Domates Bitkisinde Kurşun Pb ve Kadminyumun Cd Lipid Peroksidasyonu, Katalaz CAT Enzim Aktivitesi ve Gen Ekpresyon Profiline Etkisi

Abstract

Önemli bir abiyotik stres faktörü olan ağır metaller bitkilerde çok çeşitli yanıt mekanizmalarını uyarabilirler. Bu yanıt mekanizmaları; membran kompozisyonunda değişiklik, küçük molekül ve serbest radikallerin üretimi, antioksidant enzimlerin aktivitelerinin ya da gen ekspresyonlarının değişimini içerir. Bu sebeple, bu araştırmada çeşitli konsantrasyonlardaki 0, 80, 160, 320, 640 ve 1280 μM Pb2+ve Cd2+ kontaminasyonuna maruz kalan domates bitkilerinde lipid peroksidasyon seviyesi MDA , katalaz enzim aktivitesi ve real-time PCR aracılığı ile katalaz gen ekpresyon seviyesi belirlenmiştir. 320 ve 640 μM Cd+2 kontaminasyonu hariç tüm Cd+2 ve Pb+2 kontaminasyonları lipid peroksidasyonuna ve katalaz enzim aktivitesinde artışa neden olmuştur. Sonuç olarak; çeşitli konsantrasyonlarda Pb2+ ve Cd2+ kontaminasyonuna maruz kalan domates bitkisinde, CAT gen ekpresyonu ve lipid peroksidayonu arasında pozitif korelasyon bulunurken, CAT gen ekpresyonu ve enzim aktivitesi arasında korelasyon tespit edilememiştir. Bu durum genlerin transkripsiyonel, posttranskripsiyonel ve aynı zamanda translasyonel veya posttranslasyonel seviyelerdeki regülasyonu ile açıklanabilir

Keywords

• Domates Solanum lycopersicum L.
• Lipid peroksidasyonu
• CAT enzim aktivitesi
• Gen ekpresyonu
• Real-time PCR

Effects of Lead Pb and Cadmium Cd Elements on Lipid Peroxidation, Catalase Enzyme Activity and Catalase Gene Expression Profile in Tomato Plants

Abstract

Heavy metals are significant abiotic stress factor, affecting various response mechanisms in plants. These responses include: changes in membrane composition, production of small molecules and free radicals, and alterations in the activities of antioxidant enzymes and their gene expressions. For this reason, lipid peroxidation levels MDA , catalase enzyme activity, and gene expression profiles, quantified by real-time PCR, were analyzed in tomato plants exposed to various concentrations 0, 80, 160, 320, 640 and 1280 μM of Cd2+ and Pb2+. All concentration of Cd+2 or Pb+2 contamination led to increased lipid peroxidation and catalase enzyme activity, except for 320 and 640 μM Cd+2 contamination levels. As a result, gene expression patterns at the mRNA level and changes in MDA content under different concentrations of Pb+2 and Cd+2 contamination revealed a positive correlation, although no correlation was found between gene expression patterns at the mRNA level and catalase enzyme activity. These results might be explained by the regulation of genes at the transcriptional, posttranscriptional, and also translational or posttranslational levels

Keywords

• Tomato Solanum lycopersicum L.
• Lipid peroxidation
• CAT enzyme activity
• Gene expression
• qRT-PCR

References

1. Aebi H (1984). Catalase in vitro. Methods in Enzymology 105: 121-126
2. Asada K & Takahashi M (1987). Production and scavenging of active oxygen in photosynthesis. In: Kyle DJ, Osmond CJ, Artzen CJ, editors. Photoinhibition: Topics in Photosynthesis. Amsterdam, Elsevier, pp. 227-287
3. Ben Ammar W, Nouairi I, Tray B, Zarrouk M, Jemal F & Ghorbel M H (2005). Cadmium effects on mineral nutrition and lipid contents in tomato leaves (in French). Journal of Social Biology 199: 157-163
4. Cervilla L M, Blasco B, Rios J J, Romero L & Ruiz J M (2007). Oxidative stress and antioxidants in tomato (Solanum lycopersicum) plants subjected to boron toxicity. Annals of Botany 100: 747-756
5. Cui L, Lee H S, Ndinteh D T, Mbafor J T, Kim Y H, Tv L E, Nguyen P H & Wk O H (2010). New prenylated flavanones from Erythrina abyssinica with protein tyrosine phosphatase 1B (PTP1B) inhibitory activity. Planta Medica 76: 713-718
6. El-Beltagi H S, Mohamed A A & Rashed M M (2010). Response of antioxidative enzymes to cadmium stress in leaves and roots of radish (Raphanus sativus L.). Notulae Scientia Biologicae 2: 76-82
7. Farrel R E (2007). The regulation of gene expression in plants and animals. In: Bassett CL, editor. Regulation of Gene Expression in Plants: The Role of Transcript Structure and Processing. New York, NY: Springer, pp. 1-34
8. Fortunato A, Lidon F C, Batista-Santos P, Leitão A E, Pais I P, Ribeiro A I & Ramalho J C (2010). Biochemical and molecular characterization of the antioxidative system of Coffea sp. under cold conditions in genotypes with contrasting tolerance. Journal of Plant Physiology 167: 333-342

