“…Wang et al (2008) reported that TIF-4A is highly stable in silkworms; therefore, we did not perform stability analysis in the present study. Additionally, while A3 is a commonly used RG in silkworm studies (Bao et al 2008;Gao et al 2014a, b;Wang et al 2014Wang et al , 2015Kolliopoulou et al 2015), our study showed that A3 was affected by viral infection and HT stress. Consistent with this, Bao et al (2009) reported that A3 levels are altered after BmNPV infection in silkworms.…”
Section: Discussionmentioning
confidence: 48%
“…Viruses and HTs are serious threats to sericulture (Jiang and Xia 2014;Wang et al 2014); therefore, exploring the responses of silkworms to these stresses is critical (Bao et al 2008;Xue et al 2012;Gao et al 2014a;Li et al 2014;Wang et al 2014;Jiang et al, unpublished data). qPCR is a common tool for analysis of the expression levels of stressresponsive genes, and identification of appropriate RGs is critical for data normalization in qPCR (Huggett et al 2005;Wang et al 2008;Velada et al 2014).…”
Section: Discussionmentioning
confidence: 99%
“…Studies of the response of silkworms to viruses and HTs are the foundation for the cultivation of resistant varieties. Microarrays (Wu et al 2011;Zhou et al 2013), suppression subtractive hybridization (Bao et al 2008(Bao et al , 2009(Bao et al , 2010, and RNA-seq (Xue et al 2012;Gao et al 2014b;Guo et al 2015;Kolliopoulou et al 2015;Wang et al 2015) have been used for the analysis of stress-responsive genes in silkworms after viral challenge. RNA-seq has also been used for gene expression analysis in silkworms after HT stress (Li et al 2014;Wang et al 2014).…”
Section: Introductionmentioning
confidence: 99%
“…BmNPVGAPDH(Bao et al 2009(Bao et al , 2010, 18S rRNA(Xue et al 2012), TIF-3 (Zhou et al 2013), A3 (Wang et al 2015), TIF-4A (Jiang et al 2012a, b, 2013b) BmCPV A3 (Gao et al 2014a, b; Kolliopoulou et al 2015)BmDNV-Z A3(Bao et al 2008), GAPDH(Bao et al 2013) HT A1(Li et al 2014), A3(Wang et al 2014) …”
Viruses and high temperature (HT) are the primary threats to silkworms. Changes in the expression of stress-response genes can be measured using quantitative polymerase chain reaction (qPCR) after exposure to viruses or HT. However, appropriate reference genes (RGs) for qPCR data normalization have not been established in this organism. In this study, we summarized the RGs used in the previous silkworm studies after infection with Bombyx mori nucleopolyhedrovirus (BmNPV), B. mori cytoplasmic polyhedrosis virus (BmCPV), or B. mori densovirus (BmDNV) or after HT treatment. The expression levels of these RGs were extracted from silkworm transcriptome data to screen for candidate RGs that were unaffected by the experimental conditions. Actin-1 (A1), actin-3 (A3), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation initiation factor 4a (TIF-4A) were selected for further qPCR verification. The results of RNA-seq and qPCR showed that GAPDH and TIF-4A were suitable RGs after BmNPV challenge or HT stress, whereas TIF-4A was an appropriate RG for BmCPV or BmDNV-Z challenge in silkworms. These results suggested that TIF-4A may be the most appropriate RG for gene expression analysis after challenge with viruses or HT in silkworms.
