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Plant function and evolutionary biology
RESEARCH ARTICLE

Effect of hydrogen peroxide on catalase gene expression, isoform activities and levels in leaves of potato sprayed with homobrassinolide and ultrastructural changes in mesophyll cells

José M. Almeida A B , Fernanda Fidalgo A B , Ana Confraria A B , Arlete Santos A B , Helena Pires A and Isabel Santos A B C
+ Author Affiliations
- Author Affiliations

A Institute for Molecular and Cellular Biology, University of Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal.

B Botany Department, School of Sciences, University of Porto, Rua do Campo Alegre, 1191, 4150-180 Porto, Portugal.

C Corresponding author. Email: isantos@ibmc.up.pt

Functional Plant Biology 32(8) 707-720 https://doi.org/10.1071/FP04235
Submitted: 11 December 2004  Accepted: 6 May 2005   Published: 3 August 2005

Abstract

The effect of hydrogen peroxide (H2O2) on catalase (CAT) isoform activities and amounts and on mRNA levels was studied in leaves from potato plants untreated and treated with homobrassinolide (HBR). Northern blot analysis revealed that 100 mm H2O2 supplied through the leaf petiole for 4 h did not induce CAT expression. In contrast, CAT1 and CAT2 responded differently to longer treatment, as CAT2 transcript levels increased markedly whereas CAT1 transcript levels remained unchanged. Western blot analysis showed disparity between the level of CAT1 transcript and CAT1 amount, which actually decreased after 28 h. CAT2 amount correlated well with transcript accumulation and CAT2 activity as visualised by zymogram analysis. H2O2 modified the relative importance of CAT isoforms. After 4 h, CAT1 was prevalent in untreated and H2O2-treated leaves. After 28 h, CAT2 was prevalent in H2O2-treated leaves; therefore, the quantified increase in total CAT activity in these leaves was due to the rise in CAT2. HBR pre-treatment increased CAT2 basal level not changing the pattern of CAT responses to H2O2, only lowering its amplitude. Even so, ultrastructural studies showed that HBR significantly reduced H2O2 negative effects on cellular sub-structures, allowing better recovery of affected structures and reducing the macroscopic injury symptoms on leaves, thus data point to a HBR protective role.

Keywords: brassinosteroids, catalase gene expression, catalase isozymes, hydrogen peroxide, oxidative stress.


Acknowledgments

The work was supported by the Fundação para a Ciência e Tecnologia (FCT, Lisboa, Portugal — project POCTI / BME / 33044 / 2000).


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