The effect of nitric oxide (NO) on antioxidant system and protective mechanism against oxidative stress under UV-B radiation was investigated in leaves of maize (Zea mays L.) seedlings during 3 days growth period. UV-B irradiation caused a decrease of leaf biomass including leaf length, width and weight during growth. Application of NO donor, sodium nitroprusside (SNP), significantly alleviated UV-B stress induced growth suppression. NO donor permitted the survival of more green leaf tissue preventing chlorophyll content reduction and of higher quantum yield for photosystem II than in non-treated controls under UV-B stress, suggesting that NO has protective effect on chloroplast membrane in maize leaves. Flavonoids and anthocyanin, UV-B absorbing compounds, were significantly accumulated in the maize leaves upon UV-B exposure. Moreover, the increase of these compounds was intensified in the NO treated seedlings. UV-B treatment resulted in lipid peroxidation and induced accumulation of hydrogen peroxide ($H_2O_2$) in maize leaves, while NO donor prevented UV-B induced increase in the contents of malondialdehyde (MDA) and $H_2O_2$. These results demonstrate that NO serves as antioxidant agent able to scavenge $H_2O_2$ to protect plant cells from oxidative damage. The activities of two antioxidant enzymes that scavenge reactive oxygen species, catalase (CAT) and ascorbate peroxidase (APX) in maize leaves in the presence of NO donor under UV-B stress were higher than those under UV-B stress alone. Application of 2-(4-carboxyphenyl)-4, 4, 5, 5-tetramethylimidazoline-1-oxyl-3- oxide (PTIO), a specific NO scavenger, to the maize leaves arrested NO donor mediated protective effect on leaf growth, photosynthetic pigment and free radical scavenging activity. However, PTIO had little effect on maize leaves under UV-B stress compared with that of UV-B stress alone. $N^{\omega}$-nitro-L-arginine (LNNA), an inhibitor of nitric oxide synthase (NOS), significantly increased $H_2O_2$ and MDA accumulation and decreased antioxidant enzyme activities in maize leaves under UV-B stress. This demonstrates that NOS inhibitor LNNA has opposite effects on oxidative resistance. From these results it is suggested that NO might act as a signal in activating active oxygen scavenging system that protects plants from oxidative stress induced by UV-B radiation and thus confer UV-B tolerance.
Kim, Dea-Wook;Heo, Hwa-Young;Suh, Sae-Jung;Lee, Yeong-Ho;Kim, Si-Ju
KOREAN JOURNAL OF CROP SCIENCE
/
v.51
no.spc1
/
pp.133-138
/
2006
To differentiate barley responses to drought and salt stress, barley seedlings at the second leaf stage were treated with 218 mM NaCl and 29.5% PEG6000 iso-osmotic to 218 mM NaCl for 6 days. Shoot fresh weight and leaf relative water content of barley seedlings were more reduced by drought compared to salt stress. Hydrogen peroxide content increased under both stress conditions, but its accumulation was more severe at 6 days after salt stress. The activity of ascorbate peroxidase, glutathione reductase (GR) and catalase (CAT) was enhanced until 4 days after salt stress. On the other hand, the activity of GR and CAT increased gradually until 6 days after drought. Among the amino acids measured in this study, the accumulation of glycine, arginine and GABA (${\gamma}-aminobutyric$ acid) was lower under salt stress than drought. However, considerably larger amount of proline was accumulated by salt stress. It is concluded that the antioxidant enzymes activity and amino acid content of barley seed-lings were differently regulated in response to the isoosmotic condition of salt and drought stress.
In order to observe the bioactivity of ovariectomized rats, ovariectomized group (Ovx), nonovariectomized group (Sham), ovariectomized Vitamin C-treat group (Ovx+Vit C), ovariectomized Vitamin E-treat group (Ovx+Vit E) and ovariectomized Vitamin C+Vitamin E-treat group(Ovx+Vit C+E) were made. Lipidperoxides of liver and kidney, serum total cholesterol and HDL-cholesterol were investigate as follows. Lipidperoxides of liver and kidney in Ovx group were 1.78 times and 1.61 times increased compared to Sham group respectively. But, they were significantly decreased in Ovx+Vit C group, Ovx+Vit E group, Ovx+Vit C+E group compared to Ovx group. Serum total cholesterol in Ovx group was increased 2.57 times compared to Sham group. Injections of each substance such as ascorbate, tocopherol, mixture (C+E) make data of Cholesterol become low. When especially Vit C is injected, the data of cholesterol lowed by about 94%. Serum HDL-cholesterol in Ovx group decreased 36.7% compared to Sham group. And as the result of the measurement of SOD, Catalase, and GPx which are antioxidant enzyme, SOD and Catalase activities in Ovx group much higher than in Sham group. Based on the results, it is supposed that more produced free radicals increased antioxidant enzyme. And it is also thought that vitamin can inhibit aging by reducing antioxidant enzyme.
