Science.gov

Sample records for mulberry wild silkworm

  1. Comparing the rheology of mulberry and "wild" silkworm spinning dopes.

    PubMed

    Holland, C; Porter, D; Vollrath, F

    2012-06-01

    Lepidoperan silks provide a superb opportunity for comparative studies of spinning and fiber characteristics. Comparing the four species, Bombyx mori (China), Actias selene (India), Antheraea yamamai (Japan), Gonometa postica (Africa), allows us to examine differences on the family, species, and race levels. Measured rheological properties were consistent with phylogenetic relationships and in the context of resource allocation and gland morphology. We propose that the thorough domestication of the mulberry silkworm B. mori for high silk yield has resulted in a compensatory optimization for spinning efficiency. This is in stark contrast to the wild silkworms, where Saturnids appear to minimize their energetic input toward silk output and G. postica seems to balance both. We conclude that comparative studies provide valuable baseline information for future biomimetic applications and modeling, as well as illuminating biologically important details of silk processing. PMID:22161905

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

    SciTech Connect

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

    1980-01-01

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

  3. [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

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

    PubMed

    Zhou, Lingyun; Zhao, Ye; Wang, Shuifeng

    2015-11-01

    Phytoremediation has been proven to be an environmentally sound alternative for the recovery of contaminated soils, and the economic profit that comes along with the process might stimulate its field use. This study investigated cadmium (Cd) transfer and detoxification mechanisms in a soil-mulberry-silkworm system to estimate the suitability of the mulberry and silkworm as an alternative method for the remediation of Cd-polluted soil; it also explored the underlying mechanisms regulating the trophic transfer of Cd. The results show that both the mulberry and silkworm have high Cd tolerance. The transfer factor suggests that the mulberry has high potential for Cd extraction from polluted soil. The subcellular distribution and chemical forms of Cd in mulberry leaves show that cell wall deposition and vacuolar compartmentalization play important role in Cd tolerance. In the presence of increasing Cd concentrations in silkworm food, detoxification mechanisms (excretion and homeostasis) were activated so that excess Cd was excreted in fecal balls, and metallothionein levels in the mid-gut, the posterior of the silk gland, and the fat body of silkworms were enhanced. And, the Cd concentrations in silk are at a low level, ranging from 0.02 to 0.21 mg kg(-1). Therefore, these mechanisms of detoxification can regulate Cd trophic transfer, and mulberry planting and silkworm breeding has high phytoremediation potential for Cd-contaminated soil. PMID:26169822

  5. Screening strains of the mulberry silkworm, Bombyx mori, for thermotolerance.

    PubMed

    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 BO(2) 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

  6. 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

  7. Spermidine enhances the silk production by mulberry silkworm.

    PubMed

    Lattala, Gayatri Manogna; Kandukuru, Kasturaiah; Gangupantula, Shamitha; Mamillapalli, Anitha

    2014-01-01

    Polyamines are ubiquitous low molecular weight polycationic aliphatic amines involved in diverse cellular processes. Spermidine (Spd), a polyamine, has been proved to be crucial for cell survival in various organisms. Our study reports the effect of Spd on the growth of Bombyx mori. Silkworms showed improved silk gland weight and economic parameters in the fifth instar larval stage when treated with different concentrations of Spd, in the range of 25-75 µM. The worms treated with Spd produced 31% more silk when compared with the control worms. Altogether, this study establishes that Spd-treated leaves can be fed into the larvae for better silk production. PMID:25502041

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

    PubMed

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

    2015-02-01

    Degradation behavior is very important in the field of silk-based biomaterials. Mulberry and nonmulberry silk fibroins are structurally and functionally distinguishable; however, no studies have examined the differences in the degradation behaviors of silk materials from various silkworm species. In this study, Ca(NO3)2 was used as a uniform solvent to obtain regenerated mulberry and nonmulberry (Antheraea pernyi and Antheraea yamamai) silk fibroin (SF) solutions, and the degradation behaviors of various SF scaffolds were examined. In vitro and in vivo results demonstrated that regenerated mulberry SF scaffolds exhibited significantly higher mass loss and free amino acid content release than did nonmulberry SF scaffolds. The differences in the primary structures and condensed structures between mulberry and nonmulberry SF contributed to the significant difference in degradation rates, in which the characteristic (-Ala-)n repeats, compact crystal structure and high α-helix and β-sheet contents make nonmulberry SF more resistant than mulberry SF to enzymatic degradation. Moreover, the Antheraea pernyi and Antheraea yamamai SFs possess similar primary structures and condensed structures, although a slight difference in degradation was observed; this difference might depend on the differences in molecular weight following the regeneration process. The results indicate that the original sources of SF significantly influence the degradation rates of SF-based materials; therefore, the original sources of SF should be fully considered for preparing tissue engineering scaffolds with matched degradation rates. PMID:25532470

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

    PubMed Central

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

    2015-01-01

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

  10. Eri silkworm (Samia ricini), a non-mulberry host system for AcMNPV mediated expression of recombinant proteins.

    PubMed

    Hosamani, Madhusudan; Basagoudanavar, Suresh H; Sreenivasa, B P; Inumaru, Shigeki; Ballal, Chandish R; Venkataramanan, Ramamurthy

    2015-12-20

    The baculovirus expression system (BVES) based on Autographa californica nucleopolyhedrovirus (AcMNPV) is widely used for the expression of eukaryotic proteins. Several insect cells/larvae that are permissive to AcMNPV have been routinely used as hosts to express heterologous proteins. Domesticated Eri silkworm (Samia ricini), reared in many parts of India, Japan and China, is a non-mulberry silkworm. The present study shows that the Eri silkworm larvae are susceptible to intra-haemocoelical inoculation of AcMNPV. The virus replicates in the larva, as indicated by an increased viral loads in the haemolymph upon injection of a recombinant AcMNPV carrying green fluorescent protein gene. The virus showed localized replication in different tissues including the fat body, haemocytes, tracheal matrix and in the Malphigian tubules. The larval system was successfully used to express heterologous protein, by infecting with a recombinant AcMNPV carrying the 3ABC coding sequence of foot-and-mouth disease virus (FMDV). The study shows that the Eri silkworm larva can be a potential alternative bioreactor, for scaling up of the recombinant proteins employing the baculovirus system. PMID:26467714

  11. 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

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

    PubMed

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

    2016-01-01

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

  13. MorusDB: a resource for mulberry genomics and genome biology.

    PubMed

    Li, Tian; Qi, Xiwu; Zeng, Qiwei; Xiang, Zhonghuai; He, Ningjia

    2014-01-01

    Mulberry is an important cultivated plant that has received the attention of biologists interested in sericulture and plant-insect interaction. Morus notabilis, a wild mulberry species with a minimal chromosome number is an ideal material for whole-genome sequencing and assembly. The genome and transcriptome of M. notabilis were sequenced and analyzed. In this article, a web-based and open-access database, the Morus Genome Database (MorusDB), was developed to enable easy-to-access and data mining. The MorusDB provides an integrated data source and an easy accession of mulberry large-scale genomic sequencing and assembly, predicted genes and functional annotations, expressed sequence tags (ESTs), transposable elements (TEs), Gene Ontology (GO) terms, horizontal gene transfers between mulberry and silkworm and ortholog and paralog groups. Transcriptome sequencing data for M. notabilis root, leaf, bark, winter bud and male flower can also be searched and downloaded. Furthermore, MorusDB provides an analytical workbench with some built-in tools and pipelines, such as BLAST, Search GO, Mulberry GO and Mulberry GBrowse, to facilitate genomic studies and comparative genomics. The MorusDB provides important genomic resources for scientists working with mulberry and other Moraceae species, which include many important fruit crops. Designed as a basic platform and accompanied by the SilkDB, MorusDB strives to be a comprehensive platform for the silkworm-mulberry interaction studies. Database URL: http://morus.swu.edu.cn/morusdb. PMID:24923822

  14. 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

  15. MorusDB: a resource for mulberry genomics and genome biology

    PubMed Central

    Li, Tian; Qi, Xiwu; Zeng, Qiwei; Xiang, Zhonghuai; He, Ningjia

    2014-01-01

    Mulberry is an important cultivated plant that has received the attention of biologists interested in sericulture and plant–insect interaction. Morus notabilis, a wild mulberry species with a minimal chromosome number is an ideal material for whole-genome sequencing and assembly. The genome and transcriptome of M. notabilis were sequenced and analyzed. In this article, a web-based and open-access database, the Morus Genome Database (MorusDB), was developed to enable easy-to-access and data mining. The MorusDB provides an integrated data source and an easy accession of mulberry large-scale genomic sequencing and assembly, predicted genes and functional annotations, expressed sequence tags (ESTs), transposable elements (TEs), Gene Ontology (GO) terms, horizontal gene transfers between mulberry and silkworm and ortholog and paralog groups. Transcriptome sequencing data for M. notabilis root, leaf, bark, winter bud and male flower can also be searched and downloaded. Furthermore, MorusDB provides an analytical workbench with some built-in tools and pipelines, such as BLAST, Search GO, Mulberry GO and Mulberry GBrowse, to facilitate genomic studies and comparative genomics. The MorusDB provides important genomic resources for scientists working with mulberry and other Moraceae species, which include many important fruit crops. Designed as a basic platform and accompanied by the SilkDB, MorusDB strives to be a comprehensive platform for the silkworm–mulberry interaction studies. Database URL: http://morus.swu.edu.cn/morusdb. PMID:24923822

  16. Phenotypic comparisons between wild relatives and cultivars of kiwifruit, persimmon, mulberry, and olive at the National Clonal Germplasm Repository, Davis, CA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phenotypic traits were characterized for 23 wild species and 4 cultivars of 4 clonal fruit crops including, Kiwifruit (Actinidia), Persimmon (Diospyros), Mulberry (Morus) and Olive (Olea). Across all four crops, the wild species varied distinctly, especially when compared with the cultivars. The wil...

  17. 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

  18. Comparative analysis of the integument transcriptomes of the black dilute mutant and the wild-type silkworm Bombyx mori.

    PubMed

    Wu, Songyuan; Tong, Xiaoling; Peng, Chenxing; Xiong, Gao; Lu, Kunpeng; Hu, Hai; Tan, Duan; Li, Chunlin; Han, Minjin; Lu, Cheng; Dai, Fangyin

    2016-01-01

    The insect cuticle is a critical protective shell that is composed predominantly of chitin and various cuticular proteins and pigments. Indeed, insects often change their surface pigment patterns in response to selective pressures, such as threats from predators, sexual selection and environmental changes. However, the molecular mechanisms underlying the construction of the epidermis and its pigmentation patterns are not fully understood. Among Lepidoptera, the silkworm is a favorable model for color pattern research. The black dilute (bd) mutant of silkworm is the result of a spontaneous mutation; the larval body color is notably melanized. We performed integument transcriptome sequencing of the wild-type strain Dazao and the mutant strains +/bd and bd/bd. In these experiments, during an early stage of the fourth molt, a stage at which approximately 51% of genes were expressed genome wide (RPKM ≥1) in each strain. A total of 254 novel transcripts were characterized using Cuffcompare and BLAST analyses. Comparison of the transcriptome data revealed 28 differentially expressed genes (DEGs) that may contribute to bd larval melanism, including 15 cuticular protein genes that were remarkably highly expressed in the bd/bd mutant. We suggest that these significantly up-regulated cuticular proteins may promote melanism in silkworm larvae. PMID:27193628

  19. Draft genome sequence of the mulberry tree Morus notabilis

    PubMed Central

    He, Ningjia; Zhang, Chi; Qi, Xiwu; Zhao, Shancen; Tao, Yong; Yang, Guojun; Lee, Tae-Ho; Wang, Xiyin; Cai, Qingle; Li, Dong; Lu, Mengzhu; Liao, Sentai; Luo, Guoqing; He, Rongjun; Tan, Xu; Xu, Yunmin; Li, Tian; Zhao, Aichun; Jia, Ling; Fu, Qiang; Zeng, Qiwei; Gao, Chuan; Ma, Bi; Liang, Jiubo; Wang, Xiling; Shang, Jingzhe; Song, Penghua; Wu, Haiyang; Fan, Li; Wang, Qing; Shuai, Qin; Zhu, Juanjuan; Wei, Congjin; Zhu-Salzman, Keyan; Jin, Dianchuan; Wang, Jinpeng; Liu, Tao; Yu, Maode; Tang, Cuiming; Wang, Zhenjiang; Dai, Fanwei; Chen, Jiafei; Liu, Yan; Zhao, Shutang; Lin, Tianbao; Zhang, Shougong; Wang, Junyi; Wang, Jian; Yang, Huanming; Yang, Guangwei; Wang, Jun; Paterson, Andrew H.; Xia, Qingyou; Ji, Dongfeng; Xiang, Zhonghuai

    2013-01-01

    Human utilization of the mulberry–silkworm interaction started at least 5,000 years ago and greatly influenced world history through the Silk Road. Complementing the silkworm genome sequence, here we describe the genome of a mulberry species Morus notabilis. In the 330-Mb genome assembly, we identify 128 Mb of repetitive sequences and 29,338 genes, 60.8% of which are supported by transcriptome sequencing. Mulberry gene sequences appear to evolve ~3 times faster than other Rosales, perhaps facilitating the species’ spread worldwide. The mulberry tree is among a few eudicots but several Rosales that have not preserved genome duplications in more than 100 million years; however, a neopolyploid series found in the mulberry tree and several others suggest that new duplications may confer benefits. Five predicted mulberry miRNAs are found in the haemolymph and silk glands of the silkworm, suggesting interactions at molecular levels in the plant–herbivore relationship. The identification and analyses of mulberry genes involved in diversifying selection, resistance and protease inhibitor expressed in the laticifers will accelerate the improvement of mulberry plants. PMID:24048436

  20. Identification of novel random amplified polymorphic DNAs (RAPDs) on the W chromosome of the domesticated silkworm, Bombyx mori, and the wild silkworm, B. mandarina, and their retrotransposable element-related nucleotide sequences.

    PubMed

    Abe, H; Kanehara, M; Terada, T; Ohbayashi, F; Shimada, T; Kawai, S; Suzuki, M; Sugasaki, T; Oshiki, T

    1998-08-01

    Genomic DNAs were compared between males and females of the domesticated silkworm, Bombyx mori, strains C108, C137, J137, p50, and WILD-W (constructed by crossing a wild silkworm, B. mandarina, female with a male of strain C108) by polymerase chain reaction (PCR) with 700 arbitrary 10-mer primers. Four female-specific RAPDs (W-Kabuki, W-Samurai, W-Kamikaze, and W-Yamato) were found. The sex chromosome formulas of B. mori and B. mandarina are ZW (XY) for the female and ZZ (XX) for the male. The four female-specific RAPDs are assumed to be derived from the W chromosome because the other chromosomes are shared by both sexes. A computer search for deduced amino acid sequences of these four RAPDs revealed that all of them showed homology to previously reported amino acid sequences encoded in known retrotransposable elements from various organisms. PMID:9880922

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

    NASA Astrophysics Data System (ADS)

    Yu, XiaoHui; Liu, Hong; Tong, Ling

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

  2. A novel splice variant of the decapentaplegic (dpp) gene in the wild silkworm, Bombyx mandarina.

    PubMed

    Kwak, Woori; Choi, Jung-Won; Ryul Kim, Seong; Choi, Kwang-Ho; Kim, Kee-Young; Goo, Tae-Won; Park, Seung-Won

    2015-10-23

    Decapentaplegic (dpp) is a member of the transforming growth factor-β superfamily. Although the dpp gene and related pathways are known to play important roles in insect development, few studies have examined its function in Bombyx mori and Bombyx mandarina. To date, there have been no previous reports on novel splice variants of dpp in silkworm. In the present study, we conducted RT-PCR to examine dpp expression in the mid-gut tissue of B. mandarina and discovered a novel dpp isoform. The isoform sequence was confirmed using sequencing analysis and found to have 333 bp deletion compared to full-length cDNA encoding dpp. The deleted sequence encodes a region of the latency associated peptide (LAP) region of transforming growth factor-β (TGF-β), which may affect the activity and specificity of TGF-β. Using variant calling analyses, we detected 7 candidate single nucleotide variants (SNVs) for different alternative splicing in dpp. This is the first report of a novel splice variant of the dpp gene in B. mandarina and these results provide insight about the domestication process and distinct phenotypic traits of B. mori and B. mandarina. PMID:26365352

  3. Complete mitochondrial genome of the wild eri silkworm, Samia canningi (Lepidoptera: Saturniidae).

    PubMed

    Shantibala, T; Victor, Th; Luikham, Reeta; Arunkumar, K P; Sharma, H Debaraj; Lokeshwari, R K; Kim, Iksoo

    2016-01-01

    The saturniid silkworm species of the genus Samia are potential silk producing insects. Thus, Samia canningi (Lepidoptera: Saturniidae) is a potential candidate to introduce for silk production. The complete mitochondrial genome of S. canningi was 15,384 bp long that contained 37 genes along with a control region. The arrangement of the PCGs was same as the majority of Lepidoptera, presenting the order, trnM/trnI/trnQ between nad2 and control region. Twelve of 13 PCGs started with ATN codons, but cox2 with GTG, which is often found in insects. Genes overlapped in a total of 29 bp, 221 bp of intergenic spacer sequences was found in seventeen regions and the longest 54 bp one was found between trnQ and nad2 as typical in Lepidoptera. In lrRNA, the 21-bp long, tandemly duplicated repeat was characteristically found (TAAAATTATTTATAATATAAA) between 13,663 and 13,706. AT rich region has the motif "ATAGA" and 18 bp poly T stretch, typically conserved in Lepidoptera. PMID:24893878

  4. Proteomic Analysis of Silkworm Antennae.

    PubMed

    Zhao, Yunpo; Li, Haichao; Miao, Xuexia

    2015-11-01

    The silkworm Bombyx mori is an oligophagous insect that feeds mainly on mulberry leaves. The olfactory system of silkworm is a good model to study olfaction in Lepidoptera. Here, we carried out shotgun proteomic analysis and MS sequencing of the silkmoth antennae. A total of 364 proteins were detected, 77 were female specific, 143 were male specific, and 144 were expressed in both male and female antennae. Five odorant-binding proteins, two chemosensory proteins, and one olfactory receptor were identified. They may play a major role in the perception of odorants. An esterase and an aldehyde dehydrogenase were found only in male antennae. Glutathione S-transferases (GSTs) and cytochrome P450s, also found in silkworm antennae, may be involved in the degradation of xenobiotics. Additionally, antioxidation proteins and immunity proteins were identified. Juvenile hormone binding proteins (JHBP), juvenile hormone resistance protein II, and juvenile hormone episode hydrolase (JHEH) were found in the proteomic analysis, which suggests that the antennae are a target for juvenile hormone in the silkworm. Our results provide insight into the expression of proteins in the antennae of silkworm and will facilitate the future functional analysis of silkworm antennae. PMID:26515890

  5. Inhibition of Carrageenan-Induced Acute Inflammation in Mice by Oral Administration of Anthocyanin Mixture from Wild Mulberry and Cyanidin-3-Glucoside

    PubMed Central

    Hassimotto, Neuza Mariko Aymoto; Moreira, Vanessa; do Nascimento, Neide Galvão; Souto, Pollyana Cristina Maggio de Castro; Teixeira, Catarina; Lajolo, Franco Maria

    2013-01-01

    Anthocyanins are flavonoids which demonstrated biological activities in in vivo and in vitro models. Here in the anti-inflammatory properties of an anthocyanin-enriched fraction (AF) extracted from wild mulberry and the cyanidin-3-glucoside (C3G), the most abundant anthocyanin in diet, were studied in two acute inflammation experimental models, in the peritonitis and in the paw oedema assays, both of which were induced by carrageenan (cg) in mice. In each trial, AF and C3G (4 mg/100 g/animal) were orally administered in two distinct protocols: 30 min before and 1 h after cg stimulus. The administration of both AF and C3G suppresses the paw oedema in both administration times (P < 0.05). In the peritonitis, AF and C3G reduced the polymorphonuclear leukocytes (PMN) influx in the peritoneal exudates when administered 1 h after cg injection. AF was more efficient reducing the PMN when administered 30 min before cg. Both AF and C3G were found to suppress mRNA as well as protein levels of COX-2 upregulated by cg in both protocols, but the inhibitory effect on PGE2 production in the peritoneal exudates was observed when administered 30 min before cg (P < 0.05). Our findings suggest that AF and C3G minimize acute inflammation and they present positive contributions as dietary supplements. PMID:23484081

  6. The expression analysis of cysteine proteinase-like protein in wild-type and nm2 mutant silkworm (Lepidoptera: Bombyx mori).

    PubMed

    Wu, Fan; Kang, Lequn; Wang, Pingyang; Zhao, Qiaoling

    2016-07-15

    The mutant of non-molting in the 2nd instar (nm2) is a recently discovered mutant of Bombyx mori. The mutant cannot molt and exuviate and died successively in premolting of 2nd instar. In this study, two dimensional gel electrophoresis (2-DE) was performed to screen the differential expression of epidermis proteins in pre-molting larvae of 2nd instar between the wild-type and nm2 mutant. Interestingly, a cysteine proteinase-like (BmCP-like) protein in nm2 was significantly higher than that of the wild-type. The transcription profiles of BmCP-like gene were investigated by quantitative real-time PCR (qRT-PCR), and the result revealed that BmCP-like mRNA was remarkably higher in nm2 than that of the wild-type. The transcription level of BmCP-like was high in the epidermis while low in the midgut and hemocytes, and fluctuate with development, while the highest in the newly molted larvae of 3rd and lowest in the pre-molting of the 1st and 2nd instar. The body of injected BmCP-like RNAi of 2nd larvae formed a dark spots around the injection place. These results suggested the BmCP-like gene play a key role in the degradation of the cuticle and epidermis layer during molting of 1st and 2nd instar silkworm. Furthermore, the ORF of BmCP-like gene in nm2 was the same to the wild-type. These studies give us a hint that BmCP-like gene maybe not the major gene responsible for nm2, but BmCP-like gene might participate in the immune systems of silkworm, and the upregulation of BmCP-like transcription in the nm2 mutant might be induced by the disadvantages that limit the growth and development of silkworm in order to survive. PMID:27080953

  7. A feasibility study towards cryopreservation of silkworm eggs: response of non-diapause silkworm eggs to low temperature.

    PubMed

    Jingade, Anuradha H; Lekha, G; Srinivasa, Babu G K; Manjula, A; Sivaprasad, V

    2015-01-01

    Insect embryos are very sensitive to chilling temperatures and vary with species and their developmental stages. Insect eggs are small and can supercool to temperatures ranging from -5 degrees C to -40 degrees C. In general, insects rely on a variety of ecological and physiological adaptations to survive low temperatures, making cryopreservation technique significantly complex. Mulberry silkworm (Bombyx mori L., Lepidoptera: Bombycidae) eggs are cleidoic with chorion (approximately 20-25 μm thick). Preservation of non-diapause eggs to a limited period is practiced usually to delay hatching. The advantage of early embryonic periods having resistance to low temperature is utilized for chilling of eggs and preservation for long periods. However, technique for cryopreservation of silkworm eggs is not yet developed and the identification of precise embryonic stage and chill sensitivity is necessary for effective silkworm cryopreservation. The paper reports the chill-sensitivity and tolerance of non-diapause silkworm embryos of mulberry silkworm at various embryonic stages. Silkworm embryo of 48h-age are relatively chill-sensitive as compared to other embryonic ages. This is vital information for the development of cryopreservation protocol for silkworm eggs. PMID:26017168

  8. Free-radical scavenging properties of low molecular weight peptide(s) isolated from S1 cultivar of mulberry leaves and their impact on Bombyx mori (L.) (Bombycidae)

    PubMed Central

    2014-01-01

    The mulberry leaves have been considered as a sole food source for silkworm, Bombyx mori (L.). In present work an attempt was made to investigate the role of low molecular weight peptide(s) isolated from mulberry leaves on silkworm rearing. Also we have tried to find out the role of free-radical scavenging activities of isolated peptide(s) on silkworm growth. Larval growth rate was found effective under the influence of peptide(s). Consumption rate of larvae after peptide(s) treatment on mulberry leaves was significantly enhanced over control. High antioxidant activity was found in Low molecular weight peptide(s) which have an effect on silkworm. PMID:24612781

  9. Identification of 2chromosome region translocated onto the W chromosome by RFLP with EST-cDNA clones in the Gensei-kouken strains of the mulberry silkworm, Bombyx mori L.

    PubMed

    Sreekumar, Sivaramakurup; Kadono-Okuda, Keiko; Nagayasu, Ken-Ichi; Hara, Wajiro

    2010-01-01

    In silkworms, sex-limited strains are either obtained spontaneously or induced by X-rays or gamma rays. When a fragment of an autosome carrying a dominant allele of those genes responsible for certain characters is translocated onto a W chromosome, the female of the successive generations will express these phenotypic characters and sex discrimination can be facilitated. Gensei-kouken strains are sex-limited strains of silkworms developed by irradiating the pupae with gamma rays, by which a portion of the second chromosome is translocated onto the W chromosome. In these improved strains, the females are yellow-blooded and spin yellow cocoons. By using the EST-cDNA clones mapped on the Z chromosome, we identified the sex according to the polymorphic banding pattern or intensity of the signals. Furthermore, by using the clones on the second chromosome, the region of the second chromosome translocated onto the W chromosome was also defined. In both the A95 and A 96 strains selected for the present study, only the mid-portion of the second chromosome was translocated. The differences in length of the fragments translocated in these strains are discussed. PMID:21637601

  10. The Origin and Dispersal of the Domesticated Chinese Oak Silkworm, Antheraea pernyi, in China: A Reconstruction Based on Ancient Texts

    PubMed Central

    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

  11. Mapping and recombination analysis of two moth colour mutations, Black moth and Wild wing spot, in the silkworm Bombyx mori.

    PubMed

    Ito, K; Katsuma, S; Kuwazaki, S; Jouraku, A; Fujimoto, T; Sahara, K; Yasukochi, Y; Yamamoto, K; Tabunoki, H; Yokoyama, T; Kadono-Okuda, K; Shimada, T

    2016-01-01

    Many lepidopteran insects exhibit body colour variations, where the high phenotypic diversity observed in the wings and bodies of adults provides opportunities for studying adaptive morphological evolution. In the silkworm Bombyx mori, two genes responsible for moth colour mutation, Bm and Ws, have been mapped to 0.0 and 14.7 cM of the B. mori genetic linkage group 17; however, these genes have not been identified at the molecular level. We performed positional cloning of both genes to elucidate the molecular mechanisms that underlie the moth wing- and body-colour patterns in B. mori. We successfully narrowed down Bm and Ws to ~2-Mb-long and 100-kb-long regions on the same scaffold Bm_scaf33. Gene prediction analysis of this region identified 77 candidate genes in the Bm region, whereas there were no candidate genes in the Ws region. Fluorescence in-situ hybridisation analysis in Bm mutant detected chromosome inversion, which explains why there are no recombination in the corresponding region. The comparative genomic analysis demonstrated that the candidate regions of both genes shared synteny with a region associated with wing- and body-colour variations in other lepidopteran species including Biston betularia and Heliconius butterflies. These results suggest that the genes responsible for wing and body colour in B. mori may be associated with similar genes in other Lepidoptera. PMID:26219230

  12. MnTEdb, a collective resource for mulberry transposable elements

    PubMed Central

    Ma, Bi; Li, Tian; Xiang, Zhonghuai; He, Ningjia

    2015-01-01

    Mulberry has been used as an economically important food crop for the domesticated silkworm for thousands of years, resulting in one of the oldest and well-known plant-herbivore interactions. The genome of Morus notabilis has now been sequenced and there is an opportunity to mine the transposable element (TE) data. To better understand the roles of TEs in structural, functional and evolutionary dynamics of the mulberry genome, a specific, comprehensive and user-friendly web-based database, MnTEdb, was constructed. It was built based on a detailed and accurate identification of all TEs in mulberry. A total of 5925 TEs belonging to 13 superfamilies and 1062 families were deposited in this database. MnTEdb enables users to search, browse and download the mulberry TE sequences. Meanwhile, data mining tools, including BLAST, GetORF, HMMER, Sequence Extractor and JBrowse were also integrated into MnTEdb. MnTEdb will assist researchers to efficiently take advantage of our newly annotated TEs, which facilitate their studies in the origin, amplification and evolution of TEs, as well as the comparative analysis among the different species. Database URL: http://morus.swu.edu.cn/mntedb/ PMID:25725060

  13. MnTEdb, a collective resource for mulberry transposable elements.

    PubMed

    Ma, Bi; Li, Tian; Xiang, Zhonghuai; He, Ningjia

    2015-01-01

    Mulberry has been used as an economically important food crop for the domesticated silkworm for thousands of years, resulting in one of the oldest and well-known plant-herbivore interactions. The genome of Morus notabilis has now been sequenced and there is an opportunity to mine the transposable element (TE) data. To better understand the roles of TEs in structural, functional and evolutionary dynamics of the mulberry genome, a specific, comprehensive and user-friendly web-based database, MnTEdb, was constructed. It was built based on a detailed and accurate identification of all TEs in mulberry. A total of 5925 TEs belonging to 13 superfamilies and 1062 families were deposited in this database. MnTEdb enables users to search, browse and download the mulberry TE sequences. Meanwhile, data mining tools, including BLAST, GetORF, HMMER, Sequence Extractor and JBrowse were also integrated into MnTEdb. MnTEdb will assist researchers to efficiently take advantage of our newly annotated TEs, which facilitate their studies in the origin, amplification and evolution of TEs, as well as the comparative analysis among the different species. Database URL: http://morus.swu.edu.cn/mntedb/ PMID:25725060

  14. 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

  15. 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

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

    PubMed

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

    2015-06-01

    Previously, it was reported that Enterococcus mundtii (E. mundtii) was associated with flacherie disease of silkworm larvae reared on artificial diet. In this study, we report that E. mundtii was isolated from diseased silkworm larvae, and validated as a pathogenic bacterium of the animal. When silkworm larva was infected with 1.04 × 10⁶ colony-forming units of E. mundtii via oral administration of diet, half population died within six days, indicating that the bacterium is pathogenic to silkworm. Less severe infection was found to cause anorexia and hamper the development of larvae. This pathogen was found to proliferate in both time- and dose-dependent manner in the gastrointestinal tract of the animal. The bacterium was isolated from powder of artificial diet made from mulberry leaves, and from mulberry leaves growing at a field. Minimum inhibitory concentration determination revealed that this bacterium was susceptible to tested antibiotics. Vancomycin treatment of diet significantly decreased the number of E. mundtii in intestine of silkworm larvae infected with the bacteria, compared to control. Furthermore, autoclaving or gamma ray irradiation of diet was also effective for exclusion of E. mundtii from the diet without the loss of its nutrient capacities. These results suggest that mulberry leaves used in making artificial diet for silkworm larvae is one of the sources of E. mundtii infection; and that antibiotic treatment, autoclaving or gamma ray irradiation of artificial diet can exclude the bacteria. PMID:26193940

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

    NASA Astrophysics Data System (ADS)

    Liang, Xue; Liu, lh64. Hong

    2012-07-01

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

  18. Molecular characterization and expression analysis of the mulberry Na(+)/H(+) exchanger gene family.

    PubMed

    Cao, Boning; Long, Dingpei; Zhang, Meng; Liu, Changying; Xiang, Zhonghuai; Zhao, Aichun

    2016-02-01

    Na(+)/H(+) exchangers (NHXs) have important roles in cellular pH, and Na(+) and K(+) homeostasis in plants. Mulberry is not only an important traditional economic woody plant known for its leaves, which are the exclusive food source of the silkworm Bombyx mori, but it can also adapt to many different adverse conditions, including saline environments. However, little is known about the NHXs in this important perennial tree. In this study, we identified and cloned seven putative NHX gene family members from Morus atropurpurea based on a genome-wide analysis of the Morus genome database. A phylogenetic analysis and genomic organization of mulberry NHXs suggested that the mulberry NHX family forms three distinct subgroups. Transcriptome data and real-time PCR of different mulberry varieties under normal culture conditions revealed that the mulberry NHX family has a different tissue-specific pattern in the two mulberry species. The MaNHX genes' expression analyses under different stresses (salt and drought) and signal molecules (abscisic acid, salicylic acid, hydrogen peroxide and methyl jasmonate) revealed that MaNHXs not only could be induced by salt, drought and abscisic acid as describe in the literature, but were also induced by other signal molecules, which indicated that MaNHX members exhibited diverse and complicated expression patterns in different mulberry tissues under various abiotic stresses, phytohormones and plant signaling molecules. Our results provide some insights into new and emerging cellular and physiological functions of this group of H(+)-coupled cation exchangers, beyond their function in salt tolerance, and also provide the basis for further characterizations of MaNHXs' physiological functions. PMID:26730882

  19. Identification of the mulberry genes involved in ethylene biosynthesis and signaling pathways and the expression of MaERF-B2-1 and MaERF-B2-2 in the response to flooding stress.

    PubMed

    Shang, Jingzhe; Song, Penghua; Ma, Bi; Qi, Xiwu; Zeng, Qiwei; Xiang, Zhonghuai; He, Ningjia

    2014-12-01

    The phytohormone ethylene is essential to plant growth and development. It plays crucial roles in responses to biotic and abiotic stress. The mulberry tree is an important crop plant in countries in which people rear silkworms for silk production. The availability of the mulberry genome has made it possible to identify mulberry genes involved in ethylene biosynthesis and signal pathways. A total of 145 mulberry genes were identified by both homology-based and hidden Markov model (HMM) search, including 29 genes associated with ethylene biosynthesis and 116 genes in the AP2/ERF family. Studies on gene structure have provided a genetic basis for understanding the functions of these genes. The differences in gene expression were also observed in different tissues. The expression of two mulberry genes in the AP2/ERF family, MaERF-B2-1 and MaERF-B2-2, was found to be associated with the response to flooding stress. PMID:25231943

  20. 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.

  1. SilkDB: a knowledgebase for silkworm biology and genomics.

    PubMed

    Wang, Jing; Xia, Qingyou; He, Ximiao; Dai, Mingtao; Ruan, Jue; Chen, Jie; Yu, Guo; Yuan, Haifeng; Hu, Yafeng; Li, Ruiqiang; Feng, Tao; Ye, Chen; Lu, Cheng; Wang, Jun; Li, Songgang; Wong, Gane Ka-Shu; Yang, Huanming; Wang, Jian; Xiang, Zhonghuai; Zhou, Zeyang; Yu, Jun

    2005-01-01

    The Silkworm Knowledgebase (SilkDB) is a web-based repository for the curation, integration and study of silkworm genetic and genomic data. With the recent accomplishment of a approximately 6X draft genome sequence of the domestic silkworm (Bombyx mori), SilkDB provides an integrated representation of the large-scale, genome-wide sequence assembly, cDNAs, clusters of expressed sequence tags (ESTs), transposable elements (TEs), mutants, single nucleotide polymorphisms (SNPs) and functional annotations of genes with assignments to InterPro domains and Gene Ontology (GO) terms. SilkDB also hosts a set of ESTs from Bombyx mandarina, a wild progenitor of B.mori, and a collection of genes from other Lepidoptera. Comparative analysis results between the domestic and wild silkworm, between B.mori and other Lepidoptera, and between B.mori and the two sequenced insects, fruitfly and mosquito, are displayed by using B.mori genome sequence as a reference framework. Designed as a basic platform, SilkDB strives to provide a comprehensive knowledgebase about the silkworm and present the silkworm genome and related information in systematic and graphical ways for the convenience of in-depth comparative studies. SilkDB is publicly accessible at http://silkworm.genomics.org.cn. PMID:15608225

  2. Mutation of a cuticular protein, BmorCPR2, alters larval body shape and adaptability in silkworm, Bombyx mori.

    PubMed

    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

    2014-04-01

    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

  3. 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

  4. Identification and analysis of the pigment composition and sources in the colored cocoon of the silkworm, Bombyx mori, by HPLC-DAD.

    PubMed

    Zhu, Lin; Zhang, Yu-Qing

    2014-01-01

    This study used the larval tissues and colored cocoons of silkworms, Bombyx mori L. (Lepidoptera: Bombycidae), that were fed leaves of cultivated mulberry, Husang 32, as experimental material. The pigment composition and content in colored cocoons and tissues of the 5th instar larvae and the mulberry leaves were rapidly detected using organic solvent extraction and reverse phase high-performance liquid chromatography with diode array detection. It was found that the mulberry leaf mainly contained four types of pigment: lutein (30.86%), β-carotene (26.3%), chlorophyll a (24.62%), and chlorophyll b (18.21%). The silk glands, blood, and cocoon shells of six yellow-red cocoons were used as the experimental materials. The results showed that there were generally two kinds of carotenoids (lutein and β-carotene) in the silk gland and cocoon shell, a little violaxanthin was detected in silk gland, and the pigment found in the blood was mainly lutein in all varieties of silkworm tested. Chlorophyll a and b had not been digested and utilized in the yellow-red series of silkworm. The method used to detect visible pigments reported here could be used to breed new colors of cocoons and to develop and utilize the pigments found in mulberry. PMID:25373178

  5. Identification and Analysis of the Pigment Composition and Sources in the Colored Cocoon of the Silkworm, Bombyx mori, by HPLC-DAD

    PubMed Central

    Zhu, Lin; Zhang, Yu-Qing

    2014-01-01

    This study used the larval tissues and colored cocoons of silkworms, Bombyx mori L. (Lepidoptera: Bombycidae), that were fed leaves of cultivated mulberry, Husang 32, as experimental material. The pigment composition and content in colored cocoons and tissues of the 5th instar larvae and the mulberry leaves were rapidly detected using organic solvent extraction and reverse phase high-performance liquid chromatography with diode array detection. It was found that the mulberry leaf mainly contained four types of pigment: lutein (30.86%), β-carotene (26.3%), chlorophyll a (24.62%), and chlorophyll b (18.21%). The silk glands, blood, and cocoon shells of six yellow-red cocoons were used as the experimental materials. The results showed that there were generally two kinds of carotenoids (lutein and β-carotene) in the silk gland and cocoon shell, a little violaxanthin was detected in silk gland, and the pigment found in the blood was mainly lutein in all varieties of silkworm tested. Chlorophyll a and b had not been digested and utilized in the yellow-red series of silkworm. The method used to detect visible pigments reported here could be used to breed new colors of cocoons and to develop and utilize the pigments found in mulberry. PMID:25373178

  6. Genome-wide patterns of genetic variation among silkworms.

    PubMed

    Zhang, Xingtan; Nie, Mengyun; Zhao, Qian; Wu, Yuqian; Wang, Genhong; Xia, Qingyou

    2015-08-01

    Although the draft genome sequence of silkworm is available for a decade, its genetic variations, especially structural variations, are far from well explored. In this study, we identified 1,298,659 SNPs and 9,731 indels, of which 32 % of SNPs and 92.2 % of indels were novel compared to previous silkworm re-sequencing analysis. In addition, we applied a read depth-based approach to investigate copy number variations among 21 silkworm strains at genome-wide level. This effort resulted in 562 duplicated and 41 deleted CNV regions, and among them 442 CNV were newly identified. Functional annotation of genes affected by these genetic variations reveal that these genes include a wide spectrum of molecular functions, such as immunity and drug detoxification, which are important for the adaptive evolution of silkworms. We further validated the predicted CNV regions using q-PCR. 94.7 % (36/38) of the selected regions show divergent copy numbers compared to a single-copy gene OR2. In addition, potential presence/absence variations are also observed in our study: 11 genes are present in the reference genome, but absent in other strains. Overall, we draw an integrative map of silkworm genetic variation at genome-wide level. The identification of genetic variations in this study improves our understanding that these variants play important roles in shaping phenotypic variations between wild and domesticated silkworms. PMID:25749967

  7. Demographic history and gene flow during silkworm domestication

    PubMed Central

    2014-01-01

    Background Gene flow plays an important role in domestication history of domesticated species. However, little is known about the demographic history of domesticated silkworm involving gene flow with its wild relative. Results In this study, four model-based evolutionary scenarios to describe the demographic history of B. mori were hypothesized. Using Approximate Bayesian Computation method and DNA sequence data from 29 nuclear loci, we found that the gene flow at bottleneck model is the most likely scenario for silkworm domestication. The starting time of silkworm domestication was estimated to be approximate 7,500 years ago; the time of domestication termination was 3,984 years ago. Using coalescent simulation analysis, we also found that bi-directional gene flow occurred during silkworm domestication. Conclusions Estimates of silkworm domestication time are nearly consistent with the archeological evidence and our previous results. Importantly, we found that the bi-directional gene flow might occur during silkworm domestication. Our findings add a dimension to highlight the important role of gene flow in domestication of crops and animals. PMID:25123546

  8. 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.

  9. Accumulation of Rutin and Betulinic Acid and Expression of Phenylpropanoid and Triterpenoid Biosynthetic Genes in Mulberry (Morus alba L.).

    PubMed

    Zhao, Shicheng; Park, Chang Ha; Li, Xiaohua; Kim, Yeon Bok; Yang, Jingli; Sung, Gyoo Byung; Park, Nam Il; Kim, Soonok; Park, Sang Un

    2015-09-30

    Mulberry (Morus alba L.) is used in traditional Chinese medicine and is the sole food source of the silkworm. Here, 21 cDNAs encoding phenylpropanoid biosynthetic genes and 21 cDNAs encoding triterpene biosynthetic genes were isolated from mulberry. The expression levels of genes involved in these biosynthetic pathways and the accumulation of rutin, betulin, and betulinic acid, important secondary metabolites, were investigated in different plant organs. Most phenylpropanoid and triterpene biosynthetic genes were highly expressed in leaves and/or fruit, and most genes were downregulated during fruit ripening. The accumulation of rutin was more than fivefold higher in leaves than in other organs, and higher levels of betulin and betulinic acid were found in roots and leaves than in fruit. By comparing the contents of these compounds with gene expression levels, we speculate that MaUGT78D1 and MaLUS play important regulatory roles in the rutin and betulin biosynthetic pathways. PMID:26343778

  10. 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

  11. Quantification and Purification of Mulberry Anthocyanins With Macroporous Resins

    PubMed Central

    Liu, Xueming

    2004-01-01

    Total anthocyanins in different cultivars of mulberry were measured and a process for the industrial preparation of mulberry anthocyanins as a natural food colorant was studied. In 31 cultivars of mulberry, the total anthocyanins, calculated as cyanidin 3-glucoside, ranged from 147.68 to 2725.46?mg/L juice. Extracting and purifying with macroporous resins was found to be an efficient potential method for the industrial production of mulberry anthocyanins as a food colorant. Of six resins tested, X-5 demonstrated the best adsorbent capability for mulberry anthocyanins (91?mg/mL resin). The adsorption capacity of resins increased with the surface area and the pore radius. Residual mulberry fruit juice after extraction of pigment retained most of its nutrients, except for anthocyanins, and may provide a substrate for further processing. PMID:15577197

  12. Quantification and Purification of Mulberry Anthocyanins with Macroporous Resins.

    PubMed

    Liu, Xueming; Xiao, Gengsheng; Chen, Weidong; Xu, Yujuan; Wu, Jijun

    2004-01-01

    Total anthocyanins in different cultivars of mulberry were measured and a process for the industrial preparation of mulberry anthocyanins as a natural food colorant was studied. In 31 cultivars of mulberry, the total anthocyanins, calculated as cyanidin 3-glucoside, ranged from $147.68$ to $2725.46$ mg/L juice. Extracting and purifying with macroporous resins was found to be an efficient potential method for the industrial production of mulberry anthocyanins as a food colorant. Of six resins tested, X-5 demonstrated the best adsorbent capability for mulberry anthocyanins (91 mg/mL resin). The adsorption capacity of resins increased with the surface area and the pore radius. Residual mulberry fruit juice after extraction of pigment retained most of its nutrients, except for anthocyanins, and may provide a substrate for further processing. PMID:15577197

  13. The genome of a lepidopteran model insect, the silkworm Bombyx mori.

    PubMed

    2008-12-01

    Bombyx mori, the domesticated silkworm, is a major insect model for research, and the first lepidopteran for which draft genome sequences became available in 2004. Two independent data sets from whole-genome shotgun sequencing were merged and assembled together with newly obtained fosmid- and BAC-end sequences. The remarkably improved new assembly is presented here. The 8.5-fold sequence coverage of an estimated 432 Mb genome was assembled into scaffolds with an N50 size of approximately 3.7 Mb; the largest scaffold was 14.5 million base pairs. With help of a high-density SNP linkage map, we anchored 87% of the scaffold sequences to all 28 chromosomes. A particular feature was the high repetitive sequence content estimated to be 43.6% and that consisted mainly of transposable elements. We predicted 14,623 gene models based on a GLEAN-based algorithm, a more accurate prediction than the previous gene models for this species. Over three thousand silkworm genes have no homologs in other insect or vertebrate genomes. Some insights into gene evolution and into characteristic biological processes are presented here and in other papers in this issue. The massive silk production correlates with the existence of specific tRNA clusters, and of several sericin genes assembled in a cluster. The silkworm's adaptation to feeding on mulberry leaves, which contain toxic alkaloids, is likely linked to the presence of new-type sucrase genes, apparently acquired from bacteria. The silkworm genome also revealed the cascade of genes involved in the juvenile hormone biosynthesis pathway, and a large number of cuticular protein genes. PMID:19121390

  14. Analysis of midgut gene expression profiles from different silkworm varieties after exposure to high temperature.

    PubMed

    Li, Qing Rong; Xiao, Yang; Wu, Fu Quan; Ye, Ming Qiang; Luo, Guo Qing; Xing, Dong Xu; Li, Li; Yang, Qiong

    2014-10-01

    The silkworm is a poikilothermic animal, whose growth and development is significantly influenced by environmental temperature. To identify genes and metabolic pathways involved in the heat-stress response, digital gene expression analysis was performed on the midgut of the thermotolerant silkworm variety '932' and thermosensitive variety 'HY' after exposure to high temperature (932T and HYT). Deep sequencing yielded 6,211,484, 5,898,028, 5,870,395 and 6,088,303 reads for the 932, 932T, HY and HYT samples, respectively. The annotated genes associated with these tags numbered 4357, 4378, 4296 and 4658 for the 932, 932T, HY and HYT samples, respectively. In the HY-vs-932, 932-vs-932T, and HY-vs-HYT comparisons, 561, 316 and 281 differentially expressed genes were identified, which could be assigned to 179, 140 and 123 biological pathways, respectively. It was found that some of the biological pathways, which included oxidative phosphorylation, related to glucose and lipid metabolism, are greatly affected by high temperature and may lead to a decrease in the ingestion of fresh mulberry. When subjected to an early period of continuous heat stress, HSP genes, such as HSP19.9, HSP23.7, HSP40-3, HSP70, HSP90 and HSP70 binding protein, are up-regulated but then reduced after 24h and the thermotolerant '932' strain has higher levels of mRNA of some HSPs, except HSP70, than the thermosensitive variety during continuous high temperature treatment. It is suggested that HSPs and the levels of their expression may play important roles in the resistance to high temperature stress among silkworm varieties. This study has generated important reference tools that can be used to further analyze the mechanisms that underlie thermotolerance differences among silkworm varieties. PMID:25046138

  15. 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

  16. Consumption of Bt rice pollen containing Cry1C or Cry2A protein poses a low to negligible risk to the silkworm Bombyx mori (Lepidoptera: Bombyxidae).

    PubMed

    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/cm(2) 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/cm(2) 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

  17. An Approach to Function Annotation for Proteins of Unknown Function (PUFs) in the Transcriptome of Indian Mulberry.

    PubMed

    Dhanyalakshmi, K H; Naika, Mahantesha B N; Sajeevan, R S; Mathew, Oommen K; Shafi, K Mohamed; Sowdhamini, Ramanathan; N Nataraja, Karaba

    2016-01-01

    The modern sequencing technologies are generating large volumes of information at the transcriptome and genome level. Translation of this information into a biological meaning is far behind the race due to which a significant portion of proteins discovered remain as proteins of unknown function (PUFs). Attempts to uncover the functional significance of PUFs are limited due to lack of easy and high throughput functional annotation tools. Here, we report an approach to assign putative functions to PUFs, identified in the transcriptome of mulberry, a perennial tree commonly cultivated as host of silkworm. We utilized the mulberry PUFs generated from leaf tissues exposed to drought stress at whole plant level. A sequence and structure based computational analysis predicted the probable function of the PUFs. For rapid and easy annotation of PUFs, we developed an automated pipeline by integrating diverse bioinformatics tools, designated as PUFs Annotation Server (PUFAS), which also provides a web service API (Application Programming Interface) for a large-scale analysis up to a genome. The expression analysis of three selected PUFs annotated by the pipeline revealed abiotic stress responsiveness of the genes, and hence their potential role in stress acclimation pathways. The automated pipeline developed here could be extended to assign functions to PUFs from any organism in general. PUFAS web server is available at http://caps.ncbs.res.in/pufas/ and the web service is accessible at http://capservices.ncbs.res.in/help/pufas. PMID:26982336

  18. An Approach to Function Annotation for Proteins of Unknown Function (PUFs) in the Transcriptome of Indian Mulberry

    PubMed Central

    Dhanyalakshmi, K. H.; Naika, Mahantesha B. N.; Sajeevan, R. S.; Mathew, Oommen K.; Shafi, K. Mohamed; Sowdhamini, Ramanathan; N. Nataraja, Karaba

    2016-01-01

    The modern sequencing technologies are generating large volumes of information at the transcriptome and genome level. Translation of this information into a biological meaning is far behind the race due to which a significant portion of proteins discovered remain as proteins of unknown function (PUFs). Attempts to uncover the functional significance of PUFs are limited due to lack of easy and high throughput functional annotation tools. Here, we report an approach to assign putative functions to PUFs, identified in the transcriptome of mulberry, a perennial tree commonly cultivated as host of silkworm. We utilized the mulberry PUFs generated from leaf tissues exposed to drought stress at whole plant level. A sequence and structure based computational analysis predicted the probable function of the PUFs. For rapid and easy annotation of PUFs, we developed an automated pipeline by integrating diverse bioinformatics tools, designated as PUFs Annotation Server (PUFAS), which also provides a web service API (Application Programming Interface) for a large-scale analysis up to a genome. The expression analysis of three selected PUFs annotated by the pipeline revealed abiotic stress responsiveness of the genes, and hence their potential role in stress acclimation pathways. The automated pipeline developed here could be extended to assign functions to PUFs from any organism in general. PUFAS web server is available at http://caps.ncbs.res.in/pufas/ and the web service is accessible at http://capservices.ncbs.res.in/help/pufas. PMID:26982336

  19. 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

  20. [Effects of sub-lethal dosages abamectin on food intake and digestive enzyme activities of silkworm Bombyx mori L].

    PubMed

    Zhu, Jiu-sheng; Wang, Jing; Gao, Hai-yan; Qin, Shu; Qiao, Xiong-wu; Han, Ju-cai

    2008-11-01

    Mulberry leaves treated with sub-lethal dosages (LC5, LC10 and LC20) abameetin were fed to the 5th instar larvae of silkworm (Bombyx mori L.), and the food intake and digestive enzyme activities of the larvae were studied by using gravimetric method and measuring enzyme activities. The results showed that sub-lethal dosages abameetin significantly inhibited the growth and food intake of the larvae, with their body mass and its increase rate as well as their relative growth rate being significantly lower than the control, and accompanied with the decreases of food intake, its relative consumption rate, and feces amount. The efficiency of the conversion of ingested food (ECI) and that of the conversion of digested food (EDI) also reduced, but the approximate digestibility (AD) increased significantly. The amylase and sucrase activities in the midgut of the larvae treated with abameetin decreased significantly for a longer time at the beginning, and then recovered to the same as or a higher level than the control, whereas the trehalase activity decreased significantly for a shorter time at the beginning, then increased significantly, and finally recovered to the normal. It was suggested that sub-lethal dosages abameetin had definite toxicity to the silkworm, and the toxic effect was increased with increasing dosage, which could result in the turbulence of silkworm's digestive system, and further, affect its food intake and its growth and development. PMID:19238858

  1. 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...

  2. Spectral imaging analysis for silkworm gender classification

    NASA Astrophysics Data System (ADS)

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

    2013-05-01

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

  3. 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%.

  4. Complete mitochondrial genome of the mulberry white caterpillar Rondotia menciana (Lepidoptera: Bombycidae).

    PubMed

    Kim, Min Jee; Jun, Jumin; Kim, Iksoo

    2016-01-01

    The mulberry white caterpillar, Rondotia menciana, belongs to the lepidopteran family Bombycidae, in which the domestic silkworm, Bombyx mori is included. In this study, we describe the complete mitochondrial genome of R. menciana in terms of general genomic features and characteristic features found in the A+T-rich region. The 15,364 bp long genome consisted of a typical set of genes (13 protein-coding genes [PCGs], 2 rRNA genes, and 22 tRNA genes) and 1 major non-coding A+T-rich region, with the typical arrangement found in Lepidoptera. Twelve of the 13 PCGs started with typical ATN codons, except for the COI, which began with CGA and twelve of 13 PCGs had complete stop codons, except for the COII, which ended with a single T. The 360 bp long A+T-rich region harbored the conserved sequence blocks typically found in lepidopteran insects. Additionally, the A+T-rich region of R. menciana contained one tRNA(Met)-like structure, which had a proper anticodon and secondary structure. PMID:24779598

  5. Effects of Altered Catecholamine Metabolism on Pigmentation and Physical Properties of Sclerotized Regions in the Silkworm Melanism Mutant

    PubMed Central

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

    2012-01-01

    Catecholamine metabolism plays an important role in the determination of insect body color and cuticle sclerotization. To date, limited research has focused on these processes in silkworm. In the current study, we analyzed the interactions between catecholamines and melanin genes and their effects on the pigmentation patterns and physical properties of sclerotized regions in silkworm, using the melanic mutant melanism (mln) silkworm strain as a model. Injection of ?-alanine into mln mutant silkworm induced a change in catecholamine metabolism and turned its body color yellow. Further investigation of the catecholamine content and expression levels of the corresponding melanin genes from different developmental stages of Dazao-mln (mutant) and Dazao (wild-type) silkworm revealed that at the larval and adult stages, the expression patterns of melanin genes precipitated dopamine accumulation corresponding to functional loss of Bm-iAANAT, a repressive effect of excess NBAD on ebony, and upregulation of tan in the Dazao-mln strain. During the early pupal stage, dopamine did not accumulate in Dazao-mln, since upregulation of ebony and black genes led to conversion of high amounts of dopamine into NBAD, resulting in deep yellow cuticles. Scanning electron microscope analysis of a cross-section of adult dorsal plates from both wild-type and mutant silkworm disclosed the formation of different layers in Dazao-mln owing to lack of NADA, compared to even and dense layers in Dazao. Analysis of the mechanical properties of the anterior wings revealed higher storage modulus and lower loss tangent in Dazao-mln, which was closely associated with the altered catecholamine metabolism in the mutant strain. Based on these findings, we conclude that catecholamine metabolism is crucial for the color pattern and physical properties of cuticles in silkworm. Our results should provide a significant contribution to Lepidoptera cuticle tanning research. PMID:22937004

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

    PubMed

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

    2012-01-01

    Catecholamine metabolism plays an important role in the determination of insect body color and cuticle sclerotization. To date, limited research has focused on these processes in silkworm. In the current study, we analyzed the interactions between catecholamines and melanin genes and their effects on the pigmentation patterns and physical properties of sclerotized regions in silkworm, using the melanic mutant melanism (mln) silkworm strain as a model. Injection of β-alanine into mln mutant silkworm induced a change in catecholamine metabolism and turned its body color yellow. Further investigation of the catecholamine content and expression levels of the corresponding melanin genes from different developmental stages of Dazao-mln (mutant) and Dazao (wild-type) silkworm revealed that at the larval and adult stages, the expression patterns of melanin genes precipitated dopamine accumulation corresponding to functional loss of Bm-iAANAT, a repressive effect of excess NBAD on ebony, and upregulation of tan in the Dazao-mln strain. During the early pupal stage, dopamine did not accumulate in Dazao-mln, since upregulation of ebony and black genes led to conversion of high amounts of dopamine into NBAD, resulting in deep yellow cuticles. Scanning electron microscope analysis of a cross-section of adult dorsal plates from both wild-type and mutant silkworm disclosed the formation of different layers in Dazao-mln owing to lack of NADA, compared to even and dense layers in Dazao. Analysis of the mechanical properties of the anterior wings revealed higher storage modulus and lower loss tangent in Dazao-mln, which was closely associated with the altered catecholamine metabolism in the mutant strain. Based on these findings, we conclude that catecholamine metabolism is crucial for the color pattern and physical properties of cuticles in silkworm. Our results should provide a significant contribution to Lepidoptera cuticle tanning research. PMID:22937004

  7. 20. GROVE OF TREES PINES, MULBERRY, JUNIPER, BLUE SPRUCE ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    20. GROVE OF TREES -- PINES, MULBERRY, JUNIPER, BLUE SPRUCE -- TRANSPLANTED FROM NEW MEXICO MANZANO MOUNTAINS, WEST OF BUILDINGS 4 AND T-59, LOOKING NORTHWEST - U. S. Veterans Administration Medical Center, 2100 Ridgecrest Southeast, Albuquerque, Bernalillo County, NM

  8. Silkworm protein: its possibility as an actuator

    NASA Astrophysics Data System (ADS)

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

    2006-03-01

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

  9. Antioxidant and anti-fatigue effects of anthocyanins of mulberry juice purification (MJP) and mulberry marc purification (MMP) from different varieties mulberry fruit in China.

    PubMed

    Jiang, Dong-Qing; Guo, Ying; Xu, Dian-Hong; Huang, Ya-Si; Yuan, Ke; Lv, Zhi-Qiang

    2013-09-01

    Anthocyanins, copiously distributed in a variety of colored fruits and vegetables, are probably the most important group of visible plant pigments besides chlorophyll. And the mulberry fruit is one of the anthocyanins-rich fruits. Total flavonols, total phenolic acids and anthocyanins contents of ten varieties mulberry juice purification (MJP) and mulberry marc purification (MMP) were determined. The highest content was 965.634.90 mg RE/g, 690.837.38 mg GAE/g and 272.001.20 mg cyanidin-3-glucoside/g FW, respectively. Moreover, MJP and MMP exhibited high antioxidant activity, including total force reduction (TRP), Fe? reducing power (FRAP) and DPPH radical scavenging capacity. In addition, the anti-fatigue activity of MJP and MMP was determined through mice-burden swimming experiments. Interestingly, the antioxidant and anti-fatigue capacities of MMP were much higher than those of MJP. The experimental results suggested that the generally discarded mulberry marc had greater value of development and utilization as food processing waste. PMID:23727333

  10. Transcriptome Analysis of Integument Differentially Expressed Genes in the Pigment Mutant (quail) during Molting of Silkworm, Bombyx mori

    PubMed Central

    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

  11. 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

  12. 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...

