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

作　　者：Claudia Regina das Neves Saez Roxelle Ethienne Ferreira Munhoz Naiara Climas Pereira Thais Souto Bignotto Veronica Aureliana Fassina Graziele Milani Pessini Laura Beatriz Garay Lucineia de Fatima Chasko Ribeiro Rose Meire Costa Brancalhao Maria Aparecida Fernandez 

机构地区：[1]Departamento de Biotecnologia,Genetica e Biologia Celular,Universidade Estadual de Maringa,Maringa,Brasil [2]Seda y Fibras,Hernandarias,Paraguay [3]Universidade Estadual do Oeste do Parana,Cascavel,Brasil

出　　处：《Open Journal of Genetics》2014年第5期370-377,共8页遗传学期刊（英文）

基　　金：supported by CAPES,CNPq,FINEP/Fundacao Araucaria and Secretaria de Estado da Ciencia,Tecnologia e Ensino Superior—FUNDO PARANA.

摘　　要：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.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.

关 键 词：Silkworm Diseases SERICULTURE Viruses in Insects BMNPV Bombyx mori 

分 类 号：R73[医药卫生—肿瘤]