Abstract
The unicellular green alga Dunaliella is outstanding for its ability of massive accumulation of carotenoids. To elucidate the carotenoids synthesis pathway in this alga, phytoene desaturase (pds) gene cDNA together with its DNA sequences were isolated and their structures and functions analyzed. The full-length pds cDNA of 2290 bp (GenBank Accession No. DQ243892) was deduced from RACE results, including untranslated 21 bp 5′-and 520 bp 3′-flanking regions and an open reading frame of 582 amino acids, coding a protein of 64.196 kDa. The DNA sequence of 2908 bp (GenBank Accession No. DQ845248) including five introns was obtained. The fifth intron was uncompleted and complex, including two bases’ perfect repeats (GT)10 and large different-sized repeats within the last 400 bp. The Southern blot hybridization result demonstrated that this gene occurred as a single copy in this species, and the quantitative RT-PCR result showed that the transcription of this gene was constitutive. The evolutional significance of pds was discussed.
Similar content being viewed by others
References
Barkovich, R., and J. C. Liao, 2001. Metabolic engineering of isoprenoids. Metab. Eng., 3(1): 27–39.
Bartley, G. E, P. V. Viitanen, and I. Pecker, 1991. Molecular cloning and expression in photosynthetic bacteria of a soybean cDNA coding for phytoene desaturase, an enzyme of the carotenoid biosynthesis pathway. Proc. Natl. Acad. Sci., 88(15): 6532–6536.
Bartley, G. E, P. A. Scolnik, and P. Beyer, 1999. Two Arabidopsis thaliana carotene desaturases, phytoene desaturase and z-carotene desaturase, expressed in Escherichia coli, catalyze a poly-cis pathway to yield pro-lycopene. Eur. J. Biochem., 259(3): 396–403.
Ben-Amotz, A, A. Katz, and M. Avron, 1982. Accumulation of β-carotene in halotolerant algae: purification and charcaterization of β-carotene rich globules from Dunaliella bardwil. J. Phycol., 18(4): 529–537.
Borowitzka, M. A., and L. J. Borowitzka, 1988. Micro-algal Biotechnogy. Cambridge University Press, Cambridge, 477pp.
Brown, A. D., and L. J. Borowitzka, 1979. Halotolerance of Dunaliella. In: Biochemistry and Physiology of Protozoa. 2nd edition. Levandowsky, M., and Hutner, S. H., eds., Academic Press, New York, 1: 139–190.
Goodwin, T. W., 1980. The Biochemistry of the Carotenoids. 2nd edition. Vol.1. ISBN: 0412216906 0412237709, Chapman and Hall, London, 21–130.
Cunningham, F. X., and E. Jr, Gantt, 1998. Genes and enzymes of carotenoid biosynthesis in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol., 49(1): 557–583.
Jiang, G. Z., P. R. Xu, and X. L. Niu, 2005. Cloning of cytosolic hsp70 cDNA in Dunaliella salina and the expression of hsp70 Mrna. Mar. Sci., 29(5): 43–49.
Jiang, J. G., Y. Yan, and D. L. Song, 2004. Cloning and Homology Analysis of the Conservative Sequence of Phytoene Synthase Gene from Dunaliella Salina. J. South Chin. Univ. Tech. (Natural Science), 32(10): 76–79.
Johnson, E. A., and W. A. Schroeder, 1996. Microbial carotenoids. Adv. Biochem. Eng./Biotech., 53: 119–178.
Jukes, T. H., 1996. On the prevalence of certain codons (’RNY’) in genes for proteins. J. Mol. Evol., 42(4): 377–381.
Kato, M., Y. Ikoma, and H. Matsumoto, 2004. Accumulation of carotenoids and expression of carotenoid biosynthetic genes during maturation in citrus fruit. Plant Physiol., 134(2): 824–837.
Knekt, P., R. Jaervinen, R. Seppaenen, A. Rissanen, P. Knekt, R. Jarvinen, et al., 1991. Dietary antioxidants and the risk of lung cancer. Am. J. Epidemiol. 134: 471–479.
Kück, U., Y. Choquet, and M. Schneider, 1987. Structural and transcription analysis of two homologous genes for the P700 chlorophyll a-apoproteins in Chlamydomonas reinhardii: evidence for in vivo trans-splicing. The EMBO J., 6(8): 2185–2195.