  13. [Mapping of Bmrbp1 gene in silkworm chromosome].

    PubMed

    Pan, Min Hui; Wang, Qiang; Shen, Yi Hong; Fu, Yong; Xu, Yuan; Lu, Cheng

    2007-10-01

    Mapping of silkworm Bmrbp1 gene in silkworm chromosome may lay the basis to investigate function of silkworm Bmrbp1 gene and to establish silkworm physical genetic map. In this study, FISH (fluorescence in situ hybridization) was carried out on meiotic chromosome using one sex regulation gene rbp1 of Bombyx mori as a probe. The experimental results demonstrated that: this gene was a single-copy gene of autosome, located at the site with relative length of 38.806 +/- 1.080% in the 5th chromosome of silkworm meiosis. PMID:18254343

  14. 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 β-alanine and N-β-alanyl dopamine (NBAD), but accumulation of dopamine. Following injection of β-alanine into bp mutants, the color pattern was reverted that of the wild-type silkworms. Additionally, melanic pupae resulting from knock-down of BmADC in the wild-type strain were obtained. These findings show that BmADC plays a crucial role in melanin metabolism and in the pigmentation pattern of the silkworm pupal stage. Finally, this study contributes to a better understanding of pupa pigmentation patterns in Lepidoptera. PMID:26077025

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

    PubMed Central

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

    2015-01-01

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

  16. 26. OVERALL VIEW, FROM SOUTH, OF MULBERRY STREET BRIDGE, SHOWING ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    26. OVERALL VIEW, FROM SOUTH, OF MULBERRY STREET BRIDGE, SHOWING BOTH SHEDS, MAIL CONVEYOR CONNECTING POST OFFICE BUILDING TO LOADING DOCK UNDER WEST SHED (RIGHT SIDE), PLATFORM CANOPIES AND CATENARY SYSTEM. NOTE STATE CAPITOL DOME VISIBLE BEYOND STATION BUILDING - Pennsylvania Railroad, Harrisburg Station & Trainshed, Market & South Fourth Streets, Harrisburg, Dauphin County, PA

  17. Mulberry and mulberry wine extract increase the number of mitochondria during brown adipogenesis.

    PubMed

    You, Yilin; Yuan, Xiaoxue; Lee, Hyuek Jong; Huang, Weidong; Jin, Wanzhu; Zhan, Jicheng

    2015-02-01

    Mulberry extract (ME) has been shown to possess beneficial effects towards obesity, but its mechanism is still unclear. In small mammals, mitochondria enriched brown adipose tissue (BAT) is known to convert protein's electrochemical energy to heat and maintain a constant body temperature. Improving the mitochondrial function or increasing the number of mitochondria could promote the metabolism of carbohydrate and fat. Thus, this study was designed to investigate the mitochondrial function regulated by ME and mulberry wine extract (MWE) during the brown adipogenesis. The C3H10T1/2 mesenchymal stem cell was treated with ME and MWE, both of which significantly (p < 0.05) increased the expression levels of fatty acid oxidation related genes such as peroxisome proliferator-activated receptor-γ coactivator-1α, PR domain-containing 16 and carnitine palmitoyltransferase 1α during brown adipogenesis. These changes were accompanied with increases in mitochondrial oxidative complex proteins upon ME and/or MWE exposure. Notably, ME and/or MWE also significantly (p < 0.05) increased the expression of the transcription factor A and the nuclear respiratory factor-1, which are the key transcription factors of mitochondrial biogenesis. In parallel, the mitochondrial copy number and brown adipose tissue specific gene-uncoupling protein-1 expression were dramatically (p < 0.05) elevated after ME or MWE treatment. Cyanidin-3-glucoside (Cy-3-glu) was found to be one of the most abundant anthocyanins in ME and MWE. Therefore, the BAT regulatory activity of ME and MWE might be, at least in part, due to the effect of Cy-3-glu. These results suggested that ME and MWE could ameliorate metabolic disease through an improvement in mitochondrial functions. PMID:25554947

  18. Molecular cloning and expression analyses of RPS3a gene from mulberry under abiotic stresses and among different mulberry varieties.

    PubMed

    Qian, J; Zhou, H; Zhao, M D; Wang, H; Li, F; Wang, Y H; Fang, R J; Zhao, W G; Kim, H J

    2016-01-01

    A full-length cDNA sequence coding ribosomal protein S3a of mulberry tree, which we designated MmRPS3a (GenBank accession No. KR610331), was cloned based on mulberry expressed sequence tags. Sequence analysis showed that the MmRPS3a is 1089 bp long and contains a 80-bp 5'-UTR (untranslated region) and a 220-bp 3'-UTR. Its open reading frame consists of a 789-bp encoding 262 amino acids with a predicted molecular weight of 30.053 kDa and an isoelectric point of 9.84. Homology analysis revealed that MmRPS3a gene is highly conservative in mulberry and other species including Morus notabilis, Theobroma cacao, and Ricinus communis. Phylogenetic analysis based on MmRPS3a of other species showed that mulberry had a closer relationship with Prunus persica, Arabidopsis thaliana, Solanum tuberosum, Solanum lycopersicum, and Vitis vinifera. The results of quantitative PCR analysis showed that the transcriptional level of MmRPS3a mRNA changed significantly under the conditions of hypothermia, aridity, salt stress, and varieties of differing resistances. PMID:27173298

  19. Effects of Mulberry Leaf Extract Rich in 1-Deoxynojirimycin on Blood Lipid Profiles in Humans

    PubMed Central

    Kojima, Yoshihiro; Kimura, Toshiyuki; Nakagawa, Kiyotaka; Asai, Akira; Hasumi, Keiji; Oikawa, Shinichi; Miyazawa, Teruo

    2010-01-01

    Mulberry leaves are rich in 1-deoxynojirimycin (DNJ), an inhibitor of ?-glucosidase. We previously showed that DNJ-rich mulberry leaf extract suppressed elevation of postprandial blood glucose in humans. The objective of this study was to evaluate the effects of DNJ-rich mulberry leaf extract on plasma lipid profiles in humans. An open-label, single-group study was conducted in 10 subjects with initial serum triglyceride (TG) level ?200mg/dl. Subjects ingested capsules containing DNJ-rich mulberry leaf extract at 12mg three times daily before meals for 12 weeks. Our findings showed a modest decrease in serum TG level and beneficial changes in the lipoprotein profile following 12-week administration of DNJ-rich mulberry leaf extract. No significant changes in hematological or biochemical parameters were observed during the study period; no adverse events associated with DNJ-rich mulberry leaf extract occurred. PMID:20838571

  20. Combination solar hothouse and silkworm breeding house

    SciTech Connect

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

    1980-01-01

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

  1. Neuropeptides of the silkworm, Bombyx mori.

    PubMed

    Nagasawa, H

    1992-05-15

    Six neuropeptides of the silkworm, Bombyx mori, have been isolated and chemically characterized during the past 10 years. They are bombyxin, prothoracicotropic hormone, pheromone-biosynthesis-activating neuropeptide/melanization-and-reddish-coloration hormone, diapause hormone, eclosion hormone, and adipokinetic hormone. Recent progress in research on these neuropeptides is described. PMID:1601106

  2. Chemical properties in fruits of mulberry species from the Xinjiang province of China.

    PubMed

    Jiang, Yan; Nie, Wen-Jing

    2015-05-01

    Mulberries are a widely cultivated foodstuff both in China and worldwide. However, there are stark differences in the nutritional values of mulberry species. To better appreciate these differences, we here describe the chemical characteristics of white (Morus alba L.), Russian (M. alba var. tatarica L.), and black (Morus nigra L.) mulberry fruits cultivated in the Xinjiang province of China. The chemical composition analysis was performed by official methods procedures. The amino acids were analysed by the phenyl isothiocyanate method. The 2,6-dichloroindophenol titrimetric method, the aluminium chloride colorimetric method, and the pH differential method were also used in measuring the content of reduced ascorbic acid, total flavonoids, and total monomeric anthocyanins, respectively. The black mulberry fruits had the highest content of reduced ascorbic acid (48.4 mg/100 g fw), titratable acidity (47.1 mg/g fw), and Fe (11.9 mg/100 g fw) of these 3 species. The Russian mulberry fruits had the highest EAA/TAA (essential amino acid/total amino acid) ratio at 44% followed by the white mulberry (42%) and the black mulberry (29%). The black mulberry fruits had found to be richest in terms of total flavonoids and total monomeric anthocyanins. These results are helpful for selecting mulberry species with abundant nutrients and phytochemicals for commercial cultivation. PMID:25529706

  3. Identification and characterization of Sox genes in the silkworm, Bombyx mori.

    PubMed

    Wei, Ling; Cheng, Daojun; Li, Dong; Meng, Meng; Peng, Lina; Tang, Lin; Pan, Minhui; Xiang, Zhonghuai; Xia, Qingyou; Lu, Cheng

    2011-06-01

    Sox genes encode a family of transcription factors with important roles in metazoan development, including sex-determination, embryogenesis, neurogenesis, and skeletogenesis. We identified Sox genes in the Bombyx mori genome and characterized their evolution and expression patterns. Nine Sox genes were annotated, and could be classified into five groups, B-F. Four Sox genes in the B group were tandemly clustered on one chromosome, a characteristic common to their orthologs in other insects. The intron number in the high-mobility group (HMG) box of Sox genes exhibited low diversity across surveyed insects. Based on 40 different silkworm variety genomes, we found a similar number of single nucleotide polymorphisms (SNPs) in the coding sequences of each Sox gene, for domesticated and wild groups. However, a gene-based examination showed that SoxB3 and SoxD might be evolving under positive selection during silkworm domestication. Phylogenetic analysis showed that SoxC, SoxD, and SoxF originated before the radiation of insects, and groups B and E evolved through gene duplication after the radiation of insects. Furthermore, BmSox21a, BmSoxB3, BmSoxD, and BmSoxE reveal stage, tissue, or sex-dependent expression patterns. These results provide a foundation for further surveying the functions of Sox genes during silkworm development and domestication. PMID:21161409

  4. Complete Genome Sequence of Mulberry Vein Banding Associated Virus, a New Tospovirus Infecting Mulberry

    PubMed Central

    Meng, Jiaorong; Liu, Pingping; Zhu, Liling; Zou, Chengwu; Li, Jieqiu; Chen, Baoshan

    2015-01-01

    Mulberry vein banding associated virus (MVBaV) that infects mulberry plants with typical vein banding symptoms had been identified as a tentative species of the genus Tospovirus based on the homology of N gene sequence to those of tospoviruses. In this study, the complete sequence of the tripartite RNA genome of MVBaV was determined and analyzed. The L RNA has 8905 nucleotides (nt) and encodes the putative RNA-dependent RNA polymerase (RdRp) of 2877 aa amino acids (aa) in the viral complementary (vc) strand. The RdRp of MVBaV shares the highest aa sequence identity (85.9%) with that of Watermelon silver mottle virus (WSMoV), and contains conserved motifs shared with those of the species of the genus Tospovirus. The M RNA contains 4731 nt and codes in ambisense arrangement for the NSm protein of 309 aa in the sense strand and the Gn/Gc glycoprotein precursor (GP) of 1,124 aa in the vc strand. The NSm and GP of MVBaV share the highest aa sequence identities with those of Capsicum chlorosis virus (CaCV) and Groundnut bud necrosis virus (GBNV) (83.2% and 84.3%, respectively). The S RNA is 3294 nt in length and contains two open reading frames (ORFs) in an ambisense coding strategy, encoding a 439-aa non-structural protein (NSs) and the 277-aa nucleocapsid protein (N), respectively. The NSs and N also share the highest aa sequence identity (71.1% and 74.4%, respectively) with those of CaCV. Phylogenetic analysis of the RdRp, NSm, GP, NSs, and N proteins showed that MVBaV is most closely related to CaCV and GBNV and that these proteins cluster with those of the WSMoV serogroup, and that MVBaV seems to be a species bridging the two subgroups within the WSMoV serogroup of tospoviruses in evolutionary aspect, suggesting that MVBaV represents a distinct tospovirus. Analysis of S RNA sequence uncovered the highly conserved 5’-/3’-ends and the coding regions, and the variable region of IGR with divergent patterns among MVBaV isolates. PMID:26291718

  5. Mulberry leaf phenolics ameliorate hyperglycemia-induced oxidative stress and stabilize mitochondrial membrane potential in HepG2 cells.

    PubMed

    Zou, Yu-Xiao; Shen, Wei-Zhi; Liao, Sen-Tai; Liu, Fan; Zheng, Shan-Qing; Blumberg, Jeffrey B; Chen, C-Y Oliver

    2014-12-01

    To investigate the effect of phenolics in mulberry leaves (mulberry leaf phenolics; MLP) on hyperglycemia-induced oxidative stress and mitochondrial membrane potential (ΔΨm) in HepG2 cells; we treated HepG2 with glucose [5.5 (N-Glc) or 50 mmol/L (Hi-Glc)] with or without MLP at 10 or 100 µmol/L gallic acid equivalents and assessed level of reactive oxidant species (ROS), ΔΨm, malondialdehyde (MDA) and nuclear factor-kappaB (NF-κB) activation. Hi-Glc-induced oxidative damage was demonstrated by a series of increase in superoxides (560%, 0.5 h), MDA (400%, 24 h), NF-κB activation (474%, 4 h) and a wild fluctuation of ΔΨm relative to the control cells (p ≤ 0.05). MLP treatments ameliorate Hi-Glc-induced negative effects by a 40% reduction in ROS production, 34-44% reduction in MDA production, over 35% inhibition of NF-κB activation, as well as exert protective effect on HepG2 cells from change in ΔΨm. Our data show that MLP in vitro can protect hepatoctyes from hyperglycemia-induced oxidative damages. PMID:25026091

  6. The effects of juice processing on black mulberry antioxidants.

    PubMed

    Tomas, Merve; Toydemir, Gamze; Boyacioglu, Dilek; Hall, Robert; Beekwilder, Jules; Capanoglu, Esra

    2015-11-01

    Black mulberry fruit is processed to juice at significant scale in Turkey. The effect of industrial-scale juice production on black mulberry antioxidants was evaluated using samples collected from the main steps of processing; including the selection of fruits, washing, mechanical milling, mashing, cold pressing, pasteurization, and filling-packing. Two major anthocyanins (cyanidin-3-glucoside and cyanidin-3-rutinoside), two phenolic acids (3- and caffeoylquinic acid) and 3 flavonols (rutin, quercetin-3-glucoside, and quercetin-malonyl-glucoside) were identified using LC-QTOF-MS and were quantified using HPLC. Approximately, 60-70% of the fruit anthocyanins were retained in the final juice, which also contained high levels of caffeoylquinic acids, relative to the fruit. Mashing and pressing were the steps which were effective for the recovery of fruit polyphenolics into the juice fraction. Moreover, an in vitro gastrointestinal digestion model, applied to determine the effect of processing on the bioavailability of mulberry antioxidants, indicated a higher anthocyanin bioavailability for the fruit matrix than for the juice matrix. PMID:25976822

  7. Genome Sequence of a Xylella fastidiosa Strain Causing Mulberry Leaf Scorch Disease in Maryland.

    PubMed

    Guan, Wei; Shao, Jonathan; Zhao, Tingchang; Huang, Qi

    2014-01-01

    Xylella fastidiosa causes bacterial leaf scorch in landscape trees, including mulberry. We determined the draft genome of the mulberry strain Mul-MD in order to gain a better understanding of the molecular basis of strain divergence, host specificity, nutrient requirements, and pathogenicity, as well as to develop genome-based specific detection methods. PMID:24604658

  8. Enterobacter morus sp. nov., a novel Enterobacter species associated with bacterial wilt on mulberry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A mulberry pathogenetic bacterial strain R18-2T isolated from the diseased mulberry root was analyzed by a polyphasic taxonomic study. Comparative 16S rRNA gene sequence analysis combined with rpoB gene sequence analysis allocated the strain R18-2T to the genus Enterobacter. The strain was Gram nega...

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

    NASA Astrophysics Data System (ADS)

    Wyman, Aaron J.; Webb, Mary Alice

    2007-04-01

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

  10. Rearing of silkworm under hypobaric and hypoxia conditions

    NASA Astrophysics Data System (ADS)

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

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

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

    PubMed

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

    2010-01-01

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

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

    PubMed

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

    2016-01-01

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

  13. Molecular mapping and characterization of the silkworm apodal mutant

    PubMed Central

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

    2016-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2015-07-01

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

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

    PubMed Central

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

    2015-01-01

    The thermoprotective mechanisms of insects remain largely unknown. We reported the Bombyx mori contractile (cot) behavioral mutant with thermo-sensitive seizures phenotype. At elevated temperatures, the cot mutant exhibit seizures associated with strong contractions, rolling, vomiting, and a temporary lack of movement. We narrowed a region containing cot to ~268 kb by positional cloning and identified the mutant gene as Bmsei which encoded a potassium channel protein. Bmsei was present in both the cell membrane and cytoplasm in wild-type ganglia but faint in cot. Furthermore, Bmsei was markedly decreased upon high temperature treatment in cot mutant. With the RNAi method and injecting potassium channel blockers, the wild type silkworm was induced the cot phenotype. These results demonstrated that Bmsei was responsible for the cot mutant phenotype and played an important role in thermoprotection in silkworm. Meanwhile, comparative proteomic approach was used to investigate the proteomic differences. The results showed that the protein of Hsp-1 and Tn1 were significantly decreased and increased on protein level in cot mutant after thermo-stimulus, respectively. Our data provide insights into the mechanism of thermoprotection in insect. As cot phenotype closely resembles human epilepsy, cot might be a potential model for the mechanism of epilepsy in future. PMID:26198671

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

    PubMed

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

    2015-01-01

    The thermoprotective mechanisms of insects remain largely unknown. We reported the Bombyx mori contractile (cot) behavioral mutant with thermo-sensitive seizures phenotype. At elevated temperatures, the cot mutant exhibit seizures associated with strong contractions, rolling, vomiting, and a temporary lack of movement. We narrowed a region containing cot to ~268 kb by positional cloning and identified the mutant gene as Bmsei which encoded a potassium channel protein. Bmsei was present in both the cell membrane and cytoplasm in wild-type ganglia but faint in cot. Furthermore, Bmsei was markedly decreased upon high temperature treatment in cot mutant. With the RNAi method and injecting potassium channel blockers, the wild type silkworm was induced the cot phenotype. These results demonstrated that Bmsei was responsible for the cot mutant phenotype and played an important role in thermoprotection in silkworm. Meanwhile, comparative proteomic approach was used to investigate the proteomic differences. The results showed that the protein of Hsp-1 and Tn1 were significantly decreased and increased on protein level in cot mutant after thermo-stimulus, respectively. Our data provide insights into the mechanism of thermoprotection in insect. As cot phenotype closely resembles human epilepsy, cot might be a potential model for the mechanism of epilepsy in future. PMID:26198671

  17. Mulberry Leaf Reduces Oxidation and C-Reactive Protein Level in Patients with Mild Dyslipidemia

    PubMed Central

    Aramwit, Pornanong; Supasyndh, Ouppatham; Siritienthong, Tippawan; Bang, Nipaporn

    2013-01-01

    C-reactive protein (CRP) is the inflammatory marker that could represent the inflammation in blood vessels resulted from dyslipidemia. The objective of this study was to evaluate the antioxidative activity of mulberry leaf powder using DPPH assay and the effect of mulberry leaf powder on lipid profile, CRP level, and antioxidative parameters in mild dyslipidemia patients. A within-subjects design was conducted and patients received three tablets of 280 mg mulberry leaf powder three times a day before meals for 12 weeks. Total of 25 patients were enrolled but one subject was excluded. After three months of mulberry leaf consumption, serum triglyceride and low-density lipoprotein (LDL) level were significantly reduced and more than half of all patients' CRP levels decreased every month as well as the mean CRP level but no statistically significant difference was found. The average erythrocyte glutathione peroxidase activity of patients was increased but not at significant level; however, the mean serum 8-isoprostane level was significantly lower after mulberry treatment for 12 weeks. It can be concluded that mulberry leaf powder exhibited antioxidant activity and mulberry leaf powder has potential to decrease serum triglyceride, LDL, and CRP levels in mild dyslipidemia patients without causing severe adverse reactions. PMID:23484158

  18. Reducing soil erosion and nutrient loss on sloping land under crop-mulberry management system.

    PubMed

    Fan, Fangling; Xie, Deti; Wei, Chaofu; Ni, Jiupai; Yang, John; Tang, Zhenya; Zhou, Chuan

    2015-09-01

    Sloping croplands could result in soil erosion, which leads to non-point source pollution of the aquatic system in the Three Gorges Reservoir Region. Mulberry, a commonly grown cash plant in the region, is traditionally planted in contour hedgerows as an effective management practice to control soil erosion and non-point source pollution. In this field study, surface runoff and soil N and P loss on sloping land under crop-mulberry management were investigated. The experiments consisted of six crop-mulberry treatments: Control (no mulberry hedgerow with mustard-corn rotation); T1 (two-row contour mulberry with mustard-corn rotation); T2 (three-row contour mulberry with mustard-corn rotation); T3 (border mulberry and one-row contour mulberry with mustard-corn rotation); T4 (border mulberry with mustard-corn rotation); T5 (two-row longitudinal mulberry with mustard). The results indicated that crop-mulberry systems could effectively reduce surface runoff and soil and nutrient loss from arable slope land. Surface runoff from T1 (342.13 m(3) hm(-2)), T2 (260.6 m(3) hm(-2)), T3 (113.13 m(3) hm(-2)), T4 (114 m(3) hm(-2)), and T5 (129 m(3) hm(-2)) was reduced by 15.4, 35.6, 72.0, 71.8, and 68.1%, respectively, while soil loss from T1 (0.21 t hm(-2)), T2 (0.13 t hm(-2)), T3 (0.08 t hm(-2)), T4 (0.11 t hm(-2)), and T5 (0.12 t hm(-2)) was reduced by 52.3, 70.5, 81.8, 75.0, and 72.7%, respectively, as compared with the control. Crop-mulberry ecosystem would also elevate soil N by 22.3% and soil P by 57.4%, and soil nutrient status was contour-line dependent. PMID:25957753

  19. Non-mulberry Silk Fibroin Biomaterial for Corneal Regeneration

    PubMed Central

    Hazra, Sarbani; Nandi, Sudip; Naskar, Deboki; Guha, Rajdeep; Chowdhury, Sushovan; Pradhan, Nirparaj; Kundu, Subhas C.; Konar, Aditya

    2016-01-01

    Purpose: Successful repair of a damaged corneal surface is a great challenge and may require the use of a scaffold that supports cell growth and differentiation. Amniotic membrane is currently used for this purpose, in spite of its limitations. A thin transparent silk fibroin film from non-mulberry Antheraea mylitta (Am) has been developed which offers to be a promising alternative. The silk scaffolds provide sufficient rigidity for easy handling, the scaffolds support the sprouting, migration, attachment and growth of epithelial cells and keratocytes from rat corneal explants; the cells form a cell sheet, preserve their phenotypes, express cytokeratin3 and vimentin respectively. The films also support growth of limbal stem cell evidenced by expression of ABCG2. The cell growth on the silk film and the amniotic membrane is comparable. The implanted film within the rabbit cornea remains transparent, stable. The clinical examination as well as histology shows absence of any inflammatory response or neovascularization. The corneal surface integrity is maintained; tear formation, intraocular pressure and electroretinography of implanted eyes show no adverse changes. The silk fibroin film from non-mulberry silk worms may be a worthy candidate for use as a corneal scaffold. PMID:26908015

  20. DISPOSITION OF MULBERRY POLLEN IN THE HUMAN RESPIRATORY SYSTEMS: A MATHEMATICAL MODEL

    EPA Science Inventory

    Inhaled particle deposition sites must be identified to effectively treat human airway diseases. e have determined distribution patterns of a selected aeroallergen, mulberry pollen, among human extrathoracic (ET: i.e., oronasopharyngeal) regions and the lung. redictive model vali...

  1. Quantitative determination of 1-deoxynojirimycin in mulberry leaves using liquid chromatography-tandem mass spectrometry.

    PubMed

    Nuengchamnong, Nitra; Ingkaninan, Kornkanok; Kaewruang, Wiroje; Wongareonwanakij, Sathaporn; Hongthongdaeng, Bhinai

    2007-08-15

    A novel HPLC-MS/MS method was developed for the quantitative determination of 1-deoxynojirimycin (DNJ), a potent glucosidase inhibitor present in mulberry leaves (Morus alba L.). DNJ was isolated from the mulberry leave extract on a TSKgel Amide-80 column using a mixture of 0.1% formic acid and acetonitrile as a mobile phase at a flow rate of 0.6 ml/min. A triple quadrupole mass spectrometry using electrospray ionization source in a positive ion mode under multiple reaction monitoring with the [M+H]+ ions, m/z 164.4/109.9 was used. The detection limit (S/N=3) was 75 pg and quantitation limit (S/N=10) was 100 pg. The comparison of mulberry leaves of different ages showed that the DNJ level was higher in mulberry shoots than young and mature leaves. PMID:17512690

  2. Optimization of Ultrasound-Assisted Extraction of Anthocyanins from Mulberry, Using Response Surface Methodology

    PubMed Central

    Zou, Tang-Bin; Wang, Min; Gan, Ren-You; Ling, Wen-Hua

    2011-01-01

    Mulberry is one of the most widely used traditional Chinese medicines. Anthocyanins are the main bioactive components of mulberry, and possess important biological activities, such as antimicrobial, anti-inflammatory and antioxidant activities. This study investigated the ultrasound-assisted extraction (UAE) of anthocyanins from mulberry by using response surface methodology (RSM). The extraction conditions associated with anthocyanin yield, including extraction solvent, liquid-to-solid rate, temperature and extraction time, are discussed. The optimal conditions obtained by RSM for UAE from mulberry include 63.8% methanol contains 1% (v/v) trifluoroacetic acid (TFA), 43.2 °C temperature, 23.8 (v/w) liquid-to-solid ratio, and 40 min time for the maximum yield (64.70 ± 0.45 mg/g). The results indicated that the UAE can be an effective method for the extraction of some active components from plant materials. PMID:21686165

  3. 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

  4. 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

  5. Mulberry Transcription Factor MnDREB4A Confers Tolerance to Multiple Abiotic Stresses in Transgenic Tobacco

    PubMed Central

    Guo, Qing; Zhang, Meng; Cao, Bo-Ning; Xiang, Zhong-Huai; Zhao, Ai-Chun

    2015-01-01

    The dehydration responsive element binding (DREB) transcription factors have been reported to be involved in stress responses. Most studies have focused on DREB genes in subgroups A-1 and A-2 in herbaceous plants, but there have been few reports on the functions of DREBs from the A-3–A-6 subgroups and in woody plants. Moreover, mulberry trees are ecologically and economically important perennial woody plants, but there has been little research on its stress physiology, biochemistry and molecular biology. In this study, a DREB gene from the mulberry tree, designated as MnDREB4A, classified into the A-4 subgroup by our previous study, was selected for further characterization. Our results showed that the MnDREB4A protein was localized to the nucleus where it activated transcription. The promoter of MnDREB4A can direct prominent expression downstream of the β-glucuronidase (GUS) gene under heat, cold, drought and salt stress, and GUS staining was deepest after 12 h of stress treatment. The MnDREB4A-overexpression transgenic tobacco showed the improved growth phenotype under untreated conditions, such as greener leaves, longer roots, and lower water loss and senescence rates. Overexpression of MnDREB4A in tobacco can significantly enhance tolerance to heat, cold, drought, and salt stresses in transgenic plants. The leaf discs and seedlings of transgenic plants reduced leaf wilting and senescence rates compared to the wild type plants under the different stress conditions. Further investigation showed that transgenic plants also had higher water contents and proline contents, and lower malondialdehyde contents under untreated condition and stress conditions. Our results indicate that the MnDREB4A protein plays an important role in plant stress tolerance. PMID:26695076

  6. Mulberry Transcription Factor MnDREB4A Confers Tolerance to Multiple Abiotic Stresses in Transgenic Tobacco.

    PubMed

    Liu, Xue-Qin; Liu, Chang-Ying; Guo, Qing; Zhang, Meng; Cao, Bo-Ning; Xiang, Zhong-Huai; Zhao, Ai-Chun

    2015-01-01

    The dehydration responsive element binding (DREB) transcription factors have been reported to be involved in stress responses. Most studies have focused on DREB genes in subgroups A-1 and A-2 in herbaceous plants, but there have been few reports on the functions of DREBs from the A-3-A-6 subgroups and in woody plants. Moreover, mulberry trees are ecologically and economically important perennial woody plants, but there has been little research on its stress physiology, biochemistry and molecular biology. In this study, a DREB gene from the mulberry tree, designated as MnDREB4A, classified into the A-4 subgroup by our previous study, was selected for further characterization. Our results showed that the MnDREB4A protein was localized to the nucleus where it activated transcription. The promoter of MnDREB4A can direct prominent expression downstream of the β-glucuronidase (GUS) gene under heat, cold, drought and salt stress, and GUS staining was deepest after 12 h of stress treatment. The MnDREB4A-overexpression transgenic tobacco showed the improved growth phenotype under untreated conditions, such as greener leaves, longer roots, and lower water loss and senescence rates. Overexpression of MnDREB4A in tobacco can significantly enhance tolerance to heat, cold, drought, and salt stresses in transgenic plants. The leaf discs and seedlings of transgenic plants reduced leaf wilting and senescence rates compared to the wild type plants under the different stress conditions. Further investigation showed that transgenic plants also had higher water contents and proline contents, and lower malondialdehyde contents under untreated condition and stress conditions. Our results indicate that the MnDREB4A protein plays an important role in plant stress tolerance. PMID:26695076

  7. Analysis of Functional Constituents in Mulberry (Morus alba L.) Twigs by Different Cultivars, Producing Areas, and Heat Processings

    PubMed Central

    Choi, Sang Won; Jang, Yeon Jeong; Lee, Yu Jin; Leem, Hyun Hee; Kim, Eun Ok

    2013-01-01

    Four functional constituents, oxyresveratrol 3′-O-β-D-glucoside (ORTG), oxyresveratrol (ORT), t-resveratrol (RT), and moracin (MC) were isolated from the ethanolic extract of mulberry (Morus alba L.) twigs by a series of isolation procedures, including solvent fractionation, and silica-gel, ODS-A, and Sephadex LH-20 column chromatographies. Their chemical structures were identified by NMR and FABMS spectral analysis. Quantitative changes of four phytochemicals in mulberry twigs were determined by HPLC according to cultivar, producing area, and heat processing. ORTG was a major abundant compound in the mulberry twigs, and its levels ranged from 23.7 to 105.5 mg% in six different mulberry cultivars. Three other compounds were present in trace amounts (<1 mg/100 g) or were not detected. Among mulberry cultivars examined, “Yongcheon” showed the highest level of ORTG, whereas “Somok” had the least ORTG content. Levels of four phytochemicals in the mulberry twigs harvested in early September were higher than those harvested in early July. Levels of ORTG and ORT in the “Cheongil” mulberry twigs produced in the Uljin area were higher than those produced in other areas. Generally, levels of ORTG and ORT in mulberry twigs decreased with heat processing, such as steaming, and microwaving except roasting, whereas those of RT and MC did not considerably vary according to heat processing. These results suggest that the roasted mulberry twigs may be useful as potential sources of functional ingredients and foods. PMID:24551827

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

    PubMed

    Ishii, Masaki; Matsumoto, Yasuhiko; Sekimizu, Kazuhisa

    2016-01-01

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

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

    PubMed

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

    2016-01-15

    With the recent availability of the genomes of many viruses and the silkworm, Bombyx mori, as well as a variety of Basic Local Alignment Search Tool (BLAST) programs, a new opportunity to gain insight into the interaction of viruses with the silkworm is possible. This study aims to determine the possible existence of sequence identities between the genomes of viruses and the silkworm and attempts to explain this phenomenon. BLAST searches of the genomes of viruses against the silkworm genome were performed using the resources of the National Center for Biotechnology Information. All studied viruses contained variable numbers of short regions with sequence identity to the genome of the silkworm. The short regions of sequence identity in the genome of the silkworm may be derived from the genomes of viruses in the long history of silkworm-virus interaction. This study is the first to compare these genomes, and may contribute to research on the interaction between viruses and the silkworm. PMID:26432002

  10. 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

  11. 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

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

    PubMed Central

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

    2015-01-01

    The silkworm Dominant trimolting (Moltinism, M3) mutant undergoes three larval molts and exhibits precocious metamorphosis. In this study, we found that compared with the wild-type (WT) that undergoes four larval molts, both the juvenile hormone (JH) concentration and the expression of the JH-responsive gene Krüppel homolog 1 (Kr-h1) began to be greater in the second instar of the M3 mutant. A positional cloning analysis revealed that only the homeodomain transcription factor gene Sex combs reduced (Scr) is located in the genomic region that is tightly linked to the M3 locus. The expression level of the Scr gene in the brain-corpora cardiaca-corpora allata (Br-CC-CA) complex, which controls the synthesis of JH, was very low in the final larval instar of both the M3 and WT larvae, and exhibited a positive correlation with JH titer changes. Importantly, luciferase reporter analysis and electrophoretic mobility shift assay (EMSA) demonstrated that the Scr protein could promote the transcription of genes involved in JH biosynthesis by directly binding to the cis-regulatory elements (CREs) of homeodomain protein on their promoters. These results conclude that the homeodomain protein Scr is transcriptionally involved in the regulation of JH biosynthesis in the silkworm. PMID:26540044

  13. 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

  14. The Mulberry Cogeneration Facility: Design features and operating success

    SciTech Connect

    Jasper, W.M.; Wierschem, R.A.; Gray, D.C.

    1995-12-31

    The Mulberry Cogeneration Facility is in successful, reliable, and profitable service, with the complex requirements of zero discharge, extensive water treatment, process steam supply, and inlet chillers having been reduced to routine, automated operation. What was initially visualized by the developer as a straightforward combined cycle plant became much more than that as the various requirements for environmental and regulatory compliance became necessities, along with the desire to maximize output and revenues in response to growing power demands. A delicate balance exists between a financially successful cogeneration project and the Owners` obligation to the community and the environment. As competitive pressures grow and expectations of investors, insurers, operators, and the public increase, still more creativity will be required to provide a safe, reliable, cost-effective plant on ever shorter schedules.

  15. Microwave-assisted extraction of polysaccharides from mulberry leaves.

    PubMed

    Thirugnanasambandham, K; Sivakumar, V; Maran, J Prakash

    2015-01-01

    In this study, microwave-assisted extraction (MAE) of polysaccharides from mulberry leaves was investigated using response surface methodology (RSM). The effects of three extraction factors on the yield of polysaccharides was examined. The results showed that optimum extraction conditions were determined as follows: weight of the sample of 20 g, microwave power of 170 W, extraction time of 10 min. Under these optimal extraction conditions, polysaccharide yield was found to be 9.41%. Three factors-three level Box-Behnken response surface design (BBD) coupled with RSM was used to model the extraction process. ANOVA was used to examine the statistical significance of the developed model. Extracted polysaccharide was analyzed using Fourier transform infrared (FT-IR) spectroscopy. PMID:25064558

  16. 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 AlCl₃ 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

  17. Effect of gossypol and gossypol related compounds on mulberry pyralid (diaphania pyloalis walker, lepidoptera: pyralidae), a pest of the Mulberry Tree

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Gossypol, gossypurpurin and diaminogossypol were tested for inhibitory effects against feeding mulberry pyralid larvae (Diaphania pyloalis Walker, Lepidoptera: Pyralidae). The inclusion of very low concentrations of these compounds (10, 50 or 100 µmoles/g) in artificial diets increased the number of...

  18. 33 CFR 334.280 - James River between the entrance to Skiffes Creek and Mulberry Point, Va.; army training and...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 33 Navigation and Navigable Waters 3 2012-07-01 2012-07-01 false James River between the entrance to Skiffes Creek and Mulberry Point, Va.; army training and small craft testing area. 334.280 Section... Creek and Mulberry Point, Va.; army training and small craft testing area. (a) The restricted...

  19. 33 CFR 334.280 - James River between the entrance to Skiffes Creek and Mulberry Point, Va.; army training and...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 3 2011-07-01 2011-07-01 false James River between the entrance to Skiffes Creek and Mulberry Point, Va.; army training and small craft testing area. 334.280 Section... Creek and Mulberry Point, Va.; army training and small craft testing area. (a) The restricted...

  20. 33 CFR 334.280 - James River between the entrance to Skiffes Creek and Mulberry Point, Va.; army training and...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 33 Navigation and Navigable Waters 3 2013-07-01 2013-07-01 false James River between the entrance to Skiffes Creek and Mulberry Point, Va.; army training and small craft testing area. 334.280 Section... Creek and Mulberry Point, Va.; army training and small craft testing area. (a) The restricted...

  1. 33 CFR 334.280 - James River between the entrance to Skiffes Creek and Mulberry Point, Va.; army training and...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 33 Navigation and Navigable Waters 3 2014-07-01 2014-07-01 false James River between the entrance to Skiffes Creek and Mulberry Point, Va.; army training and small craft testing area. 334.280 Section... Creek and Mulberry Point, Va.; army training and small craft testing area. (a) The restricted...

  2. 33 CFR 334.280 - James River between the entrance to Skiffes Creek and Mulberry Point, Va.; army training and...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false James River between the entrance to Skiffes Creek and Mulberry Point, Va.; army training and small craft testing area. 334.280 Section... Creek and Mulberry Point, Va.; army training and small craft testing area. (a) The restricted...

  3. Host status and fruit odor response of Drosophila suzukii (Diptera: Drosophilidae) to figs and mulberries.

    PubMed

    Yu, Doris; Zalom, F G; Hamby, K A

    2013-08-01

    Drosophila suzukii Matsumura (Diptera: Drosophilidae) is an agricultural pest with a wide host range. It is known to infest fruit that are still ripening on the plant, as well as rotting and damaged fruit. Our study sought to determine whether D. suzukii use mulberries (Morus spp.) and figs (Ficus carica (L.)) as hosts, as their host status was ambiguous. Accordingly, we collected 25 field-infested fruit and counted the numbers of D. suzukii emerging from them. We also sought to determine whether female D. suzukii would respond to olfactory cues from ripe figs and mulberries. As the host population has been known to impact host odor response, flies from mulberry, fig, and cherry origins were tested in "one-choice" olfactometry studies. Our results show that mulberries and figs can serve as hosts for D. suzukii and that female flies will respond to their odors. The host population did affect response to fruit odors, although further studies are necessary to determine habitat fidelity. This has implications for management of this pest, especially in backyard and mixed fruit orchard situations, which commonly occur in the current range of D. suzukii, and fig and mulberry may serve as a pest reservoir for other hosts and cultivated crops. PMID:24020313

  4. Phenotypic and Transcriptomic Analyses of Autotetraploid and Diploid Mulberry (Morus alba L.)

    PubMed Central

    Dai, Fanwei; Wang, Zhenjiang; Luo, Guoqing; Tang, Cuiming

    2015-01-01

    Autopolyploid plants and their organs are often larger than their diploid counterparts, which makes them attractive to plant breeders. Mulberry (Morus alba L.) is an important commercial woody plant in many tropical and subtropical areas. In this study, we obtained a series of autotetraploid mulberry plants resulting from a colchicine treatment. To evaluate the effects of genome duplications in mulberry, we compared the phenotypes and transcriptomes of autotetraploid and diploid mulberry trees. In the autotetraploids, the height, breast-height diameter, leaf size, and fruit size were larger than those of diploids. Transcriptome data revealed that of 21,229 expressed genes only 609 (2.87%) were differentially expressed between diploids and autotetraploids. Among them, 30 genes were associated with the biosynthesis and signal transduction of plant hormones, including cytokinin, gibberellins, ethylene, and auxin. In addition, 41 differentially expressed genes were involved in photosynthesis. These results enhance our understanding of the variations that occur in mulberry autotetraploids and will benefit future breeding work. PMID:26402678

  5. Agrobacterium-mediated transient MaFT expression in mulberry (Morus alba L.) leaves.

    PubMed

    Wu, Su-Li; Yang, Xiao-Bing; Liu, Li-Qun; Jiang, Tao; Wu, Hai; Su, Chao; Qian, Yong-Hua; Jiao, Feng

    2015-01-01

    To optimize Agrobacterium-mediated transient transformation assay in mulberry (Morus alba L.), various infiltration methods, Agrobacterium tumefaciens (A. tumefaciens) strains, and bacterial concentrations were tested in mulberry seedlings. Compared with LBA4404, GV3101 harboring pBE2133 plasmids presented stronger GUS signals at 3 days post infiltration using syringe. Recombinant plasmids pBE2133:GFP and pBE2133:GFP:MaFT were successfully constructed. Transient expression of MaFT:GFP protein was found in leaves, petiole (cross section), and shoot apical meristem (SAM) of mulberry according to the GFP signal. Moreover, MaFT:GFP mRNA was also detected in leaves and SAM via RT-PCR and qRT-PCR. An efficient transient transformation system could be achieved in mulberry seedlings by syringe using A. tumefaciens GV3101 at the OD600 of 0.5. The movement of MaFT expression from leaves to SAM might trigger the precocious flowering of mulberry. PMID:26024368

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

    PubMed

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

    2015-12-10

    Intestinal immune response is a front line of host defense. The host factors that participate in intestinal immunity response remain largely unknown. We recently reported that Translationally Controlled Tumor Protein (BmTCTP) was obtained by constructing a phage display cDNA library of the silkworm midgut and carrying out high throughput screening of pathogen binding molecules. To further address the function of BmTCTP in silkworm intestinal immunity, transgenic RNAi silkworms were constructed by microinjection piggBac plasmid to Dazao embryos. The antimicrobial capacity of transgenic silkworm decreased since the expression of gut antimicrobial peptide from transgenic silkworm was not sufficiently induced during oral microbial challenge. Moreover, dynamic ERK phosphorylation from transgenic silkworm midgut was disrupted. Taken together, the innate immunity of intestinal was suppressed through disruption of dynamic ERK phosphorylation after oral microbial infection as a result of RNAi-mediated knockdown of midgut TCTP in transgenic silkworm. PMID:26302749

  7. Effect of ultra-high pressure homogenisation processing on phenolic compounds, antioxidant capacity and anti-glucosidase of mulberry juice.

    PubMed

    Yu, Yuanshan; Xu, Yujuan; Wu, Jijun; Xiao, Gengsheng; Fu, Mangqin; Zhang, Yousheng

    2014-06-15

    In this study, the effects of ultra-high pressure homogenisation (UHPH) processing at 200 MPa for 1-3 successive passes (inlet temperatures at 4°C) were compared with pasteurisation (95°C, 1 min) processing on phenolic compounds, antioxidant capacity (ORAC value) and anti-glucosidase of mulberry juice. Compared with thermal pasteurisation processing, the more reductions in the anthocyanins, phenolic acids (gallic, protocatechuic, caffeic and p-coumaric acids, and a unknown hydroxycinnamic acid) and quercetin aglycone contents, as well as ORAC value were observed during UHPH processing of mulberry juice, whereas all reductions above during UHPH processing could be inhibited by adding ascorbic acid to mulberry juice. Besides, no significant change (p>0.05) in the α-glucosidase inhibitory activity was observed during UHPH processing of mulberry juice, but showed a 14% reduction in mulberry juice processed by thermal pasteurisation. PMID:24491708

  8. Sex determination: insights from the silkworm.

    PubMed

    Suzuki, Masataka G

    2010-09-01

    The sex-determining system differs considerably among organisms. Even among insect species, the genetic system for sexdetermination is highly diversified. In Drosophila melanogaster, somatic sexual differentiation is regulated by a well characterized genetic hierarchy X:A > Sxl > tra/tra2 > dsx and fru. This cascade seems to control sex determination in all Drosophila species and is partially conserved in another dipteran species, Ceratitis capitata: Cctra/Cctra-2 > Ccdsx and Ccfru. However, in the silkworm, Bombyx mori, femaleness is determined by the presence of a dominant feminizing factor on the W chromosome. Moreover, no sex-specific regulatory Sxl homolog has been isolated from B. mori. Also, no tra homolog has yet been found in the Bombyx genome. Despite such differences, dsx homolog of B. mori (Bmdsx) is implicated in the sex determination. Bmdsx produces alternatively spliced mRNA isoforms that encode sex specific transcription factors as observed in dsx. While the female-specific splicing of dsx is activated by splicing activators, Tra and Tra2, the female splicing of Bmdsx represents the default mode. Instead, a splicing inhibitor, BmPSI is involved in the regulation of male-specific splicing of Bmdsx. Since BmPSI does not exhibit any sequence relationship to known SR proteins, such as Tra and Tra2, the regulatory mechanism of sex-specific alternative splicing of Bmdsx is distinct from that of dsx. PMID:20877002

  9. Fatty acids composition of Spanish black (Morus nigra L.) and white (Morus alba L.) mulberries.

    PubMed

    Sánchez-Salcedo, Eva M; Sendra, Esther; Carbonell-Barrachina, Ángel A; Martínez, Juan José; Hernández, Francisca

    2016-01-01

    This research has determined qualitatively and quantitatively the fatty acids composition of white (Morus alba) and black (Morus nigra) fruits grown in Spain, in 2013 and 2014. Four clones of each species were studied. Fourteen fatty acids were identified and quantified in mulberry fruits. The most abundant fatty acids were linoleic (C18:2), palmitic (C16:0), oleic (C18:1), and stearic (C18:0) acids in both species. The main fatty acid in all clones was linoleic (C18:2), that ranged from 69.66% (MN2) to 78.02% (MA1) of the total fatty acid content; consequently Spanish mulberry fruits were found to be rich in linoleic acid, which is an essential fatty acid. The fatty acid composition of mulberries highlights the nutritional and health benefits of their consumption. PMID:26213011

  10. Identification and characterization of Lateral Organ Boundaries Domain genes in mulberry, Morus notabilis.

    PubMed

    Luo, Yiwei; Ma, Bi; Zeng, Qiwei; Xiang, Zhonghuai; He, Ningjia

    2016-06-01

    Genes from the plant specific Lateral Organ Boundaries Domain (LBD) family encode transcriptional regulators that have a variety of functions in various physiological and developmental processes. In the present study, 31 LBD genes were identified in the mulberry genome. The genome features of all MnLBD genes and phylogenetic studies with Arabidopsis LBD protein sequences, accompanied by the expression analysis of each of the Morus LBD genes provide insights into the functional prediction of mulberry LBDs. The genome-wide surveys of the current mulberry genome have resulted in the identification of catalogs of MnLBD genes that may function in the development of leaf, root, and secondary metabolism in Morus sp. PMID:27014591

  11. Identification and characterization of Lateral Organ Boundaries Domain genes in mulberry, Morus notabilis

    PubMed Central

    Luo, Yiwei; Ma, Bi; Zeng, Qiwei; Xiang, Zhonghuai; He, Ningjia

    2016-01-01

    Genes from the plant specific Lateral Organ Boundaries Domain (LBD) family encode transcriptional regulators that have a variety of functions in various physiological and developmental processes. In the present study, 31 LBD genes were identified in the mulberry genome. The genome features of all MnLBD genes and phylogenetic studies with Arabidopsis LBD protein sequences, accompanied by the expression analysis of each of the Morus LBD genes provide insights into the functional prediction of mulberry LBDs. The genome-wide surveys of the current mulberry genome have resulted in the identification of catalogs of MnLBD genes that may function in the development of leaf, root, and secondary metabolism in Morus sp. PMID:27014591

  12. Antidiabetic and Antioxidant Effects and Phytochemicals of Mulberry Fruit (Morus alba L.) Polyphenol Enhanced Extract

    PubMed Central

    Wang, Yihai; Xiang, Limin; Wang, Chunhua; Tang, Chao; He, Xiangjiu

    2013-01-01

    The antidiabetic and antioxidant activities of the ethyl acetate-soluble extract (MFE) of mulberry fruit (Morus alba L.) were investigated. In vitro, MFE showed potent α-glucosidase inhibitory activity and radical-scavenging activities against DPPH and superoxide anion radicals. In vivo, MFE could significantly decrease fasting blood glucose (FBG) and glycosylated serum protein (GSP), and increase antioxidant enzymatic activities (SOD, CAT, GSH-Px) in streptozotocin (STZ)-induced diabetic mice. Bioactivity-guided fractionation of the MFE led to the isolation of 25 phenolic compounds, and their structures were identified on the basis of MS and NMR data. All the 25 compounds were isolated from mulberry fruit for the first time. Also, the α-glucosidase inhibitory activity and antioxidant activity of the phenolics were evaluated. Potent α-glucosidase inhibitory and radical-scavenging activities of these phenolics suggested that they may be partially responsible for the antidiabetic and antioxidant activities of mulberry fruit. PMID:23936259

  13. Sugarcane and mulberry silage supplementation of sheep during the peripartum period.

    PubMed

    David, Caroline Marçal Gomes; Costa, Ricardo Lopes Dias da; Parren, Guadalupe Aparecida Espicaski; Rua, Miguel Alejandro Silva; Nordi, Ellen Carolina Pereira; Okamoto, Fumiko; Paz, Cláudia Cristina Paro

    2015-04-01

    The aim of this study was to evaluate the effects of supplementation with silage made from sugarcane alone or mulberry and sugarcane on the gastrointestinal nematodes and hematological parameters in peripartum Santa Inês ewes. Three groups, each of 15 ewes, were supplemented with either 100 % sugarcane silage or sugarcane silage and mulberry at ratios of either 75:25 or 50:50. The variables analyzed were weight, body condition, packed cell volume, total plasma protein, hemoglobin, total leukocytes, eosinophils, and number of parasite eggs per gram of feces. All variables were measured individually every 14 days, making ten samples per ewe. Diets with sugarcane added to different proportions of mulberry branches did not influence the blood and parasitological parameters of Santa Inês sheep naturally infected in the peripartum period. PMID:25761641

  14. Reduction of post-prandial hyperglycemia by mulberry tea in type-2 diabetes patients

    PubMed Central

    Banu, Shaheena; Jabir, Nasimudeen R.; Manjunath, Nanjappa C.; Khan, Mohd Shahnawaz; Ashraf, Ghulam Md; Kamal, Mohammad Amjad; Tabrez, Shams

    2014-01-01

    Aim The dietary contents have a very important role in the management of metabolic syndrome along with type 2 diabetes mellitus (T2DM). Indian diet contains a large amount of carbohydrates that set off unpredictable blood sugar fluctuations and leads to increased risk of diabetic complications. The aim of the present study was to identify the effect of mulberry tea in the reduction of abnormally high postprandial blood glucose (PPG) levels in T2DM patients. Methods The study design was follow-up T2DM, 20 diabetic patients were given plain tea (control) and 28 diabetic patients were given mulberry tea (test subject) to measure the effect of mulberry tea on fasting blood glucose and PPG levels. Fasting blood glucose samples were collected after a standard breakfast. The PPG levels were recorded after the consumption of 70 ml tea along with 1 teaspoon of sugar after 90 min in all 48 patients. Results Fasting blood glucose levels in control and test group samples were found to be 178.55 ± 35.61 and 153.50 ± 48.10, respectively. After the consumption of plain tea and mulberry tea, the PPG values were recorded as 287.20 ± 56.37 and 210.21 ± 58.73, respectively. A highly significant (p < 0.001) change in the PPG level was observed in response to mulberry tea in all the test patients compared with control. Moreover, the effect size was also found to be very large (1.31). Conclusion Mulberry tea suppresses postprandial rise of blood glucose levels after 90 min of its consumption. PMID:25561880

  15. Results of deep-well injection testing at Mulberry, Florida

    USGS Publications Warehouse

    Hickey, John J.; Wilson, W.E.

    1982-01-01

    At the Kaiser Aluminum and Chemical Corporation plant, Mulberry, Fla., high-chloride, acidic liquid wastes are injected into a dolomite section at depths below about 4,000 feet below land surface. In 1975, a satellite monitor well was drilled 2,291 feet from the injection well and a series of three injection tests were performed. Duration of the tests ranged from 240 to 10,020 minutes and injection rates ranged from 110 to 230 gallons per minute. Based on an evaluation of factors that affect hydraulic response, water-level data suitable for interpretation of hydraulic characteristics of the injection zone were identified to occur from 200 to 1,000 minutes during the 10,020-minute test. Transmissivity of the injection zone was computed to be within the range from 700 to 1,000 feet squared per day and storage coefficient of the injection zone was computed to be within the range from .00001 to .00005. The confining bed accepting most of the leakage appears to be the underlying bed. Also, it appears that the overlying beds are probably relatively impermeable and significantly retard the vertical movement of neutralized waste effluent. (USGS)

  16. A novel sucrose hydrolase from the bombycoid silkworms Bombyx mori, Trilocha varians, and Samia cynthia ricini with a substrate specificity for sucrose.

    PubMed

    Wang, Huabing; Kiuchi, Takashi; Katsuma, Susumu; Shimada, Toru

    2015-06-01

    Although membrane-associated sucrase activity has been detected in the midgut of various lepidopteran species, it has not yet been identified and characterized at the molecular level. In the present study, we identified a novel sucrose hydrolase (SUH) gene from the following three bombycoid silkworms: Bombyx mori, Trilocha varians, and Samia cynthia ricini and named them BmSuh, TvSuh, and ScSuh, respectively. The EST dataset showed that BmSuh is one of the major glycoside hydrolase genes in the larval midgut of B. mori. These genes were almost exclusively expressed in the larval midgut in all three species, mainly at the feeding stage. SUHs are classified into the glycoside hydrolase family 13 and show significant homology to insect maltases. Enzymatic assays revealed that recombinant SUHs were distinct from conventional maltases and exhibited substrate specificity for sucrose. The recombinant BmSUH was less sensitive to sugar-mimic alkaloids than TvSUH and ScSUH, which may explain the reason why the sucrase activity in the B. mori midgut was less affected by the sugar-mimic alkaloids derived from mulberry. PMID:25937576

  17. 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

  18. Mutation of a Cuticle Protein Gene, BmCPG10, Is Responsible for Silkworm Non-Moulting in the 2nd Instar Mutant

    PubMed Central

    Zhao, Qiaoling; Kang, Lequn; Xia, Dingguo; Qiu, Zhiyong; Tang, Shunming; Li, Muwang; Shen, Xingjia; Zhang, Guozheng

    2016-01-01

    In the silkworm, metamorphosis and moulting are regulated by ecdysone hormone and juvenile hormone. The subject in the present study is a silkworm mutant that does not moult in the 2nd instar (nm2). Genetic analysis indicated that the nm2 mutation is controlled by a recessive gene and is homozygous lethal. Based on positional cloning, nm2 was located in a region approximately 275 kb on the 5th linkage group by eleven SSR polymorphism markers. In this specific range, according to the transcriptional expression of thirteen genes and cloning, the relative expression level of the BmCPG10 gene that encodes a cuticle protein was lower than the expression level of the wild-type gene. Moreover, this gene’s structure differs from that of the wild-type gene: there is a deletion of 217 bp in its open reading frame, which resulted in a change in the protein it encoded. The BmCPG10 mRNA was detectable throughout silkworm development from the egg to the moth. This mRNA was low in the pre-moulting and moulting stages of each instar but was high in the gluttonous stage and in newly exuviated larvae. The BmCPG10 mRNA showed high expression levels in the epidermis, head and trachea, while the expression levels were low in the midgut, Malpighian tubule, prothoracic gland, haemolymph and ventral nerve cord. The ecdysone titre was determined by ELISA, and the results demonstrated that the ecdysone titre of nm2 larvae was lower than that of the wild-type larvae. The nm2 mutant could be rescued by feeding 20-hydroxyecdysone, cholesterol and 7—dehydrocholesterol (7dC), but the rescued nm2 only developed to the 4th instar and subsequently died. The moulting time of silkworms could be delayed by BmCPG10 RNAi. Thus, we speculated that the mutation of BmCPG10 was responsible for the silkworm mutant that did not moult in the 2nd instar. PMID:27096617

  19. 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...

  20. 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

  1. Comparative study of aromatic compounds in fruit wines from raspberry, strawberry, and mulberry in central Shaanxi area

    PubMed Central

    Feng, Yiming; Liu, Min; Ouyang, Yanan; Zhao, Xianfang; Ju, Yanlun; Fang, Yulin

    2015-01-01

    Background Although grape wines have firmly dominated the production and consumption markets of fruit wines, raspberry, strawberry, and mulberry have been utilized to make wines because of their joyful aroma and high contents of polyphenolic phytochemicals and essential fatty acids. However, little is known about aromatic compounds of the wines produced from these three fruits. Methods The aromatic composition of fruit wines produced from raspberry, strawberry, mulberry, and red grape was analyzed by GC-MS. Odor activity values (OAVs) and relative odor contributions (ROCs) were used to estimate the sensory contribution of the aromatic compounds to the overall flavor of the wines. Results In strawberry, raspberry, and mulberry wines, 27, 30, and 31 odorants were detected, respectively. Alcohols formed the most abundant group, followed by esters and acids. The grape wine contained a wider variety (16 types) of alcohols, and 4-methyl-2-pentanol and 2,3-butanediol were not present in the three fruit wines. The quantity of esters in raspberry (1.54%) and mulberry wines (2.08%) were higher than those of strawberry wine (0.78%), and mulberry wine contained more types of esters. There were no significant differences of acids between the three fruit wines and the control wine. In addition, 2-heptanone, 2-octanone, 2-nonanone, and 2-undecanone were unique to raspberry wine, and nonanal was present only in mulberry wine. The indistinguishable aroma of the three fruit wines was attributed to the dominance of fruity and floral odor components derived from ethyl esters of fatty acids and their contributions to the global aroma of the three fruit wines. Conclusion The present study demonstrated that there were significant differences in the volatile components of fruit wines made from raspberry, strawberry, and mulberry. The aroma compounds were more abundant in the raspberry and mulberry wines than in the strawberry wine, but the quality of strawberry wine was superior to raspberry and mulberry wines. PMID:26617387

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

    PubMed

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

    2016-01-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2015-10-01

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

  5. Specific detection and identification of mulberry-infecting strains of Xylella fastidiosa by polymerase chain reaction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    X. fastidiosa causes bacterial leaf scorch in many landscape trees including elm, oak, sycamore and mulberry, but methods for specific identification of a particular tree host species-limited strain or differentiation of tree-specific strains are lacking. It is also unknown whether a particular land...

