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posted Mar 4, 2011, 8:23 AM by rajesh gk

Title

Characterization of recombinant Autographa californica nuclear polyhedrosis virus (NPV) expressing the beta-galactosidase gene in both Sf21 and Bm5 cells by Bombyx mori NPV p143 helicase gene.

Authors

Jin BR, Yoon HJ, Choudary PV, Kang SK.

Laboratory of Insect Genetic Engineering, National Sericulture & Entomology Research Institute, Suwon, Korea.

Journal

Mol Cells. 1997 Dec 31;7(6):762-8.

Abstract

Genomic DNA of recombinant AcNPV expressing beta-galactosidase was cotransfected with p143 helicase gene of BmNPV into Sf21 cells. Ac-Bm hybrid viruses capable of replicating in both Bm5 and Sf21 cells were isolated. Ac-Bm hybrid viruses expressing beta-galactosidase either at the highest (Ac-Bm hybrid virus-HE) or lowest (Ac-Bm hybrid virus-LE) level were chosen for the characterization of beta-galactosidase expression in Bm5 and Sf21 cells. Expression level of beta-galactosidase and replication of Ac-Bm hybrid virus-HE in Sf21 cells were nearly identical to those of recombinant AcNPV. Furthermore, replication of Ac-Bm hybrid virus-HE in Bm5 cells was similar to that of wild-type BmNPV, and Ac-Bm hybrid virus-HE clearly expressed beta-galactosidase in Bm5 cells. However, expression of beta-galactosidase by Ac-Bm hybrid virus-HE in Bm5 cells was significantly lower than that expressed in Sf21 cells. The titer of Ac-Bm hybrid virus-HE determined by plaque assays in Bm5 cells was similar to that determined in Sf21 cells, but the plaque size formed by Ac-Bm hybrid virus-HE in Bm5 cells was apparently smaller than that formed in Sf21 cells. In addition, expression levels and virus titers of Ac-Bm hybrid virus-LE in Sf21 and Bm5 were significantly lower than those of Ac-Bm hybrid virus-HE. Therefore, DNA sequences were determined for the region of the p143 gene controlling the host range in Ac-Bm hybrid viruses. The results showed that the deduced amino acid sequences of Ac-Bm hybrid virus-HE were almost identical to those of BmNPV. There were differences only in amino acids at positions 461 and 470, whereas those of Ac-Bm hybrid virus-LE were different at position 461, 470, 514, and 528 from those of BmNPV. In conclusion, our results clearly demonstrated that Ac-Bm hybrid virus-HE has an additional advantage of expanded host range for producing recombinant proteins.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/9509418 PMID: 9509418 [PubMed - indexed for MEDLINE]

 

Title

Nucleotide sequence and transcriptional analysis of the DNA polymerase gene of Bombyx mori nuclear polyhedrosis virus.

Authors

Chaeychomsri S, Ikeda M, Kobayashi M.

Laboratory of Sericulture and Entomoresources, School of Agriculture, Nagoya University, Japan.

Journal

Virology. 1995 Jan 10;206(1):435-47.

Abstract

A gene encoding a putative DNA polymerase (pol) of Bombyx mori nuclear polyhedrosis virus (BmSNPV) was cloned and sequenced. The gene included an open reading frame (ORF) encoding a polypeptide of 988 amino acids with a predicted molecular mass of 114.65 kDa. The deduced amino acid sequence of the BmSNPV pol ORF showed an overall identity of 96 and 45% to those of the Autographa californica NPV (AcMNPV) pol ORF and the Lymantria dispar NPV pol ORF, respectively, and contained sequences conserved in a variety of eukaryotic and viral replicative DNA polymerases. The BmSNPV pol lacked a canonical TATAA element but contained a G+C-rich sequence in the transcriptional initiation region. Analyses by Northern blot hybridization, RNase protection assay, primer extension, and 3' and 5' RACE (rapid amplification of cDNA ends) showed that at least seven different transcripts of approximately 3.1 kb that shared a common 3' end were expressed from the BmSNPV pol. The expression of these transcripts from BmSNPV pol was regulated differentially during virus infection. Transcription of five of the seven species initiated in the close vicinity of and within the motif 5'-GCGTGCT-3'. One transcript placed its initiation site within the motif 5'-AGAGCGT-3' and the remaining one within the motif 5'-GGCGGTGG-3'. The motifs 5'-GCGTGCT-3' and 5'-AGAGCGT-3' have been identified in pol and other genes of AcMNPV as conserved sequences containing transcriptional initiation sites, whereas the motif 5'-GGCGGTGG-3', which is arranged as a direct repeat in BmSNPV pol but not in AcMNPV pol, has not been defined as the sequence responsible for transcriptional initiation sites. The BmSNPV pol transcripts were detectable at 2 hr postinfection (p.i.), peaked at 10 hr p.i., and declined to a low level by 18 hr p.i. The expression of BmSNPV pol was not inhibited but delayed dramatically by the protein synthesis inhibitor cycloheximide upon treatment of infected cells, whereas aphidicolin, an inhibitor of DNA polymerase, inhibited BmSNPV pol transcription. These results suggest a complicated and unique mechanism for the regulation of BmSNPV pol expression.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/7831799 PMID: 7831799 [PubMed - indexed for MEDLINE]

 

Title

Dominant lethals induced by Dithane M-45 in silkworm Bombyx mori.

Authors

Vasudev V, Subramanya G, Krishnamurthy NB.

Department of Studies in Zoology, University of Mysore, India.

Journal

Environ Res. 1994 Apr;65(1):145-8.

Abstract

Dithane M-45, a ethylenebis(dithiocarbamate) fungicide widely used in agriculture, including moriculture and sericulture, was tested for its efficacy in inducing dominant lethals in Bombyx mori. A polyvotine race of Pure Mysore was used for the studies and the topical application method was employed. After treatment with sublethal concentrations of 5, 10, and 20 g/50 larvae, the results have revealed that all of the concentrations could induce significant dominant lethals in a dose-dependent manner compared to controls. The results are discussed in the light of precautionary measures in the use of Dithane M-45 in the sericulture industry.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/8162881 PMID: 8162881 [PubMed - indexed for MEDLINE]

 

Title

Susceptibility of three Eco-races of tropical Tasar silkworm to Antheraea mylitta cytoplasmic polyhedrosis virus (AmCPV)

Authors

Singh1, G.P.*, Srivastava, A.K., Prakash, B., Ojha, N.G., Suryanarayana, N., Zeya, S.B.,

Journal

Caspian J. Env. Sci., 6, 161-165. ( 2008 )

Abstract

Pathogenic infection in tasar silkworm, Antheraea mylitta Drury. is common and there is a breed specific response regarding tolerance. Observations reveal the possibility of differential response by haemocytes to microbial infection in different breeds. Hence, the susceptibility of three eco-races of tasar silkworm viz. Daba, Sarihan and Raily to Antheraea mylitta Cytoplasmic Polyhedrosis Virus (AmCPV) infection and difference in total haemocyte counts were tested. The survival of Daba, Sarihan and Raily eco-races was significantly different (p < 0.05) when challenged with the same concentration (1 x 105 polyhedra/ml) of AmCPV. Daba eco-race was more tolerant to the AmCPV infection having higher survival (66.7%) and LC50 values (1000893.1796 polyhedra/ml) of AmCPV followed by Sarihan eco-race (50.7% survival and LC50 value of AmCPV 187203.6168 polyhedra/ml.) and Raily eco-race (25.3% survival and LC50 value of AmCPV 5176.37 polyhedra/ml.). Difference in total haemocyte count i.e. higher in tolerant (Daba) and lower in susceptible (Raily) eco-race may be in response to difference to their susceptibility to AmCPV infection.

