Abstract
Since the first formal description of the unicellular green algae Dunaliella salina, its presence in hypersaline environments worldwide and its physiological responses to different environmental conditions have been studied extensively. Moreover, due to massive carotenoid accumulation by some strains under specific growth conditions, its biotechnological applications have attracted a great deal of scientific interest. In this study, the phylogenetic relationship, growth, and carotenogenesis of a new strain of Dunaliella salina isolated from Maharlu salt lake in Shiraz (latitude 29.26°N, longitude 52.48°E), Iran were investigated. First, a phylogram based on neighbor-joining analysis of the nuclear rDNA ITS (ITS-1 + 5.8 rDNA + ITS-2) sequence data was constructed. The phylogenetic tree showed that the new isolate is part of a major clade containing several strains of D. salina and was designated as D. salina MSI-1. Then, the responses of the new isolate to the initial pH of the culture media and different concentrations of nitrate, NH4 +, and citrate were examined. As with other strains of D. salina, growth and carotenogenesis were controlled by the levels of nitrate and NH4 + in the growth media. Low available nitrogen negatively affected growth but enhanced carotenoid accumulation. Insensitivity of carotenogenesis to citrate indicates a minor contribution of cytosolic IPP synthesis to the overall carotenoid production. Despite changes in the initial pH of the culture media over the experimental period, the initial pH had marked effects on the growth and carotenogenesis of the new isolate. These effects, together with the higher cell carotenoid content observed at pH 11.0, await further research. The results confirm that the analysis of the ITS sequences is a reliable basis for determination of the genetic relatedness among strains of the genus Dunaliella, and the search for strains with novel characteristics may have valuable biotechnological applications.
Similar content being viewed by others
References
Abd EL-Baky, H. H., F. K. El-Baz & G. S. El-Baroty, 2004. Production of antioxidant by the green alga Dunaliella salina. International Journal of Agriculture and Biology 6: 49–57.
Ben-Amotz, A. & M. Avron, 1990. The biotechnology of cultivating the halotolerant alga Dunaliella. Trends in Biotechnology 8: 121–126.
Borowitzka, M. A. & L. J. Borowitzka, 1988a. Dunaliella. In Browitzka, M. A. & L. J. Borowitzka (eds), Micro-algal Biotechnology. Cambridge University Press, Cambridge, DC: 27–58.
Borowitzka, M. A. & L. J. Borowitzka, 1988b. Limits to growth and carotenogenesis in laboratory and large-scale outdoor cultures of D. salina. In Stadler, T., J. Mollion, M. C. Berdus, Y. Karamanos, H. Morvan & D. Christiane (eds), Alga Biotechnology. Elsevier Applied Science, Barking DC: 139–150.
Borowitzka, M. A. & C. J. Siva, 2007. The taxonomy of the genus Dunaliella (Chlorophyta, Dunaliellales) with emphasis on the marine and halophilic species. Journal of Applied Phycology 19: 567–590.
Borowitzka, L. J., M. A. Borowitzka & T. Moulton, 1984. Mass culture of Dunaliella: from laboratory to pilot plant. Hydrobiologia 116(117): 115–121.
Borowitzka, M. A., L. J. Borowitzka & D. Kessly, 1990. Effects of salinity increase on carotenoid accumulation in the green alga Dunaliella salina. Journal of Applied Phycology 2: 11–119.
Choudhari, S. & R. Singhal, 2008. Media optimization for the production of β-carotene by Blakeslea trispora: a statistical approach. Bioresource Technology 99: 722–730.
Coesel, S. N., A. C. Baumgartner, L. M. Teles, A. A. Ramos, N. M. Henriques, L. Cancela & J. C. S. Varela, 2008. Nutrient limitation is the main regulatory factor for carotenoid accumulation and for Psy and Pds steady state transcript levels in Dunaliella salina (Chlorophyta) exposed to high light and salt stress. Marine Biotechnology 10: 602–611.
Coleman, A. W. & J. C. Mai, 1997. Ribosomal DNA ITS-1 and ITS-2 sequence comparison as a tool for predicting genetic relatedness. Journal of Molecular Evolution 44: 258–271.
Eijckelhoff, C. & J. P. Dekker, 1997. A routine method to determine the chlorophyll a, pheophytin a, and β-carotene contents of isolated photosystem II reaction center complexes. Photosynthesis Research 52: 69–73.
Fazeli, M. R., H. Tofighi, N. Samadi, H. Jamalifar & A. Fazel, 2006. Carotenoid accumulation by Dunaliella tertiolecta (Lake Urmia isolate) and Dunaliella salina (CCAP 19/18 & WT) under stress conditions. Daru 14(3): 146–150.
