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Transcriptome-wide analysis of the difference of alternative splicing in susceptible and resistant silkworm strains after BmNPV infection.

ABSTRACT: Novel alternative splicing events were identified from BmNPV-susceptible and -resistant silkworm strains after BmNPV infection using high-throughput RNA-sequencing strategy. In total, 12.82 Gb clean RNA-seq data were generated for the two midgut samples from BmNPV-susceptible and -resistant silkworm strains, and 14.78 Gb clean data for the two fat body samples. The number of alternative splicing events and isoforms in the BmNPV-susceptible silkworm strain was more than that in the BmNPV-resistant silkworm strain. Furthermore, alternative splicing genes uniquely present in BmNPV-resistant silkworm strain were involved in functions about ribosome, whereas, alternative splicing genes uniquely present in BmNPV-susceptible silkworm strain were implicated in functions like DNA helicase activity and signal transduction. Additionally, 33 expressed SR or SR-like proteins were identified, and three genes encoding SR or SR-like proteins (tetratricopeptide repeat protein 14 homolog, ubiquitin carboxyl-terminal hydrolase 32 and zinc finger CCCH domain-containing protein 18) have a higher number of different alternative splicing events between two silkworm strains. The present study suggested BmNPV treatment may have a smaller effect on the mRNA transcription in BmNPV-resistant silkworms than that in BmNPV-susceptible silkworms, and functions of alternative splicing genes are different between the two silkworm strains.

SUBMITTER: Li G

PROVIDER: S-EPMC6434002 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature

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Transcriptome-wide analysis of the difference of alternative splicing in susceptible and resistant silkworm strains after BmNPV infection.

Li Gang G Zhou Kaiyue K Zhao Guodong G Qian Heying H Xu Anying A

3 Biotech 20190325 4

Novel alternative splicing events were identified from BmNPV-susceptible and -resistant silkworm strains after BmNPV infection using high-throughput RNA-sequencing strategy. In total, 12.82 Gb clean RNA-seq data were generated for the two midgut samples from BmNPV-susceptible and -resistant silkworm strains, and 14.78 Gb clean data for the two fat body samples. The number of alternative splicing events and isoforms in the BmNPV-susceptible silkworm strain was more than that in the BmNPV-resistan ...[more]

PMID: 30944799

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Project description:Bombyx mori nucleopolyhedrovirus (BmNPV) is a major pathogen causing severe economic loss. Previous studies have revealed that some proteins in silkworm digestive juice show antiviral activity. In this study, antiviral activity examination of different resistant strains showed that the digestive juice of the resistant strain (A35) had higher inhibition to virus than the susceptible strain (P50). Subsequently, the label-free quantitative proteomics was used to study the midgut digestive juice response to BmNPV infection in P50 and A35 strains. A total of 98 proteins were identified, of which 80 were differentially expressed proteins (DEPs) with 54 enzymes and 26 nonenzymatic proteins by comparing the proteomes of infected and non-infected P50 and A35 silkworms. These DEPs are mainly involved in metabolism, proteolysis, neuroactive ligand receptor interaction, starch and sucrose metabolism and glutathione metabolism. After removing the genetic background and individual immune stress response proteins, 9 DEPs were identified potentially involved in resistance to BmNPV. Further studies showed that a serine protease, an alkaline phosphatase and serine protease inhibitor 2 isoform X1 were differentially expressed in A35 compared to P50 or post BmNPV infection. Taken together, these results provide insights into the potential mechanisms for silkworm digestive juice to provide resistance to BmNPV infection. Signifcance: Bombyx mori nucleopolyhedrovirus (BmNPV) is highly pathogenic, which has a great impact on the sericulture. BmNPV entered the midgut lumen and exposed to digestive juices after oral infection. Previous studies have revealed that some proteins in silkworm digestive juice show antiviral activity, however, current information on the digestive juice proteome of high resistant silkworm strain after BmNPV challenge compared to susceptible strain is incomprehensive. Here, we combined label-free quantification method, bioinformatics, RT-qPCR and western blot analysis and found that BmNPV infection causes some protein changes in the silkworm midgut digestive juice. The DEPs were identified in the digestive juices of different resistant strains following BmNPV infection, and screened out some proteins potentially related to resistance to BmNPV. Three important differentially expression proteins were validated by independent approaches. These findings uncover the potential role of silkworm digestive juice in providing resistance to BmNPV and supplemented the profile of the proteome of the digestive juices in B. mori.

Project description:BACKGROUND:Asian corn borer (ACB), Ostrinia furnacalis (Guenée), is the major insect pest of maize in China and countries of East and Southeast Asia, the Pacific and Australasia. ACB can develop strong resistance to the transgenic Bt maize expressing Cry1Ab, the most widely commercialized Bt maize worldwide. However, the molecular basis for the resistance mechanisms of ACB to Cry1Ab remained unclear. Two biological replicates of the transcriptome of Bt susceptible (ACB-BtS) and Cry1Ab resistant (ACB-AbR) strains of ACB were sequenced using Solexa/Illumina RNA-Seq technology to identify Cry1Ab resistance-relevant genes. RESULTS:The numbers of unigenes for two biological replications were 63,032 and 53,710 for ACB-BtS and 57,770 and 54,468 for ACB-AbR. There were 35,723 annotated unigenes from ACB reads found by BLAST searching NCBI non-redundant, NCBI non-redundant nucleotide, Swiss-prot protein, Kyoto Encyclopedia of Genes and Genomes, Cluster of Orthologous Groups of proteins, and Gene Ontology databases. Based on the NOISeq method, 3,793 unigenes were judged to be differentially expressed between ACB-BtS and ACB-AbR. Cry1Ab resistance appeared to be associated with change in the transcription level of enzymes involved in growth regulation, detoxification and metabolic/catabolic process. Among previously described Bt toxin receptors, the differentially expressed unigenes associated with aminopeptidase N and chymotrypsin/trypsin were up-regulated in ACB-AbR. Whereas, other putative Cry receptors, cadherin-like protein, alkaline phosphatase, glycolipid, actin, V-type proton ATPase vatalytic, heat shock protein, were under-transcripted. Finally, GPI-anchor biosynthesis was found to be involved in the significantly enriched pathway, and all genes mapped to the pathway were substantially down-regulated in ACB-AbR. CONCLUSION:To our knowledge, this is the first comparative transcriptome study to discover candidate genes involved in ACB Bt resistance. This study identified differentially expressed unigenes related to general Bt resistance in ACB. The assembled, annotated transcriptomes provides a valuable genomic resource for further understanding of the molecular basis of ACB Bt resistance mechanisms.

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Dataset Information

Transcriptome-wide analysis of the difference of alternative splicing in susceptible and resistant silkworm strains after BmNPV infection.

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Transcriptome-wide analysis of the difference of alternative splicing in susceptible and resistant silkworm strains after BmNPV infection.

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