跳至主要内容

Detection of Contamination and Analysis of Vertical Transmission of BmNPV in Eggs and Moths of Bombyx mori

Read full paper at:
http://www.scirp.org/journal/PaperInformation.aspx?PaperID=48990#.VJeFacCAM4

This study reports the molecular detection of Bombyx mori nucleopolyhedrovirus (BmNPV) in silkworm strains of the Universidade Estadual de Maringá Brazilian Germplasm Bank (UBGB). DNA extraction was carried out by using six Bombyx mori female moths of each strain, followed by PCR amplification. A pair of primers was designed based on a specific sequence of the baculovirus genome related to the BmNPV ORF 14. Another pair of primers was used to amplify the silkworm Actin A3 gene segment, which was used as positive control. Twenty gene pools were analyzed, and fifteen revealed a fragment of 443 base pairs (bp), which indicated the presence of the BmNPV. The frequency of contaminated moths was as following: 100% for silkworm strains M18-2, M12-2 and J1; 83% for C25, C75 and C24 strains; 66% for KR01; 50% for M11-A; 33% for AS3, B106, M8 and M11 and 16% for C211, E8 and Hindu strains. These are promising results for the identification of contaminated B. mori moths by BmNPV, which may prevent virus proliferation in subsequent generations. We also analyzed DNA samples extracted from B. mori eggs, but the results were not conclusive regarding the detection of the fragments of the expected size (443 bp). The difficulty in detecting BmNPV contamination in B. mori eggs may be due to the low concentration of virus in samples.
Cite this paper
Saez, C. , Munhoz, R. , Pereira, N. , Bignotto, T. , Fassina, V. , Pessini, G. , Garay, L. , Ribeiro, L. , Brancalhão, R. and Fernandez, M. (2014) Detection of Contamination and Analysis of Vertical Transmission of BmNPV in Eggs and Moths of Bombyx mori. Open Journal of Genetics, 4, 370-377. doi: 10.4236/ojgen.2014.45034
 