“…Wang et al (2008) reported that TIF-4A is highly stable in silkworms; therefore, we did not perform stability analysis in the present study. Additionally, while A3 is a commonly used RG in silkworm studies (Bao et al 2008;Gao et al 2014a, b;Wang et al 2014Wang et al , 2015Kolliopoulou et al 2015), our study showed that A3 was affected by viral infection and HT stress. Consistent with this, Bao et al (2009) reported that A3 levels are altered after BmNPV infection in silkworms.…”
Section: Discussionmentioning
confidence: 48%
“…Viruses and HTs are serious threats to sericulture (Jiang and Xia 2014;Wang et al 2014); therefore, exploring the responses of silkworms to these stresses is critical (Bao et al 2008;Xue et al 2012;Gao et al 2014a;Li et al 2014;Wang et al 2014;Jiang et al, unpublished data). qPCR is a common tool for analysis of the expression levels of stressresponsive genes, and identification of appropriate RGs is critical for data normalization in qPCR (Huggett et al 2005;Wang et al 2008;Velada et al 2014).…”
Section: Discussionmentioning
confidence: 99%
“…Studies of the response of silkworms to viruses and HTs are the foundation for the cultivation of resistant varieties. Microarrays (Wu et al 2011;Zhou et al 2013), suppression subtractive hybridization (Bao et al 2008(Bao et al , 2009(Bao et al , 2010, and RNA-seq (Xue et al 2012;Gao et al 2014b;Guo et al 2015;Kolliopoulou et al 2015;Wang et al 2015) have been used for the analysis of stress-responsive genes in silkworms after viral challenge. RNA-seq has also been used for gene expression analysis in silkworms after HT stress (Li et al 2014;Wang et al 2014).…”
Section: Introductionmentioning
confidence: 99%
“…BmNPVGAPDH(Bao et al 2009(Bao et al , 2010, 18S rRNA(Xue et al 2012), TIF-3 (Zhou et al 2013), A3 (Wang et al 2015), TIF-4A (Jiang et al 2012a, b, 2013b) BmCPV A3 (Gao et al 2014a, b; Kolliopoulou et al 2015)BmDNV-Z A3(Bao et al 2008), GAPDH(Bao et al 2013) HT A1(Li et al 2014), A3(Wang et al 2014) …”
Viruses and high temperature (HT) are the primary threats to silkworms. Changes in the expression of stress-response genes can be measured using quantitative polymerase chain reaction (qPCR) after exposure to viruses or HT. However, appropriate reference genes (RGs) for qPCR data normalization have not been established in this organism. In this study, we summarized the RGs used in the previous silkworm studies after infection with Bombyx mori nucleopolyhedrovirus (BmNPV), B. mori cytoplasmic polyhedrosis virus (BmCPV), or B. mori densovirus (BmDNV) or after HT treatment. The expression levels of these RGs were extracted from silkworm transcriptome data to screen for candidate RGs that were unaffected by the experimental conditions. Actin-1 (A1), actin-3 (A3), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation initiation factor 4a (TIF-4A) were selected for further qPCR verification. The results of RNA-seq and qPCR showed that GAPDH and TIF-4A were suitable RGs after BmNPV challenge or HT stress, whereas TIF-4A was an appropriate RG for BmCPV or BmDNV-Z challenge in silkworms. These results suggested that TIF-4A may be the most appropriate RG for gene expression analysis after challenge with viruses or HT in silkworms.
“…Thus the BC 6 strain also carries the recessive gene (nsd-Z/nsd-Z) . In theory, the inherited background of BC 6 has 99.9% identity with Huaba35, the susceptible strain [10] .…”
Bombyx mori densonucleosis virus (BmDNV) is one of the most disastrous viruses in cocoon production. Silkworm resistance to BmDNV has been examined previously using a number of traditional biochemical and molecular techniques. In this study, a near isogenic line, BC6, was constructed to eliminate the difference in inherited background, which has 99.9% identity with the susceptible strain but carries a resistant gene. We utilized a proteomic approach involving two-dimensional differential gel electrophoresis and mass spectrometry to examine changes in the midgut proteins from the susceptible and resistant silkworm larvae infected with BmDNV. The protein profiles were compared and 9 differentially expressed proteins were identified by mass spectrometry. In the resistant strains, the heat-shock 70-kDa protein cognate, cytochrome P450, vacuolar ATP synthase subunit B, arginine kinase, vacuolar ATP synthase subunit D and glutathione S-transferase sigma were strongly upregulated and α-tubulin was downregulated. Our results imply that these upregulated genes and the downregulated genes might be involved in B. mori immune responses against BmDNV-Z infection.
Alkaline trypsin protein of molecular mass 25,436 Da purified from the digestive juice of Bombyx mori larvae indicated strong antiviral activity against Bombyx mori nucleopolyhedrovirus (BmNPV) under in vitro conditions. Partial N-terminal amino acid sequence of the protein was determined and the cDNA was cloned based on the amino acid sequence. A homology search of the deduced amino acid sequence of the cDNA showed 55% identity with Helicoverpa armigera trypsin and the active site of this protein was completely conserved. Hence, the protein was designated B. mori trypsin (Bmtryp). The results suggest that Bmtryp, an insect digestive enzyme, can be a potential antiviral factor against BmNPV at the initial site of viral infection.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.