Citation

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Title

Purification and characterization of antiviral protein from silkworm fecal matter

Authors

Neelagund, S.E.*, Ingalhalli, S.S., Savanurmath, C.J., Hinchigeri3, S.B., Hiremath, M. B

Journal

Caspian J. Env. Sci., 5, 77-85. , ( 2007 ),

Abstract

Antiviral proteins (AVP), present in silkworm fecal matter, show activity against nuclear polyhedrosis virus (NPV) in vitro and in vivo. The extract of silkworm fecal matter prepared in phosphate buffer solution of pH 7.5 was subjected to 50% solid ammonium sulfate precipitation to enrich AVP, then which was dialyzed. The dialysate was applied to the column containing silica gel-G, the column elutes were purified by gelfiltration chromatography. The gelfiltration pattern gave three protein peaks A, B and C. The protein obtained from peak fractions of peak A is found to be active against NPV in vitro. Whereas the proteins were obtained from peak fractions of peaks C and B were not shown activity against NPV in vitro. The peak A fractions were collected and further purified by High Pressure Liquid Chromatography (HPLC) using C4 column. Purified AVP of SDS-polyacrylamide gel electrophoresis (SDS-PAGE) resulted in two protein bands with the molecular mass of 23 KD and 16 KD. Thymol sulphuric acid method of carbohydrate staining indicated that both of these protein bands are glycoproteins. AVP activity is determined in vitro by precipitation reaction. In vivo activity of the AVP is confirmed by conducting the bioassay in silkworms.

Citation

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Title

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

[Article in Chinese]

Authors

Cao Y, Xiao QX, Huang YD, Ge CB, Huang ZR, Liu LS.

Department of Sericulture and Fashion Design, South China Agricultural University, Guanzhou 510642, China.

Journal

Yi Chuan Xue Bao. 2000;27(11):972-81.

Abstract

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

Citation

http://www.ncbi.nlm.nih.gov/pubmed/11209691 PMID: 11209691 [PubMed - indexed for MEDLINE]

 

Title

[Study on the biology feature and pathogenicities to silkworm of a microsporidium isolated from Barathra brassicae L.]

[Article in Chinese]

Authors

Zheng X, Zhou Y, Huang B, Lu K, Huang X.

Institute of Sericulture, Guangdong Academy of Agricultural Sciences, Guangzhou 510640.

Journal

Wei Sheng Wu Xue Bao. 2000 Oct;40(5):540-4.

Abstract

A microsporidium (called as Bab-M) was isolated from Barathra brassicae L. captured from suburban vegetable plot of Guang Zhou. The spores were long-ovoid in shape and 4.02 +/- 0.36 microns x 1.99 +/- 0.36 microns in size. Immunologically the microsporidium shared spore surface specific antigen(s) with N. bombycis. The ultrastructure and life cycle of Bab-M were similar to that of N. b.. The rate of transovarian transmission was high. The initial conclusion was that Bab-M should be referred to as Nosema bombycis, but there was variation between them.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/12548768 PMID: 12548768 [PubMed - indexed for MEDLINE]

 

Title

Immune stimulation in the silkworm, Bombyx mori L., by CpG oligodeoxynucleotides.

Authors

Kim I, Kim SH, Lee YS, Yun EK, Lee HS, Kim JW, Ryu KS, Kang PD, Lee IH.

Department of Sericulture & Entomology, National Institute of Agricultural Science and Technology, Rural Development Administration, Suwon, South Korea.

Journal

Arch Insect Biochem Physiol. 2004 Jan;55(1):43-8.

Abstract

Synthetic ODNs containing unmethylated CpG dinucleotides are known to stimulate immune responses in vertebrates, but so far the effect has not been studied in insects. In this report, we describe an induction of immune response following injection of oligodeoxynucleotides (ODNs) into the insect hemocoel. The fifth instar silkworm (Bombyx mori L.) larvae were injected with several synthetic ODNs containing variable number of unmethylated CpG motifs, heat-denatured genomic DNA of B. mori itself, or intact genomic DNA to observe a new induction pattern in the insect immune mechanism. When the induction of immune response was examined based on the expression rates of genes for antibacterial peptides such as attacin and cecropin, we could confirm that it was triggered upon injection of ODNs. The expression was, however, neither dependent on numbers of CpG motifs nor methylation of CpGs in ODNs. Furthermore, it was confirmed that the presence of CpG in ODN was not involved in the induction pattern of insect immunity caused by ODNs, although it has been reported that vertebrates respond in a specific manner against invading ODNs containing CpG dinucleotides. In addition, insect immunity was not stimulated by injection of intact DNA from host. In contrast, the injection of denatured genomic DNA provoked the host immune reaction. Taken together, our data suggest that foreignness of ODNs or DNA might be a key factor in the induction of insect immunity. Copyright 2003 Wiley-Liss, Inc.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/14691962 PMID: 14691962 [PubMed - indexed for MEDLINE]

 

Title

Characterization of Antheraea pernyi nucleopolyhedrovirus p11 gene, a homologue of Autographa californica nucleopolyhedrovirus orf108.

Authors

Shi SL, Pan MH, Lu C.

Key Sericultural Laboratory of Agriculture Ministry, College of Sericulture and Biotechnology, Southwest University, Chongqing, 400716, China.

Journal

Virus Genes. 2007 Aug;35(1):97-101. Epub 2006 Oct 27.

Abstract

Antheraea pernyi nucleopolyhedrovirus (ApNPV) p11 gene is 309 bp long, potentially encoding 102 amino acids with a predicted molecular weight of 11.2 kDa. ApNPV p11 gene was cloned into the prokaryotic expression vector pQE-30 and P11 was expressed in E. coli M15. Polyclonal antiserum was made against 6xHis tagged P11 protein expressed in E. coli M15. P11 gene transcription was detected as early as 36 h post-infection (p.i.) in tussah pupa and remain at high level up to 96 h p.i. Structural localization revealed that P11 protein was present in polyhedral inclusion bodies (PIB) dilute alkaline saline (DAS) pellet (P) fractions and occlusion-derived virus (ODV), but not in PIB DAS supernatant (S) fractions and budded virus (BV). These results indicated that P11 was associated with ApNPV structure.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/17072759 PMID: 17072759 [PubMed - indexed for MEDLINE]

 

Title

Effects of Wolbachia in the uzifly, Exorista sorbillans, a parasitoid of the silkworm, Bombyx mori. Free PMC Article

Authors

Puttaraju HP, Prakash BM.

Seribiotechnology Research Laboratory, Department of Sericulture, Bangalore University, Bangalore-560 056, India. puttarajuhp@hotmail.com

Journal

J Insect Sci. 2005 Nov 11;5:30.

Abstract

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.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/17119612 PMID: 17119612 [PubMed - indexed for MEDLINE]PMCID: PMC1615237

 

Title

[Identification of white muscardin silkworms by infrared spectroscopy]

[Article in Chinese]

Authors

Ji XL, Gai YP, Mu ZM, Wang GX, Ji SL.

Department of Sericulture, Shandong Agricultural University, Taian 271018, China.

Journal

Guang Pu Xue Yu Guang Pu Fen Xi. 2007 Jan;27(1):66-9.

Abstract

The infrared spectra of the ethanol extracts of well-living silkworms and white muscardin silkworms of different seasons and breeds were analyzed by means of the sequential analysis in which two indexes, i. e. common peak ratio and variant peak ratio, were applied. The results showed that the ethanol extracts of white muscardin silkworm have a stable and distinct infrared spectrum. The spectral differences of the ethanol extracts between white muscardin silkworms and well-living silkworms were so obvious that the common peak ratio of them was no more than 63. 0%, and the variant peak ratio amounted to 41. 2%. The spectra of different breeds and seasons conformed with each other with a few small differences. The minimum common peak ratio of the spectra of different breeds was 76. 0%, and the maximal ratio was 92. 0%. The common peak ratio of the spectra of different seasons was 73. 1%. Infrared spectrometry was proved to be good for the identification of white muscardin silkworms and the differentiation of white muscardin silkworms of different breeds and seasons.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/17390651 PMID: 17390651 [PubMed - indexed for MEDLINE

 

Title

A proteomic-based approach for the characterization of some major structural proteins involved in host-parasite relationships from the silkworm parasite Nosema bombycis (Microsporidia).

