoon shell weight were high, expect for the Baghdad group. There was medium or low genetic correlation among cocoon weight, cocoon shell weight and cocoon shell ratio. PMID:21526267

  17. A new continuous cell line from larval ovaries of silkworm, Bombyx mori.

    PubMed

    Khurad, Arun M; Zhang, Min-Juan; Deshmukh, Chanchal G; Bahekar, Ravindra S; Tiple, Ashish D; Zhang, Chuan-Xi

    2009-09-01

    A new continuous cell line from ovarian tissue of commercial variety "Kolar Gold" of silkworm, Bombyx mori, was established and designated as DZNU-Bm-12. The tissue was grown in MGM-448 insect cell culture medium supplemented with 10% fetal bovine serum (FBS) and 3% heat-inactivated B. mori hemolymph at 25 +/- 1 degrees C. The migration of partially attached small round refractive cells from the fragments of ovarioles began from the beginning of explantation. The cells multiplied partially attached in the primary culture initially, and some of them become freely suspended after 20 passages. The cells were adapted to MGM-448 and TNM-FH media each with 10% FBS and the population doubling time of cell line was about 36 and 24 hr, respectively. The chromosome number was near diploid at initial passages and slightly increased at 176th passage, but a few tetraploids and hexaploids were also observed. DNA profiles using simple sequence repeat loci established the differences between DZNU-Bm-12 and DZNU-Bm-1 and most widely used Bm-5 and BmN cell lines. The cell line was found susceptible to B. mori nucleopolyhedrovirus (BmNPV) with 85-90% of the cells harboring BmNPV and having an average of 3-17 OBs/infected cell. We suggest the usefulness of this cell line in BmNPV-based baculoviral expression system and also for studying in vitro virus replication. PMID:19357932

  18. 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. PMID:26108630

  19. Transgenic protein production in silkworm silk glands requires cathepsin and chitinase of Autographa californica multicapsid nucleopolyhedrovirus.

    PubMed

    Wöltje, Michael; Böbel, Melanie; Rheinnecker, Michael; Tettamanti, Gianluca; Franzetti, Eleonora; Saviane, Alessio; Cappellozza, Silvia

    2014-05-01

    The silkworm Bombyx mori represents an established in vivo system for the production of recombinant proteins. Baculoviruses have been extensively investigated and optimised for the expression of high protein levels inside the haemolymph of larvae and pupae of this lepidopteran insect. Current technology includes deletion of genes responsible for the activity of virus-borne proteases, which in wild-type viruses, cause liquefaction of the host insect and enhance horizontal transmission of newly synthesised virus particles. Besides the haemolymph, the silk gland of B. mori provides an additional expression system for recombinant proteins. In this paper, we investigated how silk gland can be efficiently infected by a Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV). We demonstrated that the viral chitinase and the cysteine protease cathepsin are necessary to permit viral entry into the silk gland cells of intrahaemocoelically infected B. mori larvae. Moreover, for the first time, we showed AcMNPV crossing the basal lamina of silk glands in B. mori larvae, and we assessed a new path of infection of silk gland cells that can be exploited for protein production. PMID:24477386

  20. Wound healing properties of a 3-D scaffold comprising soluble silkworm gland hydrolysate and human collagen.

    PubMed

    Kim, Kyu-Oh; Lee, Youngjun; Hwang, Jung-Wook; Kim, Hojin; Kim, Sun Mi; Chang, Sung Woon; Lee, Heui Sam; Choi, Yong-Soo

    2014-04-01

    Biomaterials that serve as scaffolds for cell proliferation and differentiation are increasingly being used in wound repair. In this study, the potential regenerative properties of a 3-D scaffold containing soluble silkworm gland hydrolysate (SSGH) and human collagen were evaluated. The scaffold was generated by solid-liquid phase separation and a freeze-drying method using a homogeneous aqueous solution. The porosity, swelling behavior, protein release, cytotoxicity, and antioxidative properties of scaffolds containing various ratios of SSGH and collagen were evaluated. SSGH/collagen scaffolds had a high porosity of 61-81% and swelling behavior studies demonstrated a 50-75% increase in swelling, along with complete protein release in the presence of phosphate-buffered saline. Cytocompatibility of the SSGH/collagen scaffold was demonstrated using mesenchymal stem cells from human umbilical cord. Furthermore, SSGH/collagen efficiently attenuated oxidative stress-induced cell damage. In an in vivo mouse model of wound healing, the SSGH/collagen scaffold accelerated wound re-epithelialization over a 15-day period. Overall, the microporous SSGH/collagen 3-D scaffold maintained optimal hydration of the exposed tissues and decreased wound healing time. These results contribute to the generation of advanced wound healing materials and may have future therapeutic implications. PMID:24503353

  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. PMID:25808998

  2. 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. PMID:25958343

  3. Structure-function relationship of SW-AT-1, a serpin-type protease inhibitor in silkworm.

    PubMed

    Liu, Cheng; Han, Yue; Chen, Xi; Zhang, Wei

    2014-01-01

    Although SW-AT-1, a serpin-type trypsin inhibitor from silkworm (Bombyx mori), was identified in previous study, its structure-function relationship has not been studied. In this study, SW-AT-1 was cloned from the body wall of silkworm and expressed in E. coli. rSW-AT-1 inhibited both trypsin and chymotrypsin in a concentration-dependent manner. The association rate constant for rSW-AT-1 and trypsin is 1.31×10-5 M-1s-1, for rSW-AT-1 and chymotrpsin is 2.85×10-6 M-1s-1. Circular dichroism (CD) assay showed 33% ?-helices, 16% ?-sheets, 17% turns, and 31% random coils in the secondary structure of the protein. Enzymatic and CD analysis indicated that rSW-AT-1 was stable at wide pH range between 4-10, and exhibited the highest activity at weakly acidic or alkaline condition. The predicted three-dimensional structure of SW-AT-1 by PyMOL (v1.4) revealed a deductive reactive centre loop (RCL) near the C-terminus, which was extended from the body of the molecule. In addition to trypsin cleavage site in RCL, matrix-assisted laser desorption ionization time of flight mass spectrometry indicated that the chymotrypsin cleavage site of SW-AT-1 was between F336 and T337 in RCL. Directed mutagenesis indicated that both the N- and C-terminal sides of RCL have effects on the activity, and G327 and E329 played an important role in the proper folding of RCL. The physiological role of SW-AT-1 in the defense responses of silkworm were also discussed. PMID:24901510

  4. An optimized sericin-1 expression system for mass-producing recombinant proteins in the middle silk glands of transgenic silkworms.

    PubMed

    Wang, Feng; Xu, Hanfu; Yuan, Lin; Ma, Sanyuan; Wang, Yuancheng; Duan, Xiaoli; Duan, Jianping; Xiang, Zhonghuai; Xia, Qingyou

    2013-10-01

    The middle silk gland (MSG) of silkworm is thought to be a potential host for mass-producing valuable recombinant proteins. Transgenic MSG expression systems based on the usage of promoter of sericin1 gene (sericin-1 expression system) have been established to produce various recombinant proteins in MSG. However, further modifying the activity of the sericin-1 expression system to yield higher amounts of recombinant proteins is still necessary. In this study, we provide an alternative modification strategy to construct an efficient sericin-1 expression system by using the hr3 enhancer (hr3 CQ) from a Chongqing strain of the Bombyx mori nuclear polyhedrosis virus (BmNPV) and the 3'UTRs of the fibroin heavy chain (Fib-HPA), the fibroin light chain (Fib-LPA), and Sericin1 (Ser1PA) genes. We first analyzed the effects of these DNA elements on expression of luciferase, and found that the combination of hr3 CQ and Ser1PA was most effective to increase the activity of luciferase. Then, hr3 CQ and Ser1PA were used to modify the sericin1 expression system. Transgenic silkworms bearing these modified sericin1 expression vectors were generated by a piggyBac transposon mediated genetic transformation method. Our results showed that mRNA level of DsRed reporter gene in transgenic silkworms containing hr3 CQ and Ser1PA significantly increased by 9 fold to approximately 83 % of that of endogenous sericin1. As the results of that, the production of recombinant RFP increased by 16 fold to 9.5 % (w/w) of cocoon shell weight. We conclude that this modified sericin-1 expression system is efficient and will contribute to the MSG as host to mass produce valuable recombinant proteins. PMID:23435751

  5. 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

  6. A 1H-NMR Based Study on Hemolymph Metabolomics in Eri Silkworm after Oral Administration of 1-Deoxynojirimycin.

    PubMed

    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

  7. Molecular cloning and functional characterization of the dual oxidase (BmDuox) gene from the silkworm Bombyx mori.

    PubMed

    Hu, Xiaolong; Yang, Rui; Zhang, Xing; Chen, Lin; Xiang, Xingwei; Gong, Chengliang; Wu, Xiaofeng

    2013-01-01

    Reactive oxygen species (ROS) from nicotinamide adenine dinucleotide phosphate (NADPH) oxidases and their related dual oxidases are known to have significant roles in innate immunity and cell proliferation. In this study, the 5,545 bp cDNA of the silkworm Bombyx mori dual oxidase (BmDuox) gene containing a full-length open reading frame was cloned. It was shown to include an N-terminal signal peptide consisting of 28 amino acid residues, a 240 bp 5'-terminal untranslated region (5'-UTR), an 802 bp 3'-terminal region (3'-UTR), which contains nine ATTTA motifs, and a 4,503 bp open reading frame encoding a polypeptide of 1,500 amino acid residues. Structural analysis indicated that BmDuox contains a typical peroxidase domain at the N-terminus followed by a calcium-binding domain, a ferric-reducing domain, six transmembrane regions and binding domains for flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide (NAD). Transcriptional analysis revealed that BmDuox mRNA was expressed more highly in the head, testis and trachea compared to the midgut, hemocyte, Malpighian tube, ovary, fat bodies and silk glands. BmDuox mRNA was expressed during all the developmental stages of the silkworm. Subcellular localization revealed that BmDoux was present mainly in the periphery of the cells. Some cytoplasmic staining was detected, with rare signals in the nucleus. Expression of BmDuox was induced significantly in the larval midgut upon challenge by Escherichia coli and Bombyx mori nucleopolyhedrovirus (BmNPV). BmDuox-deleted larvae showed a marked increase in microbial proliferation in the midgut after ingestion of fluorescence-labeled bacteria compared to the control. We conclude that reducing BmDuox expression greatly increased the bacterial load, suggesting BmDuox has an important role in inhibiting microbial proliferation and the maintenance of homeostasis in the silkworm midgut. PMID:23936382

  8. Molecular Cloning and Functional Characterization of the Dual Oxidase (BmDuox) Gene from the Silkworm Bombyx mori

    PubMed Central

    Hu, Xiaolong; Yang, Rui; Zhang, Xing; Chen, Lin; Xiang, Xingwei; Gong, Chengliang; Wu, Xiaofeng

    2013-01-01

    Reactive oxygen species (ROS) from nicotinamide adenine dinucleotide phosphate (NADPH) oxidases and their related dual oxidases are known to have significant roles in innate immunity and cell proliferation. In this study, the 5,545 bp cDNA of the silkworm Bombyx mori dual oxidase (BmDuox) gene containing a full-length open reading frame was cloned. It was shown to include an N-terminal signal peptide consisting of 28 amino acid residues, a 240 bp 5?-terminal untranslated region (5?-UTR), an 802 bp 3?-terminal region (3?-UTR), which contains nine ATTTA motifs, and a 4,503 bp open reading frame encoding a polypeptide of 1,500 amino acid residues. Structural analysis indicated that BmDuox contains a typical peroxidase domain at the N-terminus followed by a calcium-binding domain, a ferric-reducing domain, six transmembrane regions and binding domains for flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide (NAD). Transcriptional analysis revealed that BmDuox mRNA was expressed more highly in the head, testis and trachea compared to the midgut, hemocyte, Malpighian tube, ovary, fat bodies and silk glands. BmDuox mRNA was expressed during all the developmental stages of the silkworm. Subcellular localization revealed that BmDoux was present mainly in the periphery of the cells. Some cytoplasmic staining was detected, with rare signals in the nucleus. Expression of BmDuox was induced significantly in the larval midgut upon challenge by Escherichia coli and Bombyx mori nucleopolyhedrovirus (BmNPV). BmDuox-deleted larvae showed a marked increase in microbial proliferation in the midgut after ingestion of fluorescence-labeled bacteria compared to the control. We conclude that reducing BmDuox expression greatly increased the bacterial load, suggesting BmDuox has an important role in inhibiting microbial proliferation and the maintenance of homeostasis in the silkworm midgut. PMID:23936382

  9. pH induced changes in the rheology of silk fibroin solution from the middle division of Bombyx mori silkworm.

    PubMed

    Terry, Ann E; Knight, David P; Porter, David; Vollrath, Fritz

    2004-01-01

    The rheological properties of fibroin silk solutions extracted from the middle division of Bombyx mori silkworms were examined. Acidification of the solutions with acetic acid vapor gelled the material, a process which at short time scales could be reversed by exposure to ammonia vapor. The solution could also be converted to sol from the gel state by the addition of EDTA. The possible mechanisms for gel formation in fibroin solutions is discussed as are the implications for the process of spinning silk fibers. PMID:15132659

  10. Sericin, a protein derived from silkworms, accelerates the proliferation of several mammalian cell lines including a hybridoma

    Microsoft Academic Search

    Satoshi Terada; Taeko Nishimura; Masahiro Sasaki; Hideyuki Yamada; Masao Miki

    2002-01-01

    Sericin, a constituent of the silkworm cocoon, was added to the culture of four mammalian cell lines: murine hybridoma 2E3-O,human\\u000a hepatoblastoma HepG2, human epithelial HeLa and human embryonal kidney 293 cells. The proliferation of all cell lineswas accelerated\\u000a in the presence of sericin. The hybridoma cellline was further studied. The 2E3-O cell line was so well adapted to serum-free\\u000a medium

  11. DNA fingerprinting using AFLP markers to search for markers associated with yield attributes in the silkworm, Bombyx mori.

    PubMed

    Gaviria, Duverney A; Aguilar, Enrique; Serrano, Herman J; Alegria, Alvaro H

    2006-01-01

    This study was carried out on 11 Chinese and 12 Japanese silkworm strains maintained by the Center for the Technological Development of Sericulture (CDTS) germplasm bank, located in Pereira, Colombia. The goals were to determine the genetic population structure of the two groups and the association between molecular markers (AFLPs) and important productivity characters. Group analysis showed the separation of the strains according to their geographic origin. The molecular markers and the productivity characters were correlated by multiple variance analysis. The analysis permitted the identification of molecular markers associated with the cocoon weight or the shell weight separately. Some markers were associated with both characters. PMID:19537986

  12. Oak Leaf Blister Theresa Badurek, Urban Horticulture Extension Agent

    E-print Network

    Jawitz, James W.

    Oak Leaf Blister Theresa Badurek, Urban Horticulture Extension Agent It's that time of year again spring! Wait a minute oak leaves blistering? Yes, this is a common sight in the spring. Oak leaf blister (Taphrina caerulescens) is a common fungal leaf disease on oaks in Florida. It can affect any

  13. Morphogenesis of the antenna of the male silkmoth, Antheraea polyphemus. II. Differential mitoses of 'dark' precursor cells create the Anlagen of sensilla.

    PubMed

    Keil, T A; Steiner, C

    1990-01-01

    The antenna of the male silkmoth Antheraea polyphemus develops from a one-layered, flattened epidermal sac during the pupal phase. Within the first day post-apolysis (developmental stages 1 and 2), this epithelium differentiates into 'sensillogenic' and 'nonsensillogenic' regions, while numerous slender 'dark cells' interpreted as the precursor cells of sensilla arise in the former. Approximately between the first and second day post-apolysis (developmental stage 3), the dark cells retract to the apical pole of the epidermis, assume a round shape, and undergo a series of differential mitoses with spindles usually oriented parallel to the epidermal surface. These mitoses finally yield the Anlagen of the olfactory sensilla trichodea, each consisting of mostly 6-7 dark cells arranged side by side. In most of the Anlagen, 3-4 of these cells are situated more basally, each giving off a slender apical process which together are arranged in a fascicle. These are the prospective 2-3 sensory neurons plus the thecogen cell, which most probably is a sister cell of the former. Three additional cells are arranged more apically and partly enclose the fascicle of presumed sensory and thecogen cell processes. These are interpreted as the trichogen plus 2 tormogen cells, one of the latter degenerating later during development. In the basal region of the sensillogenic epidermis, massive signs of cell degeneration have been found. At stage 3, the basal epidermal feet in the non-sensillogenic regions have assumed a more uniform orientation as compared with the preceding stages. PMID:18620326

  14. Ligand binding turns moth pheromone-binding protein into a pH sensor: effect on the Antheraea polyphemus PBP1 conformation.

