Resistance to Bombyx mori Densonucleosis Virus Type 1 and Its Inheritance in Silkworm, Bombyx mori L.

Sen, Ratna,Nataraju, B.,Balavenkatasubbaiah, M.,Premalatha, V.,Thiagarajan, V.,Datta, R.K. Korean Society of Sericultural Science 2004 International Journal of Industrial Entomology Vol.9 No.1

Bombyx mori densonucleosis virus type 1 (BmDNV1)- a non occluded virus causes flacherie disease in the susceptible stocks of the silkworm, Bombyx mori. However, some stocks are non-susceptible. Non-susceptibility to BmDNV1 in B. mori is a unique case where the virus infection is completely inhibited by a single gene of the host. A survey conducted by this institute in some parts of Karnataka state has revealed that, 43.05% of the total incidence of flacherie disease caused by non-occluded viruses, are due to the synergistic infection of B. mori densonucleosis and infectious flacherie virus. Earlier study indicated that rearing of BmDNV1 resistant silkworm stock is effective in protecting silkworm against BmIFV also. In the present study the response of 78 silkworm stocks which include 42 of non-diapausing and 36 of diapausing groups, to BmDNV1 is investigated. Newly ecdysed third instar larvae were inoculated per-os with 10% inoculum of BmDNV1 extracted from the mid-gut of infected silkworm. One non-diapausing and three diapausing silkworm stocks were found to be resistant to BmDNV1. Eleven silkworm stocks were found to possess moderate resistance whereas rest sixty three were found to be susceptible to BmDNV1. Genetic analysis has shown that the resistance to BmDNV1 is autosomally inherited and controlled by a major dominant or a major recessive gene in different silkworm stocks. These resistant stocks can be utilized as the resource material to develop BmDNV1 resistant commercial hybrids. The selection strategies, depending upon the mode of inheritance of resistance in the resource material chosen, are discussed.

집누에(Bombyx mori)와 멧누에(Bombyx mandayina)의 種間交雜에 있어서 卵殼構造 및 Chorion 蛋白質

金鍾吉,盧時甲 한국잠사학회 1994 한국잠사곤충학회지 Vol.36 No.1

Chorion(egg-shell) morphology of the F1 hybrid between Bombyx mori and Bombyx mandarina has been observed by scanning electron microscope and chorion protein was analyzed by electrophoresis. The chorion surface structure of F1, hybrids in the lateral (flat) region was similar to that of maternal line. The F1, hybrids chorion was found to have basically a three layer structure. The middle and inner layer were very much like those of the Bombyx mandarina and Bombyx mori. There were many conic pillar structures in the outer layer of the F1, hybrid, which was similar to Bombyx mandarina. This conic pillar structure had a thin cover layer was more clear in the dorsal and ventral side of the F1, hybrid chorion. The conic pillar structure of Bombyx mandarina was found to be dominant in F1 hybrid chorion irrespective of their maternal line. Major components of chorion protein were analyzed by two-dimensional electrophoresis and found to have isoelectric points in the range of pH 4.0-6.5 and molecular weight 10 to 50 kd. F1 hybrid chorion protein components related directly to those of the maternal line. The conic pillar structure was dominat characteristic and it was present in all F1 hybrid.

Role of Toll-receptors in the inhibition of BmNPV proliferation and their interplay with antimicrobial seroin proteins in Bombyx mori

C.P. Singh,J. Singh,K.P. Arunkuma 한국응용곤충학회 2019 Journal of Asia-Pacific Entomology Vol.22 No.3

The domesticated silkworm, Bombyx mori, is an economically important insect that majorly contributes in financially viable sericulture industry. The baculovirus, Bombyx mori nuclear polyhedrosis virus (BmNPV) is a deadly virus that infects B. mori and inflicts heavy loss to silk production by causing high mortality of silkworms. A number of studies have been done to identify antiviral genes and pathways against BmNPV infection in B. mori. Toll-pathway is a well-known immune pathway responsible for combating bacterial and fungal infections in silkworm. However, the role of Toll-receptors in inhibiting BmNPV has not been explored. In the present study, we have attempted to examine the role of Toll-receptors in inhibiting BmNPV proliferation in B. mori larvae. The effect of Toll-receptors on viral proliferation was determined by quantifying viral DNA and counting viral occlusion bodies upon RNA interference-mediated knockdown of Toll-receptors in B. mori larvae. Knockdown of BmToll, BmToll-6, BmToll-7, BmToll-8 and BmToll-9 led to higher accumulation of BmNPV. Further, interplay between Toll-receptors and the antimicrobial proteins of B. mori, called seroins were studied using RNA interference. As a result, we found a downregulation of Seroin1 transcript level upon knockdown of BmToll, BmToll-7, BmToll-8, BmToll-10, BmToll-11 and BmToLK2, whereas Seroin2 expression was found to decrease only upon BmToll-7 knockdown. Moreover, knockdown of seroins did not result in clearly interpretable expression patterns of Toll-receptors. These results suggest the involvement of B. mori Toll-pathway in defense against BmNPV infection and provides a new insights into the antiviral defense mechanism of B. mori.

