Kiraz Anaçlarının in vitro Koşullarda Tuz Stresine Tolerans Mekanizmalarının Fizyolojik Parametreler ve Antioksidan Enzim İzoformları ile Belirlenmesi

Abstract

Tuzluluk, bitkilerde gelişim ve verimi sınırlayan önemli abiyotik stres koşullarının başında gelmektedir. Kiraz anaçlarının tuz stresi toleransları ile ilgili bilgiler oldukça sınırlıdır. Bu çalışmada, ülkemizde yetiştirilen üç kiraz anacının; Colt Prunus avium x Prunus pseudocerasus , Gisela 5 Prunus cerasus x Prunus avium ve Maxma Prunus mahaleb x Prunus avium , tuz stresine tolerans mekanizmaları in vitro koşullarda araştırılmıştır. Kiraz anaçlarının tuz stresine toleranslarını belirlemek amacıyla Murashige ve Skoog MS ortamına 0, 25 ve 50 mM NaCl uygulanmıştır. Anaçlarda; sürgün gelişimi, lipid peroksidasyonu MDA , membran geçirgenliği MG , toplam antioksidan aktivitesi TAA , prolin içeriği, toplam klorofil içeriği, katalaz CAT, EC 1.11.1.6 ve peroksidaz POD, EC 1.11.1.7 antioksidan enzim izoformları ile Na ve Cl konsantrasyonları belirlenmiştir. Tuz stresi anaçların sürgün gelişimini ve toplam klorofil içeriğini kontrole göre azaltırken MDA içeriğini, MG, TAA ve prolin içeriğini artırmıştır. Anaçların stres koşullarında sürgün POD enzim aktivitesi artmış ve üç farklı POD izoformu elde edilmiştir. Maxma anacının POD aktivitesi diğer anaçların POD aktivitesinden daha düşük olmuştur. Anaçların CAT izoformlarında ise uygulamalar ve anaçlar arasında belirgin bir farklılık elde edilmemiştir. Bütün parametreler birlikte değerlendirildiğinde, NaCl stresine Maxma anacının hassas, Colt anacının orta derecede hassas ve Gisela 5 anacının da dayanıklı olduğu belirlenmiştir

Keywords

• Enzim izoformu
• Katalaz
• Kiraz anaçları
• Peroksidaz
• Tuz stresi

Evaluation of Salt Tolerance Mechanisms with Physiological and Antioxidant Enzyme Isoform Parameters in in vitro Sweet Cherry Rootstocks

Abstract

Abiotic stress such as salinity is an important factor that limits plant growth and performance. Information regarding the genotypic variation for salinity stress tolerance in sweet cherry rootstocks is limited. In this study, salinity tolerance mechanisms of three sweet cherry rootstocks, namely; Colt Prunus avium x Prunus pseudocerasus , Gisela 5 Prunus cerasus x Prunus avium and Maxma Prunus mahaleb x Prunus avium , grown widely in Turkey were investigated under in vitro condition. The three rootstocks were cultured in vitro on MS medium supplemented with 0, 25 and 50 mM sodium chloride NaCl . The Shoot growth, lipid peroxidation MDA , membrane permeability MP , total antioxidant activity TAA , proline, total chlorophyll contents, catalase CAT, EC 1.11.1.6 and peroxidase POD, EC 1.11.1.7 antioxidant enzyme isoforms of rootstocks were studied. Compared to the control, salinity resulted in a reduction in the shoot growth and total chlorophyll contents. Contrary to this, MDA contents, MP, TAA and proline contents were increased by salinity. Activity of POD in the shoot of rootstocks was increased and three different POD isoforms were exhibited under saline conditions. The activity of POD was lower in the Maxma than the Colt and Gisela 5 rootstock. Salinity did not significantly change the CAT isoforms of the rootstocks. Regarding the all parameters studied, the rootstocks can be classified to their salt tolerance as sensitive Maxma , moderately sensitive Colt and resistant Gisela 5

Keywords

• Catalase
• Enzyme isoform
• Peroxidase
• Salinity stress
• Sweet cherry rootstocks