9. Gaur A & Grupa S K (1994). Lipid components of mustard seeds (Brassica juncea L.) as influenced by cadmium levels. Plant Food for Human Nutrition 46: 93-102
10. Goupil P, Souguir D, Ferjani E, Faure O, Hitmi A & Ledoigt G (2009). Expression of stress-related genes in tomato plants exposed to arsenic and chromium in nutrient solution. Journal of Plant Physiology 166: 1446-1452
11. Gratao P L, Polle A, Lea P J & Azevedo R A (2005). Making the life of heavy metal-stressed plants a little easier. Functional Plant Biology 32: 481-494
12. Henriques F (2003). Gas exchange, chlorophyll a fluorescence kinetics and lipid peroxidation of pecan leaves with varying manganese concentrations. Plant Science 165: 239-244
13. Hernandez J A, Olmos E, Corpas F J, Sevilla F & De1 Rio L A (1995). Salt-induced oxidative stress in chloroplasts of pea plants. Plant Science 105: 151-167
14. Hodges D M, Delong J M, Forney C F & Prange R K (1999). Improving the thiobarbituric acid-reactive- substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta 207: 604-611
15. Jovanovic Z, Milosevic J & Radovic S (2006). Antioxidative enzymes in the response of buckwheat (Fagopyrum esculentum Moench) to ultraviolet B radiation. Journal of Agriculture and Food Chemistry 54: 9472-9478
16. Kabata-Pendias A & Pendias H (1989). Trace Elements in the Soil and Plants. Boca Raton, CRC Pres
17. Krupa Z & Baszynski T (1985). Effects of cadmium on the acyl lipid content and fatty acid composition in thylakoid membranes isolated from tomato leaves. Acta Physiology Plantarum 7: 55-64
18. Krupa Z & Baszynski T (1989). Acyl lipid composition of thylakoid membranes of cadmium-treated tomato plants. Acta Physiology Plantarum 11: 111-116
19. Lake D L, Kirk P W W & Lester J N (1984). Fractionation, characterization, and speciation of Heavy metals in sewage and sludge amended soil: A review. Jornal of Environmental Quality 13: 175-183
20. Lee S H & An C S (2005). Differential expression of three catalase genes in hot pepper (Capsicum annuum L.). Molecules and Cells 20: 247-255
21. Lima L, Seabra A, Melo P, Cullimore J & Carvalho H (2006). Phosphorylation and subsequent interaction with 14-3-3 protein regulates plastid glutamine synthetase in Medicago truncatula. Planta 223: 558-567
22. Livak K J & Schmittgen T D (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods 25: 402- 408
23. Lyons J M (1973). Chilling injury in plants. Annual Review of Plant Physiology 24: 445-466
24. Malik D, Sheoran I S & Singh R (1992). Lipid composition of thylakoid membranes of cadmium treated wheat seedlings. Indian Journal of Biochemistry & Biophysics 29: 350-354
25. Matsumura H, Xie Y, Shirakata S, Inoue T, Yoshinaga T, Ueno Y, Izui K & Kai Y (2002). Crystal structures of C4 form maize and quaternary complex of E. coli phosphoenolpyruvate carboxylases. Structure 10: 1721-1730
26. Mead J F (1976). Free radical mechanisms of lipid damage and consequences for cellular membranes. In: WA Pryor, editor. Free Radicals in Biology, Vol 1. New York, Academic Press, pp. 51-68
27. Mittler R (2002). Oxidative stress, antioxidants and stres tolerance. Trends in Plant Science 7: 405-410
28. Mittler R, Vanderauwera S, Gollery M & Van Breusegem F (2004). The reactive oxygen gene network in plants. Trends in Plant Science 9: 490-498
29. Mittova V, Theodoulou F L, Kiddle G, Gomez L, Volokita M, Tal M, Foyer C H & Guy M (2003). Coordinate induction of glutathione biosynthesis and glutathione- metabolizing enzymes is correlated with salt tolerance in tomato. FEBS Letter 554: 417-421
30. NCBI (2013a). Nucleotide. http://www.ncbi.nlm.nih.gov/ nuccore/M93719.1 (Erişim tarihi: 10.01.2014)
31. NCBI (2013b). Nucleotide http://www.ncbi.nlm.nih. gov/nuccore/NM_001308447.1 17.01.2014) (Erişim tarihi
