• International Journal of Industrial Entomology and Biomaterials
• /
• v.6 no.1
• /
• pp.49-55
• /
• 2003

We describe the generation of transgenic silkworm that carrying the chimeric fibroin light chain (L-chain) gene. Previously, we have cloned the complete fibroin L-chain gene from the silkworm Baekok-Jam, Bombyx mori, and the complete fibroin gene from the oak silkworm, Antheraea yamamai. The 444 bp repetitive sequence of A. yamamai fibroin gene was inserted into the exon 6 of B. mori fibroin L-chain gene to produce chimeric fibroin L-chain gene. The chimeric fibroin L-chain gene was cloned into the polyhedrin gene site of Autographa californica nuclear polyhedrosis virus (AcNPV) to yield a recombinant baculovirus as a fibroin gene targeting vector, One-day-old fifth instar female silkworm larvae were injected with the recombinant baculovirus and then mated with normal male moths. Genomic DNA from their progenies was extracted and screened for the desired targeting event by using PCR and Southern blot analysis. The analysis showed that the chimeric fibroin gene had intergrated into the L-chain gene on the genome by homologous recombination and was transmitted through generations. The transgenic silkworm carrying the chimeric fibroin gene were approximately 43.2% in $F_2$ generation, and the silkworms synthesized the fusion protein in cocoons layer.

• Journal of Sericultural and Entomological Science
• /
• v.32 no.2
• /
• pp.105-109
• /
• 1990

The disinfectivity of formalin fumigation was tested against silkworm nuclear polyhedrosis virus and yellow muscardine pathogen, Beauveria bassiana. The inactivation of the virus was acquired when it was fumigated by adding 30g of potassium permanganate to 75$m\ell$ of formalin per 3.3$m^2$ of rearing room area and viability of the yellow muscardine pathogen was also lost with the same treatment of fumigation. It was also proved that the fumigation didn't give and damage to silkworm larvae when it was applied 2 or 3 times to grown larvae.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.4 no.1
• /
• pp.43-49
• /
• 2002

We have cloned cDNAs encoding toxin from the spider, Araneus ventricosus, and constructed a recombinant baculovirus expressing the insecticidal toxin. The cDNAs encoding toxin were cloned from the cDNA library of A. ventricosus. Sequence analysis of the cDNAs encoding the toxin of A. ventricosus revealed that the 240 bp cDNA for AvTox-1 and 192 bp cDNA for AvTox-2 have an open reading frame of 80 and 64 amino acid residues, respectively. The deduced protein sequence of the toxin genes of AvTox-1 and AvTox-2 was aligned to that of the snack Anemonia sulcata and scorpion Centruroides limpidus limpidus, respectively. Northern blot analysis indicated that AvTox-2 toxin gene showed a fat body-spe-cific expression pattern at the transcriptional level. Furthermore, we have explored the possibility of improving baculovirus by incorporating the A. vontricosus toxin gene into Bombyx mori nuclear polyhedrosis virus genome under the control of polyhedrin promoter, The AvTox-2 toxin gene was expressed as approximately 5.8 kDa band in the recombinant baculovirus-injected silkworm larvae. Bioassays with the recombinant virus expressing AvTox-2 on 5th instar silkworm larvae demonstrated a decrease in the time to kill $(LT_{50} days)$ compared to wild-type BmNPV-Kl $(LT_{50} 6.72 days)$ in the injection of 10 viruses. These results indicate that A. ventricosus toxin is a novel member of the spider toxin family, suggesting that the toxin gene can be used in recombinant baculoviruses to reduce insect feeding damage and increase the speed of insect kill.

• Journal of Sericultural and Entomological Science
• /
• v.22 no.2
• /
• pp.8-12
• /
• 1981

Practical application of silkworm artificial diet is very desirable to save labour in sericultural industry as the problem of labour shortage is becoming serious in Korea. However, silkworms reared on the artificial diet are more susceptible to viruses than those reared on mulberry leaves because of the lower anti-viral activity of gut juice of silkworms grown on artificial diet compared with that of silkworms grown on mulberry leaves. In this study, authors investigated the varietal difference of silkworm larvae, Bombyx mori L., reared on artificial diet which contained 20 percent of dried mulberry powder, in the susceptibility to nuclear polyhedrosis virus (NPV). The results showed that there is no difference in susceptibility to NPV among tested varieties when high concentration of NPV was admitted to silkworm larvae, but varietal difference appeared in lower concentration admitted. Among 7 hybrids tested, Hansaeng 1${\times}$Hansaeng 2 was most resistant to NPV with an $LC_{50}$ of 2.7${\times}$10$\^$6/ and Jam 111${\times}$Jam 112 was also more resistant comparatively than other hybrids.

