Isolation, molecular cloning and expression analysis of Aeluropus littoralis Monodehydroascorbate reductase (MDHAR) gene under salt stress

Document Type : Original research paper

Authors

1 Department of Plant Breeding, Sari Agricultural Sciences and Natural Resources University, Sari, Iran.

2 Genetics and Agricultural Biotechnology Institute of Tabarestan, Sari Agricultural Sciences and Natural Resources University, Sari, Iran.

Abstract

Plants are naturally exposed to frequent changes in environmental conditions such as salt stress affecting their growth and development, and consequently a considerable reduction in total productivity. Monodehydroascorbate reductase (MDHAR), a key enzyme in ascorbate-glutathione cycle, serves as an important antioxidative enzyme in scavenging of reactive oxygen spices (ROS). In this study, MDHAR gene from Aeluropus littoralis was isolated using specific primers by RT-PCR and its expression variation was investigated at 0, 150, 300, 450 and 600 mM of NaCl through Real-time PCR. The results indicated that A. littoralis MDHAR gene compromised 1436 bp without any introns, showing a high similarity with Sorghum bicolor. The only alteration of MDHAR expression was observed at 300 mM NaCl, i.e. cytosolic MDHAR in both shoot and root was increased 1.3 and 1.4 fold compared to control samples. It could be concluded that accumulation of MDHAR reduces at Nacl concentrations higher than 300 mM. 

Keywords

- References
[1]        Ashraf, M., (2009). Biotechnological approach of improving plant salt tolerance using antioxidants as markers. Biotechnol Adv, 27: 84–93.
[2]        Eltayeb, A. E., Kawano, N., Badawi, G. H., Kaminaka, H., Sanekata, T., Morishima, I., Murishima, T., Shibahara, S., Inanaga & Tanaka, K. (2006). Enhanced tolerance to ozone and drought stresses in transgenic tobacco overexpressing dehydroascorbate reductase in cytosol. Physiol Plant, 127: 57-65.
[3]        Eltayeb, A. E., Kawano, N., Badawi, G. H., Kaminaka, H., Sanekata, T., Shibahara, S., Inanaga, and K., Tanaka & Tanaka, K. (2007). Overexpression of monodehydroascorbate reductase in transgenic tobacco confers enhanced tolerance to ozone, salt and polyethylene glycol stresses. Planta, 225(5): 1255-1264.
[4]        Eltayeb, H.A., Badejo, A.A., Fujikawa, Y., Esaka, M. (2011). Gene expression of monodehydroascorbate reductase and dehydroascorbate reductase during fruit ripening and in response to environmental stresses in acerola (Malpighia glabra). J Plant Physiol, 168: 619–627.
[5]        Foyer, C., Halliwell, B., (1976). The presence of glutathione and glutathione reductase in chloroplasts: a proposed role in ascorbic acid metabolism. Planta, 133: 21–25.
[6]        Foyer, C.H., & Mullineaux, P. (1994). Causes of photooxidative stress and amelioration of defense systems in plants. CRC Press Inc.
[7]        Gautier, H., Lopez-Lauri, F., Massot, C., Murshed, R., Marty, I., Grasselly, D., Keller, C., Sallanon, H., Genard. (2010). Impact of ripening and salinity on tomato fruit ascorbate content and enzymatic activities related to ascorbate recycling. Funct Plant Sci Biotechnol, 4: 66–75.
[8]        Grantz A, D.A. Brummell, A.B. Bennett. (1995). Ascorbate free-radical reductase messenger-RNA levels are induced by wounding. Plant Physiol, 108: 411-418.
[9]        Gulzar, S., Khan, M. A., & Ungar, I. A. (2003). Salt tolerance of a coastal salt marsh grass. Commun Soil Sci Plant Anal, 34: 2595-2605.
[10]      Haroldsen, V.M., Chi-Ham, C.L., Kulkarni, S., Lorence, A., Bennett, A.B., (2011). Constitutively expressed DHAR and MDHAR influence fruit, but notfoliar ascorbate levels in tomato. Plant Physiol Biochem, 49: 1244–1249.
[11]      Kavitha K., S. George, G. Venkataraman, A. Parida. (2010). Salt-inducible chloroplastic monodehydroascorbate reductase from halophyte Avicennia marina confers salt stress tolerance on transgenic plants. Biochim, 92: 1321e1329.
[12]      Kaya, C., Kirnak, H., Higgs, D., Saltali, K. (2002). Supplementary calcium enhances plant growth and fruit yield in strawberry cultivars grown at high (NaCl) salinity. Sci Hortic, 93: 65–74.
[13]      Li, F., Wu, Q.Y., Sun, Y.L., Wang, L.Y., Yang, X.H., Meng, Q.W. (2010). Overexpression of chloroplastic monodehydroascorbate reductase enhanced tolerance to temperature and methyl viologen-mediated oxidative stresses. PhysiolPlant, 139: 421–434.
[14]      Li, M.Y., Liu, Y.J., (1994). Halophytes of Yellow River Delta in north Shandong Province of China. J Qufu Normal Univ, 125–133.
[15]      Lunde C, Baumann U, Shirley NJ, Drew DP, Fincher GB. (2006). Gene structure and expression pattern analysis of three monodehydroascorbate reductase (Mdhar) genes in Physcomitrella patens: implications for the evolution of the MDHAR family in plants. Plant Mol Biol, 60: 2590–675.
[16]      Naliwajski, M.R., Sklodowska, M. (2014). The oxidative stress and Antioxidant systems in cucumber cells during acclimation to salinity. Biol Plant, 58: 47–54.
[17]      Qin, A., Shi, Q., & Yu, X. (2011). Ascorbic acid contents in transgenic potato plants overexpressing two dehydroascorbate reductase genes. MolBiol Rep, 38(3): 1557-1566.
[18]      Sen, A., Alikamanoglu, S. (2013). Antioxidant enzyme activities, malondialdehyde,and total phenolic content of PEG-induced hyperhydric leaves in sugar beet tissue culture. In Vitro Cell Dev Biol Plant, 49: 396–404.
[19]      Sultana, S., Khew, C. Y., Morshed, M. M., Namasivayam, P., Napis, S., & Ho, C. L. (2012). Overexpression of monodehydroascorbate reductase from a mangrove plant (AeMDHAR) confers salt tolerance on rice. J Plant Physiol, 169(3): 311-318.
[20]      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.
[21]      Yabuta, Y., Maruta, T., Yoshimura, K., Ishikawa, T., Shigeoka, S. (2004). Two distinct redox signaling pathways for cytosolic APX induction under photooxidative stress. Plant Cell Physiol, 45: 1586–1594.
[22]      Yoon, H.S., Lee, H., Lee, I.A., Kim, K.Y. and Jo, J.K. (2004). Molecular cloning of the monodehydro-ascorbate reductase gene from Brassica campestris and analysis of its mRNA level in response to oxidative stress. Biochim Biophys Acta, 1658: 181–186.
[23]      Zouari, N., Ben Saad, R., Legavre, T., Azaza, J., Sabau, X., Jaoua, M., Masmoudi, K., Hassairi, A. (2007). Identification and sequencing of ESTs from the halophyte grass Aeluropus littoralis. Genet, 404: 61–69.
[24]      Zhu, J.K. (2002). Salt and drought stress signal transduction in plants. Annu Rev Plant Biol, 53: 247–273.
 
 
 
 
 
 
Volume 3, Issue 1
June 2015
Pages 72-80
  • Receive Date: 26 May 2015
  • Revise Date: 26 August 2015
  • Accept Date: 26 August 2015
  • First Publish Date: 26 August 2015