    PubMed

    Katre, Uma V; Mazumder, Suman; Prusti, Rabi K; Mohanty, Smita

    2009-11-13

    In moths, pheromone-binding proteins (PBPs) are responsible for the transport of the hydrophobic pheromones to the membrane-bound receptors across the aqueous sensillar lymph. We report here that recombinant Antheraea polyphemus PBP1 (ApolPBP1) picks up hydrophobic molecule(s) endogenous to the Escherichia coli expression host that keeps the protein in the "open" (bound) conformation at high pH but switches to the "closed" (free) conformation at low pH. This finding has bearing on the solution structures of undelipidated lepidopteran moth PBPs determined thus far. Picking up a hydrophobic molecule from the host expression system could be a common feature for lipid-binding proteins. Thus, delipidation is critical for bacterially expressed lipid-binding proteins. We have shown for the first time that the delipidated ApolPBP1 exists primarily in the closed form at all pH levels. Thus, current views on the pH-induced conformational switch of PBPs hold true only for the ligand-bound open conformation of the protein. Binding of various ligands to delipidated ApolPBP1 studied by solution NMR revealed that the protein in the closed conformation switches to the open conformation only at or above pH 6.0 with a protein to ligand stoichiometry of approximately 1:1. Mutation of His(70) and His(95) to alanine drives the equilibrium toward the open conformation even at low pH for the ligand-bound protein by eliminating the histidine-dependent pH-induced conformational switch. Thus, the delipidated double mutant can bind ligand even at low pH in contrast to the wild type protein as revealed by fluorescence competitive displacement assay using 1-aminoanthracene and solution NMR. PMID:19758993

  15. Structural basis of ligand binding and release in insect pheromone-binding proteins: NMR structure of Antheraea polyphemus PBP1 at pH 4.5.

    PubMed

    Damberger, Fred F; Ishida, Yuko; Leal, Walter S; Wüthrich, Kurt

    2007-11-01

    The NMR structure of the Antheraea polyphemus pheromone-binding protein 1 at pH 4.5, ApolPBP1A, was determined at 20 degrees C. The structure consists of six alpha-helices, which are arranged in a globular fold that encapsulates a central helix alpha7 formed by the C-terminal polypeptide segment 131-142. The 3D arrangement of these helices is anchored by the three disulfide bonds 19-54, 50-108 and 97-117, which were identified by NMR. Superposition of the ApolPBP1A structure with the structure of the homologous pheromone-binding protein of Bombyx mori at pH 4.5, BmorPBPA, yielded an rmsd of 1.7 A calculated for the backbone heavy-atoms N, Calpha and C' of residues 10-142. In contrast, the present ApolPBP1A structure is different from a recently proposed molecular model for a low-pH form of ApolPBP1 that does not contain the central helix alpha7. ApolPBP1 exhibits a pH-dependent transition between two different globular conformations in slow exchange on the NMR chemical shift timescale similar to BmorPBP, suggesting that the two proteins use the same mechanism of ligand binding and ejection. The extensive sequence homology observed for pheromone-binding proteins from moth species further implies that the previously proposed mechanism of ligand ejection involving the insertion of a C-terminal helix into the pheromone-binding site is a general feature of pheromone signaling in moths. PMID:17884092

  16. Ultrastructural studies on neuromuscular contacts and the formation of junctions in the flight muscle of Antheraea polyphemus (Lep.). II. Changes after motor nerve section.

    PubMed

    Nüesch, H; Stocker, R F

    1975-12-10

    In the moth Antheraea polyphemed at the onset of adult development. The subsequent breakdown of the isolated motor stulongated vesicles similar in structure to channels of smooth ER, appear in large numbers in the axoplasm. Their nature as well as the functional aspects of early axonal changes are discussed. From the 7th day onward two types of axonal breakdown become prominent. The first is characterized 0y swelling axon profiles, distorted vesicles and strongly shrunken mitochondria, uhile shrinking axon profiles containing tightly packed mitochondria and unaltered vesicles are typical of the second. Both types presumably take place independently of each other in different axon terminals. Axons and the contents of at least the first type are finally removed by transformation into lamellar bodies. Glial processes obviously behave independently of degenerating terminals; they loose any contact with them and never act as phagocytes for axon remnants. During the whole period of breakdown undifferentiated contacts between nerve fibers and muscle anlagen are present but synaptic structures as in normal developing dlm have never been observed. This fact, in comparison with earlier studies, suggests a lack of trophic nervous activity on the muscle anlagen tissue. A short time after removal of the isolated stumps new nerve tracts appear between dlm-fibers (which are, of course, strongly retarded in development). They are presumably sensory wing nerves which lack a guide structure to the central target, due to axotomy. Neuromuscular contacts or even junctions formed by axons of these nerves have occasionally been detected on the dlm. Their nature is discussed. Wallerian axon degeneration is compared to the normal, metamorphic breakdown of the innervation of the larval dlm-precursor. In contrast to the former, glial processes here remain in contact with the terminals. Glia and axons first swell. Then most glial processes are transformed into lamellar bodies whereas neurites shrink and become electron-dense. Axonal organelles remain intact for a long period. PMID:1201608

  17. Ultrastructural studies on neuromuscular contacts and the formation of junctions in the flight muscle of Antheraea polyphemus (Lep.) I. Normal adult development.

    PubMed

    Stocker, R F; Nüesch, H

    1975-06-01

    The ultrastructure of neuromuscular connections on developing dorsolongitudinal flight muscles were studied in the moth Antheraea polyphemus. Undifferentiated membrane contacts between axon terminals and muscle-fiber anlagen are present in the diapause pupa. They persist during the period of nerve outgrowth, which probably provides a pathway of contact guidance. By the 4th day of adult development some of these contact areas have differentiated into structures similar to neuromuscular junctions although differentiation of muscle structure does not start earlier than the eighth day. Dense-cored vesicles are abundant in many axon terminals at the beginning of development. They later decrease in number quite rapidly. The significance of the above-mentioned early junctions, their possible mode of action and the role of the dense-cored vesicles are discussed. It is proposed that they exercise a stimulating (trophic) influence on the growth of the undifferentiated muscular tissue. The imaginal neuromuscular junctions are formed during the second half of adult development. Clusters of vesicles and electron-dense depositions along the inner face of the axo- and lemma seem to initiate junction formation. Glial processes then grow between the axo- and sarcolemma and divide the large contact area into several small segments. Mutual invaginations and protrusions of the sarcolemma and the glial cell membrane subsequently form an extensive "rete synapticum." Six days before eclosion the glial and sarcoplasmic parts of the rete synapticum are similar in size. Up to eclosion, all glial processes shrink and increase in electron density. Most of the observations are discussed also in relation to findings in vertebrates. PMID:1149098

  18. Antheraea pernyi Silk Fibroin-Coated PEI/DNA Complexes for Targeted Gene Delivery in HEK 293 and HCT 116 Cells

    PubMed Central

    Liu, Yu; You, Renchuan; Liu, Guiyang; Li, Xiufang; Sheng, Weihua; Yang, Jicheng; Li, Mingzhong

    2014-01-01

    Polyethylenimine (PEI) has attracted much attention as a DNA condenser, but its toxicity and non-specific targeting limit its potential. To overcome these limitations, Antheraea pernyi silk fibroin (ASF), a natural protein rich in arginyl-glycyl-aspartic acid (RGD) peptides that contains negative surface charges in a neutral aqueous solution, was used to coat PEI/DNA complexes to form ASF/PEI/DNA ternary complexes. Coating these complexes with ASF caused fewer surface charges and greater size compared with the PEI/DNA complexes alone. In vitro transfection studies revealed that incorporation of ASF led to greater transfection efficiencies in both HEK (human embryonic kidney) 293 and HCT (human colorectal carcinoma) 116 cells, albeit with less electrostatic binding affinity for the cells. Moreover, the transfection efficiency in the HCT 116 cells was higher than that in the HEK 293 cells under the same conditions, which may be due to the target bonding affinity of the RGD peptides in ASF for integrins on the HCT 116 cell surface. This result indicated that the RGD binding affinity in ASF for integrins can enhance the specific targeting affinity to compensate for the reduction in electrostatic binding between ASF-coated PEI carriers and cells. Cell viability measurements showed higher cell viability after transfection of ASF/PEI/DNA ternary complexes than after transfection of PEI/DNA binary complexes alone. Lactate dehydrogenase (LDH) release studies further confirmed the improvement in the targeting effect of ASF/PEI/DNA ternary complexes to cells. These results suggest that ASF-coated PEI is a preferred transfection reagent and useful for improving both the transfection efficiency and cell viability of PEI-based nonviral vectors. PMID:24776757

  19. Attachment and Growth of Fibroblast Cells on Silk Fibroin

    Microsoft Academic Search

    N. Minoura; S. I. Aiba; M. Higuchi; Y. Gotoh; M. Tsukada; Y. Imai

    1995-01-01

    The attachment and growth of fibroblast cells (L-929) on matrices of silk fibroin from Bombyx mori domestic silkworm (DSF) and Antheraea pernyi wild silkworm (WSF) were studied by a cell culture method. The performance of the two kinds of silk fibroin was compared to that of collagen. DSF exhibited as high a cell attachment and growth as collagen did. The

  20. Novel enhancer and promoter elements indispensable for the tissue-specific expression of the sericin-1 gene of the silkworm Bombyx mori

    Microsoft Academic Search

    Shigeharu Takiya; Hiroshige Inoue; Mai Kimoto

    2011-01-01

    Sericins are glue proteins produced specifically in the middle silk gland (MSG) of the silkworm Bombyx mori, while the silk fiber protein, fibroin, is produced in the posterior silk gland (PSG). These silk proteins are expected to be useful biomaterials in medical technology as well as biotechnology. In this study, we analyzed promoter elements of the sericin-1 gene (ser1) in

  1. The silk cocoon of the silkworm, Bombyx mori: Macro structure and its influence on transmural diffusion of oxygen and water vapor

    Microsoft Academic Search

    Bonnie Blossman-Myer; Warren W. Burggren

    2010-01-01

    The cocoon of insect larvae is thought to help conserve water while affording mechanical protection. If the cocoon is a barrier to water loss, then it must also impose a barrier to inward oxygen diffusion. We tested this hypothesis in pupae of the silkworm, Bombyx mori. The rate of water loss and oxygen uptake (V?O2) at 25°C was measured in

  2. [Molecular mapping of test mapping strain for 18th linkage group recessive genes elp, ch-2 and mln in silkworm (Bombyx mori)].

    PubMed

    Liu, Xian-Fang; Ma, Xiao; Hou, Cheng-Xiang; Li, Bing; Li, Mu-Wang

    2013-03-01

    The ellipsoid egg, the second recessive gene of chocolate larvae, and melanism are controlled by three recessive genes, elp, ch-2, and mln in silkworm, respectively. Their order and genetic distance have been scheduled in established linkage group. Owing to lack of crossing over in females, the reciprocal backcrossed F1(BC1) progenies were bred for linkage analysis using the wild type silkworm strain P50(+elp+ch-2+mln /+elp+ch-2+mln) and W18 with ellipsoid egg, the second recessive gene of chocolate larvae, and melanism (elp ch-2 mln / elp ch-2 mln). In this research, we mapped three mutant genes, elp, ch-2, and mln on the chromosome 18 based on the SSR linkage map and STS markers designed based on silkworm genome sequence. The established linkage group, molecular linkage group, and the physic map of chromosome 18 had been corresponded. The genetic distance for this chromosome in this research was 94.2 cM, and the order of the mutants and molecular markers were consistent with the established silkworm linkage maps and the fine genome sequences. This research will lay important bases for map-based cloning for other mutants on chromosome 18. PMID:23575544

  3. Electron transport and bulk-like behavior of Wiedemann-Franz law for sub-7 nm-thin iridium films on silkworm silk.

    PubMed

    Lin, Huan; Xu, Shen; Zhang, Yu-Qing; Wang, Xinwei

    2014-07-23

    For ultrathin metallic films, either supported or free-standing, the inside nanocrystalline nature significantly reduces the electron and thermal transport. Quantum mechanical reflection of electrons at the grain boundary reduces the electrical conductivity further than the thermal conductivity, leading to a Lorenz number in the order of 7.0 × 10(-8) W ? K(-2), much higher than that of the bulk counterpart. We report on a finding that for ultrathin (0.6-6.3 nm) iridium films coated on degummed silkworm silk fibroin, the electron transport is around 100-200% higher than that of the same film on glass fiber, even though the grain size of Ir film on silkworm silk is smaller than that on glass fiber. At the same time, the thermal conductivity of the Ir film is smaller or close to that of the film on glass fiber. Its Lorenz number is found close to that of bulk crystalline Ir despite the nanocrystalline structure in the Ir films. This is similar to the behavior of metallic glasses. Our study of gold films on silkworm silk reveals the same trend of change as compared to that on glass fiber. Electron hopping and tunneling in silkworm silk is speculated to be responsible for the observed electron transport. The finding points out that silk could provide a better substrate for flexible electronics with significantly faster electron transport. PMID:24988039

  4. 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

    Microsoft Academic Search

    Y. B. Gaikwad; S. M. Gaikwad; G. P. Bhawane

    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

  5. NOT ALL OAK GALL WASPS GALL OAKS: THE DESCRIPTION OF DRYOCOSMUS RILEYPOKEI, A NEW, APOSTATE SPECIES OF CYNIPINI

    E-print Network

    Hammerton, James

    NOT ALL OAK GALL WASPS GALL OAKS: THE DESCRIPTION OF DRYOCOSMUS RILEYPOKEI, A NEW, APOSTATE SPECIES-mail: simorita@ncsu.edu) Abstract.--Cynipini gall wasps (Hymenoptera: Cynipidae) are commonly known as oak gall wasps for their almost exclusive use of oak (Quercus spp.; Fagaceae) as their host plant. Previously

  6. Sudden Oak Death Blitz: Native oaks need our help APRIL 19, 2013 6:57 PM BILL PRAMUK

    E-print Network

    California at Berkeley, University of

    Sudden Oak Death Blitz: Native oaks need our help APRIL 19, 2013 6:57 PM · BILL PRAMUK On a recent ominous than the invasive exotic plants: a dying coast live oak. The walk was organized by the Napa laurels were scattered here and there along with the native oaks. Down the slope, that dying coast live

  7. 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.

  8. Production of a recombinant mouse monoclonal antibody in transgenic silkworm cocoons.

    PubMed

    Iizuka, Masashi; Ogawa, Shingo; Takeuchi, Atsushi; Nakakita, Shinichi; Kubo, Yuhki; Miyawaki, Yoshitaka; Hirabayashi, Jun; Tomita, Masahiro

    2009-10-01

    In the present study, we describe the production of transgenic silkworms expressing a recombinant mouse mAb in their cocoons. Two transgenic lines, L- and H-, were generated that carried cDNAs encoding the L- and H-chains of a mouse IgG mAb, respectively, under the control of the enhancer-linked sericin-1 promoter. Cocoon protein analysis indicated that the IgG L- or H-chain was secreted into the cocoons of each line. We also produced a transgenic line designated L/H, which carried both cDNAs, by crossing the L- and H-lines. This line efficiently produced the recombinant mAb as a fully assembled H(2)L(2) tetramer in its cocoons, with negligible L- or H-chain monomer and H-chain dimer production. Thus, the H(2)L(2) tetramer was synthesized in, and secreted from, the middle silk gland cells. Crossing of the L/H-line with a transgenic line expressing a baculovirus-derived trans-activator produced a 2.4-fold increase in mAb expression. The recombinant mAb was extracted from the cocoons with a buffer containing 3 m urea and purified by protein G affinity column chromatography. The antigen-binding affinity of the purified recombinant mAb was identical to that of the native mAb produced by a hybridoma. Analysis of the structure of the N-glycans attached to the recombinant mAb revealed that the mAb contained high mannose-, hybrid- and complex-type N-glycans. By contrast, insect-specific paucimannose-type glycans were not detected. Fucose residues alpha-1,3- and alpha-1,6-linked to the core N-acetylglucosamine residue, both of which are found in insect N-glycans, were not observed in the N-glycans of the mAb. PMID:19740109

  9. Gloverins of the silkworm Bombyx mori: Structural and binding properties and activities

    PubMed Central

    Yi, Hui-Yu; Deng, Xiao-Juan; Yang, Wan-Ying; Zhou, Cong-Zhao; Cao, Yang; Yu, Xiao-Qiang

    2013-01-01

    Gloverins are basic, glycine-rich and heat-stable antibacterial proteins (~14-kDa) in lepidopteran insects with activity against Escherichia coli, Gram-positive bacteria, fungi and a virus. Hyalophora gloveri gloverin adopts a random coil structure in aqueous solution but has ?-helical structure in membrane-like environment, and it may interact with the lipid A moiety of lipopolysaccharide (LPS). Manduca sexta gloverin binds to the O-specific antigen and outer core carbohydrate of LPS. In the silkworm Bombyx mori, there are four gloverins with slightly acidic to neutral isoelectric points. In this study, we investigate structural and binding properties and activities of B. mori gloverins (BmGlvs), as well as correlations between structure, binding property and activity. Recombinant BmGlv1–4 were expressed in bacteria and purified. Circular dichroism (CD) spectra showed that all four BmGlvs mainly adopted random coli structure (>50%) in aqueous solution in regardless of pH, but contained ?-helical structure in the presence of 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), smooth and rough mutants (Ra, Rc and Re) of LPS and lipid A. Plate ELISA assay showed that BmGlvs at pH 5.0 bound to rough mutants of LPS and lipid A but not to smooth LPS. Antibacterial activity assay showed that positively charged BmGlvs (at pH 5.0) were active against E. coli mutant strains containing rough LPS but inactive against E. coli with smooth LPS. Our results suggest that binding to rough LPS is the prerequisite for the activity of BmGlvs against E. coli. PMID:23567591

  10. 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

  11. Use of sleeve nets to improve survival of the Boisduval silkworm, Anaphe panda, in the Kakamega forest of western Kenya.

