Direct identification of Monilinia brown rot fungi on infected fruits by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry
Open access
Date
2016Type
- Journal Article
ETH Bibliography
yes
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Abstract
Background
Brown rot of stone and pome fruit is a serious fungal disease that is mainly caused by four species in the genus Monilinia. Of these four species, Monilinia fructicola is the most devastating pathogen and of particular concern because it undergoes sexual recombination and has recently been introduced to Europe. So far, Monilinia diagnosis required a multiplex PCR analysis and gel electrophoresis. In contrast, intact-protein biotyping by mass spectrometry is considerably faster and cheaper. However, it usually requires an in vitro cultivation step prior to the MALDI analysis. It was thus attempted to establish a method for the identification of Monilinia species by MALDI biotyping with fungal material derived directly from infected fruits; without an in vitro cultivation step.
Results
To simplify and render MALDI biotyping of fungi more reliable, an improved protocol for the preparation of crude protein extracts and for collecting MALDI-TOF MS data for biotyping was developed. We generated reference spectra for all four Monilinia brown rot fungi and were able to reliably identify Monilinia species based on fungal material that was collected directly from infected fruits. This method allowed the correct, fast and economic identification of M. fructicola and M. laxa, while M. fructigena and M. polystroma could not be distinguished reliably.
Conclusions
MALDI biotyping may be used as an economical tool for the routine diagnosis of Monilinia brown rot fungi on infected fruits. Show more
Permanent link
https://doi.org/10.3929/ethz-b-000318208Publication status
publishedExternal links
Journal / series
Chemical and Biological Technologies in AgricultureVolume
Pages / Article No.
Publisher
SpringerSubject
Biotyping; Intact‑protein profiling; MALDI‑TOF/TOF MS; MoniliniaOrganisational unit
02207 - Functional Genomics Center Zurich / Functional Genomics Center Zurich
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ETH Bibliography
yes
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