Hydrogen Peroxide Measurement in Arabidopsis Root Tissue Using Amplex Red

Brumbarova, Tzvetina; Le, Cham Thi Tuyet; Bauer, Petra

doi:10.21769/bioprotoc.1999

Cited by 14 publications

(8 citation statements)

References 4 publications

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“…Firstly, the production of H 2 O 2 was monitored by UV-vis spectroscopy due to its ability to oxidize Amplex Red to the coloured Resorufin (l abs = 570 nm) in the presence of horseradish peroxidase (HRP). 65,66 As depicted in Fig. 5b, the presence of FD 3 K and FE 3 K induces a strong decrease of H 2 O 2 formation by 88% and 89%, corresponding to 2.0 mM and 1.8 mM H 2 O 2 , respectively, compared to the negative control EDTA (16.7 mM H 2 O 2 formed).…”

Section: Antioxidant Propertiesmentioning

confidence: 88%

Both metal-chelating and free radical-scavenging synthetic pentapeptides as efficient inhibitors of reactive oxygen species generation

et al. 2020

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Section: Antioxidant Propertiesmentioning

confidence: 88%

Both metal-chelating and free radical-scavenging synthetic pentapeptides as efficient inhibitors of reactive oxygen species generation

et al. 2020

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“…The frozen leaves were ground, and 5-10 mg of samples were extracted with 1 ml of 20-mM K 2 HPO 4 (pH 6.5), homogenized for 5 min at 4 C, and then centrifuged for 3 min at 16,200 g at 4 C. The H 2 O 2 detection assay by Amplex Red (Thermo Fisher Scientific, Waltham, MA,USA) was performed on the supernatant, as described in Brumbarova, Le, and Bauer (2016) with some modifications, that is, the reaction was incubated for 15 min instead of 30 min. Samples are combusted at 1,000 C in a reactor packed with chromium oxide and silvered copper oxide.…”

Section: H 2 O 2 Quantificationmentioning

confidence: 99%

Silencing of OsCV (chloroplast vesiculation) maintained photorespiration and N assimilation in rice plants grown under elevated CO₂

Umnajkitikorn

Sade

Wilhelmi

et al. 2020

Plant Cell & Environment

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High CO2 concentrations stimulate net photosynthesis by increasing CO2 substrate availability for Rubisco, simultaneously suppressing photorespiration. Previously, we reported that silencing the chloroplast vesiculation (cv) gene in rice increased source fitness, through the maintenance of chloroplast stability and the expression of photorespiration‐associated genes. Because high atmospheric CO2 conditions diminished photorespiration, we tested whether CV silencing might be a viable strategy to improve the effects of high CO2 on grain yield and N assimilation in rice. Under elevated CO2, OsCV expression was induced, and OsCV was targeted to peroxisomes where it facilitated the removal of OsPEX11‐1 from the peroxisome and delivered it to the vacuole for degradation. This process correlated well with the reduction in the number of peroxisomes, the decreased catalase activity and the increased H2O2 content in wild‐type plants under elevated CO2. At elevated CO2, CV‐silenced rice plants maintained peroxisome proliferation and photorespiration and displayed higher N assimilation than wild‐type plants. This was supported by higher activity of enzymes involved in NO3− and NH4+ assimilation and higher total and seed protein contents. Co‐immunoprecipitation of OsCV‐interacting proteins suggested that, similar to its role in chloroplast protein turnover, OsCV acted as a scaffold, binding peroxisomal proteins.

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“…H 2 O 2 in root samples was measured using the Amplex Red H 2 O 2 –peroxidase assay kit (Molecular Probes) as described in detail in Brumbarova et al (2016 a ). The H 2 O 2 content values for three independent replicates per condition were measured, averaged, and the SD was calculated.…”

Section: Methodsmentioning

confidence: 99%

Reactive oxygen species coordinate the transcriptional responses to iron availability in Arabidopsis

Mark

Ivanov

Eutebach

et al. 2020

Journal of Experimental Botany

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Reactive oxygen species play a central role in the regulation of plant responses to environmental stress. Under prolonged iron (Fe) deficiency, increased levels of hydrogen peroxide (H2O2) initiate signaling events, resulting in the attenuation of Fe acquisition through the inhibition of FER-LIKE IRON DEFICIENCY-INDUCED TRANSCRIPTION FACTOR (FIT). As this H2O2 increase occurs in a FIT-dependent manner, our aim was to understand the processes involved in maintaining H2O2 levels under prolonged Fe deficiency and the role of FIT in this process. We identified CAT2 gene, encoding one of the three Arabidopsis catalase isoforms, as regulated by FIT. CAT2 loss-of-function plants displayed severe susceptibility to Fe deficiency and greatly increased H2O2 levels in roots. Analysis of the Fe homeostasis transcriptional cascade revealed that H2O2 influences the gene expression of downstream regulators FIT, BHLHs of group Ib and POPEYE (PYE), however H2O2 did not affect their upstream regulators, such as BHLH104 and ILR3. Our data shows that FIT and CAT2 participate in a regulatory loop between H2O2 and prolonged Fe deficiency.

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Hydrogen Peroxide Measurement in Arabidopsis Root Tissue Using Amplex Red

Cited by 14 publications

References 4 publications

Both metal-chelating and free radical-scavenging synthetic pentapeptides as efficient inhibitors of reactive oxygen species generation

Both metal-chelating and free radical-scavenging synthetic pentapeptides as efficient inhibitors of reactive oxygen species generation

Silencing of OsCV (chloroplast vesiculation) maintained photorespiration and N assimilation in rice plants grown under elevated CO₂

Reactive oxygen species coordinate the transcriptional responses to iron availability in Arabidopsis

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Hydrogen Peroxide Measurement in Arabidopsis Root Tissue Using Amplex Red

Cited by 14 publications

References 4 publications

Both metal-chelating and free radical-scavenging synthetic pentapeptides as efficient inhibitors of reactive oxygen species generation

Both metal-chelating and free radical-scavenging synthetic pentapeptides as efficient inhibitors of reactive oxygen species generation

Silencing of OsCV (chloroplast vesiculation) maintained photorespiration and N assimilation in rice plants grown under elevated CO2

Reactive oxygen species coordinate the transcriptional responses to iron availability in Arabidopsis

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Silencing of OsCV (chloroplast vesiculation) maintained photorespiration and N assimilation in rice plants grown under elevated CO₂