  6. Effect of fortification of Mulberry leaves with homeopathic drug Nux vomica on Bombyx mori. L.

    PubMed

    Hiware, C J

    2006-07-01

    Silk worm (Bombyx mori L.) larvae were fed on Mulberry leaves treated with Nux vomica mother tincture. The impact on larval, cocoon, shell and pupal weight, silk ratio, average filament length and denier, and number of breakages during reeling were investigated. The results were positive in all parameters under study except cocoon weight, pupal weight, and the average denier of the filament. PMID:16815518

  7. A holistic picture of Austronesian migrations revealed by phylogeography of Pacific paper mulberry.

    PubMed

    Chang, Chi-Shan; Liu, Hsiao-Lei; Moncada, Ximena; Seelenfreund, Andrea; Seelenfreund, Daniela; Chung, Kuo-Fang

    2015-11-01

    The peopling of Remote Oceanic islands by Austronesian speakers is a fascinating and yet contentious part of human prehistory. Linguistic, archaeological, and genetic studies have shown the complex nature of the process in which different components that helped to shape Lapita culture in Near Oceania each have their own unique history. Important evidence points to Taiwan as an Austronesian ancestral homeland with a more distant origin in South China, whereas alternative models favor South China to North Vietnam or a Southeast Asian origin. We test these propositions by studying phylogeography of paper mulberry, a common East Asian tree species introduced and clonally propagated since prehistoric times across the Pacific for making barkcloth, a practical and symbolic component of Austronesian cultures. Using the hypervariable chloroplast ndhF-rpl32 sequences of 604 samples collected from East Asia, Southeast Asia, and Oceanic islands (including 19 historical herbarium specimens from Near and Remote Oceania), 48 haplotypes are detected and haplotype cp-17 is predominant in both Near and Remote Oceania. Because cp-17 has an unambiguous Taiwanese origin and cp-17-carrying Oceanic paper mulberries are clonally propagated, our data concur with expectations of Taiwan as the Austronesian homeland, providing circumstantial support for the "out of Taiwan" hypothesis. Our data also provide insights into the dispersal of paper mulberry from South China "into North Taiwan," the "out of South China-Indochina" expansion to New Guinea, and the geographic origins of post-European introductions of paper mulberry into Oceania. PMID:26438853

  8. Improved 1-Deoxynojirimycin (DNJ) production in mulberry leaves fermented by microorganism

    PubMed Central

    Jiang, Yun-Gang; Wang, Chu-Yan; Jin, Chao; Jia, Jun-Qiang; Guo, Xijie; Zhang, Guo-Zheng; Gui, Zhong-Zheng

    2014-01-01

    DNJ, an inhibitor of α-glucosidase, is used to suppress the elevation of postprandial hyperglycemia. In this study, we focus on screening an appropriate microorganism for performing fermentation to improve DNJ content in mulberry leaf. Results showed that Ganoderma lucidum was selected from 8 species and shown to be the most effective in improvement of DNJ production from mulberry leaves through fermentation. Based on single factor and three factor influence level tests by following the Plackett-Burman design, the optimum extraction yield was analyzed by response surface methodology (RSM). The extracted DNJ was determined by reverse-phase high performance liquid chromatograph equipped with fluorescence detector (HPLC-FD). The results of RSM showed that the optimal condition for mulberry fermentation was defined as pH 6.97, potassium nitrate content 0.81% and inoculums volume 2 mL. The extraction efficiency reached to 0.548% in maximum which is 2.74 fold of those in mulberry leaf. PMID:25242964

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

    NASA Astrophysics Data System (ADS)

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

    2010-09-01

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

  10. 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

  11. Use of Sleeve Nets to Improve Survival of the Boisduval Silkworm, Anaphe panda, in the Kakamega Forest of Western Kenya

    PubMed Central

    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. Non-mulberry silk fibroin influence osteogenesis and osteoblast-macrophage cross talk on titanium based surface

    PubMed Central

    Naskar, Deboki; Nayak, Sunita; Dey, Tuli; Kundu, Subhas C.

    2014-01-01

    The titanium and its alloys are used as orthopedic dental implants due to their mechanical and bio-inert properties. The bare metal implants are not the ultimate answer for better osteogenesis and implant integration. Physical and chemical modifications are carried out to achieve the goal of improved adhesion and differentiation of the osteoblast. In this work, the silk fibroins from both mulberry and non-mulberry sources are used for surface modification. Silk fibroins are immobilized on titanium surface to facilitate the initial cell adhesion followed by improved cell spreading and better mineralization in order to achieve enhanced osseointegration. The immunological responses along with the effect of cytokines on osteoblast adhesion and function are investigated. The non-mulberry fibroin performs better in the context of the cell adherence and differentiation, which lead to better mineralization. The results indicate that the silk fibroin from non-mulberry source can be used for better osteogenesis on orthopedic implants. PMID:24752225

  13. Changes in chemical composition of a red wine aged in acacia, cherry, chestnut, mulberry, and oak wood barrels.

    PubMed

    De Rosso, Mirko; Panighel, Annarita; Dalla Vedova, Antonio; Stella, Laura; Flamini, Riccardo

    2009-03-11

    Aging in wooden barrels is a process used to stabilize the color and enrich the sensorial characteristics of wine. Many compounds are released from wood into the wine; oxygen permeation through the wood favors formation of new anthocyanin and tannin derivatives. Recently, polyphenols and volatile compounds released from acacia, chestnut, cherry, mulberry, and oak wood used in making barrels for spirits and wine aging were studied. Here, changes in volatile and polyphenolic compositions of a red wine aged for 9 months in acacia, cherry, chestnut, mulberry, and oak barrels are studied. Mulberry showed significant decreases of fruity-note ethyl esters and ethylguaiacol and a great cession of ethylphenol (horsey-odor defect). Cherry promoted the highest polyphenol oxidation, making it less suitable for long aging. LC/ESI-MS(n) showed the relevant presence of cis- and trans-piceatannol in mulberry-aged wine, a phytoalexin with antileukemia and antimelanoma activities. PMID:19196157

  14. Mulberry trees conserved soil and protected water quality in the riparian zone of the Three Gorges Reservoir, China.

    PubMed

    Liu, Yun; Willison, J H Martin; Wan, Pan; Xiong, Xing-Zheng; Ou, Yang; Huang, Xiao-Hui; Wu, Jingchun; Zhou, Hao; Xu, Qiao; Chen, Guohui; Xili, Yuanzi; Nie, Jiasheng

    2016-03-01

    China's Sloping Land Conversion Program (SLCP) was designed to restore perennial plant cover on sloping land in western China, in part to protect the Three Gorges Reservoir (TGR). In this study, we examined use of white mulberry (Morus alba L.) in the SLCP to protect water quality and conserve soil. We established nine runoff monitoring plots divided among three categories (vegetable farming, fallow control, and mulberry plantation) on a bank of the Liangtan River situated at the western margin of the TGR. The land had been used previously by farmers for growing vegetables. We found that soil loss and surface water runoff were lowest in the mulberry plots and highest in the vegetable plots. We used inductively coupled plasma atomic emission spectroscopy (ICP-AES) to assess the concentration of selected heavy metal pollution indicators (Zn, Hg, As, Ni, Pb, Cr, Cd, and Cu) in the monitoring plot soils at the beginning of the experiment in May 2009. The heavy metals were assessed again at the end of the experiment in October 2012, and we found that the concentrations of these pollutants had been reduced in all fallow and mulberry plots, and to the greatest extent in the mulberry plots. We found that levels of Hg, Pb, and Cu increased in the vegetable plots. For these reasons, we conclude that riparian mulberry plantations are useful for reducing rapid runoff of storm water, conserving soil, and sequestering heavy metal pollutants in the TGR region. PMID:26564183

  15. Effect of Cultivar, Temperature, and Environmental Conditions on the Dynamic Change of Melatonin in Mulberry Fruit Development and Wine Fermentation.

    PubMed

    Wang, Cheng; Yin, Li-Yuan; Shi, Xue-Ying; Xiao, Hua; Kang, Kun; Liu, Xing-Yan; Zhan, Ji-Cheng; Huang, Wei-Dong

    2016-04-01

    High levels of melatonin have been reported in various foods but not in mulberry or its wine. This study investigated the dynamic changes of melatonin levels during mulberry fruit development and ethanol fermentation of 2 different colored mulberry cultivars ("Hongguo2ˮ Morus nigra, black and "Baiyuwangˮ Morus alba, white) at 2 fermentation temperatures (16 and 25 °C). Our results showed that the melatonin level increased in the beginning of mulberry development but decreased in the end. The MnTDC gene expression level correlated with melatonin production, which implied that TDC may be the rate-limiting enzyme of the melatonin biosynthetic process in mulberries. During mulberry fermentation, the melatonin concentration increased rapidly in the beginning and then decreased gradually. Low temperature delayed the melatonin production during fermentation. A relatively high level of melatonin was found in "Hongguo2ˮ compared with "Baiyuwangˮ during fruit development and fermentation. The variation of melatonin correlated with the ethanol production rate, suggesting that melatonin may participate in physiological regulation of Saccharomyces cerevisiae during the fermentation stage. PMID:26953927

  16. Quantitative Changes of Polyphenolic Compounds in Mulberry (Morus alba L.) Leaves in Relation to Varieties, Harvest Period, and Heat Processing

    PubMed Central

    Lee, Won Jeong; Choi, Sang Won

    2012-01-01

    Six polyphenolic compounds, such as chlorogenic acid (CA), rutin (RT), isoquercitrin (IQT), quercetin-3-O-(6-O-malonyl)-β-D-glucoside (QMG), astragalin (AG), kaempferol-3-O-(6-O-malonyl)-β-D-glucoside (KMG), were isolated from mulberry leaves by a series of isolation procedures, such as Diaion HP-20, silica-gel, Sephadex LH-20, and ODS-A column chromatographies. The chemical structures of the phenolic compounds were identified by UV and NMR spectral analyses. Levels of polyphenols in mulberry leaves from six different mulberry cultivars ranged from 1,042.16 to 1,871.97 mg% per dry weight; Guksang cultivar showed the highest levels of polyphenols, whereas Gaeryangdaehwa contained the least polyphenol contents. Generally, levels of polyphenols in mulberry leaves decreased with increasing harvest time, except for Yoolmok, but increased with heat processing time, except QMG and KMG. These results suggest that the heat processed mulberry leaves of Guksang cultivar harvested in early May can be potentially useful sources for production of high quality mulberry leaf teas. PMID:24471097

  17. Proteomic Analysis of Larval Midgut from the Silkworm (Bombyx mori)

    PubMed Central

    Zhang, Sai; Xu, Yunmin; Fu, Qiang; Jia, Ling; Xiang, Zhonghuai; He, Ningjia

    2011-01-01

    The midgut is the major organ for food digestion, nutrient absorption and also a barrier for foreign substance. The 5th-instar larval stage of silkworm is very important for larval growth, development, and silk production. In the present study, we used 2-DE and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to analyze the midgut proteins from the 5th-instar larvae as well as the midgut proteins under starvation condition. A total of 96 proteins were identified in this study; and among them, 69 proteins were observed in midgut for the first time. We also found that the silkworm larval midgut responded to starvation by producing a 10 kDa heat shock protein and a diapause hormone precursor. PMID:21687556

  18. Transcriptome Analysis of Thermal Parthenogenesis of the Domesticated Silkworm

    PubMed Central

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

    2015-01-01

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

  19. Transgenic silkworms produce recombinant human type III procollagen in cocoons.

    PubMed

    Tomita, Masahiro; Munetsuna, Hiroto; Sato, Tsutomu; Adachi, Takahiro; Hino, Rika; Hayashi, Masahiro; Shimizu, Katsuhiko; Nakamura, Namiko; Tamura, Toshiki; Yoshizato, Katsutoshi

    2003-01-01

    We describe the generation of transgenic silkworms that produce cocoons containing recombinant human collagen. A fusion cDNA was constructed encoding a protein that incorporated a human type III procollagen mini-chain with C-propeptide deleted, a fibroin light chain (L-chain), and an enhanced green fluorescent protein (EGFP). This cDNA was ligated downstream of the fibroin L-chain promoter and inserted into a piggyBac vector. Silkworm eggs were injected with the vectors, producing worms displaying EGFP fluorescence in their silk glands. The cocoons emitted EGFP fluorescence, indicating that the promoter and fibroin L-chain cDNAs directed the synthesized products to be secreted into cocoons. The presence of fusion proteins in cocoons was demonstrated by immunoblotting, collagenase-sensitivity tests, and amino acid sequencing. The fusion proteins from cocoons were purified to a single electrophoretic band. This study demonstrates the viability of transgenic silkworms as a tool for producing useful proteins in bulk. PMID:12483223

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

    PubMed

    Sekimizu, N; Paudel, A; Hamamoto, H

    2012-08-01

    Sacrificing model animals is required for developing effective drugs before being used in human beings. In Japan today, at least 4,210,000 mice and other mammals are sacrificed to a total of 6,140,000 per year for the purpose of medical studies. All the animals treated in Japan, including test animals, are managed under control of "Act on Welfare and Management of Animals". Under the principle of this Act, no person shall kill, injure, or inflict cruelty on animals without due cause. "Animal" addressed in the Act can be defined as a "vertebrate animal". If we can make use of invertebrate animals in testing instead of vertebrate ones, that would be a remarkable solution for the issue of animal welfare. Furthermore, there are numerous advantages of using invertebrate animal models: less space and small equipment are enough for taking care of a large number of animals and thus are cost-effective, they can be easily handled, and many biological processes and genes are conserved between mammals and invertebrates. Today, many invertebrates have been used as animal models, but silkworms have many beneficial traits compared to mammals as well as other insects. In a Genome Pharmaceutical Institute's study, we were able to achieve a lot making use of silkworms as model animals. We would like to suggest that pharmaceutical companies and institutes consider the use of the silkworm as a model animal which is efficacious both for financial value by cost cutting and ethical aspects in animals' welfare. PMID:23006994

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

    PubMed

    Sumriddetchkajorn, Sarun; Kamtongdee, Chakkrit

    2012-02-01

    This paper proposes and experimentally demonstrates for what is believed to be the first time a highly sought-after optical structure for highly-accurate identification of the silkworm pupa gender. The key idea is to exploit a long wavelength optical beam in the red or near infrared spectrum that can effectively and safely penetrate the body of a silkworm pupa. Later on, simple image processing operations via image thresholding, blob filtering, and image inversion processes are applied in order to eliminate the unwanted image noises and at the same time highlight the gender gland. Experimental proof of concept using three 636 nm wavelength light emitting diodes, a two-dimensional web camera, an 8 bit microcontroller board, and a notebook computer shows a very high 95.6% total accuracy in identifying the gender of 45 silkworm pupae with a measured fast identification time of 96.6 ms. Other key features include low cost, low component counts, and ease of implementation and control. PMID:22307109

  2. 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

  3. 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.

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

    PubMed

    Nishida, Satoshi; Ono, Yasuo; Sekimizu, Kazuhisa

    2016-01-01

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

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

    NASA Astrophysics Data System (ADS)

    Kamtongdee, Chakkrit; Sumriddetchkajorn, Sarun; Chanhorm, Sataporn

    2015-07-01

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

  6. Proteomic analysis of nucleopolyhedrovirus infection resistance in the silkworm, Bombyx mori (Lepidoptera: Bombycidae).

    PubMed

    Liu, Xiaoyong; Yao, Qin; Wang, Yong; Chen, Keping

    2010-09-01

    Silkworm hemolymph is an important defense tissue to resist bacteria and virus infections. To study the response of silkworm hemolymph in the resistance of Bombyx mori L. nucleopolyhedrovirus (BmNPV), we constructed a near-isogenic silkworm line with BmNPV resistance using highly resistant and highly susceptible parental strains. In this paper, two-dimensional gel electrophoresis (2-DE) and Matrix-Assisted Laser Desorption/Ionization (MALDI)-mass spectrometry were employed to investigate the differences of protein patterns in the hemolymph of the highly resistant, highly susceptible and near-isogenic silkworm strains after BmNPV was administrated to the larvae. A comparison between the proteomes of these three silkworm strains led us to identify two differentially expressed proteins, beta-N-acetylglucosaminidase 2 and aminoacylase. The expression levels of these proteins were higher in the BmNPV resistant strains. PMID:20471391

  7. Wild Animals.

    ERIC Educational Resources Information Center

    Web Feet K-8, 2000

    2000-01-01

    This annotated subject guide to Web sites and other resources focuses on wild animals. Includes Web sites, CD-ROMs and software, videos, books, audios, magazines, and professional resources, as well as a class activity. (LRW)

  8. A holistic picture of Austronesian migrations revealed by phylogeography of Pacific paper mulberry

    PubMed Central

    Chang, Chi-Shan; Liu, Hsiao-Lei; Moncada, Ximena; Seelenfreund, Andrea; Seelenfreund, Daniela; Chung, Kuo-Fang

    2015-01-01

    The peopling of Remote Oceanic islands by Austronesian speakers is a fascinating and yet contentious part of human prehistory. Linguistic, archaeological, and genetic studies have shown the complex nature of the process in which different components that helped to shape Lapita culture in Near Oceania each have their own unique history. Important evidence points to Taiwan as an Austronesian ancestral homeland with a more distant origin in South China, whereas alternative models favor South China to North Vietnam or a Southeast Asian origin. We test these propositions by studying phylogeography of paper mulberry, a common East Asian tree species introduced and clonally propagated since prehistoric times across the Pacific for making barkcloth, a practical and symbolic component of Austronesian cultures. Using the hypervariable chloroplast ndhF-rpl32 sequences of 604 samples collected from East Asia, Southeast Asia, and Oceanic islands (including 19 historical herbarium specimens from Near and Remote Oceania), 48 haplotypes are detected and haplotype cp-17 is predominant in both Near and Remote Oceania. Because cp-17 has an unambiguous Taiwanese origin and cp-17–carrying Oceanic paper mulberries are clonally propagated, our data concur with expectations of Taiwan as the Austronesian homeland, providing circumstantial support for the “out of Taiwan” hypothesis. Our data also provide insights into the dispersal of paper mulberry from South China “into North Taiwan,” the “out of South China–Indochina” expansion to New Guinea, and the geographic origins of post-European introductions of paper mulberry into Oceania. PMID:26438853

  9. Quantification of Changes in Mulberry Silk Fabrics due to Different Laundering: Using WAXS Technique

    NASA Astrophysics Data System (ADS)

    Parameswara, P.; Nivedita, S.; Somashekar, R.

    2011-07-01

    Loom finished mulberry silk fabrics (Taffeta) were machine laundered and hand laundered several times. X-ray diffractograms of pure and laundered fabrics were used to calculate microstructural parameters like average crystallite size (D) and lattice strain (Vegr) employing Williamson-Hall plot. Microstructural parameters were compared with measured mechanical properties like breaking load, tenacity, and elongation of warp yarns unraveled from fabrics. Surface morphology and texture of silk fabrics changed upon washing is evident from SEM images.

  10. Anti-inflammatory and anti-cancer activity of mulberry (Morus alba L.) root bark

    PubMed Central

    2014-01-01

    Background Root bark of mulberry (Morus alba L.) has been used in herbal medicine as anti-phlogistic, liver protective, kidney protective, hypotensive, diuretic, anti-cough and analgesic agent. However, the anti-cancer activity and the potential anti-cancer mechanisms of mulberry root bark have not been elucidated. We performed in vitro study to investigate whether mulberry root bark extract (MRBE) shows anti-inflammatory and anti-cancer activity. Methods In anti-inflammatory activity, NO was measured using the griess method. iNOS and proteins regulating NF-κB and ERK1/2 signaling were analyzed by Western blot. In anti-cancer activity, cell growth was measured by MTT assay. Cleaved PARP, ATF3 and cyclin D1 were analyzed by Western blot. Results In anti-inflammatory effect, MRBE blocked NO production via suppressing iNOS over-expression in LPS-stimulated RAW264.7 cells. In addition, MRBE inhibited NF-κB activation through p65 nuclear translocation via blocking IκB-α degradation and ERK1/2 activation via its hyper-phosphorylation. In anti-cancer activity, MRBE deos-dependently induced cell growth arrest and apoptosis in human colorectal cancer cells, SW480. MRBE treatment to SW480 cells activated ATF3 expression and down-regulated cyclin D1 level. We also observed that MRBE-induced ATF3 expression was dependent on ROS and GSK3β. Moreover, MRBE-induced cyclin D1 down-regulation was mediated from cyclin D1 proteasomal degradation, which was dependent on ROS. Conclusions These findings suggest that mulberry root bark exerts anti-inflammatory and anti-cancer activity. PMID:24962785

  11. Food-grade mulberry powder enriched with 1-deoxynojirimycin suppresses the elevation of postprandial blood glucose in humans.

    PubMed

    Kimura, Toshiyuki; Nakagawa, Kiyotaka; Kubota, Hiroyuki; Kojima, Yoshihiro; Goto, Yuko; Yamagishi, Kenji; Oita, Shigeru; Oikawa, Shinichi; Miyazawa, Teruo

    2007-07-11

    Mulberry 1-deoxynojirimycin (DNJ), a potent glucosidase inhibitor, has been hypothesized to be beneficial for the suppression of abnormally high blood glucose levels and thereby prevention of diabetes mellitus. However, DNJ contents in commercial mulberry products were as low as about 0.1% (100 mg/100 g of dry product), implying that the bioavailability of DNJ might not be expected. We carried out studies in two directions: (1) production of food-grade mulberry powder containing a maximally high DNJ content; (2) determination of the optimal dose of the DNJ-enriched powder for the suppression of the postprandial blood glucose through clinical trials. The following method was used: (1) DNJ concentrations in mulberry leaves from different cultivars, harvest seasons, and leaf locations were determined using hydrophilic interaction chromatography with evaporative light scattering detection. (2) Healthy volunteers received 0, 0.4, 0.8, and 1.2 g of DNJ-enriched powder (corresponding to 0, 6, 12, and 18 mg of DNJ, respectively), followed by 50 g of sucrose. Before and 30-180 min after the DNJ/sucrose administration, plasma glucose and insulin were determined. The following results were obtained: (1) Young mulberry leaves taken from the top part of the branches in summer contained the highest amount of DNJ. After optimization of the harvesting and drying processes for young mulberry leaves (Morus alba L. var. Shin ichinose), DNJ-enriched powder (1.5%) was produced. (2) A human study indicated that the single oral administration of 0.8 and 1.2 g of DNJ-enriched powder significantly suppressed the elevation of postprandial blood glucose and secretion of insulin, revealing the physiological impact of mulberry DNJ (effective dose and efficacy in humans). This study suggests that the newly developed DNJ-enriched powder can be used as a dietary supplement for preventing diabetes mellitus. PMID:17555327

  12. Optimization for ultrasound extraction of polysaccharides from mulberry fruits with antioxidant and hyperglycemic activity in vitro.

    PubMed

    Chen, Chun; You, Li-Jun; Abbasi, Arshad Mehmood; Fu, Xiong; Liu, Rui Hai

    2015-10-01

    Single-factor experiment and Box-Behnken design (BBD) were applied to optimize the ultrasound-assisted extraction of mulberry fruits polysaccharides (MFP). Under optimum conditions: ratio of water to raw material 40.25, extraction temperature 69°C, ultrasonic power 190W and extraction time 75 min, the MFP yield was 3.13% (±0.07%), in accordance to the predicted value of 3.04%. The mulberry fruits polysaccharides fractions was obtained by deproteinization (MFP-1), followed by decolorization and deionization (MFP-2). Carbohydrate content in MFP, MFP-1 and MFP-2 was 58.61% (±1.47%), 69.98% (±0.91%), 81.18% (±1.29%), as well as proteins was estimated 16.50% (±0.86%), 1.57% (±0.63%), 1.02% (±0.18%), respectively. The FT-IR indicated that MFP, MFP-1 and MFP-2 were acidic polysaccharides. The MFP-1 exhibited the strongest antioxidant activity, while MFP-2 showed the strongest hyperglycemic activity in vitro. This may be caused by their different compositions and physical properties in the different mulberry fruit polysaccharides fractions. PMID:26076608

  13. Bioactive compounds and sensory quality of black and white mulberries grown in Spain.

    PubMed

    Calín-Sánchez, Angel; Martínez-Nicolás, Juan José; Munera-Picazo, Sandra; Carbonell-Barrachina, Angel A; Legua, Pilar; Hernández, Francisca

    2013-12-01

    The objective of this study was to evaluate and compare, for the first time, white and black mulberry species in terms of main phytochemical, volatile composition and sensory profile characteristics in eight Spanish clones. The results showed that black and white mulberry species displayed significant different characteristics. PLS analysis has allowed grouping of the clones into four groups (i) MA1, MA2 and MN2, (ii) MN3 and MN4, (iii) MA3 and MA4, and (iv) MN1. Experimental results proved that Spanish mulberries have potential for fresh consumption due to their high antioxidant capacity (10.7-86.1 mg Trolox 100 g(-1)), polyphenol (76.7-180 mg gallic acid 100 g(-1)) and ellagic acid content (8.7-15.5 mg 100 g(-1)) as well as considerable amount of volatile compounds (35) with desirable attributes, which were scored high by a trained panel. Cultivars from the Morus nigra species seem to provide fruits with higher content of bioactive compounds and better aptitude for fresh consumption than Morus alba. Differences among the species should be attributed to genetics because they were cultivated under same conditions. PMID:23979495

  14. Persistent stimulation of photosynthesis in short rotation coppice mulberry under elevated CO2 atmosphere.

    PubMed

    Madhana Sekhar, Kalva; Rachapudi, Venkata Sreeharsha; Mudalkar, Shalini; Reddy, Attipalli Ramachandra

    2014-08-01

    Current study was undertaken to elucidate the responses of short rotation coppice (SRC) mulberry under elevated CO2 atmosphere (550?molmol(-1)). Throughout the experimental period, elevated CO2 grown mulberry plants showed significant increase in light saturated photosynthetic rates (A') by increasing intercellular CO2 concentrations (Ci) despite reduced stomatal conductance (gs). Reduced gs was linked to decrease in transpiration (E) resulting in improved water use efficiency (WUE). There was a significant increase in carboxylation efficiency (CE) of Rubisco, apparent quantum efficiency (AQE), light and CO2 saturated photosynthetic rates (AMAX), photosynthetic nitrogen use efficiency (PNUE), chlorophyll a fluorescence characteristics (FV/FM and PIABS), starch and other carbohydrates in high CO2 grown plants which clearly demonstrate no photosynthetic acclimation in turn resulted marked increase in above and below ground biomass. Our results strongly suggest that short rotation forestry (<1year) with mulberry plantations should be effective to mitigate raising CO2 levels as well as for the production of renewable bio-energy. PMID:24938741

  15. Phenolic acids profiling and antioxidant potential of mulberry (Morus laevigata W., Morus nigra L., Morus alba L.) leaves and fruits grown in Pakistan

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mulberry trees are distributed throughout Pakistan. Besides the use of mulberry in forage and food for animals, it is also used as herbal medicine. The ojbective of this study was to determine phenolic acids profile, sugar content, and the antioxidant activity of the leaves and fruits of three mulb...

  16. The Quality Changes of Postharvest Mulberry Fruit Treated by Chitosan-g-Caffeic Acid during Cold Storage.

    PubMed

    Yang, Caifeng; Han, Beibei; Zheng, Yu; Liu, Lili; Li, Changlong; Sheng, Sheng; Zhang, Jian; Wang, Jun; Wu, Fuan

    2016-04-01

    This study aimed to characterize the effects of chitosan-g-caffeic acid (CTS-g-CA) on improving the quality and extending the shelf life of postharvest mulberry fruit during storage at 4 °C for 18 d. CTS-g-CA was enzymatically synthesized using laccase from Pleurotus ostreatus as a catalyst. The synergistic effects of CTS-g-CA treatment on mulberry fruit were evaluated using a co-toxicity factor (cf). The results showed that the rotting rate of CTS-g-CA-treated fruit was 37.67% (compared with that of the control at 97.67%) on day 18. The weight loss and malondialdehyde (MDA) contents of the CTS-g-CA-treated mulberry fruit were significantly lower (P < 0.05) than those of the control, CA, CTS, and CA+CTS treatments. Moreover, the DPPH and ABTS radical scavenging activities of the CTS-g-CA treatment were both higher than those of the control. Furthermore, the CTS-g-CA treatment also maintained higher levels of main active substances, such as anthocyanins, ascorbic acid, polyphenols and flavones, in mulberry fruit than the other treatments. Therefore, CTS-g-CA could be used to improve the quality and extend the shelf life of postharvest mulberry fruit during cold storage. PMID:26992122

  17. Identification and Determination of the Polyhydroxylated Alkaloids Compounds with α-Glucosidase Inhibitor Activity in Mulberry Leaves of Different Origins.

    PubMed

    Ji, Tao; Li, Jun; Su, Shu-Lan; Zhu, Zhen-Hua; Guo, Sheng; Qian, Da-Wei; Duan, Jin-Ao

    2016-01-01

    Mulberry leaves have commonly been utilized in China as a herbal medicine for the treatment of diabetes for thousands of years. To evaluate the quality, an ultra-high performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS) method was developed for identification of polyhydroxylated alkaloids with α-glucosidase inhibitor activity in mulberry leaf. As a result, five alkaloid compounds were identified or tentatively characterized. Among them, the compound 1-deoxynojirimycin (DNJ) was selected as the most typical and active chemical marker and quantified using an improved high performance liquid chromatography (HPLC) normal phase coupled with evaporative light scattering detector (ELSD) method. The developed method was fully validated in terms of linearity, sensitivity, precision and repeatability, as well as recovery, and subsequently applied to evaluate twenty-nine batches of mulberry leaves from different collections. From the analytical data it was discovered that the average content of DNJ is 1.53 mg/g, while the total contents of DNJ in the 29 mulberry leaf sample ranged from 0.20 to 3.88 mg/g, which suggested remarkable differences, although it reached the highest levels in early August. These data may provide an important reference for the quality of mulberry leaves used as herbal medicine for the treatment of diabetes or as a material to obtain the DNJ of α-glucosidase inhibitor or as a functional food. PMID:26867190

  18. Hyperspectral Imaging Coupled with Random Frog and Calibration Models for Assessment of Total Soluble Solids in Mulberries

    PubMed Central

    Zhao, Yan-Ru; Yu, Ke-Qiang; He, Yong

    2015-01-01

    Chemometrics methods coupled with hyperspectral imaging technology in visible and near infrared (Vis/NIR) region (380–1030 nm) were introduced to assess total soluble solids (TSS) in mulberries. Hyperspectral images of 310 mulberries were acquired by hyperspectral reflectance imaging system (512 bands) and their corresponding TSS contents were measured by a Brix meter. Random frog (RF) method was used to select important wavelengths from the full wavelengths. TSS values in mulberry fruits were predicted by partial least squares regression (PLSR) and least-square support vector machine (LS-SVM) models based on full wavelengths and the selected important wavelengths. The optimal PLSR model with 23 important wavelengths was employed to visualise the spatial distribution of TSS in tested samples, and TSS concentrations in mulberries were revealed through the TSS spatial distribution. The results declared that hyperspectral imaging is promising for determining the spatial distribution of TSS content in mulberry fruits, which provides a reference for detecting the internal quality of fruits. PMID:26451273

  19. Prospects for cultivating white mulberry (Morus alba) in the drawdown zone of the Three Gorges Reservoir, China.

    PubMed

    Liu, Yun; Willison, J H Martin

    2013-10-01

    Restoration of vegetation is the most viable management approach for restoring ecological functions in the drawdown zone (hydro-fluctuation belt) of the Three Gorges Reservoir. The selection of plants for this purpose is therefore critically important. Most indigenous plants are not adapted, however, to the counter-seasonal fluctuation of water levels and rapid changes of up to 30 m in water depth that characterize the management of the reservoir. As a result, the reservoir drawdown zone tends to be vegetation deficient. Mulberry (Morus alba L.) has attracted attention as a suitable woody plant for restoring woody vegetation because of its strong adaptation to environmental stresses and the finding that it survives up to 7 m of flooding in parts of the drawdown zone. Comprehensive evaluation of research is therefore required in order to provide guidance for the rational use of mulberry in vegetation restoration strategies for the drawdown zone. Knowledge of the physiology of mulberry adaptation to stress is reviewed here, along with a detailed review of the ecology and agricultural benefits and limitations of mulberry in the context of the Three Gorges Reservoir. It is proposed that a cultivation model for mulberry plants based on ecological principles should be adopted for use within the drawdown zone and that a wider range of biophysical and socio-economic research to develop this model further should be conducted in the future. PMID:23757029

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

    NASA Astrophysics Data System (ADS)

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

    2009-06-01

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

  1. 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

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

    PubMed Central

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

    2013-01-01

    The silkworm, Bombyx mori, is one of the major insect model organisms, and its draft and fine genome sequences became available in 2004 and 2008, respectively. Transposable elements (TEs) constitute ∼40% of the silkworm genome. To better understand the roles of TEs in organization, structure and evolution of the silkworm genome, we used a combination of de novo, structure-based and homology-based approaches for identification of the silkworm TEs and identified 1308 silkworm TE families. These TE families and their classification information were organized into a comprehensive and easy-to-use web-based database, BmTEdb. Users are entitled to browse, search and download the sequences in the database. Sequence analyses such as BLAST, HMMER and EMBOSS GetORF were also provided in BmTEdb. This database will facilitate studies for the silkworm genomics, the TE functions in the silkworm and the comparative analysis of the insect TEs. Database URL: http://gene.cqu.edu.cn/BmTEdb/. PMID:23886610

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

    PubMed Central

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

    2001-01-01

    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

  4. 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.

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

    SciTech Connect

    Chen Fujia; Porter, David; Vollrath, Fritz

    2010-10-15

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

  6. Microarray-based gene expression profiles in multiple tissues of the domesticated silkworm, Bombyx mori.

    PubMed

    Xia, Qingyou; Cheng, Daojun; Duan, Jun; Wang, Genhong; Cheng, Tingcai; Zha, Xingfu; Liu, Chun; Zhao, Ping; Dai, Fangyin; Zhang, Ze; He, Ningjia; Zhang, Liang; Xiang, Zhonghuai

    2007-01-01

    We designed and constructed a genome-wide microarray with 22,987 70-mer oligonucleotides covering the presently known and predicted genes in the silkworm genome, and surveyed the gene expression in multiple silkworm tissues on day 3 of the fifth instar. Clusters of tissue-prevalent and tissue-specific genes and genes that are differentially expressed in different tissues were identified, and they reflect well major tissue-specific functions on the molecular level. The data presented in this study provide a new resource for annotating the silkworm genome. PMID:17683582

  7. Wild Marshmallows.

    ERIC Educational Resources Information Center

    Kallas, John N.

    1984-01-01

    Provides information for teaching a unit on wild plants, including resources to use, plants to learn, safety considerations, list of plants (with scientific name, edible parts, and uses), list of plants that might cause allergic reactions when eaten. Also describes the chickweed, bull thistle, and common mallow. (BC)

  8. Effect of Ensiled Mulberry Leaves and Sun-Dried Mulberry Fruit Pomace on Finishing Steer Growth Performance, Blood Biochemical Parameters, and Carcass Characteristics

    PubMed Central

    Zhou, Zhenming; Zhou, Bo; Ren, Liping; Meng, Qingxiang

    2014-01-01

    Fifty-one Simmental crossbred steers (357.0±16.5 kg) were used to compare a standard total mix ration (TMR) with variants on animal performance, ruminal fermentation, blood biochemical parameters, and carcass characteristics. Corn grain and cotton seed meal were partially replaced by ensiled mulberry leaves (EML) or sun-dried mulberry fruit pomace (SMFP). Experimental diets had similar amounts of crude protein (CP), acid detergent fiber (ADF), and metabolizable energy (ME). Animals were divided into three groups: control group (CONT), 8% EML group, and 6.3% SMFP group. Performance, including average daily weight gain (ADG), and dry matter intake (DMI), was measured. Blood and rumen samples were collected at the end of the experiment (16 weeks). There were no differences in final body weight (P = 0.743), ADG (P = 0.425), DMI (P = 0.642), or ADG/DMI (P = 0.236) between the groups. There were no differences (P = 0.2024) in rumen pH values; ammonia N was lower (P = 0.0076) in SMFP than in the EML and CONT groups. There were differences in the concentrations of total and individual volatile fatty acids, while no differences were determined in blood biochemical parameters (i.e., plasma glucose, urea concentrations, triglycerides, total protein, insulin, IgG, alanine transaminase, and aspartate aminotransferase, P ≥ 0.098). No differences were observed in carcass characteristics (P ≥ 0.513), tenderness (P = 0.844), adipose and lean color values (P ≥ 0.149), and chemical composition (P ≥ 0.400); however, intramuscular fat was lower in the EML and SMFP groups compared to the CONT animals (P = 0.034). In conclusion, diets supplemented with these two mulberry products in an isocaloric and isonitrogenous manner have similar effects to corn grain and cotton seed meals on steer performance, blood biochemical parameters and carcass characteristics, with the exception of ruminal VFA concentrations and lower intramuscular fat content. PMID:24427304

  9. The complete mitochondrial genome of the wild silkworm moth, Actias selene.

    PubMed

    Liu, Qiu-Ning; Zhu, Bao-Jian; Dai, Li-Shang; Wei, Guo-Qing; Liu, Chao-Liang

    2012-09-01

    The complete mitochondrial genome (mitogenome) of Actias selene (Lepidoptera: Saturniidae) was determined to be 15,236 bp, including 13 protein-coding genes (PCGs), two rRNA genes, 22 tRNA genes and a control region. The arrangement of 13 PCGs was similar to that of other sequenced lepidopterans. The AT skew of the mitogenome of A. selene was slightly negative, indicating a higher number of T compared to A nucleotides. The nucleotide composition of the mitogenome of A. selene was also biased toward A+T nucleotides (78.91%). All PCGs were initiated by ATN codons, except for the gene encoding cytochrome c oxidase subunit 1 (cox1), which may be initiated by the TTAG, as observed in other lepidopterans. Three genes, including cox1, cox2, and nad5, had incomplete stop codons consisting of just a T. With an exception for trnS1(AGN), all the other tRNA genes displayed a typical clover-leaf structure of mitochondrial tRNA. The A+T-rich region of the mitogenome of A. selene was 339 bp in length, and contains several features common to the Lepidopteras, including non-repetitive sequences, a conserved structure combining the motif ATAGA and an 18-bp poly-T stretch and a poly-A element upstream of trnM gene. Phylogenetic analysis showed that A. selene was close to Saturniidae. PMID:22688122

  10. The genetics and genomics of the silkworm, Bombyx mori.

    PubMed

    Goldsmith, Marian R; Shimada, Toru; Abe, Hiroaki

    2005-01-01

    We review progress in applying molecular genetic and genomic technologies to studies in the domesticated silkworm, Bombyx mori, highlighting its use as a model for Lepidoptera, and in sericulture and biotechnology. Dense molecular linkage maps are being integrated with classical linkage maps for positional cloning and marker-assisted selection. Classical mutations have been identified by a candidate gene approach. Cytogenetic and sequence analyses show that the W chromosome is composed largely of nested full-length long terminal repeat retrotransposons. Z-chromosome-linked sequences show a lack of dosage compensation. The downstream sex differentiation mechanism has been studied via the silkworm homolog of doublesex. Expressed sequence tagged databases have been used to discover Lepidoptera-specific genes, provide evidence for horizontal gene transfer, and construct microarrays. Physical maps using large-fragment bacterial artificial chromosome libraries have been constructed, and whole-genome shotgun sequencing is underway. Germline transformation and transient expression systems are well established and available for functional studies, high-level protein expression, and gene silencing via RNA interference. PMID:15355234

  11. Expression profile of cuticular genes of silkworm, Bombyx mori

    PubMed Central

    2010-01-01

    Background Insect cuticle plays essential roles in many physiological functions. During molting and metamorphosis tremendous changes occur in silkworm cuticle where multiple proteins exist and genes encoding them constitute about 1.5% of all Bombyx mori genes. Results In an effort to determine their expression profiles, a microarray-based investigation was carried out using mRNA collected from larvae to pupae. The results showed that a total of 6676 genes involved in various functions and physiological pathways were activated. The vast majority (93%) of cuticular protein genes were expressed in selected stages with varying expression patterns. There was no correlation between expression patterns and the presence of conserved motifs. Twenty-six RR genes distributed in chromosome 22 were co-expressed at the larval and wandering stages. The 2 kb upstream regions of these genes were further analyzed and three putative elements were identified. Conclusions Data from the present study provide, for the first time, a comprehensive expression profile of genes in silkworm epidermal tissues and evidence that putative elements exist to allow massive production of mRNAs from specific cuticular protein genes. PMID:20226095

  12. 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

  13. Mechanical behavior comparison of spider and silkworm silks using molecular dynamics at atomic scale.

    PubMed

    Lee, Myeongsang; Kwon, Junpyo; Na, Sungsoo

    2016-02-01

    Spider and silkworm silk proteins have received much attention owing to their inherent structural stability, biodegradability, and biocompatibility. These silk protein materials have various mechanical characteristics such as elastic modulus, ultimate strength and fracture toughness. While the considerable mechanical characteristics of the core crystalline regions of spider silk proteins at the atomistic scale have been investigated through several experimental techniques and computational studies, there is a lack of comparison between spider and silkworm fibroins in the atomistic scale. In this study, we investigated the differences between the mechanical characteristics of spider and silkworm fibroin structures by applying molecular dynamics and steered molecular dynamics. We found that serine amino acids in silkworm fibroins not only increased the number of hydrogen bonds, but also altered their structural characteristics and mechanical properties. PMID:26806791

  14. Identification and expression pattern of the chemosensory protein gene family in the silkworm, Bombyx mori.

    PubMed

    Gong, Da-Ping; Zhang, Hui-Jie; Zhao, Ping; Lin, Ying; Xia, Qing-You; Xiang, Zhong-Huai

    2007-03-01

    Insect chemosensory proteins (CSPs) as well as odorant-binding proteins (OBPs) have been supposed to transport hydrophobic chemicals to receptors on sensory neurons. Compared with OBPs, CSPs are expressed more broadly in various insect tissues. We performed a genome-wide analysis of the candidate CSP gene family in the silkworm. A total of 20 candidate CSPs, including 3 gene fragments and 2 pseudogenes, were characterized based on their conserved cysteine residues and their similarity to CSPs in other insects. Some of these genes were clustered in the silkworm genome. The gene expression pattern of these candidates was investigated using RT-PCR and microarray, and the results showed that these genes were expressed primarily in mature larvae and the adult moth, suggesting silkworm CSPs may be involved in development. The majority of silkworm CSP genes are expressed broadly in tissues including the antennae, head, thorax, legs, wings, epithelium, testes, ovaries, pheromone glands, wing disks, and compound eyes. PMID:17296501

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

    PubMed Central

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

    2011-01-01

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

  16. 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

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

    PubMed

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

    2016-04-01

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

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

    PubMed

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

    2015-01-01

    The vertebrate estrogens include 17-β-estradiol (E2), which has an analog in silkworm ovaries. In this study, the Bombyx mori vitellogenin gene (BmVg) was used as a biomarker to analyze the function of the E2 in silkworm. In most oviparous animals, Vg has female-specific expression. However, BmVg expression was also detected in B. mori males. Stage specific fluctuation of BmVg expression was similar in males and females, but expression levels in males were lower than in females. E2 treatment by injection or feeding of male larvae in the final instar stage induced and stimulated male BmVg transcription and protein synthesis. When silkworm ovary primordia were transplanted into males, BmVg was induced in male fat bodies. Transplanted ovaries primordia were also able to develop into ovaries and produce mature eggs. When females were treated with E2 promoted BmVg/BmVn protein accumulation in hemolymph, ovaries and eggs. However, BmVg transcription was decreased in female fat bodies. An E2 analog was identified in the hemolymph of day 3 wandering silkworms using high-performance liquid chromatography. Estradiol titers from fifth late-instar larvae to pupal stage were determined by enzyme-linked immunosorbent assay. The results suggested that silkworms synthesized a vertebrate E2 analog. This study found that E2 promoted the synthesis of BmVg, a female typical protein in silkworms. PMID:25285397

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

    PubMed

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

    2015-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2013-06-01

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

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed Central

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

    2014-01-01

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

  3. Genome-wide identification of abiotic stress-regulated and novel microRNAs in mulberry leaf.

    PubMed

    Wu, Ping; Han, Shaohua; Zhao, Weiguo; Chen, Tao; Zhou, Jiachun; Li, Long

    2015-10-01

    As the most important food plant for sericultural industry, mulberry trees have to suffer from a wide range of abiotic and biotic stresses, such as drought and high salinity. MicroRNAs (miRNAs) have been proved to play important roles in abiotic stresses regulation in many plants. However, there are seldom reports on the miRNAs expression profiles upon abiotic challenges in mulberry. In this study, three small RNA libraries from mulberry leaf tissue with or without drought or salt treatment were constructed and deep sequenced. Total of 48 conserved miRNAs (including miRNA*) and 162 novel miRNAs were identified (processing precision value>0.1). A total of 270 and 1963 target genes were predicted for conserved miRNAs and novel miRNAs, respectively. 13 differentially expressed miRNAs were detected under drought or salt stresses by deep sequencing and qRT-PCR. 5' RLM-RACE validated Morus 013341 to be the target gene of miR-395a. Our results provided initial clue to further study molecular mechanism on abiotic stresses regulation in mulberry. PMID:26188501

  4. Identification and Chacterization of new strains of Enterobacter spp. causing Mulberry (Morus alba) wilt disease in China

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new mulberry wilt disease (MWD) was recently identified in Hangzhou, Zhejiang province, China. Typical symptoms of the disease are dark brown discolorations in vascular tissues, leaf wilt, defoliation, and tree decline. Unlike the bacterial wilt disease caused by Ralstonia solanacearum, the leaf w...

  5. Proteome identification of the silkworm middle silk gland.

    PubMed

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

    2016-03-01

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

  6. 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.

  7. Antiviral substance from silkworm faeces: purification and its chemical characterization.

    PubMed

    Hiraki, A; Yukawa, M; Kim, J; Ueda, S

    1997-05-01

    In a previous paper, we reported that an extract of silkworm faeces has a marked antiviral activity on enveloped viruses, but not on a non-enveloped virus, and we showed that it inhibits the synthesis of a viral specific gene of HVJ (Sendai virus) without affecting the viral adsorption and entry into the host cell. In this paper, we accomplished the purification of an antiviral active substance by extraction of a hydrophobic substance and thin layer chromatography. The active substance was found to be a chlorophyll-like substance with a molecular mass of about 530. This substance shows clear antiviral activity against HVJ, HSV (herpes simplex virus type-1), and HIV (human immunodeficiency virus type-1). Its antiviral activity was dependent on light irradiation and temperature. Furthermore, it also possesses a strong hemolytic activity under light. PMID:9178937

  8. Proteome identification of the silkworm middle silk gland

    PubMed Central

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

    2016-01-01

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

  9. Precocious Metamorphosis in the Juvenile Hormone–Deficient Mutant of the Silkworm, Bombyx mori

    PubMed Central

    Daimon, Takaaki; Kozaki, Toshinori; Niwa, Ryusuke; Kobayashi, Isao; Furuta, Kenjiro; Namiki, Toshiki; Uchino, Keiro; Banno, Yutaka; Katsuma, Susumu; Tamura, Toshiki; Mita, Kazuei; Sezutsu, Hideki; Nakayama, Masayoshi; Itoyama, Kyo; Shimada, Toru; Shinoda, Tetsuro

    2012-01-01

    Insect molting and metamorphosis are intricately governed by two hormones, ecdysteroids and juvenile hormones (JHs). JHs prevent precocious metamorphosis and allow the larva to undergo multiple rounds of molting until it attains the proper size for metamorphosis. In the silkworm, Bombyx mori, several “moltinism” mutations have been identified that exhibit variations in the number of larval molts; however, none of them have been characterized molecularly. Here we report the identification and characterization of the gene responsible for the dimolting (mod) mutant that undergoes precocious metamorphosis with fewer larval–larval molts. We show that the mod mutation results in complete loss of JHs in the larval hemolymph and that the mutant phenotype can be rescued by topical application of a JH analog. We performed positional cloning of mod and found a null mutation in the cytochrome P450 gene CYP15C1 in the mod allele. We also demonstrated that CYP15C1 is specifically expressed in the corpus allatum, an endocrine organ that synthesizes and secretes JHs. Furthermore, a biochemical experiment showed that CYP15C1 epoxidizes farnesoic acid to JH acid in a highly stereospecific manner. Precocious metamorphosis of mod larvae was rescued when the wild-type allele of CYP15C1 was expressed in transgenic mod larvae using the GAL4/UAS system. Our data therefore reveal that CYP15C1 is the gene responsible for the mod mutation and is essential for JH biosynthesis. Remarkably, precocious larval–pupal transition in mod larvae does not occur in the first or second instar, suggesting that authentic epoxidized JHs are not essential in very young larvae of B. mori. Our identification of a JH–deficient mutant in this model insect will lead to a greater understanding of the molecular basis of the hormonal control of development and metamorphosis. PMID:22412378

  10. Ultrasound extraction of polysaccharides from mulberry leaves and their effect on enhancing antioxidant activity.

    PubMed

    Zhang, Dong-Yang; Wan, Yi; Xu, Jian-Yi; Wu, Guo-Hua; Li, Long; Yao, Xiao-Hui

    2016-02-10

    A Box-Behnken design (BBD) was applied to optimize the ultrasound-assisted extraction of polysaccharides from mulberry leaves. Under the optimum conditions of an extraction temperature of 57 °C, an extraction time of 80 min and a liquid/solid ratio of 53 mL/g, the mulberry leaf polysaccharide (MLP) yield was 6.92 ± 0.29%. Then, three fractions of MLPs were obtained by deproteinization, dialysis and decolorization. The carbohydrate content, FT-IR spectrum and monosaccharide composition of the MLPs were also investigated. The antioxidant activities of the three fractions were compared, and the results indicated that the antioxidant activities decreased with the increasing MLP purity. Therefore, highly concentrated MLPs were shown to have very little antioxidant activity. After quercetin (10 μg/mL) was added, the antioxidant activities were improved significantly. This result showed that MLPs and quercetin have a synergistic effect on the antioxidant activity. Although the MLPs have very little antioxidant activity alone, they greatly enhance the antioxidant activity of flavonoids. Thus, MLPs can be used as an antioxidant activity enhancer in the food industry. PMID:26686153

  11. Metabolic Effects of Mulberry Leaves: Exploring Potential Benefits in Type 2 Diabetes and Hyperuricemia

    PubMed Central

    Hunyadi, A.; Liktor-Busa, E.; Mrki, .; Martins, A.; Jedlinszki, N.; Hsieh, T. J.; Bthori, M.; Hohmann, J.; Zupk, I.

    2013-01-01

    The leaves of Morus alba L. have a long history in Traditional Chinese Medicine and also became valued by the ethnopharmacology of many other cultures. The worldwide known antidiabetic use of the drug has been suggested to arise from a complex combination effect of various constituents. Moreover, the drug is also a potential antihyperuricemic agent. Considering that type 2 diabetes and hyperuricemia are vice-versa in each other's important risk factors, the use of mulberry originated phytotherapeutics might provide an excellent option for the prevention and/or treatment of both conditions. Here we report a series of relevant in vitro and in vivo studies on the bioactivity of an extract of mulberry leaves and its fractions obtained by a stepwise gradient on silica gel. In vivo antihyperglycemic and antihyperuricemic activity, plasma antioxidant status, as well as in vitro glucose consumption by adipocytes in the presence or absence of insulin, xanthine oxidase inhibition, free radical scavenging activity, and inhibition of lipid peroxidation were tested. Known bioactive constituents of M. alba (chlorogenic acid, rutin, isoquercitrin, and loliolide) were identified and quantified from the HPLC-DAD fingerprint chromatograms. Iminosugar contents were investigated by MS/MS, 1-deoxynojirimycin was quantified, and amounts of 2-O-alpha-D-galactopyranosyl-1-deoxynojirimicin and fagomine were additionally estimated. PMID:24381639

  12. Adsorption properties of macroporous adsorbent resins for separation of anthocyanins from mulberry.

    PubMed

    Chen, Yao; Zhang, Weijie; Zhao, Ting; Li, Fang; Zhang, Min; Li, Jing; Zou, Ye; Wang, Wei; Cobbina, Samuel J; Wu, Xiangyang; Yang, Liuqing

    2016-03-01

    In this study, the adsorption/desorption characteristics of mulberry anthocyanins (MA) on five types of macroporous resins (XAD-7HP, AB-8, HP-20, D-101 and X-5) were evaluated, XAD-7HP and AB-8 showed higher adsorption/desorption capacities. On the basis of static adsorption test, XAD-7HP and AB-8 resins were selected for kinetics, isotherms and thermodynamics. The adsorption mechanism indicated that the process was better explained by pseudo-first-order kinetics and the Langmuir isotherm model, and the thermodynamics tests showed that the processes were exothermic, spontaneous and thermodynamically feasible. Dynamic tests were performed on a column packed with XAD-7HP and AB-8, and breakthrough volume was reached at 15 and 14 bed volumes of MA solution, respectively. The purity of the fraction by 40% ethanol elution on XAD-7HP reached 93.6%, from which cyanidin-3-glucoside and cyanidin-3-rutinoside were identified by HPLC-ESI-MS/MS. The method could be used to prepare high purity anthocyanins from mulberry fruits as well as other plants. PMID:26471611

  13. Antihemolytic Activities of Green Tea, Safflower, and Mulberry Extracts during Plasmodium berghei Infection in Mice

    PubMed Central

    Audomkasok, Suthin; Singpha, Waraporn; Chachiyo, Sukanya; Somsak, Voravuth

    2014-01-01

    Malaria-associated hemolysis is associated with mortality in adult patients. It has been speculated that oxidative stress and inflammation induced by malaria parasite are involved in its pathophysiology. Hence, we aimed to investigate the antihemolytic effect of green tea, safflower, and mulberry extracts against Plasmodium berghei infection. Aqueous crude extracts of these plants were prepared using hot water method and used for oral treatment in mice. Groups of ICR mice were infected with 6 × 106 infected red blood cells of P. berghei ANKA by intraperitoneal injection and given the extracts (500, 1500, and 3000 mg/kg) twice a day for 4 consecutive days. To assess hemolysis, hematocrit levels were then evaluated. Malaria infection resulted in hemolysis. However, antihemolytic effects were observed in infected mice treated with these extracts at dose-dependent manners. In conclusion, aqueous crude extracts of green tea, safflower, and mulberry exerted antihemolysis induced by malaria infection. These plants may work as potential source in the development of variety of herbal formulations for malarial treatment. PMID:25485155

  14. Characterization of bionanocomposite films prepared with agar and paper-mulberry pulp nanocellulose.

    PubMed

    Reddy, Jeevan Prasad; Rhim, Jong-Whan

    2014-09-22

    Crystallized nanocellulose (CNC) was separated from paper-mulberry (Broussonetia kazinoki Siebold) bast pulp by sulfuric acid hydrolysis method and they were blended with agar to prepare bionanocomposite films. The effect of CNC content (1, 3, 5 and 10 wt% based on agar) on the mechanical, water vapor permeability (WVP), and thermal properties of the nanocomposites were studied. Changes of the cellulose fibers in structure, morphology, crystallinity, and thermal properties of the films were evaluated using FT-IR, TEM, SEM, XRD, and TGA analysis methods. The CNC was composed of fibrous and spherical or elliptic granules of nano-cellulose with sizes of 50-60 nm. Properties of agar film such as mechanical and water vapor barrier properties were improved significantly (p<0.05) by blending with the CNC. The tensile modulus and tensile strength of agar film increased by 40% and 25%, respectively, in the composite film with 5 wt% of CNC, and the WVP of agar film decreased by 25% after formation of nanocomposite with 3 wt% of CNC. The CNC obtained from the paper-mulberry bast pulp can be used as a reinforcing agent for the preparation of bio-nanocomposites, and they have a high potential for the development of completely biodegradable food packaging materials. PMID:24906782

  15. Definition of Eight Mulberry Species in the Genus Morus by Internal Transcribed Spacer-Based Phylogeny

    PubMed Central

    Zeng, Qiwei; Chen, Hongyu; Zhang, Chao; Han, Minjing; Li, Tian; Qi, Xiwu; Xiang, Zhonghuai; He, Ningjia

    2015-01-01

    Mulberry, belonging to the order Rosales, family Moraceae, and genus Morus, has received attention because of both its economic and medicinal value, as well as for its important ecological function. The genus Morus has a worldwide distribution, however, its taxonomy remains complex and disputed. Many studies have attempted to classify Morus species, resulting in varied numbers of designated Morus spp. To address this issue, we used information from internal transcribed spacer (ITS) genetic sequences to study the taxonomy of all the members of generally accepted genus Morus. We found that intraspecific 5.8S rRNA sequences were identical but that interspecific 5.8S sequences were diverse. M. alba and M. notabilis showed the shortest (215 bp) and the longest (233 bp) ITS1 sequence length, respectively. With the completion of the mulberry genome, we could identify single nucleotide polymorphisms within the ITS locus in the M. notabilis genome. From reconstruction of a phylogenetic tree based on the complete ITS data, we propose that the Morus genus should be classified into eight species, including M. alba, M. nigra, M. notabilis, M. serrata, M. celtidifolia, M. insignis, M. rubra, and M. mesozygia. Furthermore, the classification of the ITS sequences of known interspecific hybrid clones into both paternal and maternal clades indicated that ITS variation was sufficient to distinguish interspecific hybrids in the genus Morus. PMID:26266951

  16. Definition of Eight Mulberry Species in the Genus Morus by Internal Transcribed Spacer-Based Phylogeny.

    PubMed

    Zeng, Qiwei; Chen, Hongyu; Zhang, Chao; Han, Minjing; Li, Tian; Qi, Xiwu; Xiang, Zhonghuai; He, Ningjia

    2015-01-01

    Mulberry, belonging to the order Rosales, family Moraceae, and genus Morus, has received attention because of both its economic and medicinal value, as well as for its important ecological function. The genus Morus has a worldwide distribution, however, its taxonomy remains complex and disputed. Many studies have attempted to classify Morus species, resulting in varied numbers of designated Morus spp. To address this issue, we used information from internal transcribed spacer (ITS) genetic sequences to study the taxonomy of all the members of generally accepted genus Morus. We found that intraspecific 5.8S rRNA sequences were identical but that interspecific 5.8S sequences were diverse. M. alba and M. notabilis showed the shortest (215 bp) and the longest (233 bp) ITS1 sequence length, respectively. With the completion of the mulberry genome, we could identify single nucleotide polymorphisms within the ITS locus in the M. notabilis genome. From reconstruction of a phylogenetic tree based on the complete ITS data, we propose that the Morus genus should be classified into eight species, including M. alba, M. nigra, M. notabilis, M. serrata, M. celtidifolia, M. insignis, M. rubra, and M. mesozygia. Furthermore, the classification of the ITS sequences of known interspecific hybrid clones into both paternal and maternal clades indicated that ITS variation was sufficient to distinguish interspecific hybrids in the genus Morus. PMID:26266951

  17. Specificity determinants of the silkworm moth sex pheromone.

    PubMed

    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

  18. Improvement of glycosylation structure by suppression of β-N-acetylglucosaminidases in silkworm.

    PubMed

    Nomura, Tsuyoshi; Suganuma, Masatoshi; Higa, Yukiko; Kataoka, Yukiko; Funaguma, Shunsuke; Okazaki, Hironobu; Suzuki, Takeo; Kobayashi, Isao; Sezutsu, Hideki; Fujiyama, Kazuhito

    2015-02-01

    The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. However, the presence of paucimannosidic-type N-glycan in glycoproteins restricts their clinical use. Paucimannosidic-type N-glycan is produced by insect-specific membrane-binding-type β-N-acetylglucosaminidase (GlcNAcase). In the silkworm, BmGlcNAcase1, BmGlcNAcase2, and BmFDL are membrane-binding-type GlcNAcases. We investigated the localization of these GlcNAcases and found that BmFDL and BmGlcNAcase2 were mainly located in the fat body and hemolymph, respectively. The fat body is the main tissue of recombinant protein expression by baculovirus, and many glycoproteins are secreted into the hemolymph. These results suggest that inhibition of BmFDL and BmGlcNAcase2 could increase GlcNAc-type N-glycan levels. We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). Next, we generated a transgenic silkworm in which RNA interference (RNAi) reduced the BmFDL transcript level and enzyme activity to 25% and 50%, respectively, of that of the control silkworm. The proportion of GlcNAc-type N-glycan increased to 4.3% in the RNAi-transgenic silkworm. We conclude that the structure of N-glycan can be changed by inhibiting the GlcNAcases in silkworm. PMID:25193875

  19. Large-scale intersubspecific recombination in the plant-pathogenic bacterium Xylella fastidiosa is associated with the host shift to mulberry.

