Scientia Marina, Vol 80, No 1 (2016)

Identification of Chattonella (Raphidophyceae) species in long-term phytoplankton samples from Santa Giusta Lagoon, Italy

Daniela Stacca
Dipartimento di Architettura, Design e Urbanistica, University of Sassari, Italy

Cecilia Teodora Satta
Dipartimento di Architettura, Design e Urbanistica, University of Sassari, Italy

Silvia Casabianca
Dipartimento di Scienze Biomolecolari, University of Urbino, Italy

Antonella Penna
Dipartimento di Scienze Biomolecolari, University of Urbino, Italy

Bachisio Mario Padedda
Dipartimento di Architettura, Design e Urbanistica, University of Sassari, Italy

Nicola Sechi
Dipartimento di Architettura, Design e Urbanistica, University of Sassari, Italy

Antonella Lugliè
Dipartimento di Architettura, Design e Urbanistica, University of Sassari, Italy


Chattonella species in a Mediterranean lagoon (Santa Giusta Lagoon, Sardinia, Italy) were identified by applying a molecular approach to fixed natural phytoplankton samples collected over the last two decades. Like the other raphidophytes, Chattonella cells are naked and lose their shape when fixed, making species identification difficult on the basis of their morphological characteristics. Employing species-specific primers (oBTG-005-F, oBTG-027-R, oBTG-028-R) for the amplification of the ITS-5.8S rDNA region, we established the occurrence of C. subsalsa in fixed natural phytoplankton samples collected in coincidence with fish death events. Additionally, we established the presence of the recently discovered C. cf. subsalsa Adriatic genotype by analysing cellular cultures obtained from the same lagoon in 2013. This is the second worldwide record of C. cf. subsalsa Adriatic genotype. Our results revealed that the species-specific primers oBTG-005-F and oBTG-028-R distinguished this new genotype only when present singularly. This study provides valuable data that increase knowledge of C. subsalsa genotypes and of the long-term occurrence of Chattonella blooms in a transitional ecosystem through the use of samples up to 20 years old.


Chattonella subsalsa genotypes; transitional ecosystems; harmful algal blooms; LTER-Italy; ITS-5.8S rDNA; LSU rDNA

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Adachi M., Sako Y., Ishida Y. 1994. Restriction fragment length polymorphism of ribosomal DNA internal transcribed spacer and 5.8S regions in Japanese Alexandrium species (Dinophyceae). J. Phycol. 30: 857-863.

Attaran-Fariman G., Bolch C.J.S. 2014. Morphology and genetic affinities of a novel Chattonella isolate (Raphidophyceae) isolated from Iran's south coast (Oman Sea). Turk. J. Bot. 38: 156-168.

Band-Schmidt C.J., Morquecho L., Hernandez-Becerril D.U., et al. 2004. Raphidophyceans on the coasts of Mexico. Hydrobiologia 515: 79-89.

Band-Schmidt C.J., Martínez-López A., Bustillos-Guzmán J.J., et al. 2012. Morphology, biochemistry, and growth of raphidophyte strains from the Gulf of California. Hydrobiologia 693: 81-97.

Ben Ali A., De Baere R., De Wachter R., et al. 2002. Evolutionary relationships among heterokont algae (the autotrophic stramenopiles) based on combined analyses of small and large subunit ribosomal RNA. Protist 153(2): 123-132. PMid:12125754

Biecheler B. 1936. Sur une Chloromonadine nouvelle d'eau saumatre Chattonella subsalsa n. gen., n. sp. Arch. Zool. Exp. Gen. 78: 79-83.

Bowers H., Tomas C., Tengs T., et al. 2006. Raphidophyceae [Chadefaud ex Silva] systematics and rapid identification: sequence analyses and real-time PCR assays. J. Phycol. 42: 1333-1348. PMid:20411032 PMCid:PMC2856949

Connell L.B. 2000. Nuclear ITS region of the alga Heterosigma akashiwo (Chromophyta: Raphidophyceae) is identical in isolates from Atlantic and Pacific basins. Mar. Biol. 136: 953-960.

Connell L. 2002. Rapid identification of marine algae (Raphidophyceae) using three-primer PCR amplification of nuclear internal transcribed spacer (ITS) regions from fresh and archived material. Phycologia 41(1): 15-21.

Demura M., Noël M.H., Kasai F., et al. 2009. Taxonomic revision of Chattonella antiqua, C. marina and C. ovata (Raphidophyceae) based on their morphological characteristics and genetic diversity. Phycologia 48(6): 518-535.

Guillard R.R.L., Hargraves P.E. 1993. Stichochrysis immobilis is a diatom, not a chrysophyte. Phycologia 32(3): 234-236.

Hall T.A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl. Acid. S. 41: 95-98.

Hallegraeff G.M., Hara Y. 2003. Taxonomy of harmful marine raphidophytes. In: Hallegraeff G.M., Anderson D.M., Cembella A.D. (eds), Manual on harmful marine microalgae. Monographs on Oceanographic Methodology. Methodology 11, Unesco Paris, pp. 511-522.

