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Direct identification of Monilinia brown rot fungi on infected fruits by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry


Freimoser, Florian Matthias; Hilber-Bodmer, Maja; Brunisholz, René; Drissner, David (2016). Direct identification of Monilinia brown rot fungi on infected fruits by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. Chemical and Biological Technologies in Agriculture, 3:7.

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.

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.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Functional Genomics Center Zurich
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Language:English
Date:2016
Deposited On:18 Jan 2017 16:34
Last Modified:28 Apr 2017 06:48
Publisher:SpringerOpen
ISSN:2196-5641
Publisher DOI:https://doi.org/10.1186/s40538-016-0058-4

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