    PubMed

    Nunney, Leonard; Schuenzel, Erin L; Scally, Mark; Bromley, Robin E; Stouthamer, Richard

    2014-05-01

    Homologous recombination plays an important role in the structuring of genetic variation of many bacteria; however, its importance in adaptive evolution is not well established. We investigated the association of intersubspecific homologous recombination (IHR) with the shift to a novel host (mulberry) by the plant-pathogenic bacterium Xylella fastidiosa. Mulberry leaf scorch was identified about 25 years ago in native red mulberry in the eastern United States and has spread to introduced white mulberry in California. Comparing a sequence of 8 genes (4,706 bp) from 21 mulberry-type isolates to published data (352 isolates representing all subspecies), we confirmed previous indications that the mulberry isolates define a group distinct from the 4 subspecies, and we propose naming the taxon X. fastidiosa subsp. morus. The ancestry of its gene sequences was mixed, with 4 derived from X. fastidiosa subsp. fastidiosa (introduced from Central America), 3 from X. fastidiosa subsp. multiplex (considered native to the United States), and 1 chimeric, demonstrating that this group originated by large-scale IHR. The very low within-type genetic variation (0.08% site polymorphism), plus the apparent inability of native X. fastidiosa subsp. multiplex to infect mulberry, suggests that this host shift was achieved after strong selection acted on genetic variants created by IHR. Sequence data indicate that a single ancestral IHR event gave rise not only to X. fastidiosa subsp. morus but also to the X. fastidiosa subsp. multiplex recombinant group which infects several hosts but is the only type naturally infecting blueberry, thus implicating this IHR in the invasion of at least two novel native hosts, mulberry and blueberry. PMID:24610840

  20. Specific Detection and Identification of American Mulberry-Infecting and Italian Olive-Associated Strains of Xylella fastidiosa by Polymerase Chain Reaction

    PubMed Central

    Guan, Wei; Shao, Jonathan; Elbeaino, Toufic; Davis, Robert E.; Zhao, Tingchang; Huang, Qi

    2015-01-01

    Xylella fastidiosa causes bacterial leaf scorch in many landscape trees including elm, oak, sycamore and mulberry, but methods for specific identification of a particular tree host species-limited strain or differentiation of tree-specific strains are lacking. It is also unknown whether a particular landscape tree-infecting X. fastidiosa strain is capable of infecting multiple landscape tree species in an urban environment. We developed two PCR primers specific for mulberry-infecting strains of X. fastidiosa based on the nucleotide sequence of a unique open reading frame identified only in mulberry-infecting strains among all the North and South American strains of X. fastidiosa sequenced to date. PCR using the primers allowed for detection and identification of mulberry-infecting X. fastidiosa strains in cultures and in samples collected from naturally infected mulberry trees. In addition, no mixed infections with or non-specific detections of the mulberry-infecting strains of X. fastidiosa were found in naturally X. fastidiosa-infected oak, elm and sycamore trees growing in the same region where naturally infected mulberry trees were grown. This genotype-specific PCR assay will be valuable for disease diagnosis, studies of strain-specific infections in insects and plant hosts, and management of diseases caused by X. fastidiosa. Unexpectedly but interestingly, the unique open reading frame conserved in the mulberry-infecting strains in the U. S. was also identified in the recently sequenced olive-associated strain CoDiRO isolated in Italy. When the primer set was tested against naturally infected olive plant samples collected in Italy, it allowed for detection of olive-associated strains of X. fastidiosa in Italy. This PCR assay, therefore, will also be useful for detection and identification of the Italian group of X. fastidiosa strains to aid understanding of the occurrence, evolution and biology of this new group of X. fastidiosa strains. PMID:26061051

  1. Specific Detection and Identification of American Mulberry-Infecting and Italian Olive-Associated Strains of Xylella fastidiosa by Polymerase Chain Reaction.

    PubMed

    Guan, Wei; Shao, Jonathan; Elbeaino, Toufic; Davis, Robert E; Zhao, Tingchang; Huang, Qi

    2015-01-01

    Xylella fastidiosa causes bacterial leaf scorch in many landscape trees including elm, oak, sycamore and mulberry, but methods for specific identification of a particular tree host species-limited strain or differentiation of tree-specific strains are lacking. It is also unknown whether a particular landscape tree-infecting X. fastidiosa strain is capable of infecting multiple landscape tree species in an urban environment. We developed two PCR primers specific for mulberry-infecting strains of X. fastidiosa based on the nucleotide sequence of a unique open reading frame identified only in mulberry-infecting strains among all the North and South American strains of X. fastidiosa sequenced to date. PCR using the primers allowed for detection and identification of mulberry-infecting X. fastidiosa strains in cultures and in samples collected from naturally infected mulberry trees. In addition, no mixed infections with or non-specific detections of the mulberry-infecting strains of X. fastidiosa were found in naturally X. fastidiosa-infected oak, elm and sycamore trees growing in the same region where naturally infected mulberry trees were grown. This genotype-specific PCR assay will be valuable for disease diagnosis, studies of strain-specific infections in insects and plant hosts, and management of diseases caused by X. fastidiosa. Unexpectedly but interestingly, the unique open reading frame conserved in the mulberry-infecting strains in the U. S. was also identified in the recently sequenced olive-associated strain CoDiRO isolated in Italy. When the primer set was tested against naturally infected olive plant samples collected in Italy, it allowed for detection of olive-associated strains of X. fastidiosa in Italy. This PCR assay, therefore, will also be useful for detection and identification of the Italian group of X. fastidiosa strains to aid understanding of the occurrence, evolution and biology of this new group of X. fastidiosa strains. PMID:26061051

  2. Repression of tyrosine hydroxylase is responsible for the sex-linked chocolate mutation of the silkworm, Bombyx mori.

    PubMed

    Liu, Chun; Yamamoto, Kimiko; Cheng, Ting-Cai; Kadono-Okuda, Keiko; Narukawa, Junko; Liu, Shi-Ping; Han, Yu; Futahashi, Ryo; Kidokoro, Kurako; Noda, Hiroaki; Kobayashi, Isao; Tamura, Toshiki; Ohnuma, Akio; Banno, Yutaka; Dai, Fang-Ying; Xiang, Zhong-Huai; Goldsmith, Marian R; Mita, Kazuei; Xia, Qing-You

    2010-07-20

    Pigmentation patterning has long interested biologists, integrating topics in ecology, development, genetics, and physiology. Wild-type neonatal larvae of the silkworm, Bombyx mori, are completely black. By contrast, the epidermis and head of larvae of the homozygous recessive sex-linked chocolate (sch) mutant are reddish brown. When incubated at 30 degrees C, mutants with the sch allele fail to hatch; moreover, homozygous mutants carrying the allele sch lethal (sch(l)) do not hatch even at room temperature (25 degrees C). By positional cloning, we narrowed a region containing sch to 239,622 bp on chromosome 1 using 4,501 backcross (BC1) individuals. Based on expression analyses, the best sch candidate gene was shown to be tyrosine hydroxylase (BmTh). BmTh coding sequences were identical among sch, sch(l), and wild-type. However, in sch the approximately 70-kb sequence was replaced with approximately 4.6 kb of a Tc1-mariner type transposon located approximately 6 kb upstream of BmTh, and in sch(l), a large fragment of an L1Bm retrotransposon was inserted just in front of the transcription start site of BmTh. In both cases, we observed a drastic reduction of BmTh expression. Use of RNAi with BmTh prevented pigmentation and hatching, and feeding of a tyrosine hydroxylase inhibitor also suppressed larval pigmentation in the wild-type strain, pnd(+) and in a pS (black-striped) heterozygote. Feeding L-dopa to sch neonate larvae rescued the mutant phenotype from chocolate to black. Our results indicate the BmTh gene is responsible for the sch mutation, which plays an important role in melanin synthesis producing neonatal larval color. PMID:20615980

  3. Power ultrasound as a pretreatment to convective drying of mulberry (Morus alba L.) leaves: Impact on drying kinetics and selected quality properties.

    PubMed

    Tao, Yang; Wang, Ping; Wang, Yilin; Kadam, Shekhar U; Han, Yongbin; Wang, Jiandong; Zhou, Jianzhong

    2016-07-01

    The effect of ultrasound pretreatment prior to convective drying on drying kinetics and selected quality properties of mulberry leaves was investigated in this study. Ultrasound pretreatment was carried out at 25.2-117.6W/L for 5-15min in a continuous mode. After sonication, mulberry leaves were dried in a hot-air convective dryer at 60°C. The results revealed that ultrasound pretreatment not only affected the weight of mulberry leaves, it also enhanced the convective drying kinetics and reduced total energy consumption. The drying kinetics was modeled using a diffusion model considering external resistance and effective diffusion coefficient De and mass transfer coefficient hm were identified. Both De and hm during convective drying increased with the increase of acoustic energy density (AED) and ultrasound duration. However, De and hm increased slowly at high AED levels. Furthermore, ultrasound pretreatment had a more profound influence on internal mass transfer resistance than on external mass transfer resistance during drying according to Sherwood numbers. Regarding the quality properties, the color, antioxidant activity and contents of several bioactive compounds of dried mulberry leaves pretreated by ultrasound at 63.0W/L for 10min were similar to that of mulberry leaves without any pretreatments. Overall, ultrasound pretreatment is effective to shorten the subsequent drying time of mulberry leaves without damaging the quality of final product. PMID:26964954

  4. In Vivo Hypoglycaemic Effect and Inhibitory Mechanism of the Branch Bark Extract of the Mulberry on STZ-Induced Diabetic Mice

    PubMed Central

    Liu, Hua-Yu; Fang, Meng; Zhang, Yu-Qing

    2014-01-01

    Branch bark extract (BBE) derived from the mulberry cultivar Husang 32 (Morus multicaulis L.) with aqueous alcohol solution has been investigated as an inhibitor of α-glycosidase in vitro. Mulberry BBE was orally administered to STZ-induced diabetic mice for three weeks, and it improved the weight gain and ameliorated the swelling of liver and kidney in diabetic mice. Obviously, mulberry BBE not only can reduce the abnormally elevated levels of serum insulin and ameliorate insulin resistance induced by STZ, but also it regulates dyslipidemia in diabetic mice. To understand this therapeutic effect and the regulatory mechanisms of BBE in diabetic mice, a qRT-PCR experiment was performed, indicating that the mulberry BBE can regulate the mRNA expression of glycometabolism genes in diabetic mice, including glucose-6-phosphatase (G6Pase), glucokinase (GCK), and phosphoenolpyruvate carboxykinase (PEPCK), thereby regulating sugar metabolism and reducing the blood glucose level in diabetic mice. The mulberry BBE can increase the mRNA expression of the genes Ins1, Ins2 and pancreatic duodenal homeobox-1 (PDX-1) and may decrease the insulin resistance in diabetic mice. Those results provide an important basis for making the best use of mulberry branch resources and producing biomedical drugs with added value. PMID:25177729

  5. Simple, selective, and rapid quantification of 1-deoxynojirimycin in mulberry leaf products by high-performance anion-exchange chromatography with pulsed amperometric detection.

    PubMed

    Yoshihashi, Tadashi; Do, Huong Thi Thu; Tungtrakul, Patcharee; Boonbumrung, Sumitra; Yamaki, Kohji

    2010-04-01

    1-Deoxynojirimycin (DNJ) occurs in mulberry and other plants and is a highly potent glycosidase inhibitor reported to suppress blood glucose levels, thus preventing diabetes. Derivatization is required for quantification of DNJ upon use of spectral detection methods. Because of this difficulty, the DNJ contents of mulberry-based food products are rarely stated, even if DNJ is their active component. A simple, selective, and rapid method of high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) to quantify DNJ in mulberry-based food products was developed. Stability testing of DNJ under heat treatment was also performed. A water extract of mulberry tea sample was subjected to HPAEC-PAD in a CarboPac MA1 column with a sodium hydroxide gradient. DNJ was clearly separated at a retention time of 7.26 min without interference and was selectively detected in the water extract. The detection limit was 5 ng. Heat stability studies suggested that DNJ was heat stable. HPAEC-PAD was not subject to interference, was highly selective for DNJ, and was superior to other high-performance liquid chromatography (HPLC) techniques in terms of sample preparation, resolution, and sensitivity. The method allowed simple, selective, and rapid analysis of DNJ in food matrices and might be useful for development of mulberry-based food products. Heat treatment could be an option for sterilizing mulberry-based products. PMID:20492274

  6. Dosage analysis of Z chromosome genes using microarray in silkworm, Bombyx mori.

    PubMed

    Zha, Xingfu; Xia, Qingyou; Duan, Jun; Wang, Chunyun; He, Ningjia; Xiang, Zhonghuai

    2009-01-01

    In many organisms, dosage compensation is needed to equalize sex-chromosome gene expression in males and females. Several genes on silkworm Z chromosome were previously detected to show a higher expression level in males and lacked dosage compensation. Whether silkworm lacks global dosage compensation still remains poorly known. Here, we analyzed male:female (M:F) ratios of expression of chromosome-wide Z-linked genes in the silkworm using microarray data. The expression levels of genes on Z chromosome in each tissue were significantly higher in males compared to females, which indicates no global dosage compensation in silkworm. Interestingly, we also found some genes with no bias (M:F ratio: 0.8-1.2) on the Z chromosome. Comparison of male-biased (M:F ratio more than 1.5) and unbiased genes indicated that the two sets of the genes have functional differences. Analysis of gene expression by sex showed that M:F ratios were, to some extent, associated with their expression levels. These results provide useful clues to further understanding roles of dosage of Z chromosome and some Z-linked sexual differences in silkworms. PMID:19150406

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

    PubMed

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

    2014-10-10

    Female qualities in males are common in vertebrates but have not been extensively reported in insects. Vitellogenin (Vg) is highly expressed in the female fat body and is generally required for the formation of yolk proteins in the insect egg. Vg upregulation is generally regarded as a female quality in female oviparous animals. In this study, we found that Bombyx mori Vg (BmVg) is especially highly expressed in the female pupa. Downregulation of the BmVg gene in the female pupa by RNA interference (RNAi) interfered with egg formation and embryonic development, showing the importance of BmVg in these processes. So, we used BmVg as a biomarker for female qualities in the silkworm. Hematoxylin-eosin staining and immunofluorescence histochemistry showed that ovary transplants induced BmVg synthesis in the male pupa fat body. Ovaries transplanted into male silkworms produced only a few eggs with deformed yolk granules. These results suggested that the amount of BmVg in the male silkworm was insufficient for eggs to undergo complete embryonic development. After 17-beta-estradiol was used to treat male pupae and male pupal fat bodies, BmVg was upregulated in vivo and in vitro. These findings indicated that the male silkworm has innate female qualities that were induced by a transplanted ovary and 17β-estradiol. However, in silkworms, female qualities in males are not as complete as in females. PMID:25242521

  8. Silkworm pupae powder ingestion increases fat metabolism in swim-trained rats

    PubMed Central

    Ryu, Sung Pil

    2014-01-01

    [Purpose] Many researchers are trying to solve the metabolic syndrome by utilizing a variety of nutritional control and exercise. Of those, silkworm pupae peptides are known to inhibit the synthesis of fat. Therefore, we examine the effect of fat metabolism by supplying silkworm pupae (SP) for 5-week in swim-trained rats. [Methods] Animals were divided into four groups as a group (n = 32) fed a normal diet (CO) with exercise training (CE); a group fed a silkworm pupa diet (SPC) with an exercise training (SPE), respectively. [Results] Abdominal fat pads (abdominal and epididymal) weight were lowest in SPE. The serum triglyceride, total cholesterol concentrations were lower in the SP and the SPE. HDL-cholesterol, however, was not different between groups. Liver AMPK (AMP-activated protein kinase) was increased in the CE and the SPE. Liver PPAR-α (Peroxisome proliferator-activated receptor alpha) was increased in the SPC and SPE. L-FABP (liver fatty acids binding protein) was increased by SP ingestion. Liver CPT-1 (carnitine palmitoyltransferase-1) protein expression was increased by exercise training only. [Conclusion] In the present study showed that the silkworm pupae intake and/or swimming exercise training activates fat metabolism to reduce the concentration of serum lipids. Thus, the silkworm pupae intake leads to a reduction in fat storage, this is considered to be effective in the inhibition of the metabolic syndrome. PMID:25566449

  9. Spatiotemporal expression profile of the Pumilio gene in the embryonic development of silkworm.

    PubMed

    Chen, Liang; You, Zaizhi; Xia, Hengchuan; Tang, Qi; Zhou, Yang; Yao, Qin; Chen, Keping

    2014-01-01

    We previously identified a pumilio gene in silkworm (Bombyx mori L.), designated BmPUM, which was specifically expressed in the ovary and testis. To further characterize this gene's involvement in silkworm development, we have determined the spatiotemporal expression pattern of BmPUM during all embryonic stages. Real-time polymerase chain reaction (RT-PCR) analysis revealed that BmPUM was expressed in all stages of silkworm embryos and that its transcript levels displayed two distinct peaks. The first was observed at the germ-band formation stage (1 d after oviposition) and dropped to a low level at the gonad formation stage (5 d after oviposition). The second was detected at the stage of bristle follicle occurrence (6 d after oviposition), which was confirmed by Western blot analysis and immunohistochemistry. Nanos (Nos), functioning together with Pum in abdomen formation of Drosophila embryos, was also highly expressed at the beginning (0 h to 1 d after oviposition) of embryogenesis, but its transcript levels were very low after the stage of germ-band formation. These results suggest that BmPUM functions with Bombyx mori nanos (Bm-nanos) at the early stages of silkworm embryonic development, and may then play a role in gonad formation and the occurrence of bristle follicles. Our data thus provide a foundation to uncover the role of BmPUM during silkworm development. PMID:25265852

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

    PubMed

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

    2014-12-01

    The nuclear receptor, ultraspiracle protein (USP), is a transcription factor and an essential component of a heterodimeric receptor complex with ecdysone receptor. However, the mechanisms underlying the transcriptional regulation of USP in silkworm are unknown. In this study, using dual-spike-in qPCR method, we examined the expression of Bombyx ultraspiracle gene (BmUSP) in various tissues of silkworm as well as expression changes after stimulation with ecdysone. The results showed that the expression levels of BmUSP gene varied in different tissues and were increased 2 h after exposure to ecdysone. To identify the molecular mechanism underlying the regulation of USP gene expression in silkworm Bombyx mori, promoter truncation analyses were performed using the luciferase reporter assay and Bac-to-Bac expression system in several tissues of B. mori. BmUSP gene promoter with 5' end serial deletions showed different levels of activity in various tissues, higher in fat body and Malpighian tubule. Deletion of the region from -485 to -445 and -307 to -281 upstream of BmUSP gene abolished and increased its promoter activity, respectively. This region contains AP-1, Dfd transcription factor binding sites. These results indicate that BmUSP are expressed at different levels in different tissues of the silkworm, but all are subjected to the regulation by ecdysone. This study would provide an important foundation for investigating the mechanism underlying the transcriptional regulation of BmUSP in the silkworm. PMID:25175165

  11. 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 μg 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

  12. In vitro and In vivo Antioxidant Activity of Flavonoid Extracted from Mulberry Fruit (Morus alba L.)

    PubMed Central

    Raman, Sivakumar Thasma; Ganeshan, Ajay Krishna Palani Gounder; Chen, Cheng; Jin, Chao; Li, Shao-Hui; Chen, Hui-Juan; Gui, Zhongzheng

    2016-01-01

    Background: Many plants possess antioxidants that exhibit additive or synergistic activities. Objective: In this study, an ethanol-extracted flavonoid extracted from mulberry fruit (FEM) was evaluated for the antioxidant activity in vitro and the hemolysis in red blood cell (RBC) and lipid peroxidation in liver in vivo. Materials and Methods: Antioxidant activities in vitro were measured by quantifying its 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity, reducing power, and Fe2+-chelating ability. FEM inhibits hemolysis in RBCs and effects of lipid peroxidation in the liver were estimated. Results: The total content of flavonoid compounds was 187.23 mg of quercetin equivalents per grams dried material. In the in vitro assays, FEM demonstrated a strong antioxidant effect, especially in DPPH scavenging activity and reducing power. Mouse RBC hemolysis induced by H2O2 was significantly inhibited by FEM in a dose- and time-dependent manner. The effects of FEM on lipid peroxidation in liver, mitochondria, and microsome were investigated. The percentage of inhibition at high concentration (100 μg/mL) of FEM was 45.51%, 39.36%, and 42.78% for liver, mitochondria, and microsomes, respectively. These results suggest that the FEM possesses a strong antioxidant activity both in vivo and in vitro. SUMMARY The total content of flavonoid compounds in mulberry fruit was 187.23 mg/g dried materialFEM showed a strong antioxidant effect, especially in 2,2-diphenyl-1-picrylhydrazyl scavenging activity and reducing powerMouse red blood cell hemolysis induced by H2O2 was significantly inhibited by FEM in a dose- and time-dependent mannerThe inhibition percentage at high concentration of FEM was 45.51%, 39.36%, and 42.78% for mouse’s liver, mitochondrial, and microsomes, respectively. Abbreviations used: FEM: Flavonoid Extracted from Mulberry fruit, H2O2: Hydrogen peroxide, DPPH: 2,2-diphenyl-1-picrylhydrazyl, EDTA: Ethylene diamine tetraacetic acid, MDA: malondialdehyde, TBA: 2-thiobarbituric acid, RBC: Red blood cells, DNJ: 1-deoxynojirimycin, LDL: low density lipoprotein, ROS: reactive oxygen species, EDTA2Na: Ethylenediaminetetraacetic acid disodium salt. PMID:27076749

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

    PubMed

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

    2005-01-21

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

  14. Radon flux measurements on Gardinier and Royster phosphogypsum piles near Tampa and Mulberry, Florida

    SciTech Connect

    Hartley, J.N.; Freeman, H.D.

    1986-01-01

    As part of the planned Environmental Protection Agency (EPA) radon flux monitoring program for the Florida phosphogypsum piles, Pacific Northwest Laboratory (PNL), under contract to the EPA, constructed 50 large-area passive radon collection devices and demonstrated their use at two phosphogypsum piles near Tampa and Mulberry, Florida. The passive devices were also compared to the PNL large-area flow-through system. The main objectives of the field tests were to demonstrate the use of the large-area passive radon collection devices to EPA and PEI personnel and to determine the number of radon flux measurement locations needed to estimate the average radon flux from a phosphogypsum pile. This report presents the results of the field test, provides recommendations for long-term monitoring, and includes a procedure for making the radon flux measurements.

  15. Value added of mulberry paper waste by carboxymethylation for preparation a packaging film

    NASA Astrophysics Data System (ADS)

    Rachtanapun, P.; Kumthai, S.; Mulkarat, N.; Pintajam, N.; Suriyatem, R.

    2015-07-01

    Cellulose from mulberry paper waste was converted to carboxymethyl cellulose (CMCm) by etherification using chloroacetic acid and various sodium hydroxide (NaOH) concentrations (30-60 g/100 mL). The degree of substitution of the various CMCm materials produced ranged between 0.33-0.45. The chemical structure of cellulose and the synthesized CMCm materials was characterized using Fourier transform infrared spectroscopy. CMCm films were cast and tested. The tensile strength (TS) and percent elongation at break (EB) of the films were investigated. The highest TS (32.58 MPa) and EB (2.39%) were found using the 50g /100 mL NaOH-synthesized CMCm film. The effect of glycerol as a plasticizer on TS and EB of this CMCm film was also evaluated. Increasing the glycerol content decreased TS and increased EB of the CMCm films.

  16. The complete chloroplast genome sequence of the mulberry Morus notabilis (Moreae).

    PubMed

    Chen, Chen; Zhou, Wen; Huang, Ying; Wang, Zhe-Zhi

    2016-07-01

    The complete chloroplast genome of the mulberry Morus notabilis (Moreae) has been reconstructed from the whole-genome Illumina sequencing data. The circular genome is 158,680 bp in size, and comprises a pair of inverted repeat (IR) regions of 25,717 bp each, a large single-copy (LSC) region of 87,470 bp and a small single-copy (SSC) region of 19,776 bp. The total A+T content is 63.6%, while the corresponding values of the LSC, SSC and IR region are 65.9%, 70.7% and 57.1%, respectively. The chloroplast genome contains 129 genes, including 84 protein-coding genes (PCGs), eight ribosomal RNA (rRNA) genes and 37 transfer RNA (tRNA) genes. The maximum likelihood (ML) phylogenetic analysis revealed that M. notabilis was more related to its congeners than to the others. PMID:26119116

  17. Complete mitochondrial genome of a hybrid strain of the domesticated silkworm (Qiufeng × Baiyu).

    PubMed

    Li, Fengbo; Zhang, Huixian; Liu, Peigang; Wang, Yongqiang; Meng, Zhiqi

    2016-05-01

    The hybrid strain of the domesticated silkworm (Qiufeng × Baiyu) is one of the most popular commercial silkworm varieties in China. In this study, we reported its complete mitochondrial genome sequence for the first time. The 15,680 bp long genome contains 37 genes (13 protein-coding genes [PCGs], 2 rRNA genes, and 22 tRNA genes) and 1 major non-coding A + T-rich region, with the typical arrangement found in Lepidoptera. All PCGs started with typical ATN codons except for COI, which began with CGA. Eleven PCGs have complete stop codons, whereas COI and COII end with a single T. The 495 bp long A + T-rich region harbors the conserved sequence features typically found in lepidopteran insects. The complete mitochondrial genome sequence of Qiufeng × Baiyu provides an important data source for further study on the mechanism of silkworm domestication. PMID:25319289

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

    PubMed

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

    2015-10-01

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

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

    PubMed Central

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

    2015-01-01

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

  1. 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

  2. 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.

  3. 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.

  4. [Mapping of the lethal genes in the sex-linkaged balanced lethal silkworm Bombyx mori using SSR markers].

    PubMed

    Xuan, Nan; Niu, Bao-Long; Wang, Hai-Long; Zhuang, Li; Meng, Zhi-Qi

    2010-12-01

    The males of sex-linkaged balanced lethal silkworm strain (S-14) has two non-allelic recessive genes (lethal gene 1, l1 and lethal gene 2, l2). The two genes are located on two different Z chromosomes and cause death of embryos at body pigmentation stage and end reversal embryo stage, respectively. We firstly hybridized the males of S-14 strain with the females having wild-type genes of P50 strain and then backcrossed the males of F1 with females of P50 strain. A total of 1660 female moths of BC1 generation were divided into two groups, 1100 in BC1-l1 and 560 in BC1-l2 according to the lethal gene carried by these female moths' fathers-fame moths of F1, respectively. Based on the nucleotide sequence information from the published physical map of Bombyx mori, we developed 16 polymorphic SSR markers in l1 gene region and 18 polymorphic SSR makers in l2 gene region compared to the allelic region of P50 strain and used these SSR markers and groups of BC1-l1 and BC1-12 to map the two lethal genes, respectively. Gene l1 was mapped on the region of Z chromosome, covering a physical distance of 2.60 Mb. Gene l2 was fine mapped on the region of Z chromosome, covering a physical distance of 0.69 Mb. PMID:21513153

  5. Transcription activator-like effector nuclease (TALEN)-mediated female-specific sterility in the silkworm, Bombyx mori.

    PubMed

    Xu, J; Wang, Y; Li, Z; Ling, L; Zeng, B; James, A A; Tan, A; Huang, Y

    2014-12-01

    Engineering sex-specific sterility is critical for developing transgene-based sterile insect technology. Targeted genome engineering achieved by customized zinc-finger nuclease, transcription activator-like effector nuclease (TALEN) or clustered, regularly interspaced, short palindromic repeats/Cas9 systems has been exploited extensively in a variety of model organisms; however, screening mutated individuals without a detectable phenotype is still challenging. In addition, genetically recessive mutations only detectable in homozygotes make the experiments time-consuming. In the present study, we model a novel genetic system in the silkworm, Bombyx mori, that results in female-specific sterility by combining transgenesis with TALEN technologies. This system induces sex-specific sterility at a high efficiency by targeting the female-specific exon of the B. mori doublesex (Bmdsx) gene, which has sex-specific splicing isoforms regulating somatic sexual development. Transgenic animals co-expressing TALEN left and right arms targeting the female-specific Bmdsx exon resulted in somatic mutations and female mutants lost fecundity because of lack of egg storage and abnormal external genitalia. The wild-type sexual dimorphism of abdominal segment was not evident in mutant females. In contrast, there were no deleterious effects in mutant male moths. The current somatic TALEN technologies provide a promising approach for future insect functional genetics, thus providing the basis for the development of attractive genetic alternatives for insect population management. PMID:25125145

  6. Analysis of Transcripts Expressed in One-Day-Old Larvae and Fifth Instar Silk Glands of Tasar Silkworm, Antheraea mylitta

    PubMed Central

    Maity, Samita; Goel, Sagar I.; Roy, Sobhan; Ghorai, Suvankar; Bhattacharyya, Swati; Venugopalan, Aravind; Ghosh, Ananta K.

    2010-01-01

    Antheraea mylitta is one of the wild nonmulberry silkworms, which produces tasar silk. An EST project has been undertaken to understand the gene expression profile of A. mylitta silk gland. Two cDNA libraries, one from the whole bodies of one-day-old larvae and the other from the silkglands of fifth instar larvae, were constructed and sequenced. A total of 2476 good-quality ESTs (1239 clones) were obtained and grouped into 648 clusters containing 390 contigs and 258 singletons to represent 467 potential unigenes. Forty-five sequences contained putative coding region, and represented potentially novel genes. Among the 648 clusters, 241 were categorized according to Gene Ontology hierarchy and showed presence of several silk and immune-related genes. The A. mylitta ESTs have been organized into a freely available online database “AmyBASE”. These data provide an initial insight into the A. mylitta transcriptome and help to understand the molecular mechanism of silk protein production in a Lepidopteran species. PMID:20454581

  7. Yellow-e Determines the Color Pattern of Larval Head and Tail Spots of the Silkworm Bombyx mori*

    PubMed Central

    Ito, Katsuhiko; Katsuma, Susumu; Yamamoto, Kimiko; Kadono-Okuda, Keiko; Mita, Kazuei; Shimada, Toru

    2010-01-01

    Yellow proteins form a large family in insects. In Drosophila melanogaster, there are 14 yellow genes in the genome. Previous studies have shown that the yellow gene is necessary for normal pigmentation; however, the roles of other yellow genes in body coloration are not known. Here, we provide the first evidence that yellow-e is required for normal body color pattern in insect larvae. In two mutant strains, bts and its allele bts2, of the silkworm Bombyx mori, the larval head cuticle and anal plates are reddish brown instead of the white color found in the wild type. Positional cloning revealed that deletions in the Bombyx homolog of the Drosophila yellow-e gene (Bmyellow-e) were responsible for the bts/bts2 phenotype. Bmyellow-e mRNA was strongly expressed in the trachea, testis, and integument, and expression markedly increased at the molting stages. This profile is quite similar to that of Bmyellow, a regulator of neonatal body color and body markings in Bombyx. Quantitative reverse transcription-PCR analysis showed that Bmyellow-e mRNA was heavily expressed in the integument of the head and tail in which the bts phenotype is observed. The present results suggest that Yellow-e plays a crucial role in the pigmentation process of lepidopteran larvae. PMID:19996320

  8. Transcription activator-like effector nuclease (TALEN)-mediated female-specific sterility in the silkworm, Bombyx mori

    PubMed Central

    Xu, Jun; Wang, Yueqiang; Li, Zhiqian; Ling, Lin; Zeng, Baosheng; James, Anthony A.; Tan, Anjiang; Huang, Yongping

    2015-01-01

    Engineering sex-specific sterility is critical for developing transgene-based Sterile Insect Technology. Targeted genome engineering achieved by customized ZFN, TALENs or CRIPSPR/Cas9 systems has been exploited extensively in a variety of model organisms. However, screening mutated individuals without a detectable phenotype is still challenging. In addition, genetically recessive mutations only detectable in homozygotes make the experiments time-consuming. Here we model a novel genetic system in the silkworm, Bombyx mori, that results in female-specific sterility by combining transgenesis with transcription activator–like effector nucleases (TALENs) technologies. This system induces sex-specific sterility at a high efficiency by targeting the female-specific exon of the B. mori doublesex (Bmdsx) gene, which has sex-specific splicing isoforms regulating somatic sexual development. Transgenic animals co-expressing TALEN left and right arms targeting the female-specific Bmdsx exon resulted in somatic mutations and female mutants lost fecundity due to lack of egg storage and abnormal external genitalia. The wild-type sexual dimorphism of abdominal segment was not evident in mutant females. In contrast, there were no deleterious effects in mutant male moths. The current somatic TALEN technologies provide a promising approach in future insect functional genetics, thus providing the basis for the development of attractive genetic alternatives for insect population management. PMID:25125145

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

    PubMed

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

    2016-01-01

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

  10. Vertical transmission of nucleopolyhedrovirus in the silkworm, Bombyx mori L.

    PubMed

    Khurad, A M; Mahulikar, A; Rathod, M K; Rai, M M; Kanginakudru, S; Nagaraju, J

    2004-09-01

    Nucleopolyhedrovirus (NPV) was tested for vertical transmission in the silkworm, Bombyx mori. Fifth instar larvae were exposed to four different dosages of BmNPV (830, 1300, 1800, and 2000OBs/larva) and a dosage of about 2000OBs/larva was found suitable for obtaining infected adults. Histopathological studies revealed the infection in susceptible tissues and organs initially, and at later stages of infection cycles the spermatocytes and nurse cells in the young oocytes were infected in the larval rudiments of testis and ovary, respectively. The mating of infected females with uninfected males resulted in significant reduction in fecundity (P < 0.01) and hatching of eggs (P < 0.001) due to transovarial transmission of BmNPV. Mating tests of uninfected females and infected males also confirmed venereal transmission as there was a significant reduction in hatching of eggs (P < 0.01). Further, among the F1 hybrid offspring (infected female x uninfected male) that were infected transovarially, larval progeny died at first and second instar stages, whereas those infected venereally developed acute lethal infection late and died by the end of third and fourth instar stage. PCR amplification and sequencing of 473bp of immediate early-1 (ie-1) gene of BmNPV isolated from the viral-infected parent and the F1 offspring confirmed that the viral infection is vertically transmitted to the progeny. PMID:15491594

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

    PubMed

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

    2014-10-01

    This work reports on the first study of thermally induced effect on energy transport in single filaments of silkworm (Bombyx mori) fibroin degummed mild (type 1), moderate (type 2), to strong (type 3). After heat treatment from 140 to 220°C, the thermal diffusivity of silk fibroin type 1, 2, and 3 increases up to 37.9, 20.9, and 21.5%, respectively. Our detailed scanning electron microscopy study confirms that the sample diameter change is almost negligible before and after heat treatment. Raman analysis is performed on the original and heat-treated (at 147°C) samples. After heat treatment at 147°C, the Raman peaks at 1081, 1230, and 1665 cm(-1) become stronger and narrower, indicating structural transformation from amorphous to crystalline. A structure model composed of amorphous, crystalline, and laterally ordered regions is proposed to explain the structural change by heat treatment. Owing to the close packing of more adjacent laterally ordered regions, the number and size of the crystalline regions of Bombyx mori silk fibroin increase by heat treatment. This structure change gives the observed significant thermal diffusivity increase by heat treatment. PMID:24723331

  12. Retrotransposon "Qian" mediated segmental duplication in silkworm, Bombyx mori.

    PubMed

    Xu, Yunmin; Jiang, Ning; Zou, Ziliang; Tu, Zhijian; Chen, Anli; Zhao, Qiaoling; Xiang, Zhonghuai; He, Ningjia

    2014-03-01

    Transposable elements constitute a large fraction of the eukaryotic genomes. They have the potential to alter genome structure and play a major role in genome evolution. Here, we report a segmental duplication mediated by a novel long terminal repeat (LTR) retrotransposon as the cause of an egg-shell recessive lethal mutant (l-em mutant) in silkworm (Bombyx mori). The segmental duplication resulted in the duplication of six genes and the disruption of two genes. Disruption of BmEP80 (B. mori egg protein 80), a gene encoding a major egg-shell structure protein, is likely responsible for the lethal water-loss phenotype in the l-em/l-em mutant. Our data revealed that BmEP80 is present in the inner egg-shell layer and plays important roles in resistance to water efflux form eggs. A novel LTR retrotransposon (named as "Qian") was identified and the model for the Qian-mediated chromosomal segmental duplication was proposed. Detail biochemical and genomic analyses on the l-em mutant offer an opportunity to demonstrate that an LTR retrotransposon could trigger duplication of a chromosomal segment (∼96.3 kb) and confer novel phenotype. PMID:24462715

  13. Bone regeneration by polyhedral microcrystals from silkworm virus

    PubMed Central

    Matsumoto, Goichi; Ueda, Takayo; Shimoyama, Junko; Ijiri, Hiroshi; Omi, Yasushi; Yube, Hisato; Sugita, Yoshihiko; Kubo, Katsutoshi; Maeda, Hatsuhiko; Kinoshita, Yukihiko; Arias, Duverney Gaviria; Shimabukuro, Junji; Kotani, Eiji; Kawamata, Shin; Mori, Hajime

    2012-01-01

    Bombyx mori cypovirus is a major pathogen which causes significant losses in silkworm cocoon harvests because the virus particles are embedded in micrometer-sized protein crystals called polyhedra and can remain infectious in harsh environmental conditions for years. But the remarkable stability of polyhedra can be applied on slow-release carriers of cytokines for tissue engineering. Here we show the complete healing in critical-sized bone defects by bone morphogenetic protein-2 (BMP-2) encapsulated polyhedra. Although absorbable collagen sponge (ACS) safely and effectively delivers recombinant human BMP-2 (rhBMP-2) into healing tissue, the current therapeutic regimens release rhBMP-2 at an initially high rate after which the rate declines rapidly. ACS impregnated with BMP-2 polyhedra had enough osteogenic activity to promote complete healing in critical-sized bone defects, but ACS with a high dose of rhBMP-2 showed incomplete bone healing, indicating that polyhedral microcrystals containing BMP-2 promise to advance the state of the art of bone healing. PMID:23226833

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

    PubMed Central

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

    2016-01-01

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

  15. [Electrophoresis with the mulberry leave extract and officinal peat for the rehabilitative treatment of chronic non-calculous cholecystitis].

    PubMed

    Grigorian, E G; Malerian, D S; Seropian, N S; Sarkisian, R A; Iskandarian, V A; Khachatrian, K K; Avetisian, K G

    2008-01-01

    Electrophoresis using a 5% aqueous extract of mulberry leaves and a 5% aqueous extract of Tavish peat at the right hypochondrium of patients included in the study had beneficial effect on the clinical picture of chronic non-calculous cholecystitis. Specifically, the state of the hepatobiliary system improved judging by the results of staged chromatic duodenal probing, ultrasonographic examination of biliary ducts, and bioresonance diagnostics. Also, positive changes were recorded in energy metabolism. It is recommended that electrophoresis using an aqueous mulberry leaf extract be prescribed to patients with a short duration of the disease and hyperkinetic biliary ducts and intestines. In case of an inflammatory process in these organs or hypomotor dyskinesia of biliary ducts, good results can be obtained using electrophoresis with Tavish peat extracts having a wide spectrum of action. They are indicated for the rehabilitative treatment of digestive organ pathology. PMID:19069798

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

    PubMed

    Zhu, Lin; Zhang, Yu-Qing

    2016-04-01

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

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

    PubMed

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

    2016-01-01

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

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

    PubMed Central

    Son, Mina; Lee, June Yong

    2016-01-01

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

  19. Analysis of phytoplasma-responsive sRNAs provide insight into the pathogenic mechanisms of mulberry yellow dwarf disease

    NASA Astrophysics Data System (ADS)

    Gai, Ying-Ping; Li, Yi-Qun; Guo, Fang-Yue; Yuan, Chuan-Zhong; Mo, Yao-Yao; Zhang, Hua-Liang; Wang, Hong; Ji, Xian-Ling

    2014-06-01

    The yellow dwarf disease associated with phytoplasmas is one of the most devastating diseases of mulberry and the pathogenesis involved in the disease is poorly understood. To analyze the molecular mechanisms mediating gene expression in mulberry-phytoplasma interaction, the comprehensive sRNA changes of mulberry leaf in response to phytoplasma-infection were examined. A total of 164 conserved miRNAs and 23 novel miRNAs were identified, and 62 conserved miRNAs and 13 novel miRNAs were found to be involved in the response to phytoplasma-infection. Meanwhile, target genes of the responsive miRNAs were identified by sequencing of the degradome library. In addition, the endogenous siRNAs were sequenced, and their expression profiles were characterized. Interestingly, we found that phytoplasma infection induced the accumulation of mul-miR393-5p which was resulted from the increased transcription of MulMIR393A, and mul-miR393-5p most likely initiate the biogenesis of siRNAs from TIR1 transcript. Based on the results, we can conclude that phytoplasma-responsive sRNAs modulate multiple hormone pathways and play crucial roles in the regulation of development and metabolism. These responsive sRNAs may work cooperatively in the response to phytoplasma-infection and be responsible for some symptoms in the infected plants.

  20. Effect of High Pressure Homogenization and Dimethyl Dicarbonate (DMDC) on Microbial and Physicochemical Qualities of Mulberry Juice.

    PubMed

    Yu, Yuanshan; Wu, Jijun; Xu, Yujuan; Xiao, Gengsheng; Zou, Bo

    2016-03-01

    In this study, the effect of high pressure homogenization (HPH) and dimethyl dicarbonate (DMDC) on microbial and nutrient qualities of mulberry juice was evaluated. Results showed that repeated HPH passes at 200 MPa or adding DMDC at 250 mg/L significantly inactivated the indigenous microorganisms in mulberry juice (P < 0.05), whereas some surviving microorganisms recovered to grow during storage of 4 °C. The combined treatment with 3 passes of HPH and 250 mg/L of DMDC (HPH-DMDC) decreased the population of surviving indigenous microorganisms to the level attained by heat treatment at 95 °C for 1 min (HT) with no significant increase (P > 0.05) in the population of microorganisms during subsequent storage at 4 °C. Moreover, no significant changes (P > 0.05) in the physical attributes, including pH, TSS ((o) Brix), L*, a*, and b* values were observed in the samples treated by the HPH-DMDC or by HT. Compared with HT, HPH-DMDC treatment resulted in a higher degree of retention in total phenolics, and α-glucosidase inhibitory activity, although the treatment led to higher losses in cyanidin 3-glucoside, cyanidin 3-rutinoside, and antioxidant capacity. Overall, HPH-DMDC treatment can be a useful alternative to conventional thermal pasteurization of mulberry juice, considering its ability to inactive, and inhibit indigenous microorganisms. PMID:26764561

  1. Extraction, purification and anti-fatigue activity of γ-aminobutyric acid from mulberry (Morus alba L.) leaves.

    PubMed

    Chen, Hengwen; He, Xuanhui; Liu, Yan; Li, Jun; He, Qingyong; Zhang, Cuiying; Wei, Benjun; Zhang, Ye; Wang, Jie

    2016-01-01

    Mulberry (Morus alba L.) is a tree species of Moraceae widely distributed in Southern China. In the present study, the white crystal of γ-aminobutyric acid (GABA) was purified from mulberry leaves, and its bioactivity was also investigated. The main results were as follows: first, the crude GABA was extracted from mulberry leaves by using biochemical methods. Then, the crude was purified by chromatography over an S-8 macroporous resin, Sephadex G-10, and 732 cation exchange resin to yield a white crystal. Lavage administration and exposure of GABA to male NIH mice showed no adverse effects on their growth and development. In an endurance capacity test, the average loaded-swimming time of medium dose was 111.60% longer than the control (P < 0.01). Further investigations showed that relative to that of model control, the respective blood lactate (BL) concentrations of low- and medium-dose were 28.52% and 28.81% lower (P < 0.05), whereas the levels of blood urea nitrogen (BUN) were 36.83% and 40.54% lower (P < 0.05), and that of liver glycogen (LG) levels were 12.81% and 17.22% lower (P < 0.05). The results indicated that GABA has an advantage over taurine of anti-fatigue effect. These findings were indicative of the anti-fatigue activity of GABA. PMID:26743028

  2. Mulberry anthocyanins inhibit oleic acid induced lipid accumulation by reduction of lipogenesis and promotion of hepatic lipid clearance.

    PubMed

    Chang, Jia-Jen; Hsu, Man-Jung; Huang, Hui-Pei; Chung, Dai-Jung; Chang, Yun-Ching; Wang, Chau-Jong

    2013-06-26

    Mulberry (Morus alba L.) has been considered to possess different benefits such as protecting liver; improving fever, urine excretion disorder, hypertension, and diabetic syndrome; and preventing cardiovascular diseases. Recently, mounting evidence has shown that mulberry anthocyanin extract (MAE) is beneficial to hyperlipidemia; however, the mechanisms remain unclear. The present study was aimed to investigate the protective effects of MAE on hepatocyte cultured with high fatty acid and the underlying mechanisms. By using human hepatoma cell HepG2 as cell model, the results showed that MAE suppressed fatty acid synthesis and enhanced fatty acid oxidation, contributing to amelioration of lipid accumulation induced by oleic acid (OA). Moreover, MAE also inhibited acetyl coenzyme A carboxylase (ACC) activities by stimulating adenosine monophosphate-activated protein kinase (AMPK). MAE attenuated the expression of sterol regulatory element-binding protein-1 (SREBP-1) and its target molecules, such as fatty acid synthase (FAS). Similar results were also found in the expressions of enzymes involved in triglyceride and cholesterol biosyntheses including glycerol-3-phosphate acyltransferase (GPAT), 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGCoR), adipocyte-specific fatty acid binding protein (A-FABP), and SREBP-2. In contrast, the lipolytic enzyme expressions of peroxisome proliferator activated receptor α (PPARα) and carnitinepalmitol- transferase-1 (CPT1) were increased. This study suggests the hypolipidemic effects of MAE occur via phosphorylation of AMPK and inhibition of lipid biosynthesis and stimulation of lipolysis. Therefore, the mulberry anthocyanins may actively prevent nonalcoholic fatty liver disease. PMID:23731091

  3. Extraction, purification and anti-fatigue activity of γ-aminobutyric acid from mulberry (Morus alba L.) leaves

    NASA Astrophysics Data System (ADS)

    Chen, Hengwen; He, Xuanhui; Liu, Yan; Li, Jun; He, Qingyong; Zhang, Cuiying; Wei, Benjun; Zhang, Ye; Wang, Jie

    2016-01-01

    Mulberry (Morus alba L.) is a tree species of Moraceae widely distributed in Southern China. In the present study, the white crystal of γ-aminobutyric acid (GABA) was purified from mulberry leaves, and its bioactivity was also investigated. The main results were as follows: first, the crude GABA was extracted from mulberry leaves by using biochemical methods. Then, the crude was purified by chromatography over an S-8 macroporous resin, Sephadex G-10, and 732 cation exchange resin to yield a white crystal. Lavage administration and exposure of GABA to male NIH mice showed no adverse effects on their growth and development. In an endurance capacity test, the average loaded-swimming time of medium dose was 111.60% longer than the control (P < 0.01). Further investigations showed that relative to that of model control, the respective blood lactate (BL) concentrations of low- and medium-dose were 28.52% and 28.81% lower (P < 0.05), whereas the levels of blood urea nitrogen (BUN) were 36.83% and 40.54% lower (P < 0.05), and that of liver glycogen (LG) levels were 12.81% and 17.22% lower (P < 0.05). The results indicated that GABA has an advantage over taurine of anti-fatigue effect. These findings were indicative of the anti-fatigue activity of GABA.

  4. Anti-Inflammatory and Antinociceptive Properties of Flavonoids from the Fruits of Black Mulberry (Morus nigra L.)

    PubMed Central

    Chen, Hu; Pu, Junsong; Liu, Dan; Yu, Wansha; Shao, Yunying; Yang, Guangwei; Xiang, Zhonghuai; He, Ningjia

    2016-01-01

    We analyzed the anti-inflammatory and antinociceptive activities of total flavonoids (TF) found in black mulberry fruits. The TF content was 20.9 mg/g (dry weight). Two anthocyanins, cyanidin-3-O-glucoside (8.3 mg/g) and cyanidin-3-O-rutinoside (2.9 mg/g), were identified in the fruits by UPLC. The TF of black mulberry fruits had significant reducing power and radical (OH-, O2.−, DPPH and ABTS) scavenging activities that was demonstrated in a dose-response curve. The TF had inhibitory activities on xylene-induced ear edema and carrageenan-induced paw edema in mice. In addition, TF had antinociceptive activities in the two nociceptive phases of formalin test. We used ELISA to detect the pro-inflammatory cytokines IL-1β, TNF-α, IFN-γ, and NO in the serum of mice. These cytokines were significantly inhibited or scavenged by TF (50 and 100 mg/kg). The results demonstrated that TF of black mulberry possess anti-inflammatory and analgesic effects that might correlate to its antioxidant activities and inhibition of pro-inflammatory cytokines. PMID:27046026

  5. Inhibitive Effects of Mulberry Leaf-Related Extracts on Cell Adhesion and Inflammatory Response in Human Aortic Endothelial Cells

    PubMed Central

    Chao, P.-Y.; Lin, K.-H.; Chiu, C.-C.; Yang, Y.-Y.; Huang, M.-Y.; Yang, C.-M.

