بررسی تغییرپذیری‌های الگوی بیان پروتئین در رقم‌های متحمل و حساس کلزا در شرایط تنش شوری

نوع مقاله : مقاله پژوهشی

نویسندگان

استادیار بخش کشاورزی، دانشگاه پیام نور، ایران

چکیده

به‌منظور بررسی سازوکار تحمل و حساسیت به تنش شوری در کلزا (Brassica napus L.)، رقم‌های SW5001 و Sarigol به ترتیب به‌عنوان رقم‌های متحمل و حساس به‌صورت آزمایش فاکتوریل در قالب طرح بلوک کامل تصادفی در چهار تکرار در گلخانه کشت شدند. نمونه‌های برگی، دو هفته پس از اعمال تنش شوری و در پایان مرحلۀ وردمانی (روزت) برداشت شدند. پروتئین از بافت برگی استخراج و الکتروفورز دوبعدی برای بررسی بیان پروتئین­ها در گیاهان شاهد و در شرایط تیمار شوری انجام شد. رنگ‌آمیزی ژل‌ها با آبی کوماسی و تجزیۀ لکه‌های پروتئینی با نرم‌افزار PDQuest انجام و لکه­های پروتئینی با استفاده از طیف‌سنجی (اسپکترومتری) جرمی و به روش MALDI TOF/TOF MS شناسایی شدند. شمار هفده لکۀ پروتئینی با اختلاف بیان معنی­دار بین گیاهان شاهد و تیمار تنش شوری، تشخیص داده شدند که از این شمار ده لکۀ پروتئینی بین دو رقم مشترک و شمار چهار و سه لکۀ پروتئینی به ترتیب به رقم متحمل و حساس اختصاص داشتند. پروتئین‌های مشترک شناسایی‌شده در گروه­های عملکردی شامل پروتئین‌های دخیل در واکنش نوری نورساخت (فتوسنتز)، چرخۀ کالوین، حذف پاداکسنده (آنتی­اکسیدانت)، انتقال پیام و پایداری ساختار پروتئین­ها طبقه‌بندی شدند. در مجموع در رقم متحمل SW5001 عامل‌های پایداری ساختار، حفظ کارایی سوخت‌وساز (متابولیسم) کربن و دفاع در برابر تنش اکسایشی در شرایط تنش به نحو مؤثری موجب ایجاد مقاومت شدند. در ضمن بیشترین نقطۀ قوت رقم متحمل با اتکا به پروتئین­های منحصربه‌فرد در قسمت واکنش نوری نورساخت بود و رقم حساس بیشترین آسیب را در چرخۀ کالوین متحمل شد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Assessment of changes in protein expression pattern in tolerant and sensitive cultivars of rapeseed under salt stress

نویسندگان [English]

  • Marouf Khalili
  • Mohammad Reza Naghavi
Assistant Professor, Department of Agriculture, Payamme Noor University, Iran
چکیده [English]

To investigate the mechanisms of tolerance and sensitivity to salinity stress in canola (Brassica napus L.), two spring canola cultivars such as SW5001 as tolerant cultivar and Sarigol as sensitive cultivar were evaluated at two levels of salinity in factorial based on randomized complete block design with four replications using a hydroponic culture system into green house. Sampling of leaves was performed in the rosette stage and two weeks after starting of salt stress. Proteins were extracted from leaf tissues and two-dimensional electrophoresis for the study of expression of proteins in both control and salt stressed plants was performed. Staining with commassie brilliant blue and quantitative analysis of protein spot by PDQuest software was performed and protein spots were identified by mass spectrometry using MALDI TOF/TOF MS method. 17 protein spots with significant expression differences between control and treatment plants to salinity, were identified that Of these, 10 protein spots was common between two cultivars and four and three proteins were assigned only to tolerant and sensitive cultivars respectively. Common proteins identified were classified in the functional groups of photo reaction of photosynthesis, Calvin cycle, proteins involved in remove of antioxidant, signal transduction and structural stability of proteins. In generally, in the tolerant cultivar of SW5001, sustainability factors of structure, maintaining of carbon metabolism efficiency and defense against to oxidative stress under stress conditions were effectively creates resistance. The highest strength point tolerant cultivar to rely on the unique proteins was in the light reaction of photosynthesis and the most of damage suffered in sensitive cultivar related to Calvin cycle.

