UV-C Stresinin 6-Benzilaminopürin Varlığında Çoğaltılmış Amsonia orientalis’in In vitro Sürgün Gelişimi ve Antioksidan Enzimleri Üzerine Etkileri

Yıl 2018, Cilt: 18 Sayı: 1, 44 - 52, 30.04.2018

Yonca Yüzügüllü Karakuş , Halil İ. Toygar Arda Acemi Yonca Avcı Duman Fazıl Özen

Öz

Bu çalışmada abiyotik faktörlerden biri olan UV-C stresinin 6-benzilaminopürin varlığında çoğaltılmış Amsonia orientalis’in in vitro sürgünleri üzerindeki etkileri, sürgünlerin morfometrik özellikleri, H2O2 (hidrojen peroksit) ve MDA (malondialdehit) seviyeleri ile antioksidan enzimleri analiz edilerek irdelenmiştir. Test edilen 3,47 (15 dk), 6,94 (30 dk) ve 13,87 (60 dk) kJ m-2’lik UV-C radyasyon dozlarının hepsi sürgün uzunluğu ile sürgün sayısında kontrole (UV-C uygulanmamış bitki) oranla düşüşe sebep olmuştur. Radyasyon kaynaklı oluşan bu stres 6-benzilaminopürin varlığında sürgün gelişimini yavaşlatsa da tamamen durdurmamıştır. UV-C radyasyon uygulamaları antioksidan enzimler üzerine farklı etkiler göstermiştir. SOD (süperoksit dismutaz) aktivitesi artan radyasyon dozuna bağlı olarak kontrole göre önemli ölçüde azalırken, POD (peroksidaz) aktivitesinde artış gözlenmiştir. CAT (katalaz) aktivitesinin ise radyasyondan önemli ölçüde etkilenmediği görülmüştür. Artan UV-C dozlarında H2O2 ve MDA seviyelerinin benzer olarak arttığı bulunmuştur. Elektroforetik analizler sonucunda SOD enzimi için üç farklı izozim (Cu/Zn-SOD, Fe-SOD ve Mn-SOD) tespit edilmiş olup, artan UV-C stresinin bitkinin Cu/Zn-SOD aktivitesini tamamen inhibe ettiği gözlenmiştir. Sonuçlar bitkinin UV stresine karşı gösterdiği savunma mekanizmasında peroksidazların incelenen diğer antioksidan enzimlere göre öncelikli rol aldığına işaret etmektedir.

