Biologia Plantarum

, Volume 56, Issue 2, pp 337–343 | Cite as

The effect of water deficit and excess copper on proline metabolism in Nicotiana benthamiana

Original Papers
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Abstract

Fluctuation in proline content is a widespread phenomenon among plants in response to heavy metal stress. To distinguish between the participation of water deficit and copper on changes in proline metabolism, potted plants and floating leaf discs of tobacco were subjected to CuSO4 treatments. The application of copper increased the proline content in the leaves concomitantly with decreased leaf relative water content and increased abscisic acid (ABA) content in the potted plant. Excess copper increased the expression of two proline synthesis genes, pyrroline-5-carboxylate synthetase (P5CS) and ornithine aminotransferase (OAT) and suppressed proline catabolism gene, proline dehydrogenase (PDH). However, in the experiment with tobacco leaf discs floating on CuSO4 solutions, the excess copper decreased proline content and suppressed the expression of the P5CS, OAT and PDH genes. Therefore, proline accumulation in the potted tobacco plants treated with excess Cu treatment might not be the consequence of the increased copper content in tobacco leaves but rather by the accompanied decrease in water content and/or increased ABA content.

Additional key words

abscisic acid gene expression tobacco 

Abbreviations

ABA

abscisic acid

OAT

ornithine aminotransferase

P5CS

pyrroline-5-carboxylate synthetase

PDH

proline dehydrogenase

RWC

relative water content

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Notes

Acknowledgements

We are grateful to Dr. C.-J. Chang, A. Frary and V. Panwar for critically reviewing and editing the manuscript.

