Identification of mycoparasitism-related genes against the phytopathogen Sclerotinia sclerotiorum through transcriptome and expression profile analysis in Trichoderma harzianum
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2014
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Background: The species of T. harzianum are well known for their biocontrol activity against plant pathogens.
However, few studies have been conducted to further our understanding of its role as a biological control agent
against S. sclerotiorum, a pathogen involved in several crop diseases around the world. In this study, we have used
RNA-seq and quantitative real-time PCR (RT-qPCR) techniques in order to explore changes in T. harzianum gene
expression during growth on cell wall of S. sclerotiorum (SSCW) or glucose. RT-qPCR was also used to examine
genes potentially involved in biocontrol, during confrontation between T. harzianum and S. sclerotiorum.
Results: Data obtained from six RNA-seq libraries were aligned onto the T. harzianum CBS 226.95 reference genome
and compared after annotation using the Blast2GO suite. A total of 297 differentially expressed genes were found
in mycelia grown for 12, 24 and 36 h under the two different conditions: supplemented with glucose or SSCW.
Functional annotation of these genes identified diverse biological processes and molecular functions required
during T. harzianum growth on SSCW or glucose. We identified various genes of biotechnological value encoding
proteins with functions such as transporters, hydrolytic activity, adherence, appressorium development and pathogenesis.
To validate the expression profile, RT-qPCR was performed using 20 randomly chosen genes. RT-qPCR expression profiles
were in complete agreement with the RNA-Seq data for 17 of the genes evaluated. The other three showed differences
at one or two growth times. During the confrontation assay, some genes were up-regulated during and after contact,
as shown in the presence of SSCW which is commonly used as a model to mimic this interaction.
Conclusions: The present study is the first initiative to use RNA-seq for identification of differentially expressed genes
in T. harzianum strain TR274, in response to the phytopathogenic fungus S. sclerotiorum. It provides insights into the
mechanisms of gene expression involved in mycoparasitism of T. harzianum against S.sclerotiorum. The RNA-seq data
presented will facilitate improvement of the annotation of gene models in the draft T. harzianum genome and provide
important information regarding the transcriptome during this interaction.
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T. harzianum, Mycoparasitism, S. sclerotiorum, RNA-seq, Gene expression
Citação
STEINDORFF, Andrei Stecca; RAMADA, Marcelo Henrique Soller; COELHO, Alexandre Siqueira Guedes; MILLER, Robert Neil Gerard; PAPPAS JUNIOR, Georgios Joannis; ULHOA, Cirano José; NORONHA, Eliane Ferreira. Identification of mycoparasitism-related genes against the phytopathogen Sclerotinia sclerotiorum through transcriptome and expression profile analysis in Trichoderma harzianum. BMC Genomics, London, v. 15, n. 204, p. 1-14, 2014.