الگوی بیان ژن های کدکننده آنزیم‌های کربنیک آنهیدراز، پراکسیداز و گلوتاتیون اس- ترانسفراز در گندم نان تحت شرایط کمبود روی (Zn)

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه ارومیه، ارومیه، ایران

2 استاد، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه ارومیه، ارومیه، ایران

3 استادیار، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه ارومیه، ارومیه، ایران

4 دانشجوی دکتری، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه ارومیه، ارومیه، ایران

چکیده

از استراتژی­های گیاهان در مقابله با تنش­ها، به­ویژه تنش‏های مربوط به کمبود ریزمغذی‏ها، استفاده از سیستم دفاعی آنتی­اکسیدان­های آنزیمی است. به­منظور بررسی اثر تنش کمبود روی بر بیان ژن­های کدکننده آنزیم­های کربنیک آنهیدراز (CAN)، پراکسیداز (PRX) و گلوتاتیون اس- ترانسفراز (GST) در ارقام روی- کارا (هامون) و روی- ناکارا (هیرمند) گندم نان، آزمایشی به­صورت فاکتوریل در قالب طرح کاملاً تصادفی با سه تکرار در گلخانه اجرا شد. ارقام در شرایط کمبود روی (صفر) و کفایت آن (5 میلی­گرم روی در کیلوگرم خاک) کشت و بیان نسبی این ژن‏ها در ریشه و برگ در دو مرحله رشدی شامل یک ماه بعد از جوانه­زنی (رویشی) و 30 درصد سنبله­دهی (زایشی) با روش Real time PCR اندازه­گیری شد. نتایج نشان داد که بیش­ترین میزان افزایش بیان ژن­ کربنیک آنهیدراز (211 برابر شاهد) در مرحله زایشی در ریشه رقم روی- کارای هامون مشاهده شد. این ژن در برگ نیز در مرحله زایشی بیان نسبی بالایی در هر دو رقم نشان داد. بیش­ترین میزان افزایش بیان ژن­های پراکسیداز (187 برابر شاهد) و گلوتاتیون اس- ترنسفراز (230 برابر شاهد) در برگ رقم روی- ناکارا (هیرمند) در مرحله زایشی مشاهده شد. به­طور کلی بیان هر سه ژن در ریشه رقم روی- کارا (هامون) در مرحله زایشی بیش­تر از رقم روی- ناکارا (هیرمند) بود. البته بیان ژن کربنیک آنهیدراز در برگ رقم روی- کارا نیز به­طور معنی­داری بیش­تر از رقم روی- ناکارا بود. نتایج این آزمایش نشان داد که در مرحله زایشی، ژنوتیپ­های روی- کارای گندم نان با افزایش بیش­تر بیان این ژن­ها در ریشه، به­طور مؤثرتری می­توانند نسبت به ژنوتیپ­های روی- ناکارا در برابر آسیب­های حاصل از تنش تحت شرایط کمبود روی مقابله کنند.

کلیدواژه‌ها


عنوان مقاله [English]

Expression pattern of genes encoding carbonic anhydrase, peroxidase and glutathione S-transferase enzymes in bread wheat under zinc deficiency conditions

نویسندگان [English]

  • Sahar Salehi 1
  • Babak Abdollahi Mandoulakani 2
  • Hadi Alipour 3
  • Kamran Moradi Gangachin 4
1 M. Sc. Student, Dept. of Plant Production and Genetics, Faculty of Agriculture, Urmia University, Urmia, Iran
2 Prof, Dept. of Plant Production and Genetics, Faculty of Agriculture, Urmia University, Urmia, Iran
3 Assist. Pro., Dept. of Plant Production and Genetics, Faculty of Agriculture, Urmia University, Urmia, Iran
4 Ph.D. Student, Dept. of Plant Production and Genetics, Faculty of Agriculture, Urmia University, Urmia, Iran
چکیده [English]

