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J Invertebr Pathol ; 170: 107323, 2020 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-31926972


Bombyx mori nucleopolyhedrosis virus (BmNPV) has always been a great challenge to the development and stability of the sericulture industry. LncRNAs have been reported to play important roles in gene expression regulation, development and immune response but the roles of lncRNAs in BmNPV infection and silkworm-BmNPV interaction are not clear. We used a genome-wide transcriptome analysis to identify the lncRNAs in Bombyx mori cells (BmN cells) and analyzed the differentially expressed lncRNAs, microRNAs and protein-coding genes in silkworm cells with or without BmNPV infection. A total of 13,159 candidate lncRNAs were identified in the BmN cells, among which 4450 lncRNAs were differentially expressed (DE) with 2837 up-regulated and 1613 down-regulated. In addition, 66 differentially expressed miRNAs (DEmiRNAs) and 7448 differentially expressed mRNAs (DEmRNAs) were identified, and DElncRNA-DEmiRNA-DEmRNA regulatory network was constructed. Gene expression was variable in 4973 of predicted lncRNA cis target genes in BmNPV infected cells. KEGG pathway analysis indicated that the target genes of DElncRNAs are enriched in ubiquitin mediated proteolysis, endocytosis and lysosome pathways. qRT-PCR validated the differential expression of several lncRNAs and miRNAs. Our results suggested that DElncRNAs participate in host response to BmNPV infection via interactions with their target genes and miRNAs. Our results will help us to improve our understanding of lncRNA-mediated regulatory roles in BmNPV infection and provide new insights into silkworm-pathogen interactions.

BMC Genomics ; 20(1): 736, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31615392


BACKGROUND: Bombyx mori nucleopolyhedrosis virus (BmNPV) is a major pathogen that threatens the sustainability of the sericultural industry. DNA methylation is a widespread gene regulation mode in epigenetics, which plays an important role in host immune response. Until now, little has been known about epigenetic regulation on virus diseases in insects. This study aims to explore the role of DNA methylation in BmNPV proliferation. RESULTS: Inhibiting DNA methyltransferase (DNMT) activity of silkworm can suppress BmNPV replication. The integrated analysis of transcriptomes and DNA methylomes in silkworm midguts infected with or without BmNPV showed that both the expression pattern of transcriptome and DNA methylation pattern are changed significantly upon BmNPV infection. A total of 241 differentially methylated regions (DMRs) were observed in BmNPV infected midguts, among which, 126 DMRs were hyper-methylated and 115 DMRs were hypo-methylated. Significant differences in both mRNA transcript level and DNA methylated levels were found in 26 genes. BS-PCR validated the hypermethylation of BGIBMGA014008, a structural maintenance of chromosomes protein gene in the BmNPV-infected midgut. In addition, DNMT inhibition reduced the expression of inhibitor of apoptosis family genes, iap1 from BmNPV, Bmiap2, BmSurvivin1 and BmSurvivin2. CONCLUSION: Our results indicate that DNA methylation plays positive roles in BmNPV proliferation and loss of DNMT activity could induce the apoptosis of infected cells to suppress BmNPV proliferation. Our results may provide a new idea and research direction for the molecular mechanism on insect-virus interaction.

Bombyx/virologia , Metilação de DNA , Inibidores Enzimáticos/farmacologia , Perfilação da Expressão Gênica/veterinária , /fisiologia , Animais , Bombyx/genética , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , DNA (Citosina-5-)-Metiltransferases/antagonistas & inibidores , Epigênese Genética , Epigenômica , Regulação da Expressão Gênica , Interações entre Hospedeiro e Microrganismos , Análise de Sequência de RNA , Replicação Viral/efeitos dos fármacos
J Proteomics ; 203: 103379, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31102755


