Quantification of Superoxide and Hydrogen Peroxide in Leaves

Juszczak, Ilona; Baier, Margarete

doi:10.1007/978-1-4939-0844-8_16

Cited by 16 publications

(11 citation statements)

References 18 publications

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“…Histochemical analyses of radical (R • ) and hydrogen peroxide (H 2 O 2 ) levels were performed according to Juszczak and Baier ( 2014 ) with nitroblue tetrazolium [NBT; 1 mg/ml NBT in 10 mM NaN 3 , 8% (m/v) NaCl, 0.2% (w/v) KCl, 1.44%(w/v) Na 2 HPO 4 and 0.24% (w/v) KH 2 PO 4 ; pH 7.4] and 3,3-diaminobenzidine [DAB; 1 mg/ml DAB in 8% (m/v) NaCl, 0.2% (w/v) KCl, 1.44%(w/v) Na 2 HPO 4 and 0.24% (w/v) KH 2 PO 4 ; pH 7.4]. All leaves were transferred immediately into the staining solution upon harvest and infiltrated at very low light intensity with the staining solutions.…”

Section: Methodsmentioning

confidence: 99%

Natural Variation of Cold Deacclimation Correlates with Variation of Cold-Acclimation of the Plastid Antioxidant System in Arabidopsis thaliana Accessions

et al. 2016

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Temperature variations impact on the balance between photosynthetic electron transport and electron-consuming assimilation reactions and transiently increase generation of reactive oxygen species (ROS). Previous studies demonstrated that the expression of C-repeat binding factors (CBFs), which activate cold acclimation reactions, respond to chloroplast ROS signals and that cold deacclimation is partly halted for days after the transfer of acclimated plants to optimal growth conditions in four Arabidopsis accessions from cold-continental habitats. We hypothesized that these accessions differ from others in the regulation of the plastid antioxidant system (PAS). In the present study, we compared the expression intensity of the 12 most prominent PAS genes for peroxidases, superoxide dismutase and low molecular weight antioxidant regenerating enzymes in 10 Arabidopsis accessions with regulation of CBF and COR (cold regulated genes) transcript levels and cold-regulated metabolite levels prior to cold, after 2 week long cold acclimation and during the first 3 days of deacclimation. In the accessions with prolonged activation of cold responses, by trend, weaker induction of various cold-inducible PAS genes and stronger decreases in the expression of negatively cold-regulated PAS genes were observed. Low PAS gene expression delayed the post-cold decrease in H2O2 levels after transfer of the plants from cold to optimal growth conditions. We conclude that weaker expression of various PAS genes in the cold is an adapted strategy of the Arabidopsis accessions N14, N13, Ms-0, and Kas-1 to avoid full inactivation of cold-responses in the first days after the end of the cold period.

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Section: Methodsmentioning

confidence: 99%

Natural Variation of Cold Deacclimation Correlates with Variation of Cold-Acclimation of the Plastid Antioxidant System in Arabidopsis thaliana Accessions

et al. 2016

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“…Nitro blue tetrazolium (NBT) staining for the O 2 level and 3,3´-diaminobenzine (DAB) staining for the H 2 O 2 level were carried out according to Juszczak and Baier (2014) with minor modification. The first or sixth leaves of the plants after 7 d Zn limitation were detached and vacuum-infiltrated with NBT or DAB solutions.…”

Section: Measurements Of O 2 or H 2 O 2 Levelsmentioning

confidence: 99%

Autophagy is induced under Zn limitation and contributes to Zn-limited stress tolerance in Arabidopsis (Arabidopsis thaliana)

Eguchi

Kimura

Makino

et al. 2017

Soil Science and Plant Nutrition

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Autophagy is a degradation system for cellular components conserved in eukaryotes. In Arabidopsis, it is known that autophagy is crucial for growth under dark-induced carbon starvation and N deficiency. However, little is known about the relationship between autophagy and other nutrients. Here, we focused on the relationship between autophagy and Zn nutrition. We found that autophagy-deficient (atg) mutants showed an early senescence phenotype under Zn limitation and limited growth recovery from Zn limitation. Furthermore, we confirmed the induction of autophagy under Zn limitation by expression analysis of autophagy-related genes (ATGs) and imaging analysis of autophagic bodies with green fluorescent protein-ATG8a (GFP-ATG8a). In atg mutants, although the Zn concentrations were similar to those of the wild-type plants, the transcript levels of Zn deficiency-inducible genes fluctuated more, and O 2 and H 2 O 2 levels increased more than in wild-type plants. These results suggest that autophagy is involved in intracellular Zn usage and suppresses the accumulation of reactive oxygen species (ROS) generated by Zn limitation. ARTICLE HISTORY

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“…One senescence down-regulated gene (SDRG), DGR2 , displayed an earlier decrease in expression at day 42 in both cyp79B2/cyp79B3 lines, but an earlier decrease was not observed in a second SDRG, Lhcb2.4 . Accumulation of another marker of leaf senescence, H 2 O 2 , was approximated using DAB staining (Juszczak and Baier, 2014). H 2 O 2 levels were increased in the cyp79B2/cyp79B3 lines at day 49 ( Supplemental Figure 3 ).…”

Section: Resultsmentioning

confidence: 99%

Negative Regulation of Age-Related Developmental Leaf Senescence by the IAOx Pathway, PEN1, and PEN3

et al. 2019

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Early age-related developmental senescence was observed in Arabidopsis cyp79B2/cyp79B3 double mutants that cannot produce indole-3-acetaldoxime (IAOx), the precursor to indole glucosinolates (IGs), camalexin and auxin. The early senescence phenotype was not observed when senescence was induced by darkness. The cyp79B2/cyp79B3 mutants had lower auxin levels, but did not display auxin-deficient phenotypes. Camalexin biosynthesis mutants senesced normally; however, IG transport and exosome-related pen1/pen3 double mutants displayed early senescence. The early senescence in pen1/pen3 mutants depended on salicylic acid and was not observed in pen1 or pen3 single mutants. Quantitation of IGs showed reduced levels in cyp79B2/cyp79B3 mutants, but unchanged levels in pen1/pen3, even though both of these double mutants display early senescence. We discuss how these genetic data provide evidence that IAOx metabolites are playing a protective role in leaf senescence that is dependent on proper trafficking by PEN1 and PEN3, perhaps via the formation of exosomes.

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Quantification of Superoxide and Hydrogen Peroxide in Leaves

Cited by 16 publications

References 18 publications

Natural Variation of Cold Deacclimation Correlates with Variation of Cold-Acclimation of the Plastid Antioxidant System in Arabidopsis thaliana Accessions

Natural Variation of Cold Deacclimation Correlates with Variation of Cold-Acclimation of the Plastid Antioxidant System in Arabidopsis thaliana Accessions

Autophagy is induced under Zn limitation and contributes to Zn-limited stress tolerance in Arabidopsis (Arabidopsis thaliana)

Negative Regulation of Age-Related Developmental Leaf Senescence by the IAOx Pathway, PEN1, and PEN3

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