    PubMed

    Mbahin, N; Raina, S K; Kioko, E N; Mueke, J M

    2010-01-01

    Prospects for development of a wild silk industry in Africa would be improved if silkworm survival during mass production could be improved. A study on the survival of the Boisduval silkworm, Anaphe panda (Boisduval) (Lepidoptera: Thaumetopoeidae) was conducted with and without protection by net sleeves in two different forest habitats (natural and modified) in the Kakamega forest of western Kenya. Overall, cohort survival was significantly higher (P < 0.001) in the natural than in the modified forest, but larval survival was improved over three-fold by protection with net sleeves in both habitat types. In the modified forest, only 16.8% of unprotected larvae survived to the pupal stage and formed cocoons, whereas 62.3% survived in the same environment when they were protected with net sleeves. In the natural forest, 20.4% of unprotected larvae survived, whereas 67.7% survived in net sleeves. There was also a significant effect of season; cohorts of larvae that eclosed in the wet season had significantly lower survival than those eclosing in the dry season (P = 0.02). Sources of mortality appeared to be natural enemies (parasites, predators and diseases) and climatic factors. PMID:20569137

  12. A short-type peptidoglycan recognition protein from the silkworm: expression, characterization and involvement in the prophenoloxidase activation pathway.

    PubMed

    Chen, Kangkang; Liu, Chen; He, Yan; Jiang, Haobo; Lu, Zhiqiang

    2014-07-01

    Recognition of invading microbes as non-self is the first step of immune responses. In insects, peptidoglycan recognition proteins (PGRPs) detect peptidoglycans (PGs) of bacterial cell wall, leading to the activation of defense responses. Twelve PGRPs have been identified in the silkworm, Bombyx mori, through bioinformatics analysis. However, their biochemical functions are mostly uncharacterized. In this study, we found PGRP-S5 transcript levels were up-regulated in fat body and midgut after bacterial infection. Using recombinant protein isolated from Escherichia coli, we showed that PGRP-S5 binds to PGs from certain bacterial strains and induces bacteria agglutination. Enzyme activity assay confirmed PGRP-S5 is an amidase; we also showed it is an antibacterial protein effective against both Gram-positive and -negative bacteria. Additionally, we demonstrated that specific recognition of PGs by PGRP-S5 is involved in the prophenoloxidase activation pathway. Together, these data suggest the silkworm PGRP-S5 functions as a pattern recognition receptor for the prophenoloxidase pathway initiation and as an effecter to inhibit bacterial growth as well. We finally discussed possible roles of PGRP-S5 as a receptor for antimicrobial peptide gene induction and as an immune modulator in the midgut. PMID:24508981

  13. BmICE-2 is a novel pro-apoptotic caspase involved in apoptosis in the silkworm, Bombyx mori.

    PubMed

    Yi, Hua-Shan; Pan, Cai-Xia; Pan, Chun; Song, Juan; Hu, Yan-Fen; Wang, La; Pan, Min-Hui; Lu, Cheng

    2014-02-28

    In this study we identified a potential pro-apoptotic caspase gene, Bombyx mori(B. mori)ICE-2 (BmICE-2) which encoded a polypeptide of 284 amino acid residues, including a (169)QACRG(173) sequence which surrounded the catalytic site and contained a p20 and a p10 domain. BmICE-2 expressed in Escherichia coli (E. coli) exhibited high proteolytic activity for the synthetic human initiator caspase-9 substrates Ac-LEHD-pNA, but little activity towards the effector caspase-3 substrates Ac-DEVD-pNA. When BmICE-2 was transiently expressed in BmN-SWU1 silkworm B. mori cells, we found that the high proteolytic activity for Ac-LEHD-pNA triggered caspase-3-like protease activity resulting in spontaneous cleavage and apoptosis in these cells. This effect was not replicated in Spodoptera frugiperda 9 cells. In addition, spontaneous cleavage of endogenous BmICE-2 in BmN-SWU1 cells could be induced by actinomycin D. These results suggest that BmICE-2 may be a novel pro-apoptotic gene with caspase-9 activity which is involved apoptotic processes in BmN-SWU1 silkworm B. mori cells. PMID:24491540

  14. 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

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

    SciTech Connect

    Wang Shengpeng [Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai (China); Graduate School, Chinese Academy of Sciences, Shanghai (China); Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang (China); Guo Tingqing [Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai (China); Graduate School, Chinese Academy of Sciences, Shanghai (China); Guo Xiuyang [Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai (China); Graduate School, Chinese Academy of Sciences, Shanghai (China); Huang Junting [Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang (China); Lu Changde [Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai (China)]. 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.

  16. Translationally controlled tumor protein, a dual functional protein involved in the immune response of the silkworm, Bombyx mori.

    PubMed

    Wang, Fei; Hu, Cuimei; 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

  17. Isolation, purification, and identification of an important pigment, sepiapterin, from integument of the lemon mutant of the silkworm, Bombyx mori.

    PubMed

    Gao, Junshan; Wang, Jing; Wang, Wenjing; Liu, Chaoliang; Meng, Yan

    2013-01-01

    Sepiapterin is the precursor of tetrahydrobiopterin, an important coenzyme of aromatic amino acid hydroxylases, the lack of which leads to a variety of physiological metabolic diseases or neurological syndromes in humans. Sepiapterin is a main pigment component in the integument of the lemon mutant of the silkworm, Bombyx mori (L.) (Lepidoptera: Bombycidae), and is present there in extremely high content, so lemon is a valuable genetic resource to extract sepiapterin. In this study, an effective experimental system was set up for isolation and purification of sepiapterin from lemon silkworms by optimizing homogenization solvent, elution buffer, and separation chromatographic column. The results showed that ethanol was the most suitable solvent to homogenize the integument, with a concentration of 50% and solid:liquid ratio of 1:20 (g/mL). Sepiapterin was purified successively by column chromatography of cellulose Ecteola, sephadex G-25-150, and cellulose phosphate, and was identified by ultraviolet-visible absorption spectrometry. A stable and accurate high performance liquid chromatography method was constructed to identify sepiapterin and conduct qualitative and quantitative analyses. Sepiapterin of high purity was achieved, and the harvest reached about 40 ug/g of integument in the experiments. This work helps to obtaining natural sepiapterin in large amounts in order to use the lemon B. mori mutant to produce BH4 in vitro. PMID:24773269

  18. Germline transformation of the silkworm Bombyx mori L. using a piggyBac transposon-derived vector.

    PubMed

    Tamura, T; Thibert, C; Royer, C; Kanda, T; Abraham, E; Kamba, M; Komoto, N; Thomas, J L; Mauchamp, B; Chavancy, G; Shirk, P; Fraser, M; Prudhomme, J C; Couble, P; Toshiki, T; Chantal, T; Corinne, R; Toshio, K; Eappen, A; Mari, K; Natuo, K; Jean-Luc, T; Bernard, M; Gérard, C; Paul, S; Malcolm, F; Jean-Claude, P; Pierre, C

    2000-01-01

    We have developed a system for stable germline transformation in the silkworm Bombyx mori L. using piggyBac, a transposon discovered in the lepidopteran Trichoplusia ni. The transformation constructs consist of the piggyBac inverted terminal repeats flanking a fusion of the B. mori cytoplasmic actin gene BmA3 promoter and the green fluorescent protein (GFP). A nonautonomous helper plasmid encodes the piggyBac transposase. The reporter gene construct was coinjected into preblastoderm eggs of two strains of B. mori. Approximately 2% of the individuals in the G1 broods expressed GFP. DNA analyses of GFP-positive G1 silkworms revealed that multiple independent insertions occurred frequently. The transgene was stably transferred to the next generation through normal Mendelian inheritance. The presence of the inverted terminal repeats of piggyBac and the characteristic TTAA sequence at the borders of all the analyzed inserts confirmed that transformation resulted from precise transposition events. This efficient method of stable gene transfer in a lepidopteran insect opens the way for promising basic research and biotechnological applications. PMID:10625397

  19. 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. PMID:24632065

  20. Translational enhancement of recombinant protein synthesis in transgenic silkworms by a 5'-untranslated region of polyhedrin gene of Bombyx mori Nucleopolyhedrovirus.

    PubMed

    Iizuka, Masashi; Tomita, Masahiro; Shimizu, Katsuhiko; Kikuchi, Yutaka; Yoshizato, Katsutoshi

    2008-06-01

    Previously, we established a method to produce recombinant proteins (r-proteins) in cocoons of germline transgenic silkworms, and showed that a step(s) in post-transcription processes was rate-limiting in obtaining a high yield of r-proteins. In this study, we examined whether the 5'-untranslated region (5'-UTR) of the polyhedrin gene (pol) of nucleopolyhedrovirus (NPV) has a translational enhancer activity in the r-protein expression by middle silk gland (MSG) cells of silkworm Bombyx mori (Bm). Sericin 1 gene (ser1) promoter-driven transformation vectors were constructed in which pol5'-UTRs of NPVs isolated from four different species, Bm, Spodoptera frugiperda, Ectropis oblique, and Malacosoma neustria, were each placed upstream of a reporter gene. Transient expression assays in MSGs showed that these pol5'-UTRs all enhanced the protein expression of reporter genes, and the pol5'-UTR of Bm NPV (pol5'-UTR/Bm) was the most effective among them. Thus, transgenic silkworms were generated, which bore the ser1 promoter-driven His-tagged secretory EGFP (sEGFP-His) gene under the control of pol5'-UTR/Bm. The synthesis of sEGFP-His proteins in MSGs of the transgenic worms was approximately 1.5-fold higher than that in those bearing null vectors. However, its mRNA expression levels were 67% of the control worms, indicating that the pol5'-UTR/Bm specifically enhanced the translational level. In conclusion, pol5'-UTR/Bm increased the yield of r-protein production in transgenic silkworms by enhancing the translational activity and this 5'-UTR could be useful for the mass production of r-proteins in germline transgenic silkworms. PMID:18640598

  1. Major Oak Diseases and Their Control. 

    E-print Network

    Johnson, Jerral D.; Appel, David N.

    1984-01-01

    )OC ?A245 Q7 173 . \\00 ILl78 , LIBRARY -?'?AUG 3 1?1S84 r ex.J$ MM University MAJOR OAK DISEASES AND THEIR CONTROL 8-1478 J ~ Texas Agricultural Extension Service. The Texas A&M University System ? Zerle L. Carpenter, Director. College... Station, Texas (Blaok Page bl-?OriglulBidletial? / . , ,;..,' ,- ; ~ ~ " DISEASES AND THEIR CONTROL Jerral D. Johnson and David N. Appel* Oaks represent the major shade trees of Texas and are also...

  2. Oak Ridge Reservation environmental report for 1989

    SciTech Connect

    Jacobs, V.A.; Wilson, A.R. (eds.)

    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.

  3. Non-oak native plants are main hosts for sudden oak death pathogen in California

    Microsoft Academic Search

    Matteo Garbelotto; Jennifer M. Davidson; Kelly Ivors; Patricia E. Maloney; Daniel Hüberli; Steven T. Koike; David M. Rizzo

    2002-01-01

    The finding of Phytophthora ramorum — the pathogen that causes sudden oak death in four California native trees — on rhododendron in Europe led us to hypothesize that its host range in California's natural forests was much greater than previously suspected. In addition to the affected oak species, we have now identified an additional 13 species from 10 plant families

  4. An aerial radiological survey of the Oak Ridge Reservation and surrounding area, Oak Ridge, Tennessee

    Microsoft Academic Search

    1989-01-01

    An aerial radiological survey of the Oak Ridge Reservation (ORR) and surrounding area in Oak Ridge, Tennessee, was conducted from September 12--29, 1989. The purpose of the survey was to measure and document the site's terrestrial radiological environment for use in effective environmental management and emergency response planning. The aerial survey was flown at an altitude of 91 meters (300

  5. A study of shin oak (Quercus havardi) poisoning in cattle 

    E-print Network

    Dollahite, James Walton

    1961-01-01

    poisoned bp feeding cake TABLE i)F COPTERTS I IRTRi)DGCTION II?REVIEW OF TBE LITERATORE Ill. MATE', R)AIS AND METHODS A. Feeding Shin Oak to Calves B. Feeding Shin Oak to Sheep C. Feeding Shin Oak to Goa'ts D. Feeding Shin Oak to Guinea Pigs E... of the state. The buds and young jeaves of rriany of the oaks are considered to be poisonous vtren eaten exclusively by livestock. The low-growing fo'rms of shir. oaks usually cause the most trouble with the species Quercus havardi considered to be 'he most...

  6. Morphogenesis of the antenna of the male silkmoth. Antheraea polyphemus, III. Development of olfactory sensilla and the properties of hair-forming cells.

    PubMed

    Keil, T A; Steiner, C

    1991-01-01

    During adult development of the male silkmoth Antheraea polyphemus, the anlagen of olfactory sensilla arise within the first 2 days post-apolysis in the antennal epidermis (stage 1-3). Approximately on the second day, the primary dendrites as well as the axons grow out from the sensory neurons (stage 4). The trichogen cells start to grow apical processes approximately on the third day, and these hair-forming 'sprouts' reach their definite length around the ninth day (stages 5-6). Then the secretion of cuticle begins, the cuticulin layer having formed on day 10 (stage 7a). The primary dendrites are shed, the inner dendritic segments as well as the thecogen cells retract from the prospective hair bases, and the inner tormogen cells degenerate around days 10/11 (stage 7b). The hair shafts of the basiconic sensilla are completed around days 12/13 (stage 7c), and those of the trichoid sensilla around days 14/15 (stage 7d). The trichogen sprouts retract from the hairs after having finished cuticle formation, and the outer dendritic segments grow out into the hairs: in the basiconic sensilla directly through, and in the trichoid sensilla alongside, the sprouts. The trichogen sprouts contain numerous parallel-running microtubules. Besides their cytoskeletal function, these are most probably involved in the transport of membrane vesicles. During the phase of cuticle deposition, large numbers of vesicles are transported anterogradely from the cell bodies into the sprouts, where they fuse with the apical cell membrane and release their electron-dense contents (most probably cuticle precursors) to the outside. As the cuticle grows in thickness, the surface area of the sprouts is reduced by endocytosis of coated vesicles. When finally the sprouts retract from the completed hairs, the number of endocytotic vesicles is further increased and numerous membrane cisterns seem to be transported retrogradely along the microtubules to the cell bodies. Here the membrane material will most probably be used again in the formation of the sensillum lymph cavities. Thus, the trichogen cells are characterized by an intensive membrane recycling. The sensillum lymph cavities develop between days 16-20 (stage 8), mainly via apical invaginations of the trichogen cells. The imago emerges on day 21. PMID:18621189

  7. Structural insights into the ligand binding and releasing mechanism of Antheraea polyphemus pheromone-binding protein 1: role of the C-terminal tail.