References

1. Alpaslan M & Gunes A (2001). Interactive effects of boron and salinity stress on the growth, membrane permeability and mineral composition of tomato and cucumber plants. Plant and Soil 236: 123-128
2. Arnon D M (1949). Copper enzymes in isolated chloroplasts polyphenoloxidase in Beta vulgaris. Plant Physiology 24: 1-15
3. Asish K P, Anath B D & Prasanna M (2004). Defense potentials to NaCl in mangrove, Bruguiera parviflora: Differential changes of isoforms of some antioxidative enzymes. Journal of Plant Physiology 161: 531-542
4. Aziz A, Martin-Tanguy J & Lather F (1999). Salt stress- induced proline accumulation and changes in tyramine and polyamine levels are linked to ionic adjustment in tomato leaf discs. Plant Science 145: 83-91
5. Balal R M, Ashraf M Y, Khan M M, Jaskani M J & Ashfaq M (2011). Influence of salt stress on growth and biochemical parameters of citrus rootstocks. Pakistan Journal of Botany 43(4): 2135-2141
6. Bates L S, Waldren R P & Teare I D (1973). Rapid determination of free proline for water-stress studies. Plant and Soil 39: 205-207
7. Bradford M M (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248-254
8. Cadenas S E (1989). Biochemistry of oxygen toxicity. Annual Review Biochemistry 58: 79-110

