Effect of Cadmium on changes of some enzymatic and none-enzymatic antioxidant defense systems in lentil seedlings (Lens culinaris Medik.)

پژوهش‌های حبوبات ایران

Article 8, Volume 7, Issue 2, December 2017, Pages 125-137   

Document Type: Original Articles

DOI: 10.22067/ijpr.v7i2.45542

Authors

Hamid Reza Kavousi^{* 1}; Fatemeh Barandeh²

¹Shahid Bahonar University

²Shahid Bahonar University of Kerman

Abstract

Introduction
Throughout their life cycle, plants are subjected to various types of environmental stresses which include salinity, water deficit, temperature extremes, toxic metal ion concentration and UV radiation. These environmental factors limit the growth and productivity of plants to various degrees, depending upon severity of stress. Heavy metal toxicity is one of the major current environmental health problems and potential dangerous due to bioaccumulation through the food chain and in plant products for human consumption. Cadmium (Cd) is a high toxic trace element that enters the environment mainly from industrial processes and phosphate fertilizers. It can reach high levels in agricultural soil and is easily accumulated in plants. Cd ions are taken up readily by the plant roots and translocated to the above-ground vegetative parts. The presence of Cd at higher concentrations in the soil damages root tips, reduces nutrient and water uptake, impairs photosynthesis and inhibits growth of the plants. Furthermore, Cd directly or indirectly induces reactive oxygen species (ROS), which affect the redox status of the cell and cause oxidative damage to proteins, lipids, and other biomolecules. Cd damages the nucleoli in cells of the root tip, alters the synthesis of RNA, inhibits ribonuclease activity and inhibits the DNA repairing mechanism. Understanding the biochemical and molecular responses to Cd stress is essential for a holistic opinion of plant resistance mechanisms of heavy metal stress.

Materials and Methods
In this study, the effect of different concentrations of cadmium on proline and phenolic contents, activity of some antioxidative enzymes such as superoxide dismutase, catalase and ascorbate peroxidase, phenylalanine ammonialyase in Lense culinaris Medik. was studied. Therefore, two-week-old plantlets were treated with different concentrations of cadmium chloride (0 as control, 0.125, 0.25, 0.5, 1, 2.5 and 5 mM) of cadmium chloride for 10 days and then the above mentioned factors were investigated. The experiments were carried out by using a complete randomized block design with three replications. The statistical analyses were carried out using the SAS version 9. Changes in biochemical parameters were tested statistically by performing one-way analysis of variance (ANOVA). The treatment means separated using Duncan's multiple-range test (DMRT) taking P < 0.05 as significant.

Results and Discussion
Surveying the results indicated that the amount of non-enzymatic antioxidant compounds (phenol and proline) were increased in treated seedlings compared with the control. Review of the literature indicates that a stressful environment results in an overproduction of proline in plants which in turn imparts stress tolerance by maintaining cell turgor or osmotic balance; stabilizing membranes thereby preventing electrolyte leakage; and bringing concentrations of ROS within normal ranges, thus preventing oxidative burst in plants. The increase in phenolic content may be due to protective function of these compounds against heavy metal stress by metal chelation and ROS scavenging. The results obtained from measuring the activity of antioxidant enzymes showed that with increasing concentrations of cadmium, the activity of antioxidant enzymes superoxide dismutase, catalase and ascorbate peroxidase were increased in lentil seedlings. However, this increase was more evident at concentrations greater than 0.5 mM. These results suggest that lentil seedlings tend to cope with free radicals generated by Cd through coordinated, enhanced activities of the antioxidative enzymes involved in detoxification. With increasing levels of cadmium in the medium, the activity of the phenylalanine ammonialyase also increased. The trend of phenylalanine ammonialyase activity induction in response to different concentrations of cadmium, was similar to those of antioxidant enzymes. Although the induction of activity of this enzyme was higher than the antioxidant enzymes. PAL is a key enzyme in the phenolic metabolism that has been reported to protect plants against stress conditions via synthesizing various phenylpropanoid products such as simple phenols, anthocyanin, flavonoid, and lignin.

Conclusion
The present study demonstrated that antioxidative system in L. culinaris underwent biochemical changes to survive under high concentrations of cadmium. Increase in metal chelate components (free phenols and proline) in all treatment levels proves this fact. Proline and phenolic compounds produced by the Phenylpropanoid Pathway in lentil played a major role in its response to Cd stress. Also that prolonged stress induced by Cd concentrations, can result into the activation of antioxidant enzymes such as superoxide dismutase, catalase and ascorbate peroxidase.

