Volume 4, Issue 6, November 2016, Page: 78-90
The mtlD Gene-overexpressed Transgenic Wheat Tolerates Salt Stress Through Accumulation of Mannitol and Sugars
Mohamed Ahmed Seif El-Yazal, Botany Department, Faculty of Agriculture, Fayoum University, Fayoum, Egypt
Hala Fawzi Eissa, Agricultural Genetic Engineering Research Institute (AGERI), Agriculture Research Center (ARC), Giza, Egypt
Safia Mahmoud Abd El-Mageed Ahmed, Botany Department, Faculty of Agriculture, Fayoum University, Fayoum, Egypt
Saad Mohamed Howladar, Biology Department, Faculty of Sciences, Albaha University, Albaha, Saudi Arabia
Safi-naz Sabet Zaki, Department of Water Relations and Field Irrigation, National Research Centre, Dokki, Cairo, Egypt
Mostafa Mohamed Rady, Botany Department, Faculty of Agriculture, Fayoum University, Fayoum, Egypt
Received: Sep. 17, 2016;       Accepted: Oct. 12, 2016;       Published: Oct. 31, 2016
DOI: 10.11648/j.plant.20160406.15      View  2125      Downloads  61
Abstract
The mtlD gene-contained transgenic wheat has established the role of mannitol and sugars accumulation in alleviating the abiotic stresses, including salinity. This study was conducted to determine whether the 85 mM NaCl-salinity could be tolerated by wheat (genotype 235/3) plants of which seeds were transformed with mtlD gene (from Escherichia coli). The effects of mtlD gene transformation into wheat seeds on growth traits, physio-biochemical attributes, and yield and its quality of transgenic wheat genotype were investigated compared to non-transgenic wheat genotype under 85 mM NaCl-salinity. Results showed that mtlD gene-contained transgenic plants had improved salt tolerance over non-transgenics, showing by better growth traits (i.e., number of leaves and leaf area per plant, root system size and plant dry weights), physio-biochemical attributes (i.e., levels of leaf chlorophylls, shoot free proline, total soluble sugars, soluble sugar fractions and mannitol, activities of enzymatic and non-enzymatic antioxidants, and contents of nutrient elements), yield (i.e., number of spikes and grain weight per plant, and 1000-grain weight) and yield quality (i.e., grain contents of starch, protein and soluble sugars). The mtlD gene transformation into wheat seeds appears to a better strategy to increase salt tolerance of plants through increased performance of mannitol and sugar accumulation, showing more of their salt stress-protecting role. The best performing mtlD transgenics could be incorporated in a breeding program to accumulate transgenes for stress tolerance in elite wheat genotypes in a step to commercialize these transgenics with the proper level of salt tolerance.
Keywords
Physio-Biochemical Attributes, Salt Stress, Transgenic Wheat, Yield and Its Quality
To cite this article
Mohamed Ahmed Seif El-Yazal, Hala Fawzi Eissa, Safia Mahmoud Abd El-Mageed Ahmed, Saad Mohamed Howladar, Safi-naz Sabet Zaki, Mostafa Mohamed Rady, The mtlD Gene-overexpressed Transgenic Wheat Tolerates Salt Stress Through Accumulation of Mannitol and Sugars, Plant. Vol. 4, No. 6, 2016, pp. 78-90. doi: 10.11648/j.plant.20160406.15
Copyright
Copyright © 2016 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Reference
[1]
Abebe, T., B. Guenzi, V. Martin, and J. Cushman. 2003. Tolerance of mannitol-accumulating transgenic wheat to water stress and salinity. Plant Physiol. 131:1748–1755.
[2]
Ahdam, I., F. Larher, and G.R. Stewart. 1979. Sorbitol, a compatible osmotic solute in Plantago maritime. New Phytol. 82:671–678.
[3]
Amiard, V., A. Morvan-Bertrand, J. P. Billard, C. Huault, F. Keller, and M. P. Prud’homme. 2003. Fructans, but not the sucrosyl-galactosides, raffinose and loliose, are affected by drought stress in perennial ryegrass. Plant Physiol. 132:2218–2229.
[4]
A. O. A. C. 1995. Official Methods of Analysis. 16th Edition. Association of Official Analytical Chemists, Washington, DC, USA.
[5]
Arnon, D. I. 1949. Copper enzymes in isolated chloroplasts. Polyphenol-oxidase in Beta vulgaris L. Plant Physiol. 24:1–5.
[6]
Askari, A., A. Pepoyan, and A. Parsaeimehr. 2012. Salt tolerance of genetic modified potato (Solanum tuberosum) cv. Agria by expression of a bacterial mtlD gene. Adv. Agric. Bot. 4:10–16.
[7]
Bates, L. S., R. P. Waldeen, and I. D. Teare. 1973. Rapid determination of free proline for water stress studies. Plant Soil. 39:205–207.