Authors

Wang JY, Chambon C, Lu CD, Huang KW, Vivarès CP, Texier C.

Equipe Parasitologie Moléculaire et Cellulaire, LBP, UMR CNRS 6023, Université Blaise Pascal, Aubière, France.

Journal

Proteomics. 2007 May;7(9):1461-72.

Abstract

Nosema bombycis is the causative agent of the silkworm Bombyx mori pebrine disease which inflicts severe worldwide economical losses in sericulture. Little is known about host-parasite interactions at the molecular level for this spore-forming obligate intracellular parasite which belongs to the fungi-related Microsporidia phylum. Major microsporidian structural proteins from the spore wall (SW) and the polar tube (PT) are known to be involved in host invasion. We developed a proteomic-based approach to identify few N. bombycis proteins belonging to these cell structures. Protein extraction protocols were optimized and four N. bombycis spore protein extracts were compared by SDS-PAGE and 2-DE to establish complementary proteomic profiles. Three proteins were shown to be located at the parasite SW. Moreover, 17 polyclonal antibodies were raised against major N. bombycis proteins from all extracts, and three spots were shown to correspond to polar tube proteins (PTPs) by immunofluorescent assay and transmission electron microscopy immunocytochemistry on cryosections. Specific patterns for each PTP were obtained by MALDI-TOF-MS and MS/MS. Peptide sequence tags were deduced by de novo sequencing using Peaks Online and DeNovoX, then evaluated by MASCOT and SEQUEST searches. Identification parameters were higher than false-positive hits, strengthening our strategy that could be enlarged to a nongenomic context.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/17407187 PMID: 17407187 [PubMed - indexed for MEDLINE]

 

Title

Frequency of infection with A and B supergroup Wolbachia in insects and pests associated with mulberry and silkworm. Free Article

Authors

Prakash BM, Puttaraju HP.

Laboratory of Seribiotechnology, Department of Sericulture and Life Sciences, Bangalore University, Bangalore 560 056, India.

Journal

J Biosci. 2007 Jun;32(4):671-6.

Abstract

Wolbachia is a ubiquitous, Gram-negative,vertically transmitted, alpha-proteobacterium that causes an array of reproductive abnormalities including cytoplasmic incompatibility, feminization of genetic males, parthenogenesis in a number of insect species, among others. Wolbachia is now being exploited as an agent for pest and vector control. Previous surveys indicated that it is commonly seen in 16-76% of arthropods. In this paper, using polymerase chain reaction assay based on specific amplification of the ftsZ -A and -B supergroup Wolbachia gene fragments, we found that 30% of insects and pests screened were positive for Wolbachia. Among them 66.7% harbour double Wolbachia infection, while 33.3 % harbour single Wolbachia infection. These results indicate widespread infection with both double and single Wolbachia, and provide a wealth of information to exploit this endobacterium for the management of pests and vectors.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/17762140 PMID: 17762140 [PubMed - indexed for MEDLINE]

 

Title

[The analysis of bro genes of GD isolate of Bombyx mori nucleopolyhedrovirus]

[Article in Chinese]

Authors

Pang M, Pan GQ, Li T, Wang X, Zhou ZY.

The Key Laboratory of Sericulture of Agricultural Ministry, Southwest University, Chongqing 400715, China.

Journal

Bing Du Xue Bao. 2007 Nov;23(6):485-9.

Abstract

BmNPV GD isolate from China was plaque-purified and four bro genes were cloned termed as bro-a,b,c,d. The obtained sequences were aligned to the related sequences in GenBank and the BmNPV CQ1 isolate preserved in our laboratory. Compared with genome sequences of BmNPV T3 isolate, bro genes of GD isolate housed insertion and deletion, and the changes of amino acid mainly occured at the N terminal of corresponding protein. The phylogenetic analysis of bro genes indicated that GD bro-d gene belongs to subgroup A together with T3, CQ1 bro-d and SC7 bro- III; GD bro-a, c genes belong to subgroup B together with T3, CQ1 bro-a, c and SC7 bro-II; GD bro-b gene belongs to subgroup C together with T3, CQ1 bro-b, e and SC7 bro-I. The evolutionary relationship of bro genes showed vague relevance to their geographical location. The distribution character of bro genes in four BmNPV isolates is coincidence with the KANG's theory that the bro-d plays an irreplaceable functional role(s) during viral replication, while bro-a and bro-c functionally complement each other. Meanwhile, we postulate that 3 bro genes in SC7 isolate is probable the most simple form of bro genes.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/18092688 PMID: 18092688 [PubMed - indexed for MEDLINE]

 

Title

Identification of a novel spore wall protein (SWP26) from microsporidia Nosema bombycis.

Authors

Li Y, Wu Z, Pan G, He W, Zhang R, Hu J, Zhou Z.

The Key Sericultural Laboratory of Agricultural Ministry, Southwest University, Beibei, Chongqing 400716, PR China.

Journal

Int J Parasitol. 2009 Mar;39(4):391-8. Epub 2008 Sep 27.

Abstract

Microsporidia are obligate intracellular parasites related to fungi with resistant spores against various environmental stresses. The rigid spore walls of these organisms are composed of two major layers, which are the exospore and the endospore. Two spore wall proteins (the endosporal protein-SWP30 and the exosporal protein-SWP32) have been previously identified in Nosema bombycis. In this study, using the MALDI-TOF-MS technique, we have characterised a new 25.7-kDa spore wall protein (SWP26) recognised by monoclonal antibody 2G10. SWP26 is predicted to have a signal peptide, four potential N-glycosylation sites, and a C-terminal heparin-binding motif (HBM) which is known to interact with extracellular glycosaminoglycans. By using a host cell binding assay, recombinant SWP26 protein (rSWP26) can inhibit spore adherence by 10%, resulting in decreased host cell infection. In contrast, the mutant rSWP26 (rDeltaSWP26, without HBM) was not effective in inhibiting spore adherence. Immuno-electron microscopy revealed that this protein was expressed largely in endospore and plasma membrane during endospore development, but sparsely distributed in the exospore of mature spores. The present results suggest that SWP26 is a microsporidia cell wall protein that is involved in endospore formation, host cell adherence and infection in vitro. Moreover, SWP26 could be used as a good prospective target for diagnostic research and drug design in controlling the silkworm, Bombyx mori, pebrine disease in sericulture.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/18854188 PMID: 18854188 [PubMed - indexed for MEDLINE]

 

Title

Protein profile of Nomuraea rileyi spore isolated from infected silkworm.

Authors

Qin L, Liu X, Li J, Chen H, Yao Q, Yang Z, Wang L, Chen K.

Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang 212013, Jiangsu Province, People's Republic of China. qinlvgao@163.com

Journal

Curr Microbiol. 2009 Jun;58(6):578-85. Epub 2009 Mar 14.

Abstract

Nomuraea rileyi (N. rileyi) is the causative agent of the silkworm, Bombyx mori, green muscardine which can cause severe worldwide economical loss in sericulture. Little is known about N. rileyi at the protein level for this entomopathogenic parasite which belongs to the Ascomycota. Here, we employed proteomic-based approach to identify proteins of N. rileyi spores collected from the dead silkworm. In all, 252 proteins were separated by two-dimensional gel electrophoresis (2-DE), and were subjected to mass spectrometry (MS) analysis, 121 proteins have good MS signal, and 24 of them were identified due to unavailability of genomic information from N. rileyi. This data will be helpful in understanding the biochemistry of N. rileyi.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/19288155 PMID: 19288155 [PubMed - indexed for MEDLINE]

 

Title

Molecular analysis of divergence in tachinid Uzi (Exorista sorbillans) populations in India.

Authors

Chatterjee SN, Taraphdar T, Mohandas TP.

SeriBiotech Laboratory, Central Silk Board, Kodathi Campus, Sarjapur Road, Carmelram, 560035 Bangalore, Karnataka, India. shankerc99@yahoo.com

Journal

Genetica. 2005 Sep;125(1):1-15.