References

  1. Armengaud, P., Thiery, L., Buhot, N., March, G.G., Savoure, A.: Transcriptional regulation of proline biosynthesis in Medicago truncatula reveals developmental and environmental specific features. — Physiol. Plant. 120: 442–450, 2004.PubMedCrossRefGoogle Scholar
  2. Bassi, R., Sharma, S.S.: Proline accumulation in wheat seedlings exposed to zinc and copper. — Phytochemistry 33: 1339–1342, 1993.CrossRefGoogle Scholar
  3. Bates, L.S., Waldren, R.P., Teare, I.D.: Rapid determination of free proline for water-stress studies. — Plant Soil 39: 205–207, 1973.CrossRefGoogle Scholar
  4. Bradford, M.M.: A rapid and sensitive method for the quantitation of microgram quantities for protein utilizing the principles of protein-dye binding. — Anal. Biochem. 72: 248–254, 1976.PubMedCrossRefGoogle Scholar
  5. Charest, C., Phan, C.T.: Cold accumulation of wheat (Triticum aestvum): properities of enzymes involved in proline metabolism. — Physiol. Plant. 80: 159–168, 1990.CrossRefGoogle Scholar
  6. Canas, R.A., Villalobos, D.P., Diaz-Moreno, S.M., Canovas, F.M., Canton, F.R.: Molecular and functional analyses support a role of ornithine-δ-aminotransferase in the provision of glutamate for glutamine biosynthesis during pine germination. — Plant Physiol. 148: 77–88, 2008.PubMedCrossRefGoogle Scholar
  7. Chen, C.T., Chen, L.M., Lin, C.C., Kao, C.H.: Regulation of proline accumulation in detached rice leaves exposed to excess copper. — Plant Sci. 160: 283–290, 2001.PubMedCrossRefGoogle Scholar
  8. Chen, C.T., Chen, T.H., Lo, K.F., Chiu, C.Y.: Effects of proline on copper transport in rice seedlings under excess copper stress. — Plant Sci. 166: 103–111, 2004.CrossRefGoogle Scholar
  9. Chen, C.T., Kao, C.H.: Osmotic stress and water stress have opposite effects of putrescine and proline production in excised rice leaves. — Plant Growth Regul. 13: 197–202, 1993.CrossRefGoogle Scholar
  10. Chou, I.T., Chen, C.T., Kao, C.H.: Characteristics of the induction of the accumulation of proline by abscisic acid and isobutyric acid in detached rice leaves. — Plant Cell Physiol. 32: 269–272, 1991.Google Scholar
  11. Delauney, A.J., Verma, D.P.S.: Proline biosynthesis and osmoregulation in plants. — Plant J. 4: 215–223, 1993.CrossRefGoogle Scholar
  12. Feinberg, A.P., Vogelstein, B.: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. — Anal. Biochem. 132: 6–13, 1983.PubMedCrossRefGoogle Scholar
  13. Fujita, T., Maggio, A., Garcia-Rios, M., Bressan, R.A., Csonka, L.N.: Comparative analysis of regulation of expression and structures of two evolutionarily divergent genes for δ1-pyrroline-5-carboxylate synthetase from tomato. — Plant Physiol. 118: 661–674, 1998.PubMedCrossRefGoogle Scholar
  14. Funck, D., Stadelhofer, B., Koch, W.: Ornithine-δ-aminotransferase is essential for arginine catabolism but not for proline biosynthesis. — BMC Plant Biol. 8: 40, 2008.PubMedCrossRefGoogle Scholar
  15. Girousse, C., Bournocille, R., Bonnemain, J.L.: Water deficitinduced changes in concentrations in proline and some other amino acids in the phloem sap of alfalfa. — Plant Physiol. 111: 109–113, 1996.PubMedGoogle Scholar
  16. Haag-Kerwer, A., Schafer, H.J., Heiss, S., Walter, C., Rausch, T.: Cadmium exposure in Brassica juncea causes a decline in transpiration rate and leaf expansion without effect on photosynthesis. — J. exp. Bot. 341: 1827–1835, 1999.CrossRefGoogle Scholar
  17. Hervieu, F., Le Dily, F., Huault, C., Billard, J.P.: Contribution of ornithine aminotransferase to proline accumulation in NaCl-treated radish cotyledons. — Plant Cell Environ. 18: 205–210, 1995.CrossRefGoogle Scholar
  18. Hsu, Y.T., Kao, C.H.: Role of abscisic acid in cadmium tolerance of rice (Oryza sativa L.) seedlings. — Plant Cell Environ. 26: 867–874, 2003.PubMedCrossRefGoogle Scholar