• Journal of Sericultural and Entomological Science
• /
• v.8
• /
• pp.51-62
• /
• 1968

This study was carried out to investigate the life cycle and the damage of Tricholyga sorbillans Wiedmann to silkworms(Bombyx mori L.). The results are summarized as follows. 1. This insect has 5 or 6 generations per year in Suwon area. 2. The egg, larval, and pupal stage are about 2, 8 and 18 days respectively. 3. The weather conditions considerably affect the life cycle, that is, the 3 rd generation is the shortest of all, lasting for 21 days and l4 hours, and the 5th generation is the longest for about 40 days. 4. The larvae come out of silkworm bodies, burrowing into the ground 3.5cm to 10.5cm in depth before changing into the pupae. 5. This insect does not overwinter as pupa, but seems to do as larva, parasiting on the field insects. 6. The damage of Tricholyga sorbillans Wiedmann to silkworm was 27.9％, it is so high that the silk worms could not be reared outside without a reasonable control method. 7. When the full-grown silkworm is parasited by one egg of these insects, there is not only enough possibility of spinning. but also without any effects of their larvae on the cocoon quality. 8. The damage of Tricholyga sorbillans Wiedmann throughout the country covers 6.36％ in spring, 4.09％ in fall, and 5.25％ yearly mean.

• Journal of Sericultural and Entomological Science
• /
• v.36 no.1
• /
• pp.69-75
• /
• 1994

Through the histological and anatomical investigation of silkworm gonad, N. bombycis infection was found to begin from the peripheral region of ovarial sheath or testicular sheath, then, the pathogens spread to the inner portion. Peroral inoculation with purified spores of N. bombycis to 2nd instar larvae at dosages around 106-8/㎤ of artificial diet resulted in the extremely extended larval survival as long as 15 to 22 days of 4th instar. The growth of ovarioles was confirmed in the 10 to 14 day old larvae, oogonia developed into oocytes and nurse cell against heavy infection of the ovary. Gonads rarely obstructed oogenesis and spermatogenesis in the pupae failed in adult eclosion. Light infection of female hosts effected insignificantly on the ovarial development, however, recorded 100% transmission of the pathogens to the progeny populations. Conclusively, ovarial inflection of silkworm induce transovarial transmission begins around 2~3 day old pupae when ovarioles extruded out to hemocoel, and the infection period thought to be continued until the stage of eggs complete shell formation in the ovariole.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.27 no.2
• /
• pp.237-242
• /
• 2013

We investigated the 1-deoxynojirimycin (1-DNJ) content of extracts from silkworm larvae at each developmental stage within three silkworm varieties. We also compared the content of the following polyhydroxylated alkaloids in the silkworm extracts: 1-DNJ, fagomine, and 1,4-dideoxy-1,4-imino-d-arabinitol (DAB). In addition, we evaluated the glucose-lowering effects of silkworm extract powder in db/db mice. The 1-DNJ content was the highest in Yeonnokjam $5^{th}$ instar $3^{rd}$ d larvae and Hansaengjam $5^{th}$ instar $3^{rd}$ d larvae, which contained 18.4 mg/100 g dry weight and 18.3 mg/100 g dry weight, respectively. The $5^{th}$ instar $3^{rd}$ d larvae exhibited a higher content of 1-DNJ than that of $5^{th}$ instar $5^{th}$ d larvae among all varieties. The glucose-lowering effects of silkworm extracts and Yeonnokjam powder were tested on db/db mice, and the blood glucose levels were found to decrease significantly in the YR70 group. Silkworm extracts (180 mg/kg, 90 mg/kg, 45 mg/kg, and 22.5 mg/kg) and acarbose (50 mg/kg) were administered orally for 4 wk. Changes in water intake were not statistically significant between control and silkworm extract-treated groups. Compared to the control group, blood glucose levels in the silkworm extract powder-treated group decreased in the 22.5 mg/kg/d group after being administered for 4 wk. This decrease was statistically significant. Furthermore, biochemical changes in the AST(Aspartate aminotransferase), ALT(Alanine aminotransferase), TCHO(Total Cholesterol), TG(Triglyceride), LDL(Low density lipoprotein), and HDL(High density lipoprotein) levels in blood were not observed. However, statistically significant decreases in blood GLU in the 22.5 mg/kg/d group compared to that of the control group occurred. In addition, the epididymal fat weight of the silkworm extract powder-treated group decreased significantly in both the 22.5 mg/kg/d group and 180 mg/kg/d group compared to that of the control group, but there were no statistically significant changes in perirenal fat weight. These results demonstrate that silkworm extracts inhibit changes in blood glucose levels in model diabetic mice.

