Early detection of Botrytis cinerea in strawberry fruit during quiescent infection using selected ion flow tube mass spectrometry (SIFT-MS).

Botrytis cinerea is a devastating pathogen that can cause huge postharvest losses of strawberry. Although this fungus usually infects strawberries through their flowers, symptoms mainly appear when fruit are fully mature. A fast and sensitive method to detect and quantify the fungal infection, prior to symptom development, is, therefore, needed. In this study, we explore the possibility of using the strawberry volatilome to identify biomarkers for B. cinerea infection. Strawberry flowers were inoculated with B. cinerea to mimic the natural infection. First, quantitative polymerase chain reaction (qPCR) was used to quantify B. cinerea in the strawberry fruit. The detection limit of qPCR for B. cinerea DNA extracted from strawberries was 0.01 ng. Subsequently, changes in the fruit volatilome at different fruit developmental stages were characterized using gas chromatography - mass spectrometry (GC-MS) and selected ion flow tube mass spectrometry (SIFT-MS). Based on GC-MS data, 1-octen-3-ol produced by B. cinerea was confirmed as a potential biomarker of B. cinerea infection. Moreover, the product ion NO 127, obtained by SIFT-MS measurements, was proposed as a potential biomarker for B. cinerea infection by comparing its relative level with that of 1-octen-3-ol (obtained by GC-MS) and B. cinerea (obtained by qPCR). Separate PLS regressions were carried out for each developmental stages, and 11 product ions were significantly altered at all developmental stages. Finally, PLS regressions using these 11 ions as variables allowed the discrimination between samples containing different amount of B. cinerea. This work showed that profiling the fruit's volatilome using SIFT-MS can be used as a potential alternative to detect B. cinerea during the quiescent stage of B. cinerea infection prior to symptom development. Moreover, the corresponding compounds of potential biomarkers suggest that the volatile changes caused by B. cinerea infection may contribute to strawberry defense.