بررسی اثر تنش شوری بر روی الگوی پروتئوم برگ در گندم Triticum boeoticum

نوع مقاله: علمی - پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد اصلاح نباتات، گروه بیوتکنولوژی، پژوهشکده علوم محیطی، پژوهشگاه علوم و تکنولوژی پیشرفته و علوم محیطی دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته، کرمان

2 استادیار گروه بیوتکنولوژی، پژوهشکده علوم محیطی، پژوهشگاه علوم و تکنولوژی پیشرفته و علوم محیطی دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته، کرمان

3 دانشیار اصلاح نباتات، گروه بیوتکنولوژی، دانشکده کشاورزی، دانشگاه شهید باهنر، کرمان

4 دانشیار، گروه بیوتکنولوژی، پژوهشکده علوم محیطی، پژوهشگاه علوم و تکنولوژی پیشرفته و علوم محیطی دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته، کرمان

5 استادیار بیوشیمی، گروه بیوتکنولوژی، پژوهشکده علوم محیطی، پژوهشگاه علوم و تکنولوژی پیشرفته و علوم محیطی، دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته، کرمان

6 کارشناس ارشد اصلاح نباتات، گروه اصلاح نباتات، دانشکده کشاورزی، دانشگاه تهران، کرج

چکیده

گیاهان وحشی خویشاوند گیاهان زراعی منابع مهمی برای یافتن ژن ­های اعطاءکننده تحمل به تنش شوری هستند. در این مطالعه ده توده مختلف گندم بوئتیکوم در گلخانه به­صورت آزمایش فاکتوریل در قالب طرح کاملاً تصادفی کشت گردیدند. در مرحله سه برگی تنش شوری در دو سطح صفر و 150 میلی­ مولار اعمال و بعد از 15 روز نمونه ­برداری از پهنک جوان ترین برگ توسعه­ یافته انجام گرفت. یون‌های سدیم و پتاسیم در هر نمونه اندازه ­گیری و سپس متحمل‌ترین جمعیت براساس غلظت یون سدیم و صفت K+/Na+  مشخص شد. نمونه حاصل از گندم متحمل برای انجام الکتروفورز دو­بعدی مورد استفاده قرار گرفت. پروتئین‌ها پس از استخراج ابتدا با استفاده از نوارهای IPG با pH 3-10 براساس نقطه ایزوالکتریک و سپس در بعد دوم با استفاده از SDS-PAGE براساس وزن مولکولی جداسازی گردیدند. نتایج نشان داد که دو توده C11 و C7 جمع­ آوری شده از مناطق سقز و کامیاران متحمل‌ترین و دو توده B2 و C1 جمع­آوری شده از مناطق هرسین و سنندج حساس­ترین بودند. براساس صفات فیزیولوژیک نمونه C11 برای انجام پروتئومیکس انتخاب گردید. نتایج نشان داد که تعداد 177 لکه تکرارپذیر در ژل‌ها شناسایی و مورد تجزیه آماری قرار گرفتند. از این تعداد 13 لکه تحت شرایط تنش شوری تغییر بیان نشان دادند که از بین آنها 8 لکه (61/5%) افزایش بیان و 5 لکه (38/5%) کاهش بیان نشان دادند. این بدان معنی است که C11 با تغییرات در بیان ژن‌های پاسخ­ دهنده سعی در مقابله با تنش شوری 150 میلی ­مولاری و تحمل آن را داشته است. 

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Investigation of Salinity Effect on Leaf Proteome Pattern of Triticum boeoticum

نویسندگان [English]

  • Najmeh Sadat Alavi 1
  • Mahmood Maleki 2
  • Shahram Pourseiedi 3
  • Mehdi Rahimi 2
  • Amin Baghizadeh 4
  • Ali Riahi Medvar 5
  • Abdol-Rahman Rasoulnia 6
1 M.Sc. Student of Plant Breeding, Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman
2 Assistant Professor, Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman
3 Associate Professor of Plant Breeding, Faculty of Agriculture, Shahid Bahonar University, Kerman
4 Associate Professor, Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman
5 Assistant professor of Biochemistry, Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman
6 M.Sc. of Plant Breeding, Department of Plant Breeding, Faculty of Agriculture,Tehran University, Karaj
چکیده [English]