One of the plant strategies against stresses, especially micronutrient deficiencies, is the use of enzymatic antioxidant defense system. To investigate the effect of Zn deficiency on the genes expression of carbonic anhydrase (CAN), peroxidase (PRX) and glutathione S-transferase (GST) enzymes in Zn-efficient (Hamun) and -inefficient (Hirmand) bread wheat cultivars, a factorial experiment was carried out in completely randomized design (CRD) with three replications in greenhouse. The cultivars were grown in Zn deficient (zero) and adequacy (5 mg Zinc per kg soil) conditions and relative expression of the studied genes was measured using real time PCR technique in root and leaf of the cultivars at two growth stages, one month after germination (vegetative stage) and 30 % of flowering (reproductive stage). The results revealed the highest increase in CAN  (211 fold) expression in the root of Zn-efficient (Hamoon) cultivar in the reproductive stage. A realtively high expression of this gene was observed in the leaf of both cultivars in the reproductive stage. The highest increase in PRX (187 fold) and GST (230 fold)  expression was observed in the leaf of Zn-inefficient (Hirmand) cultivar at the reproductive stage. Generally, the expression of all three studied genes in the root of Zn-efficient (Hamoon) cultivar in the reproductive stage was higher than those of Zn-inefficient (Hirmand) cultivar. Also, the CAN expression in the leaf of Zn-efficient cultivar was significantly higher than that of Zn-inefficient cultivar. In conclusion, the results of the current study show that in the reproductive stage, Zn-efficient cultivars can effectively face against stress-induced damages through increasing the expression of the above-mentioned genes in the roots under Zn deficiency conditions.

کلیدواژه‌ها [English]