    PubMed

    Katre, Uma V; Mazumder, Suman; Mohanty, Smita

    2013-02-12

    Pheromone-binding proteins (PBPs) in lepidopteran moths selectively transport the hydrophobic pheromone molecules across the sensillar lymph to trigger the neuronal response. Moth PBPs are known to bind ligand at physiological pH and release it at acidic pH while undergoing a conformational change. Two molecular switches are considered to play a role in this mechanism: (i) protonation of His(70) and His(95) situated at one end of binding pocket and (ii) switch of the unstructured C-terminus at the other end of the binding pocket to a helix that enters the pocket. We have reported previously the role of the histidine-driven switch in ligand release for Antheraea polyphemus PBP1 (ApolPBP1). Here we show that the C-terminus plays a role in the ligand release and binding mechanism of ApolPBP1. The C-terminus truncated mutants of ApolPBP1 (ApolPBP1?P129-V142 and ApolPBP1H70A/H95A?P129-V142) exist only in the bound conformation at all pH levels, and they fail to undergo pH- or ligand-dependent conformational switching. Although these proteins could bind ligands even at acidic pH unlike wild-type ApolPBP1, they had ~4-fold reduced affinity for the ligand at both acidic and physiological pH compared to that of wild-type ApolPBP1 and ApolPBP1H70A/H95A. Thus, apart from helping in ligand release at acidic pH, the C-terminus in ApolPBP1 also plays an important role in ligand binding and/or locking the ligand in the binding pocket. Our results are in stark contrast to those reported for BmorPBP and AtraPBP, where C-terminus truncated proteins had similar or increased pheromone binding affinity at any pH. PMID:23327454

  8. Structural Insights into the Ligand Binding and Releasing Mechanism of Antheraea polyphemus PBP1: Role of the C-terminal Tail

    PubMed Central

    Katre, Uma V.; Mazumder, Suman; Mohanty, Smita

    2013-01-01

    Pheromone-binding proteins (PBPs) in lepidopteran moths selectively transport the hydrophobic pheromone molecules across the sensillar lymph to trigger the neuronal response. Moth PBPs are known to bind ligand at physiological pH and release it at acidic pH while undergoing a conformational change. Two molecular switches are considered to play a role in this mechanism: (i) Protonation of His70 and His95 situated at one end of binding pocket, and (ii) Switch of the unstructured C-terminus at the other end of the binding pocket to a helix that enters the pocket. We have reported previously the role of the histidine-driven switch in ligand release for Antheraea polyphemus PBP1 (ApolPBP1). Here we show that the C-terminus plays a role in ligand release and binding mechanism of ApolPBP1. The C-terminus truncated mutants of ApolPBP1 (ApolPBP1?P129-V142 and ApolPBP1H70A/H95A?P129-V142) exist only in the bound conformation at all pH levels, and they fail to undergo pH- or ligand- dependent conformational switch. Although these proteins could bind ligands even at acidic pH unlike the wild-type ApolPBP1, they had ~4 fold reduced affinity towards the ligand at both acidic and physiological pH than that of ApolPBP1wt and ApolPBP1H70A/H95A. Thus, apart from helping in the ligand-release at acidic pH, the C-terminus in ApolPBP1 also plays an important role in ligand binding and/or locking the ligand in the binding pocket. Our results are in stark contrast to those reported for BmorPBP and AtraPBP, where C-terminus truncated proteins had similar or increased pheromone-binding affinity at any pH. PMID:23327454

  9. 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.

  10. 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 children and their families, "Working Among the Oaks" has focused on connecting with the agricultural and environmental communities. For example, the Ranching Sustainability Self-Assessment Program is an ambitious, long-range project with tremendous potential to aid private landowners throughout California in implementing sustainable ranching practices. We've made great progress through the efforts of an impressive committee of local private landowners, ranch managers and resource professionals. They believe that this can be a powerful non-regulatory tool to guide private landowners through everyday decision-making processes. Most importantly, this is a tool that could be adapted for use throughout California oak woodland. The Self Assessment Program, along with the supporting Workshops, have stimulated discussion and interest in sustainable ranching among people with diverse experiences and backgrounds. "Learning and Working Among the Oaks" together reach the full spectrum of oak conservation stakeholders, from kids to grandparents, town residents to ranching families, environmental groups to farm and vineyard managers, and more. The diversity of these stakeholders helps us identify collaborative education and research opportunities to support education and management of the 3 million ha of California oak woodlands.

  11. A dense genetic map of the silkworm, Bombyx mori, covering all chromosomes based on 1018 molecular markers.

    PubMed

    Yasukochi, Y

    1998-12-01

    A dense linkage map was constructed for the silkworm, Bombyx mori, containing 1018 genetic markers on all 27 autosomes and the Z chromosome. Most of the markers, covering approximately 2000 cM, were randomly amplified polymorphic DNAs amplified with primer-pairs in combinations of 140 commercially available decanucleotides. In addition, eight known genes and five visible mutations were mapped. Bombyx homologues of engrailed and invected genes were found to be closely linked, as in Drosophila melanogaster. The average interval between markers was approximately 2 cM, equal to approximately 500 kb. The correspondence of seven linkage groups to counterparts of the conventional linkage map was determined. This map is the first linkage map in insects having a large number of chromosomes (n = 28) that covers all chromosomes without any gaps. PMID:9832528

  12. Free radical scavenging and tyrosinase inhibition activity of oils and sericin extracted from Thai native silkworms (Bombyx mori).

    PubMed

    Manosroi, Aranya; Boonpisuttinant, Korawinwich; Winitchai, Supanida; Manosroi, Worapaka; Manosroi, Jiradej

    2010-08-01

    Oils and sericin were extracted from pupae and silk cocoons, respectively, of the five Thai native silkworms (Bombyx mori, Linnaeus (Bombycidae)), namely, Keawsakol, Nangnoi, Somrong, Nangleung, and Noneruesee, which are variations of the same species. The oils were extracted by a hot process using Soxhlet apparatus and a cold process using petroleum ether, while sericin was extracted by basic hydrolysis and autoclaving. Sericin from the five Thai native silkworms showed free radical scavenging activity lower than the standard antioxidants (vitamin C, vitamin E, and BHT) by about 20-100-fold, but all oils gave higher activity than that of the standard linoleic acid by 11-22-fold. Oil extracted from Noneruesee by the cold process gave the highest DPPH scavenging activity, compared with other oil samples. All sericin samples showed tyrosinase inhibition activity with IC(50) values in the range of 1.2-18.76 mg/mL, but only oils from Noneruesee extracted by the hot process, and Nangleung, Somrong, and Noneruesee extracted by the cold process, showed this activity. Oil extracted by the hot process and sericin by basic hydrolysis from Noneruesee gave the highest tyrosinase inhibition activity, but lower than that of the standards vitamin C and kojic acid by 20-49 and 3-8 times, respectively. This study has suggested that sericin and oil from Noneruesee extracted by basic hydrolysis and the cold process, which gave the highest tyrosinase inhibition and free radical scavenging activity, respectively, can be applied in antiaging and whitening cosmetic products. PMID:20673171

  13. [Construction of AFLP molecular markers linkage map and localization of green cocoon gene in silkworm (Bombyx mori)].

    PubMed

    Zhao, Ai-Chun; Lu, Cheng; Li, Bin; Pu, Xiao-Ying; Zhou, Ze-Yang; Xiang, Zhong-Huai

    2004-08-01

    Based on an improved method of AFLP, AFLP markers were employed for construction of a linkage map and localization of Gc gene used a set of 44 backcross lines( BC1) of silkworm ( Bombyx mori) as a mapping population. In this work, all together 3 956 bands were obtained by 28 pairs of primers and 141.3 bands each primer pair on average. Among them 2 836 bands were in good agreement with the segregation pattern. A total of 1 018 (25.7%) polymorphic AFLP markers were detected. The 693 (68.1%) of polymorphic markers with 1:1 segregation ratio ( P < or = 0.05) were obtained. Furthermore,The analytical model was based on the backcross type and the parameters were set as following: LOD = 3.0, maximum recombination value of 0. 20 and use the command ' group', 'compare', 'try', 'map' and 'ripple' to construct the linkage maps. 407 of the 693 loci were chi2 tested in agreement with 1:1 segregation were divided into 33 linkages by Mapmaker/Exp(Version 3.0), with a total map distance of 3 676.7 cM and a mean distance of 9.1 cM between markers. The morphological gene Gc was located between L-P4T6-107 and L-PT6T4-84 on linkage group 22. In addition, 286 markers were not included in the linkage groups. The efficiency of loci mapping was 58.7%. Among the 33 linkage groups, the morphological marker Gc classically localized on linkage group 15 was relocated on linkage group 22 on the map, suggesting that this molecular linkage group corresponds to linkage group 15 on the linkage map based on morphological characters. All these have laid an important base for the marker assisted breeding of the silkworm. PMID:15481532

  14. 2. ARROYO SECO PARKWAY AND FAIR OAKS BRIDGE OVERCROSSING. ABANDONED ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    2. ARROYO SECO PARKWAY AND FAIR OAKS BRIDGE OVERCROSSING. ABANDONED RAILROAD TRESTLE IN REAR. LOOKING 244°WSW. - Arroyo Seco Parkway, Fair Oaks Avenue Bridge, Milepost 31.17, Los Angeles, Los Angeles County, CA

  15. Poison Ivy, Oak and Sumac Rashes Can Be Serious

    MedlinePLUS

    ... nlm.nih.gov/medlineplus/news/fullstory_152188.html Poison Ivy, Oak and Sumac Rashes Can Be Serious ... 23, 2015 (HealthDay News) -- Itchy, blistering rashes from poison ivy, oak and sumac are common and are ...

  16. 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

  17. Oak dune forests in Denmark and their ecology

    Microsoft Academic Search

    Jonas E Lawesson; Peter Wind

    2002-01-01

    Floristic and structural data of oak dune forests in western Jutland (Kaergård and Blåbjerg) were collected in 1998 and compared with previous studies of Danish oligotrophic oak forests.A hierarchical divisive clustering resulted in four main clusters, containing two coastal oak vegetation types, Quercus robur–Carex arenaria and Q. robur–Pleurozium schreberi communities, both restricted to the oak dune forest areas in western

  18. A novel angiotensin-? 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 ?-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 102.15?M, 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. PMID:25111373

  19. Author's personal copy Environmentally biased fragmentation of oak savanna

    E-print Network

    Vellend, Mark

    Author's personal copy Environmentally biased fragmentation of oak savanna habitat on southeastern Available online 7 September 2008 Keywords: Climate Conservation baselines Indigenous land use Oak savanna environmental conditions represented across the entire landscape, in oak savanna habitats prior to Euro- pean

  20. Blue oak seedlings may be older than they look

    E-print Network

    Standiford, Richard B.

    Blue oak seedlings may be older than they look Ralph L. Phillips u Neil K. McDougald o Richard B. Standiford William E.Frost A 4-year study indicates that na- tive blue oak seedlings are prob- ably much the year of above- average rainfall. Blue oak (Quercusdouglasii)trees are a valuable economic and aesthetic

  1. Generation of a transgenic silkworm that secretes recombinant proteins in the sericin layer of cocoon: Production of recombinant human serum albumin

    Microsoft Academic Search

    Shingo Ogawa; Masahiro Tomita; Katsuhiko Shimizu; Katsutoshi Yoshizato

    2007-01-01

    In this study we produced germline transgenic silkworms that spin cocoons containing recombinant human serum albumin (rHSA) in the sericin layer. A piggyBac-based transformation vector was constructed that carried HSA cDNA driven by sericin-1 gene promoter, viral enhancer hr3, and gene encoding viral trans-activator IE1. Isolated silk glands were bombarded with the vector and transplanted into host larvae. Three days

  2. Production of a non-triple helical collagen alpha chain in transgenic silkworms and its evaluation as a gelatin substitute for cell culture.

    PubMed

    Adachi, Takahiro; Wang, Xiaobiao; Murata, Tomoko; Obara, Masanobu; Akutsu, Hidenori; Machida, Masakazu; Umezawa, Akirihiro; Tomita, Masahiro

    2010-08-15

    We generated transgenic silkworms that synthesized human type I collagen alpha1 chain [alpha1(I) chain] in the middle silk glands and secreted it into cocoons. The initial content of the recombinant alpha1(I) chain in the cocoons of the transgenic silkworms was 0.8%. The IE1 gene, a trans-activator from the baculovirus, was introduced into the transgenic silkworm to increase the content of the chain. We also generated silkworms homozygous for the transgenes. These manipulations increased the alpha1(I) chain content to 8.0% (4.24 mg per cocoon). The alpha1(I) chain was extracted and purified from the cocoons using a very simple method. The alpha1(I) chain contained no hydroxyprolines due to the absence of prolyl-hydroxylase activity in the silk glands. Circular dichroism analysis showed that the secondary structure of the alpha1(I) chain is similar to that of denatured type I collagen, demonstrating the absence of the triple helical structure. Human skin fibroblasts were seeded on the alpha1(I) chain-coated dishes. The cells attached and spread, although at decreased chain concentrations the spreading rate was lower than that of the collagen and gelatin. Cynomolgus monkey embryonic stem cells cultured on the alpha1(I) chain-coated dishes maintained an undifferentiated state after 30 passages, and their pluripotency was confirmed by teratoma formation in severe combined immunodeficient mice. These results show that the recombinant human alpha1(I) chain is a promising candidate biomaterial as a high-quality and safe gelatin substitute for cell culture. PMID:20589667

  3. Studies on middle silkgland proteins of cocoon colour sex-limited silkworm ( Bombyx mori L.) using two-dimensional polyacrylamide gel electrophoresis

    Microsoft Academic Search

    Yuan-Xiang Jin; Yu-Yin Chen; Meng-Kui Xu; Yong-Huang Jiang

    2004-01-01

    Qualitative and quantitative differences in proteins expressed in the middle silkglands of male and female silkworm larvae\\u000a that differ in silk colour were investigated by high resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE),\\u000a followed by computer assisted image analysis. About 1000 protein spots were resolved in both the sexes and most proteins were\\u000a shown to be distributed in the area

  4. Genetic diversity and phylogenetic relationships among microsporidia infecting the silkworm, Bombyx mori, using random amplification of polymorphic DNA: Morphological and ultrastructural characterization

    Microsoft Academic Search

    S. Nageswara Rao; B. Surendra Nath; G. Bhuvaneswari; S. Raje Urs

    2007-01-01

    Random amplification of polymorphic DNA polymerase chain reaction (RAPD-PCR) and pathological, morphological and ultrastructural characterization were used to differentiate seven new microsporidian isolates infecting the mulberry silkworm, Bombyx mori. The pathogenicity observed was dose-dependent and differed from each of the microsporidian isolates; the NIK-4m was found to be more virulent than other isolates. However, all the isolates, except NIK-4m, showed

  5. Molecular properties and tissue distribution of 30K proteins as ommin-binding proteins from diapause eggs of the silkworm, Bombyx mori

    Microsoft Academic Search

    Hiroshi Sawada; Yumi Yamahama; Keisuke Mase; Haruka Hirakawa; Teruhiko Iino

    2007-01-01

    We previously reported the purification of an ommin-binding protein (OMBP) from an acid-methanol extract of diapause eggs of the silkworm and that OMBP reacted with the anti-30K proteins antiserum. In order to clarify the relationship between OMBP and the 30K proteins, we attempted to determine the sequence of the N-terminal amino acid of OMBP, which was separated by two-dimensional polyacrylamide

  6. The isolation and evaluation of endophytic bacteria from live oaks as potential biological control agents for oak wilt in Texas 

    E-print Network

    Brooks, David Stewart

    1989-01-01

    were isolated from live oaks (Quercus fusiformis) at three oak wilt centers in central Texas over a one year period. The bacterial isolates were evaluated as potential biological control agents of oak wilt. A total of 889 bacterial isolates were... that bacterial isolates tested were better able to distribute both above and below the point of inoculation than in the fall study. Challenge experiments were conducted to evaluate the ability of selected bacterial isolates to inhibit oak wilt development...

  7. 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%), ?-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 ?-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

  8. Morphological and molecular studies of Vairimorpha necatrix BM, a new strain of the microsporidium V. necatrix (Microsporidia, Burenellidae) recorded in the silkworm, Bombyx mori.