Sensory Profiles of Protein-Fortified Kefir prepared Using Edible Insects (Silkworm Pupae, Bombyx mori) : A Preliminary Study

( Young-ji Kim ),( Jung-whan Chon ),( Kwang-young Song ),( Dong-hyeon Kim ),( Hyunsook Kim ),( Kun-ho Seo ) 한국유가공기술과학회 2017 Journal of Dairy Science and Biotechnology (JMSB) Vol.35 No.4

Bombyx mori (silkworm pupae) is a unique and biologically significant insect, which is a recognized source of high quality protein that provides all the essential amino acids required for human health. Recently, many studies have focused on various biomedical applications of B. mori proteins. The purpose of this study was to manufacture protein-fortified kefir containing different concentrations of B. mori powder according to pH and sensory evalua-tions. The value of the protein-fortified kefir increased but the pH decreased with increasing incubation time, indicating that the amount of B. mori powder did not affect and pH. Addition of B. mori powder also did not affect the sensory properties of overall acceptability, texture, and color compared to control group without addition of B. mori powder. However, flavor and taste were affected by increasing the amounts of B. mori powder, with a significant difference in both flavor and taste between the control and treated groups (both p<0.05). There was no significant difference in overall acceptability, texture, and color. Fur-ther studies are needed for producing kefir as a dietary supplement utilizing the functional properties of B. mori.

COI 염기서열 기반 백강잠 신속 감별용 SCAR marker 개발 - 백강잠 유전자 감별 -

김욱진,양선규,노푸름,박인규,최고야,송준호,문병철 대한본초학회 2019 大韓本草學會誌 Vol.34 No.5

Objectives : To ensure the safety, quality and pharmacological efficacy of Batryticatus Bombyx, it is important to discriminate with adulterants. In Korean Herbal Pharmacopoeias (KHP), the authentic species of Batryticatus Bombyx is defined only Bombyx mori. Therefore, the aim of this study is establishment of PCR assay method using the sequence characterized amplified region (SCAR) marker based on COI DNA barcode for discriminating six species related to Batryticatus Bombyx. Methods : Seventeen samples of six species (Bombyx mori, Bombyx mandarina, Rhodinia fugax, Oberthueria caeca, Actias artemis, and Caligula japponica) were collected from different habitate and nucleotide sequences of cytochrome c oxidase subunit I(COI) barcode regions were analyzed by Sanger sequencing methods. To develop SCAR-based PCR assay method, we designed species-specific primers based on COI sequence variabilities and verified those specificities using 17 samples of six species as well as commercial herbal medicines. Results : In comparative multiple analysis of COI sequences, six species were distinguished by species-specific nucleotides at the species level. To develop rapid and reliable PCR assay method for genetic authentication of Batryticatus Bombyx, therefore, we designed species-specific SCAR primers based on these nucleotide sequences and confirmed those specificities. Using these SCAR primers, We also established simple conventional PCR assay method using these SCAR primers at the species level. Conclusions : The comparative analysis of COI sequences and SCAR-based PCR assay methods represented equal results for distinguishing authentic Batryticatus Bombyx and adulterations at the species level. Therefore, our results are expected protecting adulteration of herbal medicine Batryticatus Bombyx.

Sensory Profiles of Protein-Fortified Kefir prepared Using Edible Insects (Silkworm Pupae, Bombyx mori) : A Preliminary Study

김영지,전정환,송광영,김동현,김현숙,서건호 한국낙농식품응용생물학회 2017 한국유가공기술과학회지 Vol.35 No.4

Bombyx mori (silkworm pupae) is a unique and biologically significant insect, which is a recognized source of high quality protein that provides all the essential amino acids required for human health. Recently, many studies have focused on various biomedical applications of B. mori proteins. The purpose of this study was to manufacture protein-fortified kefir containing different concentrations of B. mori powder according to pH and sensory evaluations. The value of the protein-fortified kefir increased but the pH decreased with increasing incubation time, indicating that the amount of B. mori powder did not affect and pH. Addition of B. mori powder also did not affect the sensory properties of overall acceptability, texture, and color compared to control group without addition of B. mori powder. However, flavor and taste were affected by increasing the amounts of B. mori powder, with a significant difference in both flavor and taste between the control and treated groups (both p<0.05). There was no significant difference in overall acceptability, texture, and color. Further studies are needed for producing kefir as a dietary supplement utilizing the functional properties of B. mori.

