Live-cell imaging in Trichoderma

Abstract Twenty year ago, Trichoderma was among the first filamentous fungi that successfully utilized the green fluorescent protein, sGFP, for cell biological research. Since then, several fluorescent proteins (FPs) have been applied as visual reporters of fungal biomass, gene expression, and subcellular protein localization. This chapter reviews the live-cell imaging applications of FPs established in various Trichoderma species so far and highlights key advances achieved with this technology. Furthermore, the practical aspects regarding the application of fluorescent dyes, the choice of microscopy technology, and the optimization of cultivation and sample preparation methods for live-cell imaging are discussed. The complexity and quality of live-cell imaging analyses conducted with Trichoderma is at state-of-the-art level. The impact of previous and recent hallmark studies is an encouragement to further advance live-cell imaging analysis in this exciting and diverse fungal genus operating in biotechnologically and agriculturally highly relevant areas.