Wild relatives of crop plants is an important resource for finding the genes conferring tolerance to salinity. In this study, ten different ecotypes of Triticum boeoticum were planted in greenhouse by using factorial experiment based on completely randomized design. Salt stress was applied at the three-leaf stage and at two levels: zero and 150 mM and were sampled after 15 days from newly full developed blade leaf. The concentration of sodium and potassium were measured in each sample and after that the tolerant ecotype was determined based on concentration of Na+ and K+/Na+ trait. Leaf sample of tolerant ecotype were used for two dimensional electrophoresis. Extracted proteins first isolated based on isoelectric point by using IPG strip with pH 3-10 and then isolated based on molecular weight in the second dimension. The result showed that C11 and C7 that collected from Saghez and Kamiaran respectively were most tolerant populations. Both of B2 and C1 that collected from Harsin and Sanandaj respectively were most sensitive populations. Based on physiological traits, sample C11 was selected for proteomics. According to the proteomics results, the number of repetitive 177 gel spots were identified and statistically analyzed. From total spots, 13 spots showed differential expression under salt stress, from these, the expression of 8 spots (61.5%) increased and 5 spots (38.5%) decreased under salt stress. This means that the C11 is trying to cope with salinity stress by changing the expression of their responsive genes. 

کلیدواژه‌ها [English]

  • Salt stress
  • Triticum boeoticum
  • proteomics

Appel, R. D., Palagi, P. M., Walther, D., Vargas, J. R., Sanchez, J. C., Ravier, F., Pasquali, C. and Hochstrasser, D. F. 1997. Melanie II–a third‐generation software package for analysis of two‐dimensional electrophoresis images: I. Features and user interface. Electrophoresis, 18(15): 2724-2734.

Asch, F., Dingkuhn, M., Dörffling, K. and Miezan, K. 2000. Leaf K/Na ratio predicts salinity induced yield loss in irrigated rice. Euphytica, 113(2): 109-118.

Ashraf, M. and Haris, P. J. C. 2004. Potential biochemical indicatora of salinity tolerance in plant. Plant Science, 166(1): 3-16.

Askari, H., Edqvist, J., Hajheidari, M., Kafi, M. and Hosseini Salekdeh, G. 2006. Effects of salinity levels on proteome of Suaeda baegyptiaca leaves. Proteomics, 6: 2542-2554.

Bhandal, I. S. and Malik, C. P. 1988. Potassium estimation, uptake, and its role in the physiology and metabolism of flowering plants. International Review of Cytology, 110: 205-254.

Blum, H., Beier, H. and Gross, H. J. 1987. Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis, 8(2): 93-99.

Bohnert, H. J., Nelson, D. F. and Jenson, R. G. 1995. Adaptation to environmental stresses. Plant Cell, 7: 1099-1111.

Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1-2): 248-254.

Brini, F., Hanin, M., Mezghani, I., Berkowitz, G. A. and Masmoudi, K. 2007. Overexpression of wheat Na+/H+ antiporter TNHX1 and H+-pyrophosphatase TVP1 improve salt-and drought-stress tolerance in Arabidopsis thaliana plants. Journal of Experimental Botany, 58(2): 301-308.

Capriotti, A. L., Borrelli, G. M., Colapicchioni, V., Papa, R., Piovesana, S., Samperi, R., Stampachiacchiere, S. and Laganà, A. 2014. Proteomic study of a tolerant genotype of durum wheat under salt-stress conditions. Analytical and Bioanalytical Chemistry, 406: 1423-1435.

Caruso, G., Cavaliere, C., Guarino, C., Gubbiotti, R., Foglia, P. and Laganà, A. 2008. Identification of changes in Triticum durum L. leaf proteome in response to salt stress by two-dimensional electrophoresis and MALDI-TOF mass spectrometry. Analytical and Bioanalytical Chemistry, 391(1): 381-390.

Caruso, T., Chan, Y., Lau, M. C. Y., McKay, C. P. and Pointing, S. B. 2011. Stochastic and deterministic processes interact in the assembly of desert microbial communities on a global scale. International Society for Microbial Ecology (ISME) Journal, 5: 1406-1413.

Chhipa, B. R. and Lal, P. 1995. Na/K ratios as the basis of salt tolerance in wheat. Crop and Pasture Science, 46: 533-539.

Churin, Y., Hess, W. R. and Borner, T. 1999. Cloning and characterization of three cDNAs encoding chloroplast RNA-binding proteins from barley (Hordeum vulgare L.): differential regulation of expression by light and plastid development. Current Genetics, 36: 173-181.

Damerval, C. D. V., Zivy, D. and Thiellement, H. 1986. Technical improvements in two dimensional electrophoresis increase the level of genetic variation detected in wheat seedling proteins. Electrophoresis, 7(1): 52-54.

Flowers, T. J. and Yeo, A. R. 1986. Ion relations of plants under drought and salinity. Functional Plant Biology, 13: 75-91.

Francois, L. E., Maas, E. V., Donovan, T. J. and Youngs, V. L. 1986. Effect of salinity on grain yield and quality, vegetative growth, and germination of semi-dwarf and durum wheat. Agronomy Journal, 78: 1053-1058.