[8]
Beauchamp, C., and I. Fridovich. 1971. Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels. Anal. Biochem. 44:276–287.
[9]
Beers, R. F., and I. W. Sizer. 1952. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J. Biol. Chem. 195:133–140.
[10]
Bhauso, D. T. 2012. Transformation and characterization of transgenic groundnut (Arachis hypogaea L.) with mtlD gene for abiotic stress tolerance. Ph.D., Juangadh Agricultural University, Junagadh-362001, Gujarat, India.
[11]
Bhauso, T. D., R. Thankappan, A. Kumar, G. P. Mishra, J. R. Dobaria, and M. V. Rajam. 2014. Over-expression of bacterial mtlD gene confers enhanced tolerance to salt-stress and water-deficit stress in transgenic peanut (Arachis hypogaea) through accumulation of mannitol. Aust. J. Crop Sci. 8:413–421.
[12]
Bihmidine, S., C. T. Hunter, C. E. Johns, K. E. Koch, and D. M. Braun. 2013. Regulation of assimilate import into sink organs: update on molecular drivers of sink strength. Front. Plant Sci. 4:4–177.
[13]
Chapman, H. D., and P.F. Pratt. 1961. Methods of Analysis for Soil, Plants and Water. University of California, Division of Agricultural Science, Berkeley, CA, USA, pp. 56–63.
[14]
Chaturvedi, V., A. Bartiss, and B. Wong. 1997. Expression of bacterial mtlD in Saccharomyces cerevisiae results in mannitol synthesis and protects a glycerol-defective mutant from high-salt and oxidative stress. J. Bacteriol. 179:157–162.
[15]
Chiang, Y. J., C. Stushnoff, and A. E. McSay. 2005. Overexpression of mannitol-1-phosphate dehydrogenase increase mannitol accumulation and adds protection against chilling injury in petunia. J. Amer. Soc. Hortic. Sci. 130:605–610.
[16]
Christensen, A. H., R. A. Sharrock, and P. H. Quail. 1992. Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation. Plant Molec. Biol. 18:675–689.
[17]
Chunmei, H., Y. Aifang, Z. Weiwei, G. Qiang, and Z. Juren. 2010. Improved salt tolerance of transgenic wheat by introducing betA gene for glycine betaine synthesis. Plant Cell Tiss. Org. Cult. 101:65–78.
[18]
Conklin, P. L. 2001. Recent advances in the role and biosynthesis of ascorbic acid in plants. Plant Cell Environ. 24:383–394.
[19]
Dawson, C. R., and R. J. Magee. 1955. Ascorbic acid oxidase. In: Methods in Enzymology (Colowich, S.P., Kaplan, N.O., eds.), vol. 2, pp. 831–835, Academic press, New York, USA.
[20]
Foyer, C. H., M. Lelandais, and K.J. Kunert. 1994. Photooxidative stress in plants. Physiol. Plant. 92:696–717.
[21]
Galvani, A. 2007. The challenge of the food sufficiency through salt tolerant crops. Rev. Environ. Sci. Bio/Technol. 6:3–16.
[22]
Gao, Z., M. Sagi, and S. H. Lips. 1998. Carbohydrate metabolism in leaves and assimilate partitioning in fruits of tomato (Lycopersicon esculentumL.) as affected by salinity. Plant Sci. 135(2):149–159.
[23]
Granlund, M., and D. C. Zimmerman. 1975. Oil content of sunflower seed as determined by wide line Nuclear Megnetic Reasonance. Acad. Sci. 27:123–133.
[24]
Griffth, O. W. 1980. Determination of glutathione and glutathione disulfide using glutathione reductase and 2 vinyl pyridine. Anal. Biochem. 106:207–212.
[25]
Hayashi, F., T. Ichino, M. Osanai, and K. Wada. 2000. Oscillation and regulation of proline content by P5CS and ProDH gene expressions in the light/dark cycles in Arabidopsis thaliana L. Plant Cell Physiol. 41:1096–1101.
[26]
Helrich, K. 1990. Official Methods of Analysis. Vitamin C (Ascorbic Acid). 15th Edition. The Association of Official Analytical Chemists, Benjamin Franklin Station, Washington, DC, USA. pp. 1058–1059.
[27]
Hoque, M. A., M. N. Banu, E. Okuma, K. Amako, Y. Nakamura, Y. Shimoishi, and Y. Murata. 2007. Exogenous proline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities, and proline improves salt tolerance more than glycinebetaine in tobacco Bright Yellow-2 suspension-cultured cells. J. Plant Physiol. 164:1457–1468.
[28]
Hu, L., H. Lu, Q. Liu, X. Cheni, and X. Jiang. 2005. Overexpression of mtlD gene in transgenic Populus tomentosa improves salt tolerance through accumulation of mannitol. Tree Physiol. 25:1273–1281.
