Enhance salinity resistance in Arabidopsis thaliana by overexpression of phosphoethanolamine N-methyltransferase gene from spinach (Spinacia oleracea, Iranian landrace)

Allahi, Somayeh; Sohani, Mohammad; Hasani, Hassan

Document Type : Research Paper

Authors

¹ Ph.D. Student, Department of Agricultural Biotechnology, Faculty of Agriculture, Guilan University, Rasht, Iran

² Associate Professor, Department of Agricultural Biotechnology, Faculty of Agriculture Science, Guilan University, Rasht, Iran

³ Assistant Professor, Department of Agricultural Biotechnology, Faculty of Agriculture Science, Guilan University, Rasht, Iran.

Abstract

One of the plant resistant mechanisms to abiotic stresses is production of a compatible solute named glycine betaine. Choline is the precursor of this important metabolite and it is also essential compound for the structural integrity and signaling of cell membrane. In plants, the most important step of choline production is catalyzed by cytoplasmic phosphoethanolamine N-methyltransferase (PEAMT ; EC 2.1.1.103) enzyme. In this study, PEAMT gene from spinach (Spinacia oleracea Iranian landrace) was amplified using specific primers and cloned into an intermediate cloning vector (pJET). In order to overexpression of PEAMT gene, the construct of PBI121GUS-9:PEAMT was made and finally was transferred to the Agrobacterium tumefaciens GV3101 (PMP90). Floral dip method was used for transformation and initial analysis of putative transgenic plants was tested in selective medium containing kanamycin. Then resistant seedlings at the molecular level were evaluated using PCR and RT-PCR methods. Results confirmed plant transformation in the level of transcription. Subsequently, The phenotypic analysis under salt stress showed that the main root length of transgenic plants was significantly longer than control nontransgenics. In adition, glycinebetaine contents and peroxidase activity were significantly higher in transgenic compare to non-transgenics control plants.

Keywords

20.1001.1.22520783.1395.6.15.4.0

Main Subjects

Biotic and Abiotic stress

References

Amako K, Chen GX, Asada K (1994) Separate assays specific for ascorbate peroxidase and guaiacol peroxidase and for the chloroplastic and cytosolic isozymes of ascorbate peroxidase in plants. Plant Cell Physiol. 35(3): 497-504.

Bobenchik AM, Augagneur Y, Hao B, Hoch JC, Mamoun CB (2011) Phosphoethanolamine methyltransferases in phosphocholine biosynthesis: functions and potential for antiparasite therapy. FEMS Microbiol Rev. 35(4): 609-619.

Chen TH, Murata N (2011) Glycine betaine protects plants against abiotic stress: mechanisms and biotechnological applications. Plant Cell Environ. 34(1): 1-20.

Cruz-Ramírez A, López-Bucio J, Ramírez-Pimentel G, Zurita-Silva A, Sánchez-Calderon L, Ramírez-Chávez E, González-Ortega E, Herrera-Estrella L (2004) The xipotl mutant of Arabidopsis reveals a critical role for phospholipid metabolism in root system development and epidermal cell integrity. Plant Cell. 16(8): 2020-2034.

Einset J, Connolly EL (2009) Glycine betaine enhances extracellular processes blocking ROS signaling during stress. Plant Signal Behav. 4(3): 197-199.

Esfahani K, Salmanian AH (2013) Designing and construction of new plant expression vectors with more recognition sites of restriction enzymes. 8^th National Congress of Biotechnology, Tehran, Iran.

Galvan-Ampudia CS, Testerink C (2011) Salt stress signals shape the plant root. Curr Opin Plant Biol. 14(3): 296-302.

Grieve CM, Grattan SR (1983) Rapid assay for determination of water-soluble quaternary ammonium-compounds. Plant Soil. 70(2): 303-30.

Kaeppler SM, Kaeppler HF, Rhee Y (2000) Epigenetic aspects of somaclonal variation in plants. Plant Mol. Biol. 43(2-3): 179-188.

Kumar S, Dhingra A, Daniell H (2004) Plastid-expressed betaine aldehyde dehydrogenase gene in carrot cultured cells, roots, and leaves confers enhanced salt tolerance. Plant Physiol. 136(1): 2843-2854.

Li F, Guo S, Zhao Y, Chen D, Chong K, Xu Y (2010) Overexpression of a homopeptide repeat-containing bHLH protein gene (OrbHLH001) from Dongxiang Wild Rice confers freezing and salt tolerance in transgenic Arabidopsis. Plant Cell Rep. 29(9): 977-986.

