Middle-East Journal of Scientific Research 7 (1): 07-11, 2011ISSN 1990 - PDF document

Middle-East Journal of Scientific Research 7 (1): 07-11, 2011ISSN 1990-9233© IDOSI Publications, 2011Corresponding Author:Mohammad Zaefizadeh, Islamic Azad University, Ardabil Branch, Ardabil, Iran.E-mail: mzaefi@gmail.com. Superoxide Dismutase (SOD) Activity in Nacl Stress in Salt-Sensitive and Salt-Tolerance Genotypes of Colza (Brassica napusSeyed Mohammad Shahab Jalali-e-Emam, Bahram Alizadeh, Mohammad Zaefizadeh, Superoxide Dismutase NaCl stress Antioxidant defense systemINTRODUCTION Uptake in control-Sample uptakePercent inhibition of resuscitation = Uptake in control-Distilled water uptakeMiddle-East J. Sci. Res., 7 (1): 7-11, 2011withincreasing concentration of ions Na and Clinrather than the potassium phosphate buffer was added.Cassia angustifolia seedlings were reported. The resultsThefirst solution to control any light away from theof research conducted on tomato [16], wheat [13, 17], beetradiation and the second was distilled water as controland cotton [12, 14] a significant increase in the Superoxidewithout light absorption were used and the absorptionDismutase enzyme activity in tolerant cultivars thanoflight at 560 nm by spectrophotometer shomadzosusceptiblecultivars under salt stress shows said. Also,samples were tested. Uptake in control solution as 100%complementarystudies significant differences betweenabsorption was zero value in reducing absorption in thethe secretion of this enzyme in salinity in the root to thesample was recorded. 50% of NBT reduction as a unit ofstemand leaves showed.Superoxide Dismutase activity intended SuperoxideThis Experiment aimed to study the activity ofDismutase activity was obtained by the following formula:antioxidant system Superoxide Dismutase enzyme andsecretion in sensitive and resistant canola varieties tosalinity was performed.MATERIALS AND METHODSRESULTS AND DISCUSSIONAmongthe 20 genotypes of canola, by seedAmount of measured Superoxide Dismutase usinggermination test on two different levels of salinity, twospectra photo metric in 560 nm wavelength meter showedgenotypesof colza, Quantum as a salt tolerant and Fornixthat between salinity levels of the enzyme Superoxideas a sensitive, were chosen. The seeds were planted inDismutase level was a significant difference in the 0.06Petri dishes in 27°C temperatures to 14 days Period at zeropercent level is observed. Thus, different levels of salinitysalinityconditions, 50, 100 and 150 mm Nacl. Thein the amount of Superoxide Dismutase in roots andgerminated seeds with roots and shoot in Petri dishshoots vary between genotype and environment (salinitychoice and shoot them separately and according to thelevels) a significant difference in the 0.01 percent levelfollowing method of enzyme extracted. Selected organswas observed. Thus, in sensitive and resistant cultivarswere powdered in liquid nitrogen and immediatelyto salinity differences significant amount of Superoxidewere kept in eppendorf tube at -20°C. Then 0/5 gramsDismutase has been. Comparison of enzyme in root andof frozen powdered root and shoot, in 10 ml of potassiumshoot showed different levels of salinity (Table 1) withphosphate buffer 50 mM with PH = 7 containing 1 mMincreasing salinity in the amount of Superoxide DismutaseEDTA, 1% poly and poly vinyl poly pyrrolidone and wasin roots and shoots is increased. As the highest level ofHomogenized. The Homogenized samples by icemanSuperoxide Dismutase in the root was in 150mM SalinityCentrifuge for 20 min in 4 Temperature with ×15000gr waslevels and root was in 100mmol. Purpose, can be statedcentrifuged.And Obtained super netEnt as enzymethatthe amount of salt in order to create stress amount ofsourcewas separated by sampler for next steps. The Rateenzyme secreted Superoxide Dismutase in stem and rootsof Superoxide Dismutase activity, were determined bywere different. The amount of Superoxide Dismutase inphotochemical nitro blue tetrazolium test method Beyerroots secretion was higher than shoot. Comparingand fridovich [18,19]. In this method, Protect NBTresistant and susceptible cultivars in terms of the amountreduction by extracted enzyme was determined by spectraofSuperoxide Dismutase indicated that Superoxidephoto metric method. thus, Identify solution ContainingDismutase in roots and shoots of resistant cultivars than27 ml 50 mM potassium phosphate solution with PH = 7/8,1/5 ml L-methionine, 1 ml nitro blue tetrazolium and 1000mlTriton - X 100 was prepared. One ml of this solution waspouredinto small tubes and then the 20 microliters ofExtracted enzyme was with 10 micro liter of riboflavin (4 /4 mg in 10 ml) was added. The solution 7 minutes withinthe chamber that completely covered by aluminum foil andthe two 20-watt fluorescent lamp was placed. Also, thetwo solutions as the control were considered. One ofthesesolutions, contain all the substances in solutionwereidentified and only the same amount of enzymeTable 1:Averageamount of enzyme activity in root and shoot inhibitionof regeneration at different levels of salinity (in terms of percentageinhibition of CPR)Salinity (Mmol)PlumuleRootControl22.9113.6733.9132.25 SE0.00260.0039 1234Salinity levels (mmol) Preventing regeneration (%) Quantom Fornax 1234Salinity levels (mmol) Preventing regeneration (%) Quantom FornaxMiddle-East J. Sci. Res., 7 (1): 7-11, 2011Fig. 1: Percent inhibition of nitro blue tetazolium reduction, in the salinity level in the roots of two cultivarsFig. 