The pyrogallol assay for superoxide dismutase: Absence of a glutathione artifact

Röth, Elizabeth; Gilbert, Harriet S.

doi:10.1016/0003-2697(84)90344-0

Cited by 104 publications

(41 citation statements)

References 8 publications

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“…SOD assay. SOD activity was monitored according to a published method (Roth & Gilbert, 1984). One millilitre of reaction mixture contained 50 mM sodium phosphate buffer (pH 7.8), 100 mM EDTA, 20 ml enzyme extract and 10 mM pyrogallol.…”

Section: Methodsmentioning

confidence: 99%

Antioxidant enzyme activities in maize plants colonized with Piriformospora indica

et al. 2009

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The bioprotection performance of Piriformospora indica against the root parasite Fusarium verticillioides was studied. We found that maize plants first grown with F. verticillioides and at day 10 inoculated with P. indica showed improvements in biomass, and root length and number as compared with plants grown with F. verticillioides alone. To validate our finding that inoculation with P. indica suppresses colonization by F. verticillioides, we performed PCR analyses using P. indica-and F. verticillioides-specific primers. Our results showed that inoculation with P. indica suppresses further colonization by F. verticillioides. We hypothesized that as the colonization by P. indica increases, the presence of/colonization by F. verticillioides decreases. In roots, catalase (CAT), glutathione reductase (GR), glutathione S-transferase (GST) and superoxide dismutase (SOD) activities were found to be higher in F. verticillioides-colonized plants than in noncolonized plants. Increased activity of antioxidant enzymes minimizes the chances of oxidative burst (excessive production of reactive oxygen species), and therefore F. verticillioides might be protected from the oxidative defence system during colonization. We also observed decreased antioxidant enzyme activities in plants first inoculated with F. verticillioides and at day 10 inoculated with P. indica as compared with plants inoculated with F. verticillioides alone. These decreased antioxidant enzyme activities due to the presence of P. indica help the plant to overcome the disease load of F. verticillioides. We propose that P. indica can be used as a bioprotection agent against the root parasite F. verticillioides. INTRODUCTIONMycorrhizal associations are among the most widespread, intimate and important symbioses in terrestrial ecosystems. Among these, arbuscular mycorrhiza (AM) represent the most ancient and widespread mycorrhizal symbiosis, and are found in a wide range of land plant species (Harrison, 1997). The successful establishment of this mutualistic association constitutes a strategy to fulfil the nutritional demands of both partners . This requires a balance between the defence responses of the host plant and the nutrient demands of the endophyte, resulting in an altered defence-related gene expression, which has been extensively studied during host colonization by obligate biotrophic arbuscular mycorrhizal fungi (AMF). Induction of defence-related genes is most prominent during the early stages of colonization (García-Garrido & Ocampo, 2002), but can also be detected during arbuscule development (Grunwald et al., 2004).The active resistance of plants to colonization by fungi is often expressed by the hypersensitive reaction (HR) of challenged plant cells, and is characterized by reactive oxygen species (ROS), as well as by induced rapid and localized death of plant tissue at the site of infection (Ingram, 1978;Tenhaken et al., 1995). At the biochemical level, the rapid generation of ROS, such as superoxide (O { 2 ) and hydrogen peroxide (H 2 O 2 ), kno...

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Section: Methodsmentioning

confidence: 99%

Antioxidant enzyme activities in maize plants colonized with Piriformospora indica

et al. 2009

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“…Dialysis eliminated redox components of low mol wt as ascorbic acid and glutathione, which are involved in nonenzymic detoxification of toxic oxygen species (12,21). These components are abundant in cells and interact with superoxide radicals (16,22). Therefore, they can interfere with assays for superoxide dismutase which rely on the participation of 2 as the chain propagating species.…”

Section: Discussionmentioning

confidence: 99%

Superoxide Dismutase Activity in Needles of Norwegian Spruce Trees (Picea abies L.)

Polle¹,

Krings²,

Rennenberg³

1989

Plant Physiol.

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The activity of superoxide dismutase was investigated in needles of spruce trees. To obtain maximum activity, needles were homogenized in the presence of Triton X-100 and polyvinylpyrrolidone. Superoxide dismutase activity was measured in dialyzed extracts with a modified epinephrine assay (HP Misra, IFridovich (1972] J Biol Chem 247: 3170-3175) at pH 10.2. The extracts contained 70 to 120 units of superoxide dismutase per milligram protein. One unit of superoxide dismutase was completely inhibited in the presence of 20 micromolar NaCN. On nafive polyacrylamide gels three electromorphs were visualized after staining for activity. All three species were sensitive to CN-and H202 and were therefore assumed to be Cu/Zn-superoxide dismutases. Superoxide dismutase activity was dependent on the age of the needles and declined by approximately 25% within 3 to 4 years.

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“…SOD activity was monitored according to the method from Roth & Gilbert (1984) and modified by Kumar et al (2009). One ml of reaction mixture contained 50 mM sodium phosphate buffer (pH 7.8), 100 mM EDTA, 20 ml enzyme extract, and 10 mM pyrogallol.…”

Section: Superoxide Dismutase (Sod) Assaymentioning

confidence: 99%

Tolerance to heavy metal stress in seedlings of three pine species from contrasting environmental conditions in Chile

Arencibia¹,

Rodríguez²,

Roco³

et al. 2016

iForest

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Tolerance to metal stress was studied in seedlings of three pine species (Pinus radiata, P. pinaster and P. canariensis) under controlled ex vitro conditions. Mature female cones were randomly collected at two sites in Chile (Llico and Huilquilemu) characterized by contrasting environmental conditions. Oneyear-old pine seedlings were immersed in trays with solutions of CuSO4 (300 mM) or AlCl3 (100 mM), and their survival, growth rate and decay symptoms were recorded for 60 days. Results showed large differences among provenances in seedling tolerance to CuSO4 and AlCl3 in terms of survival and growth. Multivariate analysis revealed a significant association (p <0.0001) between the first canonical function and the following variables: provenance, species, metal stress and growing rate, as well as between the second canonical function and provenance, species, metal stress and symptomatology, indicating a high degree of genotype-environment interaction. Moreover, the activity of POX, SOD and CAT enzymes was determined 60 days after the beginning of the experiment in the Llico provenance seedlings. P. pinaster showed the highest activity level for all the enzymes considered, while P. canariensis and P. radiata had intermediate and lowest values, respectively. Differential gene expression among pine seedlings under metal stress with CuSO4 for two genes (Cu-Zn-superoxide dismutase and RuBisCo) confirmed P. pinaster as the most tolerant species to CuSO4 treatment. Our results are consistent with the hypothesis that abiotic stress in the maternal environment can induce a "transgenerational plasticity" which could affect progeny performances. The influence of different genetic backgrounds on the tolerance to heavy metals in pine seedlings is also discussed.

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The pyrogallol assay for superoxide dismutase: Absence of a glutathione artifact

Cited by 104 publications

References 8 publications

Antioxidant enzyme activities in maize plants colonized with Piriformospora indica

Antioxidant enzyme activities in maize plants colonized with Piriformospora indica

Superoxide Dismutase Activity in Needles of Norwegian Spruce Trees (Picea abies L.)

Tolerance to heavy metal stress in seedlings of three pine species from contrasting environmental conditions in Chile

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