[1] Hanada, Y. and Watanabe, J.K. (1986) Manual de criação do bicho-da-seda. Cocamar, Curitiba.
[2] Nagaraju, J. and Goldsmith, M.R. (2002) Silkworm Genomics—Progress and Prospects. Current Science, 83, 411-425.
[3] Yamamoto, T. (2000) Silkworm Strains. In: Nakatsuij, N., Ed., Strains Maintenance and Databank for Life Science, Kyoritsu Shuppan, Tokyo, 45-49.
[4] Zanetti, R. (2005) Doenças e pragas do bicho-da-seda, notas de aula de ENT 100—Sericicultura.
http://www.den.ufla.br/siteantigo/Professores/Ronald/Disciplinas/Notas%20Aula/Sericicultura%
20doencasbicho.pdf
[5] Sengupta, K., Kumar, P. and Baig, M. (1990) Handbook on Pest and Disease Control of Mulberry and Silkworm. Economic and Social Commission for Asia and Pacific, UNESCAP (United Nations Economic and Social Commission for Asia and the Pacific), Bangkok.
[6] Brancalhão, R.M.C. (2002) Vírus entomopatogênicos no bicho-da-seda: Taxonomia e citopatologia causada por nucleopolyhedrovirus em células de Bombyxmori. Biotecnologia Ciência e Desenvolvimento, 24, 54-58.
[7] Jehle, J.A., Blissard, G.W., Bonning, B.C., Cory, J.S., Herniou, E.A., Rohrmann, G.F., Theilmann, D.A., Thiem, S.M. and Vlak, J.M. (2006) On the Classification and Nomenclature of Baculoviruses: A Proposal for Revision. Archives of Virology, 151, 1257-1266.
http://dx.doi.org/10.1007/s00705-006-0763-6
[8] Beukema, S.J. (1992) Towards More Realistic Models of Disease and Insect Population Dynamics: Virus-Tent Caterpillar Interactions. Doctoral Thesis, University of British Columbia, Vancouver.
[9] Neelgund, Y.F. and Mathad, S.B. (1978) Transmission of Nuclear Polyhedrosis Virus in Laboratory Population of the Armyworm, Mythimna (Pseudaletia) separata. Journal of Invertebrate Pathology, 31, 143-147.
http://dx.doi.org/10.1016/0022-2011(78)90001-0
[10] Ridhards, A., Matthews, M. and Christain, P. (1998) Ecological Considerations for the Environmental Impact Evaluation of Recombinant Baculovirus Insecticides. Annual Review of Entomology, 43, 493-517.
http://dx.doi.org/10.1146/annurev.ento.43.1.493
[11] Kukan, B. (1999) Vertical Transmission of Nucleopolyhedrovirus in Insects. Journal of Invertebrate Pathology, 74, 103-111.
http://dx.doi.org/10.1006/jipa.1999.4873
[12] Fuxa, J.R., Weidner, E.H. and Richter, A.R. (1992) Polyhedra without Virions in a Vertically Transmitted Nuclear Polyhedrosis Virus. Journal of Invertebrate Pathology, 60, 53-58.
http://dx.doi.org/10.1016/0022-2011(92)90153-U
[13] Fuxa, J.R., Richter, A.R., Ameen, A.O. and Hammock, B.D. (2002) Vertical Transmission of TnSNPV, TnCPV, AcMNPV and Possibly Recombinant NPV in Trichoplusia ni. Journal of Invertebrate Pathology, 79, 44-50.
http://dx.doi.org/10.1016/S0022-2011(02)00003-4
[14] Ikuno, A.A., Margatho, L.F.F., Harakava, R., Akamatsu, M.A., Martins, E.M.F., Porto, A.J. and Ferreira, V.C.A. (2004) Direct Application of the New PCR Protocol for Evaluation and Monitoring of Bombyx mori Infection by Nucleopolyhedrovirus. Arquivos do Instituto Biologico, 71, 309-315.
[15] Khurad, A.M., Mahulikar, A., Rathod, M.K., Rai, M.M., Kanginakudru, S. and Nagaraju, J. (2004) Vertical Transmission of Nucleopolyhedrovirus in the Silkworm, Bombyx mori L. Journal of Invertebrate Pathology, 87, 8-15.
http://dx.doi.org/10.1016/j.jip.2004.05.008
[16] Khumnoi, S. (2007) Vertical Transmission of Nucleopolyhedrovirus in Thai Mulberry Silkworm, Bombyx mori L. (Lepidoptera: Bombycidae). Master of Science (Agricultural Biotechnology), Kasetsart University, Bangkok.
[17] Pereira, N.C., Munhoz, R.E.F., Bignotto, T.S., Bespalhuk, R., Garay, L.B., Saez, C.R.N., Fassina, V.A., Nembri, A. and Fernandez, M.A. (2013) Biological and Molecular Characterization of Silkworm Strains from the Brazilian Germplasm Bank of Bombyx mori. Genetics and Molecular Research, 12, 2138-2147.
http://dx.doi.org/10.4238/2013.June.28.1
[18] TakahashiI, K., Takaku, M. and Hasegawa, M. (1925) A Experiment of Dissection and Transparent Method with Silkworm Egg. Sanshishinpou, 33, 47-59.
[19] Hatakeyama, Y. and Hayasaka, S. (2003) A New Method of Pebrine Inspection of Silkworm Egg Using Multiprimer PCR. Journal of Invertebrate Pathology, 82, 148-151.
http://dx.doi.org/10.1016/S0022-2011(03)00019-3
[20] Santos, S.A., Silva, J.L.C., Balani, V.A., Seixas, F.A.V. and Fernandez, M.A. (2010) Conserved Baculoviral ORFs 10 and 14 from Bombyx mori Multiple Nucleopolyhedrovirus. Genetics and Molecular Research, 9, 457-470.
http://dx.doi.org/10.4238/vol9-1gmr723
[21] Gomi, S., Majima, K. and Maeda, S. (1999) Sequence Analysis of the Genome of Bombyx mori Nucleopolyhedrovirus. Journal of General Virology, 80, 1323-1337.
[22] Silva Jr., W.A., Costa, M.C., Valente, V., Sousa, J.F., Paço-Larson, M.L., Espreafico, E.M., Camargo, S.S., Monteiro, E., Holanda, A.J., Zago, M.A., Simpson, A.J. and Neto, E.D. (2001) PCR Template Preparation for Capillary DNA Sequencing. Biotechniques, 30, 537-542.
[23] Kaewwises, M., Attathom, T., Chaeychomsri, S. and Chowpongpang, S. (2004) Detection of Nucleopolyhedrovirus of Mulberry Silkworm, Bombyx mori by PCR Technique. Proceedings of the 42nd Kasetsart University Annual Conference, Kasetsart, 3-6 February 2004, 174-182.
[24] Altschul, S.F., Gish, W., Miller, W., Myers, E.W. and Lipman, D.J. (1990) Basic Local Alignment Search Tool. Journal of Molecular Biology, 215, 403-410.
http://dx.doi.org/10.1006/jmbi.1990.9999
[25] Thompson, J.D., Higgins, D.G. and Gibson, T.J. (1994) CLUSTAL W: Improving the Sensitivity of Progressive Multiple Alignment through Sequence Weighting, Positions-Specific Gap Penalties and Weight Matrix Choice. Nucleic Acids Research, 22, 4673-4680.
[26] Kaewwises, M. (2006) Potential Application of PCR-Based Method for Early Detection Osgrasserie Disease of Silkworm Bombyx mori. Doctoral Thesis, Kasetsart University, Bangkok.                           eww141222lx

评论

此博客中的热门博文

Does Immigration Promote the Investment of the Monopolistic Firm?

In the present paper, we examine the effect of increasing uncertainty of immigrants’ growth on the optimal timing of investment of a firm that has a monopolistic power over the labor market. It is revealed that when the uncertainty of immigrants’ growth is more than a threshold level, increasing uncertainty of immigrants’ growth accelerates the optimal timing of firms’ investment and enhances the economic growth, even if the uncertainty of immigrants’ growth is formulated by the geometric Brownian motion, which is in sharp contrast to the standard result that an increase in the uncertainty postpones the optimal timing. With an increase in the immigrants over the past ten years, workforces in the host countries have been growing significantly to the extent that the immigrants represent 70% of the increase in the workforce in Europe, and 47% in the United States as OECD indicates. In the present paper, we attempted to investigate the effect of increased uncertainty caused by the growi...

Education Policy Implementation: A Mechanism for Enhancing Primary Education Development in Zanzibar

Education is one of the fundamental rights of individuals; therefore, the government of a country needs to develop and strengthen educational policy and quality as well as to ensure that everyone has equal access to basic education. The improvement of access and quality of education in the world is becoming as an essential factor in development, whereas the basic education (primary school), is acknowledged as a foundation of the higher educational development for every country. To fulfill this goal, governments introduce several policies and procedures; however, it requires some reforms and participation from the politician, policymakers, and other stakeholders to re-examine educational policy so that it can lead to multiplication and betterment of the reforms. Educational reforms actually focus on accountability. A positive educational development and reform is very challenging and needs more effort and strategy on how to use and utilize the resources effectively as such it can achie...