  • Antioxidant enzymes
  • Real time PCR
  • Zn-efficient cultivars
  • Zn-inefficient cultivar
Aizawa, K. and Miyachi, S. 1986. Carbonic anhydrase and CO2 concentrating mechanisms in microalgae and cyano-bacteria. Federation of European Microbiological Societies 39: 215-233.##Auld, D. S. 2001. Zinc coordination sphere in biochemical zinc sites. Biometals 14: 271-313.‏##Ayad, H. S., Reda, F. and Abdalla, M. S. A. 2010. Effect of putrescine and zinc on vegetative growth, photosynthetic pigments, lipid peroxidation and essential oil content of geranium (Pelargonium graveolens L.). World Journal of Agricultural Sciences 6 (5): 601-608.##Bar-Akiva, A. and Lavon, R. U. T. H. 1969. Carbonic anhydrase activity as an indicator of zinc deficiency in citrus leaves. Journal of Horticultural Science 44 (4): 359-362.##Baghban-Tabiat, S. and Rasouli-Sadaghiani, M. 2012. Investigation of Zn utilization and acquisition efficiency in different wheat genotypes at greenhouse conditions. Journal of Science and Technology of Greenhouse Culture 3 (2): 17-32. (In Persian with English Abstract).##Bandyopadhyay, T., Mehra, P., Hairat, S. and Giri, J. 2017. Morpho-physiological and transcriptome profiling reveal novel zinc deficiency-responsive genes in rice. Functional and Integrative Genomics 17 (5): 565-581.##Boyer, T. D. 1989. The glutathione S-transferases: An update. Hepatology 9 (3): 486-496.##Broadley, M. R., White, P. J., Hammond, J. P., Zelko, I. and Lux, A. 2007. Zinc in plants. New Phytology 173: 677-702.##Cakmak, I. 2000. Possible roles of zinc in protecting plant cells from damage by reactive oxygen species. The New Phytologist 146 (2): 185-205.##Cakmak, I. and Marschner, H. 1988. Enhanced superoxide radical production in roots of zinc deficient plants. Journal of Experimental Botany 39 (10): 1449-1460.##Campa, A. 1991. Biological roles of plant peroxidases: known and potential function. In: Everse, J. and Grisham, M. B. (Eds.). Peroxidases in chemistry and biology. Vol. 2. CRC Press. pp: 25-50.‏##Chen, W. R., He, Z. L., Yang, X. E. and Feng, Y. 2009. Zinc efficiency is correlated with root morphology, ultrastructure, and antioxidative enzymes in rice. Journal of Plant Nutrition 32(2): 287-305.##Cole, C. R., Grant, F. K., Swaby-Ellis, E. D., Smith, J. L., Jacques, A., Northrop-Clewes, C. A. and Ziegler, T. R. 2010. Zinc and iron deficiency and their interrelations in low-income African American and Hispanic children in Atlanta. Journal of Clinical Nutrition 91(4): 1027-1034.##Dunford, H. B. 1991. Horseradish peroxidase: Structure and kinetic properties. In: Everse, J. and Grisham, M. B. (Eds.). Peroxidases in chemistry and biology. Vol. 2. CRC Press. pp: 1-23.‏‏##Erenstein, O., Chamberlin, J. and Sonder, K. 2021. Estimating the global number and distribution of maize and wheat farms. Global Food Security 30: 100558.##Graham, R. D. and Rengel, Z. 1993. Genotypic variation in zinc uptake and utilization by plants. In: Robson, A. D. (Ed.). Zinc in soils and plants. Springer, Dordrecht. pp: 107-118.##Graham, R. D. and Welch, R. M. 1996. Breeding for staple food crops with high micronutrient density. Project paper. International Food Policy Research Institute. 79 p.##Graham, R. D., Ascher, J. S. and Hynes, S. C. 1992. Selecting zinc-efficient cereal genotypes for soils of low zinc status. Plant and Soil 146 (1-2): 241-250.##Gupta, B., Pathak, G. C. and Pandey, N. 2011. Induction of oxidative stress and antioxidant responses in Vigna mungo by zinc stress. Russian Journal of Plant Physiology 58 (1): 85-91.##Hacisalihoglu, G., Hart, J. J., Wang, Y. H., Cakmak, I. and Kochian, L. V. 2003. Zinc efficiency is correlated with enhanced expression and activity of zinc-requiring enzymes in wheat. Plant Physiology 131 (2): 595-602.##Helaly, A. A. E. and Ibrahim, F. R. 2019. Influence of iron, zinc and tyrosine acid on growth, yield components and chemical constituents of Hibiscus sabdariffa L. plant. Chemical Analysis 44 (34-10): 21-30.##Kampranis, S. C., Damianova, R., Atallah, M., Toby, G., Kondi, G., Tsichlis, P. N. and Makris, A.M. 2000. A novel plant glutathione S-transferase/peroxidase suppresses bax lethality in yeast. Journal of Biological Chemistry 275: 29207-29216.##Karpinski, S., Gabrys, H., Mateo, A., Karpinska, B. and Mullineaux, P. M. 2003. Light perception in plant diseasedefense signalling. Current Opinion in Plant Biology 6: 390-396.##Li, Y., Xie, S., Chen, Z. L., Xu, S. N. and Zhang, L. H. 2013. Impacts of zinc, benzo [a] pyrene, and their combination on the growth and antioxidant enzymes activities of wheat (Triticum aestivum L.) seedlings. Chinese Journal of Ecology 32 (2): 358-362.##Mahmoudi-Malhamlu, F. and Abdollahi-Mandoulakani, B. 2019. Enhanced expression of superoxide dismutase, phenylalanine ammonia-lyase and bZIP33 transcription factor encoding genes under Zn deficiency conditions in bread wheat (Triticum aestivum L.). Cereal Research
