Matrix assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry for identification of plant pathogenic Alternaria species

Chowdappa, P.; Lakshmi, M. Jyothi; Madhura, S.

doi:10.1007/s12600-012-0276-7

Cited by 12 publications

(5 citation statements)

References 36 publications

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“…Studies of filamentous fungi by MALDI-TOF MS based on vegetative cells usually yielded satisfactory identifications for species of Trichoderma (de Respinis et al 2010); these authors concluded that MALDI-TOF MS analyses of 2 d old mycelial samples were useful for species delimitation in this genus. Similarly Chowdappa et al (2013) showed significant MALDI-TOF capacity to identify Alternaria species by analyzing intact 5 d old mycelium. However, we observed both higher quality and better reproducibility for protein spectra using only conidia of Metarhizium.…”

Section: Discussionmentioning

confidence: 99%

MALDI-TOF mass spectrometry applied to identifying species of insect-pathogenic fungi from theMetarhizium anisopliaecomplex

et al. 2014

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Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has proven to be a powerful tool for taxonomic resolution of microorganisms. In this proof-of-concept study, we assessed the effectiveness of this technique to track the current gene sequence-based phylogenetic classification of species in the Metarhizium anisopliae complex. Initially the phylogenetic analysis of 5' strains by sequencing of the 59' end of the TEF-1α gene region revealed seven species within M. anisopliae sensu lato and two varieties outside this complex. Because initial studies on MS profiles from different cell types showed that mycelial fragments or conidia produced on nutrient-poor medium may yield too much background noise, all subsequent spectrometric analyses were performed with acidhydrolyzed conidia from 10-12 d old PDA cultures. The initial MALDI-TOF reference library included protein spectral profiles from nine taxonomically distinct, molecularly identified isolates sharing high genetic homology with the ex-type or ex-epitype isolates of these taxa in Metarhizium. A second reference library added one isolate each for M. anisopliae sensu stricto and M. robertsii. The second, larger reference library (including 11 taxa) allowed nearly perfect MALDI-TOF matching of DNA-based species identification for the 40 remaining isolates molecularly recognized as M. anisopliae sensu stricto (n = 19), M. robertsii (n = 6), M. majus (n = 3), M. lepidiotae (n = 1), M. acridum (n = 3), M. flavoviride var. pemphigi (n = 1), plus seven unidentified strains (six of them phylogenetically close to M. anisopliae sensu stricto and one outside the Metarhizium pingshaense-anisopliae-robertsii-brunneum clade). Due to the increasing frequency of phylogenetically (genomically) based taxonomic revisions of fungi, this approach is especially useful for culture collections, because once the protein profiles of Metarhizium isolates are obtained taxonomic updating of MALDI-TOF library data is easily accomplished by comparing stored profiles with those of newly proposed taxa.

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Section: Discussionmentioning

confidence: 99%

MALDI-TOF mass spectrometry applied to identifying species of insect-pathogenic fungi from theMetarhizium anisopliaecomplex

et al. 2014

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“…Further, Chowdappa et al , 112 have distinguished plant pathogenic fungi, based on protein markers expressed in intact cells using MALDI MS. The protein fingerprint-based analysis grouped 12 Alternaria species (with high similarity in spectral characteristics) out of 60 isolates of Alternaria species studied, using direct intact MALDI MS.…”

Section: Maldi Ms-based Identification Of Plant-associated Microbesmentioning

confidence: 99%

A systematic assessment of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) application for rapid identification of pathogenic microbes that affect food crops: delivered and future deliverables

et al. 2023

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“…Metarhizium samples were inoculated on PDA and SDA medium (Lopes et al, 2014). De Respins et al (2010) used 2 days old Trichoderma samples and Chowdappa et al (2013) incubated Alternaria for 5 days. The optimal technique for Beauveria species has not been tested before.…”

Section: Spectrum Analysis Of Proteins By Maldi-tof Mass Spectrometrymentioning

confidence: 99%

Use of MALDI-TOF mass spectrometry in rapid identification of Beauveria bassiana and Beauveria pseudobassiana

Hricáková¹,

Medo²,

Hleba³

et al. 2018

JCEA

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Species of the genus Beauveria are entomopathogenic fungi used in biological control to protect against insect pests. Identification of species is based on observation of morphological, phylogenetic and genetic differences. Currently, MALDI-TOF (Matrix Assisted Laser Desorption/Ionization-Time of Flight) mass spectrometry is often used in microbial diagnostic but identification of Beauveria species has not been tested before. The aim of this work was the possibilities of this method and to compare it with genetic analysis. Twenty strains of B. bassiana and B. pseudobassiana isolated from insect cadavers, mycoinsecticides and soil samples were used. Sequences of ITS (Internal Transcribed Spacer) region were used for genetic identification of strains. Sequences and results from protein analysis of isolates were compared with the reference strains of B. bassiana and B. pseudobassiana. Results of MALDI Biotyper softvare identification were correct for all strains. Identification was reliable with high log score (>2.1) for most of strains. Score value less than 1 was observed only in a single case -strain GHA isolated from mycoinsecticide Botanigard.The result of both analyses was also dendrogram (phylogenetic tree) which showed high degree of result consistence. MALDI-TOF identification of fungi from Beauveria genus is usable for fast and reliable identification of entomopathogenic fungi isolates acquired from environment and results can be utilized in conservation biocontrol strategies in agriculture and forest ecosystems. Keywords:Beauveria, genetic analysis, spectrum analysis of proteins

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Matrix assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry for identification of plant pathogenic Alternaria species

Cited by 12 publications

References 36 publications

MALDI-TOF mass spectrometry applied to identifying species of insect-pathogenic fungi from theMetarhizium anisopliaecomplex

MALDI-TOF mass spectrometry applied to identifying species of insect-pathogenic fungi from theMetarhizium anisopliaecomplex

A systematic assessment of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) application for rapid identification of pathogenic microbes that affect food crops: delivered and future deliverables

Use of MALDI-TOF mass spectrometry in rapid identification of Beauveria bassiana and Beauveria pseudobassiana

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