32. Nouairi I, Ben Ammar W, Ben Youssef N, Ben Mileddaoud D, Habibghorbal M & Zarrouk M (2006). Comparative study of cadmium effects on membrane lipid composition of Brassica juncea and Brassica napus leaves. Plant Science 170: 511-519
33. Polle A, Chakrabarti K, Chakrabarti S, Seifert F, Schramel P & Rennenberg H (1992). Antioxidants and manganese deficiency in needles of Norway spruce (Picea abies L.) trees. Plant Physiology 99: 1084-1089
34. Qilin D, Jin W, Bin F, Tingting L, Chen C, Honghui L & Shizhang D (2009). Molecular cloning and characterization of a new peroxidase gene (OvRCI) from Orychophragmus violaceus. African Journal of Biotechnology 8(23): 6511-6517
35. Quariti O, Boussama N, Zarrouk M, Cherif A & Ghorbalm H (1997). Cadmium and copper-induced changes in tomato membrane lipids. Phytochemistry 45: 1343- 1350
36. Rao A Q, Hussain S S, Shahzad M S, Bokhari Sy A, Raza M H, Rakha A, Majeed A, Shahid A A, Saleem Z, Husnain T & Riazuddin S (2006). Somatic embryogenesis in wild relatives of cotton (Gossypium spp.). Journal of Zhejiang University Science B 7(3): 1673-1581
37. Schmidt M, Dehne S & Feierabend J (2002). Post- transcriptional mechanisms control catalase synthesis during its light-induced turnover in rye leaves through the availability of the hemin cofactor and reversible changes of the translation efficiency of mRNA. Plant Journal 31: 601-613
38. Scott D, Keoghan J M & Allen B E (1996). Native and low input grasses-a New Zealand high country perspective. New Zealand Journal of Agricultural Research 39: 499-512
39. Sharma R K, Agrawal M & Marshall F (2006). Heavy metals contamination in vegetables grown in wastewater irrigated areas of Varanasi, India. Bulletin of Environmental Contamination and Toxicology 77: 312-318
40. Sharma R K, Agrawal M & Marshall F M (2007). Heavy metals contamination of soil and vegetables in suburban areas of Varanasi, India. Ecotoxicology and Environmental Safety 66: 258-266
41. Shim I S, Momose Y, Yamamoto A, Kim D W & Usui K (2003). Inhibition of catalase activity by oxidative stress and its relationship to salicylic accumulation in plants. Plant Growth Regulation 39: 285-292
42. Sinha S, Saxena R & Singh S (2005). Chromium induced lipid peroxidation in the plants of Pistias tratiotes L., role of antioxidants and antioxidant enzymes. Chemosphere 58: 595-604
43. Skorzynska-Polit E, Drazkiewicz M & Krupa Z (2010). Lipid peroxidation and antioxidative response in Arabidopsis thaliana exposed to cadmium and copper. Acta Physiology Plantarum 32: 169-175
44. Smirnoff N (1993). The role of active oxygen in the response of plants to water deficit and desiccation. New Phytology 125: 27-58
45. Soydam Aydın S, Büyük İ, Çelikkol B P & Aras S (2013). A role of catalase (CAT) in detoxification of reactive oxygen species (ROS) in tomato (Lycopersicum esculentum) contaminated with manganese (Mn2+). Biological Diversity and Conservation 6: 140-145
46. Trichopoulos D (1997). Epidemiology of Cancer. In: DeVita VT editor. Cancer: Principles and Practice of Oncology, Philadelphia, Lippincott Company, pp. 231-258
47. Türkdoğan M K, Kilicel F, Kara K, Tuncer I & Uygan I (2002). Heavy metals in soil, vegetables and fruits in the endemic upper gastrointestinal cancer region of Turkey. Environmental Toxicology and Pharmacology 13: 175-179
48. Vassilev A (2004). Cadmium-induced changes in chloroplast lipids and photosystem activities in barley plants. Biologia Plantarum 48: 153-156
49. Voutsa D A, Grimanis A & Samara C (1996). Trace elements in vegetables grown in industrial areas in relation to soil and air particulate matter environ. Environmental Pollution 94: 325-335
50. Wang D, Heckathorn S A, Barua D, Joshi P, Hamilton E W & Croix J L A (2008). Effects of elevated CO2 on the tolerance of photosynthesis to acute heat stress in C, C and CAM species. American Journal of Botany 3 4 95: 165-176