Characterization of natural antisense transcript, sclerotia development and secondary metabolism by strand-specific RNA sequencing of Aspergillus flavus

19Citations
Citations of this article
32Readers
Mendeley users who have this article in their library.

Abstract

Aspergillus flavus has received much attention owing to its severe impact on agriculture and fermented products induced by aflatoxin. Sclerotia morphogenesis is an important process related to A. flavus reproduction and aflatoxin biosynthesis. In order to obtain an extensive transcriptome profile of A. flavus and provide a comprehensive understanding of these physiological processes, the isolated mRNA of A. flavus CA43 cultures was subjected to high-throughput strand-specific RNA sequencing (ssRNA-seq). Our ssRNA-seq data profiled widespread transcription across the A. flavus genome, quantified vast transcripts (73% of total genes) and annotated precise transcript structures, including untranslated regions, upstream open reading frames (ORFs), alternative splicing variants and novel transcripts. We propose natural antisense transcripts in A. flavus might regulate gene expression mainly on the post-transcriptional level. This regulation might be relevant to tune biological processes such as aflatoxin biosynthesis and sclerotia development. Gene Ontology annotation of differentially expressed genes between the mycelia and sclerotia cultures indicated sclerotia development was related closely to A. flavus reproduction. Additionally, we have established the transcriptional profile of aflatoxin biosynthesis and its regulation model. We identified potential genes linking sclerotia development and aflatoxin biosynthesis. These genes could be used as targets for controlled regulation of aflatoxigenic strains of A. flavus. © 2014 Wu et al.

Figures

  • Figure 1. Summary of RNA sequencing of A. flavus CA43. (A) Matching summary of ssRNA-seq reads to the A. flavus genome. (B) Box and whisker plots of log2-transformed RPKM for exons, novel transcripts, introns and intergenic regions. Horizontal lines in boxes, the first, median and third quartile. Other horizontal lines, the inner boundaries. Diamonds, data outside the inner boundaries. (C) Mapping coverage of the transcribed A. flavus genes. doi:10.1371/journal.pone.0097814.g001
  • Figure 2. WEGO comparison of A. flavus and A. oryzae transcribed transcriptional factors. In total, 525 of the 617 A. flavus transcribed TFs and 461 of the 571 A. oryzae transcribed TFs were assigned to GO categories. doi:10.1371/journal.pone.0097814.g002
  • Figure 3. Detailed annotation of the A. flavus gene model. (A) A novel transcript (TU134) identified in contig gi|156128545|gb|AAIH02000077.1| in the forward orientation. Red bar, the novel transcript TU134 identified in the mycelia sample. Red curve, expression level (log2transformed reads count) of the transcripts located in the forward strand. Green curve, the expression level of the transcripts located in the reverse strand. (B) Scatterplot and histograms showing the length distribution of the 59- and 39-UTRs of A. flavus CA43 genes. (C) UTR and uORF illustration for an annotated A. flavus gene (AFLG2_08360). doi:10.1371/journal.pone.0097814.g003
  • Figure 4. Illustration of AS events (RI, SE, A5SS and A3SS) identified in the A. flavus transcriptome. (A) Green bar, the transcriptional active region (TAR). Orange bar, A. flavus annotated genes. Knuckle lines (black or red), the relation with TARs or exons. The types and amounts of AS events are shown. (B) Amino acid alignment and homologous 3D modeling of AFL2G_07666 (SphK) and its AS variant (SphK _variant). Broken line, the skipped exon of SphK. doi:10.1371/journal.pone.0097814.g004
  • Figure 5. NAT analysis in A. flavus CA43. (A) The relationship between the location of NAT and its corresponding genes. Downstream, Upstream and Inside, NATs located in the downstream, upstream or inside of their corresponding genes, respectively. Intersected, NAT located across one gene and its 59- or 39- flanking region. (B) Box and whisker plots of log2-transformed RPKM for A. flavus annotated genes with or without NATs. (C) Occurrence of NAT in the SdeA gene (AFL2G_00446). Orange curves, the NAT in the SdeA gene. doi:10.1371/journal.pone.0097814.g005
  • Figure 7. Differential transcription of A. flavus aflatoxin biosynthetic pathway. (A) The expression status of AF biosynthesis in the mycelia and sclerotia states. (B) The regulation model of AF biosynthesis-related genes in the mycelia and sclerotia states. The orthologs of the AF biosynthetic pathway genes are in the same order as in (A). Boxes, DEGs (p,0.001 and change .2-fold) between mycelia and sclerotia states, denoted by the names of their homologs in A. nidulans. Red and green boxes, up-regulated and down-regulated genes in the sclerotia state, respectively. White boxes, genes with no significant change between the mycelia and sclerotia states. doi:10.1371/journal.pone.0097814.g007

References Powered by Scopus

Mapping and quantifying mammalian transcriptomes by RNA-Seq

10998Citations
N/AReaders
Get full text

The functions of animal microRNAs

9742Citations
N/AReaders
Get full text

TopHat: Discovering splice junctions with RNA-Seq

9634Citations
N/AReaders
Get full text

Cited by Powered by Scopus

Association of fungal secondary metabolism and sclerotial biology

117Citations
N/AReaders
Get full text

Integrative analyses reveal transcriptome-proteome correlation in biological pathways and secondary metabolism clusters in A. flavus in response to temperature

82Citations
N/AReaders
Get full text

Global survey of canonical aspergillus flavus G Protein-Coupled receptors

63Citations
N/AReaders
Get full text

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Cite

CITATION STYLE

APA

Wu, X., Zhou, B., Yin, C., Guo, Y., Lin, Y., Pan, L., & Wang, B. (2014). Characterization of natural antisense transcript, sclerotia development and secondary metabolism by strand-specific RNA sequencing of Aspergillus flavus. PLoS ONE, 9(5). https://doi.org/10.1371/journal.pone.0097814

Readers over time

‘14‘15‘16‘17‘18‘19‘20‘21‘23‘2402468

Readers' Seniority

Tooltip

PhD / Post grad / Masters / Doc 16

70%

Researcher 5

22%

Professor / Associate Prof. 2

9%

Readers' Discipline

Tooltip

Agricultural and Biological Sciences 12

52%

Biochemistry, Genetics and Molecular Bi... 8

35%

Environmental Science 2

9%

Computer Science 1

4%

Save time finding and organizing research with Mendeley

Sign up for free
0