Abstract
Dunaliella salina has become the most important microorganism for the production of β-carotene around the world. Natural carotenoids are a source of active metabolites utilized in different areas of food nutrition and pharmaceuticals, both in humans and also in animals. Identification of Dunaliella species from natural environments or certified culture collections is not precise and it is time consuming. However, accurate identification is extremely important because a slight difference in Dunaliella species generates great differences in carotenoids production. Here, we describe an intron-sizing method to make a rapid and precise identification for each of the most important carotenogenic species, showing that each hyperproducer species has an exclusive 18S rDNA fingerprint profile.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Oren A (2005) A hundred years of Dunaliella research: 1905–2005. Saline Syst 1:2
Borowitzka MA, Siva CJ (2007) The taxonomy of the genus Dunaliella (Chlorophyta, Dunaliellales) with emphasis on the marine and halophilic species. J Appl Phycol 19: 567–590
Ben-Amotz A, Asís A, Avron M (1991) The biotechnology of cultivating Dunaliella for production of β-carotene rich algae. Bioresour Technol 38:233–235
Gómez PI, González MA (2004) Genetic variation among sever strains of Dunaliella salina (Chlorophyta) with industrial potencial, based on RAPD bandung patterns and on nuclear ITS rDNA sequences. Aquaculture 233:149–162
Olmos SJ, Paniagua MJ, Contreras FR (2000) Molecular identification of Dunaliella sp. utilizing the 18S rDNA gene. Lett Appl Microbiol 30:80–84
García TA, Olmos SJ (2007) Quantification by fluorescent in situ hybridization of bacteria associated with Litopenaeus vannamei larvae in Mexican shrimp hatchery. Aquaculture 262:211–218
Hernández ZG, Olmos SJ (2006) Identification of bacterial diversity in the oyster Crassostrea gigas by fluorescent in situ hybridization and polymerase chain reaction. J Appl Microbiol 100:664–672
DeLong EF, Wickham GS, Pace NR (1989) Phylogenetic stains: ribosomal RNA-based probes for the identification of single cells. Science 243:1360–1363
Ki SJ, Han MS (2005) Sequence-based diagnostics and phylogenetic approach of uncultured freshwater dinoglagellate Peridinium (Dinophyceae) species, based on single-cell sequencing of rDNA. J Appl Phycol 17:147–153
Ki JS, Han MS (2007) Rapid molecular identification of the harmful freshwater dinoglagellate Peridinium in varios life stages using genus-specific single-cell PCR. J Appl Phycol 19:467–470
Olmos SJ, Paniagua MJ, Contreras FR, Trujillo L (2002) Molecular identification of β-carotene hyper-producing strain of Dunaliella from saline environments using specie-specific oligonucleotides. Biotechnol Lett 24:365–369
Olmos SJ, Ochoa SJL, Paniagua-Michel JJ, Contreras FR (2009) DNA fingerprinting differentiation between β-carotene hyperproducer strain of Dunaliella from around the world. Saline Syst 5:5
Guillard RRL, Sieracki MS (2005) Counting cells in cultures with the light microscope. In: Andersen RA (ed) Algal culturing techniques. Elsevier Academic Press, Burlington, p 239
Moore DD (1992) Reagent and solutions. Appendix 1. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) Short protocols in molecular biology. Wiley, New York, pp A1–57
Guillard RRL (2005) Purification methods for microalgae. In: Andersen RA (ed) Algal culturing techniques. Elsevier Academic Press, Burlington, p 117
Harrison PJ, Berges JA (2005) Marine culture media. In: Andersen RA (ed) Algal culturing techniques. Elsevier Academic Press, Burlington, p 21
Andersen RA, Berges JA, Harrison PJ, Watanabe MM (2005) Recipes for freshwater and seawater media. Appendix A. In: Andersen RA (ed) Algal culturing techniques. Elsevier Academic Press, Burlington, p 507
Ben-Amotz A (2004) Industrial production of microalgal cell-mass and secondary products—major industrial species Dunaliella. In: Richmond A (ed) Handbook of microalgae culture: biotechnology and applied phycology. Blackwell Science Ltd, Oxford, UK, p 273
Andersen RA, Kawachi M (2005) Traditional microalgae isolation techniques. In: Andersen RA (ed) Algal culturing techniques. Elsevier Academic Press, Burlington, p 83
Moore DD (1992) Preparation and analysis of DNA. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) Short protocols in molecular biology. Wiley, New York, pp 2–1
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, NY (vols. 1, 2, 3)
Innis MA, Gelfand DH (1990) Optimization of PCRs. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic, San Diego, p 3
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Olmos-Soto, J., Paniagua-Michel, J., Contreras, R., Ochoa, L. (2012). DNA Fingerprinting Intron-Sizing Method to Accomplish a Specific, Rapid, and Sensitive Identification of Carotenogenic Dunaliella Species. In: Barredo, JL. (eds) Microbial Carotenoids from Bacteria and Microalgae. Methods in Molecular Biology, vol 892. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-879-5_16
Download citation
DOI: https://doi.org/10.1007/978-1-61779-879-5_16
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-878-8
Online ISBN: 978-1-61779-879-5
eBook Packages: Springer Protocols