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.

Highlights

  • The species of T. harzianum are well known for their biocontrol activity against many plant pathogens

  • Identification of differentially expressed genes during growth of T. harzianum in FSCW or glucose In this study, a suppression subtractive hybridization (SSH) approach, which is an efficient method for the isolation of differentially expressed genes, was used to isolate and identify genes that are differentially expressed during T. harzianum growth on FSCW or glucose

  • In order to obtain a cDNA library enriched for sequences representative of those genes up-regulated in the presence of FSCW-library, cDNA from T. harzianum grown in FSCW was used as the tester and cDNA from T. harzianum grown in glucose medium as the driver (Glc-library)

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Summary

Introduction

The species of T. harzianum are well known for their biocontrol activity against many plant pathogens. There is a lack of studies concerning its use as a biological control agent against F. solani, a pathogen involved in several crop diseases. Species of the genus Trichoderma are widely known for their biotechnological interest, their use as biocontrol agents requires a comprehensive analysis of the biological principles of their action. Comprehensive analysis of the molecular mechanisms used by T. harzianum during interaction with F. solani is required in order to identify the molecular determinants of its role as a biological control agent [10,11]. The identification of the Trichoderma genes involved in these mechanisms and analysis of their expression profiles can provide researchers with biotechnological tools that exhibit anti-fungal activity and that could potentially be used as transgenes capable of inducing resistance to pathogens in economically valuable plants

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