• Korean journal of applied entomology
• /
• v.36 no.4
• /
• pp.341-350
• /
• 1997

We have now constructed a novel recombinant baculovirus producing fusion protein with Autographa californica nuclear polyhedrosis virus (AcNPV) polyhedrin and Bacillus thuringiensis(Bt) cryIA(c) crystal protein. The fusion protein expressed by the recombinant baculovirus in insect cells was characterized. The N-terminal of cryIA(c) gene of Bt subsp. kurstaki HD-73 was introduced under the control of polyhedrin gene promoter of AcNPV, by fusion in the front of intact polyhedrin gene or by insertion into the HindIII site in polyhedrin gene. The recombinant baculoviruses were named as BtrusI or BtrusII, respectively. Although single transcript from the fusion protein gene was apparently observed. BtrusI was produced the two proteins, 92 kDa fusion protein and only polyhedrin. In addition, fusion protein produced by BtrusI did not form polyhedra. Interestingly, however, the cells infected with BtrusII did not show a 33 kDa polyhedrin band as a cells infected with BtrusI. Cells infected with BtrusII were only produced fusion protein, but the polyhedra formed by fusion protein was not observed. To determine the insecticidal toxicity of fusion protein, therefore, Sf9 cells infected with BtrusI were inoculated to Bombyx mori larvae. Sf9 cells infected with BtrusI that expressed the fusion protein caused larval mortality although the insecticidal toxicity was low. In conclusion, our results clearly demonstrated that the fusion protein with polyhedrin and Bt cryIA(c) crystal protein have a insecticida toxicity.

• Journal of Sericultural and Entomological Science
• /
• v.33 no.1
• /
• pp.13-20
• /
• 1991

The flacheric virus(FV) was isolated from the diseased larvae with flaccidity symptoms of the silkworm, Bombyx mori, which were collected in Hyangnam, Kyunggi Province and in Youcheon, Kyungpook Province. The properties of the virus were investigated and compared with the Japanese FV in morphology, size, nucleic acid and structural proteins. The Hyangnam isolate had a diameter of 27$\pm$1, 7nm with speherical shape and contained RNA of which the electrophoretic pattern was same as that of the Japanese FV. The virus had four sturctural proteins and their molecular weight were estimated as 35, 000, 33, 000, 31, 000, and 11, 400 daltons, respectively. The Youcheon isolate had two different sizeds in diameter of 27$\pm$1.7nm and 21$\pm$0.8nm. The antigenicity of the Hyangnam isolate was proved to be identical to that of Japanese FV, whereas the antisera against the Youcheon isolate (mixed with two different sizes) reacted with Japanese FV and densonucleosis virus type 1, respectively. From these characteristic of the isolated viruses, it was concluded that the Hyangnam isolate was identical to the Japanese FV and the Youncheon isolate contained the same viruses as the Japanese FV and DNVI.

• Korean journal of applied entomology
• /
• v.18 no.1 s.38
• /
• pp.1-10
• /
• 1979

An inclusion forming virus isolated from a fan webworm, Hyphantria cunea, in 1975 was identified as a nuclear polyhedrosis virus. With the virus isolated in Korea, it was considered that the virus would be one of the valuable microorganism in microbial control. In this connection, 1) the shape and size of the virus for identification, 2) susceptibility of the various instar larvae to the virus, 3) the effects of storage condition on the pathogenicity and the cross infection of the virus to the larvae of Bombyx mori were examined. The results are summarized as follows; 1. The polyhedron was tetrahedron or hexahedron of $2\mu$ in size and the rod-shaped virus particles consisting of $2\~14$ rods in a bundle were $330m{\mu}\times35m{\mu}$ in size. 2. The hexagonal nuclear polyhedra were found only in the nucleus of the midgut cells but were variable in size. 3. The $LD_{50}$ values for the various instar larvae of H. cunea were $8.377\times10^4\;PIBs/ml$ for the second, $4.974\times10^5\;PIBs/ml$ for the fifth instar larvae. The $LT_{50}values$ for $10^6\;PIBs/ml$ were 9.6 days for the second, 11.5 days for the third, 12.0 days for the fourth and 17 days for the fifth instar larvae. 4. The susceptibility of H. cunea to the nuclear polyhedrosis virus was greater in the first generation than in the second generation. 5. The effect of the storage conditions on the pathogenicity of the nuclear polyhedra was less in refrigerator $(5^{\circ}C)$ and in freezing $(-80^{\circ}C)$ than in room temperature $(18.5^{\circ}C)$, especially as air-dried polyhedra than as suspension. The pathogenicity of the polyhedra seemed to decrease by sunlight during storage as cadavers, since rather greater decrease in pathogenicity was found in sunny condition than in shady condition. 6. The effective spray concentration was $6.4\times10^7\;PIBs/ml$ in the field and its $LT_50$ values for the third and the fifth instar larvae were 4.8 days and 14.2 days, respectively. 7. No cross infections were found in the nuclear polyhedrosis virus between H. cunea and B. mori. larvae.