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bombyx mori bioassay

Manjunatha Ks Gowda, Anitha Peter, Mahantesha Bn Naika, P Niranjana, P Shamprasad
Biotic stress affects all organisms, including the silkworm (Bombyx mori L.) which also has a plethora of defense mechanisms against the infecting pathogens. The midgut plays an important role as a natural barrier and source of innate immunity. The present investigation was thus undertaken to identify an antiviral red fluorescent protein formed in the midgut of the silkworm Bombyx mori L. The digestive juice from fifth instar Kolar Gold silkworm larvae was collected and subjected to 40 % ammonium sulfate precipitation and purified by gel filtration chromatography (GFC)...
January 15, 2018: Protein and Peptide Letters
Zhangfei Shen, Xue Jiang, Lili Yan, Yu Chen, Weiwei Wang, Ying Shi, Liangen Shi, Dongxiang Liu, Naiming Zhou
Diapause hormone (DH) is a 24-aa amidated neuropeptide that elicits the embryonic diapause of the silkworm, Bombyx mori ( Bommo), via sensitive and selective interaction with its receptor, Bommo DH receptor ( Bommo-DHR). Previous studies of the structure-activity relationship of Bommo-DH were all based on an in vivo diapause-induction bioassay, which has provided little information on the structure of Bommo-DHR or its iteration with DH. Here, to unveil the interaction of Bommo-DH with its receptor, N-terminally truncated analogs and alanine-scanning mutants of Bommo-DH were chemically synthesized and functionally evaluated by using a Cy5...
March 2018: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
Weitao Qiu, Jing Wu, Jae-Hoon Choi, Hirofumi Hirai, Hiroshi Nishida, Hirokazu Kawagishi
Two compounds, 3'-deoxyinosine and cordycepin, were isolated from Bombyx mori inoculated with Cordyceps militaris. In the bioassay examining cytotoxicity against cancer cells, both compounds showed toxicity against A549, PANC-1, and MCF-7 cancer cells.
June 2017: Bioscience, Biotechnology, and Biochemistry
R S Sajeevan, Karaba N Nataraja, K S Shivashankara, N Pallavi, D S Gurumurthy, M B Shivanna
Mulberry (Morus species) leaf is the sole food for monophagous silkworms, Bombyx mori L. Abiotic stresses such as drought, salinity, and high temperature, significantly decrease mulberry productivity and post-harvest water loss from leaves influence silkworm growth and cocoon yield. Leaf surface properties regulate direct water loss through the cuticular layer. Leaf surface waxes, contribute for cuticular resistance and protect mesophyll cells from desiccation. In this study we attempted to overexpress AtSHN1, a transcription factor associated with epicuticular wax biosynthesis to increase leaf surface wax load in mulberry...
2017: Frontiers in Plant Science
Rui Yang, Jianjia Zhang, Min Feng, Xiaofeng Wu
Bombyx mori nucleopolyhedrovirus orf58a (bm58a) and its homologues are highly conserved in genomes of all sequenced group I alphabaculoviruses and its function is still unknown. Transcriptional analysis revealed that bm58a is a very late gene initiated from a late transcriptional start motif TAAG. To examine its role in the virus, a bm58a knockout virus (vBmbm-58a-KO-PH-GFP) was generated through homologous recombination in Escherichia coli. Analysis of fluorescence microscopy, titration assays and electron microscopy examination showed that the deletion of bm58a did not affect viral replication and occlusion bodies formation in vitro, indicating that bm58a is not required for viral propagation...
November 2016: Journal of General Virology
Feifei Yin, Ruikun Du, Wenhua Kuang, Guang Yang, Hualin Wang, Fei Deng, Zhihong Hu, Manli Wang
Fibroblast growth factor (FGF) is found throughout multicellular organisms; however, fgf homologs (vfgf) have only been identified among viruses in lepidopteran baculoviruses. The function of vFGFs from Group I alphabaculoviruses, including Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV), involves accelerated killing of infected larvae by both viruses. The vFGF of Group II alphabaculovirus is structurally different from that of Group I alphabaculovirus, with a larger C-terminal region and additional N-linked glycosylation sites...
June 2016: Virologica Sinica
Francesca Berini, Silvia Caccia, Eleonora Franzetti, Terenzio Congiu, Flavia Marinelli, Morena Casartelli, Gianluca Tettamanti
BACKGROUND: The peritrophic matrix (PM) is formed by a network of chitin fibrils associated with proteins, glycoproteins and proteoglycans that lines the insect midgut. It is a physical barrier involved in digestion processes, and protects the midgut epithelium from food abrasion, pathogen infections and toxic materials. Given its fundamental role in insect physiology, the PM represents an excellent target for pest control strategies. Although a number of viral, bacterial and insect chitinolytic enzymes affecting PM integrity have already been tested, exploitation of fungal chitinases has been almost neglected...