Abstract

Exorista sorbillans is a tachinid endoparasitoid of silkworm, Bombyx mori, and is globally known as uzi. It causes economic injury to the cocoon crop in silkworm cultivating areas of India, except those above 400 m above mean sea level (AMSL) in the foothills of the Himalayas (Darjeeling). It is reported that the sericulture tract of south India became infected with this pest only since 1980 through an accidental transportation of cocoons from West Bengal. To ascertain whether the genome of this parasitoid is differentiating into discrete gene pools in contrasting geo-climatic conditions, molecular profiling of four populations (Es (Annatapur), Es(Ramanagaram), Es (Channapatna) and Es(Kodathi) from south India and Es(Murshidabad) from Murshidabad, West Bengal was undertaken with 13 ISSR, 3 RAPD and six non-random primers designed from various repeat sequences of B. mori . MANOVA indicated significance for the Roy's largest root estimate (55.4; F =18.47; p = 0.002) for the variability contributed by the replication. Further, hierarchical clustering done on the basis of Euclidean distance matrix and Nei's unbiased Phylip clustering put Es(Murshidabad) at the maximum distance from those of south India and 29 markers could also be identified which significantly differentiateEs(Murshidabad) from others. However, Nei's statistics for gene diversity in sub-populations reveal considerably high gene-flow (3.44 and 2.51) among the populations around Bangalore. The gene-flow between Es(Murshidabad) and other population is lowest but cannot be ignored. The comparison of endosymbiont specific 16SrRNA and fts Z gene (partial) sequences through clustalW (gcgMSF) revealed a closer relationship of Es(Murshidabad) with Es(Annatapur) and Es (Ramanagaram) and is not congruent with the relationships discussed above. The significance of this maiden study with a tachinid fly-pest is discussed in the context of understanding the diversification of Uzi fly-pest and also establishing this pest as a relevant biological material for studying microevolution in future.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/16175450 PMID: 16175450 [PubMed - indexed for MEDLINE]

 

Title

Gene organization and complete sequence of the Hyphantria cunea nucleopolyhedrovirus genome. Free Article

Authors

Ikeda M, Shikata M, Shirata N, Chaeychomsri S, Kobayashi M.

 

Laboratory of Sericulture and Entomoresources, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan.

Journal

J Gen Virol. 2006 Sep;87(Pt 9):2549-62.

Abstract

The whole-genome sequence of the Hyphantria cunea nucleopolyhedrovirus (HycuNPV) was analysed. The entire nucleotide sequence of the HycuNPV genome was 132 959 bp long, with a G+C content of 45.1 mol%. A total of 148 open reading frames (ORFs) consisting of more than 50 aa were encoded by the genome. HycuNPV shares more than 122 ORFs with other lepidopteran group I NPVs, including Autographa californica MNPV, Bombyx mori NPV, Choristoneura fumiferana MNPV (CfMNPV), Choristoneura fumiferana defective NPV, Epiphyas postvittana MNPV and Orgyia pseudotsugata MNPV (OpMNPV). Six ORFs are identified as being unique to HycuNPV. Most of the HycuNPV ORFs showed higher similarity to CfMNPV and OpMNPV ORFs than to those of the other group I NPVs. HycuNPV encodes two conotoxin-like homologues (ctls), which are observed only in OpMNPV in group I NPVs. HycuNPV encodes three inhibitors of apoptosis (iaps), hycu-iap-1, hycu-iap-2 and hycu-iap-3, a feature that it shares only with CfMNPV. In addition, six homologous regions (hrs) are identified in the HycuNPV genome. These hrs are located in regions similar to those of the OpMNPV hrs, but different from most of the CfMNPV hrs. Based on the close phylogenetic relationship and conservation of group I NPV-specific genes, such as gp64, ie-2 and ptp-1, it is concluded that HycuNPV belongs to the group I NPVs and is most similar to CfMNPV or OpMNPV.

PMID: 16894193 [PubMed - indexed for MEDLINE]

Citation

http://www.ncbi.nlm.nih.gov/pubmed/16894193

 

Title

Inactivation and mechanisms of chlorine dioxide on Nosema bombycis.

Authors

Wang Z, Liao F, Lin J, Li W, Zhong Y, Tan P, Huang Z.

 

Department of Sericulture Science, South China Agricultural University, Guangzhou 510642, China.

Journal

J Invertebr Pathol. 2010 Jun;104(2):134-9. Epub 2009 Dec 29.

Abstract

Biological tests demonstrated that the inactivation of Nosema bombycis (N. bombycis) spores by chlorine dioxide (ClO(2)) occurs very fast and is highly sensitive. The lowest effective inactivation dosage and time was 15mg/mL for 30min. The inactivation of spores was additionally verified by using double color fluorescence stain and spore germination testing. A series of biological changes, including a large number of substrates that were leaked out from the spores included proteins, DNA, polysaccharide, K(+), and Ca(2+), occurred a short time after N. bombycis spores were treated with ClO(2). In addition, the lipid of spores was disrupted and ATPase activity was inhibited, which resulted in the destruction of the inner structure of the spores.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/20036671 PMID: 20036671 [PubMed - in process]

 

Title

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

Authors

Huang L, Cheng T, Xu P, Cheng D, Fang T, Xia Q.

 

Institute of Sericulture and Systems Biology, Southwest University, Chongqing, China.

Journal

PLoS One. 2009 Dec 1;4(12):e8098.

Abstract

Host-pathogen interactions are complex relationships, and a central challenge is to reveal the interactions between pathogens and their hosts. Bacillus bombysepticus (Bb) which can produces spores and parasporal crystals was firstly separated from the corpses of the infected silkworms (Bombyx mori). Bb naturally infects the silkworm can cause an acute fuliginosa septicaemia and kill the silkworm larvae generally within one day in the hot and humid season. Bb pathogen of the silkworm can be used for investigating the host responses after the infection. Gene expression profiling during four time-points of silkworm whole larvae after Bb infection was performed to gain insight into the mechanism of Bb-associated host whole body effect. Genome-wide survey of the host genes demonstrated many genes and pathways modulated after the infection. GO analysis of the induced genes indicated that their functions could be divided into 14 categories. KEGG pathway analysis identified that six types of basal metabolic pathway were regulated, including genetic information processing and transcription, carbohydrate metabolism, amino acid and nitrogen metabolism, nucleotide metabolism, metabolism of cofactors and vitamins, and xenobiotic biodegradation and metabolism. Similar to Bacillus thuringiensis (Bt), Bb can also induce a silkworm poisoning-related response. In this process, genes encoding midgut peritrophic membrane proteins, aminopeptidase N receptors and sodium/calcium exchange protein showed modulation. For the first time, we found that Bb induced a lot of genes involved in juvenile hormone synthesis and metabolism pathway upregulated. Bb also triggered the host immune responses, including cellular immune response and serine protease cascade melanization response. Real time PCR analysis showed that Bb can induce the silkworm systemic immune response, mainly by the Toll pathway. Anti-microorganism peptides (AMPs), including of Attacin, Lebocin, Enbocin, Gloverin and Moricin families, were upregulated at 24 hours post the infection.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/19956592

PMID: 19956592 [PubMed - indexed for MEDLINE]PMCID: PMC2780328

 

Title

Natural selection maintains the transcribed LTR retrotransposons in Nosema bombycis.

Authors

Xiang H, Pan G, Zhang R, Xu J, Li T, Li W, Zhou Z, Xiang Z.

 

Institute of Sericulture and Systems Biology, Southwest University, Beibei, Chongqing 400715, China.

Journal

J Genet Genomics. 2010 May;37(5):305-14.