9 (1): 17-26. (In Persian with English Abstract).##Marschner, H. 2011. Mineral nutrition of higher plants. Academic Press, Cambridge, Massachusetts, USA.  89 p.##McGonigle, B., Keeler, S. J., Lau, S. M. C., Koeppe, M. K. and O’Keefe, D. P. 2000. A genomics approach to the comprehensive analysis of glutathione S-transferase gene family in soybean and maize. Plant Physiology 124: 1105-1120.##Mittler, R., Vanderauwera, S., Gollery, M. and Van-Breusegem, F. 2004. Reactive oxygen gene network of plants. Trends in Plant Science 9 (10): 490-498.##Moradi, K. and Abdollahi Mandoulakani, B. 2020. Expression pattern of HMA1, HMA2 and HMA9 genes under Zn deficiency conditions in bread wheat cultivars with different Zn uptake efficiency. Cereal Research 9 (4): 347-357. (In Persian with English Abstract).##Nakabayashi, R. and Saito, K. 2015. Integrated metabolomics for abiotic stress responses in plants. Current Opinion in Plant Biology 24: 10-16.##Niazkhani, S. M., Abdollahi-Mandoulakani, B., Jafari, M. and Rasouli-Sadaghiani, M. 2019. Effect of soil zinc deficiency on antioxidant enzymes activity and some biochemical parameters in bread wheat. Crop Physiology 11 (41): 27-5. (In Persian with English Abstract).##Pandey, N., Gupta, B. and Pathak, G. C. 2012. Antioxidant responses of pea genotypes to zinc deficiency. Russian Journal of Plant Physiology 59 (2): 198-205.##Parkin, G. 2004. Synthetic analogues relevant to the structure and function of zinc enzymes. Chemical Reviews 104 (2): 699-767.##Pfaffl, M. W. 2001. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Research 29 (9): 45-45.##Rahimi Jarihani, L. and Abdollahi Mandoulakani, B.  2021. Expression pattern of catalase, ascorbate peroxidase and polyphenol oxidase encoding genes under soil Zn deficiency in bread wheat. Cellular and Molecular Researches (Iranian Journal of Biology) 34 (1): 105-116. (In Persian with English Abstract).##Rengel, Z. 1995. Carbonic anhydrase activity in leaves of wheat genotypes differing in Zn efficiency. Journal of Plant Physiology 147 (2): 251-256.##Roxas, V. P., Smith, R. K., Allen, E. R. and Allen, R. D. 1997. Over expression of glutathione S-transferase / glutathione peroxidase enhances the growth of transgenic tobacco seedlings during stress. Nature Biotechnology 15: 988-991.##Sasaki, H., Hirose, T., Watanabe, Y. and Ohsugi, R. 1998. Carbonic anhydrase activity and CO2-transfer resistance in Zn-deficient rice leaves. Plant Physiology 118 (3): 929-934.##Sawaya, M. R., Cannon, G. C., Heinhorst, S., Tanaka, S., Williams, E. B., Yeates, T. O. and Kerfeld, C. A. 2006. The structure of β-carbonic anhydrase from the carboxysomal shell reveals a distinct subclass with one active site for the price of two. Journal of Biological Chemistry 281 (11): 7546-7555.##Seddigh, M., Khoshgoftarmanesh, A. H. and Ghasemi, S. 2013. The effectiveness of synthesized zinc-amino chelates in supplying zinc for wheat. Journal of Crop Production and Processing
3 (9): 177-187.‏##Selote, D. S. and Khanna-Chopra, R. 2010. Antioxidant response of wheat roots to drought acclimation. Protoplasma 245: 153-163.##Sies, H. and Stahl, W. 1995. Vitamins E and C, beta-carotene, and other carotenoids as antioxidants. The American Journal of Clinical Nutrition 62 (6): 1315S-1321S.‏##Subramanian, K. S., Tenshia, J. V., Jayalakshmi, K. and Ramachandran, V. 2011. Antioxidant enzyme activities in arbuscular mycorrhizal (Glomus intraradices) fungus inoculated and non-inoculated maize plants under zinc deficiency. Indian Journal of Microbiology 51 (1): 37-43.##Tenea, G. N., Bota, A. P., Raposo, F. C. and  Maquet, A. 2011. Reference genes for gene expression studies in wheat flag leaves grown under different farming conditions. BioMed Central Research Notes 4 (1): 373.‏##Welch, R. M. 2001. Impact of mineral nutrients in plants on human nutrition on a worldwide scale. Plant Nutrition-Food Security and Dordrecht, Netherlands. pp: 258-284.##