با همکاری مشترک دانشگاه پیام نور و انجمن بیوتکنولوژی جمهوری اسلامی ایران

نوع مقاله : علمی پژوهشی

نویسندگان

1 استادیار بخش کشاورزی، دانشگاه پیام نور، تهران، ایران

2 دانشیار بخش کشاورزی، دانشگاه پیام نور، تهران، ایران

چکیده

به منظور بررسی مکانسیم تحمل به تنش شوری در ارقام گندم، آزمایشی فاکتوریل بر پایه طرح کاملاً تصادفی با چهار تکرار در محیط گلخانه انجام شد. عوامل این آزمایش شامل تنش شوری از نوع کلریدسدیم در چهار سطح صفر (شاهد)، 100، 200 و 300 میلی‌مول و دو رقم گندم نیک‌نژاد و پیشتاز بترتیب بعنوان نماینده‌ای از ارقام متحمل و حساس بودند. نمونه‌های برگی، دو هفته پس از اعمال تنش شوری تهیه شدند. سپس استخراج پروتئین از بافت برگی صورت گرفت و الکتروفورز دوبعدی در گیاهان شاهد و تحت تیمارهای تنش شوری انجام شد. مقایسه نتایج تجزیه پروتئوم در سطوح تنش نشان داد که تعداد 15 لکه پروتئینی تکرارپذیر با تغییر بیان متفاوت بین دو رقم متحمل و حساس مشترک بوده و تعداد پنج لکه پروتئینی منحصر به هر رقم متحمل و حساس تحت تنش تغییر بیان معنی‌دار داشتند. لکه‌های پروتئینی با استفاده از طیف‌سنجی جرمی شناسایی شدند و نتایج نشان داد که پروتئین‌های مشترک شناسایی شده بیشتر در گروه‌های عملکردی شامل دفاع آنتی‌اکسیدانی و چرخه کالوین طبقه‌بندی شدند. در حالی که سایر پروتئین‌ها در هر رقم بیشتر در فعالیت آنتی-اکسیدانی نقش داشتند. در مجموع نتایج نشان داد که بین دو رقم مورد مطالعه از لحاظ پاسخ مورفو-فیزیولوژیکی به تنش شوری تفاوت معنی‌دار وجود دارد و رقم متحمل نیک‌نژاد پاسخ پروتئینی مناسبتری تحت تنش نشان می‌دهد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Study morpho-physiological change and protein pattern of tolerant and susceptible wheat cultivars under salinity stress by using proteome analysis

نویسندگان [English]

  • Mohammad Reza Naghavi 1
  • Marouf Khalili 2

1 Assistant Professor, Department of Agriculture, Payame Noor University, Tehran, Iran.

2 Associate Professor, Department of Agriculture, Payame Noor University, Tehran, Iran

چکیده [English]

In order to investigate the mechanism of tolerance to salt stress in wheat, a factorial experiment basis of completely randomized design with four replications in greenhouse media were conducted. The factors of this experiment were included salinity of sodium chloride in four levels of zero (control), 100, 200 and 300 mM and two wheat cultivars namely Niknejhad and Pishtaz as a representative of tolerant and susceptible cultivars, respectively. Leaf samples were prepared two weeks after the starting of salt stress. Then, extraction of protein from leaf tissue was done and two-dimensional electrophoresis in control plants and under salt stress plants were carried out. The results of proteome analysis revealed that 15 replicated protein spots with different expression variations were common between two tolerant and susceptible cultivars, and five protein spots of unique to each tolerant and susceptible cultivar, had significantly expression variations under stress. Protein spots were detected by mass spectrometry and the results showed that more number from commonly detected proteins were classified in functional groups including antioxidant defense and Calvin cycle, while other proteins in each cultivar had more roles in antioxidant activity. Overall, the results showed that there was a significant difference between two cultivars in terms of morpho-physiological response to salt stress, and the Niknejhad cultivar showed a better protein response under stress.

