Microplate quantification of enzymes of the plant ascorbate–glutathione cycle

Murshed, Ramzi; Lopez-Lauri, Félicie; Sallanon, Huguette

doi:10.1016/j.ab.2008.07.020

Cited by 244 publications

(132 citation statements)

References 9 publications

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“…The reaction mixture contained 25 mM K-phosphate buffer (pH 7.8), 5 mM GSSG, 1.2 mM NADPHÁNa 4 and 50 lg protein (200 ll final volume; 96-well microplates). The reaction was measured every 30 s over 10 min and GR activity was calculated from the reduced GSSG concentration using the extinction coefficient 6.2 mM -1 cm -1 (Murshed et al 2008). NOEC no observed effect concentration; EC 80 concentration causing 80% of growth inhibition.…”

Section: Enzyme Assaysmentioning

confidence: 99%

Biological responses of maize (Zea mays) plants exposed to chlorobenzenes. Case study of monochloro-, 1,4-dichloro- and 1,2,4-trichloro-benzenes

et al. 2011

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A 7-day-exposure time experiment was designed to investigate the phytotoxicity of chlorobenzenes (CBs) on Zea mays seedlings, focusing on the growth and generation of oxidative stress. Significant growth inhibition (based on biomass gain) was observed for exposure to monochlorobenzene (MCB), dichlorobenzene (DCB) and trichlorobenzene (TCB) concentrations higher than 10 mg l(-1). It would seem that CBs inhibit cell division, since the mitotic index decreased for roots exposed to DCB at 80 mg l(-1) dose (8%) and to all the TCB concentrations tested (20% inhibition). CBs exposure resulting in an increase in the oxidative stress response in maize seedlings [reactive oxygen species like H(2)O(2), antioxidant enzymes (POD, GR), lipid peroxidation] correlated to the compound's degree of chlorination, where damage increasing with the number of chlorine atoms (MCB < DCB < TCB). This biological response was also dependent on the dose-exposure. Z. mays exposed to CBs at concentrations <10 mg l(-1) did not induce sufficient oxidative damage to cause root cell death. Therefore, CBs at current environmental concentrations are unlikely to produce evident phytotoxic effects on Z. mays seedlings.

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Section: Enzyme Assaysmentioning

confidence: 99%

Biological responses of maize (Zea mays) plants exposed to chlorobenzenes. Case study of monochloro-, 1,4-dichloro- and 1,2,4-trichloro-benzenes

et al. 2011

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“…Protein extraction was performed according to the Murshed et al (2008b) method. Frozen fruit powder (0.20 to 0.4 g) was homogenized in 1 ml of 50 mM MES/KOH buffer (pH 6.0), containing 40 mM KCl, 2 mM CaCl 2 , and 1 mM AsA.…”

Section: Extraction Of Enzymesmentioning

confidence: 99%

“…APX, DHAR, MDHAR and GR activities were measured as described in the method of Murshed et al (2008b). SOD activity was assayed in a 1.0 ml reaction mixture containing 50 mM potassium phosphate (pH 7.8) buffer, 13 mM methionine, 75 μM nitro blue tetrazolium (NBT), 0.1 mM EDTA, 10 μL of sample supernatant and 2 μM riboflavin.…”

Section: Enzyme Assaysmentioning

confidence: 99%

Effect of water stress on antioxidant systems and oxidative parameters in fruits of tomato (Solanum lycopersicon L, cv. Micro-tom)

Murshed

Lopez-Lauri²,

Sallanon

2013

Physiol Mol Biol Plants

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The effects of different levels of water stress on oxidative parameters (H 2 O 2 and MDA), the total pool sizes of ascorbate, the activities of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), as well as the activities and relative transcript levels of the enzymes of ascorbate-glutathione cycle ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) were studied in the fruit of tomato (Solanum lycopersicum L. cv. Micro-Tom). Plants were subjected to three levels of water stress (S50, S25 and S0) and fruits at different development stages were harvested after 3, 6 and 10 days of stress. Changes in H 2 O 2 and MDA contents indicated that water stress induced oxidative stress in fruits. The concentrations of ascorbate (AsA) and dehydroascorbate (DHA) generally modified with water stress treatments. Moreover, changes in SOD and CAT activities and DHAR, MDHAR, APX and GR activities and relative transcript levels were dependent on the fruit development stage and the intensity and the duration of water stress. These results suggest that the response of antioxidant systems of tomato fruits to oxidative stress induced by water stress treatments was different depending on the fruit development stage.