Heat shock protein 90, an essential chaperone responsible for the correct maturation of key proteins, has been confirmed to facilitate Bombyx mori nucleopolyhedrovirus (BmNPV) proliferation but the mechanism is not clear. In this study, we use quantitative proteomics analysis to investigate the mechanism of Hsp90 in BmNPV replication. In total, 195 differentially expressed proteins (DEPs) were identified with 136 up-regulated proteins and 59 down-regulated proteins. The protein expression level of small heat shock proteins, immune-related proteins, cellular DNA repair-related proteins and zinc finger proteins is significantly enhanced while that of protein kinases is declined. KEGG pathway analysis reveals that DEPs are involved in longevity regulating pathway, mTOR signaling pathway, FoxO signaling pathway and Toll and Imd signaling pathway. Based on the DEPs results, we speculate that inhibition of Hsp90 suppresses the BmNPV infection may because it could not only stimulate the host innate immune, induce small heat shock proteins expression to maintain the cellular proteostasis but activate host transcription factors to bind to virus DNA or protein and subsequently hinder virus replication. The results will help understand the roles of Hsp90 in BmNPV infection and shed light on new clue to illustrate the molecular mechanism of silkworm-virus interaction. SIGNIFICANCE: This is the first report on Hsp90 roles in BmNPV infection based on proteomic analysis. Our findings may provide new clue and research orientation to illustrate the molecular mechanism of silkworm-virus interaction and a set of BmHsp90 candidate clients, which may involve in BmNPV infection in BmN cells.

Animals (Basel) ; 9(5)2019 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-31064075


Mulberry leaves have been used as a protein source in replacing concentrates of domestic animals, however, little is known about the relationship between supplementation level and the development of rumen epithelium. This experiment aimed to investigate the effects of different proportions of mulberry leaf powder (MLP) in dietary concentrate on rumen fermentation and rumen epithelium morphology in fattening Hu sheep. Forty three-month-old male Hu sheep with an initial body weight of 16.5 ± 0.6 kg (BW ± SD) were chosen and randomly divided into five treatments: 0% (control), 15% (T15), 30% (T30), 45% (T45) and 60% (T60) of MLP in concentrate, respectively. The results showed that the dry matter intake (DMI) and average daily gain (ADG) in treatments T15 and T30 have no significant difference with respect to the control treatment, but DMI and ADG in treatments T45 and T60 were lower than the control treatment (p < 0.05). The apparent digestibility of organic matter (OM) and neutral detergent fiber (NDF) increased linearly and quadraticly as MLP supplementation increased (p < 0.05). The concentration of ammonia (NH3-N) trended to decrease linearly with the increase of MLP supplementation (p < 0.1), whereas the microbial protein (MCP) concentration increased linearly as MLP supplementation increased (p < 0.05). In the results of rumen epithelium morphology, the width of stratum corneum was reduced, whereas the width of ruminal papillae increased (p < 0.05), and the width of stratum granulosum and stratum basale also increased as MLP increased. In summary, MLP supplementation could improve nutrient digestibility, the development of rumen papillae and stratum basale. However, high content MLP (45%-60%) supplementation decreased the growth and food intake performance of fattening Hu sheep. Therefore, 30% MLP is recommended to supplement in concentrate for fattening Hu sheep.

Mol Immunol ; 109: 134-139, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30947109


Bombyx mori nucleopolyhedrovirus (BmNPV) is a major pathogen that threatens the growth and sustainability of the sericulture industry. Accumulating studies in recent years suggest that insect viruses infection can change the host microRNAs (miRNAs) expression profile and both cellular and viral miRNAs play roles in host-pathogen interactions. Until now, the functional analysis of miRNA encoded by silkworm for host-virus interaction is limited. In this study, we validate the down-regulation of bmo-miR-2819 upon BmNPV infection by qRT-PCR and confirm the BmNPV immediately early 1 gene, ie-1 is one of the targets for bmo-miR-2819 based on the results of dual luciferase report assay. Overexpression of bmo-miR-2819 can significantly decline the abundance of IE-1 protein level in BmNPV-infected silkworm larvae. Further, the expression level of polyhedrin gene and VP39 protein of BmNPV in the infected larvae after applying bmo-miR-2819 mimics was significantly decreased comparing with that of larvae with mimic control. Our results suggest that overexpression of bmo-miR-2819 could suppress BmNPV replication by down-regulating the expression of BmNPV ie-1 gene, which demonstrate that cellular miRNAs could affect virus infection by regulating the expression of virus genes.

Bombyx/genética , Bombyx/virologia , Genes Virais , MicroRNAs/genética , /fisiologia , Replicação Viral , Animais , Sequência de Bases , Linhagem Celular , Regulação para Baixo/genética , MicroRNAs/metabolismo , Reprodutibilidade dos Testes