Abstract
δ-OAT, ornithine-δ-aminotransferase, is the key enzyme involved in proline biosynthesis. In this study the Arabidopsis δ-OAT gene was transferred into rice (Oryza sativa L. ssp japonica cv. Zhongzuo 321), whose successful integration was demonstrated by PCR and Southern blot analysis. The over-expression of the gene in transgenic rice was also confirmed. Biochemical analysis showed that, under salt or drought stress conditions, proline contents in the leaves and roots in transgenic rice plants were 5- to 15-fold of those in non-transgenic controls. Under stress conditions, germinating rate of transgenic lines is higher than that of controls. Although the growth of rice plants tested were more and more retarded with the increasing of NaCl concentration, the transgenic plants grow faster compared to the controls under the same stress condition. Meanwhile, the resistance to KC1 and MgSO4 stresses was also found enhanced in transgenic rice. Furthermore, the over-expression of δ-OAT also improved the yield of transgenic plants under stress conditions. The average yield per plant of transgenic lines increases about 12%–41% more than that of control lines under 0.1 mol/L NaCl stress. These data indicated that the over-expression of δ-OAT, with the accumulation of proline, resulted in the enhancement of salt and drought tolerance and an increase of rice yield, which is of significance in agriculture.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Serrano, R., Macia, F. C., Moreno, V., Genetic engineering of salt and drought tolerance with yeast regulatory genes, Scientia Horticulturae, 1999, 78: 261–269.
Yoshiba, Y., Kiyosue, T., Nakashima, K. et al., Regulation of levels of proline as an osmolyte in plants under water stress, Plant Cell Physiol., 1997, 38: 1095–1102.
Hoekstra, F. A., Golovina, E. A., Buitink, J., Mechanisms of plant desiccation tolerance. Trends Plant Sci, 2001, 6: 431–438.
Delauney, A. J., Hu, C., Kishor, K. et al., Cloning of ornithine-aminotransferase cDNA by transcomplementation in Escherichia coli and regulation of proline biosynthesis, J. Biol. Chem., 1993, 268: 18673–18678.
Schobert, B., Is there an osmotic regulatory mechanism in algae and higher plants? J. Theor. Biol., 1977, 68: 17–26.
Csonka, L. N., Hanson, A. D., Prokaryotic osmoregulation: genetics and physiology, Annu. Rev. Microbiol, 1991, 45: 569–606.
Kishor, P. B. K., Hong, Z., Miao, G. H. et al., Overexpression of l-pyrroline-5-carboxylate synthetase increase proline production and confers osmotolerance in transgenic plants, Plant Physiol., 1995, 108: 1387–1394.
Solomon, A., Beer, S., Waisel, Y. et al., Effect of NaCl on the carboxylating activity of Rubisco from Tamarix jordanis in the presence and absence of proline-related compatible solutes, Physiol. Plant, 1994, 90: 198–204.
Smironff, N., Cumbes, Q. J., Hydroxyl radical scavenging activity of compatible solutes, Phytochem., 1989, 28: 1057–1060.
Saradhi, A., Saradhi, P. P., Proline accumulation under heavy metal stress, J. Plant Physiol., 1991, 138: 554–558.
Stewart, C. R., Hanson, A. D., Proline accumulation as a metabolic response to water stress, Adaptation of Plants to Water and High Temperature Stress (eds. Turner, N. C., Kramer, P. J.), New York: John Wiley & Sons, 1980, 173–189.
Venekamp, J. H., Lampe, J. E. M., Koot, J. T. M., Organic acids as sources of drought-induced proline synthesis in field bean plants, Vicia faba L, J. Plant Physiol., 1989, 133: 654–659.
Roosens, N. H., Thu, T. T., Iskandar, H. M. et al., Isolation of the ornithine-aminotransferase cDNA and effect of salt stress on its expression in Arabidopsis thaliana, Plant Physiol., 1998, 117: 263–271.
Savouré, A., Jaoua, S., Hua, X. J. et al., Isolation, characterization, and chromosomal location of a gene encoding the l-pyrroline-5-carboxylate synthetase in Arabidopsis thaliana, FEBS Letters, 1995, 372: 13–19.