کلیدواژه‌ها [English]

  • Proteome analysis
  • responsive proteins to stress
  • tolerance of salt stress
  • Wheat
Albertin W, Langella O, Joets J, Negroni L, Zivy M, Damerval C, Thiellement H (2009) Comparative proteomics of leaf, stem, and root tissues of synthetic Brassica napus. Proteomics, 9: 793-799.
Ali GM, Komatsu S (2006) Proteomic Analysis of Rice Leaf Sheath during Drought Stress. J. Proteome Res. 5(2): 396-403.
Araus JL, Ceccarelli S, Grando S (1997) Relationship between leaf structure and carbon isotope discrimination in field-grown barley. Plant Physiol. Biochem. 35: 533-541.
Arias D (2007) Calibration of LAI-2000 to Estimate Leaf Area Index and Assessment of its Relationship with stand productivity in six Native and Introduced tree Species in costarica. Forest Ecol. Manag. 247: 85-193.
Asada K (1992) Ascorbate peroxidase hydrogen peroxide scavenging enzyme in plants. Physiologica Plantarum, 85: 235-241.
Bajjii M, Lutts S, Kinet KM (2001) Water deficit effects on solute contribution to osmotic adjustment as a function of leaf ageing in three durum wheat (Triticum durum Desf) cultivars performing in arid conditions. Plant Sci. 60: 669-681.
Berardini TZ, Mundodi S, Reiser R, Huala E, Garcia-Hernandez M, Zhang P, Mueller LM, Yoon J, Doyle A, Lander G, Moseyko N, Yoo D, Xu I, Zoeckler B, Montoya M, Miller N, Weems D, Rhee SY (2004) Functional annotation of the Arabidopsis genome using controlled vocabularies. Plant Physiol. 135:745-755.
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 7(72): 248-254.
Cakmak I (2005) The role of potassium in alleviating detrimental effects of abiotic stresses in plants. J. Plant Nutr. Soil Sci. 168: 521-530.
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.
Dhindsa RS (1991) Drought stress, enzymes of glutathione metabolism, oxidation injury, and protein synthesis in Tortula ruralis. Plant Physiol. 95: 648-651.
Ding Y, Ma QH (2004) Characterization of a cytosolic malate dehydrogenase cDNA which encodes an isozyme toward oxaloacetate reduction in wheat. Biochimie, 86: 509-518.
Edwards R, Dixon DP, Walbot V (2000) Plant glutathione S-transferases: enzymes with multiple functions in sickness and in health. Trends Plant Sci. 5: 193-198.
Ford KL, Cassin A, Bacic A (2011) Quantitative proteomic analysis of wheat cultivars with differing drought stress tolerance. Plant Sci. 2(44): 1-11.
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). ‎Photochem. 72(10): 1180-1191.
Gelhaye E, Rouhier N, Navrot N, Jacquot JP (2005) The plant thioredoxin system, Cell Mol. Life Sci., 62:24-35.
Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol. Biochem. 48(12): 909-930.
Guan HP, Keeling PL (1998) Starch Biosynthesis: Understanding the functions and interactions of multiple isoenzymes of starch synthase and branching enzyme. Trends Glycosci Glyc. 10: 307-319.
Guicherd, P., Peltier, J.P., Gout, E., Bligny, R. and Marigo, G. (1997) Osmotic adjustment in Fraxinus excelsior L. malate and mannitol accumulation in leaves under drought conditions. Trees, 11: 155-161.
Han F, Chen H, Yan MF, Liu GS, Shen SH (2009) A comparative proteomic analysis of rice seedlings under varioushigh-temperature stresses. Biochimica et Biophysica Acta., 1794: 1625-1634.
Hasegawa PM, Bressan RA, Zhu JK, Bohnert HJ (2000) Plant cellular and molecular responses to high salinity. Annu. Rev. Plant Physiol. Plant Mol. Biol. 51: 463-499.
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 Pept. Lett. 16: 685-697.