Forage, R. G., D. E. F. Harrisen & D. E. Pitt, 1985. Effect of environment on microbial activity. In Moo-yong M., (ed.), Comprehensive Biotechnology. Pergamon Press, DC: 263–270.
Gomez, P. I. & M. A. Gonzalez, 2001. Genetic polymorphism in eight Chilean of the carotenogenic microalga Dunaliella salina Teodoresco (Chlorophyta). Biological Research 34: 23–30.
Gomez, I. P. & M. A. Gonzalez, 2004. Genetic variation among seven strains of Dunaliella salina (Chlorophyta) with industrial potential, based on RAPD banding patterns and on nuclear ITS rDNA sequences. Aquaculture 233: 149–162.
Gonzalez, M. A., P. I. Gomez & R. Montoya, 1999. Comparison of PCR-RFLP analysis of the ITS region with morphological criteria of various strains of Dunaliella. Journal of Applied Phycology 10: 573–580.
Gonzalez, M. A., A. W. Coleman, P. I. Gomez & R. Montoya, 2001. Phylogenetic relationship among various strains of Dunaliella (Chlorophyceae) based on nuclear ITS rDNA sequences. Journal of Phycology 37: 604–611.
Hai-Zhu, J., G. Cheng-Hua, H. Jun, W. Qing-Hau, J. Zhu-Mao & S. Fu-Zhang, 1998. Effects of Mg2+, NaCl, citric acid and other factors on synthesis and accumulation of β-carotene in Dunaliella salina. Chinese Journal of Oceanology and Limnology 16: 364–368.
Javor, B., 1989. Hypersaline Environments. Microbiology and Biogeochemistry. Springer-Verlag, Berlin DC: 328–335.
Jimenez, C. & F. X. Niell, 1991. Growth of Dunaliella viridis Teodoresco: effect of salinity, temperature and nitrogen concentration. Journal of Applied Phycology 3: 319–327.
Kim, S. W., W. T. Seo & Y. H. Park, 1996. Increased β-carotene synthesis in Blakeslea trisporen under strong alkaline culture condition. Biotechnology Letters 18: 1287–1290.
Lichtenthaler, H. K., M. Rohmer & J. Schwender, 1997. Two independent biochemical pathways for isopentenyl diphosphate (IPP) and isoprenoid biosynthesis in higher plants. Physiologia Plantarum 101: 643–652.
Loeblich, L. A., 1982. Photosynthetic and pigments influenced by light intensity and salinity in the halophilic Dunaliella salina (Chlorophyta). Journal of Marine Biological Association of the United Kingdom 62: 493–508.
Marin, N., F. Morales, C. Lodeiros & E. Tamigneaux, 1998. Effect of nitrate concentration on growth and pigment synthesis of Dunaliella salina cultivated under low illumination and preadapted to different salinities. Journal of Applied Phycology 10: 405–411.
Nikookar, K., A. Moradshahi & M. Kharati, 2004. Influence of salinity on the growth, pigmentation and ascorbate peroxidase activity of Dunaliella salina isolated from Maharlu salt lake in Shiraz. Iranian Journal of Science and Technology 28: 117–125.
Oren, A., 2005. A hundred years of Dunaliella research: 1905–2005. Saline systems 1: 2.
Ramos, A., S. Coesel, A. Marques, M. Rodrigues, A. Baumgartner, J. Noronha, A. Rauter, B. Brenig & J. Varela, 2008. Isolation and characterization of a stress-inducible Dunaliella salina Lcy-β gene encoding a functional lycopene β-cyclase. Applied Microbiology and Biotechnology 79: 819–828.
Ruckert, G. V. & A. Giani, 2004. Effect of nitrate and ammonium on the growth and protein concentration of Microcystis viridis Lemmermann (Cyanobacteria). Revista Brasileira de Botanica 27: 325–331.
Tamura, K., J. Dudley, M. Nei & S. Kumar, 2007. MEGA 4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology Evolution 24(8): 1596–1599.
Venkatesh, N. S., R. Balaji & P. Satnyamurthy, 2005. Medium for the production of betacarotene and other carotenoids from Dunaliella salina (ARL 5) and a strain of Dunaliella salina for production of carotenes using the novel media. U.S. Patent 6936459 B1.
Acknowledgments
The authors would like to thank the Shiraz University Research Council for supporting this research.
Author information
Authors and Affiliations
Corresponding author
Additional information
Handling editor: Luigi Naselli-Flores
Rights and permissions
About this article
Cite this article
Zamani, H., Moradshahi, A. & Karbalaei-Heidari, H.R. Characterization of a new Dunaliella salina strain MSI-1 based on nuclear rDNA ITS sequences and its physiological response to changes in composition of growth media. Hydrobiologia 658, 67–75 (2011). https://doi.org/10.1007/s10750-010-0450-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10750-010-0450-1