Biologia plantarum 54:631-638, 2010 | DOI: 10.1007/s10535-010-0113-0

Functional characterization of the apple MdSAMDC2 gene by ectopic promoter analysis and over-expression in tobacco

L. -L. Zhao1,2,3, L. -Q. Song3, C. -X. You1,2, T. Moriguchi4, Y. -J. Hao1,2,*
1 State Key Laboratory of Crop Biology, Shandong Agricultural University, Tai-An, Shandong, P.R. China
2 College of Hort. Sci. and Engineering, Shandong Agricultural University, Tai-An, Shandong, P.R. China
3 Yan-Tai Agricultural Academy, Yan-Tai, Shandong, P.R. China
4 National Institute of Fruit Tree Science, Tsukuba, Ibaraki, Japan

The expression of MdSAMDC2 gene, which encodes S-adenosylmethionine decarboxylase (SAMDC) in apple, was up-regulated by low temperature, salt and drought stresses. To identify its in vivo biological functions in the responses to stresses, the promoter region of MdSAMDC2 was isolated and characterized by analyzing the cis-acting regulatory elements and GUS reporter gene by an Agrobacterium-mediated transformation. GUS activity was enhanced upon salt and cold stresses, indicating that MdSAMDC2 promoter region controls gene transcription under stresses. In parallel, several lines of the transgenic tobacco plants over-expressing MdSAMDC2 were obtained. The contents of three polyamines greatly increased in the transgenic lines compared with the non-transgenic WT control. Upon exposed to low temperature (4 °C), salt (150 and 250 mM NaCl) and osmotic (20 % polyethylene glycol) stresses, transgenic plants produced more free polyamines and more active antioxidative enzymes such as superoxide dismutase and catalase than the WT control. Meanwhile, malondialdehyde content, an indicator for membrane lipid peroxidation, decreased in transgenic plants relative to the WT control. Thus, over-expression of MdSAMDC2 in tobacco conferred tolerance to stresses.

Keywords: antioxidative enzymes; lipid peroxidation; low temperature; Nicotiana tabacum; osmotic stress; salinity
Subjects: Agrobacterium tumefaciens; apple; catalase; β-glucuronidase; malondialdehyde; Malus domestica; Nicotiana tabacum; nucleotide sequences; osmotic stress; polyamines; polyethylene glycol; proline; salinity; superoxide dismutase (SOD); temperature low; tobacco; transgenic plants

Received: April 18, 2009; Accepted: October 11, 2009; Published: December 1, 2010Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Zhao, L.-L., Song, L.-Q., You, C.-X., Moriguchi, T., & Hao, Y.-J. (2010). Functional characterization of the apple MdSAMDC2 gene by ectopic promoter analysis and over-expression in tobacco. Biologia plantarum54(4), 631-638. doi: 10.1007/s10535-010-0113-0.
Download citation