Key words: Antioxidant enzymes, Cadmium, Lentil, Phenylalanine ammonialyase, Total phenolics

Keywords

Antioxidant enzymes, Cadmium, Lentil, Phenylalanine ammonialyase; Total phenolics

References

1. Ahmadvand, S., Bahmani, R., Habibi, D., and Forouzesh, P. 2013. Investigation of cadmium chloride effect on growth parameters and some physiological characteristics in bean (Phaseolus vulgaris L.) seedlings. Journal of Agronomy and Plant Breeding 8(4): 167-182. (In Persian).

2. Babula, P., Adam, V., Opatrilova, R., Zehnalek, J., Havel, L., and Kizek, R. 2008. Uncommon heavy metals, metalloids and their plant toxicity: a review. Environmental Chemistry Letters 6: 189-213.

3. Badpa, K., Movahhedi Dehanvi, M., and Yadavi, A. 2015. Interaction of cadmium nitrite and salicylic aside on antioxidant enzyme activity and leaf soluble protein and malondialdehyde of safflower (Carthamus tinctorius L. cv. Soffe). Journal of Plant Process and Function 3(9): 21-32. (In Persian with English Summary).

4. Bates, L.S., Waldren, R.P., and Teare, I.D. 1973. Rapid determination of free proline for water-stress studies. Plant and Soil 39: 205-207.

5. Beers, R.F., and Sizer, I.W. 1952. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. Journal of Biological Chemistry 195: 133-140.

6. Benavides, M.P., Gallego, S.M., and Tomaro, M.L. 2005. Cadmium toxicity in plants. Brazilian Journal of Plant Physiology 17: 21-34.

7. Boudet, A.M. 2007. Evolution and current status of research in phenolic compounds. Phytochemistry 68: 2722-2735.

8. Bradford, M.M. 1976. A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248-254.

9. Callahan, D.L., Baker, A.J.M., Kolev, S.D., and Wedd, A.G. 2005. Metal ion ligands in hyper accumulating plants. Journal of Biological Inorganic Chemistry 11: 2-12.

10. Chu, Y.H., Chang, C.L., and Hsu, H.F. 2000. Flavonoid contents of several vegetable and their antioxidant activity. Journal of Agriculture and Food Science 80: 561-566.

11. Dhindsa, R.A., Plumb-Dhindsa, P., and Thorpe T.A. 1981. Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation, and decreased levels of superoxide dismutase and catalase. Journal of Experimental Botany 126: 93-101.

12. Dinakar, N., Nagajyothi, P.C., Suresh, S., Damodharam, T., and Suresh, C. 2009. Cadmium induced changes on proline, antioxidant enzymes, nitrate and nitrite reductases in Arachis hypogaea L. Journal of Environmental Biology 30(2): 289-294.

13. Dudka, S., Piotrowska, M., and Terelak, H. 1996. Transfer of cadmium, lead and zinc from industrially contaminated soil to crop plants: A field study. Environmental Pollution 94: 181-188.

14. Ferreira, R.R., Fornazir, R.F., Vitoria, A.P., and Lea, P.J. 2002. Changes in antioxidant enzyme activities in soybean under cadmium stress. Journal of plant Nutrition 25: 327-342.

15. Hayat, S., Hayat, Q., Alyemeni, M.N., Wani, A.S., Pichtel, J., and Ahmad, A. 2012. Role of proline under changing environments. Plant Signaling and Behavior 7(11): 1456-1466.

16. John, R., Ahmad, P., Gadgil, K., and Sharma, S. 2008. Effect of cadmium and lead on growth, biochemical parameters and uptake in Lemna polyrrhiza L. Plant, Soil & Environment 54(6): 262-270.

17. Khatun, S., Babar Ali, M., Hahn, E.J., and Paek, K.Y. 2008. Copper toxicity in Withania somnifera: Growth and antioxidant enzymes responses of in vitro grown plants. Environmental and Experimental Botany 64: 279-285.

18. Kovacik, J., and Backor, M. 2007. Phenylalanine ammonialyase and phenolic compounds in chamomile tolerance to cadmium and copper excess. Water, Air & Soil Pollution 185: 185-193.

19. Kliebenstein, D.J. 2004. Secondary metabolites and plant/environment interactions: a view through Arabidopsis thaliana tinged glasses. Plant Cell and Environment 27: 675-684.

20. Laspina, N.V., Groppa, M.D., Tomaro, M.L., and Benavides, M.P. 2005. Nitric oxide protects sunflower leaves against Cd-induced oxidative stress. Plant Sciences 169(2): 323-330.