    PubMed

    Luo, Bo; Liu, Handeng; Pan, Guoqing; Li, Tian; Li, Zeng; Dang, Xiaoqun; Liu, Tie; Zhou, Zeyang

    2014-08-01

    Vairimorpha sp. BM (2012) is a recent isolate of the microsporidia from the silkworm in Shandong, China. The ultrastructure, tissue pathology and molecular characterization of this isolate is described in this study. This pathogenic fungus causes pebrine disease in silkworms which manifests as a systemic infection. Meanwhile, the silkworm eggs produced by the infected moths were examined using a microscope and PCR amplification. Neither spores nor the expected PCR band were observed, suggesting that no vertical transmission occurred in Bombyx mori. In addition, the ultrastructure of the isolate was studied by light microscopy and transmission electron microscopy. Two types of spores were observed: diplokaroytic spores with 13-17 coils of polar tubes and monokaryotic spores with less coils of polar tubes which could form octospores; however, no sporophorous vesicles were observed. Finally, phylogenetic analysis of the small subunit rRNA genes of Vairimorpha species showed that this isolate has a closer relationship to Vairimorpha necatrix than the other species studied. This result also is supported by phylogenetic analysis based on their actin genes, heat shock protein 70 (HSP70) and RNA polymerase II (RPB1). Based on the information gained during this study, we propose that this microsporidian species infecting B. mori should be given the name V. necatrix BM. PMID:24818618

  9. 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. PMID:22135907

  10. 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 ?g 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 8.11-folds. Transcription levels of nine genes were further verified by RT-qPCR, and the results were consistent with those with DGE. Transcription and expression levels of fibroin light chain (Fib-L) gene were increased after TiO2 NPs feeding, indicating that TiO2 NPs improves fibroin synthesis. Compared with that of control, the mean protease activity was increased by 56.67 % in the B. mori fed with TiO2 NPs, and the transport of four key amino acids used for fibroin synthesis in hemolymph was also increased. These findings indicated that TiO2 NPs feeding can improve the absorption and utilization of amino acids from the feed and could be a new way to increase the fibroin synthesis in B. mori. PMID:25876086

  11. Dendrochronological parameters of northern red oak ( Quercus rubra L. (Fagaceae)) infested with red oak borer ( Enaphalodes rufulus (Haldeman) (Coleoptera: Cerambycidae))

    Microsoft Academic Search

    L. J. Haavik; F. M. Stephen; M. K. Fierke; V. B. Salisbury; S. W. Leavitt; S. A. Billings

    2008-01-01

    Oak-dominated forests in northwestern Arkansas have recently experienced an oak mortality event associated with an unprecedented outbreak of a native insect, the red oak borer, Enaphalodes rufulus (Haldeman). To determine whether prior drought was associated with increased E. rufulus infestation level of Quercus rubra L. trees, we employed a suite of dendrochronological measurements from Q. rubra in affected forest stands.

  12. Oak Ridge National Laboratory 5-1 5. Oak Ridge National Laboratory

    E-print Network

    Pennycook, Steve

    of the secret Manhattan Project to pioneer a method for producing and separating plutonium. During the 1950s, but very different from, the work carried out in the days of the Manhattan Project. #12;Oak Ridge

  13. White Oak Creek embayment sediment retention structure: The Oak Ridge model in action

    Microsoft Academic Search

    S. D. Van Hoesen; B. L. Kimmel; D. G. Page; G. R. Hudson; R. B. Wilkerson; M. Zocolla; J. L. Kauschinger

    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

  14. Design assessment for the Bethel Valley FFA Upgrades at Oak Ridge National Laboratory, Oak Ridge, Tennessee

    SciTech Connect

    NONE

    1995-09-01

    This report describes the proposed upgrades to Building 3025 and the Evaporator Area at Oak Ridge National Laboratory. Design assessments, specifications and drawings are provided. Building 3025 is a general purpose research facility utilized by the Materials and Ceramics Division to conduct research on irradiated materials. The Evaporator Area, building 2531, serves as the collection point for all low-level liquid wastes generated at the Oak Ridge National Laboratory.

  15. Original article Oak tree improvement in Indiana

    E-print Network

    Paris-Sud XI, Université de

    Original article Oak tree improvement in Indiana MV Coggeshall Indiana Department of Natural of Indiana is currently in progress for 4 species: Quercus rubra L, Q alba, Q falcata var pagodifolia Ell testing Résumé — Programmes d'amélioration des chênes dans l'État d'Indiana. Des programmes d

  16. ORNL (Oak Ridge National Laboratory) 89

    Microsoft Academic Search

    T. D. Anderson; B. R. Appleton; J. W. Jefferson; J. R. Merriman; F. R. Mynatt; C. R. Richmond; M. W. Rosenthal

    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

  17. 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.

  18. Seismic hazard evaluation for Department of Energy Oak Ridge Reservations, Oak Ridge, Tennessee

    SciTech Connect

    McGuire, R.K.; Toro, G.F. [Risk Engineering, Inc., Golden, CO (United States); Hunt, R.J. [Martin Marietta Energy Systems, Inc., Oak Ridge, TN (United States). Center for Natural Phenomena Engineering

    1992-09-30

    This study presents the results of an investigation of seismic hazard at the Department of Energy Oak Ridge Reservations (K-25 Site, Oak Ridge National Laboratories, and Oak Ridge Y-12 Plant), located in Oak Ridge, Tennessee. Oak Ridge is located in eastern Tennessee, in an area of moderate to high historical seismicity. Results from two separate seismic hazard analyses are presented. The EPRI/SOG analysis uses the input data and methodology developed by the Electric Power Research Institute, under the sponsorship of several electric utilities, for the evaluation of seismic hazard in the central and eastern United States. The LLNL analysis uses the input data and methodology developed by the Lawrence Livermore National Laboratory for the Nuclear Regulatory Commission. Both the EPRI/SOG and LLNL studies characterize earth-science uncertainty on the causes and characteristics of earthquakes in the central and eastern United States. This is accomplished by considering multiple hypotheses on the locations and parameters of seismic source zones and by considering multiple attenuation functions for the prediction of ground shaking given earthquake size and location. These hypotheses were generated by multiple expert teams and experts. Furthermore, each team and expert was asked to generate multiple hypotheses in order to characterize his own internal uncertainty. The seismic-hazard calculations are performed for all hypotheses. Combining the results from each hypothesis with the weight associated to that hypothesis, one obtains an overall representation of the seismic hazard at the Oak Ridge site and its uncertainty.

  19. LIM-homeodomain transcription factor Awh is a key component activating all three fibroin genes, fibH, fibL and fhx, in the silk gland of the silkworm, Bombyx mori.

    PubMed

    Kimoto, Mai; Tsubota, Takuya; Uchino, Keiro; Sezutsu, Hideki; Takiya, Shigeharu

    2015-01-01

    In the silkworm Bombyx mori, three fibroin genes, fibroin-heavy-chain (fibH), fibroin-light-chain (fibL) and fibrohexamerin (fhx), are coexpressed only in the posterior silk gland (PSG) cells, while the sericin genes encoding silk glue proteins are expressed in the middle silk gland (MSG) cells. Silk gland factor-2 (SGF-2) is a PSG-specific activator complex of fibH, composed of a LIM-homeodomain protein, Awh, and its cofactors, Ldb and Lcaf. We investigated whether SGF-2 can activate other fibroin genes using transgenic silkworms. The genes for Ldb and Lcaf were expressed ubiquitously in various tissues, while the gene for Awh was expressed strictly specific in PSG of the wild type silkworms. Misexpression of Awh in transgenic silkworms induced ectopic expression of fibL and fhx as well as fibH in MSG. Coincidently with the induction of fibL and fhx by Awh, binding of SGF-2 to the promoter of fibL and fhx was detected in vitro, and SGF-2 binds directly to the fhx core promoter. Ectopic expression of the fibroin genes was observed at high levels in the middle part of MSG. Moreover, fibL and fhx were induced in the anterior silk gland (ASG) of the transgenic silkworms, but fibH was not. These results indicate that Awh is a key activator of all three fibroin genes, and the activity is probably regulated in conjunction with additional factors. PMID:25449130

  20. 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

    NONE

    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.

  1. Osteochondral Tissue Engineering In Vivo: A Comparative Study Using Layered Silk Fibroin Scaffolds from Mulberry and Nonmulberry Silkworms

    PubMed Central

    Saha, Sushmita; Kundu, Banani; Kirkham, Jennifer; Wood, David; Kundu, Subhas C.; Yang, Xuebin B.

    2013-01-01

    The ability to treat osteochondral defects is a major clinical need. Existing polymer systems cannot address the simultaneous requirements of regenerating bone and cartilage tissues together. The challenge still lies on how to improve the integration of newly formed tissue with the surrounding tissues and the cartilage-bone interface. This study investigated the potential use of different silk fibroin scaffolds: mulberry (Bombyx mori) and non-mulberry (Antheraea mylitta) for osteochondral regeneration in vitro and in vivo. After 4 to 8 weeks of in vitro culture in chondro- or osteo-inductive media, non-mulberry constructs pre-seeded with human bone marrow stromal cells exhibited prominent areas of the neo tissue containing chondrocyte-like cells, whereas mulberry constructs pre-seeded with human bone marrow stromal cells formed bone-like nodules. In vivo investigation demonstrated neo-osteochondral tissue formed on cell-free multi-layer silk scaffolds absorbed with transforming growth factor beta 3 or recombinant human bone morphogenetic protein-2. Good bio-integration was observed between native and neo-tissue within the osteochondrol defect in patellar grooves of Wistar rats. The in vivo neo-matrix formed comprised of a mixture of collagen and glycosaminoglycans except in mulberry silk without growth factors, where a predominantly collagenous matrix was observed. Immunohistochemical assay showed stronger staining of type I and type II collagen in the constructs of mulberry and non-mulberry scaffolds with growth factors. The study opens up a new avenue of using inter-species silk fibroin blended or multi-layered scaffolds of a combination of mulberry and non-mulberry origin for the regeneration of osteochondral defects. PMID:24260335

  2. Oral Administration of a Fusion Protein between the Cholera Toxin B Subunit and the 42-Amino Acid Isoform of Amyloid-? Peptide Produced in Silkworm Pupae Protects against Alzheimer's Disease in Mice

    PubMed Central

    Li, Si; Wei, Zhen; Chen, Jian; Chen, Yanhong; Lv, Zhengbing; Yu, Wei; Meng, Qiaohong; Jin, Yongfeng

    2014-01-01

    A key molecule in the pathogenesis of Alzheimer's disease (AD) is a 42-amino acid isoform of the amyloid-? peptide (A?42), which is the most toxic element of senile plaques. In this study, to develop an edible, safe, low-cost vaccine for AD, a cholera toxin B subunit (CTB)-A?42 fusion protein was successfully expressed in silkworm pupae. We tested the silkworm pupae-derived oral vaccination containing CTB-A?42 in a transgenic mouse model of AD. Anti-A?42 antibodies were induced in these mice, leading to a decreased A? deposition in the brain. We also found that the oral administration of the silk worm pupae vaccine improved the memory and cognition of mice, as assessed using a water maze test. These results suggest that the new edible CTB-A?42 silkworm pupae-derived vaccine has potential clinical application in the prevention of AD. PMID:25469702

  3. Ecological Evidence of Intensive Cultivation of Oaks by California Indians: Implications

    E-print Network

    Standiford, Richard B.

    . In the oak savannas of California the various species of native oaks occur as individual large trees on steeper slopes and in draws. Young trees are notably few or absent in oak savannas. The predominance of old oaks and a lack of regeneration means that the population of oaks in these savannas

  4. The increasing scarcity of red oaks in Mississippi River floodplain forests: Influence of the residual overstory

    Microsoft Academic Search

    Chadwick Dearing Oliver; E. C. Burkhardt; Daniel A. Skojac

    2005-01-01

    Red oaks – cherrybark oak (Quercus pagoda Raf.), willow oak (Quercus phellos L.), water oak (Quercus nigra L.), and Nuttall oak (Quercus texana Buckley; aka: Quercus nuttallii Palmer) – are not regrowing in Mississippi Delta river floodplain forests in the southeastern United States in sufficient numbers to sustain the former species composition and timber and wildlife values. Even if vigorous

  5. Associations Among Breeding Birds and Gambel Oak in Southwestern Ponderosa Pine Forests

    Microsoft Academic Search

    Stephanie Jentsch; R. William Mannan; Brett G. Dickson; William M. Block

    2008-01-01

    Ponderosa pine (Pinus ponderosa) forests with Gambel oak (Quercus gambelii) are associated with higher bird abundance and diversity than are ponderosa pine forests lacking Gambel oak. Little is known, however, about specific structural characteristics of Gambel oak trees, clumps, and stands that may be important to birds in ponderosa pine-Gambel oak (hereafter pine-oak) forests. We examined associations among breeding birds

  6. 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

  7. 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.

  8. Environmental restoration: Oak Ridge National Laboratory perspective

    SciTech Connect

    Garland, S.B. II.

    1991-01-01

    The Oak Ridge National Laboratory (ORNL), located in Oak Ridge, Tennessee, is a multipurpose research facility managed by Martin Marietta Energy Systems, Inc. (Energy Systems) for the US Deparment of Energy-Oak Ridge Operations (DOE). The operation of ORNL has resulted in a legacy of contaminated and potentially contaminated facilities, research areas, and waste management areas that may require remediation. The most recent inventory of remediation sites has identified approximately 400 individual sites that will require investigation and possibly remediation. The Remedial Action program (RAP) was established at ORNL in 1985 to conduct the investigations, studies,and remediation necessary to prevent unacceptable risks to the environment and to the public from this legacy of contaminated sites. Then, in 1989 a central Environmental Restoration program (ERP) was established that consolidates the previous RAPs at all five sites managed by Energy Systems for DOE. This paper describes how a program was developed to solve the large and diverse problems associated with the environmental restoration of the ORNL. 3 figs., 1 tab.

  9. Processing of Oak Ridge Mixed Waste Labpacks

    SciTech Connect

    Estes, C. H.; Franco, P.; Bisaria, A.

    2002-02-26

    The Oak Ridge Site Treatment Plan (STP) issued under a Tennessee Commissioner's Order includes a compliance milestone related to treatment of mixed waste labpacks on the Oak Ridge sites. The treatment plan was written and approved in Fiscal Year 1997. The plan involved approximately 1,100 labpacks and 7,400 on-the-shelf labpackable items stored at three Department of Energy (DOE) sites on the Oak Ridge Reservation (ORR). The labpacks and labpack items consist of liquids and solids with various chemical constituents and radiological concerns. The waste must be processed for shipment to a commercial hazardous waste treatment facility or treatment utilizing a Broad Spectrum mixed waste treatment contract. This paper will describe the labpack treatment plan that was developed as required by the Site Treatment Plan and the operations implemented to process the labpack waste. The paper will discuss the labpack inventory in the treatment plan, treatment and disposal options, processing strategies, project risk assessment, and current project status.

  10. Stability analysis of White Oak Dam

    SciTech Connect

    NONE

    1995-04-11

    White Oak Dam is located in the White Oak Creek watershed which provides the primary surface drainage for Oak Ridge National Laboratory. A stability analysis was made on the dam by Syed Ahmed in January 1994 which included an evaluation of the liquefaction potential of the embankment and foundation. This report evaluates the stability of the dam and includes comments on the report prepared by Ahmed. Slope stability analyses were performed on the dam and included cases for sudden drawdown, steady seepage, partial pool and earthquake. Results of the stability analyses indicate that the dam is stable and failure of the structure would not occur for the cases considered. The report prepared by Ahmed leads to the same conclusions as stated above. Review of the report finds that it is complete, well documented and conservative in its selection of soil parameters. The evaluation of the liquefaction potential is also complete and this report is in agreement with the findings that the dam and foundation are not susceptible to liquefaction.

  11. 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.

  12. Molecular cloning, characterization and expression analysis of cathepsin O in silkworm Bombyx mori related to bacterial response.

    PubMed

    Zhang, Kui; Su, Jingjing; Chen, Siyuan; Yu, Shuang; Tan, Juan; Xu, Man; Liang, Hanghua; Zhao, Yuzu; Chao, Huijuan; Yang, Liqun; Cui, Hongjuan

    2015-08-01

    Cathepsins are the main members of the cysteine family and play important roles in immune response in vertebrates. The Cathepsin O of Bombyx mori (BmCathepsin O) was cloned from the hemocytes by the rapid amplification of cDNA ends (RACE). The genomic DNA was 6131bp long with a total of six exons and five introns. Its pre-mRNA was spliced to generate two spliceosomes. By comparisons with other reported cathepsins O, it was concluded that the identity between them ranged from 29 to 39%. Expression analysis indicated that BmCathepsin O was specific-expressed in hemocytes, and highly expressed at the 4th molting and metamorphosis stages. Immunofluorescence assay and qRT-PCR showed that BmCathepsin O was expressed in granulocytes and plasmatocytes. Interestingly, BmCathepsin O was significantly up-regulated after stimulated by 20-hydroxyecdysone (20-E) in vivo, which suggested that BmCathepsin O may be regulated by 20E. Moreover, activation of BmCathepsin O was also observed in hemocytes challenged by Escherichia coli, indicating its potential involvement in the innate immune system of silkworm, B. mori. In summary, our studies provide a new insight into the functional features of Cathepsin O. PMID:25996894

  13. Genetic mapping of RAPD markers linked to the densonucleosis refractoriness gene, nsd-1, in the silkworm, Bombyx mori.