A Corynebacterium diphtheriae iron-repressible gene dirA, that was homologous to TSA of Saccharomyces cerevisiae and AhpC subunit of Salmonella typhimurium alkyl hydroperoxide reductase, was amplified with PCR and expressed in E. coli. The DirA purified from the transformed E. coli crude extracts prevented the inactivation of enzyme caused by metal-catalyzed oxidation (MCO) system containing thiols but not by ascorbate/Fe$^{3+}$/$O_2$ MCO system. The DirA concentration, which inhibited the inactivation of glutamine synthetase by 50% (IC$_{50}$) against MCO system, was 0.12 mg/ml. The multimeric forms of DirA were converted to the monomeric form in SDS-PAGE under the thioredoxin system comprised of NADPH, Saccharomyces cerevisiae thioredoxin reductase, and thioredoxin. Also, DirA showed thioredoxin dependent peroxidase activity. All of these results were consistent with the characteristics of a thiol specific antioxidant (TSA) protein having two conserved cysteine residues.
Proceedings of the Korean Society of Plant Biotechnology Conference
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2003.04a
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pp.65-71
/
2003
Bax, a mammalian pro-apoptotic member of the Bcl-2 family, induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorho-damine 123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MBP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to play a novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.
Ozone sensitivity of physiological indicators and the difference of ozone tolerance on 4 families of Quercus aliena seedlings were investigated on the basis of the standardized physiological indicators. Photosynthetic parameters, photosynthetic pigment and malondialdehyde (MDA) content, and antioxidative enzyme activities were measured or analyzed from the leaves of Q. aliena seedlings at the end of ozone fumigation, and ozone tolerance indices among 4 families were calculated with the standardized physiological parameters. After ozone treatment, the reduction of carboxylation efficiency was observed in the leaves of four families, and their reduction were ranged from -24.1% to -56.9% of control seedlings. Photosynthetic pigment content differed significantly among 4 families and treatments. The reduction of total chlorophyll content showed the highest in family SU4 (-40.6%) and the lowest family US2 (-18.8%). Ascorbate-peroxidase (APX) activity showed significant difference among families and treatments, and increased as compared with control in three families, except for family US2. On the basis of the physiological indices, ozone tolerance of four families was ranked in the order of US1 > SU4 > US2 > SU1. In conclusion, photosynthetic parameters, pigment content and APX activity were recommended as appropriate indicators to assess the tolerance against ozone stress of Q. aliena.
Antioxidative enzymes (superoxide dismutase; SOD, ascorbate peroxidase; APX, glutathione reductase; GR) play major roles in scavenging mechanism of reactive oxygen species which were involved in various stress conditions including salt. In order to investigate the relation between their growth responses (dry weight) and the changes of antioxidative enzymes activity, salt-tolerant spinach beet having 15cm of shoot length were treated with various salt levels (0, 50, 200, 1000 mM NaCl) for 24 hours. Spinach beet exhibited an increase in the activity of antioxidative enzymes by salt, the maximal activity at 200 mM NaCl and the lowest activity at 50 mM NaCl in 2 hrs. after treatments. As a result of PAGE, it has been confirmed that spinach beet contained 3 isoforms (Fe-SOD, CuZn-SOD and Mn-SOD) of SOD and main isoform was CuZn- SOD form. In case of APX, isoforms of the low molecular weight(No. 7, 8) were showed strong expression especially at 200 and 400 mM NaCl treatment. Meanwhile, GR did not show specific pattern of isoforms among the salt treatments. Especially, in case of 50 mM treatment, plant showed the lowest activity of SOD with the best growth, a low enzyme activity was induced by inactivation of the Mn-SOD. Therefore, we suggested that the decrease of SOD activity at a low salt level (50 mM NaCl) or the increase of enzyme activity at a high salt level (200 mM NaCl) may be related to expression of the Mn-SOD isoform. These antioxidative enzymes showed the increase of activity in a short time by salt addition. So, it is considered that spinach beet copes effectively with a stressful condition such as salt by operating effective antioxidative defense mechanism rapidly under high salt level.
