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Proceeding of the 4 rd International Symposium“NEW RESEARCH IN BIOTECHNOLOGY” USAMV Bucharest, Romania, 2011PEG on the first day. There was increase of its intensity corresponding to the increase ofPEG concentration. It was most illustrative on the sixth day (lanes 8-10) and on the twentythird day (lanes 11-13). Isoperoxidase profile of soybean in vitro cultures treated withdifferent concentrations of PEG for thirty days is presented in Fig 3. Excluding isoform 10,a similar pattern was observed. Isoform 35 was not found in the control. Consideringisoforms 21, 24, 28 and 35, the most intense response to the stress factor was recorded incalli cultures (lanes 1-3).The exposure of soybean shoot cultures to osmotic (drought) stress, simulated by PEG,induced new isoperoxidases as a response to the stress factor. Induction of peroxidase havebeen observed also in Aegilops (Landjeva et al., 2004), barley (Tamás et al., 2007) andHevea brasiliensis (Dutsadee and Nunta, 2008).Differential response of soybean peroxidase to the stress factor was recorded dependingon PEG concentration and time treatment. Generally, there was gradual increase ofisoperoxidase intensity (activity) which corresponded to the increase of PEG concentration.Similar pattern of gradual increase of isoperoxidases was found in Aegilops (Landjeva etal., 2004) too. Analysis of 13 POD genes in sweetpotato revealed differential expression ofPODs in response to the stress factor (Yun-Hee Kim et al., 2010).Fig 3. Isoperoxidase profile of soybean in vitro cultures treated with different concentrations ofPEG for 30 days.Calli: 1 – 4 % PEG; 2 – 6 % PEG; 3 – 8 % PEG; 4 – 0 % PEG (Control);Shoots: 5 – 0 % PEG (Control); 6 – 4 % PEG; 7 – 6 % PEG; 8 – 8 % PEG4. CONCLUSIONIt could be concluded that induction, increase of activity and differential expression inresponse to the stress factor are closely linked to an improved antioxidant defense capabilitymediated by peroxidase in soybean cv Rosa. This variety was chosen in these experiments forits higher yield under non-irrigated field conditions (Todorova and Goranova, 2010) and isconsidered with higher drought tolerance.12
Proceeding of the 4 rd International Symposium“NEW RESEARCH IN BIOTECHNOLOGY” USAMV Bucharest, Romania, 2011Presented results enrich our previous research on comparison between the response ofvarious soybean lines to water deficiency in field and laboratory conditions using droughtsimulation by PEG. A tendency of positive correlation between seed yield from one side andseed germination and seedling growth stages from another was observed (Kosturkova et al.,2008). Cotyledonary nodal explants subjected to 6% PEG 6000 treatments, resulted inreduction in shoot induction, compared to the control. In vitro callus cultures of both Indianand soybean cultivars grown in Bulgarian showed a reduction in callus growth with PEGtreatment, compared with the control. Presence of PEG in the medium elevated dry mattercontent in all the treatments compared to the control (Saktivelu et al., 2008b).ACKNOWLEDGEMENTS. Auhtors thank to Dr. R. Todorova, associate professor of theSoybean Experimental Station – Pavlikeni for providing soybean seeds.REFERENCESBajij, M., Lutts S., Kinet J. M. (2000): Physiological changes after exposure to and recovery frompolyethylene glycol-induced water deficit in callus culture issued from durum wheat (Triticum durum)cultivars differing in drought resistance, J. Plant Physiol. Vol. 156: 75–83.Boyer, J.S. (1982): Plant productivity and environment. Science. Vol. 218: 443–448.Davis, B. (1964): Disc electrophoresis. I. Method and application to human serum proteins. Ann NY Acad Sc. Vol. 121: 404-427.Diab, A.A., Cantety, R.V., Osturk, N.Z., Benscher, D., Nachit M.M. and Sorrells, M.E. (2008):Drought – Inducible Genes and Differentially Expressed Sequence Tags Associated with Components ofDrought Tolerance in Durum Wheat. Scientific Research and Essay. Vol. 3, (1): 9-26.Dutsadee, Ch., and Nunta, Ch. (2008): Induction of peroxidase, scopoletin, phenolic compoundsand resistance in Hevea brasiliensis by elicitin and a novel protein elicitor purified from Phytophthorapalmivora. Physiological and Molecular Plant Pathology. Vol. 72: 179-187.Fang, V., Kao Ch. (2000): Enhanced peroxidase activity in rice leaves in response to excess Fe, Cuand Zn. Plant Science. Vol. 158: 71-76.Foyer, C., Descourvieres P., Kunert, K. (1994): Protection against oxygen radicals: an importantdefense mechanism studied in transgenic plants. Plant Cell Envirn. Vol. 17: 507-523.Goranova, K. and Todorova, R. (2005): Past, present and future of the breeding of soybean. In:Breeding and Technological Aspects in Production and Processing of Soybean and Other LegumeCrops”, ed. G. Georgiev, Pavlikeni, 30-41.Gorjizad, A., Livani B., Abadi, D. (2010): Study of water stress (flooding and drought) on someantioxidant enzymes and essence percent in Anthemis altissima L. J. Biotechnology. Vol. 150: 185-193.Kosturkova, G. (2005): In vitro development of various soybean (Glycine max) explants frommature seeds. “Breeding and Technological Aspects in Production and Processing of Soybean and OtherLegume Crops”. Scientific Reports of the Jubilee Scientific conference (with international participation),8-9 Sept. 2005 Pavlikeni, 94-99.Kosturkova, G., Nedev, T., Dimitrova, M. (2006): Application of callus cultures of soybean (Glycinemax) to study abiotic stress factors. Field Crops Studies (Bg). Vol. 3, (2): 245-249.Kosturkova, G., Todorova, R. , Sakthivelu G., Akitha Devi M. K., Giridhar, P., Rajasekaran, T.,Ravishankar, G. A. (2008): Response of Bulgarian and Indian soybean genotypes to drought and waterdeficiency in field and laboratory conditions. Journal of general and Applied Plant Physiology. Vol. 34(3-4): 239-25013
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