کلیدواژه‌ها [English]

  • 2D electrophoresis
  • Canola
  • responsive proteins to stress
  • tolerance to salt stress

مراجع

  1. Ashraf, M. & Harris, P. J. C. (2004). Potential biochemical indicators of salinity tolerance in plants. Plant Science, 166, 3-16.
  2. Cakmak, I. (2005). The role of potassium in alleviating detrimental effects of abiotic stresses in plants. Journal of Plant Nutrition and Soil Science, 168, 521-530.
  3. Cui, S., Huang, F., Wang, J., Ma, X., Cheng, Y. & Liu, J. (2005). A proteomic analysis of cold stress responses in rice seedlings. Proteomics, 5(12), 3162-3172.
  4. Eberhard, S., Finazzi, G. & Wollman. F. A. (2008). The dynamics of photosynthesis. Annual Review of Genetics,42, 463-515.
  5. Gao, L., Yan, X., Li, X., Guo, G., Hu, Y., Ma, W. & Yan, Y. (2011). Proteome analysis of wheat leaf under salt stress by two-dimensional difference gel electrophoresis (2D-DIGE). Photochemistry, 72(10), 1180-1191.
  6. Guo, G., Ge, P., Ma, C., Li, X., Lv, D., Wang, S., Ma, W. & Yan, Y. (2012). Comparative proteomic analysis of salt response proteins in seedling roots of two wheat varieties. Journal of Proteomics, 75(6), 1867-1885.
  7. Hashimoto, M., Toorchi, M., Matsushita, K., Iwasaki Y. & Komatsu, S. (2009). Proteome analysis of rice root plasma membrane and detection of cold stress responsive proteins. Protein & Peptide Letters, 16, 685-697.
  8. Heide, H., Kalisz, H. M. & Follmann, H. (2004). The oxygen evolving enhancer protein 1 (OEE) of photosystem II in green algae exhibits thioredoxin activity. Journal of Plant Physiology, 161, 139-149.
  9. Herbert, B. (1999). Advances in protein solubilisation for two-dimensional electrophoresis. Electrophoresis, 20(4-5), 660-663.
  10. Holmstrom, K.O., Somersalo, S., Manda, A., Palva, T.E. & Welin, B. (2000). Improved tolerance to salinity and low temperature in transgenic tobacco producing glycine betaine. Journal of Experimental Botany, 51, 177-185.
  11. Hosseini Salekdeh, G. & Komatsu, S. (2007). Crop Proteomics: Aim at sustainable agriculture of tomorrow. Proteomics, 7, 2976-2996.
  12. Hosseini Salekdeh, Gh., Siopongco, J., Wade, L. J., Ghareyazie, B. & Bennett,  J. (2002). Proteomics analysis of rice leaves during drought stress and recovery. Proteomics, 2, 1131-1145.
  13. Ifuku, K., Ishihara, S., Shimamoto, R., Ido, K. & Sato, F. (2008). Structure, function, and evolution of the PsbP protein family in higher plants. Photosynthesis Research, 98, 427-437.
  14. Kausar, R., Arshad, M., Shahzad, A. & Komatsu, S. (2013). Proteomics analysis of sensitive and tolerant barley genotypes under drought stress. Amino Acids, 44, 345-359.
  15. Komatsu, S. & Tanaka, N. (2004). Rice proteome analysis: A step toward functional analysis of the rice genome. Proteomics, 4, 938-949.
  16. Kumara, S. G., Reddya, A. M. & Sudhakar, C. (2003). NaCl effects on proline metabolism in two high yielding genotypes of mulberry (Morus alba L.) with contrasting salt tolerance. Plant Science, 165, 1245-1251.
  17. Manna, A., Mimouni, H., Wasti, S., Gharbi, E., Aschi-Smiti, S., Faurobert, M. & Ben Ahmad, H. (2013) Comparative proteomic analysis of tomato (Solanum lycopersicum) leaves under salinity stress. Plant Omics Journal, 6, 268-277.
  18. Moller, A. L., Pedas, P., Andersen, B., Svensson, B., Schjoerring, J. K. & Finnie, C. (2011). Responses of barley root and shoot proteomes to longterm nitrogen deficiency, short-term nitrogen starvation and ammonium. Plant, Cell and Environment, 34(12), 2024-2037.
  19. Morant-Manceau, A., Pradier, E. & Tremblin, A. (2004). Osmotic adjustment, gas exchanges and chlorophyll fluorescence of a hexaploid triticale and its parental species salt stress. Journal of Plant Physiology, 169, 25-33.
  20. Naghavi, M. R. (2014). Evaluation of spring wheat cultivars under drought stress and proteome analysis for the most tolerant and sensitive ones. Ph.D. Thesis. Faculty of Agriculture, University of Tabriz. Iran. (in Farsi)
