Quantitative detection of fungi by molecular methods: A case study on Fusarium

Abstract

The determination of fungal biomass in diverse samples plays a key role for questions in the fields of plant pathology and agriculture. Until a decade ago, morphological strain determination and quantification by agar-plating methods were the only techniques to quantify fungal infections. These methods were elaborate and time consuming and the obtained results might not always reflect the biological situation. At the end of the 1990s, numerous groups all over the world started with the molecular characterization of the genus Fusarium and defined several diagnostic sequences in the genome of the most prominent Fusarium species as suitable for the discrimination of isolates. Based on these characteristic sequences originally applied for taxonomic studies, quantitative PCR assays were developed from the turn of the millennium until now. PCR tests for certain species were also developed as well as tests for whole groups producing a particular class of toxins. Until now real-time PCR based Fusarium determinations are applied predominantly in niches in agro-biotechnology. However, to further disseminate the inexpensive and rapid quantitative PCR, the quality of analysis has to be guaranteed by defining several standards concerning the PCR procedure from DNA isolation to data analysis. Additionally, plant breeders and agronomists are familiar with toxin analysis and visual rating systems. So change in people's mind is necessary to realize the benefits of a novel technique. © 2010 Springer-Verlag Berlin Heidelberg.