Biologia plantarum 2012, 56:337-343 | DOI: 10.1007/s10535-012-0095-1

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

H. -M. Ku1, C. -W. Tan1,2, Y. -S. Su3, C. -Y. Chiu4, C. -T. Chen3,*, F. -J. Jan2,*
1 Department of Agronomy, National Chung Hsing University, Taichung, Taiwan
2 Department of Plant Pathology, National Chung Hsing University, Taichung, Taiwan
3 Department of Agriculture Chemistry, National Taiwan University, Taipei, Taiwan
4 Biodiversity Research Center, Academia Sinica, Taipei, Taiwan

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.

Keywords: abscisic acid; gene expression; tobacco
Subjects: abscisic acid; gene expression; water stress; copper; proline; water content; proteins; ornithine aminotransferase; proline dehydrogenase; pyrroline-5-carboxylate synthetase
Species: Nicotiana benthamiana

Received: May 24, 2010; Accepted: February 15, 2011; Published: June 1, 2012Show citation

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Ku, H.-M., Tan, C.-W., Su, Y.-S., Chiu, C.-Y., Chen, C.-T., & Jan, F.-J. (2012). The effect of water deficit and excess copper on proline metabolism in Nicotiana benthamiana. Biologia plantarum, 56(2), 337-343. doi: 10.1007/s10535-012-0095-1.
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    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. Go to original source...
    2. Bassi, R., Sharma, S.S.: Proline accumulation in wheat seedlings exposed to zinc and copper. - Phytochemistry 33: 1339-1342, 1993. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    11. Delauney, A.J., Verma, D.P.S.: Proline biosynthesis and osmoregulation in plants. - Plant J. 4: 215-223, 1993. Go to original source...
    12. Feinberg, A.P., Vogelstein, B.: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. - Anal. Biochem. 132: 6-13, 1983. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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.
    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. Go to original source...
    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. Go to original source...
    19. Hu, C.A.A., Delauney, A.J., Verma, D.P.S.: A bifunctional enzyme (Delta;1-pyrroline-5-carboxylate synthetase) catalyzes the first two steps in proline biosynthesis in plants. - Proc. nat. Acad. Sci. USA 89: 9354-9358, 1992. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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.
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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.
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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.
    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. Go to original source...
    33. Sambrook, J., Russell, D (ed.): Molecular Cloning: a Laboratory Manual. - Cold Spring Harbor Laboratory Press, Cold Spring Harbor - New York 2001.
    34. Saradhi, A., Saradhi, P.P.: Proline accumulation under metal stress - J. Plant Physiol. 138: 554-558, 1991.
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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.
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...
    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. Go to original source...

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