9. Cherian S & Reddy M P (2003). Evaluation of NaCl tolerance in the callus cultures of Suaeda nudiflora Moq. Biologia Plantarum 46: 193-198
10. Chinta S, Lakshmi A & Giridarakumar S (2001). Change in the antioxidant enzyme efficacy in two high yielding genotypes of mulberry (Morus alba L.) under NaCl salinity. Plant Science 161: 613-619
11. Cornic G (1996). Drought stress inhibits photosynthesis by decreasing stomatal aperture-not by affecting ATP synthesis. Trends in Plant Science 1: 21-26
12. Davies K J A (1987). Protein damage and degradation by oxygen radicals. I. General aspects. Journal of Biological Chemistry 262: 9895-9901
13. Dragišić Maksimović J, Zhang J, Zeng F, Živanović B D, Shabala L, Zhou M & Shabala S (2013). Linking oxidative and salinity stress tolerance in barley: Can root antioxidant enzyme activity be used as a measure of stress tolerance? Plant and Soil 365: 141-155
14. Eraslan F, Inal A, Savasturk O & Gunes A (2007). Changes in antioxidative system and membrane damage of lettuce in response to salinity and boron toxicity. Scientia Horticulturae 114: 5-10
15. Eraslan F, Inal A, Pilbeam D J & Gunes A (2008). Interactive effects of salicylic acid and silicon on oxidative damage and antioxidant activity in spinach (Spinacia oleracea L. cv. Matador) grown under boron toxicity and salinity. Plant Growth Regulation 55: 207-219
16. Erturk U, Sivritepe N, Yerlikaya C, Bor M, Ozdemir F & Turkan I (2007). Responses of the cherry rootstock to salinity in vitro. Biologia Plantarum 51(3): 597-600
17. FAO (2002). Crops and drops. Making the best use of water for agriculture. Food and Agriculture Organization of the United Nations, Rome, Italy
18. FAO (2012). Agricultural Statistics, Food and Agriculture Organization of the United Nations. http:\www. faostat.fao.org., Accessed: 29 October 2014
19. Fisarakis I, Chartzoulakis K & Stavrakas D (2001). Response of Sultana vines (V. vinifera L.) on six rootstocks to NaCl salinity exposure and recovery. Agricultural Water Management 51(1): 13-27
20. Fridovich I (1986). Biological effects of superoxide radical. Archives of Biochemistry Biophysics 247: 1-11
21. Gossett D R, Millhollon E P & Lucas M C (1994). Antioxidant response to NaCl stress in salt-tolerant and salt-sensitive cultivars of cotton. Crop Science 34: 706-714
22. Gulen H & Eris A (2004). Effect of heat stress on peroxidase activity and protein content in strawberry plants. Plant Science 166(3): 739-744
23. Güneş A, Çelik H, Alpaslan M, Söylemezoğlu G, Eraslan F, Yaşa Z & Koç Ö (2003). Asmaların (Vitis spp.) bor toksisitesi ve tuzluluğa karşı toleransının belirlenmesine yönelik olarak bor, sodyum ve klor alımlarının karşılaştırılması. Tarım Bilimleri Dergisi– Journal of Agricultural Sciences 9: 428-434
24. Gunes A, Inal A, Bagci E G & Pilbeam D (2007). Silicon-mediated changes of some physiological and enzymatic parameters symptomatic for oxidative stress in spinach and tomato grown in sodic-B toxic soil. Plant and Soil 290: 103-114
25. Hare P D & Cress W A (1997). Metabolic implication of stress-induced proline accumulation in plants. Plant Growth Regulation 21: 79-102
26. Heath R L & Packer L (1968). Photoperoxidation in isolated chloroplast. I. Kinetics and stoichiometry of fatty acid peroxidation. Archives of Biochemistry and Biophysics 125: 189-198
27. Imlay J A & Linn S (1988). DNA damage and oxygen radical toxicity. Science 240: 1302-1309
28. Ismail A M (2004). Response of maize and sorghum to excess boron and salinity. Biologia Plantarum 47(2): 313-316
29. Johnson J M & Ulrich A (1975). Analytical Methods for Use in Plant Analysis. II. California: Agricultural Experimental Station Bulletin
30. Karakullukçu E & Adak M S (2008). Bazı nohut (Cicer arietinum L.) çeşitlerinin tuza toleranslarının belirlenmesi. Tarım Bilimleri Dergisi-Journal of Agricultural Sciences 14(4): 313-319
31. Karimi H R & Hasanpour Z (2014). Effects of salinity and water stress on growth and macro nutrients concentration of pomegranate (Punica granatum L.). Journal of Plant Nutrition 37: 1937-1951
32. Kuşvuran Ş, Ellialtıoğlu Ş, Abak K & Yaşar F (2007). Bazı kavun (cucumis sp.) genotiplerinin tuz stresine tepkileri. Tarım Bilimleri Dergisi-Journal of Agricultural Sciences 13(4): 395-404
33. Kuşvuran Ş, Yaşar F, Abak K & Ellialtıoğlu Ş (2008). Tuz stresi altında yetiştirilen tuza tolerant ve duyarlı Cucumis sp.’nin bazı genotiplerinde lipid peroksidasyonu, klorofil ve iyon miktarlarında meydana gelen değişimler. Yüzüncü Yıl Üniversitesi, Ziraat Fakültesi, Tarım Bilimleri Dergisi 18(1): 13-20
34. Kotuby-Amacher J, Koening R & Kitchen B (2000). Salinity and Plant Tolerance. Electronic Publication AG-SO-03, Utah State University Extension, Logan.
35. Laemmli U K (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685
36. Lawlor D W (2001). Limitation of photosynthesis in water stressed leaves: Stomata vs. metabolism and the role of ATP. Annals of Botany 89: 1-15
37. Lee D H, Kim T S & Lee C B (2001). The inductive responses of the antioxidant enzymes by salt stress in the rice (Oryza sativa L.). Journal of Plant Physiology 158: 737-745
