• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1485-1490
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• 2006

In this study, comparative replicational properties of Hyphantria cunea nucleopolyhedrovirus (HycuNPV) in Lymantria dispar (IPLB-LdElta) and Spodoptera frugiperda (IPLB-Sf21) cell lines were investigated. Our microscopic observations showed that cytopathic effects (CPEs) in LdElta cells appeared 12 h later than those in Sf21 cells. Whereas polyhedral inclusion bodies (PIBs) formed at 48 h postinfection (p.i.) in LdElta cells, it formed at 36 h p.i. in Sf21 cells. Extracellular virus production determined according to the 50% tissue culture infective dose ($TCID_{50}$) method in LdElta cells started about 12 h later when compared with Sf21 cells. Titers of extracellular virus in LdElta and Sf21 cells were calculated as $1.77{\times}10^9$ plaque forming units (PFU)/ml and $5.6{\times}10^9PFU/ml$, respectively, at 72 h p.i. We also showed that viral DNA replication began at 12 h p.i. in both cell lines. Viral protein synthesis was determined by SDS-polyacrylamide gel electrophoresis (PAGE) and polyhedrin synthesis was observed at 12 h p.i. in both cell lines. The results indicate that while the synthesis of macromolecules is 12 h later and production of extracellular virus is almost 3-fold lower in LdElta cells compared with those in Sf21 cells, the LdElta cell line is still a productive cell line for infection of HycuNPV.

• Journal of Microbiology
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• v.41 no.4
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• pp.306-311
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• 2003

In Korea, a Lymantria dispar multinucleocapsid nucleopolyhedrovirus, LdMNPV-NM, was isolated and characterized from dead L. dispar larvae. The polyhedra of LdMNPV-NM were irregularly shaped with a diameter of $1.62{\pm}0.33{\mu}m$. Numerous virions comprised of the multinucleocapsid were evident in the electron microscopic examination of the polyhedra cross sections. These polyhedra were composed of a major protein of 30 kDa. The restriction enzyme digestion patterns of LdMNPV-NM showed that this isolate had some different fragments from those of the Gypchek LdMNPV isolate, although their overall profiles were similar. The deduced amino acid sequence of the enhancin gene of LdMNPV-NM showed differences when compared to previously reported enhancin genes of other LdMNPV strains. These results suggested that the LdMNPV-NM isolate from Korea was a new NPV strain and had a new enhancin gene.

• International Journal of Industrial Entomology and Biomaterials
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• v.19 no.2
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• pp.237-241
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• 2009

The objective of our study was the evaluation of pathogenicity of a local strain of Mamestra brassicae nucleopolyhedrovirus-K1 (MabrNPV-K1) derived from a diseased larva of M. brassicae found in Korea. The effect of temperature and larval instar on the pathogenicity and production of MabrNPV-K1 was determined under laboratory conditions. The median lethal concentration ($LC_{50}$) values of MabrNPV-K1 for 3rd instar larvae were $3.7\times10^4$ PIBs/larva at $20^{\circ}C$, $9.9\times10^2$ PIBs/larva at $25^{\circ}C$ and $3.8\times10^2$ PIBs/larva at $30^{\circ}C$, respectively. The $LC_{50}$ for the 4th instar larvae was similar to that for the 3rd instar larvae. However, the pathogenicity to the 3rd instar larvae was higher than that to the 4th instar larvae. The median lethal time ($LT_{50}$) values of MabrNPV-K1 were 11.4 to 5.0 days at $30^{\circ}C$ and 18.3 to 5.5 days at $25^{\circ}C$ for the 3rd instar larvae. The $LT_{50}$ value was lowered as temperature went up to $30^{\circ}C$ and dependent on viral concentration. In production efficiency of MabrNPV-K1 using M. brassicae larvae, the mortality of the 3rd instar larvae was 100% when inoculated with $1.0\times10^5$ PIBs/larva and the yield of MabrNPV-K1 was maximal. Regarding the mortality, yield of polyhedra, inoculation doses and required time, the $1.0\times10^4$PIBs/larva at $30^{\circ}C$ was determined as optimal conditions producing polyhedra efficiently.

