Identification of differentially expressed genes from Trichoderma harzianum during growth on cell wall of Fusarium solanias a tool for biotechnological application

Abstract

BackgroundThe species of T. harzianum are well known for their biocontrol activity against many plant pathogens. However, there is a lack of studies concerning its use as a biological control agent against F. solani, a pathogen involved in several crop diseases. In this study, we have used subtractive library hybridization (SSH) and quantitative real-time PCR (RT-qPCR) techniques in order to explore changes in T. harzianum genes expression during growth on cell wall of F. solani (FSCW) or glucose. RT-qPCR was also used to examine the regulation of 18 genes, potentially involved in biocontrol, during confrontation between T. harzianum and F. solani.ResultsData obtained from two subtractive libraries were compared after annotation using the Blast2GO suite. A total of 417 and 78 readable EST sequence were annotated in the FSCW and glucose libraries, respectively. Functional annotation of these genes identified diverse biological processes and molecular functions required during T. harzianum growth on FSCW or glucose. We identified various genes of biotechnological value encoding to proteins which function such as transporters, hydrolytic activity, adherence, appressorium development and pathogenesis. Fifteen genes were up-regulated and sixteen were down-regulated at least at one-time point during growth of T. harzianum in FSCW. During the confrontation assay most of the genes were up-regulated, mainly after contact, when the interaction has been established.ConclusionsThis study demonstrates that T. harzianum expressed different genes when grown on FSCW compared to glucose. It provides insights into the mechanisms of gene expression involved in mycoparasitism of T. harzianum against F. solani. The identification and evaluation of these genes may contribute to the development of an efficient biological control agent.

Identification of mycoparasitism-related genes against the phytopathogen Sclerotinia sclerotiorum through transcriptome and expression profile analysis in Trichoderma harzianum

“…This fact is consistent with the extensive metabolic activity expected for a filamentous fungus growing on a rich medium (glucose 2% medium) with an easily assimilable substrate [17]. Under this culture condition up-regulation of a specific subset of oxidoreductases and nucleotide binding proteins endoding genes related to primary metabolism is commonly observed for Trichoderma species, which are down-regulated in the presence of complex substrates [13]. Vieira et al in 2013 showed the same pattern of repressed categories when T. harzianum was grown on Fusarium solani cell wall, suggesting that this result is not pathogen-dependent.…”

Evaluating the mycostimulation potential of select carbon amendments for the degradation of a model PAH by an ascomycete strain enriched from a superfund site

“…Primers were designed with redundancy to complement the identified conserved sequence homology that spanned between 100 and 150 bp in order to account for strain-strain differences. Primer sets for chitinase 33 , were chosen because it is a chitinase that lacks a substrate-binding domain and has been shown to be involved in mycoparasitism [30, 31]. The primer set for qid74 was selected because it was shown to mediate hydrophobic surface attachment during mycoparasitism [32].…”

“…Primer sets for chitinase 33 , were chosen because it is a chitinase that lacks a substrate-binding domain and has been shown to be involved in mycoparasitism [30, 31]. The primer set for qid74 was selected because it was shown to mediate hydrophobic surface attachment during mycoparasitism [32]. For quantification purposes, each target was normalized to the 18S gene using previously published methods [33].…”

Subtilisin-like serine protease gene <i>TghSS42</i> from <i>Trichoderma ghanense</i> ACCC 30153 was successfully expressed in <i>Escherichia coli</i> and recombinant protease rTghSS42 exhibited antifungal ability to five phytopathogens

“…The subtilisin serine protease gene Spm 1 from T. asperellum T4 was all up-regulated in response to different fungal phytopathogen induction Liu et al, 2010 . The transcription of serine protease gene in T. harzianum was up-regulated under Fusariura solani cell walls treatment Vieira et al, 2013 . Northern blot analysis indicated that the serine protease gene SL41 from T. harzianum T88 could be induced by phytopathogenic fungi cell walls and the protease SL41 from T. harzianum T88 exerted broad-spectrum antifungal activity against phytopathogenic fungi Liu and Yang, 2013 .…”