    PubMed

    Abe, H; Harada, T; Kanehara, M; Shimada, T; Ohbayashi, F; Oshiki, T

    1998-08-01

    In the silkworm, Bombyx mori, nonsusceptibility to B. mori densonucleosis virus type-1 (BmDNV-1) is controlled by a recessive gene, nsd-1 (nonsusceptibility to DNV-1), located on the twenty-first chromosome. We investigated genetic linkage between five random amplified polymorphic DNA (RAPD) markers and the +nsd-1 gene. Initially, we constructed the CSD-1 strain (nsd-1/+) which is congenic to strain C137 (nsd-1/nsd-1) for the twenty-first chromosome, starting with a female of C137 and a male of strain J137 (+nsd-1/+nsd-1). For the crossing over experiment, a female of C137 was crossed with a male (nsd-1/+) of CSD-1. Segregation analysis showed that the most closely linked RAPD marker mapped 3.0 cM distant from +nsd-1. A more specific marker for +nsd-1 was made by converting this RAPD band into a sequence characterized amplified region (SCAR) using a series of newly designed primer pairs based on its DNA sequence. PMID:9880921

  14. A novel TATA-box-binding factor from the silk glands of the mulberry silkworm, Bombyx mori.

    PubMed Central

    Srinivasan, Lakshmi; Gopinathan, Karumathil P

    2002-01-01

    The presence of one or more TATATAA motifs in the flanking sequences of individual members of a multi-gene tRNA(Gly)(1) family from the mulberry silkworm, Bombyx mori, negatively modulated the transcription of the gene copies. Characterization of proteins from posterior silk gland nuclear extracts, binding to the TATATAA motif, identified a novel 43 kD protein, designated here as P43 TATA-box-binding factor (TBF). The protein was purified to homogeneity. P43 TBF binding was highly sequence-specific and showed a 100-fold-higher affinity for binding than the TATA-box-binding protein (TBP). The protein also showed binding to the TATAAA sequence of the actin5C promoter. P43 TBF inhibited transcription of all the tRNA genes examined, as well as RNA polymerase II transcription from the actin5C promoter. The amino acid sequence of eleven peptides generated from P43 TBF did not share homology with proteins that bind the TATA box, such as TBP, TRF (TBP-related factor) or TLFs (TBP-like factors) reported from other sources. Inhibition of transcription of tRNA genes by P43 TBF could not be reversed by TBP. The inhibitory effect appeared to be exerted through sequestration of the associated transcription factors. PMID:11964150

  15. Effects of diet-deprivation and physical stimulation on the feeding behaviour of the larvae of the silkworm, Bombyx mori.

    PubMed

    Nagata, Shinji; Nagasawa, Hiromichi

    2006-08-01

    Continuous observations of larvae of the silkworm, Bombyx mori, revealed that feeding occurred at regular intervals throughout larval development. To investigate possible factors influencing meal-timing, the behaviours of diet-deprived Bombyx larvae were also analysed. Diet-deprivation resulted in longer durations of the first meals after diet replacement, but did not affect feeding patterns. Furthermore, long-term diet-deprivation promoted wandering behaviour and a consequent delay in feeding after diet replacement. Under diet-deprivation conditions, meal-starts appeared to be inducible by defecation and physical stimulation. However, stimulation-induced meal-starts were dependent on the time elapsed since the larvae's previous meals. Provided that more than 1h had elapsed since their previous meals, larvae could be induced to feed by defecation and tapping. At less than 1h post-meal, larvae were less likely to begin feeding after defecation or physical stimulation. Activated locomotions such as wandering and feeding were observed in the long-term diet-deprived larvae only after diet blocks were replaced, while long-term diet-deprived larvae did not show activated locomotion during the absence of diet blocks. Collectively, these data suggest that a combination of elevated locomotion activity and the presence of diet may be necessary for the initiation of feeding in diet-deprived larvae. PMID:16828111

  16. Mutation of a Cuticular Protein, BmorCPR2, Alters Larval Body Shape and Adaptability in Silkworm, Bombyx mori

    PubMed Central

    Qiao, Liang; Xiong, Gao; Wang, Ri-xin; He, Song-zhen; Chen, Jie; Tong, Xiao-ling; Hu, Hai; Li, Chun-lin; Gai, Ting-ting; Xin, Ya-qun; Liu, Xiao-fan; Chen, Bin; Xiang, Zhong-huai; Lu, Cheng; Dai, Fang-yin

    2014-01-01

    Cuticular proteins (CPs) are crucial components of the insect cuticle. Although numerous genes encoding cuticular proteins have been identified in known insect genomes to date, their functions in maintaining insect body shape and adaptability remain largely unknown. In the current study, positional cloning led to the identification of a gene encoding an RR1-type cuticular protein, BmorCPR2, highly expressed in larval chitin-rich tissues and at the mulberry leaf-eating stages, which is responsible for the silkworm stony mutant. In the Dazao-stony strain, the BmorCPR2 allele is a deletion mutation with significantly lower expression, compared to the wild-type Dazao strain. Dysfunctional BmorCPR2 in the stony mutant lost chitin binding ability, leading to reduced chitin content in larval cuticle, limitation of cuticle extension, abatement of cuticle tensile properties, and aberrant ratio between internodes and intersegmental folds. These variations induce a significant decrease in cuticle capacity to hold the growing internal organs in the larval development process, resulting in whole-body stiffness, tightness, and hardness, bulging intersegmental folds, and serious defects in larval adaptability. To our knowledge, this is the first study to report the corresponding phenotype of stony in insects caused by mutation of RR1-type cuticular protein. Our findings collectively shed light on the specific role of cuticular proteins in maintaining normal larval body shape and will aid in the development of pest control strategies for the management of Lepidoptera. PMID:24514903

  17. THE POPULATION BIOLOGY OF OAK GALL WASPS (HYMENOPTERA: CYNIPIDAE)

    Microsoft Academic Search

    Graham N. Stone; Karsten Schonrogge; Rachel J. Atkinson; David Bellido; Juli Pujade-Villar

    2002-01-01

    ? Abstract Oak gall wasps (Hymenoptera: Cynipidae, Cynipini) are characterized by possession of complex,cyclically parthenogenetic,life cycles and the ability to induce a wide,diversity of highly complex,species- and generation-specific galls on oaks and other Fagaceae. The galls support species-rich, closed communities of inquilines and parasitoids that have become,a model,system,in community,ecology. We review recent advances in the ecology of oak cynipids, with

  18. Role of glycoconjugates of 3-methyl-4-hydroxyoctanoic acid in the evolution of oak lactone in wine during oak maturation.

    PubMed

    Wilkinson, Kerry L; Prida, Andrei; Hayasaka, Yoji

    2013-05-01

    Oak lactone is a natural component of oak wood, but it also exists in glycoconjugate precursor forms. This study concerned the role of glycoconjugates of 3-methyl-4-hydroxyoctanoic acid, specifically a galloylglucoside, glucoside, and rutinoside, in the evolution of oak lactone during cooperage and maturation. The glycoconjugate profiles of 10 French oak samples were obtained by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) using stable isotope dilution analysis. The galloylglucoside was found to be the predominant glycoconjugate precursor and ranged in concentration from 110 to 354 ?g/g. Maturation trials indicated the galloylglucoside undergoes acid-catalyzed hydrolysis after extraction into wine; after 12 months of maturation, the glucoside was the most abundant precursor, present at between 2- and 11-fold higher concentrations than those observed for powdered oak. Thermal degradation of glycoconjugates was observed only when oak samples were heated at 200 °C for 30 min, demonstrating their thermal stability. PMID:23565604

  19. Characterization and phylogenetic relationships among microsporidia infecting silkworm, Bombyx mori, using inter simple sequence repeat (ISSR) and small subunit rRNA (SSU-rRNA) sequence analysis.

    PubMed

    Rao, S Nageswara; Nath, B Surendra; Saratchandra, B

    2005-06-01

    This study is the first report on the genetic characterization and relationships among different microsporidia infecting the silkworm, Bombyx mori, using inter simple sequence repeat PCR (ISSR-PCR) analysis. Six different microsporidians were distinguished through molecular DNA typing using ISSR-PCR. Thus, ISSR-PCR analysis can be a powerful tool to detect polymorphisms and identify microsporidians, which are difficult to study with microscopy because of their extremely small size. Of the 100 ISSR primers tested, only 28 primers had reproducibility and high polymorphism (93%). A total of 24 ISSR primers produced 55 unique genetic markers, which could be used to differentiate the microsporidians from each other. Among the 28 SSRs tested, the most abundant were (CA)n, (GA)n, and (GT)n repeats. The degree of band sharing was used to evaluate genetic similarity between different microsporidian isolates and to construct a phylogenetic tree using Jaccard's similarity coefficient. The results indicate that the DNA profiles based on ISSR markers can be used as diagnostic tools to identify different microsporidia with considerable accuracy. In addition, the small subunit ribosomal RNA (SSU-rRNA) sequence gene was amplified, cloned, and sequenced from each of the 6 microsporidian isolates. These sequences were compared with 20 other microsporidian SSU-rRNA sequences to develop a phylogenetic tree for the microsporidia isolated from the silkworms. This method was found to be useful in establishing the phylogenetic relationships among the different microsporidians isolated from silkworms. Of the 6 microsporidian isolates, NIK-1s revealed an SSU-rRNA gene sequence similar to Nosema bombycis, indicating that NIK-1s is similar to N. bombycis; the remaining 5 isolates, which differed from each other and from N. bombycis, were considered to be different variants belonging to the species N. bombycis. PMID:16121233

  20. 77 FR 56587 - Safety Zone, Atlantic Intracoastal Waterway; Oak Island, NC

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-13

    ...Intracoastal Waterway; Oak Island, NC AGENCY: Coast Guard...Intracoastal Waterway at Oak Island, [[Page 56588...Intracoastal Waterway, mile 311.8, at Oak Island, North Carolina. The...use only one of these three methods. FOR...

  1. Sudden oak death 'here to stay' Jeanne Wirka, resident biologist at the Bouverie Preserve in Glen Ellen, stands next to a live oak that fell after it developed

    E-print Network

    California at Berkeley, University of

    Sudden oak death 'here to stay' Jeanne Wirka, resident biologist at the Bouverie Preserve in Glen Ellen, stands next to a live oak that fell after it developed sudden oak death in the heavily wooded:10 p.m. As sudden oak death continues to ravage Sonoma County woodlands, a secluded creek near Glen

  2. A simple and accurate method for generating co-dominant markers: an application of conformation-sensitive gel electrophoresis to linkage analysis in the silkworm.

    PubMed

    Yasukochi, Y

    1999-06-01

    A simple and sensitive method for linkage analysis is described, which is based on conformation-sensitive gel electrophoresis (CSGE). Using urea-containing agarose gels or a commercially available polyacrylamide-derived matrix, 13 polymorphic markers were newly identified for known genes of the silkworm, Bombyx mori, which had been scored as monomorphic by PCR-RFLP analysis. This method for detecting polymorphisms is quite sensitive, and can be performed with inexpensive reagents and apparatus that is available in most molecular biology laboratories. PMID:10394917

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

    PubMed

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

    2015-02-01

    The fungal pathogen Beauveria bassiana causes serious economic losses in sericulture. Its origin is usually attributed to the release of B. bassiana insecticides against pine caterpillars (Dendrolimus punctuatus). In the present study, 488 B. bassiana isolates obtained from silkworm (Bombyx mori) collected from 13 Chinese provinces, and 327 B. bassiana isolates obtained from D. punctatus collected from 9 provinces, were analyzed for population genetic structure using the ISSR technique based on genetic distance. A UPGMA dendrogram clustered them into three independent clades: two B. mori clades and one D. punctatus clade. A 3-D principal component analysis further divided them into two completely independent host groups, revealing high host-specificity. This suggested that white muscardine occurring in B. mori populations throughout southern China was not caused by any B. bassiana strain either naturally prevailing in D. punctatus populations or by any strain artificially released as a fungal insecticide against D. punctatus. We further investigated the genetic differentiation coefficient Gst and gene flow between B. mori-pathogenic and D. punctatus-pathogenic B. bassiana isolates from across China and from five provinces inhabited by both B. mori and D. punctatus. The Gst value across China was computed as 0.410, while the values of the five provinces ranged from 0.508 to 0.689; all above 0.25, which is the threshold for significant genetic differentiation. This suggests that B. bassiana strains isolated from the two different hosts maintained their respective heredity without a convergent homogenization trend, and reduces the possibility that the host range of the caterpillar isolates could expand and enhance their virulence in B. mori. These findings indicate that the use of B. bassiana does not threaten the safety of sericulture. PMID:25541121

  4. 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...

  5. Comprehensive integrated planning: A process for the Oak Ridge Reservation, Oak Ridge, Tennessee

    SciTech Connect

    NONE

    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.

  6. Potential Effects of Sudden Oak Death on Birds in Coastal Oak Woodlands1

    Microsoft Academic Search

    Donald E. Winslow; William D. Tietje

    Tree pathogens can affect community composition and structure over wide areas. Phytophthora ramorum, cause of sudden oak death (SOD), occurs in the wild in California from Humboldt County to southernmost Monterey County. P. ramorum has killed many trees at some sites and may spread to affect near and distant forests. The pathogen has not yet been detected in San Luis

  7. Oak Ridge National Laboratory 5-1 5. Oak Ridge National Laboratory

    E-print Network

    Pennycook, Steve

    from, the work carried out in the days of the Manhattan Project. #12;Oak Ridge Reservation Annual Site (Bethel and Melton) and on Chestnut Ridge. ORNL was established in 1943 as part of the secret Manhattan Project to pioneer a method for producing and separating plutonium. During the 1950s and 1960s

  8. Programmed cell death: cytochemical and X-ray microanalytical characterization of calcium compartments in neuromuscular junctions during the normal breakdown of the intersegmental muscles in the giant silkmoth Antheraea polyphemus.

    PubMed

    Beaulaton, J

    1988-03-01

    Calcium stores were cytochemically demonstrated using a combined oxalate-pyroantimonate method in the neuromuscular junctions of the degenerating intersegmental muscles in the giant silkmoth Antheraea polyphemus. The elemental composition of punctate precipitates of the reaction product was determined by electron probe X-ray microanalysis of unstained thin sections by energy-dispersive spectrometry and wavelength-dispersive spectrometry. The wavelength-dispersive spectra collected over terminal axons demonstrate a significant calcium signal and a trace of antimony. During the rapid lytic phase of spontaneous muscle degeneration, the calcium punctate deposits were detected in presynaptic terminals in the following sites: the synaptic vesicles and the mitochondria. Calcium precipitates were also found in the dense bodies and the mitochondria encountered in the glial convolutions. No calcium deposit was seen in the synaptic clefts and intercellular spaces of the subsynaptic reticulum of type I and type II. A comparison of calcium to antimony ratios between the terminal axons and the sarcoplasmic lysosomes revealed highly significant differences (P less than 0.001). Such a variability of the calcium to antimony ratio may be related to different conditions of precipitation or antimony diffusion in the different cell compartments. It was concluded that such synaptic terminals do not appear damaged in spite of the muscle degeneration and presumably continue to perform vital functions while the muscles are no longer contractile 20 h after adult ecdysis. PMID:3410737

  9. Expression of the hIGF-I gene driven by the Fhx/P25 promoter in the silk glands of germline silkworm and transformed BmN cells.

    PubMed

    Li, Yanmei; Cao, Guangli; Wang, Yang; Xue, Renyu; Zhou, Wenlin; Gong, Chengliang

    2011-03-01

    The expression of the human insulin-like growth factor (hIGF-I) gene driven by the Fhx/P25 promoter in the silk glands of transgenic silkworms (Bombyx mori) and in transformed silkworm cells, was achieved using BmN cells transfected with a piggyBac vector, pigA3GFP-Fhx/P25-hIGF-ie-neo containing a neomycin-resistance gene (neo), a green fluorescent protein gene (gfp), an hIGF-I gene, and a helper plasmid containing the piggyBac transposase sequence under the control of the B. mori actin 3 (A3) promoter. We selected stably transformed BmN cells expressing hIGF-I using the antibiotic G418. The expression level of hIGF-I was about 450 pg in 3 × 10(6) cells, determined by ELISA. The piggyBac vector was transferred into the silkworm eggs using sperm-mediated gene transfer. The expression level of hIGF-I per gram fresh posterior silk glands of G4 transgenic silkworms was approx. 150 ng. PMID:21072564

  10. Oak Ridge Leadership Computing Facility Position Paper

    SciTech Connect

    Oral, H Sarp [ORNL] [ORNL; Hill, Jason J [ORNL] [ORNL; Thach, Kevin G [ORNL] [ORNL; Podhorszki, Norbert [ORNL] [ORNL; Klasky, Scott A [ORNL] [ORNL; Rogers, James H [ORNL] [ORNL; Shipman, Galen M [ORNL] [ORNL

    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.