    2013-01-01

    Effects of mulberry leaf-related extracts (MLREs) on hydrogen peroxide-induced DNA damage in human lymphocytes and on inflammatory signaling pathways in human aortic endothelial cells (HAECs) were studied. The tested MLREs were rich in flavonols, especially bombyx faces tea (BT) in quercetin and kaempferol. Polyphenols, flavonoids, and anthocyanidin also abounded in BT. The best trolox equivalent antioxidant capacity (TEAC) was generated from the acidic methanolic extracts of BT. Acidic methanolic and water extracts of mulberry leaf tea (MT), mulberry leaf (M), and BT significantly inhibited DNA oxidative damage to lymphocytes based on the comet assay as compared to the H2O2-treated group. TNF-α-induced monocyte-endothelial cell adhesion was significantly suppressed by MLREs. Additionally, nuclear factor kappa B (NF-κB) expression was significantly reduced by BT and MT. Significant reductions were also observed in both NF-κB and activator protein (AP)-1 DNA binding by MLREs. Significant increases in peroxisome proliferator-activated receptor (PPAR) α and γ DNA binding by MLREs were also detected in M and MT extracts, but no evidence for PPAR α DNA binding in 50 μg/mL MT extract was found. Apparently, MLREs can provide distinct cytoprotective mechanisms that may contribute to its putative beneficial effects on suppressing endothelial responses to cytokines during inflammation. PMID:24371453

  6. Effects of Freeze-dried Mulberry on Antioxidant Activities and Fermented Characteristics of Yogurt during Refrigerated Storage

    PubMed Central

    2015-01-01

    This study investigated the effect of added freeze-dried mulberry fruit juice (FDMJ) (1, 3 and 5%) on the antioxidant activity and fermented characteristic of yogurt during refrigerated storage. A decrease in pH of yogurt and increase in acidity was observed during fermentation. The yogurts with FDMJ exhibited faster rate of pH reduction than control. Initial lactic acid bacteria count of yogurt was 6.49-6.94 Log CFU/g and increased above 9 Log CFU/g in control and 1% in FDMJ yogurt for 24 h. The total polyphenol and anthocyanin content of FDMJ yogurt was higher than that of control due to the presence of phytochemical contents in mulberry. Moreover, antioxidant activity such as DPPH and reducing power was highest 5% FDMJ yogurt. During cold storage, pH decreased or remained constant in all yogurts with values ranging from 4.08 to 4.78 units. In sensory evaluation, the score of 1% FDMJ yogurt was ranked higher when compared with other yogurts. It is proposed that mulberry fruit juice powder can be used to improve sensory evaluation and enhance functionality of yogurt. PMID:26877641

  7. Extraction, purification and anti-fatigue activity of γ-aminobutyric acid from mulberry (Morus alba L.) leaves

    PubMed Central

    Chen, Hengwen; He, Xuanhui; Liu, Yan; Li, Jun; He, Qingyong; Zhang, Cuiying; Wei, Benjun; Zhang, Ye; Wang, Jie

    2016-01-01

    Mulberry (Morus alba L.) is a tree species of Moraceae widely distributed in Southern China. In the present study, the white crystal of γ-aminobutyric acid (GABA) was purified from mulberry leaves, and its bioactivity was also investigated. The main results were as follows: first, the crude GABA was extracted from mulberry leaves by using biochemical methods. Then, the crude was purified by chromatography over an S-8 macroporous resin, Sephadex G-10, and 732 cation exchange resin to yield a white crystal. Lavage administration and exposure of GABA to male NIH mice showed no adverse effects on their growth and development. In an endurance capacity test, the average loaded-swimming time of medium dose was 111.60% longer than the control (P < 0.01). Further investigations showed that relative to that of model control, the respective blood lactate (BL) concentrations of low- and medium-dose were 28.52% and 28.81% lower (P < 0.05), whereas the levels of blood urea nitrogen (BUN) were 36.83% and 40.54% lower (P < 0.05), and that of liver glycogen (LG) levels were 12.81% and 17.22% lower (P < 0.05). The results indicated that GABA has an advantage over taurine of anti-fatigue effect. These findings were indicative of the anti-fatigue activity of GABA. PMID:26743028

  8. Anti-Inflammatory and Antinociceptive Properties of Flavonoids from the Fruits of Black Mulberry (Morus nigra L.).

    PubMed

    Chen, Hu; Pu, Junsong; Liu, Dan; Yu, Wansha; Shao, Yunying; Yang, Guangwei; Xiang, Zhonghuai; He, Ningjia

    2016-01-01

    We analyzed the anti-inflammatory and antinociceptive activities of total flavonoids (TF) found in black mulberry fruits. The TF content was 20.9 mg/g (dry weight). Two anthocyanins, cyanidin-3-O-glucoside (8.3 mg/g) and cyanidin-3-O-rutinoside (2.9 mg/g), were identified in the fruits by UPLC. The TF of black mulberry fruits had significant reducing power and radical (OH-,[Formula: see text], DPPH and ABTS) scavenging activities that was demonstrated in a dose-response curve. The TF had inhibitory activities on xylene-induced ear edema and carrageenan-induced paw edema in mice. In addition, TF had antinociceptive activities in the two nociceptive phases of formalin test. We used ELISA to detect the pro-inflammatory cytokines IL-1β, TNF-α, IFN-γ, and NO in the serum of mice. These cytokines were significantly inhibited or scavenged by TF (50 and 100 mg/kg). The results demonstrated that TF of black mulberry possess anti-inflammatory and analgesic effects that might correlate to its antioxidant activities and inhibition of pro-inflammatory cytokines. PMID:27046026

  9. Characterization and expression profiles of MaACS and MaACO genes from mulberry (Morus alba L.)*

    PubMed Central

    Liu, Chang-ying; Lü, Rui-hua; Li, Jun; Zhao, Ai-chun; Wang, Xi-ling; Diane, Umuhoza; Wang, Xiao-hong; Wang, Chuan-hong; Yu, Ya-sheng; Han, Shu-mei; Lu, Cheng; Yu, Mao-de

    2014-01-01

    1-Aminocyclopropane-1-carboxylic acid synthase (ACS) and 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) are encoded by multigene families and are involved in fruit ripening by catalyzing the production of ethylene throughout the development of fruit. However, there are no reports on ACS or ACO genes in mulberry, partly because of the limited molecular research background. In this study, we have obtained five ACS gene sequences and two ACO gene sequences from Morus Genome Database. Sequence alignment and phylogenetic analysis of MaACO1 and MaACO2 showed that their amino acids are conserved compared with ACO proteins from other species. MaACS1 and MaACS2 are type I, MaACS3 and MaACS4 are type II, and MaACS5 is type III, with different C-terminal sequences. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) expression analysis showed that the transcripts of MaACS genes were strongly expressed in fruit, and more weakly in other tissues. The expression of MaACO1 and MaACO2 showed different patterns in various mulberry tissues. MaACS and MaACO genes demonstrated two patterns throughout the development of mulberry fruit, and both of them were strongly up-regulated by abscisic acid (ABA) and ethephon. PMID:25001221

  10. Mulberry (桑葚子 Sang Shèn Zǐ) and its Bioactive Compounds, the Chemoprevention Effects and Molecular Mechanisms In Vitro and In Vivo

    PubMed Central

    Huang, Hui-Pei; Ou, Ting-Tsz; Wang, Chau-Jong

    2013-01-01

    Mulberry (桑葚子 sāng shèn zǐ), a traditional Chinese medicine (TCM) in Taiwan, has many bioactive substances, including polyphenol and anthocyanins compounds. Over the past decade, many scientific and medical studies have examined mulberry fruit for its antioxidation and antiinflammation effects both in vitro and in vivo. This review thus focuses on the recent advances of mulberry extracts (MEs) and their applications in the prevention and treatment of human cancer, liver disease, obesity, diabetes, and cardiovascular disease. The ME modulates several apoptotic pathways and matrix metalloproteinases (MMPs) to block cancer progression. Mulberry can increase detoxicated and antioxidant enzyme activities and regulate the lipid metabolism to treat hepatic disease resulting from alcohol consumption, high fat diet, lipopolysaccharides (LPS) and CCl4 exposure. Of the various compounds in ME, cyanidin 3-glucoside (C3G) is the most abundant, and the active compound studied in mulberry research. Herein, the antioxidant and antiinflammatory actions of C3G to improve diabetes and cardiovascular disease are also discussed. These studies provide strong evidence ME may possess the bioactivity to affect the pathogenesis of several chronic diseases. PMID:24716151

  11. Identification and characterization of Polycomb group genes in the silkworm, Bombyx mori.

    PubMed

    Li, Zhiqing; Tatsuke, Tsuneyuki; Sakashita, Kosuke; Zhu, Li; Xu, Jian; Mon, Hiroaki; Lee, Jae Man; Kusakabe, Takahiro

    2012-05-01

    Polycomb group (PcG) proteins are involved in chromatin modifications for maintaining gene repression that play important roles in the regulation of gene expression, tumorigenesis, chromosome X-inactivation, and genomic imprinting in Drosophila melanogaster, mammals, and even plants. To characterize the orthologs of PcG genes in the silkworm, Bombyx mori, 13 candidates were identified from the updated silkworm genome sequence by using the fruit fly PcG genes as queries. Comparison of the silkworm PcG proteins with those from other insect species revealed that the insect PcG proteins shared high sequence similarity. High-level expressions of all the silkworm PcG genes were maintained through day 2 to day 7 of embryogenesis, and tissue microarray data on day 3 of the fifth instar larvae showed that their expression levels were relatively low in somatic tissues, except for Enhancer of zeste (E(Z)). In addition, knockdown of each PRC2 component, such as E(Z), Extra sex combs (ESC), and Suppressor of zeste 12 (SU(Z)12), considerably decreased the global levels of H3K27me3 but not of H3K27me2. Taken together, these results suggest that insect PcG proteins are highly conserved during evolution and might play similar roles in embryogenesis. PMID:22187347

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

    NASA Astrophysics Data System (ADS)

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

    2015-11-01

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

  13. 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

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

    PubMed

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

    2015-01-01

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

  15. 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

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

    PubMed Central

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

    2015-01-01

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

  17. Identification of a midgut-specific promoter in the silkworm Bombyx mori.

    PubMed

    Jiang, Liang; Cheng, Tingcai; Dang, Yinghui; Peng, Zhengwen; Zhao, Ping; Liu, Shiping; Jin, Shengkai; Lin, Ping; Sun, Qiang; Xia, Qingyou

    2013-04-19

    The midgut is an important organ for digestion and absorption of nutrients and immune defense in the silkworm Bombyx mori. In an attempt to create a tool for midgut research, we cloned the 1080 bp P2 promoter sequence (P2P) of a highly expressed midgut-specific gene in the silkworm. The transgenic line (P2) was generated via embryo microinjection, in which the expression of EGFP was driven by P2P. There was strong green fluorescence only in the midgut of P2. RT-PCR and Western blot showed that P2P was a midgut-specific promoter with activity throughout the larval stage. A transgenic truncation experiment suggested that regions -305 to -214 and +107 to +181 were very important for P2P activity. The results of this study revealed that we have identified a midgut-specific promoter with a high level of activity in the silkworm that will aid future research and application of silkworm genes. PMID:23524268

  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. Pomegranate leaves and mulberry fruit as natural sensitizers for dye-sensitized solar cells

    SciTech Connect

    Chang, Ho; Lo, Yu-Jen

    2010-10-15

    This study employs chlorophyll extract from pomegranate leaf and anthocyanin extract from mulberry fruit as the natural dyes for a dye-sensitized solar cell (DSSC). A self-developed nanofluid synthesis system is employed to prepare TiO{sub 2} nanofluid with an average particle size of 25 nm. Electrophoresis deposition was performed to deposit TiO{sub 2} nanoparticles on the indium tin oxide (ITO) conductive glass, forming a TiO{sub 2} thin film with the thickness of 11 {mu}m. Furthermore, this TiO{sub 2} thin film was sintered at 450 C to enhance the thin film compactness. Sputtering was used to prepare counter electrode by depositing Pt thin film on FTO glass at a thickness of 20 nm. The electrodes, electrolyte (I{sub 3}{sup -}), and dyes were assembled into a cell module and illuminated by a light source simulating AM 1.5 with a light strength of 100 mW/cm{sup 2} to measure the photoelectric conversion efficiency of the prepared DSSCs. According to experimental results, the conversion efficiency of the DSSCs prepared by chlorophyll dyes from pomegranate leaf extract is 0.597%, with open-circuit voltage (V{sub OC}) of 0.56 V, short-circuit current density (J{sub SC}) of 2.05 mA/cm{sup 2}, and fill factor (FF) of 0.52. The conversion efficiency of the DSSCs prepared by anthocyanin dyes from mulberry extract is 0.548%, with V{sub OC} of 0.555 V and J{sub SC} of 1.89 mA/cm{sup 2} and FF of 0.53. The conversion efficiency is 0.722% for chlorophyll and anthocyanin as the dye mixture, with V{sub OC} of 0.53 V, J{sub SC} of 2.8 mA/cm{sup 2}, and FF of 0.49. (author)

  20. Effects of phoxim on nutrient metabolism and insulin signaling pathway in silkworm midgut.

    PubMed

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

    2016-03-01

    Silkworm (Bombyx mori) is an important economic insect. Each year, poisoning caused by phoxim pesticide leads to huge economic losses in sericulture in China. Silkworm midgut is the major organ for food digestion and nutrient absorption. In this study, we found that the activity and expression of nutrition metabolism-related enzymes were dysregulated in midgut by phoxim exposure. DGE analysis revealed that 40 nutrition metabolism-related genes were differentially expressed. qRT-PCR results indicated that the expression levels of insulin/insulin growth factor signaling (IIS) pathway genes Akt, PI3K, PI3K60, PI3K110, IRS and PDK were reduced, whereas PTEN's expression was significantly increased in the midgut at 24 h after phoxim treatment. However, the transcription levels of Akt, PI3K60, PI3K110, IRS, InR and PDK were elevated and reached the peaks at 48 h, which were 1.48-, 1.35-, 1.21-, 2.24-, 2.89-, and 1.44-fold of those of the control, respectively. At 72 h, the transcription of these genes was reduced. Akt phosphorylation level was increasing along with the growth of silkworms in the control group. However, phoxim treatment led to increased Akt phosphorylation that surged at 24 h but gradually decreased at 48 h and 72 h. The results indicated that phoxim dysregulated the expression of IIS pathway genes and induced abnormal nutrient metabolism in silkworm midgut, which may be the reason of the slow growth of silkworms. PMID:26741554

  1. Quantitative PCR for detection of Nosema bombycis in single silkworm eggs and newly hatched larvae.

    PubMed

    Fu, Zhangwuke; He, Xiangkang; Cai, Shunfeng; Liu, Han; He, Xinyi; Li, Mingqian; Lu, Xingmeng

    2016-01-01

    Pebrine disease is the only mandatory quarantine item in sericultural production due to its destructive consequences. So far, the mother moth microscopic examination method established by Pasteur (1870) remains the only detection method for screening for the causative agent Nosema bombycis (N. bombycis). Because pebrine is a horizontal and vertical transmission disease, it is better to inspect silkworm eggs and newly hatched larvae to investigate the infection rate, vertical transmission rate and spore load of the progenies. There is a rising demand for a more direct, effective and accurate detection approach in the sericultural industry. Here, we developed a molecular detection approach based on real-time quantitative PCR (qPCR) for pebrine inspection in single silkworm eggs and newly hatched larvae. Targeting the small-subunit rRNA gene of N. bombycis, this assay showed high sensitivity and reproducibility. Ten spores in a whole sample or 0.1 spore DNA (1 spore DNA represents the DNA content of one N. bombycis spore) in a reaction system was estimated as the detection limit of the isolation and real-time qPCR procedure. Silkworm egg tissues impact the detection sensitivity but are not significant in single silkworm egg detection. Of 400 samples produced by infected moths, 167 and 195 were scored positive by light microscopy and real-time qPCR analysis, respectively. With higher accuracy and the potential capability of high-throughput screening, this method is anticipated to be adaptable for pebrine inspection and surveillance in the sericultural industry. In addition, this method can be applied to ecology studies of N. bombycis-silkworm interactions due to its quantitative function. PMID:26658327

  2. 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

  3. Remobilizing deleted piggyBac vector post-integration for transgene stability in silkworm.

    PubMed

    Wang, Feng; Wang, Riyuan; Wang, Yuancheng; Xu, Hanfu; Yuan, Lin; Ding, Huan; Ma, Sanyuan; Zhou, You; Zhao, Ping; Xia, Qingyou

    2015-06-01

    Deletion of transposable elements post-genomic integration holds great promise for stability of the transgene in the host genome and has an essential role for the practical application of transgenic animals. In this study, a modified piggyBac vector that mediated deletion of the transposon sequence post-integration for transgene stability in the economically important silkworm Bombyx mori was constructed. The piggyBac vector architecture contains inversed terminal repeat sequences L1, L2 and R1, which can form L1/R1 and L2/R1 types of transposition cassettes. hsp70-PIG as the piggyBac transposase expression cassette for initial transposition, further remobilization and transgene stabilization test was transiently expressed in a helper vector or integrated into the modified vector to produce a transgenic silkworm. Shortening L2 increased the transformation frequency of L1/R1 into the silkworm genome compared to L2/R1. After the integration of L1/R1 into the genome, the remobilization of L2/R1 impaired the transposon structure and the resulting transgene linked with an impaired transposon was stable in the genome even in the presence of exogenously introduced transposase, whereas those flanked by the intact transposon were highly mobile in the genome. Our results demonstrated the feasibility of post-integration deletion of transposable elements to guarantee true transgene stabilization in silkworm. We suggest that the modified vector will be a useful resource for studies of transgenic silkworms and other piggyBac-transformed organisms. PMID:25589404

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

    PubMed

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

    2015-10-01

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

  5. Worms and Trees: An Exciting Adventure.

    ERIC Educational Resources Information Center

    El Harim, Jean Love

    1997-01-01

    Outlines procedures for using silkworms in elementary science, mathematics, history, ecology, social studies, language arts, art, and physical education. Also focuses on the silkworm's use of the mulberry tree. (DDR)

  6. Extraction optimization, characterization and antioxidant activity in vitro of polysaccharides from mulberry (Morus alba L.) leaves.

    PubMed

    Yuan, Qingxia; Xie, Yufeng; Wang, Wei; Yan, Yuhua; Ye, Hong; Jabbar, Saqib; Zeng, Xiaoxiong

    2015-09-01

    Extraction optimization, characterization and antioxidant activity in vitro of polysaccharides from mulberry leaves (MLP) were investigated in the present study. The optimal extraction conditions with an extraction yield of 10.0 ± 0.5% for MLP were determined as follows: extraction temperature 92 °C, extraction time 3.5h and ratio (v/w, mL/g) of extraction solvent (water) to raw material 34. Two purified fractions, MLP-3a and MLP-3b with molecular weights of 80.99 and 3.64 kDa, respectively, were obtained from crude MLP by chromatography of DEAE-Cellulose 52 and Sephadex G-100. Fourier transform-infrared spectroscopy revealed that crude MLP, MLP-3a and MLP-3b were acidic polysaccharides. Furthermore, crude MLP and MLP-3a had more complicated monosaccharide compositions, while MLP-3b had a relatively higher content of uronic acid. Crude MLP, MLP-3a and MLP-3b exhibited potent Fe(2+) chelating power and scavenging activities on 1,1-diphenyl-2-picrylhydrazyl, hydroxyl, superoxide and 2,2'-azinobis-(3-ethyl-benzothiazolin-6-sulfonic acid) radicals. The results suggested that MLP could be explored as natural antioxidant. PMID:26005139

  7. Odisolane, a Novel Oxolane Derivative, and Antiangiogenic Constituents from the Fruits of Mulberry (Morus alba L.).

    PubMed

    Lee, Seoung Rak; Park, Jun Yeon; Yu, Jae Sik; Lee, Sung Ok; Ryu, Ja-Young; Choi, Sang-Zin; Kang, Ki Sung; Yamabe, Noriko; Kim, Ki Hyun

    2016-05-18

    Mulberry, the fruit of Morus alba L., is known as an edible fruit and commonly used in Chinese medicines as a warming agent and as a sedative, tonic, laxative, odontalgic, expectorant, anthelmintic, and emetic. Systemic investigation of the chemical constituents of M. alba fruits led to the identification of a novel oxolane derivative, (R*)-2-((2S*,3R*)-tetrahydro-2-hydroxy-2-methylfuran-3-yl)propanoic acid (1), namely, odisolane, along with five known heterocyclic compounds (2-6). The structure of the new compound was elucidated on the basis of HR-MS, 1D and 2D NMR ((1)H-(1)H COSY, HSQC, HMBC, and NOESY) data analysis. Compound 1 has a novel skeleton that consists of 8 carbon units with an oxolane ring, which until now has never been identified in natural products. The isolated compounds were subjected to several activity tests to verify their biological function. Among them, compounds 1, 3, and 5 significantly inhibited cord formation in HUVECs. The action mechanism of compound 3, which had the strongest antiangiogenic activity, was mediated by decreasing VEGF, p-Akt, and p-ERK protein expression. These results suggest that compounds isolated from M. alba fruits might be beneficial in antiangiogenesis therapy for cancer treatment. PMID:27115720

  8. Analysis and characterisation of phytochemicals in mulberry (Morus alba L.) fruits grown in Vojvodina, North Serbia.

    PubMed

    Natić, Maja M; Dabić, Dragana Č; Papetti, Adele; Fotirić Akšić, Milica M; Ognjanov, Vladislav; Ljubojević, Mirjana; Tešić, Živoslav Lj

    2015-03-15

    In this study, the polyphenolic profile of 11 Morus alba fruits grown in the Vojvodina region was investigated. Ultra high performance liquid chromatography (UHPLC) coupled with Linear Trap Quadrupole and OrbiTrap mass analyzer, and UHPLC coupled with a diode array detector and a triple-quadrupole mass spectrometer were used for the identification and quantification of the polyphenols, respectively. A total of 14 hydroxycinnamic acid esters, 13 flavonol glycosides, and 14 anthocyanins were identified in the extracts with different distributions and contents according to the sampling. The total phenolic content ranged from 43.84 to 326.29 mg GAE/100g frozen fruit. The radical scavenging capacity (50.18-86.79%), metal chelating ability (0.21-8.15%), ferric ion reducing power (0.03-38.45 μM ascorbic acid) and superoxide anion radical scavenging activity (16.53-62.83%) were assessed. The findings indicated that mulberry polyphenolics may act as potent superoxide anion radical scavengers and reducing agents. PMID:25308652

  9. Hydrologic responses of streams to mining of the Mulberry coal reserves in eastern Kansas

    USGS Publications Warehouse

    Bevans, H.E.

    1984-01-01

    The U.S. Geological Survey investigated the hydrologic responses of streams with respect to coal-mining activities in the Mulberry coal reserves of Miami, Linn, and Bourbon Counties, eastern Kansas. Results of a low-flow water-quality reconnaissance showed that small streams draining previously coal-mined areas generally have relatively large concentrations of sulfate. Large streams in the study area have been relatively unaffected by coal mining. A comparison of two small drainage basins showed that the basin affected by an active strip mine had less high flow and more low flow because of the regulating effects of sediment ponds. Effluent pumped from the strip mine increased the load of sulfate by 244 percent, the load of dissolved solids by 41 percent, and occasionally transported relatively large concentrations of iron, lead, manganese, and zinc to the receiving stream. Accelerated erosion caused by the exposure and disturbance of soil during clearing and excavation increased the sediment load of the receiving stream by 25 percent even though sediment ponds were installed. (USGS)

  10. Physical and hydrologic environments of the Mulberry coal reserves in eastern Kansas

    USGS Publications Warehouse

    Kenny, J.F.; Bevans, H.E.; Diaz, A.M.

    1982-01-01

    Strippable reserves of Mulberry coal underlie an area of approximately 300 square miles of Miami, Linn, and Bourbon Counties of eastern Kansas. Although subject to State reclamation law, current and projected strip mining of this relatively thin coal seam could alter and hydrologic environment of the study area. Drained by the Marais des Cygnes and Little Osage Rivers and their tributaries, this area is characterized by low relief and moderately impermeable soils. Streamflows are poorly sustained by ground-water discharge and fluctuate widely due to climatic extremes and usage of surface-water supplies. Because ground-water supplies are generally unreliable in quantity and quality, surface water is used to meet most water requirements in the study area. Primary used of surface waters are for domestic supplies, maintenance of wildlife and recreational areas, and cooling needs at LaCygne Power Plant. The prevailing chemical type of the natural streamflow is calcium bicarbonate, with concentrations of dissolved solids generally less than 500 milligrams per liter and pH near neutral. Additional streamflow and water-quality data are needed to evaluate the premining characteristics of and the anticipated changes in the hydrologic environment as strip mining proceeds within the study area. A network of data-collection stations and a sampling scheme have been established to acquire this additional information. (USGS)

  11. Hydrologic data for a subsurface waste-injection site at Mulberry, Florida; 1972-77

    USGS Publications Warehouse

    Wilson, William Edward; Parsons, David C.; Spechler, R.M.

    1979-01-01

    Since October 1972, industrial liquid waste has been injected into a brine aquifer of limestone and dolomite in Mulberry, FL., at a depth of more than 4,000 feet below land surface. During 1977, the injection rate was about 8.8 million gallons per month. To determine what effect the injected waste has on the ground-water body, water levels have been measured and water samples collected from two monitor wells that tap different permeable zones above the injection zone, and from a satellite monitor well that taps the injection zone. The monitor wells are in the annulus of the injection well, and the satellite monitor well is 2,291 feet from the injection well. This report updates previous data reports and includes all hydrologic data collected by the U.S. Geological Survey during 1972-77. Included is a table of well-construction data, a graph showing the volume of waste injected each month, and hydrographs of the annulus monitor wells and the satellite monitor well. (Woodard-USGS)

  12. Two chitinase-like proteins abundantly accumulated in latex of mulberry show insecticidal activity

    PubMed Central

    2010-01-01

    Background Plant latex is the cytoplasm of highly specialized cells known as laticifers, and is thought to have a critical role in defense against herbivorous insects. Proteins abundantly accumulated in latex might therefore be involved in the defense system. Results We purified latex abundant protein a and b (LA-a and LA-b) from mulberry (Morus sp.) and analyzed their properties. LA-a and LA-b have molecular masses of approximately 50 and 46 kDa, respectively, and are abundant in the soluble fraction of latex. Western blotting analysis suggested that they share sequence similarity with each other. The sequences of LA-a and LA-b, as determined by Edman degradation, showed chitin-binding domains of plant chitinases at the N termini. These proteins showed small but significant chitinase and chitosanase activities. Lectin RCA120 indicated that, unlike common plant chitinases, LA-a and LA-b are glycosylated. LA-a and LA-b showed insecticidal activities when fed to larvae of the model insect Drosophila melanogaster. Conclusions Our results suggest that the two LA proteins have a crucial role in defense against herbivorous insects, possibly by hydrolyzing their chitin. PMID:20109180

  13. Proteome analysis of silkworm, Bombyx mori, larval gonads: characterization of proteins involved in sexual dimorphism and gametogenesis.

    PubMed

    Chen, Jin-e; Li, Jian-ying; You, Zheng-ying; Liu, Li-li; Liang, Jian-she; Ma, Ying-ying; Chen, Ming; Zhang, Hua-rong; Jiang, Zhen-dong; Zhong, Bo-xiong

    2013-06-01

    Sexual dimorphism is initialed by the components of the sex determination pathway and is most evident in gonads and germ cells. Although striking dimorphic expressions have been detected at the transcriptional level between the silkworm larval testis and the ovary, the sex-dimorphic expressions at the protein level have not yet been well characterized. The proteome of silkworm larval gonads was investigated using a shotgun-based identification. A total of 286 and 205 nonredundant proteins were identified from the silkworm testis and ovary, respectively, with a false discovery rate (FDR) lower than 1%. Only 40 and 16 proteins were previously identified, and 246 and 189 proteins were newly identified in the silkworm testis and the ovary, respectively. The gametogenesis mechanism of silkworm was demonstrated using the protein expression profile and bioinformatics analysis. Cellular retinoic acid binding protein (CRABP) showed to be highly abundant in testis, while tubulins were abundant in ovary. Several homologies of Drosophila essential proteins for gametogenesis were identified in silkworm, such as male meiotic arrest gene product ALY and VISMAY in testis, and maternal mRNA localization protein exuperantia and SQUID in ovary. The gene ontology (GO) annotation and pathway analysis provide system-level insights into the sexual dimorphism and gametogenesis. PMID:23607740

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

    PubMed

    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

  15. 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

  16. Ecdysone Titer Determined by 3DE-3β-Reductase Enhances the Immune Response in the Silkworm.

    PubMed

    Sun, Wei; Shen, Yi-Hong; Zhou, Liang-Xiao; Zhang, Ze

    2016-02-15

    Although recent studies have demonstrated that 20-hydroxyecdysone (20E), one of the two most important hormones for development, could promote the insect innate immune response, how insects regulate 20E titer to affect the immunity after suffering pathogen attack remains unknown. In this study, to our knowledge, we first found that 20E titer was significantly elevated after bacterial infection in the domesticated silkworm, Bombyx mori. Furthermore, the elevated 20E enhanced the silkworm innate immune system against invading bacteria via ecdysone receptor. During immune response, the expression of the silkworm 3-dehydroecdysone-3β-reductase (3DE-3β-reductase) that converts 3DE released from prothoracic glands into ecdysone was induced. RNA interference experiments suggested that 3DE-3β-reductase is essential to upregulate the 20E titer after bacterial infection. The rescue experiments showed that injection with the recombinant 3DE-3β-reductase protein can significantly elevate the 20E concentration and modulate the expressions of the silkworm immune-related genes. Taken together, 20E titer determined by 3DE-3β-reductase enhances the silkworm defense against the bacterial infection. Thus, our findings reveal an important role of the 20E synthesis pathway from 3DE in enhancing the silkworm immune response and have profound implications for the understanding of interaction mechanisms between insect hormone and immunity. PMID:26773159

  17. Microarray Analysis of the Juvenile Hormone Response in Larval Integument of the Silkworm, Bombyx mori

    PubMed Central

    Cheng, Daojun; Peng, Jian; Meng, Meng; Wei, Ling; Kang, Lixia; Qian, Wenliang; Xia, Qingyou

    2014-01-01

    Juvenile hormone (JH) coordinates with 20-hydroxyecdysone (20E) to regulate larval growth and molting in insects. However, little is known about how this cooperative control is achieved during larval stages. Here, we induced silkworm superlarvae by applying the JH analogue (JHA) methoprene and used a microarray approach to survey the mRNA expression changes in response to JHA in the silkworm integument. We found that JHA application significantly increased the expression levels of most genes involved in basic metabolic processes and protein processing and decreased the expression of genes associated with oxidative phosphorylation in the integument. Several key genes involved in the pathways of insulin/insulin-like growth factor signaling (IIS) and 20E signaling were also upregulated after JHA application. Taken together, we suggest that JH may mediate the nutrient-dependent IIS pathway by regulating various metabolic pathways and further modulate 20E signaling. PMID:24809046

  18. Production of small antibacterial peptides using silkworm-baculovirus protein expression system.

    PubMed

    Fukushima, Mai; Iiyama, Kazuhiro; Yamashita, Jun; Furue, Masutaka; Tsuji, Gaku; Imanishi, Shigeo; Mon, Hiroaki; Lee, Jae Man; Kusakabe, Takahiro

    2013-01-01

    The recombinant proteins with strong antimicrobial activity are known to be very difficult to express using bacterial expression system. Here, human β-defensin (DEFB) 1, DEFB2, and DEFB3 were successfully produced using a silkworm-baculovirus protein expression system. We have generated four baculoviruses for each DEFB protein to compare the effect of different peptide tags in secretion into silkworm larval hemolymph. Interestingly, the best performing peptide tags for the secretion were different among DEFBs: C-terminal GST-H8 tag for DEFB1, N-terminal H8 tag for DEFB2, and C-terminal H8 tag for DEFB3, respectively. In addition, the colony count assay demonstrated that the recombinant DEFB2 s showed antimicrobial activities against Escherichia coli, Pseudomonas aeruginosa, and Paenibacillus thiaminolyticus. PMID:23742088

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

    PubMed Central

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

    2013-01-01

    Malpighian tubules (MTs) are highly specific organs of arthropods (Insecta, Myriapoda and Arachnida) for excretion and osmoregulation. In order to highlight the important genes and pathways involved in multi-functions of MTs, we performed a systematic proteomic analysis of silkworm MTs in the present work. Totally, 1,367 proteins were identified by one-dimensional gel electrophoresis coupled with liquid chromatography-tandem mass spectrometry, and as well as by Trans Proteomic Pipeline (TPP) and Absolute protein expression (APEX) analyses. Forty-one proteins were further identified by two-dimensional gel electrophoresis. Some proteins were revealed to be significantly associated with various metabolic processes, organic solute transport, detoxification and innate immunity. Our results might lay a good foundation for future functional studies of MTs in silkworm and other lepidoptera. PMID:24098719

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

    PubMed

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

    2016-02-15

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

  1. 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

  2. HPLC-MSn identification and quantification of flavonol glycosides in 28 wild and cultivated berry species.

    PubMed

    Mikulic-Petkovsek, Maja; Slatnar, Ana; Stampar, Franci; Veberic, Robert

    2012-12-15

    Berries and red fruits are rich dietary sources of polyphenols with reported health benefits. More than 50 different flavonols (glycosides of quercetin, myricetin, kaempferol, isorhamnetin, syringetin and laricitrin) have been detected and quantified with HPLC-MS(n) in fruits of blueberry, bilberry, cranberry, lingonberry, eastern shadbush, Japanese wineberry, black mulberry, chokeberry, red, black and white currants, jostaberry, red and white gooseberry, hardy kiwifruit, goji berry, rowan, dog rose, Chinese and midland hawthorn, wild and cultivated species of blackberry, raspberry, strawberry and elderberry. The phenolic constituents and contents varied considerably among the analyzed berry species. Elderberry contained the highest amount of total flavonols (450-568 mgkg(-1) FW), followed by berry species, containing more than 200 mgkg(-1) FW of total: chokeberry (267mgkg(-1)), eastern shadbush (261 mgkg(-1)), wild grown blackberry (260 mgkg(-1)), rowanberry (232 mgkg(-1)), american cranberry (213 mgkg(-1)) and blackcurrants (204 mgkg(-1)). Strawberry (10.5 mgkg(-1)) and white currants (4.5 mgkg(-1)) contained the lowest amount of total flavonols. Quercetins represent the highest percentage (46-100%) among flavonols in most analyzed berries. In wild strawberry and gooseberry the prevailing flavonols belong to the group of isorhamnetins (50-62%) and kaempferols, which represent the major part of flavonols in currants (49-66%). Myricetin glycosides could only be detected in chokeberry, rowanberry and species from the Grossulariaceae, and Adoxaceae family and Vaccinium genus. Wild strawberry and blackberry contained from 3- to 5-fold higher total flavonols than the cultivated one. PMID:22980782

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

    PubMed

    Onaga, Lisa

    2010-01-01

    Japanese agricultural scientist Toyama Kametaro's report about the Mendelian inheritance of silkworm cocoon color in Studies on the Hybridology of Insects (1906) spurred changes in Japanese silk production and thrust Toyama and his work into a scholarly exchange with American entomologist Vernon Kellogg. Toyama's work, based on research conducted in Japan and Siam, came under international scrutiny at a time when analyses of inheritance flourished after the "rediscovery" of Mendel's laws of heredity in 1900. The hybrid silkworm studies in Asia attracted the attention of Kellogg, who was concerned with how experimental biology would be used to study the causes of natural selection. He challenged Toyama's conclusions that Mendelism alone could explain the inheritance patterns of silkworm characters such as cocoon color because they had been subject to hundreds of years of artificial selection, or breeding. This examination of the intersection of Japanese sericulture and American entomology probes how practical differences in scientific interests, societal responsibilities, and silkworm materiality were negotiated throughout the processes of legitimating Mendelian genetics on opposite sides of the Pacific. The ways in which Toyama and Kellogg assigned importance to certain silkworm properties show how conflicting intellectual orientations arose in studies of the same organism. Contestation about Mendelism took place not just on a theoretical level, but the debate was fashioned through each scientist's rationale about the categorization of silkworm breeds and races and what counted as "natural". This further mediated the acceptability of the silkworm not as an experimental organism, but as an appropriately "natural" insect with which to demonstrate laws of inheritance. All these shed light on the challenges that came along with the use of agricultural animals to convincingly articulate new biological principles. PMID:20665229

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

    PubMed

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

    2015-04-01

    Dopamine is a precursor for melanin synthesis. Arylalkylamine N-acetyltransferase (AANAT) is involved in the melatonin formation in insects because it could catalyze the transformation from dopamine to dopamine-N-acetyldopamine. In this study, we identified a new AANAT gene in the silkworm (Bombyx mori) and assessed its role in the silkworm. The cDNA of this gene encodes 233 amino acids that shares 57 % amino acid identity with the Bm-iAANAT protein. We thus refer to this gene as Bm-iAANAT2. To investigate the role of Bm-iAANAT2, we constructed a transgenic interference system using a 3xp3 promoter to suppress the expression of Bm-iAANAT2 in the silkworm. We observed that melanin deposition occurs in the head and integument in transgenic lines. To verify the melanism pattern, dopamine content and the enzyme activity of AANAT were determined by high-performance liquid chromatography (HPLC). We found that an increase in dopamine levels affects melanism patterns on the heads of transgenic B. mori. A reduction in the enzyme activity of AANAT leads to changes in dopamine levels. We analyzed the expression of the Bm-iAANAT2 genes by qPCR and found that the expression of Bm-iAANAT2 gene is significantly lower in transgenic lines. Our results lead us to conclude that Bm-iAANAT2 is a new arylalkylamine N-acetyltransferase gene in the silkworm and is involved in the metabolism of the dopamine to avoid the generation of melanin. PMID:25712907

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

    PubMed Central

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

    2013-01-01

    Microsporidia have attracted much attention because they infect a variety of species ranging from protists to mammals, including immunocompromised patients with AIDS or cancer. Aside from the study on Nosema ceranae, few works have focused on elucidating the mechanism in host response to microsporidia infection. Nosema bombycis is a pathogen of silkworm pébrine that causes great economic losses to the silkworm industry. Detailed understanding of the host (Bombyx mori) response to infection by N. bombycis is helpful for prevention of this disease. A genome-wide survey of the gene expression profile at 2, 4, 6 and 8 days post-infection by N. bombycis was performed and results showed that 64, 244, 1,328, 1,887 genes were induced, respectively. Up to 124 genes, which are involved in basal metabolism pathways, were modulated. Notably, B. mori genes that play a role in juvenile hormone synthesis and metabolism pathways were induced, suggesting that the host may accumulate JH as a response to infection. Interestingly, N. bombycis can inhibit the silkworm serine protease cascade melanization pathway in hemolymph, which may be due to the secretion of serpins in the microsporidia. N. bombycis also induced up-regulation of several cellular immune factors, in which CTL11 has been suggested to be involved in both spore recognition and immune signal transduction. Microarray and real-time PCR analysis indicated the activation of silkworm Toll and JAK/STAT pathways. The notable up-regulation of antimicrobial peptides, including gloverins, lebocins and moricins, strongly indicated that antimicrobial peptide defense mechanisms were triggered to resist the invasive microsporidia. An analysis of N. bombycis-specific response factors suggested their important roles in anti-microsporidia defense. Overall, this study primarily provides insight into the potential molecular mechanisms for the host-parasite interaction between B. mori and N. bombycis and may provide a foundation for further work on host-parasite interaction between insects and microsporidia. PMID:24386341

  6. Enhancing Mulberry Leaf Meal with Urea by Pelleting to Improve Rumen Fermentation in Cattle

    PubMed Central

    Tan, N. D.; Wanapat, M.; Uriyapongson, S.; Cherdthong, A.; Pilajun, R.

    2012-01-01

    Four, ruminally fistulated crossbred (Brahman×native) beef cattle with initial body weight of 420±15 kg were randomly assigned according to a 4×4 Latin square design. The dietary treatments were mulberry leaf pellet (MUP) supplementation at 0, 200, 400 and 600 g/hd/d with rice straw fed to allow ad libitum intake. All steers were kept in individual pens and supplemented with concentrate at 5 g/kg of body weight daily. The experiment was 4 periods, and each lasted 21 d. During the first 14 d, all steers were fed their respective diets ad libitum and during the last 7 d, they were moved to metabolism crates for total urine and fecal collection. It was found that increasing MUP levels resulted in linearly increasing rice straw and total intakes (p<0.05). Ruminal temperature and pH were not significantly affected by MUP supplementation while NH3-N concentration was increased (p<0.05) and maintained at a high level (18.5 mg/dl) with supplementation of MUP at 600 g/hd/d. Similarly, viable total bacteria in the rumen and cellulolytic bacteria were enriched by MUP supplementation at 600 g/hd/d. However, the rumen microbial diversity determined with a PCR-DGGE technique showed similar methanogenic diversity between treatments and sampling times and were similar at a 69% genetic relationship as determined by a UPGMA method. Based on this study, it could be concluded that supplementation of MUP at 600 g/hd/d improved DM intake, ruminal NH3-N, and cellulolytic bacteria thus iimproving rumen ecology in beef cattle fed with rice straw. PMID:25049585

  7. Turned windrow composting of cow manure as appropriate technology for zero discharge of mulberry pulp wastewater.

    PubMed

    Jolanun, Banjarata; Kaewkam, Chompoonuch; Bauoon, Orapin; Chiemchaisri, Chart

    2014-08-01

    Turned windrow composting was investigated as appropriate technology for recycling the wastewater (excluding black liquor) from mulberry pulp and paper handicrafts. Two exterior turned windrows (1.5 m width x 1.5 m height x 2.0 m length) with dry leaves/cow manure/sawdust wet weight ratios of 60:40:0 (Pile A) and 55:40:5 (Pile B) were used for the investigation. Changes in the physical and chemical properties of the compost were examined and a phytotoxicity analysis was performed. A soil incubation test and an informal focus group discussion were also conducted. The results revealed that while both piles met the regulatory processing requirements for further reduced pathogens (>or= 55 degrees C for 15 days or longer), the operation without sawdust (Pile A) not only significantly enhanced the thermophilic temperature regime (P < 0.05) but also yielded the highest amount (1.4 m3 ton-1 pile) of wastewater elimination during the first 2 months of composting. It was found that the constant rates of degradation were 0.006 day- 1 (Pile A) and 0.003 day-1 (Pile B), and no pronounced statistically significant difference in N losses was found (P > 0.05). The germination index of two plant species in both piles varied between 126% and 230% throughout the experiment, and no pronounced differences (P > 0.05) among the samples were found. Addition of the compost significantly improved soil organic matter and pH (7-8), as well as reduced the loss of NO3-N. Local discussion groups were initiated to evaluate the cost-benefits, the potential of wastewater removal, the cooperation of community users and supporters, the compost quality and the potential compost market. PMID:24956805

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

    PubMed

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

    2015-05-01

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

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

    SciTech Connect

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

    2009-07-31

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

  10. 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

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

    PubMed

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

    2014-10-01

    As an important economic insect, Bombyx mori is also a useful model organism for lepidopteran insect. Integrins are evolutionarily conserved from sponges to humans, and play vital roles in many physiological and pathological processes. To explore their diverse functions of integrins in insect, eleven integrins including six α and five β subunits were cloned and characterized from silkworm. Our results showed that integrins from silkworm own more family members compared to other invertebrates. Among those α subunits, integrins α1, α2, and the other four subunits belong to PS1, PS2, and PS3 groups, respectively. The β subunits mainly gather in the insect βν group except the β1 subunit which belongs to the insect β group. Expression profiles demonstrated that the integrins exhibited distinct patterns, but were mainly expressed in hemocytes. α1 and β2 subunits are the predominant ones either in the embryogenesis or larva stages. Interestingly, integrins were significantly up-regulated after stimulated by 20-hydroxyecdysone (20-E) in vivo. These results indicate that integrins perform diverse functions in hemocytes of silkworm. Overall, our results provide a new insight into the functional and evolutionary features of integrins. PMID:25064490

  12. 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.

  13. 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

  14. Genetic characterisation of microsporidia infecting Indian tasar silkworm, Antheraea mylitta, using morphology and molecular tools.

    PubMed

    Hassan, Wazid; Surendra Nath, B

    2015-01-01

    The utility of inter simple sequence repeat-PCR (ISSR-PCR) assay in the genetic characterisation and elucidation of the phylogenetic relationship of different microsporidian isolates infecting tropical tasar silkworm, Antheraea mylitta Drury, is demonstrated. A total of 22 different microsporidians collected from the diseased tasar silkworms from Jharkhand state of India were analysed using morphological characters and ISSR-PCR. Observations spores under phase contrast microscope revealed oval to elongate in shape with length ranging from 3.8 μm to 5.1 μm and width from 2.6 μm to 3.3 μm. All the microsporidian isolates except MIJ-1gC showed gonadal infection and transovarial transmission in infected tasar silkworms. Fourteen out of 20 ISSR primers tested generated reproducible profiles and yielded a total of 281 fragments, of which 273 were polymorphic (97%). The degree of banding pattern was used to evaluate genetic distances and for phylogenetic analysis. The results demonstrated that ISSR analysis may be a useful and efficient tool for taxonomical grouping and phylogenetic classification of different microsporidians in general. PMID:26198429

  15. 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

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

    PubMed

    Fu, Ping; Sun, Wei; Zhang, Ze

    2016-04-10

    Acylpeptide hydrolase (APH) can catalyze the release of the N-terminal amino acid from acetylated peptides. There were many documented examples of this enzyme in various prokaryotic and eukaryotic organisms. However, knowledge about APH in insects still remains unknown. In this study, we cloned and sequenced a putative silkworm Bombyx mori APH (BmAPH) gene. The BmAPH gene encodes a protein of 710 amino acids with a predicted molecular mass of 78.5kDa. The putative BmAPH and mammal APHs share about 36% amino acid sequence identity, yet key catalytic residues are conserved (Ser566, Asp654, and His686). Expression and purification of the recombinant BmAPH in Escherichia coli showed that it has acylpeptide hydrolase activity toward the traditional substrate, Ac-Ala-pNA. Furthermore, organophosphorus (OP) insecticides, chlorpyrifos, phoxim, and malathion, significantly inhibited the activity of the APH both in vitro and in vivo. In addition, BmAPH was expressed in all tested tissues and developmental stages of the silkworm. Finally, immunohistochemistry analysis showed that BmAPH protein was localized in the basement membranes. These results suggested that BmAPH may be involved in enhancing silkworm tolerance to the OP insecticides. In a word, our results provide evidence for understanding of the biological function of APH in insects. PMID:26778207

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

    PubMed Central

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

    2015-01-01

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

  18. Xylella fastidiosa isolates from mulberry harbor a 25 kilobase pair plasmid with extensive sequence identity to a plasmid from Verminephrobacter eiseniae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A 25 kbp plasmid was present in each of four Californian isolates of Xylella fastidiosa from mulberry affected with leaf scorch disease. Fragments of each plasmid were cloned into E. coli, sequenced, and assembled into circular contigs of 25,105 bp (pXF-RIV11 and pXF-RIV16) or 24,372 bp (pXF-RIV19 a...

  19. Mulberry strains of Xylella fastidiosa contain a 25 kilobase pair plasmid with extensive sequence identity to a plasmid from Verminephrobacter eiseniae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A 25 kbp plasmid was present in each of four Californian strains of Xylella fastidiosa from mulberry affected with leaf scorch disease. Fragments of each plasmid were cloned into E. coli, sequenced, and assembled into circular contigs of 25,105 bp (pXF-RIV11 and pXF-RIV16) or 24,372 bp (pXF-RIV19 an...

  20. Plasmids of Xylella fastidiosa Mulberry-Infecting Strains Share Extensive Sequence Identity and Gene Complement with pVEIS01 From the Earthworm Symbiont Verminephrobacter Eiseniae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A ~25 kbp plasmid was present in each of four Californian strains of Xylella fastidiosa from mulberry affected with leaf scorch disease. Fragments of each plasmid were cloned into E. coli, sequenced, and assembled into circular contigs of 25,105 bp (pXF-RIV11 and pXF-RIV16) or 24,372 bp (pXF-RIV19 a...

  1. Promotion of mitotic catastrophe via activation of PTEN by paclitaxel with supplement of mulberry water extract in bladder cancer cells

    PubMed Central

    Chen, Nien-Cheng; Chyau, Charng-Cherng; Lee, Yi-Ju; Tseng, Hsien-Chun; Chou, Fen-Pi

    2016-01-01

    Paclitaxel is a mitotic inhibitor used in cancer chemotherapy. Mulberry fruit is rich in phenolic compounds and flavonoids and exhibits chemopreventive activities. In this study, mulberry water extract (MWE) was used as a supplement to synergize with the effects of paclitaxel in the treatment of the TSGH 8301 human bladder cancer cell line. Treatment with paclitaxel combined with MWE (paclitaxel/MWE) enhanced the cytotoxicity of paclitaxel and induced severe G2/M arrest, mitotic catastrophe and subsequent apoptosis, as shown by MTT assay, HE staining and flow cytometry analyses. Differences in the expression and activation of Aurora A and Plk1between cells treated with paclitaxel/MWE and paclitaxel alone suggested that the combined treatment caused a defect in the early steps of cytokinesis. Paclitaxel/MWE decreased EEA1immunofluorescence staining and increased the expression of PTEN, indicating that the regimen inhibited the formation of the recycling endosome, which is required for cytokinesis. Paclitaxel/MWE also retarded tumor growth in a TSGH 8301 xenograft model via activation of PTEN and Caspase 3. These data demonstrated a synergistic effect on the anticancer efficacy of paclitaxel through MWE supplementation by promoting mitotic catastrophe through the activation of PTEN, providing a novel and effective therapeutic option for bladder cancer treatment strategies. PMID:26838546

  2. Genome-Wide Identification and Characterization of Long Non-Coding RNAs from Mulberry (Morus notabilis) RNA-seq Data.

    PubMed

    Song, Xiaobo; Sun, Liang; Luo, Haitao; Ma, Qingguo; Zhao, Yi; Pei, Dong

    2016-01-01

    Numerous sources of evidence suggest that most of the eukaryotic genome is transcribed into protein-coding mRNAs and also into a large number of non-coding RNAs (ncRNAs). Long ncRNAs (lncRNAs), a group consisting of ncRNAs longer than 200 nucleotides, have been found to play critical roles in transcriptional, post-transcriptional, and epigenetic gene regulation across all kingdoms of life. However, lncRNAs and their regulatory roles remain poorly characterized in plants, especially in woody plants. In this paper, we used a computational approach to identify novel lncRNAs from a published RNA-seq data set and analyzed their sequences and expression patterns. In total, 1133 novel lncRNAs were identified in mulberry, and 106 of these lncRNAs displayed a predominant tissue-specific expression in the five major tissues investigated. Additionally, functional predictions revealed that tissue-specific lncRNAs adjacent to protein-coding genes might play important regulatory roles in the development of floral organ and root in mulberry. The pipeline used in this study would be useful for the identification of lncRNAs obtained from other deep sequencing data. Furthermore, the predicted lncRNAs would be beneficial towards an understanding of the variations in gene expression in plants. PMID:26938562

  3. Gamma-phase homogenization and texture in U-7.5Nb-2.5Zr (Mulberry) alloy

    NASA Astrophysics Data System (ADS)

    Lopes, Denise Adorno; Restivo, Thomaz Augusto Guisard; de Lima, Nelson Batista; Padilha, Angelo Fernando

    2014-06-01

    This work investigates the phenomena of homogenization and texture of the γ phase in U-7.5Nb-2.5Zr (Mulberry) alloy prepared by induction melting and cold-rolling. The microstructural characterization of the as-cast and homogenized alloy (heat treated in γ phase region and then quenched in water), as well as the deformed state, was performed using optical and electron microscopy techniques, hardness testing and X-ray diffraction, employing the Rietveld method. The as-cast microsegregation was qualitatively observed by optical microscopy whereas the quantitative evaluation was obtained by electronprobe micro-analysis (EPMA). The homogenization state of the structure was evaluated after heat treatment at 1000 °C in a tube furnace for 5 h. It was found that this treatment is effective in eliminate dendritic segregation in this alloy. The texture of the Mulberry alloy was studied by X-ray diffraction (XRD) in the γ-phase stabilized condition and deformed state (rolled at room temperature). The stabilized γ alloy showed moderate texture mainly on the components (0 2 3)<1 0 0> and (0 3 2)<1 0 0>. After 80% deformation, the sample showed a fiber texture (0 0 1), not commonly found in BCC metals, besides the γ fiber (1 1 1) with intermediate intensity.

  4. Characterization of polysaccharide fractions in mulberry fruit and assessment of their antioxidant and hypoglycemic activities in vitro.

    PubMed

    Chen, Chun; You, Li-Jun; Abbasi, Arshad Mehmood; Fu, Xiong; Liu, Rui Hai; Li, Chao

    2016-01-01

    The present study aimed to characterize and investigate the polysaccharide fractions of mulberry fruit, and their antioxidant and hypoglycemic activities. Mulberry fruit polysaccharides (MFP) were extracted by using hot water as the solvent and fractioned by using a DEAE-Sepharose fast-flow column. In total four eluents of water (MFP-1), 0.1 M NaCl (MFP-2), 0.2 M NaCl (MFP-3) and 0.3 M NaCl (MFP-4) were fractionated. Arabinose, galactose, glucose, xylose and mannose were the main components present in MFP-1, while MFP-2 and MFP-3 were composed of arabinose and galactose, and MFP-4 of arabinose, galactose and glucose. High-performance gel permeation chromatography (HP-GPC) analysis indicated a narrower molecular weight distribution in the MFP-1 fraction. Scanning electron microscopy (SEM) exhibited a smooth surface for MFP-1 and MFP-3 fractions, whereas MFP-2 and MFP-4 have pore openings and flocculent fibers on a smooth surface. Tertiary structure analyses indicated that none of the fractions had a triple-helical conformation. On the whole, MFP-3 and MFP-4 showed better antioxidant activity and inhibitory effects on α-amylase and α-glucosidase compared to MFP-1 and MFP-2. These results show significant potential for the antioxidant and hypoglycemic activities of MFP-3 and MFP-4 indicating the need for their further exploration as potential antidiabetic agents. PMID:26569512

  5. Use of mulberry-soybean intercropping in salt-alkali soil impacts the diversity of the soil bacterial community.

    PubMed

    Li, Xin; Sun, Minglong; Zhang, Huihui; Xu, Nan; Sun, Guangyu

    2016-05-01

    Diverse intercropping system has been used to control disease and improve productivity in the field. In this research, the bacterial communities in salt-alkali soils of monoculture and intercropping mulberry and soybean were studied using 454-pyrosequencing of the 16S rDNA gene. The dominant taxonomic groups were Proteobacteria, Acidobacteria, Actinobacteria, Chloroflexi, Bacteroidetes, Planctomycetes and Gemmatimonadetes and these were present across all samples. However, the diversity and composition of bacterial communities varied between monoculture and intercropping samples. The estimated bacterial diversity (H') was higher with intercropping soybean than in monoculture soybean, whereas H' showed an opposite pattern in monoculture and intercropping mulberry. Populations of Actinobacteria, Acidobacteria, and Proteobacteria were variable, depending on growth of plants as monoculture or intercropped. Most of Actinobacteria and Chloroflexi were found in intercropping samples, while Acidobacteria and Proteobacteria were present at a higher percentage in monoculture samples. The plant diversity of aboveground and microbial diversity of belowground was linked and soil pH seemed to influence the bacterial community. Finally, the specific plant species was the major factor that determined the bacterial community in the salt-alkali soils. PMID:26892826

  6. Attenuation of Endoplasmic Reticulum Stress-Mediated Liver Damage by Mulberry Leaf Diet in Streptozotocin-Induced Diabetic Rats.

    PubMed

    Afrin, Rejina; Arumugam, Somasundaram; Wahed, Mir Imam Ibne; Pitchaimani, Vigneshwaran; Karuppagounder, Vengadeshprabhu; Sreedhar, Remya; Harima, Meilei; Suzuki, Hiroshi; Miyashita, Shizuka; Nakamura, Takashi; Suzuki, Kenji; Nakamura, Masahiko; Ueno, Kazuyuki; Watanabe, Kenichi

    2016-02-01

    Endoplasmic reticulum stress (ERS) plays a crucial role in the development of insulin resistance and diabetes mellitus. Although antidiabetic use of mulberry leaves (MLs) has been popular due to their many anti-oxidative flavonoid compounds and free radical scavenging effects, ML's effects on ERS in experimental diabetic hepatocyte injury remain unknown. To investigate how ML affect ERS in diabetic liver, Sprague-Dawley (SD) rats were assigned to induce diabetes by a single intraperitoneal injection of streptozocin (STZ; 55 mg/kg) and fed with either normal chow or a diet containing 25% mulberry leaf powder diet (MLD) and examined for 56 days. We observed that MLD improved the rats' morphological and histopathological changes. Levels of ERS markers such as phosphorylated double-stranded RNA-dependent protein kinase-like endoplasmic reticulum kinase (PERK) and X-box binding protein 1 (XBP1) and the protein expression of glucose regulated protein 78 (GRP78) were significantly higher in the diabetic liver compared to normal liver. MLD for 8 weeks significantly reduced all of these markers. MLD also significantly decreased hepatocyte apoptosis, hepatic macrophage recruitment, cellular infiltration, and CCAAT/enhancer-binding protein homologous protein (CHOP), tumor necrosis factor receptor associated factor 2 (TRAF2), interleukin 1[Formula: see text] (IL-1[Formula: see text]) and sterol regulatory element binding protein isoform 1c (SREBP 1c) levels in diabetic liver. These results may suggest that MLs can preserve hepatic function in experimental diabetes by modulating ERS mediated apoptosis and liver damage. PMID:26916916

  7. Promotion of mitotic catastrophe via activation of PTEN by paclitaxel with supplement of mulberry water extract in bladder cancer cells.

    PubMed

    Chen, Nien-Cheng; Chyau, Charng-Cherng; Lee, Yi-Ju; Tseng, Hsien-Chun; Chou, Fen-Pi

    2016-01-01

    Paclitaxel is a mitotic inhibitor used in cancer chemotherapy. Mulberry fruit is rich in phenolic compounds and flavonoids and exhibits chemopreventive activities. In this study, mulberry water extract (MWE) was used as a supplement to synergize with the effects of paclitaxel in the treatment of the TSGH 8301 human bladder cancer cell line. Treatment with paclitaxel combined with MWE (paclitaxel/MWE) enhanced the cytotoxicity of paclitaxel and induced severe G2/M arrest, mitotic catastrophe and subsequent apoptosis, as shown by MTT assay, HE staining and flow cytometry analyses. Differences in the expression and activation of Aurora A and Plk1between cells treated with paclitaxel/MWE and paclitaxel alone suggested that the combined treatment caused a defect in the early steps of cytokinesis. Paclitaxel/MWE decreased EEA1immunofluorescence staining and increased the expression of PTEN, indicating that the regimen inhibited the formation of the recycling endosome, which is required for cytokinesis. Paclitaxel/MWE also retarded tumor growth in a TSGH 8301 xenograft model via activation of PTEN and Caspase 3. These data demonstrated a synergistic effect on the anticancer efficacy of paclitaxel through MWE supplementation by promoting mitotic catastrophe through the activation of PTEN, providing a novel and effective therapeutic option for bladder cancer treatment strategies. PMID:26838546

  8. Separation and identification of anthocyanin extracted from mulberry fruit and the pigment binding properties toward human serum albumin.

    PubMed

    Sheng, Feng; Wang, Yuning; Zhao, Xingchen; Tian, Na; Hu, Huali; Li, Pengxia

    2014-07-16

    Purple pigments were isolated from mulberry extracts using preparative high-speed countercurrent chromatography (HSCCC) and identified by ESI-MS/MS and high performance liquid chromatography (HPLC) techniques. The solvent system containing methyl tert-butyl ether, 1-butanol, acetonitrile, water, and trifluoroacetic acid (10:30:10:50:0.05; %, v/v) was developed in order to separate anthocyanins with different polarities. Cyanidin 3-O-(6″-O-α-rhamnopyranosyl-β-galactopyranoside) (also known as keracyanin) is the major component present in mulberry (41.3%). Other isolated pigments are cyanidin 3-O-(6″-O-α-rhamnopyranosyl-β-glucopyranoside) and petunidin 3-O-β-glucopyranoside. The binding characteristics of keracyanin with human serum albumin (HSA) were investigated by fluorescence and circular dichroism (CD) spectroscopy. Spectroscopic analysis reveals that HSA fluorescence quenched by keracyanin follows a static mode. Binding of keracyanin to HSA mainly depends on van der Waals force or H-bonds with average binding distance of 2.82 nm. The results from synchronous fluorescence, three-dimensional fluorescence, and CD spectra show that adaptive structure rearrangement and decrease of α-helical structure occur in the presence of keracyanin. PMID:24930424

  9. Differential responses in photosynthesis, growth and biomass yields in two mulberry genotypes grown under elevated CO2 atmosphere.

    PubMed

    Sekhar, Kalva Madhana; Sreeharsha, Rachapudi Venkata; Reddy, Attipalli Ramachandra

    2015-10-01

    This study was aimed to examine the responses of two mulberry genotypes (Morus alba L.), which include a drought tolerant (DT) Selection-13 (S13) and a drought susceptible (DS) Kanva-2 (K2) grown under elevated atmospheric CO2 concentration ([CO2]) of 550 μmol mol(-1). Although both genotypes exhibited positive responses to elevated CO2, S13 showed higher light saturated photosynthetic rates (A') and apparent quantum efficiency (AQE), suggesting better Rubisco carboxylation. Increased water use efficiency (WUEi) in elevated CO2 grown S13 (ES13) was due to reduced stomatal conductance (gs) and transpiration (E). Elevated CO2 significantly increased chlorophyll a fluorescence characteristics including maximum quantum yield of primary photochemistry (FV/FM) and performance index (PI(ABS)) suggesting an improved photosystem-II efficiency in both genotypes compared to their respective controls. Even though ES13 showed superior photosynthetic performance, accumulation of soluble and insoluble sugars (starch) were significantly low compared to elevated CO2 grown K2 (EK2), demonstrating higher sink capacity in ES13, which in turn resulted in better biomass yields. We conclude that S13 could be a potential genotype for mulberry-based short rotation forestry (SRF) to mitigate increasing atmospheric [CO2] as well as for the production of carbon neutral renewable bio-energy. PMID:26298067

  10. Genome-Wide Identification and Characterization of Long Non-Coding RNAs from Mulberry (Morus notabilis) RNA-seq Data

    PubMed Central

    Song, Xiaobo; Sun, Liang; Luo, Haitao; Ma, Qingguo; Zhao, Yi; Pei, Dong

    2016-01-01

    Numerous sources of evidence suggest that most of the eukaryotic genome is transcribed into protein-coding mRNAs and also into a large number of non-coding RNAs (ncRNAs). Long ncRNAs (lncRNAs), a group consisting of ncRNAs longer than 200 nucleotides, have been found to play critical roles in transcriptional, post-transcriptional, and epigenetic gene regulation across all kingdoms of life. However, lncRNAs and their regulatory roles remain poorly characterized in plants, especially in woody plants. In this paper, we used a computational approach to identify novel lncRNAs from a published RNA-seq data set and analyzed their sequences and expression patterns. In total, 1133 novel lncRNAs were identified in mulberry, and 106 of these lncRNAs displayed a predominant tissue-specific expression in the five major tissues investigated. Additionally, functional predictions revealed that tissue-specific lncRNAs adjacent to protein-coding genes might play important regulatory roles in the development of floral organ and root in mulberry. The pipeline used in this study would be useful for the identification of lncRNAs obtained from other deep sequencing data. Furthermore, the predicted lncRNAs would be beneficial towards an understanding of the variations in gene expression in plants. PMID:26938562

  11. Effects of types and amounts of stabilizers on physical and sensory characteristics of cloudy ready-to-drink mulberry fruit juice

    PubMed Central

    Akkarachaneeyakorn, Suthida; Tinrat, Sirikhwan

    2015-01-01

    In this study, the pH of mulberry juice was optimized for high anthocyanin content and an attractive red color. Mulberry juice pH values of 2.5, 4.0, 6.0, and 8.0 were evaluated. A pH of 2.5 gave an anthocyanin content of 541.39 ± 106.43 mg of cyanidin-3-glucoside per liter, and the a* value was 14 ± 1.00. The effects of stabilizers (CMC and xanthan gum) on the physical characteristics of cloudy ready-to-drink mulberry fruit juice (via the addition of mulberry fruit pulp at a mass fraction of 5%) during storage (4°C for 1 week) were also determined using different mass fractions of the stabilizers (0.1%, 0.3%, and 0.5%). Increasing the stabilizer mass fraction increased the viscosity, turbidity, stability of turbidity, and h* value. Using xanthan gum as the stabilizer produced better results for these parameters than CMC. The type of stabilizer and its mass fraction had no effect on most sensory characteristics, including appearance, color, taste, texture, and overall acceptability (P ≥ 0.05), but did affect the odor (P ≥ 0.05). Xanthan gum stabilizer gave the juice a better odor than CMC. Cloudy mulberry juice containing 0.5% xanthan gum as the stabilizer had the highest acceptance rate among panelists (average acceptance was 6.90 ± 1.37 points) and produced no precipitate during storage. PMID:25987996

  12. 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.

