تأثیر دمای شبانه و نیتریک‌اکساید بر برخی ویژگی های فیزیو-بیوشیمیایی گیاه نخود

نوع مقاله : مقالات پژوهشی

نویسندگان

دانشگاه شهید مدنی آذربایجان

چکیده

تنش های محیطی از جمله دماهای پایین باعث کاهش تولید و کیفیت محصولات زراعی می شوند. نیتریک‌اکساید به‌عنوان یکی از تنظیم‌کننده های رشد گیاهی نقش مهمی در کاهش اثرات سوء ناشی از تنش ها برعهده دارد. در این پژوهش تغییرات صفات فیزیو-بیوشیمیایی گیاه نخود تحت تیمار نیتریک‌اکساید و دمای شبانه مورد‌بررسی قرار گرفت. وزن‌تَر بخش های هوایی و ریشه‌ها و ترکیباتی مانند پرولین، قندهای محلول و نامحلول، آسکوربات و فعالیت آنزیم های آسکوربات‌پراکسیداز و پلی فنل اکسیداز سنجش شدند. نتایج نشان‌داد که اثر نیتریک‌اکساید بر وزن‌تَر بخش های هوایی و محتوای پرولین، قندهای محلول و فعالیت آنزیم پلی فنل اکسیداز معنی-دار بود. تغییر دما بر وزن‌تَر ریشه، محتوای پرولین، قندهای محلول و نامحلول، آسکوربات، فعالیت آنزیم‌های آسکوربات پراکسیداز و پلی فنل اکسیداز تأثیر معنی دار داشت. برهم کنش نیتریک‌اکساید و دما بر وزن‌تَر ریشه و بخش های هوایی، محتوای پرولین، قندهای محلول و فعالیت آنزیم آسکوربات‌پراکسیداز تأثیر معنی داری را نشان داد. چنین ارزیابی می‌شود که متابولیسم نخود به دمای شبانه وابسته‌ بوده و تیمار نیتریک‌اکساید الگوی تغییرات متابولیکی را تحت تأثیر قرار می‌دهد.

کلیدواژه‌ها


عنوان مقاله [English]

The effects of night temperature and nitric oxide on some physio-biochemical characters of Pea plants

نویسندگان [English]

  • Nader Chaparzadeh
  • Masumeh Faraji
Azarbaijan Shahid Madani University
چکیده [English]

Introduction
Plant responses to environmental stress have a central role in agricultural production. Responses results from events occurring at all levels of the organization, from biochemical reactions in cells to whole plant physiology. Many plants are injured when exposed to low non-freezing temperatures. However, data on the effects of night temperatures are scarce. On the other hand, nitric oxide (NO), as a plant growth regulator, has an important role in ameliorating stress induced damage in plants. Therefore, the aim of this work was to evaluate the role of NO during low temperature nights. In this research the changes in some physio-biochemical characters of Pea plants under NO and night temperature treatments were studied.

Materials and Methods
The fresh weights of roots and shoots, the contents of proline, soluble and insoluble sugars, ascorbic acid, and the activities of ascorbate peroxidase and polyphenol oxidase were evaluated. Seeds of chickpea, Cicer arietinum L. ILC482 variety were surface sterilized in sodium hypochlorite solution 1%, rinsed with sterilized water and germinated on moist filter papers in the dark for four days. Seedlings were transferred to plastic pots containing half-strength Hoagland solution, and were placed at 14 h photoperiod. Plants aged 14 days were randomly subdivided into two groups. One group received half-strength Hoagland solution (control) and another group was subjected half-strength Hoagland solution containing NO (0.1mM) for two days. Then, plants divided into three groups and, for 3 days, were subjected to 25/25, 25/15 and 25/5 ºC (day/night) regimes. After two days, shoots and roots were weighted for recording fresh weights. For assay of proline content, aliquots of fresh tissues were homogenized in 3% sulphosalicylic and centrifuged. Free proline contents were quantified using ninhydrin reagent and expressed as μmol/g FW. The total soluble sugars were determined by anthrone reaction at 625 nm in an 80% hot ethanol extract and expressed as mg/g FW. Later insoluble sugars were extracted from residues with HCl and determined by Anthrone reaction. In assay of ascorbate contents, 6% trichloroacetic acid extracted leaf tissues were mixed with 2,2-dipiridil. Then, for reduction of Fe3+ to Fe2+ by ascorbic acid mixture was incubated at 42 ºC and the absorbance values were recorded at 525 nm. Data expressed as μmol/g FW. For Enzyme assays, aliquots of fresh leaves were ground in cold extraction 100 mM phosphate buffers (pH 7.5) at 0-4 °C. After centrifugation of homogenates, enzymes assays were performed in the supernatant at 25 °C. Ascorbate peroxidase activity was measured by following the oxidation of ascorbate at290 nm. The Activity of Polyphenol oxidase in presence of pyrogallol was determined by measuring the increase in absorbance at 420 nm. The experiment was arranged in a completely randomized design with four replicates. The data were statistically analyzed by using Duncan's multiple range test to separate the means at p ≤ 0.05.