Abstract

Eight intact LTR retrotransposons (Nbr1-Nbr8) have been previously characterized from the genome of Nosema bombycis, a eukaryotic parasite with a compact and reduced genome. Here we describe six novel transcribed Nbr elements (Nbr9-Nbr14) identified through either cDNA library or RT-PCR. Like previously determined ones, all of them belong to the Ty3/Gypsy superfamily. Retrotransposon diversity and incomplete domains with insertions (Nbr12), deletions (Nbr11) and in-frame stop codons in coding regions (Nbr9) were detected, suggesting that both defective and loss events of LTR retrotransposon have happened in N. bombycis genome. Analysis of selection showed that strong purifying selection acts on all elements except Nbr11. This implies that selective pressure keeps both these Nbrs and their functions in genome. Interestingly, Nbr11 is under positive selection and some positively selected codons were identified, indicating that new functionality might have evolved in the Nbr11 retrotransposon. Unlike other transposable elements, Nbr11 has integrated into a conserved syntenic block and probably resulted in the inversion of both flanking regions. This demonstrates that transposable element is an important factor for the reshuffling and evolution of their host genomes, and may be maintained under natural selection. (c) 2010 Institute of Genetics and Developmental Biology and the Genetics Society of China. Published by Elsevier Ltd. All rights reserved.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/20513631 PMID: 20513631 [PubMed - in process]

 

Title

Open reading frame 60 of the Bombyx mori nucleopolyhedrovirus plays a role in budded virus production.

Authors

Guo ZJ, Qiu LH, An SH, Yao Q, Park EY, Chen KP, Zhang CX.

 

Institute of Life Science and School of Finance and Economics, Jiangsu University, 301 Xuefu Road, Zhenjiang 212013, Jiangsu, PR China.

Journal

Virus Res. 2010 May 16. [Epub ahead of print]

Abstract

Open reading frame 60 (bm60) of the Bombyx mori nucleopolyhedrovirus (BmNPV) is a conserved gene among group I and some group II NPVs. bm60 encodes a late expressed protein that localizes to both the cytoplasm and nucleus of infected cells. This paper describes the characterization of a BmNPV mutant (vbm60-Null) lacking functional bm60. It was observed that the production of budded virus (BV) was reduced by nearly an order of magnitude relative to wt virus in vbm60-Null-infected BmN cells and B. mori larvae. Quantitative real-time PCR assay showed that the viral DNA replication was affected in infected cells due to disruption of bm60. Larval bioassays showed that the speed of kill of vbm60-Null virus was greatly reduced, as it took approximately 28-36h longer to kill the fifth instar B. mori larvae. These results suggest that BmNPV bm60 is not essential for viral replication, but required for efficient BV production. Copyright © 2010 Elsevier B.V. All rights reserved.

Citation

PMID: 20576538 [PubMed - as supplied by publisher]

 

 

Title

Characterization of a germination-accelerating factor from the silkworm ( Bombyx mori Linnaeus) of entomopathogenic fungus Nomuraea rileyi (Farlow) Samson. Free Article

Authors

Noda T, Ono M, Iimure K, Araki T.

 

Kumamoto Prefectural Agriculture Research Center, Koshi, Kumamoto, Japan. noda-t-dw@pref.kumamoto.lg.jp

Journal

Biosci Biotechnol Biochem. 2010 Jun 23;74(6):1226-30. Epub 2010 Jun 7.

Abstract

The conidium of the entomopathogenic fungus, Nomuraea rileyi, has been found to germinate rapidly in the presence of a host insect-derived extract. This extract therefore appears to contain an important factor involved in host recognition by N. rileyi, although the substance (germination-accelerating factor, GAF) responsible for such unique germination behavior has yet to be identified. Our previous study was extended to the isolation of GAF from pupae of the silkworm, a host insect of N. rileyi. This present work subjects GAF to a structural analysis. The chemical structure of GAF is characterized as 2S-amino-tetradeca-4-ene-1,3R-diol (D-erythro-C(14)-sphingosine) based on spectroscopic data. An examination of the structure-activity relationship shows that the activity of D-erythro-C(14)-sphingosine was superior to that of sphingosines with shorter and longer carbon chains. It is suggested that the molecular species with a 14-carbon chain of a sphingosine is important for host recognition.

Citation

PMID: 20530914 [PubMed - in process]

 

Title

Isolation of a bioactive substance from the silkworm (Bombyx mor i Linnaeus) that accelerates the germination of the entomopathogenic fungus Nomuraea rileyi (Farlow) Samson. Free Article

Authors

Noda T, Ono M, Iimure K, Araki T.

 

Kumamoto Prefectural Agriculture Research Center, Koshi, Kumamoto, Japan. noda-t-dw@pref.kumamoto.lg.jp

Journal

Biosci Biotechnol Biochem. 2010 Mar 23;74(3):563-8. Epub 2010 Mar 7.

Abstract

The conidium of the entomopathogenic fungus Nomuraea rileyi has been found to germinate rapidly in the presence of host insect-derived extracts. Thus the extract appears to contain an important factor involved in host recognition by N. rileyi. However, the substance responsible for such unique germination behavior has yet to be identified. Hence we attempted to purify this substance. One thousand g of dried silkworm pupae was subjected to methanol extraction, followed by methanolysis, two different solvent partitions, and three different column chromatographies. A total of 12.4 mg of substance was obtained in the active fraction. The substance obtained exhibited an activity more than 46,000 times higher than that of the methanol extract. The substance was detected as a single peak on Sephadex LH20 column chromatography and as a single band on high-performance thin-layer chromatography. These data indicate that the concentrated fraction contained a high-purity substance.

Citation

PMID: 20208348 [PubMed - indexed for MEDLINE]

 

Title

Efficient silkworm expression of single-chain variable fragment antibody against ginsenoside Re using Bombyx mori nucleopolyhedrovirus bacmid DNA system and its application in enzyme-linked immunosorbent assay for quality control of total ginsenosides.

Authors

Sakamoto S, Pongkitwitoon B, Nakamura S, Maenaka K, Tanaka H, Morimoto S.

 

Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.

Journal

J Biochem. 2010 Jun 30. [Epub ahead of print]

Abstract

A single-chain variable fragment (scFv) antibody against ginsenoside Re (G-Re) have been successfully expressed in the silkworm larvae using Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid DNA system. The baculovirus donor vector for expression of scFv against G-Re (GRe-scFv) was constructed to contain honeybee melittin signal sequence (HMSS) to accelerate secretion of the recombinant GRe-scFv into the hemolymph of silkworm larvae. Functional recombinant GRe-scFv was purified by cation exchange chromatography followed by immobilized metal ion affinity chromatography. The yield of purified GRe-scFv was 6.5 mg per 13 silkworm larvae, which is equivalent to 650 mg/l of the hemolymph, exhibiting extremely higher yield than that expressed in Escherichia coli (1.7 mg per liter of culture medium). It was revealed from characterization that GRe-scFv retained similar characteristic of the parental monoclonal antibody (MAb) against G-Re (MAb-4G10), making it possible to develop indirect competitive enzyme-linked immunosorbent assay (icELISA) for quality control of total ginsenosides in various ginsengs. The detectable range for calibration of G-Re by developed icELISA shows 0.05-10 microg/ml. These results clearly suggested that the silkworm expression system is quite useful for the expression of functional scFv that frequently required time- and cost-consuming refolding when it expressed in Escherichia coli.

Citation

PMID: 20592135 [PubMed - as supplied by publisher]

 

Title

Topical formulations of serratiopeptidase: development and pharmacodynamic evaluation.

Authors

Nirale NM, Menon MD.

 

Department of Pharmaceutics, Bombay College of Pharmacy, Kalina, Santacruz (East), Mumbai-400 098, India.

Journal

Indian J Pharm Sci. 2010 Jan;72(1):65-71.