  13. Identification of Core Alpha 1,3-Fucosyltransferase Gene From Silkworm: An Insect Popularly Used to Express Mammalian Proteins

    PubMed Central

    Minagawa, Sachi; Sekiguchi, Satoshi; Nakaso, Yuzuru; Tomita, Masahiro; Takahisa, Manabu; Yasuda, Hideyo

    2015-01-01

    Silkworm has great potential as production system of recombinant mammalian proteins. When the protein products are used for medical purpose, it is required to reduce the risk of an allergy, the content of core alpha 1,3-fucosyl residue attached to the N-glycan of proteins, for example. We isolated the gene of an enzyme responsible for the transfer of core alpha 1,3-fucosyl residue, core alpha 1,3-fucosyltransferase (Fuc-T C3), from silkworm. A candidate cDNA for silkworm Fuc-T C3 was isolated as a homolog of the fruit fly enzyme gene fucTA. The gene was located on chromosome 7 of the silkworm genome and was composed of seven exons, which spanned approximately 10 kb on the genome. The coding region of the gene was 1,350 bp and encoded a 450-amino acid protein with a molecular mass of 52.2 kDa. Deduced amino acid sequence of the coding region showed one transmembrane domain in its N-terminal and typical motifs common to fucosyltransferases including Fuc-T C3s of other organisms in its C-terminal. The extract of CHO cells transfected with the cDNA showed Fuc-T C3 activity using GDP-fucose and DABS-GnGn peptide as substrates. These results showed this cDNA clone actually encodes silkworm Fuc-T C3. PMID:26223947

  14. 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

  15. In Vitro Evaluation of a Novel Non-Mulberry Silk Scaffold for Use in Tendon Regeneration

    PubMed Central

    Naot, Dorit; Chhana, Ashika; Matthews, Brya G.; McIntosh, Julie D.; Lin, Sandy T.C.; Choi, Ally J.; Callon, Karen E.; Dunbar, P. Rod; Lesage, Stephanie; Coleman, Brendan; Cornish, Jillian

    2015-01-01

    Tearing of the rotator cuff tendon in the shoulder is a significant clinical problem, with large/full-thickness tears present in ∼22% of the general population and recurrent tear rates postarthroscopic repair being quoted as high as 94%. Tissue-engineered biomaterials are increasingly being investigated as a means to augment rotator cuff repairs, with the aim of inducing host cell responses to increase tendon tissue regeneration. Silk-derived materials are of particular interest due to the high availability, mechanical strength, and biocompatibility of silks. In this study, Spidrex®, a novel knitted, non-mulberry silk fibroin scaffold was evaluated in vitro for its potential to improve tendon regeneration. Spidrex was compared with a knitted Bombyx mori silk scaffold, a 3D collagen gel and Fiberwire® suture material. Primary human and rat tenocytes successfully adhered to Spidrex and significantly increased in number over a 14 day period (p<0.05), as demonstrated by fluorescent calcein-AM staining and alamarBlue® assays. A similar growth pattern was observed with human tenocytes cultured on the B. mori scaffold. Morphologically, human tenocytes elongated along the silk fibers of Spidrex, assuming a tenocytic cell shape, and were less circular with a higher aspect ratio compared with human tenocytes cultured on the B. mori silk scaffold and within the collagen gel (p<0.05). Gene expression analysis by real-time PCR showed that rat tenocytes cultured on Spidrex had increased expression of tenocyte-related genes such as fibromodullin, scleraxis, and tenomodulin (p<0.05). Expression of genes that indicate transdifferentiation toward a chondrocytic or osteoblastic lineage were significantly lower in tenocytes cultured on Spidrex in comparison to the collagen gel (p<0.05). Immunogenicity assessment by the maturation of and cytokine release from primary human dendritic cells demonstrated that Spidrex enhanced dendritic cell maturation in a similar manner to the clinically used suture material Fiberwire, and significantly upregulated the release of proinflammatory cytokines (p<0.05). This suggests that Spidrex may induce an early immune response postimplantation. While further work is required to determine what effect this immune response has on the tendon healing process, our in vitro data suggests that Spidrex may have the cytocompatibility and bioactivity required to support tendon regeneration in vivo. PMID:25604072

  16. Identification and Partial Characterization of Midgut Proteases in the Lesser Mulberry Pyralid, Glyphodes pyloalis

    PubMed Central

    Mahdavi, Atiyeh; Ghadamyari, Mohammad; Sajedi, Reza H.; Sharifi, Mahbobeh; Kouchaki, Behrooz

    2013-01-01

    Proteolytic activities in digestive system extracts from the larval midgut of the lesser mulberry pyralid, Glyphodes pyloalis Walker (Lepidoptera: Pyralidae), were analyzed using different specific peptide substrates and proteinase inhibitors. High proteolytic activities were found at pH 10.0 and a temperature of 50° C using azocasein as substrate. The trypsin was active in the pH range of 9.5– 12.0, with its maximum activity at pH 11.5. Ethylene diamine tetraacetic acid had the most inhibitory effect, and 44% inhibition was detected in the presence of this inhibitor. Phenyl methane sulfonyl floride and N-tosyl-L-phe chloromethyl ketone also showed considerable inhibition of larval azocaseinolytic activity, with 40.2 and 35.1% inhibition respectively. These data suggest that the midgut of larvae contains mainly metalloproteases and serine proteases, mainly chymotrypsin. The effect of several metal ions on the activity of proteases showed that NaCl, CaCl2, CoCl2 (5 and 10 mM), and MnCl2 (5mM) reduced the protease activity. The kinetic parameters of trypsin-like proteases using N-benzoyl-L-arg-p-nitroanilide as substrate indicated that the Km and Vmax values of trypsin in the alimentary canal were 50.5 ± 2.0 µM and 116.06 ± 1.96 nmol min-1 mg-1 protein, respectively. Inhibition assays showed only small amounts of cysteine proteases were present in the G. pyloalis digestive system. The midgut digestive protease system of G. pyloalis is as diverse as that of any of the other polyphagous lepidopteran insect species, and the midgut of larvae contains mainly metalloproteases. Moreover, serine proteases and chymotrypsin also play main roles in protein digestion. Characterization of the proteolytic properties of the digestive enzymes of G. pyloalis offers an opportunity for developing appropriate and effective pest management strategies via metalloproteases and chymotrypsin inhibitors. PMID:24228902

  17. Unavailability of wild relatives

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The unavailability of crop wild relatives may come in many forms, including limited possibilities of gene flow with related species due to clonality, differing ploidy levels, or other crossing barriers between species. Alternatively, it may simply mean that we lack information about the wild relativ...

  18. Project Wild (Project Tame).

    ERIC Educational Resources Information Center

    Siegenthaler, David

    For 37 states in the United States, Project Wild has become an officially sanctioned, distributed and funded "environemtnal and conservation education program." For those who are striving to implement focused, sequential, learning programs, as well as those who wish to promote harmony through a non-anthropocentric world view, Project Wild may…

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

    PubMed Central

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

    2014-01-01

    Human growth hormone (hGH) is a peptide hormone secreted by eosinophils of the human anterior pituitary, and a regulatory factor for a variety of metabolic pathways. A 30-kD protein from the pupa stage of silkworm was detected by Western blotting and confirmed by immunoprecipitation based on its ability to bind to anti-hGH antibody. This protein, named BmhGH-like protein, was purified from fresh silkworm pupas through low-temperature homogenization, filtration, and centrifugation to remove large impurity particles. The supernatants were precipitated, resuspended, and passed through a molecular sieve. Further purification by affinity chromatography and two-dimensional electrophoresis resulted in pure protein for analysis by MS MALDI-TOF-MS analysis. An alignment with predicted proteins indicated that BmhGH-like protein consisted of two lipoproteins, which we named hGH-L1 and hGH-L2. These proteins belong to the β-trefoil superfamily, with β domains similar to the spatial structure of hGH. Assays with K562 cells demonstrated that these proteins could promote cell division in vitro. To further validate the growth-promoting effects, hGH-L2 was cloned from pupa cDNA to create recombinant silkworm baculovirus vBmNPV-hGH-L2, which was used to infect silkworm BmN cells at low titer. Flow cytometric analysis demonstrated that the protein shortened the G0/G1 phase of the cells, and enabled the cells to rapidly traverse the G1/S phase transition point to enter S phase and promote cell division. Discovery of hGH-like protein in silkworm will once again arouse people’s interest in the potential medicinal value of silkworm and establish the basis for the development of new hormone drugs. PMID:25469649

  20. Soil stability characteristics of mulberry lands at hydro-fluctuation belt in the Three Gorges Reservoir area, China.

    PubMed

    Jiang, Ping; Shi, Dongmei; Hu, Xueqin; Huang, Xianzhi; Li, Yexin; Guo, Tianlei

    2015-10-01

    The hydro-fluctuation belt in the Three Gorges Reservoir area is a typical seasonal and artificial wetland system and ecologically fragile zone. Using the widely existing mulberry forest lands in the hydro-fluctuation belt as an example and the 180-m water-level forest land as a control, this paper analyzes the soil stability of mulberry forestlands at different water levels in the hydro-fluctuation belt by analyzing and comparing the changes between soil physical and mechanical properties. The results indicated that (1) water-level changes, such as rising, flooding, draining, and exposure, affect the soil structure in mulberry forestlands. The soil agglomeration statuses for the soil layers decreased from 180 > 175 > 170 > 165 m, and the soil agglomeration statuses at a depth of 0∼20 cm decreased by 43.79, 44.95, and 57.45% compared with the control. (2) The soil water stability index decreased as follows: 180 > 170 > 175 > 165 m, which only accounted for 50.00, 47.73, and 40.91% of the control. In addition, the soil water stability indexes for the topsoils at various water levels were 1.87 (180 m), 1.67 (175 m), 2.92 (170 m), and 1.86 (165 m) times greater than those of the subsoils; thus, the resistance to hydraulic dispersion and disintegration were greater in the topsoil than in the subsoil. (3) The soil aggregate stability index decreased from 180 > 165 > 170 > 175 m and by 22.75, 23.53, and 35.29% compared with the control. (4) The soil shear strengths (composed of the cohesive force C and the internal friction angle φ) of the topsoils at water levels of 175, 170, and 165 m were significantly lower than in the control, and the internal friction angles decreased by 10.52, 19.08, and 43.25% and the cohesive force decreased by 9.88, 16.36, and 27.51%, respectively. The stability of the soil structure was greatly influenced by the soil clay content, soil organic matter content, and waterlogging duration. The study results could provide scientific support for soil and water conservation in the hydro-fluctuation belt and for biological filter construction in the Three Gorges Reservoir area to control the transport of sediment and non-point source pollutants. PMID:26385473

  1. Evaluation of bioactive compounds of black mulberry juice after thermal, microwave, ultrasonic processing, and storage at different temperatures.

    PubMed

    Jiang, Bo; Mantri, Nitin; Hu, Ya; Lu, Jiayin; Jiang, Wu; Lu, Hongfei

    2015-07-01

    The effect of different sterilization methods (thermal, microwave, and ultrasonic processing) on the main bioactive compounds and antioxidant activity of black mulberry juice during selected storage time (8 days) and temperatures (5, 15, and 25 ℃) was investigated. The antioxidant activity of thermal-treated juice depleted with storage time, whilst both ultrasound- and microwave-treated juices showed transient increase in antioxidant activity during the first 2 days that later decreased with storage time. Lower temperature storage preserved more bioactive compounds and antioxidant activity, especially in ultrasound sterilized samples. The activation energy values were 15.99, 13.07, and 12.81 kJ/mol for ultrasonic, microwave, and thermal pasteurization processes, respectively. In general, ultrasound-sterilized samples showed higher total phenolics, anthocyanin, and antioxidant activity compared to the microwave- and thermal-processed juice during the storage time especially at lower temperatures. PMID:24917651

  2. Identification and analysis of Toll-related genes in the domesticated silkworm, Bombyx mori.

    PubMed

    Cheng, Ting-Cai; Zhang, Yu-Li; Liu, Chun; Xu, Ping-Zhen; Gao, Zhi-Hong; Xia, Qing-You; Xiang, Zhong-Huai

    2008-01-01

    Silkworm (Bombyx mori), a model system for Lepidoptera, has contributed enormously to the study of insect immunology especially in humoral immunity. But little is known about the molecular mechanism of immune response in the silkworm. Toll receptors are a group of evolutionarily ancient proteins, which play a crucial role in the innate immunity of both insects and vertebrates. In human, Toll-like receptors (TLRs) are the typical pattern recognition receptors for different kinds of pathogen molecules. Toll-related receptors in Drosophila, however, were thought to function as cytokine receptors in immune response and embryogenesis. We have identified 11 putative Toll-related receptors and two Toll analogs in the silkworm genome. Phylogenetic analysis of insect Toll family and human TLRs showed that BmTolls is grouped with Drosophila Tolls and Anopheles Tolls. These putative proteins are typical transmembrane receptors flanked by the extracellular leucine-rich repeat (LRR) domain and the cytoplasmic TIR domain. Structural prediction of the TIR domain alignment found five stranded sheets and five helices, which are alternatingly joined. Microarray data indicated that BmToll and BmToll-2 were expressed with remarkable enrichment in the ovary, suggesting that they might play a role in the embryogenesis. However, the enriched expression of BmToll-2 and -4 in the midgut suggested that the proteins they encode may be involved in immune defense. Testis-specific expression of BmToll-10 and -11 and BmToLK-2 implies that these may be involved in sex-specific biological functions. The RT-PCR results indicated that 10 genes were induced or suppressed with different degrees after their immune system was challenged by different invaders. Expression profiles of BmTolls and BmToLKs reported here provide insight into their role in innate immunity and development. PMID:17499357

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

    PubMed Central

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

    2016-01-01

    The gut microbiota has a crucial role in the growth, development and environmental adaptation in the host insect. The objective of our work was to investigate the microbiota of the healthy silkworm Bombyx mori gut and changes after the infection of B. mori cypovirus (BmCPV). Intestinal contents of the infected and healthy larvae of B. mori of fifth instar were collected at 24, 72 and 144 h post infection with BmCPV. The gut bacteria were analyzed by pyrosequencing of the 16S rRNA gene. 147(135) and 113(103) genera were found in the gut content of the healthy control female (male) larvae and BmCPV-infected female (male) larvae, respectively. In general, the microbial communities in the gut content of healthy larvae were dominated by Enterococcus, Delftia, Pelomonas, Ralstonia and Staphylococcus, however the abundance change of each genus was depended on the developmental stage and gender. Microbial diversity reached minimum at 144 h of fifth instar larvae. The abundance of Enterococcus in the females was substantially lower and the abundance of Delftia, Aurantimonas and Staphylococcus was substantially higher compared to the males. Bacterial diversity in the intestinal contents decreased after post infection with BmCPV, whereas the abundance of both Enterococcus and Staphylococcus which belongs to Gram-positive were increased. Therefore, our findings suggested that observed changes in relative abundance was related to the immune response of silkworm to BmCPV infection. Relevance analysis of plenty of the predominant genera showed the abundance of the Enterococcus genus was in negative correlation with the abundance of the most predominant genera. These results provided insight into the relationship between the gut microbiota and development of the BmCPV-infected silkworm. PMID:26745627

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

    PubMed

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

    2016-01-01

    The gut microbiota has a crucial role in the growth, development and environmental adaptation in the host insect. The objective of our work was to investigate the microbiota of the healthy silkworm Bombyx mori gut and changes after the infection of B. mori cypovirus (BmCPV). Intestinal contents of the infected and healthy larvae of B. mori of fifth instar were collected at 24, 72 and 144 h post infection with BmCPV. The gut bacteria were analyzed by pyrosequencing of the 16S rRNA gene. 147(135) and 113(103) genera were found in the gut content of the healthy control female (male) larvae and BmCPV-infected female (male) larvae, respectively. In general, the microbial communities in the gut content of healthy larvae were dominated by Enterococcus, Delftia, Pelomonas, Ralstonia and Staphylococcus, however the abundance change of each genus was depended on the developmental stage and gender. Microbial diversity reached minimum at 144 h of fifth instar larvae. The abundance of Enterococcus in the females was substantially lower and the abundance of Delftia, Aurantimonas and Staphylococcus was substantially higher compared to the males. Bacterial diversity in the intestinal contents decreased after post infection with BmCPV, whereas the abundance of both Enterococcus and Staphylococcus which belongs to Gram-positive were increased. Therefore, our findings suggested that observed changes in relative abundance was related to the immune response of silkworm to BmCPV infection. Relevance analysis of plenty of the predominant genera showed the abundance of the Enterococcus genus was in negative correlation with the abundance of the most predominant genera. These results provided insight into the relationship between the gut microbiota and development of the BmCPV-infected silkworm. PMID:26745627

  5. Mulberry leaf extract mediated synthesis of gold nanoparticles and its anti-bacterial activity against human pathogens

    NASA Astrophysics Data System (ADS)

    Adavallan, K.; Krishnakumar, N.

    2014-06-01

    Gold nanoparticles (Au-NPs) were synthesized at room temperature using Morus alba (mulberry) leaf extract as reducing and stabilizing agent. The development of plant mediated synthesis of nanoparticles is gaining importance due to its simplicity, low cost, non-toxicity, eco-friendliness, long term stability and reproducible aqueous synthesis method to obtain a self-assembly of nearly monodispersed Au-NPs. The formation and morphology of biosynthesized nanoparticles are investigated with the help of UV-Vis spectroscopy, dynamic light scattering (DLS), transmission electron microscopy (TEM), atomic force microscopy (AFM), x-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FT-IR) techniques. Au-NPs formation was screened by UV-Vis spectroscopy through color conversion due to surface plasmon resonance band at 538 nm for Au-NPs. DLS studies revealed that the average size of Au-NPs was 50 nm. TEM studies showed the particles to be nearly spherical with few irregular shapes and particle size ranges 15-53 nm. The AFM image clearly shows the surface morphology of the well-dispersed Au-NPs with less than 50 nm. The high crystallinity of nanoparticles is evident from bright circular spots in the selected area electron diffraction (SAED) pattern. X-ray diffraction pattern showed high purity and face-centered cubic structure of Au-NPs. The FT-IR results indicate the presence of different functional groups present in the biomolecule capping the nanoparticles. Further, biosynthesized Au-NPs show strong zone of inhibition against Vibrio cholera (gram-negative) and Staphylococcus aureus (gram-positive) whereas, chemically synthesized Au-NPs and mulberry leaf extract exhibit a fair zone of inhibition.

  6. Mulberry Extracts Alleviate Aβ25–35-Induced Injury and Change the Gene Expression Profile in PC12 Cells

    PubMed Central

    Song, Nan; Yang, Hongpeng; Pang, Wei; Qie, Zhiwei; Lu, Hao; Tan, Long; Li, Haiqiang; Sun, Shoudan; Lian, Fuzhi; Qin, Chuan; Jiang, Yugang

    2014-01-01

    Mulberry, which contained high amounts of anthocyanins, has been used in traditional Chinese medicine. Mulberry fruit extracts (ME) have demonstrated the antioxidant activity and neuroprotection. The study was to investigate the neuroprotective efficacy of ME against β-amyloid 25–35- (Aβ25–35-) induced PC12 cells injury. Cells preincubated with or without ME (200 μg/mL) for 24 h were treated with Aβ25–35 (20 μmol/L) for another 24 h. Cell viability was assessed by MTT, gene expression profiles were examined by cDNA microarrays, and RT-PCR were used to confirm the results of microarray assays. ME pretreatment was found to neutralize the cytotoxicity and prevent Aβ25–35-induced cells injury. Analyses of gene expression profile revealed that genes involving cell adhesion, peptidase activity, cytokine activity, ion binding activity, and angiogenesis regulation were significantly modulated by ME pretreatment. Among those genes, Apaf1, Bace2, and Plcb4 were enriched in the “Alzheimer's disease-reference pathway” and downregulated after ME intervention. RT-PCR results showed that ME preincubation could significantly inhibit Aβ25–35 increased mRNA levels of these three genes. Overall, ME pretreatment could substantially alleviate PC12 cells injury and downregulate expression of AD-related genes, such as Apaf1, Bace2, and Plcb4. This study has a great nutrigenomics interest and brings new and important light in the field of AD intervention. PMID:25580148

  7. Effect of Maturity on Phenolics (Phenolic Acids and Flavonoids) Profile of Strawberry Cultivars and Mulberry Species from Pakistan

    PubMed Central

    Mahmood, Tahir; Anwar, Farooq; Abbas, Mateen; Saari, Nazamid

    2012-01-01

    In this study, we investigated how the extent of ripeness affects the yield of extract, total phenolics, total flavonoids, individual flavonols and phenolic acids in strawberry and mulberry cultivars from Pakistan. In strawberry, the yield of extract (%), total phenolics (TPC) and total flavonoids (TFC) ranged from 8.5–53.3%, 491–1884 mg gallic acid equivalents (GAE)/100 g DW and 83–327 mg catechin equivalents (CE)/100 g DW, respectively. For the different species of mulberry the yield of extract (%), total phenolics and total flavonoids of 6.9–54.0%, 201–2287 mg GAE/100 g DW and 110–1021 mg CE/100 g DW, respectively, varied significantly as fruit maturity progressed. The amounts of individual flavonols and phenolic acid in selected berry fruits were analyzed by RP-HPLC. Among the flavonols, the content of myricetin was found to be high in Morus alba (88 mg/100 g DW), the amount of quercetin as high in Morus laevigata (145 mg/100 g DW) while kaempferol was highest in the Korona strawberry (98 mg/100 g DW) at fully ripened stage. Of the six phenolic acids detected, p-hydroxybenzoic and p-coumaric acid were the major compounds in the strawberry. M. laevigata and M. nigra contained p-coumaric acid and vanillic acid while M. macroura and M. alba contained p-hydroxy-benzoic acid and chlorogenic acid as the major phenolic acids. Overall, a trend to an increase in the percentage of extraction yield, TPC, TFC, flavonols and phenolic acids was observed as maturity progressed from un-ripened to fully-ripened stages. PMID:22605997

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

    PubMed

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

    2016-01-01

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

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

    PubMed Central

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

    2016-01-01

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

  10. Wild Ponies on Assateague Island

    Wild ponies on Assateague Island. Wild ponies have lived on Assateague since the 1600s, although how they were introduced to Assateague is still debated. There are now around 300 or so wild ponies in Maryland and Virginia....

  11. A Wild Pony of Assateague

    A wild pony on Assateague Island. Wild ponies have lived on Assateague since the 1600s, although how they were introduced to Assateague is still debated. There are now around 300 or so wild ponies in Maryland and Virginia....

  12. The Wild Bunch.

    ERIC Educational Resources Information Center

    Booth, Bibi; Brook, Richard; Tisdale, Mary; Wooster, Elizabeth

    2001-01-01

    Summarizes the history of wild horses in North America and explains the social structure of horses. Discusses issues related to wildlife management. Presents activities for classroom use and includes a list of references and resources. (YDS)

  13. Mulberry leaf extract inhibits the development of atherosclerosis in cholesterol-fed rabbits and in cultured aortic vascular smooth muscle cells.

    PubMed

    Chan, Kuei-Chuan; Yang, Mon-Yuan; Lin, Ming-Cheng; Lee, Yi-Ju; Chang, Wen-Chun; Wang, Chau-Jong

    2013-03-20

    This study used high-cholesterol-fed New Zealand white rabbits and aortic vascular smooth muscle cells (VSMCs) to investigate the impact of mulberry leaf extract (MLE) on the development of atherosclerosis. The results show that the major components of MLE are polyphenols, flavonoids, carbohydrates, proteins, and lipids, and the major contituents of mulberry leaf polyphenol extract (MLPE) are polyphenols and flavonoids. In addition to improvement of liver function, the atheroma burden and levels of serum cholesterol, triglycerides, and low-density lipoprotein (LDL) are also significantly reduced after MLE treatment. MLE and MLPE improved endothelial function, inhibited proliferation and migration of aortic VSMCs, and reduced atheromas in the vascular wall. In conclusion, this study demonstrates that, in addition to exerting hypolipidemic effects, MLE and MLPE can effectively inhibit proliferation and migration of aortic VSMCs, improve vascular endothelial function, and reduce atheroma burden, thereby preventing atherosclerosis. PMID:23428158

  14. Effects of dietary mulberry, Korean red ginseng, and banaba on glucose homeostasis in relation to PPAR-alpha, PPAR-gamma, and LPL mRNA expressions.

    PubMed

    Park, Mi-Young; Lee, Kwang-Seung; Sung, Mi-Kyung

    2005-11-12

    Despite lack of scientific evidences to support its therapeutic efficacy, the use of herbal supplements has significantly increased. The purpose of this study was to evaluate the effects of traditional anti-diabetic herbs on the progress of diabetes in db/db mice, a typical non-insulin-dependent model. Five different experimental diets were as follows: control diet, 0.5% mulberry leaf water extract diet, 0.5% Korean red ginseng diet, 0.5% banaba leaf water extract diet, and 0.5% combination diet (mulberry leaf water extract/Korean red ginseng/banaba leaf water extract, 1:1:1). Blood levels of glucose, insulin, HbA1c, and triglyceride were measured every 2 weeks. At 12 weeks of age, animals were sacrificed, and tissue mRNA levels of PPAR-alpha, PPAR-gamma, and LPL were determined. Results indicated that mulberry leaf water extract, Korean red ginseng, banaba leaf water extract, and the combination of above herbs effectively reduced blood glucose, insulin, TG, and percent HbA1c in study animals (p<0.05). We also observed that the increased expressions of liver PPAR-alpha mRNA and adipose tissue PPAR-gamma mRNA in animals fed diets supplemented with test herbs. The expression of liver LPL mRNA was also increased with experimental diets containing herbs. The efficacy was highest in animals fed the combination diet for all of the markers used. These results suggest that mulberry leaf water extract, Korean red ginseng, banaba leaf water extract, and the combination of these herbs fed at the level of 0.5% of the diet significantly increase insulin sensitivity, and improve hyperglycemia possibly through regulating PPAR-mediated lipid metabolism. PMID:15979095

  15. 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

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

    PubMed

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

    2015-08-01

    The silkworm, Bombyx mori, is an economically important insect that was domesticated more than 5000 years ago. Its major economic traits focused on by breeders are quantitative traits, and an accurate and efficient QTL mapping method is necessary to explore their genetic architecture. However, current widely used QTL mapping models are not well suited for silkworm because they ignore female achiasmate and gender effects. In this study, we propose a composite method combining rational population selection and special mapping methods to map QTL in silkworm. By determining QTL for cocoon shell weight (CSW), we demonstrated the effectiveness of this method. In the CSW mapping process, only 56 markers were used and five loci or chromosomes were detected, more than in previous studies. Additionally, loci on chromosomes 1 and 11 dominated and accounted for 35.10% and 15.03% of the phenotypic variance respectively. Unlike previous studies, epistasis was detected between loci on chromosomes 11 and 22. These mapping results demonstrate the power and convenience of this method for QTL mapping in silkworm, and this method may inspire the development of similar approaches for other species with special genetic characteristics. PMID:26059330

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

    PubMed Central

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

    2015-01-01

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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  19. The large-scale production of an artificial influenza virus-like particle vaccine in silkworm pupae.

    PubMed

    Nerome, Kuniaki; Sugita, Shigeo; Kuroda, Kazumichi; Hirose, Toshiharu; Matsuda, Sayaka; Majima, Kei; Kawasaki, Kazunori; Shibata, Toshikatsu; Poetri, Okti Nadia; Soejoedono, Retno D; Mayasari, Ni L P Ika; Agungpriyono, Srihadi; Nerome, Reiko

    2015-01-01

    We successfully established a mass production system for an influenza virus-like particle (VLP) vaccine using a synthetic H5 hemagglutinin (HA) gene codon-optimized for the silkworm. A recombinant baculovirus containing the synthetic gene was inoculated into silkworm pupae. Four days after inoculation, the hemagglutination titer in homogenates from infected pupae reached a mean value of 0.8 million hemagglutination units (HAU), approximately 2,000 μg HA protein per pupa, more than 50-fold higher than that produced with an embryonated chicken egg. VLPs ranging from 30 nm to 300 nm in diameter and covered with a large number of spikes were detected in the homogenates. The spikes were approximately 14 nm long, similar to an authentic influenza HA spike. Detailed electron micrographs indicated that the VLP spike density was similar to that of authentic influenza virus particles. The results clearly show that the expression of a single HA gene can efficiently produce VLPs in silkworm pupae. When chickens were immunized with the pupae homogenate, the hemagglutination inhibition titer in their sera reached values of 2,048-8,192 after approximately 1 month. This is the first report demonstrating that a large amount of VLP vaccine could be produced by single synthetic HA gene in silkworm pupae. Our system might be useful for future vaccine development against other viral diseases. PMID:25448101

  20. 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

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

    SciTech Connect

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

    2007-04-20

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

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

    PubMed

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

    2015-01-01

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

  3. 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

  4. 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

  5. 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

  6. Identification of small auxin-up RNA (SAUR) genes in Urticales plants: mulberry (Morus notabilis), hemp (Cannabis sativa) and ramie (Boehmeria nivea).

    PubMed

    Huang, Xing; Bao, Yaning; Wang, B O; Liu, Lijun; Chen, Jie; Dai, Lunjin; Baloch, Sana Ullah; Peng, Dingxiang

    2016-03-01

    Small auxin-up RNA (SAUR) genes are important gene families in auxin signalling transduction and are commonly used as early auxin responsive markers. Till date, no SAUR gene is identified in Urticales plants despite of the published bioinformation of mulberry, hemp and ramie. In this study, we used Arabidopsis sequences as query to search against mulberry, hemp genomes and ramie transcriptome database. In total, we obtained 62, 56 and 71 SAUR genes in mulberry, hemp and ramie, respectively. Phylogenetic analysis revealed the Urticales specific expansion of SAUR genes. Expression analysis showed 15 randomly selected ramie SAUR genes that were diversely functioned in ramie tissues and revealed a series of IAA-responsive, drought-responsive and high temperature-responsive genes. Moreover, comparison of qRT-PCR data and previous RNA-Seq data suggested the reliability of our work. In this study, we first report the identification of SAUR genes in Urticales plants. These results will provide a foundation for their function validation in Urticales plant growth and development. PMID:27019439

  7. Mulberry anthocyanin extract regulates glucose metabolism by promotion of glycogen synthesis and reduction of gluconeogenesis in human HepG2 cells.

    PubMed

    Yan, Fujie; Zhang, Ji; Zhang, Lingxia; Zheng, Xiaodong

    2016-01-01

    Mulberry has been demonstrated to possess important biological activities such as antioxidation and antiinflammation. However, research on the ability of mulberry for diabetes improvement mainly focuses on the leaves and less on the fruit. This study showed that a mulberry anthocyanin extract (MAE) had a significant effect on increasing the glucose consumption in HepG2 cells. The MAE enhanced the glycogen content and suppressed levels of glucose production. The enzyme activities of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) were decreased in HepG2 cells after MAE treatment due to PPARγ coactivator 1α (PGC-1α) and forkhead box protein O1 (FOXO1) inhibition. Moreover, the phosphorylation of protein kinase B (AKT) and glycogen synthase kinase-3β (GSK-3β) was increased by the MAE, leading to an expression enhancement of glycogen synthase 2 (GYS2). And this effect was blocked by the phosphoinositide 3-kinase (PI3K) inhibitor LY294002. In summary, our results suggested that the MAE regulates glucose metabolism by activating the PI3K/AKT pathway that relates to glycogen synthesis as well as through the inhibition of key molecules that promote gluconeogenesis. PMID:26467565

  8. Detection of adulteration in mulberry pekmez samples added various sugar syrups with ¹³C/¹²C isotope ratio analysis method.

    PubMed

    Tosun, Murat

    2014-12-15

    Mulberry pekmez can be adulterated in different ways either during the production process or after production is completed. To identify these adulterations, stable carbon isotope ratio analysis (SCIRA) was performed on the model examples prepared by adding saccharose syrup (SS), glucose syrup (GS) and high fructose corn syrup (HFCS) into two different pure mulberry pekmez samples in the ratios of 0%, 10%, 30% and 50%. The δ(13)C ratio of the pure mulberry pekmez was determined as -26.60‰ on average, the saccharose syrup as -24.80‰, the glucose syrup as -11.20‰ and the high-fructose corn syrup as -11.40‰. In identifying the adulteration made to pekmez, especially with the high-fructose corn syrup, which is obtained from corn starch, and with the glucose syrup, the δ(13)C ratio comes into prominence. However it remains impossible identify the adulterations made with the saccharose, which is obtained from beet sugar, or invert sugar syrups. PMID:25038711

  9. Mulberry leaf extract inhibits vascular smooth muscle cell migration involving a block of small GTPase and Akt/NF-kappaB signals.

    PubMed

    Chan, Kuei-Chuan; Ho, Hsieh-Hsun; Huang, Chien-Ning; Lin, Ming-Cheng; Chen, Hsiang-Mei; Wang, Chau-Jong

    2009-10-14

    Mulberry, the fruit of Morus alba, is commonly used in Chinese medicines because of its many pharmacologic effects. Mulberry leaves contain many phenolic antioxidants that can reduce cardiovascular disease. Atherosclerosis involves proliferation and migration of vascular smooth muscle cell (VSMC). Thus, we investigated the mechanisms by which mulberry leaf extract (MLE) might inhibit migration of VSMC. MLE was rich in polyphenols (44.82%), including gallic acid, protocatechuic acid, catechin, gallocatechin gallate, caffeic acid, epicatechin, rutin, and quercetin. MLE could inhibit the migration of A7r5 cells in a dose- and time-dependent manner. MLE also inhibited the activities of matrix metalloproteinases (MMPs) MMP-2 and MMP-9, protein expressions, and phosphorylation of FAK and Akt, and protein expressions of small guanosine triphosphatases (GTPases: c-Raf, Ras, Rac1, Cdc42, and RhoA) in a dose-dependent manner. NF-kappaB expression was also inhibited by MLE. MLE could effectively inhibit the migration of VSMC by blocking small GTPase and Akt/NF-kappaB signals. PMID:19761240

  10. The effect of calorie restriction on growth and development in silkworm, Bombyx mori.

    PubMed

    Li, Yijia; Chen, Keping; Yao, Qin; Li, Jun; Wang, Yong; Liu, Haijun; Zhang, Chiyu; Huang, Guoping

    2009-07-01

    Caloric restriction (CR) is known to extend the life span in different species from yeast to mammals. In this report, a simple organism silkworm (Bombyx mori) was used to study the effect of moderate CR on the growth and development processes of insects. Here we show that an extension of life span upon moderate CR was observed in the silkworm. The total protein level in the 5th instar larvae hemolymph appeared to decline significantly under CR. SDS-PAGE analysis showed that the influence of CR was sex-dependent. The CR effects on female animals were much more significant than on the males. The MALDI-TOF MS study identified 16 proteins that expressed differentially among six groups of the male or female larvae fed at different time frequencies. Four of them, storage protein 1 (SP1), arylphorin (SP2), imaginal disk growth factor (IDGF), and 30-kDa lipoprotein, showed significant differences. It was demonstrated that these four proteins were up-regulated when the larvae were over-fed and down-regulated when the larvae were less-fed. PMID:19479736

  11. The silkworm W chromosome is a source of female-enriched piRNAs.

    PubMed

    Kawaoka, Shinpei; Kadota, Koji; Arai, Yuji; Suzuki, Yutaka; Fujii, Tsuguru; Abe, Hiroaki; Yasukochi, Yuji; Mita, Kazuei; Sugano, Sumio; Shimizu, Kentaro; Tomari, Yukihide; Shimada, Toru; Katsuma, Susumu

    2011-12-01

    In the silkworm, Bombyx mori, the W chromosome plays a dominant role in female determination. However, neither protein-coding genes nor transcripts have so far been isolated from the W chromosome. Instead, a large amount of functional transposable elements and their remnants are accumulated on the W chromosome. PIWI-interacting RNAs (piRNAs) are 23-30-nt-long small RNAs that potentially act as sequence-specific guides for PIWI proteins to silence transposon activity in animal gonads. In this study, by comparing ovary- and testis-derived piRNAs, we identified numerous female-enriched piRNAs. Our data indicated that female-enriched piRNAs are derived from the W chromosome. Moreover, comparative analyses on piRNA profiles from a series of W chromosome mutant strains revealed a striking enrichment of a specific set of transposon-derived piRNAs in the putative sex-determining region. Collectively, we revealed the nature of the silkworm W chromosome as a source of piRNAs. PMID:22020973

  12. Possible effect of 30K proteins in embryonic development of silkworm Bombyx mori.

    PubMed

    Zhong, Bo-Xiong; Li, Jian-Ke; Lin, Jian-Rong; Liang, Jian-She; Su, Song-Kun; Xu, Hai-Sheng; Yan, Hai-Yan; Zhang, Ping-Bo; Fujii, Hiroshi

    2005-05-01

    The silkworm Bombyx mori possesses a 30K protein family of 3x10(4) Da, the biological functions of which have not been fully identified. The relationship between the 30K protein family and the embryonic development of temperature sensitive sex-linked mutant strain of silkworm was investigated by two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The results show that protein spots 1-5 of the 30K protein family, mainly existing in normal strain, are possibly related to embryonic development. The early consumption of a 30K protein named 6G1-30K-1 and the accumulation of 30K proteins named 6G1-30K-3 and 6G1-30K-4 are likely caused by the destruction of physiological balance in normal embryonic development, which may lead to lower hatchability of the temperature sensitive strain. The results suggest that reasonable metabolism of 30K proteins is a prerequisite for the embryo's normal development. PMID:15880265

  13. Characterization of a dual-CRD galectin in the silkworm Bombyx mori.

    PubMed

    Rao, Xiang-Jun; Wu, Peng; Shahzad, Toufeeq; Liu, Su; Chen, Ling; Yang, Yun-Fan; Shi, Qiao; Yu, Xiao-Qiang

    2016-07-01

    Galectins (S-type lectins) are an ancient family of lectins with the β-galactoside binding activity. In mammals, galectins play essential roles in many biological processes, such as development, immune homeostasis and tumor progression. However, few studies have been devoted to their functions in insects. Here, we characterized the only dual-CRD galectin in the silkworm Bombyx mori (BmGalectin-4). BmGalectin-4 cDNA possesses an open reading frame of 1089 bp, which encodes a putative galectin of 363 amino acids containing tandem carbohydrate recognition domains (CRDs). BmGalectin-4 was expressed in various tissues but the protein was most abundant in fertilized eggs. Its transcript level in fertilized eggs was upregulated upon bacterial challenge. Recombinant BmGalectin-4 purified from Escherichia coli bound to bacterial cell wall components and bacterial cells. In addition, the recombinant protein induced bacterial agglutination, but did not have antibacterial activity against selected microorganisms. Taken together, our results suggest that BmGalectin-4 may function as a pattern recognition receptor primarily in silkworm fertilized eggs. PMID:26944801

  14. Existence of Prophenoloxidase in Wing Discs: A Source of Plasma Prophenoloxidase in the Silkworm, Bombyx mori

    PubMed Central

    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

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed

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

    2015-01-01

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

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

    PubMed

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

    2016-01-01

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

  18. Estimation of amino acids, urea and uric acid in tasar silkworm, Antheraea mylitta Drury.

    PubMed

    Shamitha, G; Rao, A Purushotham

    2008-11-01

    The tasar silkworm, Antheraea mylitta Drury, Andhra local ecorace is an exclusive race of Andhra Pradesh. It is on the verge of extinction due to difficulty of acclimatisation at breeding and rearing stages. As an attempt to protect this race, a method of total indoor rearing has been done. In this context, the estimation of free amino acids, excretory products- urea and uric acid were compared during the fourth and fifth instars of tasar silkworm, reared under outdoor and indoor conditions. The study has revealed that amino acids decreased in the fat body in outdoor and indoor reared larvae in contrast to that in the haemolymph where it has gradually increased from first to third crops. This is an important finding as it reveals that indoor worms seem to adopt proteolytic activity in the haemolymph. Secondly, in the fifth instar the excretory products are more compared to fourth instar in the indoor reared worms. During fifth instar, formation of nitrogenous products lessens as silk synthesis enhances. The present study reveals that decrease in uric acid in fifth instar implies increase in growth rate and silk synthesis in both outdoor and indoor worms. The findings of the present investigation is helpful in the conservation and protection of the A. mylitta, Andhra local ecorace. PMID:19297987

  19. Molecular mechanisms of novel peptides from silkworm pupae that inhibit α-glucosidase.

    PubMed

    Zhang, Yu; Wang, Nan; Wang, Wei; Wang, Junhong; Zhu, Zuoyi; Li, Xue

    2016-02-01

    The objectives of this study were to identify peptides that inhibit α-glucosidase using a quantitative structure-activity relationship (QSAR) screening method and a database of silkworm peptides. This study compared the docking characteristics of several peptides with high inhibitory activity against α-glucosidase and summarized the molecular mechanisms by which the silkworm peptides affected α-glucosidase. Four peptides that strongly inhibited α-glucosidase were obtained: Gln-Pro-Gly-Arg with IC50 at 65.8μmol/L, Ser-Gln-Ser-Pro-Ala at 20μmol/L, Gln-Pro-Pro-Thr at 560μmol/L and Asn-Ser-Pro-Arg at 205μmol/L. Studies docking the peptides to the active site of α-glucosidase (PDB ID: 2QMJ) showed that a common characteristic was Lys776 in 2QMJ, which could be a critical target for α-glucosidase trapping of inhibitory peptides. The results revealed that the four peptides, especially Ser-Gln-Ser-Pro-Ala, could be potential drugs for treating diabetes. PMID:26724364

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

    PubMed

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

    2016-01-15

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

  1. Expression of human endostatin in larvae of silkworm (Bombyx mori) and in vitro activity assays.

    PubMed

    Yongfeng, Jin; Yingfei, Wang; Zhenhong, Zhu; Yaozhou, Zhang

    2002-08-01

    Human endostatin is a novel antiangiogenic molecule, which can inhibit the proliferation and development of new blood vessels, and experimentally can cause nearly complete regression of established tumors. In this paper, the cDNA encoding human endostatin was cloned into a baculovirus shuttle vector pBacPAK8 and co-infected with linearized Bm-BacPAK6 DNA into and BmN cells. The recombinant virus was screened and identified by PCR, DNA and RNA dot hybridization, and ELISA assay. The recombinant endostatin was expressed in culture cells, and the larvae and pupa of silkworm by inoculation of recombinant virus. The biological activity assay showed that the expression product in larvae was over 150 microg/ml, about 50-fold higher than that expressed in cultured cells. SDS-PAGE and Western blotting analysis showed a pattern of molecular weight of about 20 kDa. The bio-activity of the protein product was determined by human umbilical vein endothelial cells (ECV304) proliferation test in vitro and the chick chorioallantoic membrane (CAM) vascular inhibition test. Endostatin showed significant inhibitory effect on endothelial cells in a dose-dependent manner. Silkworm-produced endostatin induced apoptosis of endothelial cells and also inhibited angiogenesis in the CAM assay. Combination regimen using angiostatin and endostatin showed more than additive effect in angiogenic inhibition and increasing apoptosis when compared with treatment with the individual antiangiogenic protein. PMID:12186748

  2. Effects of 10-hydroxycamptothecin on intrinsic mitochondrial pathway in silkworm BmN-SWU1 cells.

    PubMed

    Pan, Chun; Hu, Yan-Fen; Song, Juan; Yi, Hua-Shan; Wang, La; Yang, Yi-Ying; Wang, Yong-Ping; Zhang, Man; Pan, Min-Hui; Lu, Cheng

    2016-02-01

    10-Hydroxycamptothecin (HCPT), a plant alkaloid isolated from Camptotheca acuminate, is known as a planted-derived insecticide, however, the specific mechanism in insect cells is still unclear. In this study, we treated the ovarian cell line of the silkworm, BmN-SWU1, with different HCPT doses for durations ranging from 0 to 72h. The apoptosis morphology was evident after 72h of incubation and included cell protuberance, concentrated cytoplasm and apoptotic bodies. We observed DNA fragmentation and cell apoptosis after HCPT treatment. The disruption of mitochondrial distribution, activation of the intracellular mitochondrial permeability transition pore, and release of cytochrome c during HCPT-induced apoptosis in dose and time-dependent manner indicate the involvement of mitochondria in BmN-SWU1 cells. Caspase-9 and -3 activities increased gradually with the duration of incubation time. In conclusion, HCPT has a significant effect to initiate the intrinsic mitochondrial pathway in silkworm cells, providing a theoretical basis for better application of plant-derived insecticide in pest control. PMID:26821653

  3. 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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2015-05-22

    Insect hemocytes play essential roles in the metabolism, metamorphosis and immunity, which are closely related events of growth and development. Here, four novel hemocyte-specific genes were obtained and conformed in our study, namely, Bmintβ2, Bmintβ3, BmCatO, and BmSw04862, respectively. Subsequently, their promoter sequences were cloned, and their activity in hemocytes, fat body, and silk gland were analyzed using recombinant AcNPV vector system in vivo. Our results showed that Bmintβ2, Bmintβ3, and BmCatO were hemocyte-specific promoters in the silkworm, Bombyx mori. Interestingly, Bmintβ2, and Bmintβ3 promoter regions were both located in their first intron. Further analysis of a series of BmCatO promoter truncations showed that a 254 bp region could function as a promoter element in the tissue-specificity expression. In summary, the results of this study revealed that we have identified three hemocyte-specific promoters in silkworm that will not only great significance for better understanding of hemocyte-specific gene, but also has potential applications in insect hematopoiesis and innate immunity research. PMID:25862374

  6. Ecotoxicology of wild mammals

    USGS Publications Warehouse

    Rattner, B.A.; Shore, R.F.

    2000-01-01

    An international group of 32 scientists has critically reviewed the scientific literature on exposure and effects of environmental contaminants in wild mammals. Although the absolute number of toxicological studies in domesticated and wild mammals eclipses that for birds, a detailed examination of scientific publications and databases reveal that information for 'wild' birds is actually greater than that for 'wild' mammals. Of the various taxa of mammals, ecotoxicological data is most noticeably lacking for marsupials and monotremes. In contrast, rodents (comprising 43% of all mammal species) have been studied extensively, despite evidence of their tolerance to some organochlorine compounds, rodenticides, and even radionuclides. Mammalian species at greatest risk of exposure include those that consume a high percentage of their body weight on a daily basis (e.g., shrews, moles and bats). Aquatic mammals tend to bioaccumulate tremendous burdens of lipophilic contaminants, although storage in their fat depots may actually limit toxicity. Carnivores appear to be more sensitive to adverse effects of environmental contaminants than herbivores. Remarkably few of the thousands of compounds manufactured worldwide have been toxicologically evaluated in wild mammals, and concentrations of even fewer have been monitored in tissues. Overarching research needs include: development of new exposure/effects models and better methods for estimation of species sensitivities; generation of comparative data on contaminant bioavailability, sublethal responses and detoxication mechanisms; enhanced understanding of pesticide, industrial contaminant and metal interactions; identification of endocrine disruptive contaminants and their overall ecological significance; and finally, estimating the relative contribution of environmental contamination as a factor affecting wild mammal populations.

  7. Silkworm thermal biology: a review of heat shock response, heat shock proteins and heat acclimation in the domesticated silkworm, Bombyx mori.

    PubMed

    Manjunatha, H B; Rajesh, R K; Aparna, H S

    2010-01-01

    Heat shock proteins (HSPs) are known to play ecological and evolutionary roles in this postgenomic era. Recent research suggests that HSPs are implicated in cardiovascular biology and disease development, proliferation and regulation of cancer cells, cell death via apoptosis, and several other key cellular functions. These activities have generated great interest amongst cell and molecular biologists, and these biologists are keen to unravel other hitherto unknown potential functions of this group of proteins. Consequently, the biological significance of HSPs has led to cloning and characterization of genes encoding HSPs in many organisms including the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae). However, most of the past investigations in B. mori were confined to expression of HSPs in tissues and cell lines, whereas information on their specific functional roles in biological, physiological, and molecular processes is scarce. Naturally occurring or domesticated polyvoltines (known to be the tropical race) are more resistant to high temperatures and diseases than bi- or univoltines (temperate races). The mechanism of ecological or evolutionary modification of HSPs during the course of domestication of B. mori - particularly in relation to thermotolerance in geographically distinct races/strains - is still unclear. In addition, the heat shock response, thermal acclimation, and hardening have not been studied extensively in B. mori compared to other organisms. Towards this, recent investigations on differential expression of HSPs at various stages of development, considering the concept of the whole organism, open ample scope to evaluate their biological and commercial importance in B. mori which has not been addressed in any of the representative organisms studied so far. Comparatively, heat shock response among different silkworm races/strains of poly-, bi-, and univoltines varies significantly and thermotolerance increases as the larval development proceeds. Hence, this being the first review in this area, an attempt has been made to collate all available information on the heat shock response, HSPs expression, associated genes, amino acid sequences, and acquired/unacquired thermotolerance. The aim is to present this as a valuable resource for addressing the gap in knowledge and understanding evolutionary significance of HSPs between domesticated (B. mori) and non-domesticated insects. It is believed that the information presented here will also help researchers/breeders to design appropriate strategies for developing novel strains for the tropics. PMID:21265618

  8. Silkworm Thermal Biology: A Review of Heat Shock Response, Heat Shock Proteins and Heat Acclimation in the Domesticated Silkworm, Bombyx mori

    PubMed Central

    Manjunatha, H. B.; Rajesh, R. K.; Aparna, H. S.

    2010-01-01

    Heat shock proteins (HSPs) are known to play ecological and evolutionary roles in this postgenomic era. Recent research suggests that HSPs are implicated in cardiovascular biology and disease development, proliferation and regulation of cancer cells, cell death via apoptosis, and several other key cellular functions. These activities have generated great interest amongst cell and molecular biologists, and these biologists are keen to unravel other hitherto unknown potential functions of this group of proteins. Consequently, the biological significance of HSPs has led to cloning and characterization of genes encoding HSPs in many organisms including the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae). However, most of the past investigations in B. mori were confined to expression of HSPs in tissues and cell lines, whereas information on their specific functional roles in biological, physiological, and molecular processes is scarce. Naturally occurring or domesticated polyvoltines (known to be the tropical race) are more resistant to high temperatures and diseases than bi- or univoltines (temperate races). The mechanism of ecological or evolutionary modification of HSPs during the course of domestication of B. mori - particularly in relation to thermotolerance in geographically distinct races/strains - is still unclear. In addition, the heat shock response, thermal acclimation, and hardening have not been studied extensively in B. mori compared to other organisms. Towards this, recent investigations on differential expression of HSPs at various stages of development, considering the concept of the whole organism, open ample scope to evaluate their biological and commercial importance in B. mori which has not been addressed in any of the representative organisms studied so far. Comparatively, heat shock response among different silkworm races/strains of poly-, bi-, and univoltines varies significantly and thermotolerance increases as the larval development proceeds. Hence, this being the first review in this area, an attempt has been made to collate all available information on the heat shock response, HSPs expression, associated genes, amino acid sequences, and acquired/unacquired thermotolerance. The aim is to present this as a valuable resource for addressing the gap in knowledge and understanding evolutionary significance of HSPs between domesticated (B. mori) and non-domesticated insects. It is believed that the information presented here will also help researchers/breeders to design appropriate strategies for developing novel strains for the tropics. PMID:21265618

  9. Prevention Effects and Possible Molecular Mechanism of Mulberry Leaf Extract and its Formulation on Rats with Insulin-Insensitivity

    PubMed Central

    Xie, Chen; Luo, Xiuzhen; Bao, Yonggang; Wu, Bin; Hu, Yuchi; Zhong, Zhong; Liu, Chang; Li, MinJie

    2016-01-01

    For centuries, mulberry leaf has been used in traditional Chinese medicine for the treatment of diabetes. This study aims to test the prevention effects of a proprietary mulberry leaf extract (MLE) and a formula consisting of MLE, fenugreek seed extract, and cinnamon cassia extract (MLEF) on insulin resistance development in animals. MLE was refined to contain 5% 1-deoxynojirimycin by weight. MLEF was formulated by mixing MLE with cinnamon cassia extract and fenugreek seed extract at a 6:5:3 ratio (by weight). First, the acute toxicity effects of MLE on ICR mice were examined at 5 g/kg BW dose. Second, two groups of normal rats were administrated with water or 150 mg/kg BW MLE per day for 29 days to evaluate MLE’s effect on normal animals. Third, to examine the effects of MLE and MLEF on model animals, sixty SD rats were divided into five groups, namely, (1) normal, (2) model, (3) high-dose MLE (75 mg/kg BW) treatment; (4) low-dose MLE (15 mg/kg BW) treatment; and (5) MLEF (35 mg/kg BW) treatment. On the second week, rats in groups (2)-(5) were switched to high-energy diet for three weeks. Afterward, the rats were injected (ip) with a single dose of 105 mg/kg BW alloxan. After four more days, fasting blood glucose, post-prandial blood glucose, serum insulin, cholesterol, and triglyceride levels were measured. Last, liver lysates from animals were screened with 650 antibodies for changes in the expression or phosphorylation levels of signaling proteins. The results were further validated by Western blot analysis. We found that the maximum tolerance dose of MLE was greater than 5 g/kg in mice. The MLE at a 150 mg/kg BW dose showed no effect on fast blood glucose levels in normal rats. The MLE at a 75 mg/kg BW dose and MLEF at a 35 mg/kg BW dose, significantly (p < 0.05) reduced fast blood glucose levels in rats with impaired glucose and lipid metabolism. In total, 34 proteins with significant changes in expression and phosphorylation levels were identified. The changes of JNK, IRS1, and PDK1 were confirmed by western blot analysis. In conclusion, this study demonstrated the potential protective effects of MLE and MLEF against hyperglycemia induced by high-energy diet and toxic chemicals in rats for the first time. The most likely mechanism is the promotion of IRS1 phosphorylation, which leads to insulin sensitivity restoration. PMID:27054886

  10. Prevention Effects and Possible Molecular Mechanism of Mulberry Leaf Extract and its Formulation on Rats with Insulin-Insensitivity.

    PubMed

    Liu, Yan; Li, Xuemei; Xie, Chen; Luo, Xiuzhen; Bao, Yonggang; Wu, Bin; Hu, Yuchi; Zhong, Zhong; Liu, Chang; Li, MinJie

    2016-01-01

    For centuries, mulberry leaf has been used in traditional Chinese medicine for the treatment of diabetes. This study aims to test the prevention effects of a proprietary mulberry leaf extract (MLE) and a formula consisting of MLE, fenugreek seed extract, and cinnamon cassia extract (MLEF) on insulin resistance development in animals. MLE was refined to contain 5% 1-deoxynojirimycin by weight. MLEF was formulated by mixing MLE with cinnamon cassia extract and fenugreek seed extract at a 6:5:3 ratio (by weight). First, the acute toxicity effects of MLE on ICR mice were examined at 5 g/kg BW dose. Second, two groups of normal rats were administrated with water or 150 mg/kg BW MLE per day for 29 days to evaluate MLE's effect on normal animals. Third, to examine the effects of MLE and MLEF on model animals, sixty SD rats were divided into five groups, namely, (1) normal, (2) model, (3) high-dose MLE (75 mg/kg BW) treatment; (4) low-dose MLE (15 mg/kg BW) treatment; and (5) MLEF (35 mg/kg BW) treatment. On the second week, rats in groups (2)-(5) were switched to high-energy diet for three weeks. Afterward, the rats were injected (ip) with a single dose of 105 mg/kg BW alloxan. After four more days, fasting blood glucose, post-prandial blood glucose, serum insulin, cholesterol, and triglyceride levels were measured. Last, liver lysates from animals were screened with 650 antibodies for changes in the expression or phosphorylation levels of signaling proteins. The results were further validated by Western blot analysis. We found that the maximum tolerance dose of MLE was greater than 5 g/kg in mice. The MLE at a 150 mg/kg BW dose showed no effect on fast blood glucose levels in normal rats. The MLE at a 75 mg/kg BW dose and MLEF at a 35 mg/kg BW dose, significantly (p < 0.05) reduced fast blood glucose levels in rats with impaired glucose and lipid metabolism. In total, 34 proteins with significant changes in expression and phosphorylation levels were identified. The changes of JNK, IRS1, and PDK1 were confirmed by western blot analysis. In conclusion, this study demonstrated the potential protective effects of MLE and MLEF against hyperglycemia induced by high-energy diet and toxic chemicals in rats for the first time. The most likely mechanism is the promotion of IRS1 phosphorylation, which leads to insulin sensitivity restoration. PMID:27054886

  11. Frozen Wild River

    The USGS gages the Wild River at Gilead, Maine at station 01054200. This photo was taken during a winter trip to the site. During this trip, the gage equipment was checked and a discharge measurement was made by drilling holes through the ice and lowering velocity meters into the water....

  12. Endangered Species: Wild & Rare.

    ERIC Educational Resources Information Center

    Braus, Judy, Ed.

    1987-01-01

    Ranger Rick's NatureScope is a creative education series dedicated to inspiring in children an understanding and appreciation of the natural world while developing the skills they will need to make responsible decisions about the environment. The topic of this issue is "Endangered Species: Wild and Rare." Contents are organized into the following

  13. William Wilde: Historian.

    PubMed

    Geary, L

    2016-05-01

    This essay attempts to assess William Wilde as a social historian. It examines some of his contributions to the discipline of history and looks particularly at 'The food of the Irish', which was published in the Dublin University Magazine in February 1854. PMID:26969457

  14. Flooded Wild Rice River

    Wild Rice River at Great Bend North Dakota, streamflow 1,890 cubic feet per second.  Photograph taken during spring 2010 flooding looking downstream of the bridge which was clogged with debris.  The river also had flooded over the road approaching the bridge....