  11. Crush Testing at Oak Ridge National Laboratory

    SciTech Connect

    Feldman, Matthew R [ORNL

    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.

  12. 1Oak Ridge National Laboratory Science & Technology Highlights

    E-print Network

    Pennycook, Steve

    1Oak Ridge National Laboratory Science & Technology Highlights Published by Oak Ridge National and Energy Assurance (EEA) Programs at ORNL have a record of success in transferring technologies, they are often preferred when proprietary information of the private company is used and that party hopes to gain

  13. California Oaks and Fire: A Review and Case Study1

    E-print Network

    Standiford, Richard B.

    investigation of the effects of fire on young valley oak trees indicates that saplings of this species, similar551 California Oaks and Fire: A Review and Case Study1 Katherine A. Holmes,2, 3, Kari E. Veblen,2, 3 Truman P. Young,2, 3 and Alison M. Berry2, 3 Abstract California has a fire-prone Mediterranean

  14. White Oak Creek embayment sediment retention structure design and construction

    Microsoft Academic Search

    S. D. Van Hoesen; B. L. Kimmell; D. G. Page; R. B. Wilkerson; G. R. Hudson; J. L. Kauschinger; M. Zocolla

    1994-01-01

    White Oak Creek is the major surface water drainage throughout 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 level of Cobalt 60 in near surface sediment. Other contaminants present in the sediment included: lead, mercury, chromium, and PCBs. In October 1990,

  15. Oak Ridge Reservation Annual Site environmental report summary for 1994

    SciTech Connect

    NONE

    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.

  16. OAK RIDGE NATIONAL LABORATORY U. S. DEPARTMENT OF ENERGY

    E-print Network

    OAK RIDGE NATIONAL LABORATORY U. S. DEPARTMENT OF ENERGY BES Peer Review 12/6-8/2006 CONCEPTUAL.O. Box 2008 Oak Ridge, Tennessee 37831-6285 managed by UT-Battelle, LLC for the U.S. DEPARTMENT OF ENERGY....................................................................................................................................... xiii Executive Summary

  17. Review article Genetic improvement of oaks in North America

    E-print Network

    Paris-Sud XI, Université de

    Review article Genetic improvement of oaks in North America KC Steiner School of Forest Resources contexts of oak tree improvement in North America are described briefly, and the methods, species orchard to operational plantations. Quercus /genetic improvement / North america / review Résumé &mdash

  18. A Hog under an Oak I. A. Krylov

    E-print Network

    Givental, Alexander

    . . -, -; , ; , , . « », : « , ».-- « », : « ; ; , ; : ».-- «!» : « , , ». , , , . A Hog under an Oak I. A. Krylov A Hog under a mighty Oak Had glutted tons of tasty acorns, then, supine and perish ­ don't you care?" ''Not in the least!'' The Hog raised up its head. "Why would the prospect make

  19. Utilization Implications for Hardwoods Susceptible to Sudden Oak Death1

    E-print Network

    Standiford, Richard B.

    Utilization Implications for Hardwoods Susceptible to Sudden Oak Death1 John R. Shelly2 The number of woody species succumbing to Sudden Oak Death (SOD) infections and the number of counties where and dying trees creates numerous handling and disposal problems. One possible solution is to develop markets

  20. Effects of Restoring Oak Savannas on Bird Communities and Populations

    Microsoft Academic Search

    JEFFREY D. BRAWN

    2006-01-01

    Efforts to restore and maintain oak savannas in North America, with emphasis on the use of pre- scribed fire, have become common. Little is known, however, about how restoration affects animal populations, especially those of birds. I compared the breeding densities, community structure, and reproductive success of birds in oak savannas maintained by prescribed fire (12 sites) with those in

  1. NOAA-OakRidgeExpandClimate ORNL,GECollaborateonHigh-

    E-print Network

    Climate Modeling The National Oceanic and Atmospheric Administration will provide $215 million to Oak #12;· NOAA-OakRidgeExpandClimate Modeling · ORNL,GECollaborateonHigh- Efficiency--followed by the delivery of a $1.4 billion project on time, on budget and on scope--is one of the truly great success

  2. Oak Ridge National Laboratory U.S. Department of Energy

    E-print Network

    Oak Ridge National Laboratory U.S. Department of Energy Dec-05 ARM Thumbnail Browser ­ A new way data streams. From the thumbnail view, users can directly access larger-scale data plots called "quick;Oak Ridge National Laboratory U.S. Department of Energy Dec-05 ARM Thumbnail Browser ­ A new way

  3. OAK RIDGE NATIONAL LABORATORY U. S. DEPARTMENT OF ENERGY

    E-print Network

    Deiterding, Ralf

    simulation of plates subjected to shock loading under water and in air Ralf Deiterding Oak Ridge National method · Reflection of a Mach 2.38 shock in nitrogen at 43o wedge · 2nd order MUSCL scheme with Roe MUSCL scheme with Van Leer FVS, dimensional splitting #12;10 OAK RIDGE NATIONAL L

  4. Emergence of the sudden oak death pathogen Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora ramorum, the pathogen responsible for massive sudden oak death of tanoak, coast live oak and Japanese larch in the United States and the United Kingdom, is the latest example of an emerging pathogen. This review documents the emergence of P. ramorum based on detailed, recent evolutionar...

  5. [Genetic analysis and gene mapping of two novel quail-like mutants from the silkworm (Bombyx mori)].

    PubMed

    Zhao, Qiaoling; Wang, Wenbo; Chen, Anli; Qiu, Zhiyong; Xia, Dingguo; Qian, Heying; Shen, Xingjia

    2014-04-01

    Two novel body marking mutants were discovered during silkworm (Bombyx mori) breeding. The mutants have no obvious eye-spots compared with normal marking (+) individuals, but their star spots and semilunar markings on dorsal sides are normal, and there are dots and lines with longitudinal wave markings on dorsal sides of the 6th to 7th abdominal segments which consist quail markings in between star spots and semilunar markings. The whole body markings are very similar to that of quail mutant (q); thus these mutants are named as quail-like mutants (q-l). Young larvae of one mutant are in brown color, and develop normally. Their cocoons are regular and uniform in size. Thus, this mutant is designated as brown quail-like (q-lb). Another mutant's larvae are in light purple skin; thus this mutant is named as purple quail-like (q-lp). They take little amount of mulberry leaves, and are weak and develop slowly and unevenly. Their larval bodies and cocoons are small. Genetic analysis revealed that both q-lb and q-lp were recessive genes, and they were allelic, with q-lb recessive to q-lp. These genes are different from quail mutant (q) and located on the chromosome 8 after tested by the morphological markers, P3(2), p(2), Ze(3), L(4), re(5), E(6), q(7), I-a(9), ms(12), ch(13), oa(14), cts(16), mln(18), msn(19), rb(21) and so(26) and SSR markers. PMID:24846982

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

    PubMed

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

    2012-02-01

    The cocoon shell of the silkworm Bombyx mori consists of silk fibroin fiber (70%) surrounded by a sericin layer made up of sericin (25%) and non-sericin (5%) components. The non-sericin component which consists of carbohydrate, salt, wax, flavonoids and derivatives is often overlooked in applied research into sericin and its hydrolysate. Here, sericin and non-sericin compounds were obtained from the sericin layer of five types of cocoon shell by means of degumming in water followed by extraction and separation in ethanol. These ethanol extracts were found to mainly contain flavonoids and free amino acids possessing scavenging activities of the 2,2-diphenyl -1-picrylhydrazyl (DPPH) free radical and inhibiting activities of tyrosinase, which were much greater than the corresponding activities of the purified sericin proteins. The extracts also strongly inhibited ?-glucosidase while the sericins had no such activity. In particular, the inhibitory activities of the ethanol extract of Daizo cocoons were much greater than those of the other cocoons. The IC(50) values of the Daizo cocoons for DPPH free radicals, tyrosinase, and ?-glucosidase were 170, 27, and 110 ?g mL(-1), respectively. The bioactivities of the non-sericin component were much higher than the activity of sericin alone. In addition, the in vivo test showed preliminarily that the administration of the non-sericin component had effectively resistant activity against streptozocin (STZ) oxidation and that of the purified sericin could also evidently decrease the induction ratio of diabetic mice induced by STZ. Therefore, ethanol extract protocols of the sericin layer of cocoon shells provide a novel stock which, together with sericin protein, has potential uses in functional food, biotechnological and medical applications. PMID:22101964

  7. Use of plant-derived protein hydrolysates for enhancing growth of Bombyx mori (silkworm) insect cells in suspension culture.

    PubMed

    Kwon, Mi Sun; Dojima, Takashi; Park, Enoch Y

    2005-08-01

    The successful suspension culture of the Bombyx mori (silkworm) cell lines Bm5 and BmN4 without FBS (fetal-bovine serum) was first realized in Sf-900 II SFM (serum-free medium) (Gibco BRL, Rockville, MD, U.S.A.) supplemented with a plant-derived protein hydrolysate. The addition of 0.5% HyPep 1510 (Difco Co., Detroit, MI, U.S.A.) and 10 mM glutamine to Sf-900 II SFM at 4 days of culture was found effective in increasing the cell concentration to 8.5x10(6) cells/ml. The replacement of medium with Sf-900 II SFM supplemented with 0.5% HyPep 1510 at 6 days of culture increased the cell concentration by 1.1x10(7) cells/ml. When Sf-900 II SFM was supplemented with 0.5% Hypep 1510, 16 days, which was half of the conventional adaptation time, was sufficient for the B. mori cell line to adapt to SFM and shear stress while maintaining a stable viability. The beta-galactosidase activity in Sf-900 II SFM supplemented with 0.5% Hypep 1510 was 4.9x10(3) units/ml, which was 2-fold higher than that of the FBS-supplemented medium. By SDS/PAGE, only the band corresponding to beta-galactosidase was detected in the sample from the media supplemented with plant-derived protein hydrolysates, while thick bands corresponding to proteins having lower molecular masses than beta-galactosidase were detected in samples from the FBS-supplemented media. These results suggest that plant-derived protein hydrolysates are promising FBS substitutes for enhancing the growth of B. mori cells and facilitating the purification of recombinant proteins produced by baculovirus infection. PMID:15527419

  8. Genetic analysis of scattered populations of the Indian eri silkworm, Samia cynthia ricini Donovan: Differentiation of subpopulations

    PubMed Central

    Pradeep, Appukuttannair R.; Jingade, Anuradha H.; Singh, Choba K.; Awasthi, Aravind K.; Kumar, Vikas; Rao, Guruprasad C.; Prakash, N.B. Vijaya

    2011-01-01

    Deforestation and exploitation has led to the fragmentation of habitats and scattering of populations of the economically important eri silkworm, Samia cynthia ricini, in north-east India. Genetic analysis of 15 eri populations, using ISSR markers, showed 98% inter-population, and 23% to 58% intra-population polymorphism. Nei’s genetic distance between populations increased significantly with altitude (R2 = 0.71) and geographic distance (R2 = 0.78). On the dendrogram, the lower and upper Assam populations were clustered separately, with intermediate grouping of those from Barpathar and Chuchuyimlang, consistent with geographical distribution. The Nei’s gene diversity index was 0.350 in total populations and 0.121 in subpopulations. The genetic differentiation estimate (Gst) was 0.276 among scattered populations. Neutrality tests showed deviation of 118 loci from Hardy-Weinberg equilibrium. The number of loci that deviated from neutrality increased with altitude (R2 = 0.63). Test of linkage disequilibrium showed greater contribution of variance among eri subpopulations to total variance. D’2IS exceeded D’2ST, showed significant contribution of random genetic drift to the increase in variance of disequilibrium in subpopulations. In the Lakhimpur population, the peripheral part was separated from the core by a genetic distance of 0.260. Patchy habitats promoted low genetic variability, high linkage disequilibrium and colonization by new subpopulations. Increased gene flow and habitat-area expansion are required to maintain higher genetic variability and conservation of the original S. c. ricini gene pool. PMID:21931526

  9. Recent progress in molecular genetic studies on the carotenoid transport system using cocoon-color mutants of the silkworm.

    PubMed

    Tsuchida, Kozo; Sakudoh, Takashi

    2015-04-15

    The existence of tissue-specific delivery for certain carotenoids is supported by genetic evidence from the silkworm Bombyx mori and the identification of cocoon color mutant genes, such as Yellow blood (Y), Yellow cocoon (C), and Flesh cocoon (F). Mutants with white cocoons are defective in one of the steps involved in transporting carotenoids from the midgut lumen to the middle silk gland via the hemolymph lipoprotein, lipophorin, and the different colored cocoons are caused by the accumulation of specific carotenoids into the middle silk gland. The Y gene encodes carotenoid-binding protein (CBP), which is expected to function as the cytosolic transporter of carotenoids across the enterocyte and epithelium of the middle silk gland. The C and F genes encode the C locus-associated membrane protein, which is homologous to a mammalian high-density lipoprotein receptor-2 (Cameo2) and scavenger receptor class B member 15 (SCRB15), respectively; these membrane proteins are expected to function as non-internalizing lipophorin receptors in the middle silk gland. Cameo2 and SCRB15 belong to the cluster determinant 36 (CD36) family, with Cameo2 exhibiting specificity not only for lutein, but also for zeaxanthin and astaxanthin, while SCRB15 seems to have specificity toward carotene substrates such as ?-carotene and ?-carotene. These findings suggest that Cameo2 and SCRB15 can discriminate the chemical structure of lutein and ?-carotene from circulating lipophorin during uptake. These data provide the first evidence that CD36 family proteins can discriminate individual carotenoid molecules in lipophorin. PMID:25579881

  10. 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 ...

  11. Not all oak gall wasps gall oaks: the description of Dryocosmus rileypokei, a new, apostate species of Cynipini from California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cynipini gall wasps are commonly known as oak gall wasps for their almost exclusive use of oak (Quercus spp.) as their host plant. Previously, only three of the nearly1000 species of Cynipini have been recorded from a host plant other than Quercus. These three species are known from western chinqu...

  12. Session G--Madrean Pine-Oak Forest Dynamics--Barton Madrean Pine-Oak Forest in Arizona

    E-print Network

    Standiford, Richard B.

    Session G--Madrean Pine-Oak Forest Dynamics--Barton Madrean Pine-Oak Forest in Arizona: Past Dynamics, Present Problems1 Andrew M. Barton2 Abstract This paper synthesizes research on presettlement, ME 04938. e-mail: Barton@maine.edu. USDA Forest Service Gen. Tech. Rep. PSW-GTR-189. 2008. 185 #12

  13. An aerial radiological survey of the Oak Ridge Reservation, Oak Ridge, Tennessee. Date of survey: April 1992

    SciTech Connect

    Maurer, R.J.

    1993-04-01

    An aerial radiological survey of the Oak Ridge Reservation (ORR) and surrounding area in Oak Ridge, Tennessee, was conducted during the period March 30 to April 14,1992. The purpose of the survey was to measure and document the terrestrial radiological environment of the Oak Ridge Reservation for use in environmental management programs and emergency response planning. The aerial survey was flown at an altitude of 150 feet (46 meters) along a series of parallel lines 250 feet (76 meters) apart and included X-10 (Oak Ridge National Laboratory), K-25 (former Gaseous Diffusion Plant), Y-12 (Weapons Production Plant), the Freels Bend Area and Oak Ridge Institute for Science and Education, the East Fork Poplar Creek (100-year floodplain extending from K-25 to Y-12), Elza Gate (former uranium ore storage site located in the city of Oak Ridge), Parcel A, the Clinch River (river banks extending from Melton Hill Dam to the city of Kingston), and the CSX Railroad Tracks (extending from Y-12 to the city of Oak Ridge). The survey encompassed approximately 55 square miles (1 41 square kilometers) of the Oak Ridge Reservation and surrounding area.

  14. White Oak Dam stability analysis. Volume I

    SciTech Connect

    Ahmed, S.B.

    1994-01-01

    A parametric study was conducted to evaluate the stability of the White Oak Dam (WOD) embankment and foundation. Slope stability analyses were performed for the upper and lower bound soil properties at three sections of the dam using the PCSTABL4 computer program. Minimum safety factors were calculated for the applicable seismic and static loading conditions. Liquefaction potential of the dam embankment and foundation solid during the seismic event was assessed by using simplified procedures. The WOD is classified as a low hazard facility and the Evaluation Basis Earthquake (EBE) is defined as an earthquake with a magnitude of m{sub b} = 5.6 and a Peak Ground Accelerator (PGA) of 0.13 g. This event is approximately equivalent to a Modified Mercalli Intensity of VI-VIII. The EBE is used to perform the seismic evaluation for slope stability and liquefaction potential. Results of the stability analyses and the liquefaction assessment lead to the conclusion that the White Oak Dam is safe and stable for the static and the seismic events defined in this study. Ogden Environmental, at the request of MMES, has checked and verified the calculations for the critical loading conditions and performed a peer review of this report. Ogden has determined that the WOD is stable under the defined static and seismic loading conditions and the embankment materials are in general not susceptible to liquefaction.