Linden, H., A. Vioque, and G. Sandmann, 1993. Isolation of a carotenoid biosynthesis gene coding for ζ-carotene desaturase from Anabaena PCC 7120 by heterologous complementation. FEMS Microbiol Lett., 106: 99–104.
Liu, J. G., C. Y. Wu, and H. H. Liu, 1993. An optimized model for the culture of Dunaliella salina. In: Proceedings of the Second International Conference on the Marine Biology of the South China Sea., World Publishing Corporation, Beijing, 69.
Loeblich, L. A., 1982. Photosynthesis and pigments influenced by light intensity and salinity in the halophile Dunaliella salina (Chlorophyta). J. Mar. Biol., Assoc. UK, 62(8): 493–508.
Metting, F. B., 1996. Biodiversity and application of microalgae. J. Indus. Microbiol. Biotech., 17(5–6): 477–489.
Pecker, I., D. Chamovitz, and H Linden, 1992. A single polypeptide catalyzing the conversion of phytoene to I-carotene is transcriptionally regulated during tomato fruit ripening. Proc. Natl. Acad. Sci., 89(11): 4962–4966.
Rabbani, S., P. Beyer, and J. Vonlintig, 1998. Induced β-carotene synthesis driven by triacylglycerol deposition in the unicellular alga Dunaliella bardawil. Plant Physiol., 116(4): 1239–1248.
Ramakrishna, W., A. P. Davierwala, and V. S. Gupta, 1998. Expansion of a (GA) di-nucleotide at a microsatellite locus associated with domestication in rice. Biochem. Gen., 36(9–10): 323–327.
Rock, C. D., and J. A. D. Zeevaart, 1991. The aba mutant of Arabidopsis thaliana is impaired in poxy-carotenoid biosynthesis. Proc. Natl. Acad. Sci., 88(17): 7496–7499.
Sandmann, G., 2001. Carotenoid biosynthesis and biotechnological application. Archives of biochemistry and biophysics, 385(1): 4–12.
Shepherd, J. C., 1981. Method to determine the reading frame of a protein from the urine/pyrimidine genome sequence and itspossible evolutionary justification. Proc. Natl. Acad. Sci., 78(3): 1596–1600.
Silflow, C. D., R. L. Chisholm, and T. W. Conner, 1985, The two alpha-tubulin genes of Chlamydomonas reinhardi code for slightly different proteins. Mol. Cell. Biol., 5(9): 2389–2398.
Stahelin, H. B., K. F. Gey, and M. Eichholzer, 1991, Plasma antioxidant vitamins and subsequent cancer mortality in the 12-year follow-up of the prospective Basel Study. American J. Epidemiol., 133(8): 766–775.
Wu, K. S., and S. D. Tanksley, 1993. Abundance, polymorphism and genetic mapping of microsatellites in rice. Mol. Gen. Genet., 241(1–2): 225–235.
Vander Ploeg, L. H., A. Y. Liu, and P. A. Michels, 1982. RNA splicing is required to make the messenger RNA for a variant surface antigen in trypanosomes. Nucleic Acids Res., 10(12): 3591–3604.
Yang, X. M., 1996. Effects of cell density and culture medium ‘ageing’ on the growth and β-carotene accumulation of halotolerent alga Dunaliella salina. Mar. Sci., 6: 39–44.
Zheng, M., L. Bai, F. Ma, and Q. He, 2003. Molecular Cloning of cbr Gene in Dunaliella Salina. J. Sichuan Univ. (Natural Science Edition), 40(3): 590–593.
Zhu, Y. H., J. G. Jiang, and Q. S. Lin, 2005. Cloning and sequence analysis of phytoene desaturase cDNA from Dunaliella salina. Food Ferment Ind., 31(9): 21–23.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sun, G., Sui, Z. & Zhang, X. Cloning and characterization of the phytoene desaturase (pds) gene — a key enzyme for carotenoids synthesis in Dunaliella (Chlorophyta). J. Ocean Univ. China 7, 311–318 (2008). https://doi.org/10.1007/s11802-008-0311-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11802-008-0311-y