References

  1. Bolle, C., Herrmann, R.G., Oelmller, R.: A spinach cDNA with homology to S-adenosylmethionine decarboxylase. - Plant Physiol. 107: 1461-1462, 1995. Go to original source...
  2. Bradford, M.M.: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. - Anal. Biochem. 72: 248-254, 1976. Go to original source...
  3. Capell, T., Bassie, L., Christou, P.: Modulation of the polyamine biosynthetic pathway in transgenic rice confers tolerance to drought stress. - Proc. nat. Acad. Sci. USA 101: 9909-9914, 2004. Go to original source...
  4. Chinnusamy, V., Ohta, M., Kanrar, S., Lee, B., Hong, X., Agarwal, M., Zhu, J.K.: ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. - Genes Dev. 17: 1043-1054, 2003. Go to original source...
  5. Cowley, T., Walters, D.R.: Polyamine metabolism in barley reacting hypersensitively to the powdery mildew fungus Blumeria graminis f. sp. hordei. - Plant Cell Environ. 25: 461-468, 2002. Go to original source...
  6. De Agazio, M., Zacchini, M., Federico, R., Grego, S.: Putrescine accumulation in maize roots treated with spermidine: evidence for spermidine to putrescine conversion. - Plant Sci. 111: 181-185, 1995. Go to original source...
  7. Dresselhaus, T., Barcelo, P., Hagel, C., Lörz, H., Humbeck, K.: Isolation and characterization of a Tritordeum cDNA encoding S-adenosylmethionine decarboxylase that is circadian-clock regulated. - Plant mol. Biol. 30: 1021-1033, 1996. Go to original source...
  8. Duhaze, C., Gouzerh, G., Gagneul, D., Larher, F., Bouchereau, A.: The conversion of spermidine to putrescine and 1,3-diaminopropane in the roots of Limonium tataricum. - Plant Sci. 163: 39-646, 2002. Go to original source...
  9. Evans, P.T., Malmberg, R.L.: Do polyamines have roles in plant development? - Annu. Rev. Plant Physiol. Plant mol. Biol. 40: 235-269, 1989. Go to original source...
  10. Franceschetti, M., Hanfrey, C., Scaramagli, S., Torrigiani, P., Bagni, N., Burtin, D., Michael, A.J.: Characterization of monocot and dicot plant S-adenosyl-l-methionine decarboxylase gene families including identification in the mRNA of a highly conserved pair of upstream overlapping open reading frames. - Biochem. J. 353: 403-409, 2001. Go to original source...
  11. Hao, Y.J., Zhang, Z.L., Kitashiba, H., Honda, C., Ubi, B., Kita, M., Moriguchi, T.: Molecular cloning and functional characterization of two apple S-adenosylmethionine decarboxylase genes and their different expression in fruit development, cell growth and stress responses. - Gene 350: 41-50, 2005. Go to original source...
  12. He, L., Ban, Y., Inoue, H., Matsuda, N., Liu, J., Moriguchi, T.: Enhancement of spermidine content and antioxidant capacity in transgenic pear shoots overexpressing apple spermidine synthase in response to salinity and hyperosmosis. - Phytochemistry 69: 2133-2141, 2008. Go to original source...
  13. Jefferson, R.A., Kavanagh, T.A., Bevan, M.W.: GUS-fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. - EMBO J. 6: 3901-3907, 1987. Go to original source...
  14. Kashiwagi, K., Taneja, S.K., Liu, T.Y., Tabor, C.W., Tabor, H.: Spermidine biosynthesis in Saccharomyces cerevisiae. Biosynthesis and processing of a proenzyme form of S-adenosylmethionine decarboxylase. - J. biol. Chem. 265: 22321-22328, 1990.
  15. Kumar, A., Taylor, M.A., Mad Arif, S.A., Davies, H.: Potato plants expressing antisense and sense S-adenosylmethionine decarboxylase (SAMDC) transgenes show altered levels of polyamines and ethylene: antisense plants display abnormal phenotypes. - Plant J. 9: 147-158, 1996. Go to original source...
  16. Kwon, S.W., Ko, B.R., Bai, D.G.: Changes in antioxidant enzymes and polyamines in response to low temprature chilling in watermelon. - Acta Hort. 620: 111-117, 2003. Go to original source...
  17. Lee, M.M., Lee, S.H., Park, K.Y.: Characterization and expression of two members of the S-adenosylmethionine decarboxylase gene family in carnation flower. - Plant mol. Biol. 34: 371-382, 1997. Go to original source...
  18. Li, Z.Y., Chan, S.Y.: Differential accumulation of the S-adenosylmethionine decarboxylase transcript in rice seedlings in response to salt and drought stresses. - Theor. appl. Genet. 100: 782-788, 2000. Go to original source...
  19. Luo, K., Zheng, X., Chen, Y., Xiao, Y., Zhao, D., McAvoy, R., Pei, Y., Li, Y.: The maize Knotted1 gene is an effective positive selectable marker gene for Agrobacterium-mediated tobacco transformation. - Plant Cell Rep. 25: 403-409, 2006. Go to original source...
  20. Mad Arif, S.A., Taylor, M.A., George, L.A., Butler, A.R., Burch, L.R., Davies, H.V., Stark, M.J.R., Kumar, A.: Characterisation of the S-adenosylmethionine decarboxylase (SAMDC) gene in potato. - Plant mol. Biol. 26: 327-338, 1994. Go to original source...
  21. Marco, F., Carrasco, P.: Expression of the pea S-adenosylmethionine decarboxylase is involved in developmental and environmental responses. - Planta 214: 641-647, 2002. Go to original source...
  22. Mehta, R.A., Cassol, T., Li, N., Ali, N., Handa, A.K., Mattoo, A.K.: Engineered polyamine accumulation in tomato enhances phytonutrient content, juice quality, and vine life. - Nat. Biotechnol. 20: 613-618, 2002. Go to original source...