  15. Blastomycosis in wild wolves

    USGS Publications Warehouse

    Thiel, R.P.; Mech, L.D.; Ruth, G.R.; Archer, J.R.; Kaufman, L.

    1987-01-01

    Blastomycosis was fatal to a wild wolf in Minnesota, and serologic evidence of blastomycosis was found in a Wisconsin wolf. No unusual movements were detected in the Minnesota animal from October 1983 through October 1985. However, by early December 1985, this wolf was weak and debilitated, and it perished on 14 December after approaching a human residence.

  16. Taming the Wild Text

    ERIC Educational Resources Information Center

    Allyn, Pam

    2012-01-01

    As a well-known advocate for promoting wider reading and reading engagement among all children--and founder of a reading program for foster children--Pam Allyn knows that struggling readers often face any printed text with fear and confusion, like Max in the book Where the Wild Things Are. She argues that teachers need to actively create a

  17. Wild and Crafty.

    ERIC Educational Resources Information Center

    Braus, Judy, Ed.

    1988-01-01

    Ranger Rick's NatureScope is a creative education series dedicated to inspiring in children an understanding and appreciation of the natural world while developing the skills they will need to make responsible decisions about the environment. The topic of this issue is "Wild & Crafty." It contains a variety of craft ideas related to animal life…

  18. Wild and Crafty.

    ERIC Educational Resources Information Center

    Braus, Judy, Ed.

    1988-01-01

    Ranger Rick's NatureScope is a creative education series dedicated to inspiring in children an understanding and appreciation of the natural world while developing the skills they will need to make responsible decisions about the environment. The topic of this issue is "Wild & Crafty." It contains a variety of craft ideas related to animal life

  19. Marc Chagall: "Wild Poppies."

    ERIC Educational Resources Information Center

    White, Carolyn

    1987-01-01

    Based on a full-color reproduction of Marc Chagall's painting, "Wild Poppies," the goals of this lesson plan are to introduce students to artist's use of dreams and memories in making art, to communicate the idea that artists include their visual memories of people and things they love in their artwork, and to introduce the concepts of line and…

  20. Taming the Wild Text

    ERIC Educational Resources Information Center

    Allyn, Pam

    2012-01-01

    As a well-known advocate for promoting wider reading and reading engagement among all children--and founder of a reading program for foster children--Pam Allyn knows that struggling readers often face any printed text with fear and confusion, like Max in the book Where the Wild Things Are. She argues that teachers need to actively create a…

  1. Endangered Species: Wild & Rare.

    ERIC Educational Resources Information Center

    Braus, Judy, Ed.

    1987-01-01

    Ranger Rick's NatureScope is a creative education series dedicated to inspiring in children an understanding and appreciation of the natural world while developing the skills they will need to make responsible decisions about the environment. The topic of this issue is "Endangered Species: Wild and Rare." Contents are organized into the following…

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

    PubMed Central

    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 830.81050bp was significantly associated with LWT, CWT, SWT, SR and floss content, indicated its pleiotropic role. Two ISSR markers, 835.51950bp and 825.9710bp showed significant association with floss content and TLD. These markers were segregated in F2 generation and Chi-square test confirmed (χ2 = ~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

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

    PubMed Central

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

    2013-01-01

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

  4. Effects of Wolbachia in the uzifly, Exorista sorbillans, a parasitoid of the silkworm, Bombyx mori.

    PubMed

    Puttaraju, H P; Prakash, B M

    2005-01-01

    The uzifly, Exorista sorbillans (Diptera: Tachinidae), a parasitoid of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), harbours Wolbachia (Rickettsia) endosymbionts. Administration of 0.05 mg/ml oxytetracycline to the adult uziflies removed Wolbachia endosymbionts and resulted in different reproductive disorders, such as i) reduction in fecundity of uninfected females, ii) cytoplasmic incompatibility in crosses between infected males and uninfected females, iii) sterility in the crosses between both males and females from uninfected populations, and iv) sex-ratio distortion in uninfected females irrespective of the presence of Wolbachia in males. However, tetracycline treatment did not have much effect on longevity of the uzifly. These results suggest that the interaction of Wolbachia with its uzifly host is one of mutual symbiosis as it controls the reproductive physiology of its hosts. PMID:17119612

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

    PubMed

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

    2013-04-23

    Transgene-based genetic sexing methods are being developed for insects of agricultural and public health importance. Male-only rearing has long been sought in sericulture because males show superior economic characteristics, such as better fitness, lower food consumption, and higher silk yield. Here we report the establishment of a transgene-based genetic sexing system for the silkworm, Bombyx mori. We developed a construct in which a positive feedback loop regulated by sex-specific alternative splicing leads to high-level expression of the tetracycline-repressible transactivator in females only. Transgenic animals show female-specific lethality during embryonic and early larval stages, leading to male-only cocoons. This transgene-based female-specific lethal system not only has wide application in sericulture, but also has great potential in lepidopteran pest control. PMID:23569267

  6. Identification of candidate aldehyde oxidases from the silkworm Bombyx mori potentially involved in antennal pheromone degradation.

    PubMed

    Pelletier, Julien; Bozzolan, Françoise; Solvar, Marthe; François, Marie-Christine; Jacquin-Joly, Emmanuelle; Maïbèche-Coisne, Martine

    2007-12-01

    Signal inactivation is a crucial step in the dynamic of olfactory process and involves various Odorant-Degrading Enzymes. In the silkworm Bombyx mori, one of the best models for studying olfaction in insects, the involvement of an antennal-specific aldehyde oxidase in the degradation of the sex pheromone component bombykal has been demonstrated over the three past decades by biochemical studies. However, the corresponding enzyme has never been characterized at the molecular level. Bioinformatic screening of B. mori genome and molecular approaches have been used to isolate several candidate sequences of aldehyde oxidases. Two interesting antennal-expressed genes have been further characterized and their putative functions are discussed in regard to their respective expression pattern and to our knowledge on aldehyde oxidase properties. Interestingly, one gene appeared as specifically expressed in the antennae of B. mori and associated in males with the bombykal-sensitive sensilla, strongly suggesting that it could encode for the previously biochemically characterized enzyme. PMID:17904312

  7. Transcription factor SGF1 is critical for the neurodevelopment in the silkworm, Bombyx mori.

    PubMed

    Liu, Zhao-Yang; Yu, Qi; Yang, Chun-Hong; Meng, Miao; Ren, Chun-Jiu; Mu, Zhi-Mei; Cui, Wei-Zheng; Liu, Qing-Xin

    2016-08-01

    FoxA transcription factors play vital roles in regulating the expression of organ-specific genes. BmSGF1, the sole FoxA family member in Bombyx mori, is required for development of the silk gland. However, the function of BmSGF1 in development of the nervous system in the silkworm remains unknown. Here, we show that the amino acids sequence of BmSGF1 is evolutionarily conserved in its middle region from Trichoplax adhaerens to human and diverged from the homologues in most other species in its N-terminal region. BmSGF1 expresses in the nervous system at the embryonic stage. Knockdown of Bmsgf1 by RNA interference (RNAi) results in abnormal development of axons. Therefore, our results demonstrate that BmSGF1 is an indispensable regulator for neurodevelopment. PMID:27106119

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

    PubMed Central

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

    2013-01-01

    Transgene-based genetic sexing methods are being developed for insects of agricultural and public health importance. Male-only rearing has long been sought in sericulture because males show superior economic characteristics, such as better fitness, lower food consumption, and higher silk yield. Here we report the establishment of a transgene-based genetic sexing system for the silkworm, Bombyx mori. We developed a construct in which a positive feedback loop regulated by sex-specific alternative splicing leads to high-level expression of the tetracycline-repressible transactivator in females only. Transgenic animals show female-specific lethality during embryonic and early larval stages, leading to male-only cocoons. This transgene-based female-specific lethal system not only has wide application in sericulture, but also has great potential in lepidopteran pest control. PMID:23569267

  9. 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

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

    PubMed Central

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

    2014-01-01

    In the bivoltine strain of the silkworm, Bombyx mori, embryonic diapause is induced transgenerationally as a maternal effect. Progeny diapause is determined by the environmental temperature during embryonic development of the mother; however, its molecular mechanisms are largely unknown. Here, we show that the Bombyx TRPA1 ortholog (BmTrpA1) acts as a thermosensitive transient receptor potential (TRP) channel that is activated at temperatures above ∼21 °C and affects the induction of diapause in progeny. In addition, we show that embryonic RNAi of BmTrpA1 affects diapause hormone release during pupal-adult development. This study identifying a thermosensitive TRP channel that acts as a molecular switch for a relatively long-term predictive adaptive response by inducing an alternative phenotype to seasonal polyphenism is unique. PMID:24639527

  11. 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.

  12. Ecdysone 20-monooxygenase in eggs of the silkworm, Bombyx mori: enzymatic properties and developmental changes.

    PubMed

    Horike, N; Sonobe, H

    1999-01-01

    Ecdysone 20-monooxygenase in eggs of the silkworm Bombyx mori was characterized in relation to embryonic development. First, subcellular fractions were prepared by means of differential centrifugation, and analyzed using marker enzymes and antibodies against NADPH-cytochrome P450 reductase. It was demonstrated that most ecdysone 20-monooxygenase activity was associated with microsomes, and that there was little or no intrinsic mitochondrial ecdysone 20-monooxygenase. Next, conditions for the measurement of ecdysone 20-monooxygenase activity were established for the microsomal fraction, and changes in the enzyme activity were measured in diapause eggs and non-diapause eggs during early embryogenesis. It was demonstrated that enzyme activity in diapause eggs remained at a low level, while that in the non-diapause eggs increased from the gastrula stage. The increase in egg ecdysone 20-monooxygenase activity was prevented by actinomycin D and alpha-amanitin, suggesting that gene transcription is required for eliciting an increase in ecdysone 20-monooxygenase activity. PMID:10331236

  13. 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

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

    PubMed

    Walters, James R; Hardcastle, Thomas J

    2011-01-01

    Dosage compensation--equalizing gene expression levels in response to differences in gene dose or copy number--is classically considered to play a critical role in the evolution of heteromorphic sex chromosomes. As the X and Y diverge through degradation and gene loss on the Y (or the W in female-heterogametic ZW taxa), it is expected that dosage compensation will evolve to correct for sex-specific differences in gene dose. Although this is observed in some organisms, recent genome-wide expression studies in other taxa have revealed striking exceptions. In particular, reports that both birds and the silkworm moth (Bombyx mori) lack dosage compensation have spurred speculation that this is the rule for all female-heterogametic taxa. Here, we revisit the issue of dosage compensation in silkworm by replicating and extending the previous analysis. Contrary to previous reports, our efforts reveal a pattern typically associated with dosage compensated taxa: the global male:female expression ratio does not differ between the Z and autosomes. We believe the previous report of unequal male:female ratios on the Z reflects artifacts of microarray normalization in conjunction with not testing a major assumption that the male:female global expression ratio was unbiased for autosomal loci. However, we also find that the global Z chromosome expression is significantly reduced relative to autosomes, a pattern not expected in dosage compensated taxa. This combination of male:female parity with an overall reduction in expression for sex-linked loci is not consistent with the prevailing evolutionary theory of sex chromosome evolution and dosage compensation. PMID:21508430

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

    PubMed

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

    2015-04-01

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

  16. MicroRNA Let-7 regulates molting and metamorphosis in the silkworm, Bombyx mori.

    PubMed

    Ling, Lin; Ge, Xie; Li, Zhiqian; Zeng, Baosheng; Xu, Jun; Aslam, Abu F M; Song, Qisheng; Shang, Peng; Huang, Yongping; Tan, Anjiang

    2014-10-01

    MicroRNAs (miRNAs) are a class of endogenous, non-coding, regulatory RNA molecules that post-transcriptionally regulate gene expression by binding to the 3'UTRs of mRNA targets and thus cause their degradation or translational inhibition. In insects, important roles of miRNAs in various biological processes have been demonstrated in Drosophila melanogaster. However, biological roles of miRNAs are barely unveiled in the majority of insect species due to limited genetic tools. In the present study, we introduce the transgenic miRNA sponge (miR-SP) technology combining with the binary GAL4/UAS system in the domesticated silkworm, Bombyx mori, to exploit the biological function of an evolutionally conserved miRNA, let-7. We successfully established transgenic silkworm lines in which a miRNA sponge construct targeting BmLet-7 seed region was expressed in a ubiquitous manner directed by A3-GAL4 driver. Transgenic animals showed decreased expression of BmLet-7, leading to developmental arrestment during the larval-larval and larval-pupal transition. Simultaneously, expression levels of the predicted BmLet-7 target genes, FTZ-F1 and Eip74EF (E74), key regulatory factors in the ecdysone pathway, were elevated in transgenic animals. The current study is the first report on application of the transgenic miR-SP technology in non-drosophilid insects, which will not only contribute to better understanding of let-7 biological roles, but also greatly facilitate future miRNA functional analysis in insects. PMID:25016132

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

    PubMed

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

    2015-10-01

    In insects, cytochrome P450 monooxygenases (P450s) are involved in the metabolism of endogenous compounds such as steroid hormones and lipids. In this study, we measured the 20-hydroxyecdysone (20E)-induced transcriptional level of the CYP6ab4 gene using reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) with a dual spike-in strategy. We then probed possible physiological functions using RNAi experiments in the silkworm Bombyx mori. The activity of the CYP6ab4 promoter in various silkworm tissues was measured by firefly luciferase activity and normalized by Renilla luciferase activity. Our results showed that the activity of the CYP6ab4 promoter was highest in the malpighian tubule, followed by the fat body, the silk gland, the midgut, the epidermis, and the hemocyte. The essential region for basal and 20E-induced transcriptional activity was between -908 and -456 bp from the transcription start site. Through promoter truncation analysis using a dual-luciferase reporter assay in B. mori ovary cells (BmN), we showed that the region between -827 and -722 bp was essential for basal and 20E-induced transcriptional activity. Sequence analysis of this region revealed several potential transcriptional regulatory elements such as Hunchback (Hb) and BR-C Z. Mutation of the core bases of the BR-C Z binding site demonstrated that BR-C Z induces 20E-mediated CYP6ab4 transcription. Further identification of cis- and trans-elements and their roles in the upregulation of CYP6ab4 may be useful for elucidating the contribution of P450 to the response mechanism to 20E. PMID:25920486

  18. BmREEPa Is a Novel Gene that Facilitates BmNPV Entry into Silkworm Cells

    PubMed Central

    Wang, Wei; Pan, Cai-xia; Wu, Yun-fei; Du, Guo-yu; Chen, Peng; Lu, Cheng; Pan, Min-hui

    2015-01-01

    We previously established two silkworm cell lines, BmN-SWU1 and BmN-SWU2, from Bombyx mori ovaries. BmN-SWU1 cells are susceptible while BmN-SWU2 cells are highly resistant to BmNPV infection. Interestingly, we found that the entry of BmNPV into BmN-SWU2 cells was largely inhibited. To explore the mechanism of this inhibition, in this study we used isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative protein expression profiling and identified 629 differentially expressed proteins between the two cell lines. Among them, we identified a new membrane protein termed BmREEPa. The gene encoding BmREEPa transcribes two splice variants; a 573 bp long BmREEPa-L encoding a protein with 190 amino acids and a 501 bp long BmREEPa-S encoding a protein with 166 amino acids. BmREEPa contains a conserved TB2/DP, HVA22 domain and three transmembrane domains. It is localized in the plasma membrane with a cytoplasmic C-terminus and an extracellular N-terminus. We found that limiting the expression of BmREEPa in BmN-SWU1 cells inhibited BmNPV entry, whereas over-expression of BmREEPa in BmN-SWU2 cells promoted BmNPV entry. Our results also indicated that BmREEPa can interact with GP64, which is the key envelope fusion protein for BmNPV entry. Taken together, the findings of our study revealed that BmREEPa is required for BmNPV to gain entry into silkworm cells, and may provide insights for the identification of BmNPV receptors. PMID:26656276

  19. Two adenine nucleotide translocase paralogues involved in cell proliferation and spermatogenesis in the silkworm Bombyx mori.

    PubMed

    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

  20. Shotgun Proteomic Analysis on the Diapause and Non-Diapause Eggs of Domesticated Silkworm Bombyx mori

    PubMed Central

    Fan, Lanfen; Lin, Jianrong; Zhong, Yangsheng; Liu, Jingyi

    2013-01-01

    To clarify the molecular mechanisms of silkworm diapause, it is necessary to investigate the molecular basis at protein level. Here, the spectra of peptides digested from silkworm diapause and non-diapause eggs were obtained from liquid chromatography tandem mass spectrometry (LC-MS/MS) and were analyzed by bioinformatics methods. A total of 501 and 562 proteins were identified from the diapause and non-diapause eggs respectively, of which 309 proteins were shared commonly. Among these common-expressed proteins, three main storage proteins (vitellogenin precursor, egg-specific protein and low molecular lipoprotein 30 K precursor), nine heat shock proteins (HSP19.9, 20.1, 20.4, 20.8, 21.4, 23.7, 70, 90-kDa heat shock protein and heat shock cognate protein), 37 metabolic enzymes, 22 ribosomal proteins were identified. There were 192 and 253 unique proteins identified in the diapause and non-diapause eggs respectively, of which 24 and 48 had functional annotations, these unique proteins indicated that the metabolism, translation of the mRNA and synthesis of proteins were potentially more highly represented in the non-dipause eggs than that in the diapause eggs. The relative mRNA levels of four identified proteins in the two kinds of eggs were also compared using quantitative reverse transcription PCR (qRT-PCR) and showed some inconsistencies with protein expression. GO signatures of 486 out of the 502 and 545 out of the 562 proteins identified in the diapause and non-diapause eggs respectively were available. In addition, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed the Metabolism, Translation and Transcription pathway were potentially more active in the non-dipause eggs at this stage. PMID:23580252

  1. Melanin and urate act to prevent ultraviolet damage in the integument of the silkworm, Bombyx mori.

    PubMed

    Hu, Yong-Gang; Shen, Yi-Hong; Zhang, Ze; Shi, Gui-Qin

    2013-05-01

    The phenomenon that epidermal cells under the white stripes rather than black stripes contain many uric acid granules was found in larvae of several Lepidopteran species. However, the biological mechanism of this phenomenon is still unknown. In the present study, we take advantage of several silkworm (Bombyx mori) body color mutant strains to investigate the deposition patterns and biological mechanism of urate and melanin in the integuments of these mutant larvae. By imaging with transmission electron microscope, we found that there were some melanin granules in the larval cuticle in black body color mutant plain Black (p(B) ), but not in background strain plain (p) with white larval body color. In contrast, the larval epidermal cell of background strain had much more urate granules than that of black one. Furthermore, the uric acid content under the black stripes was significantly lower than that under the white stripes in a single individual of mottled stripe (p(S) ) with black and white stripes in each segment. Ultraviolet A (UVA) exposure experiments showed that the distinct oily (od) mutant individuals with translucent larval integument were more sensitive to the UVA damage than black body color mutant and background strain without any pigmentation in the larval cuticle. This is likely due to the absence of melanin granules and few urate granules in the integument of od mutant. Thus, both the deposited melanin granules in the cuticle and the abundant urate granules in the epidermis cells constitute effective barriers for the silkworm to resist UVA-induced damage. PMID:23575996

  2. Participation of d-serine in the development and reproduction of the silkworm Bombyx mori.

    PubMed

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

    2016-04-01

    The silkworm Bombyx mori contains high concentrations of free d-serine, an optical isomer of l-serine. To elucidate its function, we first investigated the localization of d-serine in various organs of silkworm larvae, pupae, and adult moths. Using immunohistochemical analysis with an anti-d-serine antibody, we found d-serine in the microvilli of midgut goblet and cylindrical cells and in peripheral matrix components of testicular and ovarian cells. By spectrophotometric analysis, d-serine was also found in the hemolymph and fat body. d-Alanine was not detected in the various organs by immunohistochemistry. Serine racemase, which catalyzes the inter-conversion of l- and d-serine, was found to co-localize with d-serine, and d-serine production from l-serine by intrinsic serine racemase was suggested. O-Phospho-l-serine is an inhibitor of serine racemase, and it was administered to the larvae to reduce the d-serine level. This reagent decreased the midgut caspase-3 level and caused a delay in spermatogenesis and oogenesis. The reagent also decreased mature sperm and egg numbers, suggesting d-serine participation in these processes. d-Serine administration induced an increase in pyruvate levels in testis, midgut, and fat body, indicating conversion of d-serine to pyruvate. On the basis of these results, together with our previous investigation of ATP biosynthesis in testis, we consider the possible involvement of d-serine in ATP synthesis for metamorphosis and reproduction. PMID:26828952

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

    PubMed Central

    Daubnerová, Ivana; Roller, Ladislav; Žitňan, Dušan

    2010-01-01

    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

  4. BmREEPa Is a Novel Gene that Facilitates BmNPV Entry into Silkworm Cells.

    PubMed

    Dong, Xiao-long; Liu, Tai-hang; Wang, Wei; Pan, Cai-xia; Wu, Yun-fei; Du, Guo-yu; Chen, Peng; Lu, Cheng; Pan, Min-hui

    2015-01-01

    We previously established two silkworm cell lines, BmN-SWU1 and BmN-SWU2, from Bombyx mori ovaries. BmN-SWU1 cells are susceptible while BmN-SWU2 cells are highly resistant to BmNPV infection. Interestingly, we found that the entry of BmNPV into BmN-SWU2 cells was largely inhibited. To explore the mechanism of this inhibition, in this study we used isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative protein expression profiling and identified 629 differentially expressed proteins between the two cell lines. Among them, we identified a new membrane protein termed BmREEPa. The gene encoding BmREEPa transcribes two splice variants; a 573 bp long BmREEPa-L encoding a protein with 190 amino acids and a 501 bp long BmREEPa-S encoding a protein with 166 amino acids. BmREEPa contains a conserved TB2/DP, HVA22 domain and three transmembrane domains. It is localized in the plasma membrane with a cytoplasmic C-terminus and an extracellular N-terminus. We found that limiting the expression of BmREEPa in BmN-SWU1 cells inhibited BmNPV entry, whereas over-expression of BmREEPa in BmN-SWU2 cells promoted BmNPV entry. Our results also indicated that BmREEPa can interact with GP64, which is the key envelope fusion protein for BmNPV entry. Taken together, the findings of our study revealed that BmREEPa is required for BmNPV to gain entry into silkworm cells, and may provide insights for the identification of BmNPV receptors. PMID:26656276

  5. Electrophysiological and Behavioral Responses of Male Fall Webworm Moths (Hyphantria cunea) to Herbivory-Induced Mulberry (Morus alba) Leaf Volatiles

    PubMed Central

    Tang, Rui; Zhang, Jin Ping; Zhang, Zhong Ning

    2012-01-01

    Volatile organic compounds (VOCs) were collected from damaged and intact mulberry leaves (Morus alba L., Moraceae) and from Hyphantria cunea larvae by headspace absorption with Super Q columns. We identified their constituents using gas chromatography-mass spectrometry, and evaluated the responses of male H. cunea antennae to the compounds using gas chromatography-flame ionization detection coupled with electroantennographic detection. Eleven VOC constituents were found to stimulate antennae of male H. cunea moths: ?-ocimene, hexanal, cis-3-hexenal, limonene, trans-2-hexenal, cyclohexanone, cis-2-penten-1-ol, 6-methyl-5-hepten-2-one, 4-hydroxy-4-methyl-2-pentanone, trans-3-hexen-1-ol, and 2,4-dimethyl-3-pentanol. Nine of these chemicals were released by intact, mechanically-damaged, and herbivore-damaged leaves, while cis-2-penten-1-ol was released only by intact and mechanically-damaged leaves and ?-ocimene was released only by herbivore-damaged leaves. Results from wind tunnel experiments conducted with volatile components indicated that male moths were significantly more attracted to herbivory-induced volatiles than the solvent control. Furthermore, male moths' attraction to a sex pheromone lure was increased by herbivory-induced compounds and ?-ocimene, but reduced by cis-2-penten-1-ol. A proof long-range field trapping experiment showed that the efficiency of sex pheromone lures in trapping male moths was increased by ?-ocimene and reduced by cis-2-penten-1-ol. PMID:23166622

  6. Structural analysis and anti-obesity effect of a pectic polysaccharide isolated from Korean mulberry fruit Oddi (Morus alba L.).

    PubMed

    Choi, Ji Won; Synytsya, Andriy; Capek, Peter; Bleha, Roman; Pohl, Radek; Park, Yong Il

    2016-08-01

    A water-soluble polysaccharide JS-MP-1 was isolated from Korean mulberry fruits Oddi (Morus alba L.). Sugar linkage analysis and NMR data confirmed that it is a rhamnogalacturonan type I (RG I) polymer carrying arabinan and arabinogalactan (AG II) side chains. JS-MP-1 reduced dose-dependently the viability of 3T3-L1 pre-adipocyte cells, significantly stimulated the cleavage of caspases 9 and 3 and poly (ADP-ribose) polymerase (PARP) and decreased the ratio of Bcl-2 to Bax expression level that led to mitochondrial dysfunction and apoptosis in pre-adipocyte cells. The apoptotic death was mediated by stimulation of MAPKs (ERK and p38) signalling pathway. These results suggest that JS-MP-1 is able to reduce the number of fat cells and the mass of adipose tissue via inhibition of pre-adipocyte proliferation and thus JS-MP-1 itself or a crude aqueous Oddi extract containing this polysaccharide can be used as functional ingredient of health-beneficial food supplements for the treatment or prevention of obesity disorders. PMID:27112865

  7. White Mulberry (Morus alba) Foliage Methanolic Extract Can Alleviate Aeromonas hydrophila Infection in African Catfish (Clarias gariepinus)

    PubMed Central

    Sheikhlar, Atefeh; Alimon, Abd Razk; Daud, Hassan; Saad, Chee R.; Webster, Carl D.; Meng, Goh Yong

    2014-01-01

    Two experiments were simultaneously conducted with Morus alba (white mulberry) foliage extract (MFE) as a growth promoter and treatment of Aeromonas hydrophila infection in separate 60 and 30 days trail (Experiments 1 and 2, resp.) in African catfish (Clarias gariepinus). In Experiment 1, four diets, control and control supplemented with 2, 5, or 7 g MFE/kg dry matter (DM) of diet, were used. In Experiment 2, fish were intraperitoneally infected with Aeromonas hydrophila and fed the same diets as experiment 1 plus additional two diets with or without antibiotic. Results of experiment 1 showed that growth was unaffected by dietary levels of MFE. Treatments with the inclusion of MFE at the levels of 5 and 7 g/Kg DM had no mortality. Red blood cells (RBC), albumin, and total protein were all higher for the treatments fed MFE (5 and 7 g/Kg DM). Results of experiment 2 showed RBC, hemoglobin, hematocrit, globulin, albumin, and total protein improved with the increase in MFE in the infected fish. The dietary MFE at the level of 7 g/kg DM reduced mortality rate. In conclusion, MFE at the level of 7 g/kg DM could be a valuable dietary supplement to cure the infected fish. PMID:25574488

  8. Preparation of the Branch Bark Ethanol Extract in Mulberry Morus alba, Its Antioxidation, and Antihyperglycemic Activity In Vivo

    PubMed Central

    Wang, Shu; Fang, Meng; Ma, Yong-Lei

    2014-01-01

    The biological activities of the branch bark ethanol extract (BBEE) in the mulberry Morus alba L. were investigated. The determination of active component showed that the flavonoids, phenols, and saccharides are the major components of the ethanol extract. The BBEE had a good scavenging activity of the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical with around 100 μg/mL of IC50 value. In vitro assay revealed that the BBEE strongly inhibited both α-glucosidase and sucrase activities whose IC50 values were 8.0 and 0.24 μg/mL, respectively. The kinetic analysis showed that the BBEE as a kind of α-glucosidase inhibitor characterized a competitive inhibition activity. Furthermore, the carbohydrate tolerance of the normal mice was obviously enhanced at 0.5 h (P < 0.05) and 1.0 h (P < 0.05) after the BBEE intragastric administration as compared to negative control. At 0.5, 1.0, 1.5, and 2.0 h after the intragastric administration with starch, the postprandial hyperglycemia of the type 2 diabetic mice can be significantly decreased (P < 0.01) by supplying various concentrations of the BBEE (10–40 mg/kg body weight). Therefore, the BBEE could effectively inhibit the postprandial hyperglycemia as a novel α-glucosidase activity inhibitor for the diabetic therapy. PMID:24587809

  9. Proximate Composition and Antioxidant Potential of Leaves from Three Varieties of Mulberry (Morus sp.): A Comparative Study

    PubMed Central

    Iqbal, Shahid; Younas, Umer; Sirajuddin; Chan, Kim Wei; Sarfraz, Raja Adil; Uddin, Kamal

    2012-01-01

    In this study, leaves of three indigenous varieties of Mulberry namely, Morus alba L., Morus nigra L. and Morus rubra L. were investigated for their antioxidant potential and their proximate composition was determined. The yields of 80% methanolic extracts ranged between 8.28–13.89%. The contents of total phenolics (TPC), total flavonoids (TFC) and ascorbic acid (AA) ranged between 16.21–24.37 mg gallic acid equivalent (GAE)/g, 26.41–31.28 mg rutin equivalent (RE)/g and 0.97–1.49 mg/g, respectively. The antioxidant activity of leaf extracts was evaluated by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH•) radical scavenging actity, 2,2′-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid (ABTS•+) radical cation scavenging capacity and ferric ion reducing power and values ranged between 1.89–2.12, 6.12–9.89 and 0.56–0.97 mM Trolox equivalent/g of dried leaves, respectively. The investigated features reveal good nutritive and antioxidant attributes of all the varieties with mutually significant differences. PMID:22837655

  10. Wild 2 Close Look

    NASA Technical Reports Server (NTRS)

    2004-01-01

    [figure removed for brevity, see original site] Figure 1

    This image shows the comet Wild 2, which NASA's Stardust spacecraft flew by on Jan. 2, 2004. This image is the closest short exposure of the comet, taken at an11.4-degree phase angle, the angle between the camera, comet and the Sun. The listed names on the diagram (see Figure 1) are those used by the Stardust team to identify features. 'Basin' does not imply an impact origin.

  11. Wild 2 Features

    NASA Technical Reports Server (NTRS)

    2004-01-01

    [figure removed for brevity, see original site] Figure 1

    These images taken by NASA's Stardust spacecraft highlight the diverse features that make up the surface of comet Wild 2. Side A (see Figure 1) shows a variety of small pinnacles and mesas seen on the limb of the comet. Side B (see Figure 1) shows the location of a 2-kilometer (1.2-mile) series of aligned scarps, or cliffs, that are best seen in the stereo images.

  12. Wild Duck Cluster

    NASA Technical Reports Server (NTRS)

    2005-01-01

    On April 7, 2005, the Deep Impact spacecraft's Impactor Target Sensor camera recorded this image of M11, the Wild Duck cluster, a galactic open cluster located 6 thousand light years away. The camera is located on the impactor spacecraft, which will image comet Tempel 1 beginning 22 hours before impact until about 2 seconds before impact. Impact with comet Tempel 1 is planned for July 4, 2005.

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

    NASA Astrophysics Data System (ADS)

    Zhang, Zong-jian; Li, Jia; Sun, Fu-sheng; Dickon, H. L. Ng; Luen Kwong, Fung

    2010-06-01

    A simple and green technique has been developed to prepare hierarchical biomorphic ZrO2-CeO2, using silkworm silk as the template. Different from traditional immersion technics, the whole synthesis process depends more on the restriction or direction functions of the silkworm silk template. The analytic results showed that ZrO2-CeO2 exhibited a well-crystallized hierarchically interwoven hollow fiber structure with 16-28 μm in diameter. The grain size of the sample calcined at 800 °C was about 14 nm. Consequently, the interwoven meshwork at three dimensions is formed due to the direction of biotemplate. The action mechanism is summarily discussed here. It may bring the biomorphic ZrO2-CeO2 nanomaterials with hierarchical interwoven structures to more applications, such as catalysts.

  14. 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

  15. The 5'-UTR intron of the midgut-specific BmAPN4 gene affects the level and location of expression in transgenic silkworms.

    PubMed

    Jiang, Liang; Huang, Chunlin; Sun, Qiang; Guo, Huizhen; Cheng, Tingcai; Peng, Zhengwen; Dang, Yinghui; Liu, Weiqiang; Xu, Guowen; Xia, Qingyou

    2015-08-01

    Introns are important for regulating gene expression. BmAPN4, which has a 5'-UTR upstream intron (5 UI), is specifically expressed in the entire silkworm midgut. In our previous study, the promoter region upstream of the 5 UI of BmAPN4 was cloned and identified as the P3 promoter (P3P) with activity only in the anterior midgut. In this study, the sequence consisting of the P3P and the 5 UI was cloned and named as P3P+5 UI. A transgenic vector was constructed in which EGFP was controlled by P3P+5 UI. Transgenic P3+5 UI silkworms were generated by embryo microinjection. RT-PCR showed P3P+5 UI activity throughout the larval stage. Intense green fluorescence was seen only in the entire midgut of P3+5 UI silkworms and expression was confirmed by RT-PCR. qPCR revealed that expression of EGFP in the anterior midgut of P3+5 UI silkworms was 64% higher than in P3 silkworms, indicating the 5 UI sustained intron-mediated enhancement of gene expression. These results suggested that the BmAPN4 5 UI affected the level and site of expression. The 5 UI was cloned and added behind P2P, another specific promoter with activity only in the anterior midgut of silkworm, to construct the P2P+5 UI and transgenic P2+5 UI silkworms. Expression patterns were the same for P2P+5 UI and P2P, suggesting that the 5UI of BmAPN4 did not affect P2P. This study found that the BmAPN4 5 UI affected the amount and location of gene expression. Its influence appeared to be dependent on a specific promoter. PMID:25982022

  16. 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

  17. Ecotoxicology of Wild Mammals

    USGS Publications Warehouse

    2001-01-01

    An international group of 32 scientists has critically reviewed the scientific literature on exposure and effects of environmental contaminants in wild mammals. The underlying theme of this text is encompassed by the following four questions: What exactly do we know about environmental contaminants in mammals? What are the commonalities and differences between mammal orders/species in the effects that contaminants have? How and to what degree of accuracy can we predict the adverse effects of environmental contaminants on mammalian wildlife? How significant are contaminant insults compared with other density-independent and -dependent factors such as habitat loss, climatic factors and disease? The book is organized three topical sections including introductory chapters that provide a background on environmental contaminants and the mammalian orders, eight taxonomic chapters discussing all aspects of the exposure to and effects of contaminants in mammalian orders, and four thematic chapters that review and discuss generic issues including biomarkers, prediction and extrapolation of exposure and effects, hazard and risk assessment, and the relative significance of contaminants on mammals compared with other commonly encountered stressors. A final a summary chapter identifies phylogenetic trends, critical data gaps, and overarching research needs. Although the absolute number of toxicological studies in domesticated and wild mammals eclipses that wildlife species, a detailed examination of our knowledge base reveals that information for 'wild' birds is actually greater than that for 'wild' mammals. Of the various mammalian taxa, ecotoxicological data is most noticeably lacking for marsupials and monotremes. In contrast, rodents (comprising 43% of all mammal species) have been studied extensively, despite evidence of their tolerance to some organochlorine compounds, rodenticides, and even radionuclides. Mammalian species at greatest risk of exposure include those that consume a high percentage of their body weight on a daily basis. Aquatic mammals tend to bioaccumulate tremendous burdens of lipophilic contaminants, although storage in their fat depots may actually limit toxicity. Carnivores appear to be more sensitive to adverse effects of environmental contaminants than herbivores. Remarkably few of the thousands of compounds manufactured worldwide have been toxicologically evaluated in wild mammals, and concentrations of even fewer have been monitored in tissues.

  18. De Novo Assembly of Expressed Transcripts and Global Transcriptomic Analysis from Seedlings of the Paper Mulberry (Broussonetia kazinoki x Broussonetia papyifera)

    PubMed Central

    Xianjun, Peng; Linhong, Teng; Xiaoman, Wang; Yucheng, Wang; Shihua, Shen

    2014-01-01

    The paper mulberry is one of the multifunctional tree species in agroforestry systems and is also commonly utilized in traditional medicine in China and other Asian countries. However, little is known about its molecular genetics, which hinders research on and exploitation of this valuable resource. To discern the correlation between gene expression and the essential properties of the paper mulberry, we performed a transcriptomics analysis, assembling a total of 37,725 unigenes from 54,638,676 reads generated by RNA-seq. Among these, 22,692 unigenes showed greater than 60% similarity with genes from other species. The lengths of 13,566 annotated unigenes were longer than 1,000 bp. Functional clustering analysis with COG (Cluster of Orthologous Groups) revealed that 17,184 unigenes are primarily involved in transcription, translation, signal transduction, carbohydrate metabolism, secondary metabolism, and energy metabolism. GO (Gene Ontology) annotation suggests enrichment of genes encoding antioxidant activity, transporter activity, biosynthesis, metabolism and stress response, with a total of 30,659 unigenes falling in these categories. KEGG (Kyoto Encyclopedia of Genes and Genomes) metabolic pathway analysis showed that 7,199 unigenes are associated with 119 metabolic pathways. In addition to the basic metabolism, these genes are enriched for plant pathogen interaction, flavonoid metabolism and other secondary metabolic processes. Furthermore, differences in the transcriptomes of leaf, stem and root tissues were analyzed and 7,233 specifically expressed unigenes were identified. This global expression analysis provided novel insights about the molecular mechanisms of the biosynthesis of flavonoid, lignin and cellulose, as well as on the response to biotic and abiotic stresses including the remediation of contaminated soil by the paper mulberry. PMID:24848504

  19. Combined Mulberry Leaf and Fruit Extract Improved Early Stage of Cutaneous Wound Healing in High-Fat Diet-Induced Obese Mice.

    PubMed

    Eo, Hyeyoon; Lim, Yunsook

    2016-02-01

    Obesity is a pandemic that causes many health challenges, including difficulties in achieving proper wound healing without complications. The current study investigated the role of NLRP3 inflammasome in the early stages of cutaneous wound healing and the effect of combined mulberry leaf and fruit extract (MLFE) on cutaneous NLRP inflammasome involvement in delayed wound healing mice with high-fat diet (HFD)-induced obesity. After obesity was induced by HFD for 10 weeks, the mice were supplemented with MLFE (at a dose of 500 mg/kg containing 333.3 mg/kg of mulberry leaf extract and 166.7 mg/kg of mulberry fruit extract) by gavage, 5 days/week for 12 weeks. MLFE supplementation ameliorated delayed wound closure in obese mice. While wound size was positively correlated with fasting blood glucose level during the early stage of wound healing, it was strongly correlated with body weight gain and body fat mass during the later stage of wound healing. Under obese conditions, the levels of NLRP3 inflammasome and its related markers (pro-caspase-1 and precursor/mature interleukin 1 beta) were increased at a basal level, but the NLRP3 inflammasome was suppressed during the inflammatory stage of cutaneous wound healing. However, MLFE supplementation stimulated cutaneous NLRP3 inflammasome in HFD-induced obese mice (day 3). Taken together, stimulating the NLRP3 inflammasome might be beneficial in the early inflammatory stage of cutaneous wound healing and MLFE could be a potential therapeutic intervention in delayed wound healing through activation of the NLRP3 inflammasome in obesity. PMID:26491791

  20. A mixture of extracts of black and green teas and mulberry leaf did not reduce weight gain in rats fed a high-fat diet.

    PubMed

    Fallon, Elizabeth; Zhong, Litao; Furne, Julie K; Levitt, Michael

    2008-03-01

    Tea extracts are used in many over-the-counter preparations claiming to promote weight loss. The rationale for this usage includes reports that green tea extract increases thermogenesis, and extracts of green and black tea and mulberry leaf inhibit the digestion/absorption of carbohydrate and fat. The investigators in this study tested the potential of increasing doses of a mixture of three extracts (50-percent black tea, 20-percent green tea, and 30-percent mulberry) to induce weight loss, steatorrhea, and blood lipid alterations in rats ingesting a high-fat diet, ad lib. The mixture was incorporated into chow in quantities of 0.5-, 3.0-, and 6.0 percent by weight; a control group received only chow. Food intake and weight were monitored daily, and quantitative fecal fat measurements were obtained weekly for four weeks. The 3.0- and 6.0-percent chows significantly increased fecal fat excretion to 15 percent of dietary fat intake (controls: 5 percent); however, no significant reduction in weight gain was observed. After four weeks of treatment, the 3.0- and 6.0-percent dosages were associated with significant reductions in serum triglycerides and increases in high-density lipoprotein (HDL) cholesterol. However, these chow concentrations were associated with significant increases in serum ALT, and the 6.0-percent chow markedly increased serum alkaline phosphatase. This study does not provide support for the utility of this combination of black tea, green tea, and mulberry extracts in weight-loss regimens and indicates that high doses of this extract combination may be hepatotoxic. PMID:18377102

  1. Linkage and mapping analyses of the normal marking gene +P in the silkworm (Bombyx mori) using SSR markers.

    PubMed

    Wei, G Q; Yu, L; Liu, C L; Zhu, B J; Ding, H J

    2013-01-01

    In the silkworm, Bombyx mori, normal markings are mainly controlled by the +P gene, which is located on the second chromosome. Due to a lack of crossing over in females, reciprocal backcrossed F1 (BC1) progenies were used for linkage analysis and mapping of the +P gene based on an SSR linkage map using silkworm strains P50 and H9, which are normal marking and sex-limited marking, respectively. The +P gene was found to be linked to 3 SSR markers. Using a reciprocal BC1M cross, we constructed a linkage map of 22.5 cM, with +P mapped at 11.3 cM and the nearest SSR marker S0206 at a distance of 3.0 cM. Based on a fine genome map of domesticated silkworms, Kaikoblast analysis showed that the physical distance between the nearest markers (containing the +P gene) is 995 kb. Further analysis showed that BGIBMGA009689, BGIBMGA009688, and BGIBMGA009687 are closer to +P, and that BGIBMGA009689 is closest to +P, with a physical distance of 19.1 kb. PMID:23359060

  2. A genomewide survey of homeobox genes and identification of novel structure of the Hox cluster in the silkworm, Bombyx mori.

    PubMed

    Chai, Chun-Li; Zhang, Ze; Huang, Fei-Fei; Wang, Xian-Yan; Yu, Quan-You; Liu, Bin-Bin; Tian, Tian; Xia, Qing-You; Lu, Cheng; Xiang, Zhong-Huai

    2008-12-01

    Homeobox genes encode transcriptional factors that play crucial roles in a variety of developmental pathways from unicellular to multicellular eukaryotes. We have identified 102 homeobox genes in the typical insect of Lepidoptera, Bombyx mori, based on the newly assembled genome sequence with 9X coverage. These identified homeobox genes were categorized into nine classes including at least 74 families. The available ESTs and microarray data at present confirmed that more than half of them were expressed during silkworm developmental processes. Orthologs of pb, zen and ftz were newly identified in the Bombyx Hox cluster on chromosome 6. Interestingly, a special group of 12 tandemly duplicated homeobox genes was found located between Bmpb and Bmzen in the Bombyx Hox cluster, suggesting that Hox cluster might have experienced a lineage-specific expansion in the silkworm. A detailed analysis on genome data reveals that a split exists between Bmlab and Bmpb. Our data provide valuable information for future research on the development and evolution of silkworm. PMID:19280701

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

    PubMed

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

    2016-05-01

    Cuticle is mainly made of chitin filaments embedded in a matrix of cuticular proteins (CPs). Cuticular chitins have minor differences, whereas CPs are widely variable with respect to their sequences and structures. To understand the molecular basis underlying the mechanical properties of cuticle, it is necessary to know which CPs interact with chitin and how they are assembled into the cuticle structure. In the present study, a chitin-binding assay was performed followed by liquid chromatography-tandem mass spectrometry to identify the extracted proteins from the larval cuticle of silkworm, Bombyx mori. There were 463 proteins identified from the silkworm larval cuticle, 200 of which were recovered in the chitin-binding fraction. A total of 103 proteins were annotated as CPs, which were classified into 11 CP families based on their conserved motifs, including CPR, CPAP, CPT, CPF and CPFL, CPCFC, chitin_bind 3, BmCPH2 homologues, BmCPH9 homologues, BmCPG1 homologues, BmCPG20 homologues, and BmCPG21 homologues. A total of five CP families were newly identified in the chitin-binding fraction, thereby providing new information and insight into the composition, structure, and function of the silkworm larval cuticle. PMID:26972338

  4. INVOLVEMENT OF PEPTIDOGLYCAN RECOGNITION PROTEIN L6 IN ACTIVATION OF IMMUNE DEFICIENCY PATHWAY IN THE IMMUNE RESPONSIVE SILKWORM CELLS.

    PubMed

    Tanaka, Hiromitsu; Sagisaka, Aki

    2016-06-01

    The immune deficiency (Imd) signaling pathway is activated by Gram-negative bacteria for producing antimicrobial peptides (AMPs). In Drosophila melanogaster, the activation of this pathway is initiated by the recognition of Gram-negative bacteria by peptidoglycan (PGN) recognition proteins (PGRPs), PGRP-LC and PGRP-LE. In this study, we found that the Imd pathway is involved in enhancing the promoter activity of AMP gene in response to Gram-negative bacteria or diaminopimelic (DAP) type PGNs derived from Gram-negative bacteria in an immune responsive silkworm cell line, Bm-NIAS-aff3. Using gene knockdown experiments, we further demonstrated that silkworm PGRP L6 (BmPGRP-L6) is involved in the activation of E. coli or E. coli-PGN mediated AMP promoter activation. Domain analysis revealed that BmPGRP-L6 contained a conserved PGRP domain, transmembrane domain, and RIP homotypic interaction motif like motif but lacked signal peptide sequences. BmPGRP-L6 overexpression enhances AMP promoter activity through the Imd pathway. BmPGRP-L6 binds to DAP-type PGNs, although it also binds to lysine-type PGNs that activate another immune signal pathway, the Toll pathway in Drosophila. These results indicate that BmPGRP-L6 is a key PGRP for activating the Imd pathway in immune responsive silkworm cells. PMID:26991439

  5. Quantitative proteomic and transcriptomic analyses of molecular mechanisms associated with low silk production in silkworm Bombyx mori.

    PubMed

    Wang, Shao-Hua; You, Zheng-Ying; Ye, Lu-Peng; Che, Jiaqian; Qian, Qiujie; Nanjo, Yohei; Komatsu, Setsuko; Zhong, Bo-Xiong

    2014-02-01

    To investigate the molecular mechanisms underlying the low fibroin production of the ZB silkworm strain, we used both SDS-PAGE-based and gel-free-based proteomic techniques and transcriptomic sequencing technique. Combining the data from two different proteomic techniques was preferable in the characterization of the differences between the ZB silkworm strain and the original Lan10 silkworm strain. The correlation analysis showed that the individual protein and transcript were not corresponded well, however, the differentially changed proteins and transcripts showed similar regulated direction in function at the pathway level. In the ZB strain, numerous ribosomal proteins and transcripts were down-regulated, along with the transcripts of translational related elongation factors and genes of important components of fibroin. The proteasome pathway was significantly enhanced in the ZB strain, indicating that protein degradation began on the third day of fifth instar when fibroin would have been produced in the Lan10 strain normally and plentifully. From proteome and transcriptome levels of the ZB strain, the energy-metabolism-related pathways, oxidative phosphorylation, glycolysis/gluconeogenesis, and citrate cycle were enhanced, suggesting that the energy metabolism was vigorous in the ZB strain, while the silk production was low. This may due to the inefficient energy employment in fibroin synthesis in the ZB strain. These results suggest that the reason for the decreasing of the silk production might be related to the decreased ability of fibroin synthesis, the degradation of proteins, and the inefficiency of the energy exploiting. PMID:24428189

  6. Comparative proteomics analysis of silkworm hemolymph during the stages of metamorphosis via liquid chromatography and mass spectrometry.

    PubMed

    Hou, Yong; Zhang, Yan; Gong, Jing; Tian, Sha; Li, Jianwei; Dong, Zhaoming; Guo, Chao; Peng, Li; Zhao, Ping; Xia, Qingyou

    2016-05-01

    The silkworm is a lepidopteran insect that has an open circulatory system with hemolymph consisting of blood and lymph fluid. Hemolymph is not only considered as a depository of nutrients and energy, but it also plays a key role in substance transportation, immunity response, and proteolysis. In this study, we used LC-MS/MS to analyze the hemolymph proteins of four developmental stages during metamorphosis. A total of 728 proteins were identified from the hemolymph of the second day of wandering stage, first day of pupation, ninth day of pupation, and first day as an adult moth. GO annotations and categories showed that silkworm hemolymph proteins were enriched in carbohydrate metabolism, proteolysis, protein binding, and antibacterial humoral response. The levels of nutrient, immunity-related, and structural proteins changed significantly during development and metamorphosis. Some, such as cuticle, odorant-binding, and chemosensory proteins, showed stage-specific expression in the hemolymph. In addition, the expression of several antimicrobial peptides exhibited their highest level of abundance in the hemolymph of the early pupal stage. These findings provide a comprehensive proteomic insight of the silkworm hemolymph and suggest additional molecular targets for studying insect metamorphosis. PMID:26960012

  7. Identification of a microspordium isolated from Megacopta cribraria (Hemiptera: Plataspidae) and characterization of its pathogenicity in silkworms.

    PubMed

    Xing, Dongxu; Li, Li; Liao, Sentai; Luo, Guoqing; Li, Qingrong; Xiao, Yang; Dai, Fanwei; Yang, Qiong

    2014-11-01

    A new microsporidium isolated from Megacopta cribraria was characterized by both biological characteristics and phylogenetic analysis. Moreover, its pathogenicity to silkworms was also studied. The spores are oval in shape and measured 3.64 ± 0.2 × 2.20 ± 0.2 μm in size. Its ultrastructure is characteristic of the genus Nosema: a diplokaryon, 13-14 polar filament coils and posterior vacuole. Its life cycle includes meronts, sporonts, sporoblasts and mature spores, with a typical diplokaryon in each stage and propagation in a binary fission. A phylogenetic tree based on SSU rRNA and rRNA ITS gene sequence analysis further indicated that the parasite is closely related to Nosema bombycis and should be placed in the genus Nosema and sub-group 'true' Nosema. Furthermore, the microsporidium heavily infects lepidopteran silkworm insect and can be transmitted per os (horizontally) and transovarially (vertically). Our findings showed that the microsporidium belongs to the 'true' Nosema group within the genus Nosema and heavily infects silkworms. Based on the information obtained during this study, we named this new microsporidium isolated from M. cribraria as Nosema sp. MC. PMID:25173855

  8. A novel method to fabricate superhydrophobic surfaces based on well-defined mulberry-like particles and self-assembly of polydimethylsiloxane

    NASA Astrophysics Data System (ADS)

    Yang, Jinxin; Pi, Pihui; Wen, Xiufang; Zheng, Dafeng; Xu, Mengyi; Cheng, Jiang; Yang, Zhuoru

    2009-01-01

    A superhydrophobic surface was obtained by combining application of CaCO 3/SiO 2 mulberry-like composite particles, which originated from violent stirring and surface modification, and self-assembly of polydimethylsiloxane. Water contact angle and sliding angle of the superhydrophobic surface were measured to be about 164 ± 2.5° and 5°, respectively. The excellent hydrophobicity is attributed to the synergistic effect of micro-submicro-nano-meter scale roughness (fabricated by composite particles) and the low surface energy (provided by polydimethylsiloxane). This procedure makes it possible for widespread applications of superhydrophobic film due to its simplicity and practicability.

  9. Antibiotic resistance in wild birds

    PubMed Central

    Bonnedahl, Jonas

    2014-01-01

    Wild birds have been postulated as sentinels, reservoirs, and potential spreaders of antibiotic resistance. Antibiotic-resistant bacteria have been isolated from a multitude of wild bird species. Several studies strongly indicate transmission of resistant bacteria from human rest products to wild birds. There is evidence suggesting that wild birds can spread resistant bacteria through migration and that resistant bacteria can be transmitted from birds to humans and vice versa. Through further studies of the spatial and temporal distribution of resistant bacteria in wild birds, we can better assess their role and thereby help to mitigate the increasing global problem of antibiotic resistance. PMID:24697355

  10. Teaching in wild meerkats.

    PubMed

    Thornton, Alex; McAuliffe, Katherine

    2006-07-14

    Despite the obvious benefits of directed mechanisms that facilitate the efficient transfer of skills, there is little critical evidence for teaching in nonhuman animals. Using observational and experimental data, we show that wild meerkats (Suricata suricatta) teach pups prey-handling skills by providing them with opportunities to interact with live prey. In response to changing pup begging calls, helpers alter their prey-provisioning methods as pups grow older, thus accelerating learning without the use of complex cognition. The lack of evidence for teaching in species other than humans may reflect problems in producing unequivocal support for the occurrence of teaching, rather than the absence of teaching. PMID:16840701

  11. Operation Wild Dog.

    PubMed

    Poole, P

    'Operation Wild Dog' was the title given to an incredible escapade by two Army Dental Corps soldiers who decided to mount their own two-man raid on occupied France in 1942. In an attempt to forestall claims of desertion, they involved Prime Minister Winston Churchill and, when they ultimately faced a court martial, were lauded in the press for their daredevil exploits. Inspired by raids carried out by Special Services units on Norway, Italy and France, Sergeant Peter King, a regular soldier and dental clerk orderly, and Private Thomas Leslie Cuthbertson, a trainee dental mechanic, set about their unofficial raid, outlined below. PMID:1467013

  12. Selection of reference genes for analysis of stress-responsive genes after challenge with viruses and temperature changes in the silkworm Bombyx mori.

    PubMed

    Guo, Huizhen; Jiang, Liang; Xia, Qingyou

    2016-04-01

    Viruses and high temperature (HT) are the primary threats to silkworms. Changes in the expression of stress-response genes can be measured using quantitative polymerase chain reaction (qPCR) after exposure to viruses or HT. However, appropriate reference genes (RGs) for qPCR data normalization have not been established in this organism. In this study, we summarized the RGs used in the previous silkworm studies after infection with Bombyx mori nucleopolyhedrovirus (BmNPV), B. mori cytoplasmic polyhedrosis virus (BmCPV), or B. mori densovirus (BmDNV) or after HT treatment. The expression levels of these RGs were extracted from silkworm transcriptome data to screen for candidate RGs that were unaffected by the experimental conditions. Actin-1 (A1), actin-3 (A3), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation initiation factor 4a (TIF-4A) were selected for further qPCR verification. The results of RNA-seq and qPCR showed that GAPDH and TIF-4A were suitable RGs after BmNPV challenge or HT stress, whereas TIF-4A was an appropriate RG for BmCPV or BmDNV-Z challenge in silkworms. These results suggested that TIF-4A may be the most appropriate RG for gene expression analysis after challenge with viruses or HT in silkworms. PMID:26437927

  13. Species-specific expansion of C2H2 zinc-finger genes and their expression profiles in silkworm, Bombyx mori.

    PubMed

    Duan, Jun; Xia, Qingyou; Cheng, Daojun; Zha, Xingfu; Zhao, Ping; Xiang, Zhonghuai

    2008-12-01

    Most C2H2 zinc-finger proteins (ZFPs) function as sequence-specific DNA-binding transcription factors, and play important roles in a variety of biology processes, such as development, differentiation, and tumor suppression. By searching the silkworm genome with a HMM model of C2H2 zinc-fingers, we have identified a total of 338 C2H2 ZFPs. Most of the ZFP genes were clustered on chromosomes and showed uneven distribution in the genome. Over one third of genes were concentrated on chromosome 11, 15 and 24. Phylogenetic analysis classified all silkworm C2H2 ZFPs into 75 families; 63 of which belong to evolutionarily conserved families. In addition, 188 C2H2 ZFP genes (55.6%) are species-specific to the silkworm. A species-specific expansion of a family with 39 members in a tandem array on chromosome 24 may explain the higher number of species-specific ZFPs in silkworm compared to other organisms. The expression patterns of C2H2 ZFP genes were also examined by microarray analysis. Most of these genes were actively expressed among different tissues on day 3 of the fifth instar. The results provide insight into the biological functions of the silkworm C2H2 ZFP genes in metamorphism and development. PMID:18835444

  14. Research on the Natural Enemies of the Mulberry Scale, Pseudaulacaspis pentagona (Targioni), in Tea Fields in Shizuoka Prefecture, Japan

    NASA Astrophysics Data System (ADS)

    Ozawa, Akihito; Kubota, Sakae; Kaneko, Shuji; Ishigami, Shigeru

    Species composition and the seasonal prevalence of natural enemies on the mulberry scale Pseudaulacaspis pentagona (Targioni) in tea fields in Shizuoka Prefecture of Japan were investigated by monitoring methods using yellow sticky traps hung on the branches under leaf layers. The species captured by the sticky traps were as follows: 5 species of parasitic wasps,Arrhenophagus albitibiae Girault, Pteroptrix orientalis (Silvestri), Thomsonisca indica Hayat (this species was identified as Thomsonisca amathus in Japan), and Epitetracnemus comis Noyes & Ren; 1 species of hyperparasites, Marietta carnesi (Howard); and 3 species of coleopteran predators, Pseudoscymnus hareja Weise, Chilocorus kuwanae Silvestri, and Cybocephalus nipponicus Endrody-Younga. Further, 1 Cecidomyiidae species (predatory gall midge), namely,Dentifibula sp., was captured by sticky traps. Among the parasitoids captured, A. albitibiae was the most abundant species, followed by P. orientalis. Among the predacious beetles captured, P. hareja was the dominant species. A. albitibiae demonstrated 5 or 6 peaks of seasonal prevalence in a year, and P. orientalis and T. indica exhibited 3 peaks of seasonal prevalence in a year. P. hareja and Dentifibula sp. demonstrated 3 indistinct peaks of seasonal prevalence in a year. The peak dates of A. albitibiae, P. orientalis, T. indica, and Dentifibula sp. were compared with those of the first instar larvae and adult males of the hosts, P. pentagona, which were captured by sticky traps. The relationships between the total numbers of each generation captured by sticky traps of the parasitoids A. albitibiae and the host P. pentagona over a period of 2 years revealed similar changes in the dynamics of the host-parasitoid models of Nicholson and Bailey (1935).This suggested that A. albitibiae was one of the most important natural enemies against P.pentagona in tea fields.