McNeil SD, Nuccio ML, Ziemak MJ, Hanson AD (2001) Enhanced synthesis of choline and glycine betaine in transgenic tobacco plants that overexpress phosphoethanolamine N-methyltransferase. Proc. Natl. Acad Sci. USA. 98(17): 10001-10005.

Mou Z, Wang X, Fu Z, Dai Y, Han C, Ouyang J, Bao F, Hu Y, Li J (2002) Silencing of phosphoethanolamine N-methyltransferase results in temperature-sensitive male sterility and salt hypersensitivity in Arabidopsis. Plant Cell. 14(9): 2031-2043.

Nuccio ML, Ziemak MJ, Henry SA, Weretilnyk EA, Hanson AD (2000) cDNA Cloning of Phosphoethanolamine N-Methyltransferase from Spinach by Complementation in Schizosaccharomyces pombe and Characterization of the Recombinant Enzyme. J. Biol. Chem. 275(19): 14095-14101.

Rhodes D, Hanson A (1993) Quaternary ammonium and tertiary sulfonium compounds in higher plants. Annu. Rev. Plant Biol. 44(1): 357-384.

Sakamoto A, Valverde R, Chen TH, Murata N (2000) Transformation of Arabidopsis with the codA gene for choline oxidase enhances freezing tolerance of plants. Plant J. 22(5): 449-453.

Sakr M, El-Sarkassy NM, Fuller MP (2012). Osmoregulators proline and glycine betaine counteract salinity stress in canola. . Agron. Sustain. Dev. 32(3): 747-754.

Smith DD, Summers PS, Weretilnyk EA (2000) Phosphocholine synthesis in spinach: Characterization of phosphoethanolamine N-methyltransferase. Physiol. Plantarum. 108(3): 286-294.

Su J, Hirji R, Zhang L, He C, Selvaraj G, Wu R (2006) Evaluation of the stress-inducible production of choline oxidase in transgenic rice as a strategy for producing the stressprotectant glycinebetaine. J. Exp. Bot. 57(5): 1129-1135.

Wu S, Yu Z, Wang F, Li W, Ye C, Li J, Tang J, Ding J, Zhao J, Wang B (2007) Cloning, characterization, and transformation of the phosphoethanolamine N-methyltransferase gene (ZmPEAMT1) in maize (Zea mays L.). Mol. Biotechnol. 36(2): 102-112.

Ye C, Ye C, Wu S, Ye C, Wu S, Yang Q, Ye C, Wu S, Yang Q, Ma C, Yang G, Wang B (2005) Cloning, sequencing and salt induced expression of PEAMT and BADH in oilseed rape (Brassica napus). DNA Seq. 16(5): 364-371.

Zeisel SH (2006) Choline: critical role during fetal development and dietary requirements in adults. Ann. Rev. Nutr. 26: 229-250.

Zhang H, Murzello C, Sun Y, Kim MS, Xie X, Jeter RM, Zak JC, Dowd SE, Paré PW (2010) Choline and osmotic-stress tolerance induced in Arabidopsis by the soil microbe Bacillus subtilis (GB03). Mol. Plant Microbe. Interact. 23(8): 1097-1104.

Zhang J (2003) Evolution by gene duplication: an update. Trends Ecol. Evol. 18(6): 292-298.

Zhang J, Tan W, Yang XH, Zhang HX (2008) Plastidexpressed choline monooxygenase gene improved salt and drought tolerance through accumulation of glycine betaine in tobacco. Plant Cell Rep. 27(6): 1113-1124.

Zhang X, Henriques R, Lin SS, Niu QW, Chua NH (2006) Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method. Nat. Protoc. 1(2): 641-646.

Zhou S, Chen X, Zhang X, Li Y (2008) Improved salt tolerance in tobacco plants by co-transformation of a betaine synthesis gene BADH and a vacuolar Na+/H+ antiporter gene SeNHX1. Biotechnol. Lett. 3(2) 369- 376.

Crop Biotechnology

Enhance salinity resistance in Arabidopsis thaliana by overexpression of phosphoethanolamine N-methyltransferase gene from spinach (Spinacia oleracea, Iranian landrace)

References

References

Volume 6, Issue 15 - Serial Number 15
December 2016
Pages 37-52

Enhance salinity resistance in Arabidopsis thaliana by overexpression of phosphoethanolamine N-methyltransferase gene from spinach (Spinacia oleracea, Iranian landrace)

References

References

Volume 6, Issue 15 - Serial Number 15December 2016Pages 37-52

Volume 6, Issue 15 - Serial Number 15
December 2016
Pages 37-52