May 2016: Pest Management Science
Susumu Katsuma, Toru Shimada
Several lines of evidence have shown that the deletion of the ecdysteroid UDP-glucosyltransferase gene (egt) from the nucleopolyhedrovirus (NPV) genome increases the killing speed of host lepidopteran larvae. However, it has not been investigated in detail whether the effects of egt deletion depend on the larval stages of the host insect. In this study, we performed bioassays using 10 continuous larval stages of the 4th- or 5th-instar Bombyx mori larvae and B. mori NPV egt mutants. The fast-killing phenotype was observed in the egt mutants only when the infection process progressed through larval-larval transition...
March 2015: Journal of Invertebrate Pathology
Wei Sun, Yi-Hong Shen, Min-Jin Han, Yun-Feng Cao, Ze Zhang
Although there are many studies to show a key role of transposable elements (TEs) in adaptive evolution of higher organisms, little is known about the molecular mechanisms. In this study, we found that a partial TE (Taguchi) inserted in the cis-regulatory region of the silkworm ecdysone oxidase (EO) gene, which encodes a crucial enzyme to reduce the titer of molting hormone (20-hydroxyecdysone, 20E). The TE insertion occurred during domestication of silkworm and the frequency of the TE insertion in the domesticated silkworm (Bombyx mori) is high, 54...
December 2014: Molecular Biology and Evolution
Zhan-Qi Dong, Jun Zhang, Xue-Mei Chen, Qian He, Ming-Ya Cao, La Wang, Hai-Qing Li, Wen-Fu Xiao, Cai-Xia Pan, Cheng Lu, Min-Hui Pan
Bombyx mori nucleopolyhedrovirus (BmNPV) ORF79 (Bm79) encodes an occlusion-derived virus (ODV)-specific envelope protein, which is a homologue of the per os infectivity factor 4 (PIF4) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV). To investigate the role of ORF79 in the BmNPV life cycle, a Bm79 knockout virus (vBm(Bm79KO)) was constructed through homologous recombination in Escherichia coli. Viral DNA replication, budded virus (BV) production and polyhedra formation were unaffected by the absence of BM79...
May 12, 2014: Virus Research
Kenichi Ishii, Tatsuo Adachi, Takashi Hara, Hiroshi Hamamoto, Kazuhisa Sekimizu
Injection of culture supernatant of Serratia marcescens, a Gram-negative bacterium pathogenic to a wide range of host animals including insects and mammals, into the hemolymph of silkworm (Bombyx mori) larvae led to continuous flow of the hemolymph (blood of insects) from the injection site. The amount of hemolymph lost within 60 min reached 15-20% of the total larval weight. Using a bioassay with live silkworms, we purified Serralysin, a metalloprotease that requires divalent cations for its activity, as the factor responsible for the promotion of hemolymph bleeding from the culture supernatant of S...
March 2014: Journal of Invertebrate Pathology
Kenichi Ishii, Tatsuo Adachi, Hiroshi Hamamoto, Kazuhisa Sekimizu
Injection of a culture supernatant of Serratia marcescens into the bloodstream of the silkworm Bombyx mori increased the number of freely circulating immunosurveillance cells (hemocytes). Using a bioassay with live silkworms, serralysin metalloprotease was purified from the culture supernatant and identified as the factor responsible for this activity. Serralysin inhibited the in vitro attachment of both silkworm hemocytes and murine peritoneal macrophages. Incubation of silkworm hemocytes or murine macrophages with serralysin resulted in degradation of the cellular immune factor BmSPH-1 or calreticulin, respectively...
February 28, 2014: Journal of Biological Chemistry
Moisés J Zotti, Ellen De Geyter, Luc Swevers, Antônio S K Braz, Luis P B Scott, Pierre Rougé, Josep Coll, Anderson D Grutzmacher, Eder J Lenardão, Guy Smagghe
Ecdysteroid signal transduction is a key process in insect development and therefore an important target for insecticide development. We employed an in vitro cell-based reporter bioassay for the screening of potential ecdysone receptor (EcR) agonistic and antagonistic compounds. Natural ecdysteroids were assayed with ecdysteroid-responsive cell line cultures that were transiently transfected with the reporter plasmid ERE-b.act.luc. We used the dipteran Schneider S2 cells of Drosophila melanogaster and the lepidopteran Bm5 cells of Bombyx mori, representing important pest insects in medicine and agriculture...
November 2013: Pesticide Biochemistry and Physiology
Changlong Shu, Dongming Liu, Zishan Zhou, Jilin Cai, Qi Peng, Jiguo Gao, Fuping Song, Jie Zhang
The cry1-type genes of Bacillus thuringiensis represent the largest cry gene family, which contains 50 distinct holotypes. It is becoming more and more difficult to identify cry1-type genes using current methods because of the increasing number of cry1-type genes. In the present study, an improved PCR-restriction fragment length polymorphism (PCR-RFLP) method which can distinguish 41 holotypes of cry1-type genes was developed. This improved method was used to identify cry1-type genes in 20 B. thuringiensis strains that are toxic to lepidoptera...
