Premium
Rapid glycosyl‐inositol‐phospho‐ceramide fingerprint from filamentous fungal pathogens using the MALDI Biotyper Sirius system
Author(s) -
Saromi Kofo,
England Philippa,
Tang Wenhao,
Kostrzewa Markus,
Corran Andy,
Woscholski Rudiger,
LarrouyMaumus Gerald
Publication year - 2020
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.8904
Subject(s) - chemistry , neurospora crassa , chromatography , mass spectrometry , matrix assisted laser desorption/ionization , biochemistry , desorption , organic chemistry , adsorption , mutant , gene
Rationale Glycosyl‐inositol‐phospho‐ceramides (GIPCs) or glycosylphosphatidylinositol‐anchored fungal polysaccharides are known to be major lipids in plant and fungal plasma membranes and to play an important role in stress adaption. However, their analysis remains challenging due to the several steps involved for their extractions and purifications prior to mass spectrometric analysis. To address this challenge, we developed a rapid and sensitive method to identify GIPCs from the four common fungal plant pathogens Botrytis cinerea , Fusarium graminearium , Neurospora crassa and Ustilago maydis . Methods Fungal plant pathogens were cultured, harvested, heat‐inactivated and washed three times with double‐distilled water. Intact fungi were deposited on a matrix‐assisted laser desorption ionization (MALDI) target plate, mixed with the matrix consisting of a 9:1 mixture of 2,5‐dihydroxybenzoic acid and 2‐hydroxy‐5‐methoxybenzoic acid solubilized at 10 mg/mL in chloroform–methanol (9:1 v/v) and analyzed using a Bruker MALDI Biotyper Sirius system in the linear negative ion mode. Mass spectra were acquired from m / z 700 to 2000. Results MALDI time‐of‐flight (TOF) mass spectrometric analysis of cultured fungi showed clear signature of GIPCs in B. cinerea , F. graminearium , N. crassa and U. maydis .Conclusions We have demonstrated that routine MALDI‐TOF in the linear negative ion mode combined with an apolar solvent system to solubilize the matrix is applicable to the detection of filamentous fungal GIPCs.