• Fusarium mycotoxins.
• Zearalenone
• Fungicides
• Reference genes
• Absolute quantification
• Relative quantification
• RT-qPCR
• NormFinder
• BestKeeper
• MFS transporters
• ABC transporters
• Clonostachys rosea

The aim of this work was to provide a validation of 6 putative reference genes that could be included in a gene expression study related to the beneficial isolate Clonostachys rosea IK726. No data are available about validation of reference genes in C. rosea. In the present work the selection of these genes was based both on published data referred to other biological systems and on the available Expressed Sequence Tags (ESTs) from C. rosea IK726. Several growth conditions were taken into account for C. rosea IK726, whose mRNA was extracted and retro-transcribed into cDNA and then used as template for Real Time quantitative PCR (RT-qPCR). Two different excel based algorithms – ‘NormFinder’ and ‘BestKeeper’ – were chosen in order to analyze expression data and find the ‘best’ reference gene among chosen ones. From statistical analysis, actin was chosen and used as reference gene for further expression analysis.
In the latter part of this work, the expression level of 8 putative genes encoding ABC- and MFS-transporters – which were selected according to C. rosea IK726 ESTs data – was evaluated on different treatments such as fungal-fungal interaction, zearalenone (ZEA, mycotoxin) and four different fungicides. ‘Delta-delta method’ was adopted as relative quantification method using actin as reference gene.