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Optimization of protein extraction methods for root-associated microbes in maize

ABSTRACT: The goal of this study was to optimize protein extraction methods to study root-associated bacteria in maize. For this we inoculated sterile maize plants with a synthetic community composed of seven different bacteria (Ben Niu et al. PNAS 2017, vol 114, n 12). Then, we extracted proteins from maize roots using eight different protein extraction methods in triplicates. These methods were a combination of different extraction buffers (SDS or Triton-based) and mechanical disruption methods (bead-beating, N2 grinding, glass homogenizer and freeze-thaw cycles). We found that vortexing maize roots with glass beads in PBS yielded the highest numbers of microbial protein identification.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Herbaspirillum Robiniae Chryseobacterium Indologenes Ochrobactrum Pituitosum Enterobacter Ludwigii Pseudomonas Putida Zea Mays (maize) Curtobacterium Pusillum

SUBMITTER: Fernanda Salvato

LAB HEAD: Manuel Kleiner

PROVIDER: PXD026369 | Pride | 2022-09-08

REPOSITORIES: Pride

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Evaluation of Protein Extraction Methods for Metaproteomic Analyses of Root-Associated Microbes.

Salvato Fernanda F Vintila Simina S Finkel Omri M OM Dangl Jeffery L JL Kleiner Manuel M

Molecular plant-microbe interactions : MPMI 20221031 11

Metaproteomics is a powerful tool for the characterization of metabolism, physiology, and functional interactions in microbial communities, including plant-associated microbiota. However, the metaproteomic methods that have been used to study plant-associated microbiota are very laborious and require large amounts of plant tissue, hindering wider application of these methods. We optimized and evaluated different protein extraction methods for metaproteomics of plant-associated microbiota in two ...[more]

PMID: 35876747

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