  15. Research on the Natural Enemies of the Mulberry Scale, Pseudaulacaspis pentagona (Targioni), in Tea Fields in Shizuoka Prefecture, Japan

    NASA Astrophysics Data System (ADS)

    Ozawa, Akihito; Kubota, Sakae; Kaneko, Shuji; Ishigami, Shigeru

    The mulberry scale Pseudaulacaspis pentagona (Targioni) is one of the most important pests on tea trees in Japan; in particular, severe outbreaks have occurred in Shizuoka Prefecture in recent years. Natural enemies of the scale are considered to be one of important factors for controlling the scale population, and it is necessary to clarify the actual condition of the natural enemies of the scale in tea fields. We investigated the species and species composition of natural enemies of the scale in tea fields in Shizuoka Prefecture of Japan in 2002 and 2003 by identifying the parasitoids that emerged and dissecting the adult female scales. We identified 4 species of primary parasites, namely, Arrhenophagus albitibiae Girault, Pteroptrix orientalis (Silvestri), Thomsonisca indica? Hayat (this species was identified as T. amathus in Japan) and Epitetracnemus comis Noyes & Ren, and 2 species of hyperparasites, namely, Marietta carnesi (Howard) and Zaomma near lambinus (Walker). We also identified the following 3 species of coleopteran predators: Pseudoscymnus hareja Weise, Chilocorus kuwanae Silvestri, and Cybocephalus nipponicus Endrody-Younga. Further, 1 species of Cecidomyiidae (predatory gall midge), namely, Dentifibula sp. was confirmed. The primary dominant parasitoid and predatory beetle species were A. albitibiae and P. hareja, respectively. The species composition of the parasitoids that emerged changed with location and generation of the scale, and A. albitibiae was the major species in the overwintering generation of the scale. However, from the first to the second generation, the frequency of species other than A. albitibiae increased, i.e., species diversity increased. The percentage parasitism of all of the parasitoids increased with the alternation of scale generations, and there were tea fields in which the predatory Cecidomyiidae Dentifibula sp. became the primary dominant species at the second generation of the scale. Therefore, it was suggested that interspecific competition and intraguild predation occurred among the natural enemies.

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

    PubMed

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

    2005-08-01

    In the silkworm, Bombyx mori (female, ZW; male, ZZ), femaleness is determined by the presence of a single W chromosome, irrespective of the number of autosomes or Z chromosomes. The W chromosome is devoid of functional genes, except the putative female-determining gene (Fem). However, there are strains in which chromosomal fragments containing autosomal markers have been translocated on to W. In this study, we analysed the W chromosomal regions of the Zebra-W strain (T(W;3)Ze chromosome) and the Black-egg-W strain (T(W;10)+(w-2) chromosome) at the molecular level. Initially, we undertook a project to identify W-specific RAPD markers, in addition to the three already established W-specific RAPD markers (W-Kabuki, W-Samurai and W-Kamikaze). Following the screening of 3648 arbitrary 10-mer primers, we obtained nine W-specific RAPD marker sequences (W-Bonsai, W-Mikan, W-Musashi, W-Rikishi, W-Sakura, W-Sasuke, W-Yukemuri-L, W-Yukemuri-S and BMC1-Kabuki), almost all of which contained the border regions of retrotransposons, namely portions of nested retrotransposons. We confirmed the presence of eleven out of twelve W-specific RAPD markers in the normal W chromosomes of twenty-five silkworm strains maintained in Japan. These results indicate that the W chromosomes of the strains in Japan are almost identical in type. The Zebra-W strain (T(W;3)Ze chromosome) lacked the W-Samurai and W-Mikan RAPD markers and the Black-egg-W strain (T(W;10)+(w-2) chromosome) lacked the W-Mikan RAPD marker. These results strongly indicate that the regions containing the W-Samurai and W-Mikan RAPD markers or the W-Mikan RAPD marker were deleted in the T(W;3)Ze and T(W;10)+(w-2) chromosomes, respectively, due to reciprocal translocation between the W chromosome and the autosome. This deletion apparently does not affect the expression of Fem; therefore, this deleted region of the W chromosome does not contain the putative Fem gene. PMID:16033428

  17. Enzymatic properties of alpha-amylase in the midgut and the salivary glands of mulberry moth, Glyphodes pyloalis Walker (Lepidoptera: Pyralidae).

    PubMed

    Yezdani, Elham; Sendi, Jalal Jalali; Zibaee, Arash; Ghadamyari, Mohammad

    2010-01-01

    The pyralid moth, Glyphode pyloalis Walker, is an important pest of the mulberry. Amylases are the hydrolytic enzymes that catalyze the hydrolysis of the alpha-D-(1,4)-glucan linkage in glycogen and other related carbohydrates. Laboratory-reared fifth stadium larvae were randomly selected; the midgut (MG) and the salivary glands (SG) were removed by dissection under a dissecting microscope and alpha-amylase activity was assayed using the dinitrosalicylic acid procedure. The activity of alpha-amylase in the MG and the SG were 0.011 and 0.0018 micromol/min, respectively. The optimal pH and temperature for alpha-amylase were 9 for MG at 37-40 degrees C and 10 for SG at 37 degrees C respectively. Various concentrations of compounds (NaCl, KCl, MgCl(2), Urea, EDTA, SDS and CaCl(2)) had differential effects on the enzyme activity. Plant amylase inhibitors may play an important role against insect pests. Hence, the characterization of digestive enzymes and the examination of their inhibitors may be a useful tool in future management of this important mulberry pest. PMID:20176331

  18. Identification and expression analysis of nervous wreck, which is preferentially expressed in the brain of the male silkworm moth, Bombyx mori.

    PubMed

    Kiya, Taketoshi; Iwami, M

    2011-10-01

    Sexually dimorphic neural circuits are essential for reproductive behaviour. The molecular basis of sexual dimorphism in the silkworm moth (Bombyx mori) brain, however, is unclear. We conducted cDNA subtraction screening and identified nervous wreck (Bmnwk), a synaptic growth regulatory gene, whose expression is higher in the male brain than in the female brain of the silkworm. Bmnwk was preferentially expressed in the brain at the late pupae and adult stages. In situ hybridization revealed that Bmnwk is highly expressed in the optic lobe of the male moth brain. These findings suggest that Bmnwk has a role in the development and/or maintenance of the optic lobe in the male silkworm brain. PMID:21793956

  19. 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

  20. The homeodomain transcription factors antennapedia and POU-M2 regulate the transcription of the steroidogenic enzyme gene Phantom in the silkworm.

    PubMed

    Meng, Meng; Cheng, Dao-Jun; Peng, Jian; Qian, Wen-Liang; Li, Jia-Rui; Dai, Dan-Dan; Zhang, Tian-Lei; Xia, Qing-You

    2015-10-01

    The steroid hormone ecdysone, which controls insect molting and metamorphosis, is synthesized in the prothoracic gland (PG), and several steroidogenic enzymes that are expressed specifically in the PG are involved in ecdysteroidogenesis. In this study, we identified new regulators that are involved in the transcriptional control of the silkworm steroidogenic enzyme genes. In silico analysis predicted several potential cis-regulatory elements (CREs) for the homeodomain transcription factors Antennapedia (Antp) and POU-M2 in the proximal promoters of steroidogenic enzyme genes. Antp and POU-M2 are expressed dynamically in the PG during larval development, and their overexpression in silkworm embryo-derived (BmE) cells induced the expression of steroidogenic enzyme genes. Importantly, luciferase reporter analyses, electrophoretic mobility shift assays, and chromatin immunoprecipitation assays revealed that Antp and POU-M2 promote the transcription of the silkworm steroidogenic enzyme gene Phantom (Phm) by binding directly to specific motifs within overlapping CREs in the Phm promoter. Mutations of these CREs in the Phm promoter suppressed the transcriptional activities of both Antp and POU-M2 in BmE cells and decreased the activities of mutated Phm promoters in the silkworm PG. In addition, pulldown and co-immunoprecipitation assays demonstrated that Antp can interact with POU-M2. Moreover, RNA interference-mediated down-regulation of either Antp or POU-M2 during silkworm wandering not only decreased the ecdysone titer but also led to the failure of metamorphosis. In summary, our results suggest that Antp and POU-M2 coordinate the transcription of the silkworm Phm gene directly, indicating new roles for homeodomain proteins in regulating insect ecdysteroidogenesis. PMID:26253172

  1. Novel non-autonomous transposable elements on W chromosome of the silkworm, Bombyx mori.

    PubMed

    Abe, Hiroaki; Fujii, Tsuguru; Shimada, Toru; Mita, Kazuei

    2010-01-01

    The sex chromosomes of the silkworm Bombyx mori are designated ZW (XY) for females and ZZ (XX) for males. Numerous long terminal repeat (LTR) and non-LTR retrotransposons, retroposons and DNA transposons have accumulated as strata on the W chromosome. However, there are nucleotide sequences that do not show the characteristics of typical transposable elements on the W chromosome. To analyse these uncharacterized nucleotide sequences on the W chromosome, we used whole-genome shotgun (WGS) data and assembled data that was obtained using male genome DNA. Through these analyses, we found that almost all of these uncharacterized sequences were non-autonomous transposable elements that do not fit into the conventional classification. It is notable that some of these transposable elements contained the Bombyx short interspersed element (Bm1) sequences in the elements. We designated them as secondary-Bm1 transposable elements (SBTEs). Because putative ancestral SBTE nucleotide sequences without Bm1 do not occur in theWGS data, we suggest that the Bm1 sequences of SBTEs are not carried on each element merely as a package but are components of each element. Therefore, we confirmed that SBTEs should be classified as a new group of transposable elements. PMID:20877004

  2. Two Conserved Cysteine Residues Are Required for the Masculinizing Activity of the Silkworm Masc Protein.

    PubMed

    Katsuma, Susumu; Sugano, Yudai; Kiuchi, Takashi; Shimada, Toru

    2015-10-23

    We have recently discovered that the Masculinizer (Masc) gene encodes a CCCH tandem zinc finger protein, which controls both masculinization and dosage compensation in the silkworm Bombyx mori. In this study, we attempted to identify functional regions or residues that are required for the masculinizing activity of the Masc protein. We constructed a series of plasmids that expressed the Masc derivatives and transfected them into a B. mori ovary-derived cell line, BmN-4. To assess the masculinizing activity of the Masc derivatives, we investigated the splicing patterns of B. mori doublesex (Bmdsx) and the expression levels of B. mori IGF-II mRNA-binding protein, a splicing regulator of Bmdsx, in Masc cDNA-transfected BmN-4 cells. We found that two zinc finger domains are not required for the masculinizing activity. We also identified that the C-terminal 288 amino acid residues are sufficient for the masculinizing activity of the Masc protein. Further detailed analyses revealed that two cysteine residues, Cys-301 and Cys-304, in the highly conserved region among lepidopteran Masc proteins are essential for the masculinizing activity in BmN-4 cells. Finally, we showed that Masc is a nuclear protein, but its nuclear localization is not tightly associated with the masculinizing activity. PMID:26342076

  3. Novel non-autonomous transposable elements on W chromosome of the silkworm, Bombyx mori.

    PubMed

    Abe, Hiroaki; Fujii, Tsuguru; Shimada, Toru; Mita, Kazuei

    2010-09-01

    The sex chromosomes of the silkworm Bombyx mori are designated ZW(XY) for females and ZZ (XX) for males. Numerous long terminal repeat (LTR) and non-LTR retrotransposons, retroposons and DNA transposons have accumulated as strata on the W chromosome. However, there are nucleotide sequences that do not show the characteristics of typical transposable elements on the W chromosome. To analyse these uncharacterized nucleotide sequences on the W chromosome, we used whole-genome shotgun (WGS) data and assembled data that was obtained using male genome DNA. Through these analyses,we found that almost all of these uncharacterized sequences were non-autonomous transposable elements that do not fit into the conventional classification. It is notable that some of these transposable elements contained the Bombyx short interspersed element (Bm1) sequences in the elements. We designated them as secondary-Bm1 transposable elements (SBTEs). Because putative ancestral SBTE nucleotide sequences without Bm1 do not occur in the WGS data, we suggest that the Bm1 sequences of SBTEs are not carried on each element merely as a package but are components of each element. Therefore, we confirmed that SBTEs should be classified as a new group of transposable elements. PMID:21207831

  4. Functional characterization of the vitellogenin promoter in the silkworm, Bombyx mori.

    PubMed

    Xu, J; Wang, Y Q; Li, Z Q; Ling, L; Zeng, B S; You, L; Chen, Y Z; Aslam, A F M; Huang, Y P; Tan, A J

    2014-10-01

    Genetic transformation and genome editing technologies have been successfully established in the lepidopteran insect model, the domesticated silkworm, Bombyx mori, providing great potential for functional genomics and practical applications. However, the current lack of cis-regulatory elements in B. mori gene manipulation research limits further exploitation in functional gene analysis. In the present study, we characterized a B. mori endogenous promoter, Bmvgp, which is a 798-bp DNA sequence adjacent to the 5'-end of the vitellogenin gene (Bmvg). PiggyBac-based transgenic analysis shows that Bmvgp precisely directs expression of a reporter gene, enhanced green fluorescent protein (EGFP), in a sex-, tissue- and stage-specific manner. In transgenic animals, EGFP expression can be detected in the female fat body from larval-pupal ecdysis to the following pupal and adult stage. Furthermore, in vitro and in vivo experiments revealed that EGFP expression can be activated by 20-hydroxyecdysone, which is consistent with endogenous Bmvg expression. These data indicate that Bmvgp is an effective endogenous cis-regulatory element in B. mori. PMID:24828437

  5. The silkworm Z chromosome is enriched in testis-specific genes.

    PubMed

    Arunkumar, K P; Mita, Kazuei; Nagaraju, J

    2009-06-01

    The role of sex chromosomes in sex determination has been well studied in diverse groups of organisms. However, the role of the genes on the sex chromosomes in conferring sexual dimorphism is still being experimentally evaluated. An unequal complement of sex chromosomes between two sexes makes them amenable to sex-specific evolutionary forces. Sex-linked genes preferentially expressed in one sex over the other offer a potential means of addressing the role of sex chromosomes in sexual dimorphism. We examined the testis transcriptome of the silkworm, Bombyx mori, which has a ZW chromosome constitution in the female and ZZ in the male, and show that the Z chromosome harbors a significantly higher number of genes expressed preferentially in testis compared to the autosomes. We hypothesize that sexual antagonism and absence of dosage compensation have possibly led to the accumulation of many male-specific genes on the Z chromosome. Further, our analysis of testis-specific paralogous genes suggests that the accumulation on the Z chromosome of genes advantageous to males has occurred primarily by translocation or tandem duplication. PMID:19332883

  6. Sexually biased transcripts at early embryonic stages of the silkworm depend on the sex chromosome constitution.

    PubMed

    Kawamoto, Munetaka; Koga, Hikaru; Kiuchi, Takashi; Shoji, Keisuke; Sugano, Sumio; Shimada, Toru; Suzuki, Yutaka; Katsuma, Susumu

    2015-04-10

    In the silkworm, Bombyx mori, females are heterogametic (WZ) whereas males have two Z chromosomes. Femaleness of B. mori is determined by the presence of the W chromosome, suggesting that there is a dominant feminizing gene on this chromosome. Recently, by transcriptome analysis of B. mori embryos, we discovered that a single W-chromosome-derived PIWI-interacting RNA (piRNA) is the long-sought primary determinant of femaleness in B. mori. However, sexual bias in the transcriptome of B. mori early embryos has not yet been well characterized. Using deep sequencing data from molecularly sexed RNA of B. mori embryos, we identified and characterized 157 transcripts that are statistically differentially expressed between male and female early embryos. Most of the female-biased transcripts were transposons or repeat sequences that are produced presumably from the W chromosome. Bioinformatic analysis revealed that these repetitive sequences are piRNA precursors. In contrast, male-biased genes were frequently transcribed from the Z chromosome, suggesting that dosage compensation in Z-linked genes does not occur or is incomplete at early embryonic stages. Our analysis has drawn a picture of a global landscape of sexually biased transcriptome during early B. mori embyogenesis and has suggested for the first time that most sexually biased embryonic transcripts depend on sex chromosomes. PMID:25615878

  7. 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

  8. The Expression, Purification, and Characterization of a Ras Oncogene (Bras2) in Silkworm (Bombyx mori).

    PubMed

    Lv, Zhengbing; Wang, Tao; Zhuang, Wenhua; Wang, Dan; Chen, Jian; Nie, Zuoming; Liu, Lili; Zhang, Wenping; Wang, Lisha; Wang, Deming; Wu, Xiangfu; Li, Jun; Qian, Lian; Zhang, Yaozhou

    2013-01-01

    The Ras oncogene of silkworm pupae (Bras2) may belong to the Ras superfamily. It shares 77% of its amino acid identity with teratocarcinoma oncogene 21 (TC21) related ras viral oncogene homolog-2 (R-Ras2) and possesses an identical core effector region. The mRNA of Bombyx mori Bras2 has 1412 bp. The open reading frame contains 603 bp, which encodes 200 amino acid residues. This recombinant BmBras2 protein was subsequently used as an antigen to raise a rabbit polyclonal antibody. Western blotting and real-time PCR analyses showed that BmBras2 was expressed during four developmental stages. The BmBras2 expression level was the highest in the pupae and was low in other life cycle stages. BmBras2 was expressed in all eight tested tissues, and it was highly expressed in the head, intestine, and epidermis. Subcellular localization studies indicated that BmBras2 was predominantly localized in the nuclei of Bm5 cells, although cytoplasmic staining was also observed to a lesser extent. A cell proliferation assay showed that rBmBras2 could stimulate the proliferation of hepatoma cells. The higher BmBras2 expression levels in the pupal stage, tissue expression patterns, and a cell proliferation assay indicated that BmBras2 promotes cell division and proliferation, most likely by influencing cell signal transduction. PMID:23781494

  9. 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

  10. Molecular characterization, tissue distribution, and immune reaction expression of karyopherins in the domestic silkworm (Bombyx mori).

    PubMed

    Li, J; Wang, L; Qian, C; Zhang, C F; Dai, L S; Liu, Q N; Wei, G Q; Sun, Y; Liu, D R; Zhu, B J; Liu, C L

    2015-01-01

    Karyopherins, including alpha and beta types, are transport proteins in the eukaryotic cell that carry cargoes across nuclear pore complexes into or out of the nucleus. In this study, full open reading frames of one beta and three alpha types of karyopherin were cloned from cDNA of the domestic silkworm (Bombyx mori). The one beta and three alpha types' open reading frames were 2661, 1563, 1515, and 1551 base pairs long, respectively, and coded 886, 520, 504, and 516 amino acids, respectively. The alphas all had one importin-beta-binding (IBB) domain, and eight, four, or seven armadillo/beta-catenin-like repeats. The beta had 19 HEAT repeat domains, which constructed one importin-beta-N-terminal domain and one IBB domain. The recombinant proteins were expressed in Escherichia coli cells. The molecular weight of the beta type was approximately 100 kDa, and the alphas weighed approximately 60 kDa. Phylogenic tree construction revealed that the alphas could be classified into three known karyopherin-alpha subfamilies. We detected mRNA of the four karyopherins in normal 3rd day of 5th instar larvae, and in larvae injected with Gram-positive bacteria, Gram-negative bacteria, viruses, and fungi using real-time fluorescence quantitative reverse transcriptase-polymerase chain reaction, and found that the four karyopherins were widely distributed, but their expression levels were related to tissues type, the microbe injected, and the time point. PMID:26535618

  11. Advanced technologies for genetically manipulating the silkworm Bombyx mori, a model Lepidopteran insect

    PubMed Central

    Xu, Hanfu; O'Brochta, David A.

    2015-01-01

    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

  12. Knockout silkworms reveal a dispensable role for juvenile hormones in holometabolous life cycle

    PubMed Central

    Daimon, Takaaki; Uchibori, Miwa; Nakao, Hajime; Sezutsu, Hideki; Shinoda, Tetsuro

    2015-01-01

    Insect juvenile hormones (JHs) prevent precocious metamorphosis and allow larvae to undergo multiple rounds of status quo molts. However, the roles of JHs during the embryonic and very early larval stages have not been fully understood. We generated and characterized knockout silkworms (Bombyx mori) with null mutations in JH biosynthesis or JH receptor genes using genome-editing tools. We found that embryonic growth and morphogenesis are largely independent of JHs in Bombyx and that, even in the absence of JHs or JH signaling, pupal characters are not formed in first- or second-instar larvae, and precocious metamorphosis is induced after the second instar at the earliest. We also show by mosaic analysis that a pupal specifier gene broad, which is dramatically up-regulated in the late stage of the last larval instar, is essential for pupal commitment in the epidermis. Importantly, the mRNA expression level of broad, which is thought to be repressed by JHs, remained at very low basal levels during the early larval instars of JH-deficient or JH signaling-deficient knockouts. Therefore, our study suggests that the long-accepted paradigm that JHs maintain the juvenile status throughout larval life should be revised because the larval status can be maintained by a JH-independent mechanism in very early larval instars. We propose that the lack of competence for metamorphosis during the early larval stages may result from the absence of an unidentified broad-inducing factor, i.e., a competence factor. PMID:26195792

  13. Identification and Functional Analysis of the Pre-piRNA 3' Trimmer in Silkworms.

    PubMed

    Izumi, Natsuko; Shoji, Keisuke; Sakaguchi, Yuriko; Honda, Shozo; Kirino, Yohei; Suzuki, Tsutomu; Katsuma, Susumu; Tomari, Yukihide

    2016-02-25

    PIWI-interacting RNAs (piRNAs) play a crucial role in transposon silencing in animal germ cells. In piRNA biogenesis, single-stranded piRNA intermediates are loaded into PIWI-clade proteins and cleaved by Zucchini/MitoPLD, yielding precursor piRNAs (pre-piRNAs). Pre-piRNAs that are longer than the mature piRNA length are then trimmed at their 3' ends. Although recent studies implicated the Tudor domain protein Papi/Tdrkh in pre-piRNA trimming, the identity of Trimmer and its relationship with Papi/Tdrkh remain unknown. Here, we identified PNLDC1, an uncharacterized 3'-5' exonuclease, as Trimmer in silkworms. Trimmer is enriched in the mitochondrial fraction and binds to Papi/Tdrkh. Depletion of Trimmer and Papi/Tdrkh additively inhibits trimming, causing accumulation of ∼35-40-nt pre-piRNAs that are impaired for target cleavage and prone to degradation. Our results highlight the cooperative action of Trimmer and Papi/Tdrkh in piRNA maturation. PMID:26919431

  14. Knockout silkworms reveal a dispensable role for juvenile hormones in holometabolous life cycle.

    PubMed

    Daimon, Takaaki; Uchibori, Miwa; Nakao, Hajime; Sezutsu, Hideki; Shinoda, Tetsuro

    2015-08-01

    Insect juvenile hormones (JHs) prevent precocious metamorphosis and allow larvae to undergo multiple rounds of status quo molts. However, the roles of JHs during the embryonic and very early larval stages have not been fully understood. We generated and characterized knockout silkworms (Bombyx mori) with null mutations in JH biosynthesis or JH receptor genes using genome-editing tools. We found that embryonic growth and morphogenesis are largely independent of JHs in Bombyx and that, even in the absence of JHs or JH signaling, pupal characters are not formed in first- or second-instar larvae, and precocious metamorphosis is induced after the second instar at the earliest. We also show by mosaic analysis that a pupal specifier gene broad, which is dramatically up-regulated in the late stage of the last larval instar, is essential for pupal commitment in the epidermis. Importantly, the mRNA expression level of broad, which is thought to be repressed by JHs, remained at very low basal levels during the early larval instars of JH-deficient or JH signaling-deficient knockouts. Therefore, our study suggests that the long-accepted paradigm that JHs maintain the juvenile status throughout larval life should be revised because the larval status can be maintained by a JH-independent mechanism in very early larval instars. We propose that the lack of competence for metamorphosis during the early larval stages may result from the absence of an unidentified broad-inducing factor, i.e., a competence factor. PMID:26195792

  15. Silkworm Thorn Stem Extract Targets RSK2 and Suppresses Solar UV-Induced Cyclooxygenase-2 Expression

    PubMed Central

    Kim, Jong-Eun; Lee, Ki Won

    2015-01-01

    Excessive exposure to solar UV (sUV) is associated with numerous human skin disorders, such as carcinogenesis, skin photoaging and skin inflammation. Silkworm Thorn (Cudraniatricuspidata, SW) is a plant belonging to the Moraceae family and widely present throughout Korea, China, and Japan. Most parts of the tree (including the fruit, leaf, stem, root, and bark) is consumable as a functional food or tea. In this study, we found that SW extract (SWE) inhibited the elevated expression of sUV-induced cyclooxygenase (COX)-2 levels in both HaCaT and JB6 cells. Levels of nuclear factor-κB and activator protein-1, two crucial transcription factors involved in COX-2 expression, were elevated by sUV treatment. Treatment with SWE abolished this activation. SWE also inhibited sUV-induced histone H3 phosphorylation. However, sUV-induced phosphorylation of Akt, c-Jun N-terminal kinase and p38 kinase remained unchanged in the presence of SWE. SWE inhibited RSK2 activity, and pull-down assays using SWE-Sepharose beads revealed that SWE binds directly with RSK2 in an ATP-competitive manner. These results suggest a potential for SWE to be developed as a cosmeceutical material and functional food constituent for the promotion of skin health. PMID:26506342

  16. Proteins in the Cocoon of Silkworm Inhibit the Growth of Beauveria bassiana.

    PubMed

    Guo, Xiaomeng; Dong, Zhaoming; Zhang, Yan; Li, Youshan; Liu, Huawei; Xia, Qingyou; Zhao, Ping

    2016-01-01

    Silk cocoons are composed of fiber proteins (fibroins) and adhesive glue proteins (sericins), which provide a physical barrier to protect the inside pupa. Moreover, other proteins were identified in the cocoon silk, many of which are immune related proteins. In this study, we extracted proteins from the silkworm cocoon by Tris-HCl buffer (pH7.5), and found that they had a strong inhibitory activity against fungal proteases and they had higher abundance in the outer cocoon layers than in the inner cocoon layers. Moreover, we found that extracted cocoon proteins can inhibit the germination of Beauveria bassiana spores. Consistent with the distribution of protease inhibitors, we found that proteins from the outer cocoon layers showed better inhibitory effects against B. bassiana spores than proteins from the inner layers. Liquid chromatography-tandem mass spectrometry was used to reveal the extracted components in the scaffold silk, the outermost cocoon layer. A total of 129 proteins were identified, 30 of which were annotated as protease inhibitors. Protease inhibitors accounted for 89.1% in abundance among extracted proteins. These protease inhibitors have many intramolecular disulfide bonds to maintain their stable structure, and remained active after being boiled. This study added a new understanding to the antimicrobial function of the cocoon. PMID:27032085

  17. Glucose oxidase prevents programmed cell death of the silkworm anterior silk gland through hydrogen peroxide production.

    PubMed

    Matsui, Hiroto; Kakei, Motonori; Iwami, Masafumi; Sakurai, Sho

    2011-03-01

    During pupal metamorphosis, the anterior silk glands (ASGs) of the silkworm Bombyx mori degenerate through programmed cell death (PCD), which is triggered by 20-hydroxyecdysone (20E). 20E triggers the PCD of the ASGs of day 7 fifth instar (V7) larvae but not that of V5 larvae. When V7 ASGs were cocultured with V5 ASGs in the presence of 20E, neither culture of ASGs underwent PCD. The 20E-induced PCD of V7 ASGs was also inhibited when they were incubated in conditioned medium that was prepared by incubating V5 ASGs for 48 h, an indication that V5 ASGs released an inhibitor of 20E-induced PCD during incubation. The inhibitor was purified from conditioned medium and identified as glucose oxidase (GOD). GOD catalyzes the oxidation of glucose to gluconolactone, and generates hydrogen peroxide as a byproduct. We found that hydrogen peroxide is the molecule that directly inhibits the action of 20E and may act to protect the ASGs from early execution of PCD during the feeding stage. GOD was localized in the inner cavity of the gland, and was discharged to the outside of the ASGs with the silk thread at the onset of spinning. Thus, the spinning behavior, occurring at the beginning of the prepupal period, plays an important role in controlling the time at which ASGs undergo PCD in response to 20E. PMID:21205208

  18. Mechanisms of nodule-specific melanization in the hemocoel of the silkworm, Bombyx mori.

    PubMed

    Shu, Min; Mang, Dingze; Fu, Gege Sun; Tanaka, Shiho; Endo, Haruka; Kikuta, Shingo; Sato, Ryoichi

    2016-03-01

    In the insect immune system, nodules are known to be a product of the cellular response against microorganisms and may be a preferential target for melanization. However, the mechanism of nodule-preferential melanization remains to be explored. In this study, we identified several mechanisms of nodule-preferential melanization by analyzing congregation and the activation of several factors involved in the prophenoloxidase (proPO)-activating system in the silkworm, Bombyx mori. Microorganism-binding assays revealed that B. mori larval plasma have an effective invading microorganism-surveillance network consisting of at least six pattern-recognition receptors (PRRs). We also found that a hemolymph serine proteinase, BmHP14, can bind to Saccharomyces cerevisiae. Pull-down assays showed that PRR C-type lectins form protein complexes with serine proteinase homologs, BmSPH1 and BmSPH2, which leads to the activated forms of BmSPH1 and BmSPH2 being gathered on microorganisms and trapped in nodules. Immunostaining analysis revealed that most factors in the proPO-activating system and some factors in the triggering system for antimicrobial peptide production exist in the granules of hemocytes which can gather in nodules. Western blot analysis showed that factors in the proPO-activating system are congregated in formed nodules by their concentration in plasma and aggregating hemocytes. PMID:26707571

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

    NASA Astrophysics Data System (ADS)

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

    2014-03-01

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

  20. Identification and functional characterization of the Rad23 gene of the silkworm, Bombyx mori.

    PubMed

    Xu, He-ping; Xu, Yu-song; Wang, Hua-bing; He, Da; Kawasaki, Hideki

    2010-02-01

    Rad23 is an NER (nucleotide excision repair) protein and it plays an important role in the UPP (ubiquitin-proteasome pathway). In the present study, BmRad23 (a homologous gene of Rad23 from Bombyx mori) was cloned and designated as BmRad23. The ORF (open reading frame) of the BmRad23 cDNA encoded deduced 324 amino acids with a calculated molecular mass of 36.13 kDa and an estimated pI of 4.50. The deduced amino acid sequence of the BmRad23 cDNA revealed several indispensable domains for the function of the Rad23 protein family, such as one UbL (ubiquitin-like) region domain and two UBA (ubiquitin-associated) domains. UV irradiation and treatment with chemical DNA-damaging reagent increased the expression of BmRad23. The BmRad23 gene was expressed in all the examined organs, and elevated expression was observed in testis and ovary. Northern blot and immunoblot analyses showed enhanced expression of BmRad23 after day 3 of the wandering stage in the silk gland. From the present results it is suggested that BmRad23 functions in the UPP during the silkworm metamorphosis as well as participating in the NER when the genetic material is damaged by UV irradiation and other genotoxic stresses. PMID:19203347

  1. Silkworm Thorn Stem Extract Targets RSK2 and Suppresses Solar UV-Induced Cyclooxygenase-2 Expression.

    PubMed

    Kim, Jong-Eun; Lee, Ki Won

    2015-01-01

    Excessive exposure to solar UV (sUV) is associated with numerous human skin disorders, such as carcinogenesis, skin photoaging and skin inflammation. Silkworm Thorn (Cudraniatricuspidata, SW) is a plant belonging to the Moraceae family and widely present throughout Korea, China, and Japan. Most parts of the tree (including the fruit, leaf, stem, root, and bark) is consumable as a functional food or tea. In this study, we found that SW extract (SWE) inhibited the elevated expression of sUV-induced cyclooxygenase (COX)-2 levels in both HaCaT and JB6 cells. Levels of nuclear factor-κB and activator protein-1, two crucial transcription factors involved in COX-2 expression, were elevated by sUV treatment. Treatment with SWE abolished this activation. SWE also inhibited sUV-induced histone H3 phosphorylation. However, sUV-induced phosphorylation of Akt, c-Jun N-terminal kinase and p38 kinase remained unchanged in the presence of SWE. SWE inhibited RSK2 activity, and pull-down assays using SWE-Sepharose beads revealed that SWE binds directly with RSK2 in an ATP-competitive manner. These results suggest a potential for SWE to be developed as a cosmeceutical material and functional food constituent for the promotion of skin health. PMID:26506342

  2. Proteins in the Cocoon of Silkworm Inhibit the Growth of Beauveria bassiana

    PubMed Central

    Zhang, Yan; Li, Youshan; Liu, Huawei; Xia, Qingyou; Zhao, Ping

    2016-01-01

    Silk cocoons are composed of fiber proteins (fibroins) and adhesive glue proteins (sericins), which provide a physical barrier to protect the inside pupa. Moreover, other proteins were identified in the cocoon silk, many of which are immune related proteins. In this study, we extracted proteins from the silkworm cocoon by Tris-HCl buffer (pH7.5), and found that they had a strong inhibitory activity against fungal proteases and they had higher abundance in the outer cocoon layers than in the inner cocoon layers. Moreover, we found that extracted cocoon proteins can inhibit the germination of Beauveria bassiana spores. Consistent with the distribution of protease inhibitors, we found that proteins from the outer cocoon layers showed better inhibitory effects against B. bassiana spores than proteins from the inner layers. Liquid chromatography-tandem mass spectrometry was used to reveal the extracted components in the scaffold silk, the outermost cocoon layer. A total of 129 proteins were identified, 30 of which were annotated as protease inhibitors. Protease inhibitors accounted for 89.1% in abundance among extracted proteins. These protease inhibitors have many intramolecular disulfide bonds to maintain their stable structure, and remained active after being boiled. This study added a new understanding to the antimicrobial function of the cocoon. PMID:27032085

  3. 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

  4. 'Wild Treasure' Thornless Trailing Blackberry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wild Treasure is a new trailing blackberry cultivar from the U.S. Department of Agriculture-Agricultural Research Service breeding program in Corvallis, OR, released in cooperation with Oregon State University. Wild Treasure is thornless and has high quality fruit that are very small and can be mech...

  5. Female-biased expression of odourant receptor genes in the adult antennae of the silkworm, Bombyx mori.

    PubMed

    Wanner, K W; Anderson, A R; Trowell, S C; Theilmann, D A; Robertson, H M; Newcomb, R D

    2007-02-01

    Olfaction plays an important role in the life history of insects, including key behaviours such as host selection, oviposition and mate recognition. Odour perception by insects is primarily mediated by the large diverse family of odourant receptors (Ors) that are expressed on the dendrites of olfactory neurones housed within chemosensilla. However, few Or sequences have been identified from the Lepidoptera, an insect order that includes some of the most important pest species worldwide. We have identified 41 Or gene sequences from the silkworm (Bombyx mori) genome, more than double the number of published Or sequences from the Lepidoptera. Many silkworm Ors appear to be orthologs of the 17 published tobacco budworm (Heliothis virescens) Ors indicating that many Or lineages may be conserved within the Lepidoptera. The majority of the Or genes are expressed in adult female and male antennae (determined by quantitative real-time PCR analysis), supporting their probable roles in adult olfaction. Several Or genes are expressed at high levels in both male and female antennae, suggesting they mediate the perception of common host or conspecific volatiles important to both sexes. BmOrs 45-47 group together in the same phylogenetic branch and all three are expressed at moderate female-biased ratios, six to eight times higher in female compared to male moth antennae. Interestingly, BmOrs19 and 30 appear to be expressed predominantly in female antennae, opposite to that of the published silkworm pheromone receptors BmOrs 1 and 3 that are specific to male antennae. These results suggest that BmOr19 and 30 may detect odours critical to female behaviour, such as oviposition cues or male-produced courtship pheromones. PMID:17257213

  6. 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

  7. A fluorescent single domain antibody against plumbagin expressed in silkworm larvae for fluorescence-linked immunosorbent assay (FLISA).

    PubMed

    Sakamoto, Seiichi; Pongkitwitoon, Benyakan; Sasaki-Tabata, Kaori; Putalun, Waraporn; Maenaka, Katsumi; Tanaka, Hiroyuki; Morimoto, Satoshi

    2011-05-21

    A fluorescent single-domain antibody (fluobody), a chimera of a green fluorescent protein (AcGFP) with a single chain variable fragment antibody (scFv), against plumbagin (5-hydorxy-2-methyl-1,4-naphthoquinone; PL) was successfully expressed in the hemolymph of silkworm larvae using a Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid DNA system to develop a rapid, simple, and sensitive fluorescence-linked immunosorbent assay (FLISA). In this study, two kinds of fluobody, in which the PL-scFv was fused at the N-terminus (N-fluobody) or C-terminus of AcGFP (C-fluobody), were expressed in silkworm larvae for comparative purposes. Interestingly, both fluobodies expressed in the BmNPV bacmid DNA system retained both of their original functions as an AcGFP and a PL-scFv, although the functions of the N-fluobody were found to be inferior to those of C-fluobody when they were expressed in Escherichia coli. Moreover, an improvement in the limit of quantification for PL measurement was observed in FLISA (24 ng mL(-1)) compared with conventional ELISA (0.2 µg mL(-1)). Since both the C-fluobody and N-fluobody are useful probes for FLISA and the time-, cost-consuming refolding step required in the conventional bacterial expression system can be avoided when they are expressed in the BmNPV bacmid DNA system, the silkworm expression system is useful for expressing fluobodies when developing FLISA. PMID:21442099

  8. Molecular Signatures of Reduced Nerve Toxicity by CeCl3 in Phoxim-exposed Silkworm Brains

    PubMed Central

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

    2015-01-01

    CeCl3 can reduce the damage caused by OP pesticides, in this study we used the brain of silkworms to investigate the mechanism of CeCl3 effects on pesticide resistance. The results showed that phoxim treatments led to brain damages, swelling and death of neurons, chromatin condensation, and mitochondrial damage. Normal nerve conduction was severely affected by phoxim treatments, as revealed by: increases in the contents of neurotransmitters Glu, NO, and ACh by 63.65%, 61.14%, and 98.54%, respectively; decreases in the contents of 5-HT and DA by 53.19% and 43.71%, respectively; reductions in the activities of Na+/K+-ATPase, Ca2+/Mg2+-ATPase, and AChE by 85.27%, 85.63%, and 85.63%, respectively; and increase in the activity of TNOS by 22.33%. CeCl3 pretreatment can significantly reduce such damages. Results of DGE and qRT-PCR indicated that CeCl3 treatments significantly upregulated the expression levels of CYP4G23, cyt-b5, GSTs-σ1, ace1, esterase-FE4, and β-esterase 2. Overall, phoxim treatments cause nerve tissue lesions, neuron death, and nerve conduction hindrance, but CeCl3 pretreatments can promote the expression of phoxim resistance-related genes in silkworm brains to reduce phoxim-induced damages. Our study provides a potential new method to improve the resistance of silkworms against OP pesticides. PMID:26227613

  9. Recent progress in development of transgenic silkworms overexpressing recombinant human proteins with therapeutic potential in silk glands.

    PubMed

    Itoh, Kohji; Kobayashi, Isao; Nishioka, So-Ichiro; Sezutsu, Hideki; Machii, Hiroaki; Tamura, Toshiki

    2016-01-01

    Since 2000, transgenic silkworms have been developed to produce recombinant proteins with therapeutic potential for future clinical use, including antibody preparations. Lysosomal storage diseases (LSDs) are inherited metabolic disorders caused by mutations of lysosomal enzymes associated with excessive accumulation of natural substrates and neurovisceral symptoms. Over the past few years, enzyme replacement therapy (ERT) with human lysosomal enzymes produced by genetically engineered mammalian cell lines has been used clinically to treat several patients with an LSD involving multi-organ symptoms. ERT is based on the incorporation of recombinant glycoenzymes by their binding to glycan receptors on the surface of target cells and their subsequent delivery to lysosomes. However, ERT has several disadvantages, including difficulty mass producing human enzymes, dangers of pathogen contamination, and high costs. Recently, the current authors have succeeded in producing transgenic silkworms overexpressing human lysosomal enzymes in the silk glands and the authors have purified catalytically active enzymes from the middle silk glands. Silk gland-derived human enzymes carrying high-mannose and pauci-mannose N-glycans were endocytosed by monocytes via the mannose receptor pathway and were then delivered to lysosomes. Conjugates with cell-penetrating peptides were also taken up by cultured fibroblasts derived from patients with enzyme deficiencies to restore intracellular catalytic activity and reduce the excessive accumulation of substrates in patient fibroblasts. Transgenic silkworms overexpressing human lysosomal enzymes in the silk glands could serve as future bioresources that provide safe therapeutic enzymes for the treatment of LSDs. Combining recent developments in transglycosylation technology with microbial endoglycosidases will promote the development of therapeutic glycoproteins as bio-medicines. PMID:26971553

  10. The Wilde analyst.

    PubMed

    Miller, Joel

    2013-03-01

    Oscar Wilde (1854-1900) took on the challenge of teaching us how to live artfully. From the dynamic successes and tragedies of his own life Oscar knew that everything worthy of existence is worthy of art, including its ugliness and suffering. Oscar observed much about human nature, especially his own, in an era when convention was not challenged, knowledge was taught and appearances were everything. For him, "The supreme vice is shallowness."(1) Society and psychoanalysis can still be honored and shaken by his words. The paradoxical and complex nature of Oscar's insights was as good as any coming from a thoughtful psychoanalyst. After the first two attempts to write about Oscar fell flat, it became clear that I must engage with him and try to match the unsparing commitment to explore his unconscious and interior life. In the process of creating the array of sketches of my psychoanalytic encounters with Oscar, I also found the words to describe what drew me to the field some 20 years ago-the art of psychoanalysis. PMID:23470974

  11. Leprosy in wild armadillos.

    PubMed

    Truman, Richard

    2005-09-01

    Wild nine-banded armadillos (Dasypus novemcinctus) in the south central United States are highly endemic natural hosts of Mycobacterium leprae. Surveys conducted over the last 30 years on more than 5000 animals confirm that the infection is present among armadillos in Arkansas, Louisiana, Mississippi and Texas. Highest prevalence rates are found among the animals in low-lying alluvial and coastal areas, primarily in Louisiana and Texas. Both animal density and local factors may contribute to the detectability of armadillo leprosy in those regions. Little evidence for M. leprae infection is found among armadillos elsewhere in the US range, and only a few reports relate finding the infection among animals in Central or South America. However, the issue has received only scant attention in other countries. Armadillos only recently expanded their range into the US, and leprosy was present in Texas and Louisiana prior to the arrival of armadillos. The ecological relationship between humans and armadillos with M. leprae in this region remains unclear. However, infected armadillos constitute a large reservoir of M. leprae and they may be a source of infection for some humans in this country, and perhaps in other locations across the animal's range. PMID:16248207

  12. Characterization and expression of genes involved in the ethylene biosynthesis and signal transduction during ripening of mulberry fruit.

    PubMed

    Liu, Changying; Zhao, Aichun; Zhu, Panpan; Li, Jun; Han, Leng; Wang, Xiling; Fan, Wei; Lü, Ruihua; Wang, Chuanhong; Li, Zhengang; Lu, Cheng; Yu, Maode

    2015-01-01

    Although ethylene is well known as an essential regulator of fruit development, little work has examined the role ethylene plays in the development and maturation of mulberry (Morus L.) fruit. To study the mechanism of ethylene action during fruit development in this species, we measured the ethylene production, fruit firmness, and soluble solids content (SSC) during fruit development and harvest. By comparing the results with those from other climacteric fruit, we concluded that Morus fruit are probably climacteric. Genes associated with the ethylene signal transduction pathway of Morus were characterized from M. notabilis Genome Database, including four ethylene receptor genes, a EIN2-like gene, a CTR1-like gene, four EIN3-like genes, and a RTE1-like gene. The expression patterns of these genes were analyzed in the fruit of M. atropurpurea cv. Jialing No.40. During fruit development, transcript levels of MaETR2, MaERS, MaEIN4, MaRTE, and MaCTR1 were lower at the early stages and higher after 26 days after full bloom (DAF), while MaETR1, MaEIL1, MaEIL2, and MaEIL3 remained constant. In ripening fruit, the transcripts of MaACO1 and MaACS3 increased, while MaACS1 and MaACO2 decreased after harvest. The transcripts of MaACO1, MaACO2, and MaACS3 were inhibited by ethylene, and 1-MCP (1-methylcyclopropene) upregulated MaACS3. The transcripts of the MaETR-like genes, MaRTE, and MaCTR1 were inhibited by ethylene and 1-MCP, suggesting that ethylene may accelerate the decline of MaETRs transcripts. No significant changes in the expression of MaEIN2, MaEIL1, and MaEIL3 were observed during ripening or in response to ethylene, while the expressions of MaEIL2 and MaEIL4 increased rapidly after 24 h after harvest (HAH) and were upregulated by ethylene. The present study provides insights into ethylene biosynthesis and signal transduction in Morus plants and lays a foundation for the further understanding of the mechanisms underlying Morus fruit development and ripening. PMID:25822202

  13. Characterization and Expression of Genes Involved in the Ethylene Biosynthesis and Signal Transduction during Ripening of Mulberry Fruit

    PubMed Central

    Liu, Changying; Zhao, Aichun; Zhu, Panpan; Li, Jun; Han, Leng; Wang, Xiling; Fan, Wei; Lü, Ruihua; Wang, Chuanhong; Li, Zhengang; Lu, Cheng; Yu, Maode

    2015-01-01

    Although ethylene is well known as an essential regulator of fruit development, little work has examined the role ethylene plays in the development and maturation of mulberry (Morus L.) fruit. To study the mechanism of ethylene action during fruit development in this species, we measured the ethylene production, fruit firmness, and soluble solids content (SSC) during fruit development and harvest. By comparing the results with those from other climacteric fruit, we concluded that Morus fruit are probably climacteric. Genes associated with the ethylene signal transduction pathway of Morus were characterized from M. notabilis Genome Database, including four ethylene receptor genes, a EIN2-like gene, a CTR1-like gene, four EIN3-like genes, and a RTE1-like gene. The expression patterns of these genes were analyzed in the fruit of M. atropurpurea cv. Jialing No.40. During fruit development, transcript levels of MaETR2, MaERS, MaEIN4, MaRTE, and MaCTR1 were lower at the early stages and higher after 26 days after full bloom (DAF), while MaETR1, MaEIL1, MaEIL2, and MaEIL3 remained constant. In ripening fruit, the transcripts of MaACO1 and MaACS3 increased, while MaACS1 and MaACO2 decreased after harvest. The transcripts of MaACO1, MaACO2, and MaACS3 were inhibited by ethylene, and 1-MCP (1–methylcyclopropene) upregulated MaACS3. The transcripts of the MaETR-like genes, MaRTE, and MaCTR1 were inhibited by ethylene and 1-MCP, suggesting that ethylene may accelerate the decline of MaETRs transcripts. No significant changes in the expression of MaEIN2, MaEIL1, and MaEIL3 were observed during ripening or in response to ethylene, while the expressions of MaEIL2 and MaEIL4 increased rapidly after 24 h after harvest (HAH) and were upregulated by ethylene. The present study provides insights into ethylene biosynthesis and signal transduction in Morus plants and lays a foundation for the further understanding of the mechanisms underlying Morus fruit development and ripening. PMID:25822202

  14. MicroRNA of the fifth-instar posterior silk gland of silkworm identified by Solexa sequencing

    PubMed Central

    Li, Jisheng; Ye, Lupeng; Wang, Shaohua; Che, Jiaqian; You, Zhengying; Zhong, Boxiong

    2014-01-01

    No special studies have been focused on the microRNA (miRNA) in the fifth-instar posterior silk gland of Bombyx mori. Here, using next-generation sequencing, we acquired 93.2 million processed reads from 10 small RNA libraries. In this paper, we tried to thoroughly describe how our dataset generated from deep sequencing which was recently published in BMC genomics. Results showed that our findings are largely enriched silkworm miRNA depository and may benefit us to reveal the miRNA functions in the process of silk production. PMID:26484119

  15. MicroRNA of the fifth-instar posterior silk gland of silkworm identified by Solexa sequencing.

    PubMed

    Li, Jisheng; Ye, Lupeng; Wang, Shaohua; Che, Jiaqian; You, Zhengying; Zhong, Boxiong

    2014-12-01

    No special studies have been focused on the microRNA (miRNA) in the fifth-instar posterior silk gland of Bombyx mori. Here, using next-generation sequencing, we acquired 93.2 million processed reads from 10 small RNA libraries. In this paper, we tried to thoroughly describe how our dataset generated from deep sequencing which was recently published in BMC genomics. Results showed that our findings are largely enriched silkworm miRNA depository and may benefit us to reveal the miRNA functions in the process of silk production. PMID:26484119

  16. Effects of quercetin derivatives from mulberry leaves: Improved gene expression related hepatic lipid and glucose metabolism in short-term high-fat fed mice

    PubMed Central

    Sun, Xufeng; Katsube, Takuya; Shiwaku, Kuninori

    2015-01-01

    BACKGROUND/OBJECTIVES Mulberry leaves contain quercetin derivatives, which have the effects of reducing obesity and improving lipid and glucose metabolism in mice with obesity. It is not clear whether or not mulberry leaves can directly affect metabolic disorders, in the presence of obesity, because of the interaction between obesity and metabolic disorders. The aim of the current study was to assess the direct action of quercetin derivatives on metabolic disorders in non-obese conditions in short-term high-fat diet fed mice. MATERIALS/METHODS C57BL/6N mice were fed a high-fat diet, supplemented with either 0% (control), 1%, or 3% mulberry leaf powder (Mul) or 1% catechin powder for five days. Anthropometric parameters and blood biochemistry were determined, and hepatic gene expression associated with lipid and glucose metabolism was analyzed. RESULTS Body and white fat weights did not differ among the four groups. Plasma triglycerides, total cholesterol, and free fatty acids in the 1%, 3% Mul and catechin groups did not differ significantly from those of the controls, however, plasma glucose and 8-isoprostane levels were significantly reduced. Liver gene expression of gp91phox, a main component of NADPH oxidase, was significantly down-regulated, and PPAR-α, related to β-oxidation, was significantly up-regulated. FAS and GPAT, involved in lipid metabolism, were significantly down-regulated, and Ehhadh was significantly up-regulated. Glucose-metabolism related genes, L-PK and G6Pase, were significantly down-regulated, while GK was significantly up-regulated in the two Mul groups compared to the control group. CONCLUSIONS Our results suggest that the Mul quercetin derivatives can directly improve lipid and glucose metabolism by reducing oxidative stress and enhancing β-oxidation. The 1% Mul and 1% catechin groups had similar levels of polyphenol compound intake (0.4 × 10-5 vs 0.4 × 10-5 mole/5 days) and exhibited similar effects, but neither showed dose-dependent effects on lipid and glucose metabolism or oxidative stress. PMID:25861419

  17. Mulberry-like dual-drug complicated nanocarriers assembled with apogossypolone amphiphilic starch micelles and doxorubicin hyaluronic acid nanoparticles for tumor combination and targeted therapy.

    PubMed

    Li, Ke; Liu, Hao; Gao, Wei; Chen, Mu; Zeng, Yun; Liu, Jiajun; Xu, Liang; Wu, Daocheng

    2015-01-01

    A comprehensive strategy for the preparation of mulberry-like dual-drug complicated nanocarriers (MLDC NCs) with high drug loading and adjustable dual-drug ratio was developed. First, apogossypolone (ApoG2) amphiphilic starch micelles (AASt MCs) were prepared by self-assembly process, and doxorubicin (DOX) hyaluronic acid nanoparticles (DHA NPs) were prepared by DOX absorption with excess HA by electrostatic absorption. MLDC NCs were obtained by adsorption of 8-9 DHA NPs around one AASt MC via electrostatic interaction. UV-visible and fluorescence spectrophotometers were used to measure the entrapment efficiency and loading efficiency of the two drugs. Transmission electron microscope and dynamic light scattering method were used to observe the size distribution and morphology of the particles. The tumor-targeting feature caused by HA-receptor mediation was confirmed by invitro cell uptake and invivo near-infrared fluorescence imaging. MLDC NCs were found to possess a mulberry-like shape with a dynamic size of 83.16.6nm. The final encapsulation efficiencies of ApoG2 and DOX in MLDC NCs were 941.7% and 875.8% with respect to drug-loading capacities of 13.31.2% and 13.13.7%, respectively. Almost no ApoG2 release was found within 80h and less than 30% of DOX was released into the outer phase even after 72h. Invivo fluorescence imaging revealed that MLDC NCs had highly efficient targeting and accumulation at the tumor invivo and was maintained for 96h after being injected intravenously in mice. Low LD50 for the two drugs in MLDC NCs was found after acute toxicity test. One-fifth normal dosage of the two drugs in MLDC NCs exhibited significantly higher anti-tumor efficiency in reducing tumor size compared with free drugs combination or single drug-loaded nanoparticles individually, indicating that the mulberry-like dual-drug nanoplatform has a great potential in tumor therapy. PMID:25477180

  18. Identification of two juvenile hormone inducible transcription factors from the silkworm, Bombyx mori.

    PubMed

    Matsumoto, Hitoshi; Ueno, Chihiro; Nakamura, Yuki; Kinjoh, Terunori; Ito, Yuka; Shimura, Sachiko; Noda, Hiroaki; Imanishi, Shigeo; Mita, Kazuei; Fujiwara, Haruhiko; Hiruma, Kiyoshi; Shinoda, Tetsuro; Kamimura, Manabu

    2015-09-01

    Juvenile hormone (JH) regulates many physiological processes in insects. However, the signal cascades in which JH is active have not yet been fully elucidated, particularly in comparison to another major hormone ecdysteroid. Here we identified two JH inducible transcription factors as candidate components of JH signaling pathways in the silkworm, Bombyx mori. DNA microarray analysis showed that expression of two transcription factor genes, E75 and Enhancer of split mβ (E(spl)mβ), was induced by juvenile hormone I (JH I) in NIAS-Bm-aff3 cells. Real time RT-PCR analysis confirmed that expression of four E75 isoforms (E75A, E75B, E75C and E75D) and E(spl)mβ was 3-8 times greater after JH I addition. Addition of the protein synthesis inhibitor cycloheximide did not suppress JH-induced expression of the genes, indicating that they were directly induced by JH. JH-induced expression of E75 and E(spl)mβ was also observed in four other B. mori cell lines and in larval hemocytes of final instar larvae. Notably, E75A expression was induced very strongly in larval hemocytes by topical application of the JH analog fenoxycarb; the level of induced expression was comparable to that produced by feeding larvae with 20-hydroxyecdysone. These results suggest that E75 and E(spl)mβ are general and direct target genes of JH and that the transcription factors encoded by these genes play important roles in JH signaling. PMID:25770979

  19. Retrotransposable elements on the W chromosome of the silkworm, Bombyx mori.

    PubMed

    Abe, H; Mita, K; Yasukochi, Y; Oshiki, T; Shimada, T

    2005-01-01

    The sex chromosomes of the silkworm, Bombyxmori, are designated ZW(XY) for females and ZZ(XX) for males. The W chromosome of B. mori does not recombine with the Z chromosome and autosomes and no genes for morphological characters have been mapped to the W chromosome as yet. Furthermore, femaleness is determined by the presence of a single W chromosome, regardless of the number of autosomes or Z chromosomes. To understand these interesting features of the W chromosome, it is necessary to analyze the W chromosome at the molecular biology level. Initially to isolate DNA sequences specific for the W chromosome as randomly amplified polymorphic DNA (RAPD) markers, we compared the genomic DNAs between males and females by PCR with arbitrary 10-mer primers. To the present, we have identified 12 W-specific RAPD markers, and with the exception of one RAPD marker, all of the deduced amino acid sequences of these W-specific RAPD markers show similarity to previously reported amino acid sequences of retrotransposable elements from various organisms. After constructing a genomic DNA lambda phage library of B. mori we obtained two lambda phage clones, one containing the W-Kabuki RAPD sequence and one containing the W-Samurai RAPD sequence and found that these DNA sequences comprised nested structures of many retrotransposable elements. To further analyze the W chromosome, we obtained 14 W-specific bacterial artificial chromosome (BAC) clones from three BAC libraries and subjected these clones to shotgun sequencing. The resulting assembly of sequences did not produce a single contiguous sequence due to the presence of many retrotransposable elements. Therefore, we coupled PCR with shotgun sequencing. Through these analyses, we found that many long terminal repeat (LTR) and non-LTR retrotransposons, retroposons, DNA transposons and their derivatives, have accumulated on the W chromosome as strata. These results strongly indicate that retrotransposable elements are the main structural component of the W chromosome. PMID:16093666

  20. Male-specific splicing of the silkworm Imp gene is maintained by an autoregulatory mechanism.

    PubMed

    Suzuki, Masataka G; Kobayashi, Sayaka; Aoki, Fugaku

    2014-02-01

    Sexual differentiation in the silkworm Bombyx mori is controlled by sex-specific splicing of Bmdsx, in which exons 3 and 4 are skipped in males. B. mori insulin-like growth factor II mRNA-binding protein (Imp) is a factor involved in the male-specific splicing of Bmdsx. In this study, we found that the male-specific Imp mRNA is formed as a result of the inclusion of exon 8 and the promoter-distal poly(A) site choice, whereas non-sex-specific polyadenylation occurs at the promoter-proximal poly(A) site downstream of exon 7. Recent studies revealed that Drosophila Sxl, tra in several dipteran and hymenopteran insects, and fem in Apis mellifera, play a central role in sex determination and maintain their productive mode of expression via an autoregulatory function. To determine whether Imp protein is required for the maintenance of the male-specific splicing of its own pre-mRNA, we knocked down endogenous Imp in male cells and assessed the male-specific splicing of an exogenous Imp minigene. Knockdown of endogenous Imp inhibited the male-specific splicing of the Imp minigene transcript. In contrast, overexpression of Imp in female cells induced the male-specific splicing of the Imp minigene transcript. Moreover, deletion of adenine-rich (A-rich) sequences located downstream of the proximal poly(A) site repressed the male-specific splicing of the Imp minigene transcript. Finally, gel shift analysis demonstrated that Imp binds to the A-rich sequences. These data suggest that Imp binds to the A-rich sequences in its own pre-mRNA to induce the male-specific splicing of its pre-mRNA. PMID:24231282