  19. Hu, C.A.A., Delauney, A.J., Verma, D.P.S.: A bifunctional enzyme (Δ1-pyrroline-5-carboxylate synthetase) catalyzes the first two steps in proline biosynthesis in plants. — Proc. nat. Acad. Sci. USA 89: 9354–9358, 1992.PubMedCrossRefGoogle Scholar
  20. Jan, F.J., Pang, S.Z., Fagoaga, F., Gonsalves, D.: Turnip mosaic potyvirus resistance in Nicotiana benthamiana derived by post-transcriptional gene silencing — Transgen. Res. 8: 203–213, 1999.CrossRefGoogle Scholar
  21. Jan, F.J., Pang, S.Z., Tricoli, D.M., Gonsalves, D.: Evidences that plant developmental stage and combining transgene from different lines enhance resistance in squash mosaic comovirus coat protein transgenic plants — J. gen. Virol. 81: 2299–2306, 2000.PubMedGoogle Scholar
  22. Kandpal, R.P., Rao, N.A.: Water stress induced alterations in the properties of ornithine aminotransferase from ragi (Eleusine coracana) leaf enzymes — Biochem. Internat. 5: 297–302, 1982.Google Scholar
  23. Kastori, R., Petrovic, M., Petrovic, N.: Effects of excess lead, cadmium, copper, and zinc on water relations in sunflower — J. Plant Nutr. 15: 2427–2439, 1992.CrossRefGoogle Scholar
  24. Kiyosue, T., Yoshiba, Y., Yamaguchi-Shinozaki, K., Shinozaki, K.: A nuclear gene encoding mitochondrial proline dehydrogenase, an enzyme involved in proline metabolism, is upregulated by proline but downregulated by dehydration in Arabidopsis — Plant Cell 8: 1323–1335, 1996.PubMedCrossRefGoogle Scholar
  25. Mehta, S.K., Gaur, J.P.: Heavy-metal-induced proline accumulation and its role in ameliorating metal toxicity in Chlorella vulgaris — New Phytol. 143: 253–259, 1999.CrossRefGoogle Scholar
  26. Nagoor, S.A., Vyas, A.V.: Physiological and biochemical responses of cereal seedlings to graded levels of heavy metals. III. Effects of copper on protein metabolism in wheat seedlings — J. environ. Biol. 20: 125–129, 1999.Google Scholar
  27. Napoli, C., Lemieux, C., Jorgensen, R.: Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. — Plant Cell 2: 279–289, 1990.PubMedCrossRefGoogle Scholar
  28. Oncel, I., Keles, Y., Ustum, A.S.: Interactive effects of temperature and heavy metals stress on the growth and some biochemical compounds in wheat seedlings — Environ. Pollut. 107: 315–320, 2000.PubMedCrossRefGoogle Scholar
  29. Pang, S.Z., Jan, F.J., Gonsalves, D.: Non-target DNA sequences reduce the transgene length necessary for RNA-mediated topovirus resistance in transgenic plants — Proc. nat. Acad. Sci USA 94: 8261–8266, 1997.PubMedCrossRefGoogle Scholar
  30. Peng, Z., Lu, Q., Verma, D.P.S.: Reciprocal regulation of Δ1-pyrroline-5-carboxylate synthetase and proline dehydrogenase genes controls proline levels during and after osmotic stress in plants — Mol. gen. Genet. 253: 334–341, 1996.PubMedGoogle Scholar
  31. Pesic, P., Reggiani, R.: The process of abscisic acid-induced proline accumulation and the levels of polyamines and quaternary ammonium compounds in hydrated barley leaves — Physiol. Plant. 84: 134–139, 1992.CrossRefGoogle Scholar
  32. Roosens, N.H., Thu, T.T., Iskandar, H.M., Jacobs, M.: Isolation of the ornithine-δ-aminotransferase cDNA and effect of salt stress on its expression in Arabidopsis thaliana — Plant Physiol. 117: 263–271, 1998.PubMedCrossRefGoogle Scholar
  33. Sambrook, J., Russell, D (ed.): Molecular Cloning: a Laboratory Manual. — Cold Spring Harbor Laboratory Press, Cold Spring Harbor — New York 2001.Google Scholar
  34. Saradhi, A., Saradhi, P.P.: Proline accumulation under metal stress — J. Plant Physiol. 138: 554–558, 1991.CrossRefGoogle Scholar
  35. Savoure, A., Hua, X.J., Bertauche, N., Montagu, M., Verbruggen, N.: Abscisic acid-independent and abscisic acid-dependent regulation of proline biosynthesis following cold and osmotic stresses in Arabidopsis thaliana — Mol. gen. Genet. 254: 104–109, 1997.PubMedCrossRefGoogle Scholar