Abstract

Serratiopeptidase, an enzyme derived from Serratia marcescences strain E-15 (ATCC 21074), present in the gut wall of the silk worm possesses anti-inflammatory properties, and can prove to be a suitable alternative to commonly used non steroidal antiinflammatory agents. Being sensitive to gastric degradation, serratiopeptidase is conventionally given orally in the form of enteric coated tablet formulations. Topical formulations of serratiopeptidase would be useful to treat local inflammations and may prove to be more effective compared to non steroidal antiinflammatory agents. The present study investigates the feasibility of developing topical preparations of serratiopeptidase in the form of ointments and gels. Excipient compatibility of serratiopeptidase with various excipients and polymers, formulation development, characterization and stability studies have been carried out. Stable formulation was evaluated for anti-inflammatory activity by oxazolone induced ear edema method in mice and allergenic potential by passive cutaneous anaphylaxis.

Citation

PMCID: PMC2883229, PMID: 20582192 [PubMed - in process]

 

posted Jul 9, 2010, 9:28 AM by rajesh gk

Title

[Study on the biology feature and pathogenicities to silkworm of a microsporidium isolated from Barathra brassicae L.]

[Article in Chinese]

Authors

Zheng X, Zhou Y, Huang B, Lu K, Huang X.

Institute of Sericulture, Guangdong Academy of Agricultural Sciences, Guangzhou 510640.

Journal

Wei Sheng Wu Xue Bao. 2000 Oct;40(5):540-4.

Abstract

A microsporidium (called as Bab-M) was isolated from Barathra brassicae L. captured from suburban vegetable plot of Guang Zhou. The spores were long-ovoid in shape and 4.02 +/- 0.36 microns x 1.99 +/- 0.36 microns in size. Immunologically the microsporidium shared spore surface specific antigen(s) with N. bombycis. The ultrastructure and life cycle of Bab-M were similar to that of N. b.. The rate of transovarian transmission was high. The initial conclusion was that Bab-M should be referred to as Nosema bombycis, but there was variation between them.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/12548768 PMID: 12548768 [PubMed - indexed for MEDLINE]

 

Title

Immune stimulation in the silkworm, Bombyx mori L., by CpG oligodeoxynucleotides.

Authors

Kim I, Kim SH, Lee YS, Yun EK, Lee HS, Kim JW, Ryu KS, Kang PD, Lee IH.

Department of Sericulture & Entomology, National Institute of Agricultural Science and Technology, Rural Development Administration, Suwon, South Korea.

Journal

Arch Insect Biochem Physiol. 2004 Jan;55(1):43-8.

Abstract

Synthetic ODNs containing unmethylated CpG dinucleotides are known to stimulate immune responses in vertebrates, but so far the effect has not been studied in insects. In this report, we describe an induction of immune response following injection of oligodeoxynucleotides (ODNs) into the insect hemocoel. The fifth instar silkworm (Bombyx mori L.) larvae were injected with several synthetic ODNs containing variable number of unmethylated CpG motifs, heat-denatured genomic DNA of B. mori itself, or intact genomic DNA to observe a new induction pattern in the insect immune mechanism. When the induction of immune response was examined based on the expression rates of genes for antibacterial peptides such as attacin and cecropin, we could confirm that it was triggered upon injection of ODNs. The expression was, however, neither dependent on numbers of CpG motifs nor methylation of CpGs in ODNs. Furthermore, it was confirmed that the presence of CpG in ODN was not involved in the induction pattern of insect immunity caused by ODNs, although it has been reported that vertebrates respond in a specific manner against invading ODNs containing CpG dinucleotides. In addition, insect immunity was not stimulated by injection of intact DNA from host. In contrast, the injection of denatured genomic DNA provoked the host immune reaction. Taken together, our data suggest that foreignness of ODNs or DNA might be a key factor in the induction of insect immunity. Copyright 2003 Wiley-Liss, Inc.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/14691962 PMID: 14691962 [PubMed - indexed for MEDLINE]

 

Title

Characterization of Antheraea pernyi nucleopolyhedrovirus p11 gene, a homologue of Autographa californica nucleopolyhedrovirus orf108.

Authors

Shi SL, Pan MH, Lu C.

Key Sericultural Laboratory of Agriculture Ministry, College of Sericulture and Biotechnology, Southwest University, Chongqing, 400716, China.

Journal

Virus Genes. 2007 Aug;35(1):97-101. Epub 2006 Oct 27.

Abstract

Antheraea pernyi nucleopolyhedrovirus (ApNPV) p11 gene is 309 bp long, potentially encoding 102 amino acids with a predicted molecular weight of 11.2 kDa. ApNPV p11 gene was cloned into the prokaryotic expression vector pQE-30 and P11 was expressed in E. coli M15. Polyclonal antiserum was made against 6xHis tagged P11 protein expressed in E. coli M15. P11 gene transcription was detected as early as 36 h post-infection (p.i.) in tussah pupa and remain at high level up to 96 h p.i. Structural localization revealed that P11 protein was present in polyhedral inclusion bodies (PIB) dilute alkaline saline (DAS) pellet (P) fractions and occlusion-derived virus (ODV), but not in PIB DAS supernatant (S) fractions and budded virus (BV). These results indicated that P11 was associated with ApNPV structure.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/17072759 PMID: 17072759 [PubMed - indexed for MEDLINE]

 

Title

Effects of Wolbachia in the uzifly, Exorista sorbillans, a parasitoid of the silkworm, Bombyx mori. Free PMC Article

Authors

Puttaraju HP, Prakash BM.

Seribiotechnology Research Laboratory, Department of Sericulture, Bangalore University, Bangalore-560 056, India. puttarajuhp@hotmail.com

Journal

J Insect Sci. 2005 Nov 11;5:30.

Abstract

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.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/17119612 PMID: 17119612 [PubMed - indexed for MEDLINE]PMCID: PMC1615237

 

Title

[Identification of white muscardin silkworms by infrared spectroscopy]

[Article in Chinese]

Authors

Ji XL, Gai YP, Mu ZM, Wang GX, Ji SL.

Department of Sericulture, Shandong Agricultural University, Taian 271018, China.

Journal

Guang Pu Xue Yu Guang Pu Fen Xi. 2007 Jan;27(1):66-9.

Abstract

The infrared spectra of the ethanol extracts of well-living silkworms and white muscardin silkworms of different seasons and breeds were analyzed by means of the sequential analysis in which two indexes, i. e. common peak ratio and variant peak ratio, were applied. The results showed that the ethanol extracts of white muscardin silkworm have a stable and distinct infrared spectrum. The spectral differences of the ethanol extracts between white muscardin silkworms and well-living silkworms were so obvious that the common peak ratio of them was no more than 63. 0%, and the variant peak ratio amounted to 41. 2%. The spectra of different breeds and seasons conformed with each other with a few small differences. The minimum common peak ratio of the spectra of different breeds was 76. 0%, and the maximal ratio was 92. 0%. The common peak ratio of the spectra of different seasons was 73. 1%. Infrared spectrometry was proved to be good for the identification of white muscardin silkworms and the differentiation of white muscardin silkworms of different breeds and seasons.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/17390651 PMID: 17390651 [PubMed - indexed for MEDLINE

 

Title

A proteomic-based approach for the characterization of some major structural proteins involved in host-parasite relationships from the silkworm parasite Nosema bombycis (Microsporidia).

Authors

Wang JY, Chambon C, Lu CD, Huang KW, Vivarès CP, Texier C.

Equipe Parasitologie Moléculaire et Cellulaire, LBP, UMR CNRS 6023, Université Blaise Pascal, Aubière, France.

Journal

Proteomics. 2007 May;7(9):1461-72.