Kim, Yoonha;Waqas, Muhammad;Khan, Abdul Latif;Mun, Bong-Gyu;Yun, Byung-Wook;Lee, In-Jung
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.203-203
/
2017
The Earth's climate is rapidly changing because of increasing carbon dioxide content in atmosphere so, climate prediction models anticipate that earth surface temperature will rise by 3 to $5^{\circ}C$ in next 50 to 100 years. Therefore, frequency of un-expected weather events such as drought, salinity, low or high temperature and flooding etc. will be increasing worldwide. Furthermore, increased atmosphere temperature can influence pests and pathogens spread as well. Therefore, to protect enormous grain loss from unexpected weather conditions, studies related with combine stress conditions like abiotic plus biotic stress condition are really required. Thus, our research focused on physiological responses under combined abiotic and biotic stress condition in rice plant. To induce uniform stress condition, we used NaCl (100 mM) and salicylic acid (0.5 and 1.0 mM SA) as each stress a stimulator. Each artificial abiotic and biotic stress inducer was applied to hydroponically grown rice seedlings alone or together for four day. The data were collected in a time-dependent manner [1, 2, 3 and 4 day(s) after treatment (DAT)] and were matched with our anticipation that shoot length and shoot fresh weight was decreased in solo and combined abiotic and biotic stress condition. The lipid peroxidation content was significantly increased ($1.5{\pm}0.2$ to $2.7{\pm}0.1mg$ mg of $MDA\;g^{-1}FW$) in the first two days in both stress exposed plants, and showed the opposite trend ($0.5{\pm}0.01$ to $0.1{\pm}0.001mg$ of $MDA\;g^{-1}FW$) in last two days under multi stress condition. Superoxide dismutase (SOD) activity did not showed difference in only biotic stress condition (alone 0.5 and 1.0 mM SA) as compared to control however, it was significantly increased in multi stress condition or solo abiotic stress condition whereas, catalase (CAT), and ascorbate peroxidase (APX) activities were significantly decreased in solo biotic and combined abiotic and biotic condition. In particular, both enzymes activities were more decreased in multi stress condition as compared to solo biotic stress condition. The results for relative mRNA expression level of CAT and APX enzymes were in agreement with results of spectrophotometric values. Correlation value between each stress condition and phenotypic data showed that biotic stress condition showed high correlation with activity of CAT and APX whilst, abiotic stress condition revealed significant correlation with SOD activity.
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.127-127
/
2017
Aluminum is the most abundant metallic element in the Earth's crust and considered as the most limiting factor for plant productivity in acidic soils. The inhibition of root growth is recognized as the primary effect of Al toxicity. Seeds of wheat cv. Keumkang (Korean cultivar) were germinated on petridish for 5 days and then transferred hydroponic apparatus which was treated with $0{\mu}M$$AlCl_3$ (control), $100{\mu}M$$AlCl_3$ and $150{\mu}M$$AlCl_3$ for 5 days. The length of roots, shoots and fresh weight of wheat seedlings were decreased under aluminum stress. The concentrations of $K^+$, $Mg^{2+}$ and $Ac^{2+}$ were decreased whereas $Al^{3+}$ and $P_2O_5{^-}$ concentration was increased under aluminum stress. Using confocal microscopy, the fluorescence intensity of aluminum was increased with morin staining. In this study, a proteome analysis was performed to identify proteins, which is responsible to aluminum stress in wheat roots. In 10-day-old seedlings, proteins were extracted from roots and separated by 2-DE, stained by CBB. Using image analysis, a total of 47 differentially expressed protein spots were selected, whereas 19 protein spots were significantly up-regulated such as s-adenosylmethionine, oxalate oxidase, malate dehydrogenase, cysteine synthase, ascorbate peroxidase and 28 protein spots were significantly down-regulated such as heat shock protein 70, o-methytransferase 4, enolase, amylogenin by aluminum stress following protein spots analyzed by LTQ-FTICR mass spectrometry. The results provide the global picture of Al toxicity-induced alterations of protein profiles in wheat roots, and identify the Al toxicity-responsive proteins related to various biological processes that may provide some novel clues about plant Al tolerance.
Kim, Du-Hyun;Han, Sim-Hee;Ku, Ja-Jung;Lee, Kab-Yeon;Kim, Pan-Gi
Korean Journal of Agricultural and Forest Meteorology
/
v.10
no.2
/
pp.47-57
/
2008
Physiological and biochemical changes of five species of genus Quercus exposed to ozone fumigation were investigated to assess their tolerance against ozone toxicity. At the end of 150 ppb ozone fumigation, chlorophyll contents, photosynthetic characteristics, malondialdehyde(MDA) and antioxidative enzyme activities were measured in the leaves of five Quercus species(Quercus acutissima, Q. aliena, Q. palustris, Q. serrata, and Q. variabilis). Chlorophyll and carotenoid contents, net photosynthesis and carboxylation efficiency decreased after ozone treatment, indicating that $O_3$-exposed plants underwent physiological inhibition. The reduction rate of total chlorophyll contents and carboxylation efficiency were respectively 15% and 34% for Q. aliena and 38% and 62% for Q. variabilis. The amount of MDA increased with the highest increase rate of 140% in Q. acutissima which also showed the highest increase rate(60%) of superoxide dismutase(SOD). Ascorbate peroxidase(APX) activity increased in Q. variabilis, Q. serrata and Q. acutissima by ozone treatment. Based on our results, ozone tolerance of the five Quercus species was ranked as Q. aliena>Q. palustris>Q. serrata>Q. variabilis>Q. acutissima. We concluded that chlorophyll contents, photosynthesis, MDA content and antioxidative enzymes were the important physiological markers for tolerance against ozone stress, which were closely related with one another.
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