  21. Naghavi, M. R. (2010). Response and 2-Dimensional electrophoresis pattern of spring rapeseed genotypes under osmotic stress. M.Sc. Dissertation. Faculty of Agriculture, University of Tabriz, Iran. (in Farsi)
  22. Ozturk, M., Dzdemir, F., Eser, B., Adiyahsi, O. I. & Ilbi, H. (1995). Studies on the salt-hormone interactions in the germination and seedling growth of some vegetable species. In: M. Khan & I. A. Ingar (Eds.). Biology of salt tolerant plants. (pp.59-64.). University of Karach, Kharachi, Pakistan.
  23. Raymer, P. L. (2002). Canola: An emerging oilseed crop. In: J. Janick, & A. Whipkey, (Eds.). Trends in new crops and new uses. (pp. 44-56.). ASHS Press, Alexandria, VA.
  24. Reviron, M. P., Vartanian, N. M., Sallantin, J. C., Huet, J. C., Pernollet, J. & de Vienne, D. (1992). Characterization of a novel protein induced by progressive or rapid drought and salinity in Brassica napus leaves. Plant Physiology, 100(3), 1486-1493.
  25. Spreitzer, R. J. & Salvucci, M. E. (2002). Rubisco: structure, regulatory interactions, and possibilities for a better enzyme. Annual Review of Plant Biology, 53, 449-475.
  26. Sun, Y., Ahokas, R. A.,  Bhattacharya, S. K.,  Gerling, I. C., Carbone, L. D. & Weber, K. T. (2006). Oxidative stress in aldosteronism. Cardiovascular Research, 71, 300-309.
  27. Takahashi, S. & Murata, N. (2008). How do environmental stresses accelerate photo inhibition? Trends Plant Science, 13, 178-182.
  28. Tamoi, M., Nagaoka, M., Yabuta, Y. & Shigeoka, S. (2005). Carbon metabolism in the Calvin cycle. Plant Biotechnology, 22, 355-360.
  29. Thiellement, H., Zivy, M. & Plomion, C. (2002). Combining proteomic and genetic studies in plants. Chromatography B, 782, 137-149.
  30. Toorchi, M., Yukawa, K., Nouri, M. Z. & Komatsu, S. (2009). Proteomics approach for identifying osmotic-stress-related proteins in soybeans roots. Peptides, 30, 2108-2117.
  31. Twyman, R. M. (2004). Principles of proteomics (1st ed.). BIOS Scientific Publishers.
  32. Vaidyanathan, H., Sivakumar, P., Chakrabarty, R. & Thomas, G. (2003). Scavenging of reactive oxygen species in NaCl-stressed rice (Oryza sativa L.) - differential response in salt-tolerant and sensitive varieties. Plant Science, 165, 1411-1418.
  33. von Ballmoos, C. & Dimroth, P. (2007). Two distinct proton binding sites in the ATP synthase family. Biochemistry, 46, 11800-11809.
  34. Wan, X. Y. & Liu, J. Y. (2008). Comparative proteomics analysis reveals an intimate protein network provoked by hydrogen peroxide stress in rice seedling leaves. Molecular and Cellular Proteomics, 7, 1469-1488.
  35. Wang, W., Vinocur, B., Soseyov, O. & Altman, A. (2004). Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. Trends Plant Science, 9, 244-52.
  36. Xue, G. P., Mcintyre, C. L., Glassop, D. & Shorter, R. (2008). Use of expression analysis to dissect alterations in carbohydrate metabolism in wheat leaves during drought stress. Plant Molecular Biology, 67, 197-214.
  37. Yan, S., Su, Z., Tang, W. & Sun, W. (2005). Proteomic analysis of salt stress-responsive proteins in rice root. Proteomics, 5, 235-244.
  38. Ye, J., Wang, S., Zhang, F., Xie, D. & Yao, Y. (2013). Proteomic analysis of leaves of different wheat genotypes subjected to PEG6000 stress and rewatering. Plant Omics Journal, 6(4), 286-294.
  39. Yıldız, M., Akçalı, N. & Terzi, H. (2015). Proteomic and biochemical responses of canola (Brassica napus L.) exposed to salinity stress and exogenous lipoic acid. Journal of Plant Physiology, 179, 90-99.
  40. Zhu, M., Simons, B., Zhu, N., David, G., Oppenheimer, M. & Chen, S.  (2010). Analysis of abscisic acid responsive proteins in Brassica napus guard cells by multiplexed isobaric tagging. Journal of Proteomics, 73, 790-805.

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سابقه مقاله

  • تاریخ دریافت: 22 اردیبهشت 1395
  • تاریخ بازنگری: 22 دی 1395
  • تاریخ پذیرش: 25 دی 1395
  • تاریخ انتشار: 01 آذر 1396

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