Abstract

Nosema bombycis is the causative agent of the silkworm Bombyx mori pebrine disease which inflicts severe worldwide economical losses in sericulture. Little is known about host-parasite interactions at the molecular level for this spore-forming obligate intracellular parasite which belongs to the fungi-related Microsporidia phylum. Major microsporidian structural proteins from the spore wall (SW) and the polar tube (PT) are known to be involved in host invasion. We developed a proteomic-based approach to identify few N. bombycis proteins belonging to these cell structures. Protein extraction protocols were optimized and four N. bombycis spore protein extracts were compared by SDS-PAGE and 2-DE to establish complementary proteomic profiles. Three proteins were shown to be located at the parasite SW. Moreover, 17 polyclonal antibodies were raised against major N. bombycis proteins from all extracts, and three spots were shown to correspond to polar tube proteins (PTPs) by immunofluorescent assay and transmission electron microscopy immunocytochemistry on cryosections. Specific patterns for each PTP were obtained by MALDI-TOF-MS and MS/MS. Peptide sequence tags were deduced by de novo sequencing using Peaks Online and DeNovoX, then evaluated by MASCOT and SEQUEST searches. Identification parameters were higher than false-positive hits, strengthening our strategy that could be enlarged to a nongenomic context.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/17407187 PMID: 17407187 [PubMed - indexed for MEDLINE]

 

Title

Frequency of infection with A and B supergroup Wolbachia in insects and pests associated with mulberry and silkworm. Free Article

Authors

Prakash BM, Puttaraju HP.

Laboratory of Seribiotechnology, Department of Sericulture and Life Sciences, Bangalore University, Bangalore 560 056, India.

Journal

J Biosci. 2007 Jun;32(4):671-6.

Abstract

Wolbachia is a ubiquitous, Gram-negative,vertically transmitted, alpha-proteobacterium that causes an array of reproductive abnormalities including cytoplasmic incompatibility, feminization of genetic males, parthenogenesis in a number of insect species, among others. Wolbachia is now being exploited as an agent for pest and vector control. Previous surveys indicated that it is commonly seen in 16-76% of arthropods. In this paper, using polymerase chain reaction assay based on specific amplification of the ftsZ -A and -B supergroup Wolbachia gene fragments, we found that 30% of insects and pests screened were positive for Wolbachia. Among them 66.7% harbour double Wolbachia infection, while 33.3 % harbour single Wolbachia infection. These results indicate widespread infection with both double and single Wolbachia, and provide a wealth of information to exploit this endobacterium for the management of pests and vectors.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/17762140 PMID: 17762140 [PubMed - indexed for MEDLINE]

 

Title

[The analysis of bro genes of GD isolate of Bombyx mori nucleopolyhedrovirus]

[Article in Chinese]

Authors

Pang M, Pan GQ, Li T, Wang X, Zhou ZY.

The Key Laboratory of Sericulture of Agricultural Ministry, Southwest University, Chongqing 400715, China.

Journal

Bing Du Xue Bao. 2007 Nov;23(6):485-9.

Abstract

BmNPV GD isolate from China was plaque-purified and four bro genes were cloned termed as bro-a,b,c,d. The obtained sequences were aligned to the related sequences in GenBank and the BmNPV CQ1 isolate preserved in our laboratory. Compared with genome sequences of BmNPV T3 isolate, bro genes of GD isolate housed insertion and deletion, and the changes of amino acid mainly occured at the N terminal of corresponding protein. The phylogenetic analysis of bro genes indicated that GD bro-d gene belongs to subgroup A together with T3, CQ1 bro-d and SC7 bro- III; GD bro-a, c genes belong to subgroup B together with T3, CQ1 bro-a, c and SC7 bro-II; GD bro-b gene belongs to subgroup C together with T3, CQ1 bro-b, e and SC7 bro-I. The evolutionary relationship of bro genes showed vague relevance to their geographical location. The distribution character of bro genes in four BmNPV isolates is coincidence with the KANG's theory that the bro-d plays an irreplaceable functional role(s) during viral replication, while bro-a and bro-c functionally complement each other. Meanwhile, we postulate that 3 bro genes in SC7 isolate is probable the most simple form of bro genes.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/18092688 PMID: 18092688 [PubMed - indexed for MEDLINE]

 

Title

Identification of a novel spore wall protein (SWP26) from microsporidia Nosema bombycis.

Authors

Li Y, Wu Z, Pan G, He W, Zhang R, Hu J, Zhou Z.

The Key Sericultural Laboratory of Agricultural Ministry, Southwest University, Beibei, Chongqing 400716, PR China.

Journal

Int J Parasitol. 2009 Mar;39(4):391-8. Epub 2008 Sep 27.

Abstract

Microsporidia are obligate intracellular parasites related to fungi with resistant spores against various environmental stresses. The rigid spore walls of these organisms are composed of two major layers, which are the exospore and the endospore. Two spore wall proteins (the endosporal protein-SWP30 and the exosporal protein-SWP32) have been previously identified in Nosema bombycis. In this study, using the MALDI-TOF-MS technique, we have characterised a new 25.7-kDa spore wall protein (SWP26) recognised by monoclonal antibody 2G10. SWP26 is predicted to have a signal peptide, four potential N-glycosylation sites, and a C-terminal heparin-binding motif (HBM) which is known to interact with extracellular glycosaminoglycans. By using a host cell binding assay, recombinant SWP26 protein (rSWP26) can inhibit spore adherence by 10%, resulting in decreased host cell infection. In contrast, the mutant rSWP26 (rDeltaSWP26, without HBM) was not effective in inhibiting spore adherence. Immuno-electron microscopy revealed that this protein was expressed largely in endospore and plasma membrane during endospore development, but sparsely distributed in the exospore of mature spores. The present results suggest that SWP26 is a microsporidia cell wall protein that is involved in endospore formation, host cell adherence and infection in vitro. Moreover, SWP26 could be used as a good prospective target for diagnostic research and drug design in controlling the silkworm, Bombyx mori, pebrine disease in sericulture.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/18854188 PMID: 18854188 [PubMed - indexed for MEDLINE]

 

Title

Protein profile of Nomuraea rileyi spore isolated from infected silkworm.

Authors

Qin L, Liu X, Li J, Chen H, Yao Q, Yang Z, Wang L, Chen K.

Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang 212013, Jiangsu Province, People's Republic of China. qinlvgao@163.com

Journal

Curr Microbiol. 2009 Jun;58(6):578-85. Epub 2009 Mar 14.

Abstract

Nomuraea rileyi (N. rileyi) is the causative agent of the silkworm, Bombyx mori, green muscardine which can cause severe worldwide economical loss in sericulture. Little is known about N. rileyi at the protein level for this entomopathogenic parasite which belongs to the Ascomycota. Here, we employed proteomic-based approach to identify proteins of N. rileyi spores collected from the dead silkworm. In all, 252 proteins were separated by two-dimensional gel electrophoresis (2-DE), and were subjected to mass spectrometry (MS) analysis, 121 proteins have good MS signal, and 24 of them were identified due to unavailability of genomic information from N. rileyi. This data will be helpful in understanding the biochemistry of N. rileyi.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/19288155 PMID: 19288155 [PubMed - indexed for MEDLINE]

 

Title

Molecular analysis of divergence in tachinid Uzi (Exorista sorbillans) populations in India.

Authors

Chatterjee SN, Taraphdar T, Mohandas TP.

SeriBiotech Laboratory, Central Silk Board, Kodathi Campus, Sarjapur Road, Carmelram, 560035 Bangalore, Karnataka, India. shankerc99@yahoo.com

Journal

Genetica. 2005 Sep;125(1):1-15.