• International Journal of Industrial Entomology and Biomaterials
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• v.17 no.1
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• pp.125-129
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• 2008

The purpose of this study was to investigate the characteristics of Mamestra brassicae nucleopolyhedrovirus (MabrNPV)-K1 isolated in Korea. Polyhedra of MabrNPV-K1 showed irregular appearance in shape with the average diameter $1.8{\mu}m$. MabrNPV-K1 contained a number of nucleocapsids within a viral envelope embedded in polyhedron. The polyhedrin of MabrNPV-K1 was composed of single polypeptide with a M.W. of approximate 31 kDa which is identical to the commercialized MabrNPV, Mamestrin, as a biological control agent. The nucleotide and amino acid sequences within the coding region of MabrNPV-K1 polyhedrin shared 99.0% similarity with the polyhedrin gene from previous reported MabrNPVs. The median lethal concentrations ($LC_{50}$) of MabrNPV-K1 and Mamestrin to M. brassicae larvae were $3.9{\times}10^3$ PIBs/larva and $6.0{\times}10^4$ PIBs/larva, respectively. Mortality of the MabrNPV-K1 against to the third instars larvae was 15 times higher than that of the Mamestrin. The median lethal times ($LT_{50})$ of MabrNPV-K1 by the concentration of polyhedra were lower ($4.4{\sim}6.1$ days) than those of Mamestrin ($4.1{\sim}8.6$ days). These results suggest that a local strain MabrNPV-K1 has high pathogenicity to M. brassicae and may be useful for the development of biological control agent to control this.

• Microbiology and Biotechnology Letters
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• v.29 no.1
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• pp.18-24
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• 2001

Expression of the baculovirus major envelope glycoprotein gene(gp64) is regulated by transcription from botha early and late promoters. To develop a transient expression vector under the control of gp64 gene promoter, the gp64 gene of Bombyx mori nucleopolyhedrovirus-K1(BmNPV-K1) was characterized. The gp64 gene was local-ized at EcoR I-Pst I 7.38-kb fragment of the BmNPV-K1 genome. The EcorR 1-Pst I 7.38-kb fragment was cloned and the nucleotide sequence of 2,277 bases including the coding region of gp64 gene was determined. Based on these results, transient expression vector using gp64 gene promoter was constructed and named as pBm64. E.coli lacZ gene was introduced onto pBm64 as a reporter gene and expressed transiently in B. mori 5(Bm 5) cells. The expression vector transfected into the cells was maintained stably for 1 to 5 days. In order to confirm the expression of the reporter gene by gp64 promoter, recombinant virus was constructed. The recombinant virus has two independent transcription units in opposite orientations with two promoters; gp64 and polyhedrin gene promoters each initiating transcription of $\beta$-galactosidase and polyhedrin, respectively. Polyhedra formation and expression of $\beta$-galactosidase in Bm5 cells infected with the recombinant virus were observed with phase contrast microscope and in situ staining.

• Korean journal of applied entomology
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• v.43 no.4 s.137
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• pp.329-332
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• 2004

This experiment was conducted to investigate pathogenicity of Spodoptera exigua nucleopolyhedrovirus (SeNPV) with different temperatures for mass production. In laboratory condition, $LC_{50}$ values of SeNPV were $9.797{\times}10^{3}PIBs/mL$ at $20^{\circ}C$ and $3.351{\times}10^{2}PIBs/mL$ at $32^{\circ}C$ in 2nd instar larvae. $LC_{50}$ of the other larval stage were similar to that of 2nd instar. $LT_{50}$ values of SeNPV was 9.0 days in $1.0{\times}10^{3}PIBs/mL$ but 6.9 to 3.5 days in $1.0{\times}10^{4-6}PIBs/mL$ against 3rd instar of Spodoptera exigua. $LT_{50}$ Values of $1.0{\times}10^{4}PIBs/mL$ were 5.7, 5.5 and 4.9 days in 24, 28 and $32^{\circ}C$, respectively. As a results, $LT_{50}$ was shortened with increase of temperatures up to $32^{\circ}C$ and also dependent on viral concentration and larval instars.

• Journal of Sericultural and Entomological Science
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• v.50 no.2
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• pp.53-62
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• 2012

Silkworm larvae often suffer from viral infections causing heavy losses to the economy of silk industry. Insects exhibit both humoral and cellular immune responses that are effective against various pathohens like bacteria, fungi, protozoa, etc., but no insect immune responses is effective against viral infection. To obtain genes related to insect antiviral immunity from Bombyx mori, the cDNA library was constructed from B. mori nucleopolyhedrovirus (BmNPV)-infected B. mori. From the cDNA library, we selected 411 differentially expressed clones, and the 5' ends of the inserts were sequenced to generate ESTs. In this work, 135 unigenes were generated after the assembly of 411 differentially expressed clones ESTs. Of these 135 unigenes, we selected 109 antiviral response-related candidates except 26 clones that high similarity with genes derived from BmNPV. Among 109 unigenes, a total of 80% had significant matches to genes from other organisms in the database, wheres 20% of the unigenes had not matched in the database. Functional groups of these sequences with matches in database were constructed according to their putative biological function. Three largest categories were control of cellular oraganization (52%), metabolism (20%), and protein fate (10%). The genetic information reported in this study will provide more information about antiviral-related genes in silkworms.

• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2001.05a
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• pp.110-110
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• 2001

• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2004.05a
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• pp.172-172
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• 2004

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2001.04a
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• pp.31-31
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• 2001

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