Results and Discussion
The fresh weights of roots and shoots, the contents of proline, soluble and insoluble sugars, ascorbic acid, and the activities of ascorbate peroxidase and polyphenol oxidase were evaluated. The results showed that nitric oxide pretreatment had a significant effect on the fresh weight of shoots, proline and soluble sugars contents, and polyphenol oxidase activity. Changes of night temperature were effective on all of the examined factors except shoots fresh weights. Interaction between nitric oxide and the temperature had a significant effect on fresh weights of roots and shoots, proline and soluble sugars contents, and ascorbate peroxidase activity. Data showed that maximum growth of plants occurred at 15 °C night temperature in the presence of NO. Proline contents of leaves were significantly decreased by falling night temperatures. NO treatments led to more reduction in proline under low night temperatures. In higher plants, proline is normally accumulated in response to stress factors. Night temperature reduction and NO treatment, probably, can act as signal for decreasing proline biosynthetic enzymes activities or for increasing proline degradative enzymes activities. Both soluble and insoluble sugars were increased significantly by falling night temperatures, and NO treatments had a positive effect. We can assume that falling of night temperatures can affect tissues metabolism, for preventing of damages, by accumulation of sugars. Falling of night temperatures increased ascorbate contents of leaves. Generally, ascorbate has antioxidative properties and high levels of foliar ascorbate can offer tolerance to plants under unfavorable conditions. Ascorbate peroxidase plays an important role in the metabolism of H2O2 in higher plants. In this study, falling of night temperatures led to decrease in the enzyme activity. However, enzyme activity increased significantly with the NO treatment at 5 °C. Ascorbate peroxidase activity is directly involved in the protection of plant cells against unfavorable environmental conditions.

Conclusion
In conclusion, falling night temperatures can significantly affect some biochemical markers of plants. The changes in roots and shoots are not showing the same patterns. Under low night temperatures, NO treatment can induce non enzymatic (ascorbate) and enzymatic (ascorbate peroxidase) defense systems for overcoming the deleterious effects of low temperature.

کلیدواژه‌ها [English]