Heide H, Kalisz HM, Follmann H (2004) The oxygen evolving enhancer protein 1 (OEE) of photosystem II in green algae exhibits thioredoxin activity. J. Plant Physiol. 161: 139-149.
Heldt HW (1997) Plant biochemistry and molecular biology. Oxford: Oxford University Press.
Herbert B (1999) Advances in protein solubilisation for two-dimensional electrophoresis. Electrophoresis, 20(4-5): 660-663.
Holmstrom KO, Somersalo S, Manda A, Palva TE, Welin B (2000) Improved tolerance to salinity and low temperature in transgenic tobacco producing glycine betaine. J. Exp. Bot. 51: 177-185.
Hosseini Salekdeh Gh, Siopongco J, Wade LJ, Ghareyazie B, Bennett J (2002) Proteomics analysis of rice leaves during drought stress and recovery. Proteomics, 2: 1131-1145.
Ifuku K, Ishihara S, Shimamoto R, Ido K, Sato F (2008) Structure, function, and evolution of the PsbP protein family in higher plants. Photosynth. Res. 98: 427-437.
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.
Kiddle G, Bennett R, Hick A, Wallsgrove R (1999) C-S lyase activities in leaves of crucifers and non-crucifers, and the characterization of three classes of C-S lyase activities from oilseed rape (Brassica napus L.). Plant Cell Environ. 22: 433-445.
Kim SH, Mizuno K, Fujimura T (2002) Regulated expression of ADPglucose pyrophosphorylase and chalcone synthase during root development in sweet potato. Plant Growth Regul. 38: 173-179
Komatsu S, Tanaka N (2004) Rice proteome analysis: A step toward functional analysis of the rice genome. Proteomics, 4: 938-949.
Kruger NJ (1997) Carbohydrate synthesis and degradation. . In: Dennis, D.T., Turpin, D.H Lefebvre, D.D. and Layzell, D.B. (Eds.), Plant Metabolism (2nd ed., pp. 83-104). Essex: Addison Wesley Longman.
Macdonald, F.D. and Buchanan, B.B. (1997) The reductive pentose phosphate pathway and its regulation. In: Dennis DT, Turpin DH, Lefebvre DD, Layzell DB (Eds.), Plant Metabolism (2nd ed., pp. 299-313). Essex: Addison Wesley Longman.
Martinez JP, Luttus S, Schanck A, Banjji M (2004) Is osmotic adjustment required for water stress resistance in the Mediterranean shrub Atriplexhalimus L. J. Plant Physiol. 16: 1041-1051.
Minarik P, Tomaskova N, Kollarova M, Antalik M (2002) Malate dehydrogenases-structure and function. General Physiol. Biophys. 21: 257-265.
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. J. Plant Physiol. 169: 25-33.
Musrati RA, Kollarova M, Mernik N, Mikulasova D (1998) Malate dehydrogenase: distribution, function and properties. Gen. Physiol. Biophys. 17: 193-210.
Naghavi MR (2014) Evaluation of spring wheat cultivars under drought stress and proteome analysis for the most tolerant and sensitive ones. PhD Thesis. Faculty of Agriculture. University of Tabriz. Iran (in Farsi)
Okcu G, Kaya MD, Atak M (2005) Effect of salt and drought stress on germination and seedling growth of pea (Pisum sativum). Turk. J. Agric. 29: 137-243.
Petrov VD, Van Breusegem F (2012) Hydrogen peroxide-a central hub for information flow in plant cells. AoB plants 2012. pls014.
Pillai MA, Lihuang Z, Akiyama T (2002) Molecular cloning, characterization, expression and chromosomal location of OsGAPDH, a submergence responsive gene in rice (Oryza sativa L.). Theor. Appl. Genet. 105: 34-42
Plucken H, Muller B, Grohmann D, Westhoff P, Eichacker LA (2002) The HCF136 proteinis essential for assembly of the photosystem II reaction center in Arabidopsis thaliana. FEBS Lett. 532: 85-90.
