• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.2
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• pp.221-229
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• 2003

Ten multivoltine female parental lines were crossed with popular bivoltine male silkworm breed NB$_4$D$_2$. Three types of heterosis parameters viz., heterosis over mid-parental value (hybrid vigour), heterobeltiosis (useful heterosis) and standard heterosis (standard check) were estimated for 15 economically important quantitative traits. The interaction among the hybrids and parents indicated significant effect for maximum characters. The heterotic effect of new hybrid combination was compared with popular hybrids viz., Pure Mysore${\times}$NB$_4$D$_2$ and Nistari${\times}$NB$_4$D$_2$. Varied heterotic effect was observed for different traits for different hybrid combination. The results inferred that the crosses viz., BL$_{23}$${\times}$NB$_4$D$_2$ ranked top for 14 traits followed by Hosa Mysore${\times}$NB$_4$D$_2$ for 11 traits; PA$_{12}$${\times}$NB$_4$D$_2$ for 9 traits; BL$_{24}$${\times}$NB$_4$D$_2$ for 8 traits; Kolar Gold${\times}$NB$_4$D$_2$ for 7 traits; WAI$_1$${\times}$NB$_4$D$_2$ for 6 traits and MU$_{11}$${\times}$NB$_4$D$_2$ for 5 traits. Among these, the best hybrids Kolar Gold${\times}$NB$_4$D$_2$ and MU$_{11}$${\times}$NB$_4$D$_2$ were identified for longer filament length and fine denier. Similarly for higher cocoon yield and silk productivity BL$_{223}$${\times}$NB$_4$D$_2$ and BL$_{24}$${\times}$NB$_4$D$_2$ were found to be superior. These hybrid combinations are suitable for commercial exploitation at large scale.e.e.e.

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.2
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• pp.123-127
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• 2001

In order to introduce bivoltine races in a tropical country like India, it is necessary to have stability in cocoon crop under high temperature environments. Unlike any temperate country like Japan, the rearing conditions/environment, climatic conditions, quality of mulberry leaf and incidence of diseases are unpredictable in India. Geneticists and breeders of all the sericultural countries have experienced the influence of environment during the process of breeding. in order to select efficiently the breeds with high temperature tolerance, it is verb important to analyse clearly the heritability nature of high temperature tolerance. In light of the above, the present study was undertaken to determine the effect of high temperature treatment of (A) $35{\pm}1^{\circ}C$ and 85${\pm}$5% RH for 24 hrs continuously, (B) $35{\pm}1^{\circ}C$ and 85${\pm}$5% RH for 48 hrs continuously and (C) the control (25${\pm}1^{\circ}$ and 65${\pm}$5% RH in the normal rearing condition from the $3^{rd}of5^{th}$ instar on the pure races such as Moria, Nl37 and Cl46 as well as their Fl hybrids. The overall performance indicate that the hybrids are mare tolerant than the pure races. it was also observed that the overall performance declined in those batches where 48 hrs treatment was given. The most interesting observation noticed in this study was that there was maternal effect regarding temperature tolerance as evident from the better performance of those hybrids where the female parent used was more tolerant as pure race.

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.2
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• pp.107-110
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• 2001

Variations in protein and nucleic acid concentrations were observed in 24 hrs old eggs and hatched larvae of Nistari strain, Bombyx mori, exposed to starvation. Three starvation treatments of 24,48 and 60 hrs were given separately from 0 hr old fifth instar larvae. Biochemical variations were studied in the resultant hatched larvae of one time starved parent, while the eggs obtained from parents receiving starvation in two successive generations were considered for the study. In hatched larvae, protein levers in 24 hrs starvation groups remained significantly higher over control (never starved) while the same was found to be lower in 48 and 60 hrs starvation individuals. The RNA concentration remained significantly higher in all the treated lots. However, DNA content was not found to be significantly altered in hatched larvae after exposure to feeding stress. Protein, RNA and DNA concentration of 24 hrs old eggs produced by all the starved groups of Nistari, which had deceived two consecutive starvation during parental generations, showed higher concentrations of these biomolecules over control. Hence, starvation induced alterations in protein and nucleic acids in eggs and hatched Iarvae are indicative of a preparatory phase adopted by the insect to acclimatise itself and its progeny to stress situations.

