Quantitative proteomics of the silkworm midgut after bmnpv infected by labelfree method the research material is antiviral nillvr strain. Identification of the proteome of the midgut of silkworm. Analysis of gene expression in the midgut of bombyx mori. The gut lumen is the primary site of digestion and detoxification and thus presents conditions hostile to most proteins.
Molecular and biochemical responses in the midgut of the. To extract acetamiprid in silkworm midgut, 1 g of midgut was grinded, homogenized with 1 ml ethyl acetate and centrifuged at 3000 g for 5 min to collect the supernatant. Comparative proteomic analysis of midgut proteins from. These data will insight into the molecular mechanism of silkworm.
Comparative proteomic analysis of larva and adult heads of. Comparative proteomic analysis reveals the suppressive. Pdf proteomic analysis of the silkworm midgut during. The enzyme activity increased during fifth instar reached a peak, and decreased significantly through prepupal stage to the lowest level at the pupal stagtouring pharate adult period, the protease activity increased to reach another peak just before emergence of the moth. Briefly, rna isolation, amplification, labeling, hybridization, and micro. Comparative subcellular proteomics analysis of susceptible. Investigation of the protein profile of silkworm bombyx mori pupae. Nanoparticulate tio 2 protection of midgut damage in the. Comparative proteomic analysis of midgut proteins from male and.
The mulberry silkworm, bombyx mori lepidoptera, harbors several bacteria in its midgut aiding the metabolic processes. Impacts on silkworm larvae midgut proteomics by transgenic. One of the tubes situated on the dorsal side bra are three pairs of m alpighian tubes in the silkworm larva that stick to the midgut and run towards the. Comparative subcellular proteomics analysis of susceptible and nearisogenic resistant bombyx mori lepidoptera larval midgut response to bmnpv infection. Shangzhi zhang, jiaping xu and erjun ling, metabolic changes in the midgut of eri silkworm after oral administration of 1deoxynojirimycin. Unlimited viewing of the articlechapter pdf and any associated supplements and figures.
Pdf the midgut is the major organ for food digestion, nutrient absorption and also a barrier for foreign substance. The 5thinstar larval stage of silkworm is very important for larval growth, development, and silk production. Impacts on silkworm larvae midgut proteomics by transgenic trichoderma strain and analysis. A luteinbinding protein was purified from fifth instar larval midgut of bombyx mori by a combination of ammonium sulfate fractionation and three chromatographic procedures, gel filtration, chromatofocusing, and anion exchange chromatography. Proteomic analysis of larval integument, trachea and adult. Read comparative proteomic analysis reveals the suppressive effects of dietary high glucose on the midgut growth of silkworm, journal of proteomics on deepdyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips. Beerntsen2, peng zhang3, chengshu wang1, erjun ling1 1key laboratory of insect developmental and evolutionary biology, institute of plant physiology and ecology, shanghai. A total of 793 deps were identified after comparing the four proteomes.
Mapping the larval midgut lumen proteome of helicoverpa. A 1hnmrbased metabonomic study, plos one, 12, 3, e01732, 2017. We also found that the silkworm larval midgut responded to starvation by producing a 10 kda heat shock protein and a diapause hormone precursor. To understand the molecular mechanism, comparative proteomic approach was employed to. The protease activity in the midgut of bivoltine and muitivoltine races of the silkworm,bombyx mori, was studied. A total of 90 proteins were identified and 79 proteins were described for the first time. Proteomic analysis of silkworm midgut cellular proteins. Proteome analysis of silk gland proteins from the silkworm, bombyx mori. The brush border membrane bbm aminopeptidase n was isolated from tritonsolubilized bbm by column chromatography at a specific activity of 61. Proteomic analysis of larval midgut from the silkworm bombyx mori by ningjia he, zhonghuai xiang, ling jia, qiang fu, yunmin xu and sai zhang no static citation data no static citation data cite. On the first day of fifth instar, all larvae were starved for 24 h. In addition, gene expression profile in the head of silkworm at different. Proteomics analysis of digestive juice from silkworm.
Collectively, the comparative proteomics analyses indicate that mulberry leaves not only provide amino acids to the silkworm but also display defense against silkworm feeding, although the silkworm grows very well by feeding on mulberry leaves, which provides new insights into the interactions between hostplant and insect herbivores. Proteomics analysis of fat body from silkworm bombyx mori. Among the 85 differentially expressed proteins identified, six were in the bmnpv group and six were in the control group. The midgut is formed by a monolayered epithelium supported by a basal lamina. Proteomic analysis of the silkworm midgut during larval. In order to understand the biological function of silkworm larval pm, shotgun liquid chromatography tandem mass spectrometry. Comprehensive profiling of lysine acetylome in baculovirus. Comparative subcellular proteomics analysis of susceptible and. This was the first report using an itraq approach to analyze proteomes of the silkworm midgut against bmcpv infection, which contributes to understanding the defense mechanisms of silkworm midgut to virus infection.
Proteomicsbased approaches have been recognized as powerful tools to study proteomic changes. Proteomic analysis of larval midgut from the silkworm. In order to compare those data directly, transcription changes of the molting midgut were analyzed using the same method. Comprehensive profiling of lysine acetylome in baculovirus infected silkworm bombyx mori cells dongbing hu. The larval midgut of the silkworm represents the middle part of the alimentary canal and is responsible for the digestion and the assimilation of nutrients that are necessary for the growth and the development of the larva dow, 1986. Responses of detoxification enzymes in the midgut of. Bombyx mori cytoplasmic polyhedrosis virus bmcpv specifically infects the epithelial cells in the midgut of silkworm and causes them to death, which negatively affects the sericulture industry. Proteomic analysis of silkworm hemolymph at the fifthinstar day3 larva stage was performed.