Roosens, N. H., Bitar, F. A., Loenders, K. et al., Overexpression of ornithine-aminotransferase increases proline biosynthesis and confers osmotolerance in transgenic plants, Molecular Breeding, 2002, 9: 73–80.
Hong, Z., Lakkineni, K., Zhang, Z. et al., Removal of feedback inhibition of l-pyrroline-5-carboxylate synthetase results in increased proline accumulation and protection of plants from osmotic stress, Plant Physiol., 2000, 122: 1129–1136.
Zhu, B., Su, J., Chang, M. et al., Overexpression of a l-pyrroline-5-carboxylate synthetase gene and analysis of tolerance to water- and salt-stress in transgenic rice, Plant Sci., 1998, 139: 41–48.
Hervieu, F., Le Dily, L., Huault, C. et al., Contribution of ornithine aminotransferase to proline accumulation in NaCl-treated radish cotyledons, Plant Cell Environ., 1995, 18: 205–210.
Charest, C., Phan, C. T., Cold acclimation of wheat (Triticum aestivum): Properties of enzymes involved in proline metabolism, Physiol. Plant, 1990, 80: 159–168.
Hervieu, F., Le Dily, F., Billard, J. P. et al., Effects of water-stress on proline content and ornithine aminotransferase activity of radish cotyledons, Phytochemistry, 1994, 37: 1227–1231.
Yang, C. W., Wang, J. W., Kao, C. H., The relation between accumulation of abscisic acid and proline in detached rice leaves, Biol. Plant, 2000, 43: 301–314.
Roosens, N. H., Willem, R., Li, Y. et al., Proline metabolism in the wild-type and salt tolerant mutant of Nicotiana plumbaginifolia studied by 13C nuclear magnetic resonance imaging, Plant Physiol., 1999, 121: 1281–1290.
Murashige, T., Skoog, F., A revised medium for rapid growth and bioassays with tobacco tissue culture, Physiol. Plant, 1962, 15: 473–479.
Chu, C. C., Wang, C. C., Sun, C. S. et al., Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources, Scientia Sinica, 1975, 18: 659–668.
Gamborg, O. L., Miller, R. A., Ojima, K., Nutrient requirement suspension cultures of soybean root cells, Exp. Cell Res., 1968, 50: 151–158.
Murry, M. G., Thompson, W. F., Rapid isolation of high-molecularweight plant DNA, Nucl. Acids Res., 1980, 8: 8321–8325.
Clark, M. S. (ed.), Qu, L. J., Gu, H. Y. (Interps.), Chen, Z. L. (Press-corrector), Plant Molecular Biology—A Laboratory Manual, Higher Education Press Beijing and Springer-Verlag Berlin Heidelberg, 1998, 8.
Bates, L. S., Rapid determination of free proline for water stress studies, Plant Soil, 1973, 39: 205–207.
Chen, Y., Zhao, M., Li, Z. P. et al., The function of the nuclear matrix attachment region of silkworm rDNA as an autonomously replicating sequence in plasmid and chromosomal replication origin in yeast, Biochem. Biophys. Res. Commun., 2002, 299: 723–729.
Cheng, Z., Targolli, J., Wu, R., Tobacco matrix attachment region sequence increased transgene expression levels in rice plants, Molecular Breeding, 2001, 7: 317–327.
Bengston, C., Klockare, B., Klockare, R. et al., The after-effect of water stress on chlorophyll formation during greening and the levels of abscisic acid and proline in dark grown wheat seedlings, Physiol. Plant, 1978, 43: 205–212.
Chen, C. T., Chen, L. M., Lin, C. C. et al., Regulation of proline accumulation in detached rice leaves exposed to excess copper, Plant Sci., 2001, 160: 283–290.
Serrano, R., Gaxiola, R., Microbial models and salt stress tolerance in plants, Cri. Rev. Plant Sci., 1994, 13: 121–138.
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
Wu, L., Fan, Z., Guo, L. et al. Over-expression of an Arabidopsis δ-OAT gene enhances salt and drought tolerance in transgenic rice. Chin.Sci.Bull. 48, 2594–2600 (2003). https://doi.org/10.1360/03wc0218
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1360/03wc0218