Anahtar Kelimeler

Amsonia orientalis, katalaz, peroksidaz, süperoksit dismutaz, UV stresi

Kaynakça

• Acemi, A., Duman, Y., Karakuş, Y.Y., Kömpe, Y.Ö., Özen, F. 2017. Analysis of plant growth and biochemical parameters in Amsonia orientalis after in vitro salt stress. Horticulture, Environment, and Biotechnology, 58, 231-239.
• Acemi, A., Özen, F., Kıran, R. 2012. Development of an efficient callus production protocol for Amsonia orientalis: A critically endangered medicinal plant. Eurasian Journal of Biosciences, 6, 105-112.
• Acemi, A., Özen, F., Kıran, R. 2013. In vitro propagation of Amsonia orientalis Decne. from nodal segments of adult plants. Propagation of Ornamental Plants, 13, 25-32.
• Aebi, H. 1974. Methods of enzymatic analysis. Bergmeyer HU, Ed, Catalase. Academic Press, New York, 673-675.
• Akyalçın, H., Özen, F., Dülger, B. 2006. Anatomy, morphology, palynology and antimicrobial activity of Amsonia orientalis Decne (Apocynaceeae) growing in Turkey. International Journal of Botany, 2, 93-99.
• Allen, D.J., Nogues, S., Baker, N.R. 1998. Ozone depletion and increased UV-B radiation: is there a real threat to photosynthesis. Journal of Experimental Botany, 49, 1775-1788.
• Beauchamp, C.O., Fridovich, I. 1973. Isozymes of superoxide dismutase from wheat germ. Biochimica et Biophysica Acta, 317, 50-64.
• Bornman, C.H. 1983. Possibilities and constraints in the regeneration of trees from cotyledonary needles of Picea abies in vitro. Physiologia Plantarum, 57, 5-16.
• Bradford, M. 1976. A rapid and sensitive method for the quantification of microgram quantities of protein using the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254.
• Caldwell M.M., Björn L.O., Bornman J.F., Flint S.D., Kulandaivelu G., Teramura A.H., Tevini, M. 1998. Effects of increased solar ultraviolet radiation on terrestrial ecosystems. Journal of Photochemistry and Photobiology B: Biology, 46, 40-52.
• Chairat, B., Nutthachai, P., Varit, S. 2013. Effect of UV-C treatment on chlorophyll degradation, antioxidant enzyme activities and senescence in Chinese kale (Brassica oleracea var. alboglabra). International Food Research Journal, 20, 623-628.
• Chen, Y.P. 2009 Response of antioxidant defense system to laser radiation apical meristem of Isatis indigotica seedlings exposed to UV-B. Plant Signaling and Behavior, 4, 571-573.
• Darke, R. 2005. Amsonia in cultivation. The Plantsman, 4, 72-75.
• Darras, A.I., Demopoulos, V., Bali, I., Katsiloulis, O., Kratimenou, E. 2013. Brief exposures of ultraviolet-c (UV-C) irradiation improves flowering of ornamental plants. Acta Horticulturae, 1002, 95-101.
• Doupis, G., Chartzoulakis, K., Beis, A., Patakas, A. 2011. Allometric and biochemical responses of grapevines subjected to drought and enhanced ultraviolet-B radiation. Australian Journal of Grape and Wine Research, 17, 36-42.
• Erkan, M., Wang, S.Y., Wang, C.Y. 2008. Effect of UV treatment on antioxidant capacity, antioxidant enzyme activity and decay in strawberry fruit. Postharvest Biology and Technology, 48, 163-171.
• Georgieva, M., Nikolova, I., Bonchev, G., Katerova, Z., Todorova, D. 2015. A comparative analysis of membrane intactness and genome integrity in pea, barley, and wheat in response to UVC irradiation. Turkish Journal of Botany, 39, 1008-1013.
• Gong, M., Chen, B., Li, Z., Guo, L. 2001 Heat-shock-induced cross adaptation to heat, chilling, drought and salt stress in maize seedlings and involvement of H2O2. Journal of Plant Physiology, 158, 1125-1130.
• Gurmani, A.R., Bano, A., Khan, S.U., Din, J., Zhang, J.L. 2011. Alleviation of salt stress by seed treatment with abscisic acid (ABA), 6-benzylaminopurine (BA) and chlormequat chloride (CCC) optimizes ion and organic matter accumulation and increases yield of rice (Oryza sativa L.). Australian Journal of Crop Science, 5, 1278-1285.
• Hightower, K.R., McCready, J.P., Borchman, D. 1994. Membrane damage in UV-irradiated lenses. Photochemistry and Photobiology, 4, 485-490.
• Itoh, A., Kumashiro, T., Tanahashi, T., Nagakura, N., Nishi, T. 2002. Flavonoid glycosides from Rhazya orientalis. Journal of Natural Products, 65, 352 -357.
