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. 2016 Nov 7;11(11):e0165959.
doi: 10.1371/journal.pone.0165959. eCollection 2016.

Transcriptome Analysis of the Midgut of the Chinese Oak Silkworm Antheraea pernyi Infected with Antheraea pernyi Nucleopolyhedrovirus

Xi-Sheng Li  1   2   3 , Guo-Bao Wang  2 , Ying Sun  1   2 , Wei Liu  2 , Ying-Zi He  3 , Feng-Cheng Wang  3 , Yi-Ren Jiang  2 , Li Qin  1   2
Affiliations

Transcriptome Analysis of the Midgut of the Chinese Oak Silkworm Antheraea pernyi Infected with Antheraea pernyi Nucleopolyhedrovirus

Xi-Sheng Li et al. PLoS One. .

Abstract

The Antheraea pernyi nucleopolyhedrovirus (ApNPV) is an exclusive pathogen of A. pernyi. The intense interactions between ApNPV and A. pernyi cause a series of physiological and pathological changes to A. pernyi. However, no detailed report exists regarding the molecular mechanisms underlying the interactions between ApNPV and A. pernyi. In this study, four cDNA libraries of the A. pernyi midgut, including two ApNPV-infected groups and two control groups, were constructed for transcriptomic analysis to provide new clues regarding the molecular mechanisms that underlie these interactions. The transcriptome of the A. pernyi midgut was de novo assembled using the Trinity platform because of the lack of a genome resource for A. pernyi. Compared with the controls, a total of 5,172 differentially expressed genes (DEGs) were identified, including 2,183 up-regulated and 2,989 down-regulated candidates, of which 2,965 and 911 DEGs were classified into different GO categories and KEGG pathways, respectively. The DEGs involved in A. pernyi innate immunity were classified into several categories, including heat-shock proteins, apoptosis-related proteins, serpins, serine proteases and cytochrome P450s. Our results suggested that these genes were related to the immune response of the A. pernyi midgut to ApNPV infection via their essential roles in regulating a variety of physiological processes. Our results may serve as a basis for future research not only on the molecular mechanisms of ApNPV invasion but also on the anti-ApNPV mechanism of A. pernyi.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Volcano plot of the DEGs.
The horizontal ordinate represents the fold change of gene expression in the different experimental groups, and the vertical ordinate represents the statistical significance of the change of gene expression. Each point in the plot represents each gene, and the red and green points represent the significant up- and down-regulated genes.
Fig 2
Fig 2. Pearson correlation between the samples in the Ap_CK and Ap_NPV groups.
Fig 3
Fig 3. GO analysis of the DEGs.
Genes with differential expression levels were classified as biological process, cellular component, and molecular function by WEGO according to the GO terms. The numbers of genes mapped to the GO terms are provided in the left panel.
Fig 4
Fig 4. KEGG pathway analysis of the DEGs.
The pathways were clustered into metabolism, genetic information processing, environmental information processing and cellular processes. The left panel lists the numbers of genes that mapped to each of the pathways.
Fig 5
Fig 5. The top 20 pathways of the DEGs clustered by KEGG analysis.
(A) The top 20 pathways for the down-regulated genes. (B) The top 20 pathways for the up-regulated genes.
Fig 6
Fig 6. Quantitative RT-PCR analysis was used to validate the differentially expressed genes according to RNA-seq.
X-axis represents the eight genes selected for qRT-PCR validation. comp45066_c0, heat shock protein 60 (Chilo suppressalis); comp42863_c0, heat shock protein 19.9 (Bombyx mori); comp63819_c0, apoptosis-inducing factor 3 (Homo sapiens); comp18400_c0, serine protease 3 (Lonomia obliqua); comp44655_c0, serine protease 5 (Mamestra configurata); comp32864_c0, serine protease inhibitor 12 (Bombyx mori); comp39389_c0, serine protease inhibitor 5 (Bombyx mori); comp37472_c0, cytochrome CYP324A1 (Spodoptera littoralis). Y-axis represents the relative expression level. The Ct values of each reaction were normalized to the endogenous control actin3.
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Grants and funding

This work was supported by the National Modern Agriculture Industry Technology System Construction Project (Silkworm and Mulberry) and the Magnitude Science and Technology Projects of Liaoning Province to LQ; the Cultivation Plan for Youth Agricultural Science and Technology Innovative Talents of Liaoning Province (2014040) and the Scientific Research Project for the Education Department of Liaoning Province (L2014255) to YRJ.

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