Gao, L., Yan, X., Li, X., Guo, G., Hua, Y., Ma, W. and Yan, Y. 2011. Proteome analysis of wheat leaf under salt stress by two-dimensional difference gel electrophoresis (2D-DIGE). Phytochemistry, 72(10): 1180-1191.

Garcia, A., Senadhira, D., Flowers, T. J. and Yeo, A. R. 1995. The effects of selection for sodium tt-ansport and of selectior-r for agronomic char-actet-islies upon salt resistance in rice (Oryza sativa L.), Theoretical and Applied Genetics, 90: 1106-1 111.

Gorg, A., Boguth, G. H., Scheibe, A., Wildgruber, B. and Weiss, R. W. 2000. The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis, 21: 1037-1053.

Gorham, J., Hardy, C., Wyn Jones, R. G., Joppa, L. R. and Law, C. N. 1987. Chromosomal location of K+/Na+ discrimination character in the D genome of wheat. Theoretical and Applied Genetics, 74: 584-588.

Gygi, S. P. and Aebersold, R. 2000. Mass spectrometry and proteomics. Current Opinion in Chemical Biology, 4: 489-494.

Gygi, S. P., Rochon, Y., Franza, B. R. and Aebersold, R. 1999. Correlation between protein and mRNA abundance in yeast. Molecular and Cellular Biology, 19: 1720-1730.

James, R. A., Davenport, R. J. and Munns, R. 2006. Physiological characterization of two genes for Na+ exclusion in durum wheat, Nax1 and Nax2. Plant Physiology, 142(4): 1537-1547.

Maathuis, F. J. M. and Amtmann, A. 1999. K+ nutrition and Na+ toxicity: the basis of cellular K+/Na+ ratios. Annals of Botany, 84: 123-133.

Maleki, M., Naghavi, M. R., Alizadeh, H., Poustini K. and Abd Mishani, C. 2012. Effect of salinity on changes of protein profile in seedlings of wheat (Triticum aestivum L.) cv. Roshan. Iranian Journal of Crop Sciences, 13(4): 684-696. (In Persian).

Maleki, M., Naghavi, M. R., Alizadeh, H., Poostini, K. and Mishani, C. A. 2014. Comparison of protein changes in the leaves of two bread wheat cultivars with different sensitivity under salt stress. Annual Research & Review in Biology, 4(11): 1784-1797.

Marschner, H. 1995. The Mineral Nutrition of Higher Plants. Academic Press, London.

Munns, R. 1993. Physiological processes limiting plant growth in saline soils: Some dogmas and hypotheses. Plant, Cell & Environment,16: 15-24.

Munns, R. and James, R. A. 2003. Screening methods for salt tolerance: a case study with tetraploid wheat. Plant and Soil, 253: 201-218.

Noble, C. L. and Rogers, M. E. 1992. Arguments for the use of physiological criteria for improving the salt tolerance in crops. Plant Soil, 146: 99-107.

Rengasamy, P. 2002. Transient salinity and subsoil constraints to dryland farming in Australian sodic soils: an overview. Australian Journal of Experimental Agriculture, 42(3): 351-361.

Shabala, S. and Cuin, T. A. 2008. Potassium transport and plant salt tolerance. Physiologia Plantarum, 133(4): 651-669.

Tester, M. and Davenport, R. 2003. Na+ tolerance and Na+ transport in higher plants. Annals of Botany, 91(5): 503-527.

Waines, J. G. 1983. Genetic resources in diploid wheats: The case for diploid Commercial wheats. In Proceedings of the sixth International Wheat Genetics Symposium/edited by Sadao Sakamoto. Kyoto: Plant Germ-Plasm Institute, Faculty of Agriculture, Kyoto University, Pp. 115-122.

Wang, W., Vinocur, B. and Altman, A. 2003. Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta, 218 (1): 1-14.

Yeo, A. R., Flao, S. A., Welfare, K., Senanayake, N. and Flowers, T. J. 1999. Silicon reduces sodium uptake in rice (Oryza sativa L.) in saline conditions and this is accounted for by a reduction in the transpirational bypass flow. Plant, Cell and Environment, 22: 559-565.

Yıldız, M. 2007. Two-dimensional electrophoretic analysis of soluble leaf proteins of a salt-sensitive (Triticum aestivum) and a salt-tolerant (Triticumdurum) cultivar in response to NaCl stress. Journal of Integrative Plant Biology, 49: 975-981.

Zivy, M. and de Vienne, D. 2000. Proteomics: a link between genomics, genetics and physiology. Plant Molecular Biology, 44: 575-580.