  36. Savoure, A., Jaoua, S., Hua, X.J., Ardiles, W., Van Montagu, M., Verbruggen, N.: Isolation, characterization, and chromosomal location of a gene encoding the Δ1-pyrroline -5-carboxylate synthetase in Arabidopsis thaliana — Feder. Eur. Biochem. Soc. Lett. 372: 13–19, 1995.CrossRefGoogle Scholar
  37. Schat, H., Sharma, S.S., Vooijs, R.: Heavy metal-induced accumulation of free proline in a metal-tolerant and a nontolerant ecotype of Silene vulgaris — Physiol. Plant. 101: 477–482, 1997.CrossRefGoogle Scholar
  38. Sharma, S.S., Dietz, K.J.: The significance of amino acids and amino acid-derived molecules in plant responses and adaptation to heavy metal stress — J. exp. Bot. 57: 711–726, 2006.PubMedCrossRefGoogle Scholar
  39. Siripornadulsil, S., Traina, S., Verma, D.P.S., Sayre, R.T.: Molecular mechanisms of proline-mediated tolerance to toxic heavy metals in transgenic microalgae — Plant Cell 14: 2837–2847, 2002.PubMedCrossRefGoogle Scholar
  40. Talanova, V.V., Totov, A.F., Boeva, N.P.: Effect of increasing concentrations of lead and cadmium on cucumber seedlings — Biol. Plant. 43: 441–444, 2000.CrossRefGoogle Scholar
  41. Thippeswamy, M., Chandraobulreddy, P., Sinilal, B., Shiva Kumar, M., Chinta Sudhakar: Proline accumulation and the expression of Δ1-pyrroline-5-carboxylate synthetase in two safflower cultivars — Biol. Plant. 54: 386–390, 2010.CrossRefGoogle Scholar
  42. Thomas, J.C., Malick, F.K., Endreszl, C., Davies, E.C., Murray, K.S.: Distinct responses to copper stress in the halophyte Mesembryanthemum crystallinum — Physiol. Plant. 102: 360–368, 1998.CrossRefGoogle Scholar
  43. Tripathi, A.K., Tripathi, S.: Changes in some physiological and biochemical characters in Albizia lebbek as bio-indicators of heavy metal toxicity — J. environ. Biol. 20: 93–98, 1999.Google Scholar
  44. Trotel-Aziz, P., Niogret, M.F., Larher, F.: Proline level is partly under the control of abscisic acid in canola leaf discs during recovery from hyper-osmotic stress — Physiol. Plant. 110: 376–383, 2000.CrossRefGoogle Scholar
  45. Turchetto-Zolet, A.C., Margis-Pinheiro, M., Margis, R.: The evolution of pyrroline-5-carboxylate synthase in plants: a key enzyme in proline synthesis — Mol. gen. Genet. 281: 87–97, 2009.Google Scholar
  46. Verbruggen, N., Hua, X., May, M., Montagu, M.V.: Environmental and developmental signals modulated proline homeostasis: evidence for a negative transcriptional regulator — Proc. nat. Acad. Sci. USA 93: 8787–8791, 1996.PubMedCrossRefGoogle Scholar
  47. Voetberg, G.S., Sharp, R.E.: Growth of the maize primary root at low water potentials. III. Role of increased proline deposition in osmotic adjustment. — Plant Physiol. 96: 1125–1130, 1991.PubMedCrossRefGoogle Scholar
  48. Walker, D.J., Romero, P., Correal, E.: Cold tolerance, water relations and accumulation of osmolytes in Bituminaria bituminosa. — Biol. Plant. 54: 293–298, 2010.CrossRefGoogle Scholar
  49. Wu, L., Fan, Z., Guo, L., Li, Y., Chen, Z.L., Qu, L.J.: Overexpression of the bacterial nhaA gene in rice enhances salt and drought tolerance. — Plant Sci. 168: 297–302, 2005.CrossRefGoogle Scholar
  50. Yoshiba, Y., Kiyosue, T., Nakashima, K., Yamaguchi-Shinozaki, K., Shinozaki, K.: Regulation of levels of proline as an osmolyte in plants under water stress. — Plant Cell Physiol. 38: 1095–1102, 1997.PubMedGoogle Scholar

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© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  1. 1.Department of AgronomyNational Chung Hsing UniversityTaichungTaiwan
  2. 2.Department of Plant PathologyNational Chung Hsing UniversityTaichungTaiwan
  3. 3.Department of Agriculture ChemistryNational Taiwan UniversityTaipeiTaiwan
  4. 4.Biodiversity Research CenterAcademia SinicaTaipeiTaiwan

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