Detection of Hydroxyl Radical in Intact Cells of Chlorella Vulgaris

Hirayama, Shin; Ueda, Ryohei; Sugata, Kiyoshi

doi:10.3109/10715769509064019

Cited by 13 publications

(9 citation statements)

References 7 publications

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“…Although 1 O 2 is probably the primary ROS product, other ROS have also been identified in photosynthetic tissues under PI. These include various oxygen-and carbon-centred radicals [23,[38][39][40][41][42][43]. Using TEMPO-9-AC as a broad-range radical probe, we attempted to trap radicals in PIexposed leaves.…”

Section: Response To 1 O 2 From Photoinhibitionmentioning

confidence: 99%

A comparative study of fluorescent singlet oxygen probes in plant leaves

Hideg

2008

Open Life Sciences

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Four fluorescent singlet oxygen sensors: DanePy, its oxalate salt, Singlet Oxygen Sensor Green and MVP, were infiltrated into tobacco leaves and tested for toxicity, subcellular localization, light sensitivity and capacity to trap the singlet oxygen produced in photoinhibition. For reference, a broad sensitivity free radical probe, TEMPO-9-AC, was also included. Photochemical yield was approximately 15% and 10% inhibited by Singlet Oxygen Sensor Green and MVP, respectively, but was not significantly affected by the other probes. Under photoinhibitory conditions, brought about by irradiating lincomycin-treated leaves with strong photosynthetically active radiation, DanePy and Singlet Oxygen Sensor Green were responsive. Singlet Oxygen Sensor Green was also reactive to low, non-photoinhibitory light exposure of the leaf, which was not characteristic to the other probes. MVP did not respond to singlet oxygen which can partly be explained by a possible attenuation of its blue emission in the leaf, as shown by the example TEMPO-9-AC. DanePy-oxalate did not respond to photosynthetic singlet oxygen due to lack of its penetration into photosynthetic tissue and hence could be useful in detecting any singlet oxygen which escapes from a chloroplast initiation site. DanePy was localized in the chloroplasts, while Singlet Oxygen Sensor Green was mainly found in the epidermal cells preferentially associated with the nucleus.

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Section: Response To 1 O 2 From Photoinhibitionmentioning

confidence: 99%

A comparative study of fluorescent singlet oxygen probes in plant leaves

Hideg

2008

Open Life Sciences

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“…Spin trapping EPR spectroscopy proved useful in detecting ROS in isolated photosynthetic membranes exposed to strong light (Hideg et al 1994a, Hideg et al 1995, Hirayama et al 1995, Yruela et al 1996, or UV radiation (Hideg and Vass 1996). However, its application to intact leaves was limited due to the reduction of spin adducts to the hydroxylamine form by leaf metabolites (Hideg et al 1994b) and to technical problems caused by the high water content of leaves (Hideg et al 2000a).…”

Section: Introductionmentioning

confidence: 99%

Detection of Singlet Oxygen and Superoxide with Fluorescent Sensors in Leaves Under Stress by Photoinhibition or UV Radiation

Hideg

Barta

Kálai

et al. 2002

Plant and Cell Physiology

215

163

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;In order to understand the physiological functions of reactive oxygen species (ROS) generated in leaves, their direct measurement in vivo is of special importance. Here we report experiments with two dansyl-based ROS sensors, the singlet oxygen specific DanePy and HO-1889NH, which is reactive to both singlet oxygen and superoxide radicals. Here we report in vivo detection of 1 O 2 and O 2 -• by fluorescence quenching of two dansyl-based ROS sensors, the 1 O 2 specific DanePy and HO-1889NH, which was reactive with both 1 O 2 and O 2 -• . The ROS sensors were administered to spinach leaves through a pinhole, and then the leaves were exposed to either excess photosynthetically active radiation or UV (280-360 nm) radiation. Microlocalization of the sensors' fluorescence and its ROS-induced quenching was followed with confocal laser scanning microscopy and with fluorescence imaging. These sensors were specifically localized in chloroplasts. Quenching analysis indicated that the leaves exposed to strong light produced 1 O 2 , but hardly any O 2 -• . On the other hand, the dominant ROS in UV-irradiated leaves was O 2 -• , while 1 O 2 was minor.

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“…The procedure followed was similar to that followed by Hirayama et al (1995) on C. vulgaris. The trapping was done with intact cells of Spirulina.…”

Section: Esr Measurementsmentioning

confidence: 99%

“…Hydroxyl radicals have been shown to be involved during photoinhibition and accumulation of oxygen in Chlorella vulgaris (Hirayama et al, 1995(Hirayama et al, , 1996. Also Nishiyama et al (2001) have attributed the hydroxyl radical to be the main ROS involved in causing damage in cyanobacterium Synechocystis sp.…”

Section: Introductionmentioning

confidence: 99%

Responses of the photosynthetic machinery of Spirulina maxima to induced reactive oxygen species

Ganesh

Manoharan

Suraishkumar

2006

Biotech & Bioengineering

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The photosynthetic machinery of Spirulina maxima was studied when subjected to induced reactive oxygen species (ROS) to examine the organism's responses to stress. Significant decreases in both photosynthetic efficiency and growth rate were observed. Exposure to 0.01 mmol H(2)O(2)/(g cell), which induced the lowest specific intracellular ROS level (siROS) led to a 15% decrease in specific growth rate; an increase in siROS by 70-fold led to a 25% decrease in specific growth rate. Similarly, siROS induced by 0.01 mmol H(2)O(2)/(g cell) led to 15% inhibition in photosynthetic efficiency, while an increase in siROS by 40- or 70-fold led to about 60% inhibition in photosynthetic efficiency. To further understand the effects of induced ROS on photosynthetic machinery, we performed a detailed pigmentation analysis as well as analyzed Phycobilisomes (PBS), Photosystem II (PSII), and Photosystem I (PSI), the three important components of cyanobacterial photosynthetic apparatus. We found carotenoids (beta-carotene and lutein) to be most sensitive to siROS. Also, specific levels of phycocyanin and allophycocyanin, which are important PBS pigments, decreased significantly in response to H(2)O(2). Further, electron transport assays revealed that ROS cause damage primarily to PSII, whereas they do not significantly affect PSI in comparison; siROS induced by 0.01 mmol H(2)O(2)/(g cell) led to a 15% inhibition of PSII, and increase in siROS by 9-, 40-, and 70-fold led to 22%, 36%, and 46% inhibition, respectively.

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Detection of Hydroxyl Radical in Intact Cells of Chlorella Vulgaris

Cited by 13 publications

References 7 publications

A comparative study of fluorescent singlet oxygen probes in plant leaves

A comparative study of fluorescent singlet oxygen probes in plant leaves

Detection of Singlet Oxygen and Superoxide with Fluorescent Sensors in Leaves Under Stress by Photoinhibition or UV Radiation

Responses of the photosynthetic machinery of Spirulina maxima to induced reactive oxygen species

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