[29]
Husaini, A. M., and M. Z. Abdin. 2008. Overexpression of tobacco osmotin gene leads to salt stress tolerance in strawberry (Fragaria × ananassa Duch.) plants. Ind. J. Biotechnol. 7:465–471.
[30]
Irigoyen, J.J., D.W. Emerich, and M. Sanchez-Diaz. 1992. Water stress induced changes in the concentrations of proline and total soluble sugars in nodulated alfalfa (Medicago sativa) plants. Physiol. Plant. 8:455–460.
[31]
Jackson, M. L. 1967. Soil Chemical Analysis. Prentice Hall of India Pvt. Ltd., New Delhi, India, pp. 144–197; 326–338.
[32]
Kerepesi, I., and G. Galiba. 2000. Osmotic and salt stress-induced alteration in soluble carbohydrate content in wheat seedlings. Crop Sci. 40(2):482–487.
[33]
Keunen, E., D. Peshev, J. Vangronsveld, W. Van den Ende, and A. Cuypers. 2013. Plant sugars are crucial players in the oxidative challenge during abiotic stress: extending the traditional concept. Plant Cell Environ. 36:1242–1255.
[34]
Khare, N., D. Goyary, N.K. Singh, P. Shah, M. Rathore, S. Anandhan, D. Sharma, M. Arif, and Z. Ahmed. 2010. Transgenic tomato cv. Pusa Uphar expressing a bacterial mannitol-1-phosphate dehydrogenase gene confers abiotic stress tolerance. Plant Cell Tiss. Org. Cult. 103:267–277.
[35]
Kirk, R.S., and R. Sawyer. 1991. Composition and analysis of foods. 9th Edition, Longman Scientific Technical, pp. 182–235.
[36]
Kishor, P. B. K., Z. Hong, G. H. Miao, C. A. A. Hu, and D. P. S. Verma. 2005. Over production of 11-pyrroline-5-carboxylate synthetase increases proline production and confers osmotolerance in transgenic plants. Plant Physiol. 108:1387–1394.
[37]
Loewus, F. A. 1988. Ascorbic acid and its metabolic products. In: The biochemistry of plants (Preiss, J., ed.). Vol. 14. New York: Academic Press, pp. 85–107.
[38]
Maehly, A. C., and B. C. Chance. 1954. The assay of catalases and peroxidases. In: Methods of Biochemical Analysis (Glick, D., Ed.). Vol. 1, Interscience Publish. New York.
[39]
Mansour, M.M.F. 1998. Protection of plasma membrane of onion epidermal cells by glycinebetaine and proline against NaCl stress. Plant Physiol. Biochem. 36:767–772.
[40]
Masuda, R., K. Kaneko, and I. Yamashita. 1996. Sugar and cyclitol determination in vegetables by HPLC using postcolumn fluorescent derivatization. J. Food Sci. 61:1186–1190.
[41]
Matros, A., D. Peshev, M. Peukert, H.-P. Mock, and W. V. Ende. 2015. Sugars as hydroxyl radical scavengers: proof-of-concept by studying the fate of sucralose in Arabidopsis. Plant J. 82:822–839.
[42]
Munns, R., and M. Tester. 2008. Mechanisms of salinity tolerance. Ann. Rev. Plant Biol. 59:651–681.
[43]
Nanjo, T., M. Kobayashi, Y. Yoshiba, Y. Sanada, K. Wada, H. Tsukaya, Y. Kakubari, K. Yanaguchi-Shinozaki, and K. Shinozaki. 1999. Biological functions of proline in morphogenesis and osmotolerance revealed in antisense transgenic Arabidopsis thaliana. Plant J. 18:185–193.
[44]
Noctor, G., and C. H. Foyer. 1998. Ascorbate and glutathione: Keeping active oxygen under control. Ann. Rev. Plant Physiol. Plant Molec. Biol. 49:249–279.
[45]
Page, A. I., R. H. Miller, and D. R. Keeny. 1982. Methods of Soil Analysis. Part II. Chemical and Microbiological Methods. 2nd. Edition. Amer. Soc. Agron. Mad., WI, USA, pp. 225–246.
[46]
Peshev, D., R. Vergauwen, A. Moglia, E. Hideg, and W. Van den Ende. 2013. Towards understanding vacuolar antioxidant mechanisms: a role for fructans? J. Exp. Bot. 64:1025–1038.
[47]
Petrova, A. N., and T. T. Bolotina. 1956. Studies on the enzyme of starch metabolism in potato tubers during growth. Biochem. 21:4–15.
[48]
Pitman, M.G., and A. Lauchli. 2002. Global impact of salinity and agricultural ecosystems. In: Salinity: Environment - Plants - Molecules (Lauchli, A., Luttge, U., Eds.), pp. 3–20, Kluer, Netherland: Dordrecht.