Sensory Profiles of Protein-Fortified Kefir prepared Using Edible Insects (Silkworm Pupae, Bombyx mori) : A Preliminary Study

Kim, Young-Ji,Chon, Jung-Whan,Song, Kwang-Young,Kim, Dong-Hyeon,Kim, Hyunsook,Seo, Kun-Ho Korean Society of Dairy Science and Biotechnology 2017 Journal of Dairy Science and Biotechnology (JMSB) Vol.35 No.4

Bombyx mori (silkworm pupae) is a unique and biologically significant insect, which is a recognized source of high quality protein that provides all the essential amino acids required for human health. Recently, many studies have focused on various biomedical applications of B. mori proteins. The purpose of this study was to manufacture protein-fortified kefir containing different concentrations of B. mori powder according to pH and sensory evaluations. The value of the protein-fortified kefir increased but the pH decreased with increasing incubation time, indicating that the amount of B. mori powder did not affect and pH. Addition of B. mori powder also did not affect the sensory properties of overall acceptability, texture, and color compared to control group without addition of B. mori powder. However, flavor and taste were affected by increasing the amounts of B. mori powder, with a significant difference in both flavor and taste between the control and treated groups (both p<0.05). There was no significant difference in overall acceptability, texture, and color. Further studies are needed for producing kefir as a dietary supplement utilizing the functional properties of B. mori.

Study of gut bacterial diversity of Bombyx mandarina and Bombyx mori through 16S rRNA gene sequencing

Dhiraj Kumar,Zhenli Sun,Guangli Cao,Renyu Xue,Xiaolong Hu,Chengliang Gong 한국응용곤충학회 2019 Journal of Asia-Pacific Entomology Vol.22 No.2

The wild silkworm B. mandarina is living in the natural environment has a strong stress resistance and adaptability after harsh natural selection. The indoor rearing or domestication of the wild silkworm under artificial custody for long period deteriorates stress resistance and ecological adaptability. Therefore, we aimed to investigate the effects of artificial domestication and evolutionary pressure on the gut bacterial diversity of B. mandarina and B. mori. The intestinal content of 6th day of fifth instar B. mandarina and B. mori larvae were analyzed by sequencing of the 16S rRNA gene through Illumina miseq sequencing technology. The outcome of the study revealed that abundance of predominant bacteria of phylum Firmicutes were respectively 81.40% and 81.85% in the late fifth instar silkworm larvae (6th day) of B. mandarina and B. mori. In Firmicutes, abundance of predominant bacterial genus Enterococcus in B. mandarina (69.73%) was comparatively higher than B. mori (48.99%). The genus Advenella belongs to phylum Proteobacteria was recorded only in B. mandarina (11.54%). The abundance of Unclassified_Peptostreptococcaceae, Methanobrevibacter, Ignatzschineria, Petrimonas and Proteiniphilum in B. mandarina were between 0.12 and 0.17%, nevertheless, these bacterial genera were not detected in B. mori. The abundance of genera Lactococcus, Bacillus and Pseudomonas in B. mori (17.73%, 5.02%, and 1.61%) were remarkably higher than B. mandarina (0.15%, 0.54% and 0.45%). These results indicated that substantial difference was observed between the intestinal bacteria of B. mori and B. mandarina population, and structure of the intestinal bacteria could be affected by the artificial domestication and evolutionary pressure.

Bombyx mori b-tubulin Promoter for High-level Expression of Heterologous Genes

( Kwanho Park ),( Tae-won Goo ) 한국잠사학회 2019 International Journal of Industrial Entomology Vol.39 No.1

We previously isolated 9 clones that show stronger signal compared to Bombyx mori cytoplasmic actin gene (BmA3) by using a dot blot hybridization. In this study, we focused on one clone among these clones which has high amino acid similarity with b-tubulin gene of B. mori. This clone was ubiquitously expressed in all tissues and developmental stage of B. mori. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-750/-1) in the 5’-flanking region of b-tubulin gene, which has about 47 fold more intensive promoter activity than BmA3 promoter. Moreover, the b-tubulin promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that b-tubulin promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter. © 2019 The Korean Society of Sericultural Sciences Int. J. Indust. Entomol. 39(1), 22-28 (2019)

Expression analysis of decapentaplegic gene in the silkworm, Bombyx mori

김성렬,최광호,김기영,구태원,김익수,박승원 한국곤충학회 2016 Entomological Research Vol.46 No.6

The decapentaplegic (dpp) gene in Drosophila is involved in multiple developmental processes, and is a highly conserved among various eukaryotic species, including Bombyx mori. Although the gene has well been characterized in Drosophila species the B. mori dpp has not yet been functionally analyzed. In this study, we analyzed the expression pattern of B. mori dpp in 12 different developmental days/stages (7 days for fifth instar larvae, 2 days for spinning stage, 2 days for pupal stages, and 1 day for adults) in both male and female silkworms using quantitative real‐time RT‐PCR (qRT‐PCR). mRNA expression of B. mori dpp was much higher in the female larvae up to the mid‐stage of the fifth instar compared with the corresponding male larvae. Similarly, dpp expression also was much higher in females during the eclosion period than that in the corresponding male pupae. During the embryonic stage, the expression level of the dpp gene was much higher compared to that of adult stage in both male and female silkworms. These results suggest that the B. mori dpp gene plays multiple roles in the developmental of B. mori.