  23. Meshi, T., Iwabuchi, M.: Plant transcription factors. - Plant Cell Physiol. 36: 1405-1420, 1995.
  24. Mo, H., Pua, E.C.: Up-regulation of arginine decarboxylase gene expression and accumulation of polyamines in mustard (Brassica juncea) in response to stress. - Physiol. Plant. 114: 439-449, 2002. Go to original source...
  25. Pajunen, A., Crozat, A., Janne, O.A., Ihalainen, R., Laitinen, P.H., Stanley, B., Madhubala, R., Pegg, A.E.: Structure and regulation of mammalian S-adenosylmethionine decarboxylase. - J. biol. Chem. 263: 17040-17049, 1988.
  26. Pérez-Amador, M.A., Leon, J., Green, P.J., Carbonell, J.: Induction of the arginine decarboxylase ADC2 gene provides evidence for the involvement of polyamines in the wound response in Arabidopsis. - Plant Physiol. 130: 1454-1463, 2002. Go to original source...
  27. Pillai, M.A., Akiyama, T.: Differential expression of an S-adenosylmethionine decarboxylase gene involved in polyamine biosynthesis under low temperature stress in japonica and indica rice genotypes. - Mol. Genet. Genomics 271: 141-149, 2004. Go to original source...
  28. Porebski, S.L., Bailey, G., Baum, B.R.: Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. - Plant mol. Biol. Rep. 15: 8-15, 1997. Go to original source...
  29. Roy, M., Wu, R.: Overexpression of S-adenosylmethionine decarboxylase gene in rice increases polyamine level and enhances sodium chloride-stress tolerance. - Plant Sci. 163: 987-992, 2002. Go to original source...
  30. Shen, W.Y., Kazuyoshi, N., Shoji, T.: Involvement of polyamines in the chilling tolerance of cucumber cultivars. - Plant Physiol. 124: 431-440, 2000. Go to original source...
  31. Simpson, S.D., Nakashima, K., Narusaka, Y., Seki, M., Shinozaki, K., Yamaguchi-Shinozaki, K.: Two different novel cis-acting elements of erd1, a clpA homologous Arabidopsis gene function in induction by dehydration stress and dark-induced senescence. - Plant J. 33: 259-270, 2003. Go to original source...
  32. Sofo, A., Dichio, B., Xiloyannis, C., Masia, A.: Effects of different irradiance levels on some antioxidant enzymes and on malondialdehyde content during rewatering in olive tree. - Plant Sci. 66: 293-302, 2004. Go to original source...
  33. Su, G.X., Bai, X.: Contribution of putrescine degradation to proline accumulation in soybean leaves under salinity. - Biol. Plant. 52: 796-799, 2008. Go to original source...
  34. Thu-Hang, P., Bassie, L., Safwat, G., Treng-Nghia, P., Christou, P., Capell, T.: Expression of a heterologous S-adenosylmethionine decarboxylase cDNA in plants demonstrates that changes in S-adenosyl-L-methionine decarboxylase activity determine levels of the higher polyamines spermidine and spermine. - Plant Physiol. 129: 1744-1754, 2002. Go to original source...
  35. Tian, A.G., Zhao, J.Y., Zhang, J.S., Gai, J.Y., Chen, S.Y.: Genomic characterization of the S-adenosylmethionine decarboxylase genes from soybean. - Theor. appl. Genet. 108: 842-850, 2004. Go to original source...
  36. Tiburcio, A.F., Campos, J.L., Figueras, X., Besford, R.T.: Recent advances in the understanding of polyamine functions during plant development. - Plant Growth Regul. 12: 331-340, 1993. Go to original source...
  37. Tonon, G., Kevers, C., Faivre-Rampant, O., Graziani, M., Gaspar, T.: Effect of NaCl and mannitol iso-osmotic stresses on proline and free polyamine levels in embryogenic Fraxinus angustifolia callus. - J. Plant Physiol. 161: 701-708, 2004. Go to original source...
  38. Valliyodan, B., Nguyen, H.T.: Understanding regulatory networks and engineering for enhanced drought tolerance in plants. - Curr. Opin. Plant Biotechnol. 9: 189-195, 2006.
  39. Vinocur, B., Altman, A.: Recent advances in engineering plant tolerance to abiotic stress: achievements and limitations. - Curr. Opin. Biotechnol. 16: 123-132, 2005. Go to original source...
  40. Waie, B., Rajam, M.V.: Effect of increased polyamine biosynthesis on stress responses in transgenic tobacco by introduction of human S-adenosylmethionine gene. - Plant Sci. 164: 727-734, 2003. Go to original source...
  41. Watson, M.B., Malmberg, R.L.: Regulation of Arabidopsis thaliana (L.) Heynh arginine decarboxylase by potassium deficiency stress. - Plant Physiol. 111: 1077-1083, 1996. Go to original source...
  42. Wen, X.P., Pang, X.M., Matsuda, N., Kita, M., Inoue, H., Hao, Y.J., Honda, C., Moriguchi, T.: Over-expression of the apple spermidine synthase gene in pear confers multiple abiotic stress tolerance by altering polyamine titers. - Transgenic Res. 17: 251-263, 2008. Go to original source...
  43. Wi, S.J., Kim, W.T., Park, K.Y.: Overexpression of carnation S-adenosylmethionine decarboxylase gene generates a broad-spectrum tolerance to abiotic stresses in transgenic tobacco plants. - Plant Cell Rep. 25: 1111-1121, 2006. Go to original source...
  44. Wu, A.M., Liu, J.Y.: An improved method of genomic walking for promoter sequences cloning. - Chin. J. Biochem. mol. Biol. 22: 243-246, 2006.
  45. Zhang, J., Kirkham, M.B.: Drought-stress-induced changes in activities of superoxide dismutase, catalase, and peroxidase in wheat species. - Plant Cell Physiol. 35: 785-791, 1994. Go to original source...