21. Mohammadi, M., Habibi, D., Ardakani, M., and Asgharzadeh, A. 2011. Effect of biologic fertilizers, homic acid and super absorbent polymer on chlorophyll content, membrane and superoxide dismutase and catalase activity in annual Medicago species under cadmium toxicity. 6(2): 79-65. (In Persian).

22. Mittler, R. 2002. Oxidative stress, antioxidants and stress tolerance. TRENDS in Plant Science 7(9): 405-410.

23. Myung-Min, H., Trick, H.N., and Rajasheka, E.B. 2009. Secondary metabolism and antioxidant are involved in environmental adaptation and stress tolerance in lettuce. Journal of Plant Physiology 166: 180-191.

24. Nakano, Y., and Asada, K. 1987. Purification of ascorbate peroxidase in spinach chloroplast: in inactivation in ascorbate-depleted medium and reactivation by monodehydroascorbate radical. Plant Cell Physiology 28: 131-140.

25. Pandey, P., and Tripathi, A.K. 2011. Effect of heavy metals on morphological and biochemical characteristics of Albizia procera benth seedlings. International Journal of Environmental Sciences 1(5): 1009-1018.

26. Rastgoo, L., and Alemzadeh, A. 2011. Biochemical responses of Gouan (Aeluropus littoralis) to heavy metals stress. Australian Journal of Crop Science 5(4): 375-383.

27. Schutzendubel, A., Schwanz, P., Teichmann, T., Gross, K., Langenfeld, R., Douglas, L., and Polle, A. 2001. Cadmium-induced changes in antioxidative systems, hydrogen peroxide content, and differentiation in Scots pine roots. Plant Physiology 127: 887-898.

28. Shahid, M., Pourrut, B., Dumat, C., Nadeem, M., Aslam, M., and Pinelli, E. 2014. Heavy-metal induced reactive oxygen species: phytotoxicity and physicochemical changes in plants. Reviews of Environmental Contamination and Toxicology 232: 1-44.

29. Solecka, D. 1997. Role of phenylpropanoid compounds in plant responses to different stress factor. Plant Physiology 19: 257-268.

30. Sun, R.L., Zhou, Q.X., Sun, F.H., and Jin, C.X. 2007. Antioxidative defense and proline/phytochelatin accumulation in a newly discovered Cd hyperaccumulator, Solanum nigrum L. Environmental and Experimental Botany 60: 468-476.

31. Tewari, R.K., Kumar, P., and Sharma, P.N. 2008. Morphology and physiology of zinc-stressed mulberry plants. Journal of Plant Nutrition and Soil Science 171: 286-294.

32. Tian, X., and Li, Y. 2007. Nitric oxide treatment alleviates drought stress in wheat seedlings. Biologia Plantarum 50: 775-778.

33. Torres, E., Cid, A., Herrero, C., and Abalde, J. 2000. Effect of cadmium on growth, ATP content, carbon fixation and ultra-structure in the marine diatom Phaeodactylum tricornutum Bohlin. Water, Air & Soil Pollution 117: 1-14.

34. Unyayar, S., Kele, Y., and Cekic, F.O. 2005. The antioxidative response of two tomato species with different drought tolerances as a result of drought and cadmium stress combinations. Plant Soil Environment 51(2): 57-64.

35. Vassilev, A., Tsonev, T., and Yordanov, I. 1998. Physiological response of barley plants (Hordeum vulgare) to cadmium contamination in soil during ontogenesis. Environmental Pollution 103: 287-293.

36. Velioglu, Y.S., Mazza, G., Gao, L., and Oomah, B.D. 1998. Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products. Journal of Agricultural and Food Chemistry 46: 4113-4117.

37. Vogt, T. 2010. Phenylpropanoid biosyntesis. Molecular Plant 3: 2-20.

38. Wang, J.W., Zheng, L.P., Wu, J.Y., and Tan, R.X. 2006. Involvement of nitric oxide in oxidative burst phenylalanine ammonialyase activation and taxol production induced by low-energy ultrasound in Taxus yunnanensis cell suspension cultures. Nitric Oxide Biology and Chemistry 15: 351-358.

39. Zhang. F., Shi, W., Jim, Z., and Shen, Z. 2003. Response of antioxidative enzymes in cucumber chloroplasts to cadmium toxicity. Journal of Plant Nutrition 26: 1779-1788.

40. Zhang, X., Fan, X., Li, C., and Nan, Z. 2010. Effects of cadmium stress on seed germination, seedling growth and antioxidative enzymes in Achnatherum inebrians plants infected with a Neotyphodium endophyte. Plant Growth Regulation 60: 91-97.