• Kalbina, I., Strid, A. 2006. The role of NADPH oxidase and MAP kinase phosphatase in UV-B-dependent gene expression in Arabidopsis. Plant, Cell and Environment, 29, 1783-1793.
• Kobashigawa, C., Tamaya, K., Shimomachi, T. 2011. Effect of UV-C treatment on plant growth and nutrient contents. Proc. 6th IS on Light in Horticulture, Eds.: E. Goto and S. Hikosaka, Acta Horticulturae, ISHS, 907.
• Ma, H., Xu, X., Feng, L. 2014. Responses of antioxidant defenses and membrane damage to drought stress in fruit bodies of Auricularia auricula-judae. World Journal of Microbiology and Biotechnology, 30, 119-124.
• Maciel, H.P.F., Gouvêa, C.M.C.P., Toyama, M., Smolka, M., Marangoni, S., Pastore, G.M. 2007. Extraction, purification and biochemical characterization of a peroxidase from Copaifera langsdorffii leaves. Química Nova, 30, 1067-1071
• . Manchenko, G.P. 2002. Handbook of Detection of Enzymes on Electrophoretic Gel, 1st ed., CRC Press, 163-165.
• Mohammadi, N., Mohammadi, S., Abdossi, V., Akbar Boojar, M.M. 2012. Effect of UV-C radiation on antioxidant enzymes in strawberry fruit (Fragaria x ananassa cv. Camarosa). ARPN Journal of Agricultural and Biological Science, 7, 860-864.
• Murashige, T., Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Plantarum, 15, 473-497
• . Navari-Izzo, F., Rascio, N. 1999. Plant response to water-deficit conditions. Chapter 10, (in) Handbook of Plant and Crop Stress 2nd ed., (eds.) Pessarakli, M., Marcel-Dekker Press, 231-271.
• Özen, F. 2002. Amsonia orientalis Decne (Apocynaceae). The Karaca Arboretum Magazine, 6, 169-172.
• Özen, F. 2006. Türkiye’de tükenme tehlikesinde olan bir türün otekolojisi: Amsonia orientalis Decne. (Apocynaceae). Balıkesir Üniversitesi Fen Bilimleri Enstitü Dergisi, 8, 4-9.
• Pan, Y., Wu, L.J., Yu, Z.L. 2006. Effect of salt and drought stress on antioxidant enzymes activities and SOD isoenzymes of liquorice (Glycyrrhiza uralensis Fisch). Plant Growth Regulation, 49, 157-165
• . Raghuvanshi, R., Sharma, R.K. 2016. Response of two cultivars of Phaseolus vulgaris L. (French beans) plants exposed to enhanced UV-B radiation under mountain ecosystem. Environmental Science and Pollution Research International, 23, 831-842.
• Rulcová, J., Pospíšilová, J. 2001. Effect of benzylaminopurine on rehydration of bean plants after water stress. Biologia Plantarum, 44, 75-81.
• Saxena, S.C., Joshi, P.K., Grimm, B., Arora, S. 2011. Alleviation of ultraviolet-C-induced oxidative damage through overexpression of cytosolic ascorbate peroxidase. Biologia, 66, 1052-1059.
• Severo, J., Tiecher, A., Pirrello, J., Regad, F., Latché, A., Pech, J-C., Bouzayen, M., Rombaldi, C.V. 2015. UV-C radiation modifies the ripening and accumulation of ethylene response factor (ERF) transcripts in tomato fruit. Postharvest Biology and Technology, 102, 9-16.
• Shiozaki, N., Hattori, I., Gajo, R., Tezuka, T. 1999. Activation of growth and nodulation in a symbiotic system between pea plants and leguminous bacteria by near UV radiation. Journal of Photochemistry and Photobiology B: Biology, 50, 33-37.
• Siddiqui, A., Dawar, S., Zaki, M.J., Hamid, N. 2011. Role of ultra violet (UV-C) radiation in the control of root infecting fungi on groundnut and mung bean. Pakistan Journal of Botany, 43, 2221-2224.
• Targovnik, A.M., Cascone, O., Miranda, M.V. 2012. Extractive purification of recombinant peroxidase isozyme c from insect larvae in aqueous two-phase systems. Separation and Purification Technology, 98, 199-205.
• Tevini, M. 2004. Plant responses to ultraviolet radiation stress. Chlorophyll a fluorescence: a signature of photosynthesis. G. C. Papageorgiou and Govindjee (Eds.). Springer, Netherlands, 605–621.
• Turan, S., Tripathy, B.C. 2013. Salt and genotype impact on antioxidative enzymes and lipid peroxidation in two rice cultivars during de-etiolation. Protoplasma, 250, 209-222.
• Valetti, N.W., Picó, G. 2013. A friendly method for Raphanus sativus L. (wild radish) peroxidase purification by polyelectrolyte precipitation. Seperation and Purification Technology, 119, 1-6.
• Younis, M.E.B., Hasaneen, M. N.A.G., Abdel-Aziz, H.M.M. 2010. An enhancing effect of visible light and UV radiation on phenolic compounds and various antioxidants in broad bean seedlings. Plant Signaling and Behavior, 5, 1197-1203.