  15. The Oak Ridge Isochronous Cyclotron Refurbishment Project

    SciTech Connect

    Mendez, II, Anthony J [ORNL; Ball, James B [ORNL; Dowling, Darryl T [ORNL; Mosko, Sigmund W [ORNL; Tatum, B Alan [ORNL

    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.

  16. FITTING FIRE INTO THE OAK REGENERATION PROCESS: WHAT WORKS AND WHAT DOESN'T

    Microsoft Academic Search

    Patrick Brose; Thomas Schuler; Jeffrey Ward

    In the past decade, the use of prescribed fire has increased in the mixed-oak (Quercus spp.) forests of the eastern United States to help overcome the chronic lack of abundant, vigorous oak regeneration (Yaussy 2000). However, prescribed burns implemented under inappropriate circumstances can result in failure to establish oak regeneration and\\/or loss of existing oak reproduction. To correctly use fire

  17. Melton Valley Storage Tanks Capacity Increase Project, Oak Ridge National Laboratory, Oak Ridge, Tennessee

    SciTech Connect

    NONE

    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.

  18. 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.

  19. Oak Grove Fork Habitat Improvement Project, 1988 Annual Report.

    SciTech Connect

    Bettin, Scott (Mount Hood National Forest, Clackamas River Ranger District, Estacada, OR)

    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.

  20. 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.

  1. Accumulation of Glycoconjugates of 3-Methyl-4-hydroxyoctanoic Acid in Fruits, Leaves, and Shoots of Vitis vinifera cv. Monastrell following Foliar Applications of Oak Extract or Oak Lactone.

    PubMed

    Pardo-Garcia, Ana I; Wilkinson, Kerry L; Culbert, Julie A; Lloyd, Natoiya D R; Alonso, Gonzalo L; Salinas, M Rosario

    2015-05-13

    Grapevines are capable of absorbing volatile compounds present in the vineyard during the growing season, and in some cases, volatiles have been found to accumulate in fruits or leaves in glycoconjugate forms, that is, with one or more sugar moieties attached. The presence of oak lactone in wine is usually attributable to oak maturation, but oak lactone has been detected in wines made with fruit from grapevines treated with oak extract or oak lactone. This study investigated the accumulation of glycoconjugates of 3-methyl-4-hydroxyoctanoic acid (i.e., the ring-opened form of oak lactone) in the fruits, leaves, and shoots of Monastrell grapevines following foliar application of either oak extract or oak lactone at approximately 7 days postveraison. Fruits, leaves, and shoots were collected at three different time points, including at maturity. The oak lactone content of fruit was determined by gas chromatography-mass spectrometry, with declining concentrations observed in fruit from grapevines treated with oak lactone with ripening. The concentrations of a ?-d-glucopyranoside of 3-methyl-4-hydroxyoctanoic acid in fruits, leaves, and shoots was determined by liquid chromatography-tandem mass spectrometry, with the highest oak lactone glucoside levels observed in leaves of grapevines treated with oak lactone. A glucose-glucose disaccharide was also tentatively identified. These results demonstrate both ring-opening and glycosylation of oak lactone occurred after experimental treatments were imposed. PMID:25912091

  2. RIP-seq of BmAgo2-associated small RNAs reveal various types of small non-coding RNAs in the silkworm, Bombyx mori

    PubMed Central

    2013-01-01

    Background Small non-coding RNAs (ncRNAs) are important regulators of gene expression in eukaryotes. Previously, only microRNAs (miRNAs) and piRNAs have been identified in the silkworm, Bombyx mori. Furthermore, only ncRNAs (50-500nt) of intermediate size have been systematically identified in the silkworm. Results Here, we performed a systematic identification and analysis of small RNAs (18-50nt) associated with the Bombyx mori argonaute2 (BmAgo2) protein. Using RIP-seq, we identified various types of small ncRNAs associated with BmAGO2. These ncRNAs showed a multimodal length distribution, with three peaks at ~20nt, ~27nt and ~33nt, which included tRNA-, transposable element (TE)-, rRNA-, snoRNA- and snRNA-derived small RNAs as well as miRNAs and piRNAs. The tRNA-derived fragments (tRFs) were found at an extremely high abundance and accounted for 69.90% of the BmAgo2-associated small RNAs. Northern blotting confirmed that many tRFs were expressed or up-regulated only in the BmNPV-infected cells, implying that the tRFs play a prominent role by binding to BmAgo2 during BmNPV infection. Additional evidence suggested that there are potential cleavage sites on the D, anti-codon and T?C loops of the tRNAs. TE-derived small RNAs and piRNAs also accounted for a significant proportion of the BmAgo2-associated small RNAs, suggesting that BmAgo2 could be involved in the maintenance of genome stability by suppressing the activities of transposons guided by these small RNAs. Finally, Northern blotting was also used to confirm the Bombyx 5.8 s rRNA-derived small RNAs, demonstrating that various novel small RNAs exist in the silkworm. Conclusions Using an RIP-seq method in combination with Northern blotting, we identified various types of small RNAs associated with the BmAgo2 protein, including tRNA-, TE-, rRNA-, snoRNA- and snRNA-derived small RNAs as well as miRNAs and piRNAs. Our findings provide new clues for future functional studies of the role of small RNAs in insect development and evolution. PMID:24074203

  3. The human health impacts of the Oak processionary moth

    E-print Network

    The human health impacts of the Oak processionary moth Dr Olivier le Polain, Dr Barry Walsh #12 of the trees (30m): 47/69 (68%) - Symptoms: rash (100%), itchy eyes (29.7%), breathing problems (4.2%) #12

  4. VIEW ALONG SEVENTEENTH STREET. NOTE THE MATURE SILK OAK TREES ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    VIEW ALONG SEVENTEENTH STREET. NOTE THE MATURE SILK OAK TREES LINING THE STREET, WHICH DO NOT PROVIDE A CANOPY VIEW FACING NORTHWEST. - Hickam Field, Hickam Historic Housing, Honolulu, Honolulu County, HI

  5. 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.

  6. 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

  7. Name: VIRGINIA S. HINSHAW Place of Birth: Oak Ridge, Tennessee

    E-print Network

    Name: VIRGINIA S. HINSHAW Place of Birth: Oak Ridge, Tennessee Citizenship: U.S.A. Office Address of Virginia, Richmond, Virginia 1974­75 Research Virologist, Office of Environmental Research, University

  8. Name: VIRGINIA S. HINSHAW Place of Birth: Oak Ridge, Tennessee

    E-print Network

    Name: VIRGINIA S. HINSHAW Place of Birth: Oak Ridge, Tennessee Citizenship: U.S.A. Office Address of Virginia, Richmond, Virginia 1974-75 Research Virologist, Office of Environmental Research, University

  9. Range Vegetation Response to Burning Thicketized Live Oak Savannah.

    E-print Network

    Scifres, C.J.; Kelly, D.M.

    1979-01-01

    , yellow indiangrass, gulfdune paspalum, and Dichanthelium species. Major forb species are western ragweed, plains wild- indigo, Texas croton, hairy bonamia, and Cassia specles. The major woody species is live oak, which occurs as scattered large trees...

  10. Oak seedling growth and ectomycorrhizal colonization are less in eastern hemlock stands infested with hemlock woolly adelgid than in adjacent oak stands.

    PubMed

    Lewis, James D; Licitra, Jeff; Tuininga, Amy R; Sirulnik, Abby; Turner, Gregory D; Johnson, Jacqui

    2008-04-01

    Invasive, non-indigenous, phytophagous insects have caused widespread declines in several dominant tree species. The decline in dominant tree species may lead to cascading effects on other tree and microbial species and their interactions, affecting forest recovery following the decline. In the eastern USA, eastern hemlock (Tsuga canadensis (L.) Carr) is declining because of infestation by the hemlock woolly adelgid (HWA; Adelges tsugae Annand). Northern red oak (Quercus rubra L.) is a common replacement species in declining hemlock stands, but reduced mycorrhizal inoculum potential in infested hemlock stands may cause oak to grow more slowly compared with oak in oak stands. We grew red oak seedlings for one growing season in declining hemlock-dominated stands infested with HWA and in adjacent oak-dominated stands. Ectomycorrhizal root tip density and morphotype richness in soil cores were 63 and 27% less, respectively, in declining hemlock stands than in oak stands. Similarly, ectomycorrhizal percent colonization and morphotype richness on oak seedlings were 33 and 30% less, respectively, in declining hemlock stands than in oak stands. In addition, oak seedlings in declining hemlock stands had 29% less dry mass than oak seedlings in oak stands. Analysis of covariance indicated that morphotype richness could account for differences in oak seedling dry mass between declining hemlock stands and oak stands. Additionally, oak seedling dry mass in declining hemlock stands significantly decreased with decreasing ectomycorrhizal percent colonization and morphotype richness. These results suggest that oak seedling growth in declining hemlock stands is affected by reduced ectomycorrhizal inoculum potential. Further, the rate of forest recovery following hemlock decline associated with HWA infestation may be slowed by indirect effects of HWA on the growth of replacement species, through effects on ectomycorrhizal colonization and morphotype richness. PMID:18244948

  11. Water quality monitoring report for the White Oak Creek Embayment

    Microsoft Academic Search

    C. J. Ford; M. T. Wefer

    1993-01-01

    Water quality monitoring activities that focused on the detection of resuspended sediments in the Clinch River were conducted in conjunction with the White Oak Creek Embayment (WOCE) time-critical Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) removal action to construct a sediment-retention structure at the mouth of White Oak Creek (WOC). Samples were collected by use of a 24-h composite

  12. A Comparison of Leaf Physiology and Anatomy of Two Himalayan Oaks in Response to Different Light Environments

    Microsoft Academic Search

    Rajesh Thadani; Graeme P. Berlyn; Mark S. Ashton

    2009-01-01

    Leaf physiological and anatomical attributes of seedlings of two Himalayan oaks grown in different light conditions were examined. Both species are stand forming with banj oak (Quercus leucotrichophora) dominating at elevations of 1400–2300 m whereas tilonj oak (Quercus floribunda) is common from 2100–2700 m altitudes. Both oaks are hypostomatous and have sclerophyllous leaves. While Q. leucotrichophora oak leaves were larger,

  13. Potential mode of protection of silkworm pupae from environmental stress by harboring the bacterial biofilm on the surfaces of silk cocoons.

    PubMed

    Halder, Pranab K; Naskar, Deboki; Kumar, Akash; Yao, Juming; Kundu, Subhas C; Ghosh, Anindya S

    2015-02-01

    The silkworm forms cocoon to protect its pupa that survives for months inside the cocoon without being affected by various environmental stresses. To understand the possible mode of pupal survival within the cocoon encasement, we investigate the cause that protects the cocoon. During the end of the spinning process, we have isolated different bacterial species from the cocoon surface. These are identified using molecular techniques and checked for their abilities to form biofilm in vitro. The bacteria are able to form biofilm either individually or in consortia. Of which, Bacillus and Erwinia species are prominent biofilm formers. Interestingly, these bacteria have the ability to form biofilm on the cocoon mimetic surface of the silk protein Sericin Hope that contains only sericin. The origin and the behavior of the bacteria lead us to hypothesize the possible role of biofilm layer on the cocoon surface, which provides protection from adverse environmental conditions. PMID:25292249

  14. Identification and molecular characterization of a sex chromosome rearrangement causing a soft and pliable (spli) larval body phenotype in the silkworm, Bombyx mori.

    PubMed

    Fujii, Tsuguru; Kuwazaki, Seigo; Yamamoto, Kimiko; Abe, Hiroaki; Ohnuma, Akio; Katsuma, Susumu; Mita, Kazuei; Shimada, Toru

    2010-01-01

    We carried out genetic and cytogenetic analyses of X-ray-induced deleterious Z chromosomes that result in a soft and pliable (spli) phenotype in the silkworm, Bombyx mori. In a B. mori strain with a spli phenotype, we found the Z chromosome broken between the sch (1-21.5) and od (1-49.6) loci. We also found a chromosomal fragment bearing a fifth-chromosome locus for egg and eye pigmentation fused to a Z chromosome fragment. By means of fluorescence in situ hybridization using bacterial artificial chromosome clones as probes, we confirmed that the fused chromosome is composed of a fragment of chromosome 5 and a fragment of the Z chromosome. Moreover, a predicted gene, GA002017, the Bombyx ortholog of the Drosophila gene acj6 (Bmacj6), was completely deleted by the Z chromosome breakage event. The relationship between Bmacj6 and the spli phenotype is discussed. PMID:20130748

  15. 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.

  16. Influence of climate on tree rings and vessel features in red oak and white oak growing near their northern distribution limit, southwestern Quebec, Canada

    Microsoft Academic Search

    J. C. Tardif; F. Conciatori

    2006-01-01

    Little is known about environmental controls on vessel features in ring-porous tree species. Our objectives were to assess (i) the association between tree-ring descriptors (vessels and width) and climate in two oak species, white oak, Quercus alba L., and red oak, Quercus rubra L., and (ii) the utility of vessel series in climate reconstruc- tion. The study was conducted in

  17. Vegetation and Small Vertebrates of Oak Woodlands at Low and High Risk for Sudden Oak Death in San Luis Obispo County, California1

    Microsoft Academic Search

    Douglas J. Tempel; William D. Tietje; Donald E. Winslow

    San Luis Obispo County contains oak woodlands at varying levels of risk of sudden oak death (SOD), caused by a fungal pathogen (Phytophthora ramorum) that in the past decade has killed thousands of oak (Quercus spp.) and tanoak (Lithocarpus densiflorus) trees in California. SOD was most recently detected 16 km north of the San Luis Obispo County line. Low-risk woodlands

  18. The proceedings of the sudden oak death second science symposium: the state of our knowledge The Effects of Sudden Oak Death on

    E-print Network

    Standiford, Richard B.

    The proceedings of the sudden oak death second science symposium: the state of our knowledge 559 The Effects of Sudden Oak Death on Wildlife-Can Anything Be Learned From the American Chestnut Blight?1 area. In 1995, Sudden Oak Death (SOD) was identified near Mill Valley, California. Caused by the fungus

  19. RCRA Facility Investigation report for Waste Area Grouping 6 at Oak Ridge National Laboratory, Oak Ridge, Tennessee

    SciTech Connect

    Wilson, R. C.; Lewis, K. K.

    1991-09-01

    This report presents data and information related to remedial investigation studies for Oak Ridge National Laboratory (ORNL). Information is included on a soil gas survey, surface radiological investigations of waste areas, and well installation for ground water monitoring. (CBS)

  20. Consumption of Bt Rice Pollen Containing Cry1C or Cry2A Protein Poses a Low to Negligible Risk to the Silkworm Bombyx mori (Lepidoptera: Bombyxidae)

    PubMed Central

    Yang, Yan; Liu, Yue; Cao, Fengqin; Chen, Xiuping; Cheng, Lisheng; Romeis, Jörg; Li, Yunhe; Peng, Yufa

    2014-01-01

    By consuming mulberry leaves covered with pollen from nearby genetically engineered, insect-resistant rice lines producing Cry proteins derived from Bacillus thuringiensis (Bt), larvae of the domestic silkworm, Bombyx mori (Linnaeus) (Lepidoptera: Bombyxidae), could be exposed to insecticidal proteins. Laboratory experiments were conducted to assess the potential effects of Cry1C- or Cry2A-producing transgenic rice (T1C-19, T2A-1) pollen on B. mori fitness. In a short-term assay, B. mori larvae were fed mulberry leaves covered with different densities of pollen from Bt rice lines or their corresponding near isoline (control) for the first 3 d and then were fed mulberry leaves without pollen. No effect was detected on any life table parameter, even at 1800 pollen grains/cm2 leaf, which is much higher than the mean natural density of rice pollen on leaves of mulberry trees near paddy fields. In a long-term assay, the larvae were fed Bt and control pollen in the same way but for their entire larval stage (approximately 27 d). Bt pollen densities ?150 grains/cm2 leaf reduced 14-d larval weight, increased larval development time, and reduced adult eclosion rate. ELISA analyses showed that 72.6% of the Cry protein was still detected in the pollen grains excreted with the feces. The low exposure of silkworm larvae to Cry proteins when feeding Bt rice pollen may be the explanation for the relatively low toxicity detected in the current study. Although the results demonstrate that B. mori larvae are sensitive to Cry1C and Cry2A proteins, the exposure levels that harmed the larvae in the current study are far greater than natural exposure levels. We therefore conclude that consumption of Bt rice pollen will pose a low to negligible risk to B. mori. PMID:25014054