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“…Activities of APX (EC 1.11.1.11), DHAR (EC 1.8.5.1), MDHAR (EC1.6.5.4), GR (EC 1.8.1.7), and POX (EC 1.11.1) were measured as described by Murshed et al (2008) with slight modifications. Briefly, frozen leaf tissue (approximately 150 mg of pooled leaves 5, 6, and 7 from four different plants) was homogenized with the Retsch Mixer Mill MM2000 with two stainless-steel beads (2 mm diameter) in each sample.…”

Section: Analysis Of Enzyme Activitiesmentioning

confidence: 99%

A Novel Protective Function for Cytokinin in the Light Stress Response Is Mediated by the ARABIDOPSIS HISTIDINE KINASE2 and ARABIDOPSIS HISTIDINE KINASE3 Receptors

et al. 2014

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Cytokinins are plant hormones that regulate diverse processes in plant development and responses to biotic and abiotic stresses. In this study, we show that Arabidopsis (Arabidopsis thaliana) plants with a reduced cytokinin status (i.e. cytokinin receptor mutants and transgenic cytokinin-deficient plants) are more susceptible to light stress compared with wild-type plants. This was reflected by a stronger photoinhibition after 24 h of high light (approximately 1,000 mmol m 22 s 21), as shown by the decline in maximum quantum efficiency of photosystem II photochemistry. Photosystem II, especially the D1 protein, is highly sensitive to the detrimental impact of light. Therefore, photoinhibition is always observed when the rate of photodamage exceeds the rate of D1 repair. We demonstrate that in plants with a reduced cytokinin status, the D1 protein level was strongly decreased upon light stress. Inhibition of the D1 repair cycle by lincomycin treatment indicated that these plants experience stronger photodamage. The efficiency of photoprotective mechanisms, such as nonenzymatic and enzymatic scavenging systems, was decreased in plants with a reduced cytokinin status, which could be a cause for the increased photodamage and subsequent D1 degradation. Additionally, slow and incomplete recovery in these plants after light stress indicated insufficient D1 repair. Mutant analysis revealed that the protective function of cytokinin during light stress depends on the ARABIDOPSIS HISTIDINE KINASE2 (AHK2) and AHK3 receptors and the type B ARABIDOPSIS RESPONSE REGULATOR1 (ARR1) and ARR12. We conclude that proper cytokinin signaling and regulation of specific target genes are necessary to protect leaves efficiently from light stress.Light absorption, the subsequent conversion into biochemical energy, and the production of oxygen of plants play an essential role for life on earth. Although light is a prerequisite for this process, high light (HL) easily exceeds the plant's capacity to assimilate CO 2 , causing an overreduction of the electron transport chain that results in the inactivation of PSII (photoinhibition; Barber and Andersson, 1992; Aro et al., 1993;Yamamoto et al., 2008). One of the major targets of photoinhibition is the PSII core protein D1 (for review, see Adir et al., 2003;Edelman and Mattoo, 2008).Damaged D1 proteins are continuously replaced by de novo-synthesized D1 in a process called the D1 repair cycle (Aro et al., 1993(Aro et al., , 2005Baena-González and Aro, 2002). This cycle consists of (1) migration of damaged D1 protein from the grana to the stroma lamellae, (2) proteolytic degradation of damaged D1 protein by FILA-MENTATION TEMPERATURE SENSITIVE H (FTSH) protease and DEGRADATION OF PERIPLASMIC PRO-TEINS PROTEASE (DEGP), (3) de novo synthesis of precursor D1 protein (preD1) and cotranslational insertion into the thylakoid membrane, (4) C-terminal processing of preD1 catalyzed by the C-TERMINAL PEPTIDASE (CTP), and (5) migration to the grana thylakoids and formation of a fully functional PSII comple...

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Microplate quantification of enzymes of the plant ascorbate–glutathione cycle

Cited by 244 publications

References 9 publications

Biological responses of maize (Zea mays) plants exposed to chlorobenzenes. Case study of monochloro-, 1,4-dichloro- and 1,2,4-trichloro-benzenes

Biological responses of maize (Zea mays) plants exposed to chlorobenzenes. Case study of monochloro-, 1,4-dichloro- and 1,2,4-trichloro-benzenes

Effect of water stress on antioxidant systems and oxidative parameters in fruits of tomato (Solanum lycopersicon L, cv. Micro-tom)

A Novel Protective Function for Cytokinin in the Light Stress Response Is Mediated by the ARABIDOPSIS HISTIDINE KINASE2 and ARABIDOPSIS HISTIDINE KINASE3 Receptors

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