Sairam RK, Rao KV, Srivastava GC (2002) Differential response of wheat genotypes to long term salinity stress in relation to oxidative stress, antioxidant activity and osmolyte concentration. Plant Sci. 163: 1037-1046.
Samaj J, Thelen JJ (2007) Plant proteomics. In: Samaj J, Thelen JJ (Eds.). Plant proteomics. Institute of Plant Genetics and Biotechnology, Slovak Republic.
Spreitzer RJ, Salvucci ME (2002) Rubisco: structure, regulatory interactions, and possibilities for a better enzyme. Annu. Rev. Plant Biol. 53: 449-475.
Sun Y, Ahokas RA, Bhattacharya SK, Gerling IC, Carbone LD, Weber KT (2006) Oxidative stress in aldosteronism. Cardiovasc. Res. 71: 300-309.
Takahashi S, Murata N (2008) How do environmental stresses accelerate photo inhibition? Trends Plant Sci. 13: 178-182.
Tamoi M, Nagaoka M, Yabuta Y, Shigeoka S (2005) Carbon metabolism in the Calvin cycle. Plant Biotechnol. 22: 355-360.
Tamura T, Hara K, Yamaguchi Y, Koizumi N, Sano H (2003) Osmotic stress tolerance of transgenic tobacco expressing a gene encoding a membrane-located receptor-like protein from tobacco plants. Plant Physiol. 131: 454-462.
Thiellement H, Zivy M, Plomion C (2002) Combining proteomic and genetic studies in plants. Chromatography B, 782: 137-149.
Tetlow IJ, Davies EJ, Vardy KA, Bowsher CG, Burrell MM, Emes MJ (2003) Subcellular localization of ADPglucose pyrophosphorylase in developing wheat endosperm and analysis of the properties of a plastidial isoform. J. Exp. Bot., 54: 715-725.
Tewari RK, Hadacek F, Sassmann S, Lang I (2013) Iron deprivation-induced reactive oxygen species generation leads to non-autolytic PCD in Brassica napus leaves. Environ. Exp. Bot. 91: 74-83.
Twyman RM (2004) Principles of proteomics. BIOS Scientific Publishers.
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 Sci. 165: 1411-1418.
von Ballmoos C, Dimroth P (2007) Two distinct proton binding sites in the ATP synthase family. Biochem. 46: 11800-11809.
Wan XY, Liu JY (2008) Comparative proteomics analysis reveals an intimate protein network provoked by hydrogen peroxide stress in rice seedling leaves. Mol. Cell Proteomics, 7: 1469-1488.
Wang MC, Peng ZY, Li Cl, Li F, Liu C, Xia GM (2008) Proteomic analysis on a high salt tolerance introgression strain of Triticum aestivum/ Thinopyrum ponticum. Proteomics, 8: 1470-1489.
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 Sci. 9: 244-52.
Yadav SK, Singla-Pareek SL, Ray M, Reddy MK, Sopory SK (2005) Methylglyoxal levels in plants under salinity stress are dependent on glyoxalase I and glutathione. Biochem. Biophys. Res. Commun. 337: 61-67.
Yang YJ (2008) Proteome comparative analysis of Cryptochromes of Arabidopsis. Hunan University, Changsha.
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.
Yildiz M (2007) Two-Dimensional Electrophoretic Analysis of Soluble Leaf Proteins of a Salt-sensitive (Triticum aestivum) and a Salt-tolerant (T. durum) Cultivar in Response to NaCl Stress. J. Integr. Plant Biol. 49(7): 975-981.
Yıldız M, Akçalı N, Terzi H (2015) Proteomic and biochemical responses of canola (Brassica napus L.) exposed to salinity stress and exogenous lipoic acid. J. Plant Physiol. 179: 90-99.
Yong Z, Hao-Ru T, Ya L (2008) Variation in antioxidant enzyme activities of two strawbreey cultivars with short-term low temperature stress. J. Agric. Sci. 4: 456-462.
Zang X, Komatsu S (2007) A proteomic approach for identifying osmotic-stress-relatedproteins in rice. Phytochem. 68: 426-437.