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Publication details

Document type
Journal articles

Document subtype
Full paper

Title
An Optimized in situ Quantification Method of Leaf H2O2 Unveils Interaction Dynamics of Pathogenic and Beneficial Bacteria in Wheat

Participants in the publication
Pablo Carril (Author)
Anabela Bernardes da Silva (Author)
Dep. Biologia Vegetal
 BioISI
 Rogério Tenreiro (Author)
Cristina Cruz (Author)
Dep. Biologia Vegetal
 cE3c
Summary
Hydrogen peroxide (H2O2) functions as an important signaling molecule in plants during biotic interactions. However, the extent to which H2O2 accumulates during these interactions and its implications in the development of disease symptoms is unclear. In this work, we provide a step-by-step optimized protocol for in situ quantification of relative H2O2 concentrations in wheat leaves infected with the pathogenic bacterium Pseudomonas syringae pv. atrofaciens (Psa), either alone or in the presence of the beneficial bacterium Herbaspirillum seropedicae (RAM10). This protocol involved the use of 3-3′diaminobenzidine (DAB) staining method combined with image processing to conduct deconvolution and downstream analysis of the digitalized leaf image. The application of a linear regression model allowed to relate the intensity of the pixels resulting from DAB staining with a given concentration of H2O2. Decreasing H2O2 accumulation patterns were detected at increasing distances from the site of pathogen infection, and H2O2 concentrations were different depending on the bacterial combinations tested. Notably, Psa-challenged plants in presence of RAM10 accumulated less H2O2 in the leaf and showed reduced necrotic symptoms, pointing to a potential role of RAM10 in reducing pathogen-triggered H2O2 levels in young wheat plants.

Date of Publication
2020-06-23

Institution
FACULDADE DE CIÊNCIAS DA UNIVERSIDADE DE LISBOA

Where published
Frontiers in Plant Science

Publication Identifiers
ISSN - 1664-462X

Publisher
Frontiers Media SA

Volume
11

Document Identifiers
DOI - https://doi.org/10.3389/fpls.2020.00889
URL - http://dx.doi.org/10.3389/fpls.2020.00889

Rankings
Web Of Science Q1 (2019) - 4.402 - PLANT SCIENCES - SCIE
SCOPUS Q1 (2019) - 7.8 - Plant Science

Keywords
hydrogen peroxide (H2O2) biotic interactions image processing color deconvolution 3- 3 0diaminobenzidine (DAB)

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APA
Pablo Carril, Anabela Bernardes da Silva, Rogério Tenreiro, Cristina Cruz, (2020). An Optimized in situ Quantification Method of Leaf H2O2 Unveils Interaction Dynamics of Pathogenic and Beneficial Bacteria in Wheat. Frontiers in Plant Science, 11, ISSN 1664-462X. eISSN . http://dx.doi.org/10.3389/fpls.2020.00889

IEEE
Pablo Carril, Anabela Bernardes da Silva, Rogério Tenreiro, Cristina Cruz, "An Optimized in situ Quantification Method of Leaf H2O2 Unveils Interaction Dynamics of Pathogenic and Beneficial Bacteria in Wheat" in Frontiers in Plant Science, vol. 11, 2020. 10.3389/fpls.2020.00889

BIBTEX
@article{47757, author = {Pablo Carril and Anabela Bernardes da Silva and Rogério Tenreiro and Cristina Cruz}, title = {An Optimized in situ Quantification Method of Leaf H2O2 Unveils Interaction Dynamics of Pathogenic and Beneficial Bacteria in Wheat}, journal = {Frontiers in Plant Science}, year = 2020, volume = 11 }