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, 2019, 1864168
eCollection

Transcriptome Analysis Reveals the Molecular Mechanisms Underlying Adenosine Biosynthesis in Anamorph Strain of Caterpillar Fungus

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Transcriptome Analysis Reveals the Molecular Mechanisms Underlying Adenosine Biosynthesis in Anamorph Strain of Caterpillar Fungus

Shan Lin et al. Biomed Res Int.

Abstract

Caterpillar fungus is a well-known fungal Chinese medicine. To reveal molecular changes during early and late stages of adenosine biosynthesis, transcriptome analysis was performed with the anamorph strain of caterpillar fungus. A total of 2,764 differentially expressed genes (DEGs) were identified (p ≤ 0.05, |log2 Ratio| ≥ 1), of which 1,737 were up-regulated and 1,027 were down-regulated. Gene expression profiling on 4-10 d revealed a distinct shift in expression of the purine metabolism pathway. Differential expression of 17 selected DEGs which involved in purine metabolism (map00230) were validated by qPCR, and the expression trends were consistent with the RNA-Seq results. Subsequently, the predicted adenosine biosynthesis pathway combined with qPCR and gene expression data of RNA-Seq indicated that the increased adenosine accumulation is a result of down-regulation of ndk, ADK, and APRT genes combined with up-regulation of AK gene. This study will be valuable for understanding the molecular mechanisms of the adenosine biosynthesis in caterpillar fungus.

Conflict of interest statement

The authors declare that they have no conflicts of interest.

Figures

Figure 1
Figure 1
Time course of purine accumulation. Mycelia biomass and purine contents both sharply increased from the early stage (4 d) of secondary metabolite accumulation to and late stage (10 d).
Figure 2
Figure 2
Expression analysis between early and late samples based on the expression matrix. (a) Venn analysis between early and late samples, the intersecting areas of the circles represent the number of unigene/transcript common to each group, inter-sample Venn analysis can obtain co-expression and specific expression genes/transcripts between samples and groups. (b) Inter-sample correlation analysis, different colors represent the size of the correlation coefficient between samples. Correlation analysis helps to understand the correlation between samples, especially among biological duplicates.
Figure 3
Figure 3
Volcano plot of differentially expressed genes. The abscissa is the multiple change value of gene expression difference between two samples. The ordinate is the statistical test value of the variation of gene amount. The red dots indicate significantly upregulated unigene/transcript, the green dots indicate significantly downregulated unigene/transcript, the black dots indicate no significantly expressed unigene/transcript.
Figure 4
Figure 4
GO classification statistics of DEGs. The abscissa in the figure represents the secondary classification of GO. The vertical axis represents the percentage of the total unigene/transcript contained in this secondary classification.
Figure 5
Figure 5
Histogram of KEGG pathway classification statistics. The ordinate is the name of KEGG metabolic pathway, the abscissa is the number of transcript/unigene annotated into the pathway.
Figure 6
Figure 6
Validation of DEGs which involved in purine metabolism by qPCR. Light gray columns represent the expression level of the DEGs obtained by RNA-Seq, and the dark gray columns represent the qPCR results.
Figure 7
Figure 7
The predicted adenosine biosynthesis pathway and gene expression profiles in caterpillar fungus. Graphs show RNA-Seq and qPCR expression analysis of DEGs at late stage compared with early stage of secondary metabolites biosynthesis. Genes in red font were down-regulated and in green font were up-regulated, with asterisks (∗) indicating significant differences (p < 0.05).

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