November 2013: Applied and Environmental Microbiology
Marta Giner, Jesús Avilla, Mercè Balcells, Silvia Caccia, Guy Smagghe
We investigated the effects of five allyl esters, two aromatic (allyl cinnamate and allyl 2-furoate) and three aliphatic (allyl hexanoate, allyl heptanoate, and allyl octanoate) in established insect cell lines derived from different species and tissues. We studied embryonic cells of the fruit fly Drosophila melanogaster (S2) (Diptera) and the beet armyworm Spodoptera exigua (Se4) (Lepidoptera), fat body cells of the Colorado potato beetle Leptinotarsa decemlineata (CPB) (Coleoptera), ovarian cells of the silkmoth Bombyx mori (Bm5), and midgut cells of the spruce budworm Choristoneura fumiferana (CF203) (Lepidoptera)...
January 2012: Archives of Insect Biochemistry and Physiology
Kevin D Clark, Michael R Strand
The phenoloxidase (PO) cascade regulates the melanization of blood (hemolymph) in insects and other arthropods. Most studies indicate that microbial elicitors activate the PO cascade, which results in processing of the zymogen PPO to PO. PO is then thought to oxidize tyrosine and o-diphenols to quinones, which leads to melanin. However, different lines of investigation raise questions as to whether these views are fully correct. Here we report that hemolymph from the silkmoth, Bombyx mori, rapidly melanizes after collection from a wound site...
May 17, 2013: Journal of Biological Chemistry
Qi Tang, Guohui Li, Qin Yao, Liang Chen, Peng Lv, Chaoqun Lian, Keping Chen
Orf91 (Bm91) of Bombyx mori nucleopolyhedrovirus (BmNPV) is a highly conserved gene that encodes a predicted 105-amino-acid protein, but its function remains unknown. In the current study, 5'-RACE revealed that the transcription initiation site of Bm91 was - 12 nucleotides upstream of the start codon ATG, transcription of Bm91 was detected from 12 to 96 h postinfection (p.i.) and Bm91 protein was detected from 24 to 96 h p.i. in BmNPV-infected BmN cells. Furthermore, Western blot analysis revealed that Bm91 was in occlusion-derived virus (ODV) but not in budded virus (BV)...
May 2013: Journal of Invertebrate Pathology
Katsuhiko Ito, Kurako Kidokoro, Sachiko Shimura, Susumu Katsuma, Keiko Kadono-Okuda
Bombyx mori densovirus type 1 (BmDNV-1) is a pathogen causing flacherie disease in silkworms. BmDNV-1 multiplies only in the nuclei of the columnar cells of larval midgut epithelium. Although several immunohistochemical studies using anti-BmDNV-1 antibody have been reported to date, sequential pathological changes in BmDNV-1-infected larvae have not been completely elucidated. In this paper, sequential investigations were performed on the pathological features of BmDNV-1-infected larvae and BmDNV-1 propagation...
March 2013: Journal of Invertebrate Pathology
Min-Juan Zhang, Ruo-Lin Cheng, Yi-Han Lou, Wan-Lu Ye, Tao Zhang, Xiao-Ying Fan, Hai-Wei Fan, Chuan-Xi Zhang
The Bombyx mori nucleopolyhedrovirus (BmNPV) is a baculovirus that selectively infects domestic silkworm. BmNPV ORF71 (Bm71) is not a core set gene in baculovirus and shares 92 % amino acid sequence identity with Autographa californica multinucleocapsid NPV ORF88 (Ac88/cg30). Previously, it has been reported that virus lacking Ac88 had no striking phenotypes in cell lines or host larvae. However, the exact role of Bm71 during BmNPV life cycle remains unknown. In the present study, we constructed a Bm71-disrupted (Bm71-D) virus and assessed the effect of the Bm71 disruption on viral replication and viral phenotype throughout the viral life cycle...
August 2012: Virus Genes
Susumu Katsuma, Jun Kobayashi, Yasue Koyano, Noriko Matsuda-Imai, WonKyung Kang, Toru Shimada
Lepidopteran nucleopolyhedroviruses (NPVs) show distinct tissue tropism in host insect larvae. However, the molecular mechanism of this tropism is largely unknown. We quantitatively investigated NPV tissue tropism by measuring mRNA levels of viral genes in 16 tissues from Bombyx mori NPV (BmNPV)-infected B. mori larvae and found clear tissue tropism, i.e., BmNPV replicates poorly in the silk glands, midgut, and Malpighian tubule compared with other larval tissues. We next identified the viral genes determining tissue tropism in NPV infection by investigating the phenotypes of larvae infected with 44 BmNPV mutants in which one gene was functionally disrupted by a LacZ cassette insertion...
March 2012: Journal of Virology
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