[49]
Ramadan, A. M., H. F. Eissa, S. E. Hassanein, A. Z. Abdel Azeiz, O.M. Saleh, H.T. Mahfouz, F.M. El-Domyati, M. A. Madkour, and A. Bahieldin. 2013. Increased salt stress tolerance and modified sugar content of bread wheat stably expressing the mtlD gene. Life Sci. J. 10:2348–2362.
[50]
Rathinasabapathi, B. 2000. Metabolic engineering for stress tolerance: Installing osmoprotectant synthesis pathways–a review. Ann. Bot. 86:709–716.
[51]
Sairam, R. K., and G. C. Srivastava. 2002. Changes in antioxidant activity in sub-cellular fractions of tolerant and susceptible wheat genotypes in response to long term salt stress. Plant Sci. 162:897–904.
[52]
Sanchez, D. H., F. Lippold, H. Redestig, M. Hannah, A. Erban et al. 2008. Integrative functional genomics of salt acclimatization in the model legume Lotus japonicus. Plant J. 53, 973–987.
[53]
Schachtman, D. P., and R. Munns. 1992. Sodium accumulation in leaves of Triticum species that differ in salt tolerance. Aust. J. Plant physiol. 19:331–340.
[54]
Shen, B., R. G. Jensen, and H. J. Bohnert. 1997. Increased resistance to oxidative stress in transgenic plants by targeting mannitol biosynthesis to chloroplasts. Plant Physiol. 113:1177–1183.
[55]
Sivamani, E., A. Bahieldin, J. M. Wraith, T. Al-Niemi, W. E. Dyer, T. D. Ho, and R. Qu. 2000. Improved biomass productivity and water use efficiency under water deficit conditions in transgenic wheat constitutively expressing the barley HVA1 gene. Plant Sci. 155:1–9.
[56]
Sivkumar, P., P. Sharmila, and P. Pardha-Saradhi. 2000. Proline alleviates salt stress-induced enhancement in ribulose-l,5-bisphosphate oxygenase activity. Biochem. Biophys. Res. Commun. 279:512–515.
[57]
Snedecor, G. W., and W. G. Cochran. 1990. "Statistical Methods". 8th Ed. Iowa State Univ. Press Ames, Iowa, USA, p. 609.
[58]
Southern, E. M. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Molec. Biol. 98:503–517.
[59]
Srivastava, A. K., N. K. Ramaswamy, P. Suprasanna, and S.F. D’Souza. 2010. Genome-wide analysis of thiourea-modulated salinity stress responsive transcripts in seeds of Brassica juncea: identification of signalling and effector components of stress tolerance. Ann. Bot. 106:663–674.
[60]
Stoop, J. M. H., J. D. Williamson, and D. M. Pharr. 1996. Mannitol metabolism in plants: a method for coping with stress. Trends Plant Sci. 1:139–144.
[61]
Su, J., P. L. Chen, and R. Wu. 1999. Transgene expression of mannitol-1-phosphate dehydrogenase enhanced the salt stress tolerance of the transgenic rice seedlings. Sci. Agric. Sin. 32:101–103.
[62]
Taneja, S. R., and R. C. Sachar. 1974. Introduction of polyphenol oxidase in germinating wheat seeds. Phytochem. 13:2695–2702.
[63]
Tarkowski, L.P., and W. Van den Ende. 2015. Cold tolerance triggered by soluble sugars: a multifaceted countermeasure. Fronti. Plant Sci. 6:1–7.
[64]
Thomas, J. C., R. D. Armond, and H. J. Bohnert. 1992. Influnce of NaCl on growth, proline and phosphoenolpyruvate carboxylase levels in Mesembryanthemum crystallinum suspension cultures. Plant Physiol. 98:626–631.
[65]
Thomas, J. C., M. Sepahi, B. Arendall, and H. J. Bohnert. 1996. Enhancement of seed germination in high salinity by engineering mannitol expression in Arabidopsis thaliana. Plant Cell Environ. 18:801–806.
[66]
Van den Ende, W., and S. El-Esawe. 2014. Sucrose signaling pathways leading to fructan and anthocyanin accumulation: a dual function in abiotic and biotic stress responses? Environ. Exp. Bot. 108:4–13.
[67]
Van den Ende, W., and R. Valluru. 2009. Sucrose, sucrosyl oligosaccharides, and oxidative stress: scavenging and salvaging? J. Exp. Bot. 60:9–18.
[68]
Weeks, J. T., O. D. Anderson, and A. E. Blechl. 1993. Rapid production of multiple independent lines of fertile transgenic wheat (Triticum aestivum). Plant Physiol. 102:1077–1084.
Browse journals by subject