Abstract

Exorista sorbillans is a tachinid endoparasitoid of silkworm, Bombyx mori, and is globally known as uzi. It causes economic injury to the cocoon crop in silkworm cultivating areas of India, except those above 400 m above mean sea level (AMSL) in the foothills of the Himalayas (Darjeeling). It is reported that the sericulture tract of south India became infected with this pest only since 1980 through an accidental transportation of cocoons from West Bengal. To ascertain whether the genome of this parasitoid is differentiating into discrete gene pools in contrasting geo-climatic conditions, molecular profiling of four populations (Es (Annatapur), Es(Ramanagaram), Es (Channapatna) and Es(Kodathi) from south India and Es(Murshidabad) from Murshidabad, West Bengal was undertaken with 13 ISSR, 3 RAPD and six non-random primers designed from various repeat sequences of B. mori . MANOVA indicated significance for the Roy's largest root estimate (55.4; F =18.47; p = 0.002) for the variability contributed by the replication. Further, hierarchical clustering done on the basis of Euclidean distance matrix and Nei's unbiased Phylip clustering put Es(Murshidabad) at the maximum distance from those of south India and 29 markers could also be identified which significantly differentiateEs(Murshidabad) from others. However, Nei's statistics for gene diversity in sub-populations reveal considerably high gene-flow (3.44 and 2.51) among the populations around Bangalore. The gene-flow between Es(Murshidabad) and other population is lowest but cannot be ignored. The comparison of endosymbiont specific 16SrRNA and fts Z gene (partial) sequences through clustalW (gcgMSF) revealed a closer relationship of Es(Murshidabad) with Es(Annatapur) and Es (Ramanagaram) and is not congruent with the relationships discussed above. The significance of this maiden study with a tachinid fly-pest is discussed in the context of understanding the diversification of Uzi fly-pest and also establishing this pest as a relevant biological material for studying microevolution in future.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/16175450 PMID: 16175450 [PubMed - indexed for MEDLINE]

 

Title

Gene organization and complete sequence of the Hyphantria cunea nucleopolyhedrovirus genome. Free Article

Authors

Ikeda M, Shikata M, Shirata N, Chaeychomsri S, Kobayashi M.

 

Laboratory of Sericulture and Entomoresources, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan.

Journal

J Gen Virol. 2006 Sep;87(Pt 9):2549-62.

Abstract

The whole-genome sequence of the Hyphantria cunea nucleopolyhedrovirus (HycuNPV) was analysed. The entire nucleotide sequence of the HycuNPV genome was 132 959 bp long, with a G+C content of 45.1 mol%. A total of 148 open reading frames (ORFs) consisting of more than 50 aa were encoded by the genome. HycuNPV shares more than 122 ORFs with other lepidopteran group I NPVs, including Autographa californica MNPV, Bombyx mori NPV, Choristoneura fumiferana MNPV (CfMNPV), Choristoneura fumiferana defective NPV, Epiphyas postvittana MNPV and Orgyia pseudotsugata MNPV (OpMNPV). Six ORFs are identified as being unique to HycuNPV. Most of the HycuNPV ORFs showed higher similarity to CfMNPV and OpMNPV ORFs than to those of the other group I NPVs. HycuNPV encodes two conotoxin-like homologues (ctls), which are observed only in OpMNPV in group I NPVs. HycuNPV encodes three inhibitors of apoptosis (iaps), hycu-iap-1, hycu-iap-2 and hycu-iap-3, a feature that it shares only with CfMNPV. In addition, six homologous regions (hrs) are identified in the HycuNPV genome. These hrs are located in regions similar to those of the OpMNPV hrs, but different from most of the CfMNPV hrs. Based on the close phylogenetic relationship and conservation of group I NPV-specific genes, such as gp64, ie-2 and ptp-1, it is concluded that HycuNPV belongs to the group I NPVs and is most similar to CfMNPV or OpMNPV.

PMID: 16894193 [PubMed - indexed for MEDLINE]

Citation

http://www.ncbi.nlm.nih.gov/pubmed/16894193

 

Title

Inactivation and mechanisms of chlorine dioxide on Nosema bombycis.

Authors

Wang Z, Liao F, Lin J, Li W, Zhong Y, Tan P, Huang Z.

 

Department of Sericulture Science, South China Agricultural University, Guangzhou 510642, China.

Journal

J Invertebr Pathol. 2010 Jun;104(2):134-9. Epub 2009 Dec 29.

Abstract

Biological tests demonstrated that the inactivation of Nosema bombycis (N. bombycis) spores by chlorine dioxide (ClO(2)) occurs very fast and is highly sensitive. The lowest effective inactivation dosage and time was 15mg/mL for 30min. The inactivation of spores was additionally verified by using double color fluorescence stain and spore germination testing. A series of biological changes, including a large number of substrates that were leaked out from the spores included proteins, DNA, polysaccharide, K(+), and Ca(2+), occurred a short time after N. bombycis spores were treated with ClO(2). In addition, the lipid of spores was disrupted and ATPase activity was inhibited, which resulted in the destruction of the inner structure of the spores.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/20036671 PMID: 20036671 [PubMed - in process]

 

Title

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

Authors

Huang L, Cheng T, Xu P, Cheng D, Fang T, Xia Q.

 

Institute of Sericulture and Systems Biology, Southwest University, Chongqing, China.

Journal

PLoS One. 2009 Dec 1;4(12):e8098.

Abstract

Host-pathogen interactions are complex relationships, and a central challenge is to reveal the interactions between pathogens and their hosts. Bacillus bombysepticus (Bb) which can produces spores and parasporal crystals was firstly separated from the corpses of the infected silkworms (Bombyx mori). Bb naturally infects the silkworm can cause an acute fuliginosa septicaemia and kill the silkworm larvae generally within one day in the hot and humid season. Bb pathogen of the silkworm can be used for investigating the host responses after the infection. Gene expression profiling during four time-points of silkworm whole larvae after Bb infection was performed to gain insight into the mechanism of Bb-associated host whole body effect. Genome-wide survey of the host genes demonstrated many genes and pathways modulated after the infection. GO analysis of the induced genes indicated that their functions could be divided into 14 categories. KEGG pathway analysis identified that six types of basal metabolic pathway were regulated, including genetic information processing and transcription, carbohydrate metabolism, amino acid and nitrogen metabolism, nucleotide metabolism, metabolism of cofactors and vitamins, and xenobiotic biodegradation and metabolism. Similar to Bacillus thuringiensis (Bt), Bb can also induce a silkworm poisoning-related response. In this process, genes encoding midgut peritrophic membrane proteins, aminopeptidase N receptors and sodium/calcium exchange protein showed modulation. For the first time, we found that Bb induced a lot of genes involved in juvenile hormone synthesis and metabolism pathway upregulated. Bb also triggered the host immune responses, including cellular immune response and serine protease cascade melanization response. Real time PCR analysis showed that Bb can induce the silkworm systemic immune response, mainly by the Toll pathway. Anti-microorganism peptides (AMPs), including of Attacin, Lebocin, Enbocin, Gloverin and Moricin families, were upregulated at 24 hours post the infection.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/19956592

PMID: 19956592 [PubMed - indexed for MEDLINE]PMCID: PMC2780328

 

Title

Natural selection maintains the transcribed LTR retrotransposons in Nosema bombycis.

Authors

Xiang H, Pan G, Zhang R, Xu J, Li T, Li W, Zhou Z, Xiang Z.

 

Institute of Sericulture and Systems Biology, Southwest University, Beibei, Chongqing 400715, China.

Journal

J Genet Genomics. 2010 May;37(5):305-14.

Abstract

Eight intact LTR retrotransposons (Nbr1-Nbr8) have been previously characterized from the genome of Nosema bombycis, a eukaryotic parasite with a compact and reduced genome. Here we describe six novel transcribed Nbr elements (Nbr9-Nbr14) identified through either cDNA library or RT-PCR. Like previously determined ones, all of them belong to the Ty3/Gypsy superfamily. Retrotransposon diversity and incomplete domains with insertions (Nbr12), deletions (Nbr11) and in-frame stop codons in coding regions (Nbr9) were detected, suggesting that both defective and loss events of LTR retrotransposon have happened in N. bombycis genome. Analysis of selection showed that strong purifying selection acts on all elements except Nbr11. This implies that selective pressure keeps both these Nbrs and their functions in genome. Interestingly, Nbr11 is under positive selection and some positively selected codons were identified, indicating that new functionality might have evolved in the Nbr11 retrotransposon. Unlike other transposable elements, Nbr11 has integrated into a conserved syntenic block and probably resulted in the inversion of both flanking regions. This demonstrates that transposable element is an important factor for the reshuffling and evolution of their host genomes, and may be maintained under natural selection. (c) 2010 Institute of Genetics and Developmental Biology and the Genetics Society of China. Published by Elsevier Ltd. All rights reserved.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/20513631 PMID: 20513631 [PubMed - in process]

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