Proteomic analysis of larval midgut from the silkworm bombyx mori. In this study, to investigate the mechanism underlying variation of nutrient utilization in the midgut of silkworm and difference in growth rate between male and female silkworms, we used comparative proteomic approach to detect differentially expressed midgut proteins between two sexes on the second day of fifth instar. In order to obtain an overall view of silkworm response to bmcpv infection, we performed a proteomic analysis of the midgut of silkworm responses to bmcpv infection by itraq. Tandem searching software using different parameters and validated by using the poisson model. Nanoparticulate np titanium dioxide nanotio 2 has been shown to attenuate damages in b. The 5thinstar larval stage of silkworm is very important for larval growth. In silkworm special excretory organs called m malpighian tubes arise at the junction of small intestine and colon in the hind the tube arising on each side of the hind branches arise. However, the role of proteins and factors involved in. On treating silkworm larvae with pesticides insecticide and herbicide, expression of the bmgsts2 gene increased noticeably in the midgut and reached a peak at 6 to 12 h. The protein expression profile of midgut from the fifth instar of the silkworm was constructed by 2de and showed that over 600 spots were obtained, most of which were distributed in the area from. Proteomics provides insight into the interaction between.
Molecular and biochemical responses in the midgut of the silkworm, bombyx mori, infected with nosema bombycis. Alkaline protease in the midgut of the silkworm bombyx. Purification and partial characterization of a lutein. Labelfree proteomic analysis of silkworm midgut infected. Differential proteomics of the larval midgut after bmnpv infection. However, little is known about the molecular mechanisms of midgut injury due to organophosphorus insecticide exposure and. Comparative proteomic analysis between the domesticated. To gain an insight into the effects of different diets on growth and development of the domesticated silkworm at protein level, we employed comparative proteomic approach to investigate the proteomic differences of midgut, hemolymph, fat body and posterior silk gland of the silkworms reared on fresh mulberry leaves and on artificial diet. Tandem searching software using different parameters and. We utilized the proteomic approach to investigate the proteome of the fifth instar hemolymph during. Quantitative proteomics of the silkworm midgut after bmnpv infection by itraq. Proteome analysis of silk gland proteins from the silkworm. The properties of aminopeptidase n in bombyx mori larval midgut are described.
Several papers concerned with gene regulation of silkworm midgut development and immunity have been published using microarray assays 18, 20, 21. The midgut is the digestive apparatus of the silkworm and its proteome was studied by using nanolc liquid chromatography electrospray ionization msms tandem ms. Previous studies have analyzed the midgut transcriptome and proteome after challenge with bombyx mori nucleopolyhedrovirus bmnpv, however little information is available on the digestive juice proteome after bmnpv challenge. Read proteomic analysis of silkworm midgut cellular proteins interacting with the 5. A total of 162, 259, and 175 peptides from silkworm larval integument and trachea, and adult scale obtained by database search were matched to 48, 51, and 40 proteins, respectively. Pdf proteomic analysis of larval midgut from the silkworm. Therefore, research on the silkworm midgut proteome is essential for developing new lepidopteran pest control strategies.
One tube in one group runs along the dorsal and turns backwards at the 4th abdominal segment and second tube runs along midgut to laterally and turns backwards at the point of center of 4. Owing to the increasing interest in insects as a new food and feed protein. A total of 4194 host proteins are quantified, of which 33 are upregulated and 47 are. The insect peritrophic membrane pm, separating midgut epithelium and intestinal contents, is protective lining for the epithelium and plays the important role in absorption of nutrients, and also is the first barrier to the pathogens ingested through oral feeding. It is an entry for pathogens, toxins, and pesticides. The result showed that there had been target band in silkworm midgut, fat body, silk gland, and hemolymph of normal diet group for idgf, whereas no target band in the midgut of highglucose diet silkworms.
Some key deps were validated by rtqpcr and western blot. The midgut is the major organ for food digestion, nutrient absorption and also a barrier for foreign substance. This suggested idgf expression is inhibited in midgut by highglucose diet, which was consistent with 2de result. Over the past few years, researchers have used suppression subtractive hybridization ssh, dna microarray system, high throughput expressed sequence tags, and isobaric tag for relative and absolute quantization itraq proteomics based methods to examine changes in gene transcription in the midgut of the silkworm b. Transcriptional profiling of midgut immunity response and degeneration in the wandering silkworm, bombyx mori qiuyun xu1. Pdf properties of the aminopeptidase n from the silkworm.
The 5thinstar larval stage of silkworm is very important for. Bacillus thuringiensis bt is a unique bacterium in that it shares a common place with a number of chemical compounds which are used commercially to control insects important to agriculture and public health. This was the first report using an itraq approach to analyze proteomes of the silkworm midgut against bmcpv infection, which contributes to understanding the defense mechanisms of silkworm midgut to virus. In order to determine the midgut response at the protein levels to the virus infection, differential proteomes of the silkworm midgut responsive to bmcpv infection were identified with isobaric tags for relative and. Silkworm midgut cellular proteins were identified that associate with an rna containing sequences from the 5.
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