Organic radical imaging in plants: Focus on protein radicals

Kumar, Aditya; Prasad, Ankush; Sedlářová, Michaela; Pospı́šil, Pavel

doi:10.1016/j.freeradbiomed.2018.10.428

Cited by 10 publications

(12 citation statements)

References 33 publications

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“…In photosynthetic organism, generation of reactive oxygen species (ROS) is a quite universal and fast defense mechanism, known to be associated with various stresses both in vivo and in vitro (Yadav and Pospíšil, 2012;Prasad et al, 2015;Prasad et al, 2016;Prasad et al, 2017a;Prasad et al, 2018;Kumar et al, 2019), in both local as well as systemic responses (Grant and Loake, 2000;Slesak et al, 2007). In tomato, it has been observed that hydrogen peroxide (H 2 O 2 ) was produced at the site within an hour of wounding and its level was enhanced even in distant part (upper unwounded leaves) in the following 4 to 6 h (Orozco-Cardenas and Ryan, 1999).…”

Section: Introductionmentioning

confidence: 99%

Reactive Oxygen Species as a Response to Wounding: In Vivo Imaging in Arabidopsis thaliana

et al. 2020

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Mechanical injury or wounding in plants can be attributed to abiotic or/and biotic causes. Subsequent defense responses are either local, i.e. within or in the close vicinity of affected tissue, or systemic, i.e. at distant plant organs. Stress stimuli activate a plethora of early and late reactions, from electric signals induced within seconds upon injury, oxidative burst within minutes, and slightly slower changes in hormone levels or expression of defense-related genes, to later cell wall reinforcement by polysaccharides deposition, or accumulation of proteinase inhibitors and hydrolytic enzymes. In the current study, we focused on the production of reactive oxygen species (ROS) in wounded Arabidopsis leaves. Based on fluorescence imaging, we provide experimental evidence that ROS [superoxide anion radical (O 2 •−) and singlet oxygen (1 O 2)] are produced following wounding. As a consequence, oxidation of biomolecules is induced, predominantly of polyunsaturated fatty acid, which leads to the formation of reactive intermediate products and electronically excited species.

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Section: Introductionmentioning

confidence: 99%

Reactive Oxygen Species as a Response to Wounding: In Vivo Imaging in Arabidopsis thaliana

et al. 2020

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“…Formation of protein radicals in leaves and chloroplasts was visualized by confocal laser scanning microscopy (Kumar et al, 2019). Sliced leaf pieces and chloroplasts were incubated either in dark or red light (1000 μmol photons m –2 s –1 ) for 30 min at room temperature, in the presence of MES-NaOH buffer (40 mM, pH 6.5), spin trap DMPO (50 mM), anti-DMPO antibody (5 μg ml –1 ) conjugated with fluorescein isothiocyanate (FITC) and Triton X-100 (0.001%).…”

Section: Methodsmentioning

confidence: 99%

“…In this technique, the reaction of spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) with an organic (proteins, lipids, and nucleic acids) radical forms a stable nitrone adduct (DMPO-P adduct), which is identified by an anti-DMPO antibody raised against nitrone moiety of the adduct (Mason, 2016). Recently, protein radicals have been visualized for the first time in Arabidopsis plants using the immuno-spin trapping technique (Kumar et al, 2019). The authors showed by confocal laser scanning microscopy using fluorescein conjugated with anti-DMPO antibody that protein radicals are formed predominantly in the chloroplasts located at the periphery of the cells and distributed uniformly throughout the grana stack.…”

Section: Introductionmentioning

confidence: 99%

Characterization of Protein Radicals in Arabidopsis

et al. 2019

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Oxidative modification of proteins in photosystem II (PSII) exposed to high light has been studied for a few decades, but the characterization of protein radicals formed by protein oxidation is largely unknown. Protein oxidation is induced by the direct reaction of proteins with reactive oxygen species known to form highly reactive protein radicals comprising carbon-centered (alkyl) and oxygen-centered (peroxyl and alkoxyl) radicals. In this study, protein radicals were monitored in Arabidopsis exposed to high light by immuno-spin trapping technique based on the detection of 5,5-dimethyl-1-pyrroline N-oxide (DMPO) nitrone adducts using the anti-DMPO antibody. Protein radicals were imaged in Arabidopsis leaves and chloroplasts by confocal laser scanning microscopy using fluorescein conjugated with the anti-DMPO antibody. Characterization of protein radicals by standard blotting techniques using PSII protein specific antibodies shows that protein radicals are formed on D1, D2, CP43, CP47, and Lhcb3 proteins. Protein oxidation reflected by the appearance/disappearance of the protein bands reveals that formation of protein radicals was associated with protein fragmentation (cleavage of the D1 peptide bonds) and aggregation (cross-linking with another PSII subunits). Characterization of protein radical formation is important for better understating of the mechanism of oxidative modification of PSII proteins under high light.

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“…Data presented herein bring together microscopic techniques used to visualize the formation of ROS (HO • and 1 O 2 ) within Arabidopsis leaves in the first stage and detection of organic radical formation in the second stage of stress-induced protein oxidation as suggested in [1] . Plants were subjected to high white light stress (1500 µmol photons m −2 s −1 ) at low temperature (8 °C) for 13 h, i.e.…”

Section: Datamentioning

confidence: 99%

“… Data source location Department of Biophysics, Centre of the Region Haná for Biotechnological and Agricultural Research and Department of Botany, Palacký University, Olomouc, Czech Republic. Loc: 49°34′33.828″N, 17°16′54.658″E Data accessibility The data are available within this article Related research article Aditya Kumar, Ankush Prasad, Michaela Sedlářová, Pavel Pospíšil, Organic radical imaging in plants: Focus on protein radicals (in press, Free Radical Biology and Medicine) doi.org/10.1016/j.freeradbiomed.2018.10.428 [1] . …”

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confidence: 99%

Data on detection of singlet oxygen, hydroxyl radical and organic radical in Arabidopsis thaliana

et al. 2018

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This article contains data related to the research article entitled, “Organic radical imaging in plants: Focus on protein radicals” (Kumar et al., 2018). The data presented herein focus on reactive oxygen species (ROS) and organic radical formed within photosynthetic tissues of Arabidopsis thaliana during high light stress and includes (1) Confocal laser scanning microscopic images using 3′-p-(hydroxyphenyl) fluorescein (HPF) as specific probe for the detection of hydroxyl radical (HO•); (2) Confocal laser scanning microscopic images using Singlet Oxygen Sensor Green (SOSG) as a specific probe for the detection of singlet oxygen (1O2) and; (3) Electron paramagnetic resonance (EPR) spectroscopy using spin traps for the detection of organic radical.

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Organic radical imaging in plants: Focus on protein radicals

Cited by 10 publications

References 33 publications

Reactive Oxygen Species as a Response to Wounding: In Vivo Imaging in Arabidopsis thaliana

Reactive Oxygen Species as a Response to Wounding: In Vivo Imaging in Arabidopsis thaliana

Characterization of Protein Radicals in Arabidopsis

Data on detection of singlet oxygen, hydroxyl radical and organic radical in Arabidopsis thaliana

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