• Journal of Sericultural and Entomological Science
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• v.40 no.2
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• pp.111-116
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• 1998

To construct transformed Bm5 cells, Autographa californica nuclear polyhedrosis virus (AcNPV)IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% uncleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/$m\ell$ G418 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistnt cells was isolated and characterized by PCR using AcNPV IE1 gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. moridrived Bm5 cells as well as Spodaptera frugiperda-derived Sf9 cells to produce stably-transformed insect cells.

• Journal of Sericultural and Entomological Science
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• v.46 no.1
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• pp.28-31
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• 2004

Bombyx mori silk fibroin/hyaluronic acid blend films were prepared by mixing aqueous solution of B. mori silk fibroin and hyaluronic acid. According to the FTIR spectra, no interaction between silk fibroin and hyaluronic acid was found. The conformation of silk fibroin in blend films was changed from random coil to $\beta$-sheet structure by treatment of EDC ethanol solution. Thermal degradation peak of silk fibroin and hyaluronic acid was also not altered by blending each other. The average swelling ratio of silk fibroin/hyaluronic acid blend films was 70. Therefore, silk fibroin/hyaluronic acid blend films might be one of possible wound dressing materials.

• International Journal of Industrial Entomology and Biomaterials
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• v.27 no.2
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• pp.282-288
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• 2013

In nature, the population of Nosema bombycis (Microsporidia) causing pebrine disease is small and their development is extremely slow and only few ultimately producing spores. Pebrine infected silkworm, Bombyx mori larvae collected from sericulture field were alive till $3^{rd}$ generation though the concentration of N.bombycis spore was very high ($2.4-3.0{\times}10^8$ spores. $mL^{-1}$). All larvae were died during $4^{th}$ generation with extremely high concentration of pebrine spores ($3.0-4.0{\times}10^9$ spores. $mL^{-1}$) and mostly contain long polar tube (LT). Alternately, all larvae were died immediately (at $3^{rd}$ stage of $1^{st}$ generation) when it was artificially inoculated with same concentration of N.bombycis spores harvested from field ($2.4-3.0{\times}10^8$ spores. $mL^{-1}$) though concentration of spores harvest was very less ($3.0-4.0{\times}10^6$ spores. $mL^{-1}$) and mostly contain short polar tube (ST). Artificially pebrine infected male moth when mated with healthy female moth took six generations to develop pebrine disease and all larvae were died at the $2^{nd}$ stage with very less spore harvest ($3.0-10.0{\times}10^6$ spores. $mL^{-1}$). Survival percentage was increased in all generations (~92.0% at $4^{th}$ generation) when silkworm rearing was conducted under new integrated disease management system.

• Journal of Life Science
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• v.13 no.5
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• pp.596-603
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• 2003

Cells respond to an accumulation of unfolded proteins in the endoplasmic reticulum (ER) by increasing transcription of genes encoding molecular chaperones and folding enzymes. The information is transmitted from the ER lumen to the nucleus by intracellular signaling pathway, called the unfolded protein response (UPR). To obtain genes related to UPR from B. mori, the cDNA library was constructed with mRNA isolated from Bm5 cell lines in which N-glycosylation was inhibited by tunicamycin treatment. From the cDNA library, we selected 40 clones that differentially expressed when cells were treated with tunicamycin. Among these clones, we have isolated ATFC gene showing similarity with Hac1p, encoding a bZIP transcription factor of 5. cerevisiae. Basic-leucine zipper (bZIP) domain in amino acid sequences of ATFC shared homology with yeast Hac1p. Also, ATFC is up-regulated by accumulation of unfolded proteins in the ER through the treatment of ER stress drugs. Therefore we suggest that ATFC represents a major component of the putative transcription factor responsible for the UPR leading to the induction of ER-localized stress proteins.

