Abstract

BackgroundCombating the action of plant pathogenic microorganisms by mycoparasitic fungi has been announced as an attractive biological alternative to the use of chemical fungicides since two decades. The fungal genus Trichoderma includes a high number of taxa which are able to recognize, combat and finally besiege and kill their prey. Only fragments of the biochemical processes related to this ability have been uncovered so far, however.ResultsWe analyzed genome-wide gene expression changes during the begin of physical contact between Trichoderma atroviride and two plant pathogens Botrytis cinerea and Rhizoctonia solani, and compared with gene expression patterns of mycelial and conidiating cultures, respectively. About 3000 ESTs, representing about 900 genes, were obtained from each of these three growth conditions. 66 genes, represented by 442 ESTs, were specifically and significantly overexpressed during onset of mycoparasitism, and the expression of a subset thereof was verified by expression analysis. The upregulated genes comprised 18 KOG groups, but were most abundant from the groups representing posttranslational processing, and amino acid metabolism, and included components of the stress response, reaction to nitrogen shortage, signal transduction and lipid catabolism. Metabolic network analysis confirmed the upregulation of the genes for amino acid biosynthesis and of those involved in the catabolism of lipids and aminosugars.ConclusionThe analysis of the genes overexpressed during the onset of mycoparasitism in T. atroviride has revealed that the fungus reacts to this condition with several previously undetected physiological reactions. These data enable a new and more comprehensive interpretation of the physiology of mycoparasitism, and will aid in the selection of traits for improvement of biocontrol strains by recombinant techniques.

Highlights

  • Combating the action of plant pathogenic microorganisms by mycoparasitic fungi has been announced as an attractive biological alternative to the use of chemical fungicides since two decades

  • Characterization of the EST database We constructed EST libraries of T. atroviride from four different cultivation conditions: (a) confrontation with two plant pathogenic hosts (Botrytis cinerea, Rhizoctonia solani) on agar plates in the dark, using an mRNA extraction time point when their hyphae were only 1-2 mm apart; (b) a respective control consisting of only T. atroviride growing in the dark and not sporulating; (c) cultures illuminated by blue light to induce conidiation [13]; and (d) mycelia growing on plates in the dark and subjected to stress provoked by mechanical injury, a method known to lead to sporulation independent of light [13]

  • The rationale of this approach was that genes which would be uniquely expressed under (a) could so be distinguished from those required for filamentous growth and sporulation by T. atroviride

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Summary

Introduction

Combating the action of plant pathogenic microorganisms by mycoparasitic fungi has been announced as an attractive biological alternative to the use of chemical fungicides since two decades. The ability to utilize other fungi as a source of nutrients and e.g. combat the action of plant pathogenic fungi by antagonistic or mycoparasitic fungi is an attractive biological alternative to the use of chemical fungicides [3]. More global analyses (e.g. by the use of substractive hybridiation techniques, proteomics or expressed sequence tag (EST) approaches) have been performed with different Trichoderma species, but have suffered from the fact that no full genome sequences of the used species were available for an in depth interpretation of the results These studies mostly used a mix of different growth conditions to generate snapshots of the genetic arsenal of these fungi, rather than to study individual events during the mycoparasitic attack [7,8,9,10,11]

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