  • Low temperature
  • Metabolism
  • Nitric oxide
  • Pea plant
1. Aghaee, A., Moradi, F., Zare-Maivan, H., Zarinkamar,F., Irandoost, H. P., and Sharifi, P. 2011. Physiological responses of two rice (Oryza sativa L.) genotypes to chilling stress at seedling stage. African Journal of Biotechnology 10(39): 7617-7621.
2. Akhar, F. K., Bagheri, A., Moshtaghi, N., and Nezami, A. 2011. The effect of gamma radiation on freezing tolerance of Chickpea (Cicer arietinum L.) at in vitro culture. Journal of Biological and Environmental Sciences 5: 63-70.
3. Azymi, S., Sofalian, O., Jahanbakhsh, G.S., and Khomari, S. 2012. Effect of chilling stress on soluble protein, sugar and proline accumulation in cotton (Gossypium hirsutum L.) genotypes. International Journal of Agriculture and Crop Sciences 4(12): 825-830.
4. Bates, L.S., Waldren, R.P., and Teare, I.D. 1973. Rapid determination of proline for water stress studies. Plant Soil 39: 205-207.
5. Boo, H.O., Heo, B.G., Gorinstein, S., and Chon, S.U. 2011.Positive effects of temperature and growth conditions on enzymatic and antioxidant status in lettuce plants. Plant Science 181: 479-484.
6. Chaparzadeh, N., D'Amico, M.L., Khavari-Nejad, R.A., Izzo, R., and Navari-Izzo F. 2004. Antioxidative responses of Calendula officinalis L. under salinity conditions. Plant Physiology and Biochemistry 42: 695-701.
7. Cho, S.C., Chao, Y.Y., Hong, C.Y., and Kao, C.H. 2012. The role of hydrogen peroxide in cadmium- inhibited root growth of rice seedling. Plant Growth Regulation 66: 27-35.
8. Chohan, A., Parmar, U., and Raina, S. K. 2012. Effect of sodium nitroprusside on morphological characters under chilling stress in chickpea (Cicer arietinum L.). Journal of Environmental Biology 33: 695-698.
9. Chon, S.U., Boo, H.O., Heo ,B.G., and Gorinstein, S. 2012. Anthocyanin content and the activities of polyphenol oxidase, peroxidase and phenylalanine ammonia-lyase in lettuce cultivars. International Journal of Food Sciences and Nutrition 63(1): 45-48.
10. Dabrowska, G., Kata, A., Goc, A., Szechynska-Hebda, M., and Skrzypek, E. 2007. Characteristics of the plant ascorbate peroxidase family. Acta Biologica Cracoviensia Series Botanica 49(1): 7-17.
11. Ganjewala, D., Boba, S., and Raghavendra, A.S. 2008. Sodium nitroprusside affects the level of anthocyanin and flavonol glycosides in pea (Pisum sativum L. cv. Arkel) leaves. Acta Biologica Szegediensis 52(2): 301-305.
12. Gill, P.K., Sharma, A.D., Singh, P., and Bhullar, S.S. 2001. Effect of various abiotic stresses on the growth soluble sugar and water relations of sorghum seedlings grown in light and darkness. Bulgarian Journal of Plant Physiology 27: 72-84.
13. Hare, P.D., Cress, W.A., and Staden, V. 1999. Proline synthesis and degradation: a model system for elucidating stress-related signal transduction. Journal of Experimental Botany 50: 413-434.
14. 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.
15. Hayat, S., Yadav, S., Wani, A.S., Irfan, M., Alyemini, M.N., and Ahmad, A. 2012. Impact of sodium nitroprusside on nitrate reductase, proline content, and antioxidant system in tomato under salinity stress. Horticulture, Environment and Biotechnology 53(5): 362-367.
16. Kovacik, J., Grz, J., Klejdus, B., Stork, F., Marchiosi, R., and Ferrarese-Filho, O. 2010. Lignification and related parameters in copper-exposed Matricaria chamomilla roots: role of H2O2 and NO in this process. Plant Science 179: 383-389.
17. Liu, X., Wang, L., Liu, L., Guo, Y., and Ren, H. 2011. Alleviating effect of exogenous nitric oxide in cucumber seedling against chilling stress. African Journal of Biotechnology 10: 4380-4386.
18. Mayer, A.M. 2006. Polyphenol oxidases in plants and fungi: Going places? Phytochemistry 67: 2318-2331.
19. Murgia, I., Tarantino, D., Vannini, C., Bracale, M., Carravieri, S., and Soave C. 2004. Arabidopsis thaliana plants overexpressing thylakoidalascorbate peroxidase show increased resistance to Paraquat induced photooxidative stress and to nitric oxide-induced cell death. The Plant Journal 38: 940-953.
20. Namvar, A., Sharif, R. S., and Khandan, T. 2011. Growth analysis and yield of chickpea (Cicer arietinum L.) in relation to organic and inorganic nitrogen fertilization. Ekologija 57: 97-108.
21. Palmieri, M.C., Sell, S, Huang, X., Scherf, M., Werner, T., Durner, J., and Lindermayer, C. 2008. Nitric oxide-responsive genes and promoters in Arabidopsis thaliana: a bioinformatics approach. Journal of Experimental Botany 59:177-186.
22. Rivero, R.M., Ruiz, J.M., Garcıa, P.C., Lopez-Lefebre, L.R., Sanchez, E., and Romero, L. 2001. Resistance to cold and heat stress: accumulation of phenolic compounds in tomato and watermelon plants. Plant Science 160: 315-321.
23. Rosa, M., Prado, C., Podazza, G., Interdonato, R., Gonzalez, J.A., Hilal, M., and Prado, F.E. 2009. Soluble sugars: metabolism, sensing and abiotic stress: a complex network in the life of plants. Plant Signaling and Behavior 4: 388-393.
24. Ruelland, E., and Zachowski, A. 2010. How plants sense temperature. Environmental and Experimental Botany 69: 225-232.
25. Shah, F., Huang, J., Cui, K., Nie, L., Shah, T., Wu, W., Wang, K., Khan, Z.H., Zhu, L., and Chen, C. 2011. Physiological and biochemical changes in rice associated with high night temperature stress and their amelioration by exogenous application of ascorbic acid (vitamin C). Australian Journal Crop Science 5(13): 1810-1816.
26. Siddiqui, M.H., Al-Whaibi, M.H., and Basalah, M.O. 2010. Role of nitric oxide in tolerance of plants to abiotic stress. Protoplasma 248(3): 447-455.
27. Simaei, M., Khavari-Nejad, R.A., Saadatmand, S., Bernard, F., and Fahimi, H. 2011. Effects of salicylic acid and nitric oxide on antioxidant capacity and proline accumulation in Glycine max L. treated with NaCl salinity. African Journal of Agricultural Research 6: 3775-3782.
28. Somogyi, M. 1952. Notes on sugar determination. Journal of Biological Chemistry 195: 19-23.
29. Strand, A., Hurry, V., Henkes, S., Huner, N., Gustafsson, P., Gardestrom, P., and Stitt, M. 1999. Acclimation of Arabidopsis leaves developing at low temperatures. Increasing cytoplasmic volume accompanies increased activities of enzymes in the Calvin cycle and in the sucrose biosynthesis pathway. Plant Physiology 119: 1387-1397.
30. Theocharis, A., Clement, C., and Barka, E.A. 2012. Physiological and molecular changes in plants grown at low temperatures. Planta 235(6): 1091-1105.
31. Wang, H., Zhang, S., Zhang, W., Wei, C., and Wang, P. 2010.Effects of nitric oxide on the growth and antioxidant response of submerged plants Hydrilla verticillata (Lf) Royle. African Journal of Biotechnology 9: 7470-7476.
32. Wang, Y., Wisniewski, M., Meilan, R., Cui, M., Webb, R., and Fuchigami, L. 2005. Overexpression of cytosolic ascorbate peroxidase in tomato confers tolerance to chilling and salt stress. Journal of the American Society for Horticultural Science 130(2): 167-173.
33. Windt, C.W., and Hasselt, P.R. 1999.Development of frost tolerance in winter wheat as modulated by differential root and shoot temperature. Plant Biology 1: 573-580.
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