38. Liebler D C, Kling D S & Reed D J (1986). Antioxidant protection of phospholipid bilayers by α-tocopherol. Control of α-tocopherol status and lipid peroxidation by ascorbic acid and glutathione. Journal of Biological Chemistry 261: 12114-12119
39. Molassiotis A N, Sotiropoulos T E, Tanou G, Kofidis G, Diamantidis G & Therios I (2006). Antioxidant and anatomical responses in shoot culture of the apple rootstock MM 106 treated with NaCl, KCl, mannitol or sorbitol. Biologia Plantarum 50(1): 61-68
40. Murashige T & Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15(3): 473-497
41. Neocleous D & Vasilakakis M (2007). Effects of NaCl stress on red raspberry (Rubus idaeus L. ‘Autumn Bliss’). Scientia Horticulturae 112: 282-289
42. Oren R, Sperry J S, Katul G G, Pataki D E, Ewers B E, Phillips N & Schaffer K V R (1999). Survey and synthesis of intra and inter specific variation of stomatal sensitivity to vapour pressure deficit. Plant Cell & Environment 22: 1515-1526
43. Papadakis I E, Veneti G, Chatzissavvidis C, Sotiropoulos T E, Dimassi K N & Therios I N (2007).Growth, mineral composition, leaf chlorophyll and water relationships of two cherry varieties under NaCl- induced salinity stress. Soil Science & Plant Nutrition 53(3): 252-258
44. Prieto P, Pineda M & Aguilar M (1999). Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: Specific application to the determination of vitamin E. Analytical Biochemistry 269: 337-341
45. Ruiz D, Martinez V & Cerda A (1999). Demarcating specific ion (NaCl, Cl-, Na+) and osmotic effects in the response of two citrus rootstocks to salinity. Scientia Horticulturae 80: 213-224
46. Sairam R K & Saxena D C (2000). Oxidative stress and antioxidants in wheat genotypes: Possible mechanism of water stress tolerance. Journal of Agronomy & Crop Science 184: 55-61
47. Sairam R K, Deshmukhi P S & Saxena D C (1998). Role of antioxidant systems in wheat genotypes tolerance to water stress. Biologia Plantarum 41(3): 387-394
48. Sekmen A H, Turkan I & Takio S (2007). Differential responses of antioxidative enzymes and lipid peroxidation to salt stress in salt-tolerant Plantago maritima and salt-sensitive Palantago media. Physiologia Plantarum 131(3): 399-411
49. Shalata A & Tal M (1998). The effect of salt stress on lipid peroxidation and antioxidants in the leaf of the cultivated tomato and its wild salt-tolerant relative Lycopersicon pennellii. Physiologia Plantarum 104: 169-174
50. Shen H & Yan X (2002). Membrane permeability in roots of crotalaria seedlings as affected by low temperature and low phosphorus stress. Journal of Plant Nutrition 25(5): 1033-1047
51. Singh S K, Sharma H C, Goswami A M, Datta S P & Singh S P (2000). In vitro growth and leaf composition of grapevine cultivars as affected by sodium chloride. Biologia Plantarum 43(2): 283-286
52. Sotiropoulos T E, Dimassi K N, Tsirakoglou V & Therios I N (2006a). Responses of two Prunus rootstocks to KCl induced salinity in vitro. Biologia Plantarum 50(3): 477-480
53. Sotiropoulos T E, Therios I N, Almaliotis D, Papadakis I & Dimassi K N (2006b). Response of cherry rootstocks to boron and salinity. Journal of Plant Nutrition 29: 1691-1698
54. Sotiropoulos T E (2007). Effect of NaCl and CaCl2 on growth and contents of minerals, chlorophyll, proline and sugars in the apple rootstock M 4 cultured in vitro. Biologia Plantarum 51(1): 177-180
55. Tarakcioglu C & Inal A (2002). Changes induced by salinity, demarcating specific ion ratio (Na/Cl) and osmolality in ion and proline accumulation, nitrate reductase activity and growth performance of lettuce. Journal of Plant Nutrition 25(1): 27-41
56. Termaat A & Munns R (1986). Use of concentrated macronutrient solutions to separate osmotic from NaCl-specific effects on plant growth. Australian Journal of Plant Physiology 13: 509-522
57. Troncoso A, Matte C, Cantos M & Lavee S (1999). Evaluation of salt tolerance of in vitro-grown grapevine rootstock varieties. Vitis 38: 55-60
58. Turner N C, Colmer T D, Quealy J, Pushpavalli R, Krishnamurthy L, Kaur J, Singh G, Siddique K H M & Vadez V (2013). Salinity tolerance and ion accumulation in chickpea (Cicer arietinum L.) subjected to salt stress. Plant and Soil 365: 347-361
59. Wang X & Han J (2009). Changes of proline content, activity, and active isoforms of antioxidative enzymes in two alfalfa cultivars under salt stress. Agricultural Sciences in China 8(4): 432-440
60. Wendel J F & Weeden N F (1989). Visualization and interpretation of plant isozymes. In: D E Soltis & P S Soltis (Eds), Isozymes in Plant Biology, Dioscorides Press, Portland, Oregon, pp. 5-44
61. Withan F H, Blaydes D F & Dewlin R M (1971). Experiments in Plant Physiology. New York, Von Nostrand Reinhold Company, pp. 55-56
62. Wolfe W H (1976). Identification of grape varieties by isozyme banding patterns. American Journal of Enology Viticulture 27: 68-73
63. Yaşar F, Ellialtıoğlu Ş, Özpay T & Uzal Ö (2008). Tuz stresinin karpuzda (Citrullus lanatus (Thunb.) Mansf.) antioksidatif enzim (SOD, CAT, APX ve GR) aktivitesi üzerine etkisi. Yüzüncü Yıl Üniversitesi, Ziraat Fakültesi, Tarım Bilimleri Dergisi 18(1): 51-55