iTRAQ-based proteomics analysis of autophagy-mediated immune responses against the vascular fungal pathogen Verticillium dahliae in Arabidopsis

Abstract

The mechanisms underlying the functional link between autophagy and plant innate immunity remain largely unknown. In this study, we investigated the autophagy-mediated plant defense responses against Verticillium dahliae (V. dahliae) infection by comparative proteomics and cellular analyses. An assessment of the autophagy activity and disease development showed that autophagic processes were tightly related to the tolerance of Arabidopsis plant to Verticillium wilt. An isobaric tags for relative and absolute quantification (iTRAQ)-based proteomics analysis was performed, and we identified a total of 780 differentially accumulated proteins (DAPs) between wild-type and mutant atg10-1 Arabidopsis plants upon V. dahliae infection, of which, 193 ATG8-family-interacting proteins were identified in silico and their associations with autophagy were verified for several selected proteins. Three important aspects of autophagy-mediated defense against V. dahliae infection were revealed: 1) autophagy is required for the activation of upstream defense responses; 2) autophagy-mediated mitochondrial degradation (mitophagy) occurs and is an important player in the defense process; and 3) autophagy promotes the transdifferentiation of perivascular cells and the formation of xylem hyperplasia, which are crucial for protection against this vascular disease. Together, our results provide several novel insights for understanding the functional association between autophagy and plant immune responses.

Knockout of Pi21 by CRISPR/Cas9 and iTRAQ-Based Proteomic Analysis of Mutants Revealed New Insights into M. oryzae Resistance in Elite Rice Line

“…To perceive the effects of mutations on plant proteome, the proteomics analysis strategy is the most powerful way to explore the diverse changes in complex biological processes and iTRAQ (isobaric tags for relative and absolute quantification)-based proteome analysis is an effective technique for quantitative measurement of protein abundance, regardless of the causes of mutations [ 33 , 34 ]. It has been widely used in plant species to recognize the diverse biological processes including, Arabidopsis [ 35 ], rice [ 36 ], wheat [ 37 ], and maize [ 38 ]. A rising number of CRISPR-Cas9 applications necessitate a good understanding and better management of its efficiency and off-targets.…”

Large Scale Proteomic Data and Network-Based Systems Biology Approaches to Explore the Plant World

“…In addition, only cysteine-containing peptides are analyzed, impairing the quantification of those that do not contain this aminoacid. Many more studies to quantify plant proteins have adopted isobaric tags for relative and absolute quantitation (iTRAQ) [ 108 , 109 , 110 ] and tandem mass tags (TMT) [ 111 ]. In comparison to ICAT, or ICPL [ 112 ], TMT and iTRAQ labeling takes place at the peptide level by means of amine specific reagents which assure a wider peptide labeling without loss of PTM information.…”

TRPM8-regulated calcium mobilization plays a critical role in synergistic chemosensitization of Borneol on Doxorubicin

“…Although these studies suggest the potential application of NB in cancer prevention, it is not clear whether NB can be used to improve the therapeutic efficiency of chemotherapy in lung cancer treatment. RNA-Seq 22 and ITRAQ 23 are widely used to explore the targets of small molecules, proteins, and peptides. However, these strategies are still unable to identify direct interactions between protein targets and drug molecules.…”