• Journal of Life Science
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• v.29 no.5
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• pp.537-544
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• 2019

Genome editing technology employing gene scissors has generated interest in molecular breeding in various fields, and the development of the third-generation gene scissors of the clustered, regularly interspaced short palindromic repeat (CRISPR) system has accelerated the field of molecular breeding through genome editing. In this study, we analyzed the possibility of silkworm molecular breeding using gene scissors by genomic and phenotypic analysis after editing the biogenesis of lysosome-related organelles (BmBLOS) gene of Bakokjam using the CRISPR/Cas9 system. Three types of guide RNAs (gRNA) were synthesized based on the BmBLOS gene sequence of Bakokjam. Complexes of the prepared gRNA and Cas9 protein were formed and introduced into Bombyx mori BM-N cells by electroporation. Analysis of the gene editing efficiency by T7 endonuclease I analysis revealed that the B4N gRNA showed the best efficiency. The silkworm genome was edited by microinjecting the Cas9/B4N gRNA complex into silkworm early embryos and raising the silkworms after hatching. The hatching rate was as low as 18%, but the incidence of mutation was over 40%. In addition, phenotypic changes were observed in about 70% of the G0 generation silkworms. Sequence analysis showed that the BmBLOS gene appeared to be a heterozygote carrying the wild-type and mutation in most individuals, and the genotype of the BmBLOS gene was also different in all individuals. These results suggest that although the possibility of silkworm molecular breeding using the CRISPR/Cas9 system would be very high, continued research on breeding and screening methods will be necessary to improve gene editing efficiency and to obtain homozygotes.

• Journal of Microbiology and Biotechnology
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• v.25 no.3
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• pp.386-392
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• 2015

A novel recombinant bacmid, bEasyBm, that enables the easy and fast generation of pure recombinant baculovirus without any purification step was constructed. In bEasyBm, attR recombination sites were introduced to facilitate the generation of a recombinant viral genome by in vitro transposition. Moreover, the extracellular RNase gene from Bacillus amyloliquefaciens, barnase, was expressed under the control of the Cotesia plutellae bracovirus early promoter to negatively select against the nonrecombinant background. The bEasyBm bacmid could only replicate in host insect cells when the barnase gene was replaced with the gene of interest by in vitro transposition. When bEasyBm was transposed with pDualBac-EGFP, the resulting recombinant virus, EasyBm-EGFP, showed high levels of EGFP expression efficiency compared with that of non-purified recombinant virus BmGOZA-EGFP, which was constructed using the bBmGOZA system. In addition, nonrecombinant backgrounds were not detected in unpurified EasyBm-EGFP stocks. Based on these results, a high-throughput system for the generation of multiple recombinant viruses at a time was established.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6351-6356
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• 2013

Background: Goshajinkigan (GJG) is used for the treatment of several neurological symptoms. We investigated the efficacy of GJG and mecobalamin (B12) against neurotoxicity associated with docetaxel (DOC) in breast cancer patients. Materials and Methods: Sixty breast cancer patients were treated with DOC. Thirty-three patients (GJG group) received oral administration of 7.5 g/day GJG and 27 patients (B12 group) received oral administration of 1500 ${\mu}g/day$ B12. Neuropathy was evaluated according to DEB-NTC (Neurotoxicity Criteria of Debiopharm), Common Terminology Criteria for Adverse Events (NCI-CTC) ver. 3.0, and a visual analogue scale (VAS). This study employed a randomized open design. Results: The incidence of neuropathy was 39.3% in the GJG group, and 88.9% in the B12 group (p<0.01). In the GJG group, grade 1 DEB-NTC was observed in 2 cases, grade 2 in 5 cases and grade 3 in 5 cases. Grade 1 NCI-CTC was observed in 7 cases, grade 2 in 6 cases, and VAS was $2.7{\pm}2.2$. In the B12 group, grades 1, 2 and 3 DEB-NTC were observed in one case, 12 cases and 12 cases, respectively; and grades 1, 2 and 3 NCI-CTC were observed in 11 cases, 12 cases and one case, and VAS was $4.9{\pm}2.4$. Conclusions: Concomitant administration of GJG is useful in preventing neuropathy in breast cancer patients treated with a DOC regimen.