Identification of stage-specific markers in Trichoderma atrovide and their use in ecological studies

Several species of Trichoderma are fungal biocontrol agents that are active against a wide range of soil-borne, plant-pathogenic fungi in the glasshouse and field. However, the ecological fitness and hence efficacy of Trichoderma spp. as biocontrol agents in natural soils can be limited by the variable influence of biotic and abiotic factors. In the soil, fungi are present as both actively growing organisms and as dormant propagules. Thus, the use of stage-specific molecular markers to determine development and differentiation in Trichoderma provides a particularly useful tool for studying the influence of ecological factors on the life cycle of the fungus. Here we present a subtractive/suppressive hybridization analysis of genes regulated during critical morphological stages in T. atroviride LU132. The validation of the cDNA libraries was performed by Northern blot and by quantitative real-time reverse transcription-PCR (qRT-PCR). In addition, conidia of Trichoderma LU132 were inoculated in sterile soil, maintained under glasshouse conditions and collected at different time points. The influence of Trichoderma on maize plant development will be discussed. We consider that the use of stage specific markers will significantly increase our understanding of the ecological niche of Trichoderm