2: Percent inhibition of nitro Blue tetazolium reduction, in the salinity level the shoots of two cultivarsthe susceptible cultivar was. Considering the existence ofthan resistant cultivars significantly in root and shootsignificant interaction genotypes in salt levels, analysis ofwere higher. variance and mean for salt levels in both resistant andIthas been frequently reported that salt stresssusceptible varieties were carried. The results of thisinduces oxidative damage to plant tissues [13]. Thecomparison chart are the number one point. In The root,oxidative stress is considered to be due to increasedenzyme Superoxide Dismutase in the amount of resistantproduction of O, OH, HO and O. Hence, constitutivecultivars under salt stress significantly increases showed.and/or induced SOD is needed to prevent plant tissueSo that the amount of uptake inhibition in thefrom the oxidative damage. The tolerant genotypeconcentration0 mmol, 35% and 100 mmol 43% and(Quantum) had higher constitutive levels of SOD150mmol showed 50%. In both figure in normal conditionsactivities than the sensitive genotype (Table 2). Thistheamount of control there was little enzyme thatindicates that Quantum has potentially a better protectionresistantvarieties were slightly more susceptible.system against the oxidative damage caused by salinityInSensitive genotype, amount of enzyme in the rootstress. Several previous studies have also reported thatin stress condition any of the levels was not significant,salt-tolerant cultivars of tomato and beet had highermeaning that the rate of enzyme secretion in all conditionsconstitutive levels of antioxidant enzymes [20, 21].showed no significant change is.ByNaCl treatment, SOD activity was enhanced inIntolerant and resistant cultivars Shoots, Amount ofroot of Quantum, However the activities of SOD in root ofenzyme under stress conditions was significantlyFornax has not been changed significantly by increase inincreased (Fig. 1). But the reaction of susceptible andsaltlevels (Figure 1). These results are the same asresistantcultivars showed salinity levels shoot bothreported by Nur Cicerali [7] and Rios-Gonzalezet al. [22].cultivars with increasing salt concentrations up to 100Also,analysis of the SOD activity in shoots subjected tommol increase the absorption of preventing the existenceNaCl stress up to 100 mM NaCl revealed an increase inof an indicator enzyme is observed, but the concentrationSOD activity of both genotypes, but increase in saltof 150 mmol salt amount of enzyme than the enzymetolerant was higher than salt sensitive genotype (Fig. 2).concentration of 100 mmol amounts was reduced in bothHigh SOD activity was observed in shoot of tolerantcultivars.Reduce the amount of enzyme in susceptiblegenotypes of onions [23].222 Middle-East J. Sci. Res., 7 (1): 7-11, 2011SODactivity was enhanced in roots and shoots of5.Apel, K. and H. Hirt, 2004. Reactive Oxygen species:Quantum in relation to increase in salt concentration. TheMetabolism,oxidativestress and signal transduction.activity of SOD was also induced in shoots ofFornaxAnnu. Rev. Plant Biol., 55: 373-399.although its constitutive activity was lower than those of6.Hernandez, J.A., A. Compilo, A. Jimenez, J.J. AlarconQuantum(Table 1). So it is suggested that this enzymeandF. Sevilla, 2004. Responses of antioxidantfulfills the important role for the tolerance to NaClsystemsrelations to NaCl in plants. New physiol.,especially in Quantum as the same as reported in several152: 141-145.studies [13, 24].7.Nur-Ichik,A., 2004. Effect of NaCl stress onIn enzymatic scavenging systems, SOD is well knownantioxidantdefense system in lentil. M.Sc. Thesis,as O scavenger. The combined action of SOD convertsIstanbul university, Turkey. the potentially dangerous O and HO to water (HO) and8.Elstner, E.F., 1996. Mechanism of plant cell.222molecular oxygen (O) [1].In:E.J. Pell and K.L. Stefen (Eds), ActiveResultsof this study suggested that the highoxygen/oxidativestress and plant metabolism.constitutive and induced levels of SOD in the roots ofAmerican society of plant physiology, Rockville,Quantum. This may indicate that their combined action ispp: 13-25.aneffective scavenging mechanism to abate the toxic of9.Alscher,R.C., J.L. Donhae and C.L. Cramer, 1997.O in the root cells.Reactiveoxygen species and antioxidants:CONCLUSION100: 224-233.Results of this study clarified the differences of SuperJ.Cambraia, 2003. Photosynthesis and activity ofOxideDismutase activity betweenQuantum (as a saltsuperoxide dismutase, peroxidase and glutathionetolerant) and Fornax (as a salt sensitive). A high level ofreductasein cotton under salt stress. Environ. Exp.Super Oxide Dismutase, either constitutive or induced,Bot., 49: 69-76.has been reported to correlate with plant resistance to salt11.Mittler,R., 2002. Oxidative stress, antioxidants andstress.stress tolerance. Trends Plant Sci., 7: 405-410.Inaddition the SOD activity increasing as a result of12.Bowler,C., W.V. Camp, M.V. Montago and D. Inze,salt stress was stronger in the salt tolerant genotype1994.Super oxide dismutase in plants. Critical Rev.compared to the salt sensitive one. Results obtainedPlant Sci., 13: 199-218.support the hypothesis the higher efficiency of the13.Hernandez, J.A., A. Compilo, A. Jimenez, J.J. Alarconantioxidantenzymatic system ofQuantum (tolerant)andF. Sevilla, 1999. Responses of antioxidantgenotype could be considered as one of the factorssystemsand leaf water relations to NaCl in pearesponsiblefor its tolerance to salt stress. Therefore, it isplants. New physiol., 141: 241-251.suggested that superoxide dismutase activity could be14.Mckersie,B.D. and Y.Y. leshem, 1994. Stress andused as a working hypothesis for a biochemical marker forstresscoping in cultivated Plants. Kluwer academicsalt tolerance in rapeseed.publishes. 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