Hallegraeff G.M., Munday B., Baden D., et al. 1998. Chattonella marina (Raphidophyte) bloom associated with mortality of cultured bluefin tuna (Thunnus maccoyii) in south Australia. In: Reguera B., Blanco J., Fernandez M.L., et al. (eds), Harmful Algae. Xunta de Galicia and IOC-UNESCO, Vigo, Paris, pp. 93-96.

Imai I., Yamaguchi M., Hori Y. 2006. Eutrophication and occurrences of harmful algal blooms in the Seto Inland Sea. Plank. Benth. Res. 1: 71-84.

Imai I., Yamaguchi M. 2012. Life cycle, physiology, ecology and red tide occurrences of the fish-killing raphidophyte Chattonella. Harmful Algae 14: 46-70.

Jugnu R., Kripa V. 2009. Effect of Chattonella marina [(Subrahmanyan) Hara et Chihara 1982] bloom on the coastal fishery resources along Kerala coast, India. Indian J. Mar. Sci. 38: 77-88.

Katoh K., Misawa K., Kuma K., et al. 2002. MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res. 30(14): 3059-3066. PMid:12136088 PMCid:PMC135756

Kim S.Y., Seo K.S., Lee C.K., et al. 2007. Diurnal modification of a red tide causing organisms, Chattonella antiqua (Raphidophyceae) from Korea. Algae 22: 95-106.

Klöpper S., John U., Zingone A., et al. 2013. Phylogeny and morphology of a Chattonella (Raphidophyceae) species from the Mediterranean Sea: what is C. subsalsa? Eur. J. Phycol. 48(1):79-92.

Lewitus A.J., Brock L.M., Burke M.K., et al. 2008. Lagoonal stormwater detention ponds as promoters of harmful algal blooms and eutrophication along the South Carolina coast. Harmful Algae 8: 60-65.

Lugliè A., Sechi N., Oggiano G., et al. 2002. Ecological assessment of Santa Giusta Lagoon (Sardinia, Italy). Ann. Chim-Rome 92: 239-247.

Marin III R., Scholin C. 2010. Sandwich Hybridization. In: Karlson B., Cusack C., Bresnan E. (eds), Microscopic and molecular methods for quantitative phytoplankton analysis. IOC Man Guides 55, Unesco Paris, pp 87-110.

Mikhail S.K. 2007. First monospecific bloom of the harmful raphidophyte Chattonella antiqua (Hada) Ono in Alexandria waters related to water quality and copepod grazing. Chem. Ecol. 23(5): 393-407.

Penna A., Bertozzini E., Battocchi C., et al. 2007. Monitoring of HAB species in the Mediterranean Sea through molecular techniques. J. Plankton Res. 29: 19-38.

Satta C.T., Anglès S., Garcès E., et al. 2014. Dinoflagellate cyst assemblages in surface sediments from three shallow Mediterranean Lagoons (Sardinia, North Western Mediterranean Sea). Estuar. Coasts 37: 646-663.

Scholin C.A., Herzog M., Sogin M., et al. 1994. Identification of group and strain-specific genetic markers for globally distributed Alexandrium (Dinophyceae). 2. Sequence analysis of a fragment of the LSU rRNA gene. J. Phycol. 30(6): 999-1011.

Sechi N., Fiocca F., Sannio A., et al. 2001. Santa Giusta Lagoon (Sardinia): phytoplankton and nutrients before and after waste water diversion. J. Limnol. 60(2): 194-200.

Stamatakis A. 2006. RAxML-VI-HPC: Maximum Likelihood-based Phylogenetic Analyses with Thousands of Taxa and Mixed Models. Bioinformatics 22(21): 2688-2690. PMid:16928733

Subrahmanyan R. 1954. On the life-history and ecology of Hornellia marina gen. et sp. nov., (Chloromonadineae), causing green discoloration of the sea and mortality among marine organisms off the Malabar Coast. Indian J. Fish. 1: 182-203.

Tomas C. 1998. Blooms of potentially harmful raphidophycean flagellates in Florida coastal waters. In: Reguera B., Blanco J., Fernandez M.L., et al. (eds), Harmful Algae. Xunta de Galicia and IOC-UNESCO, pp. 101-103.

Tseng C.K., Zhou M.J., Zou J.Z. 1993. Toxic phytoplankton studies in China. In: Smayda T.J., Shimizu Y. (eds), Toxic Phytoplankton Blooms in the Sea. Elsevier, New York, pp. 347-352.

Utermöhl H. 1958. Zur vervollkommung der quantitativen phytoplankton-methodik. Verkandlungen der Internationalen Vereinigung für Theoretische und Angewandte, Limnol. 9: 1-39.

Zhang Y., Fu F.-X., Whereat E., Coyne K.J., Hutchins D.A. 2006. Bottom-up controls on a mixed-species HAB assemblage: A comparison of sympatric Chattonella subsalsa and Heterosigma akashiwo (Raphidophyceae) isolates from the Delaware Inland Bays. Harmful Algae 5: 310-320.

Zingone A., Siano R., D'Alelio D., et al. 2006. Potentially toxic and harmful microalgae from coastal waters of the Campania region (Tyrrhenian Sea, Mediterranean Sea). Harmful Algae 5: 321-337.

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