Phylogenetic relationships of Pseudo-nitzschia subpacifica (Bacillariophyceae) from the Mexican Pacific, and its production of domoic acid in culture

Pseudo-nitzschia is a cosmopolitan genus, some species of which can produce domoic acid (DA), a neurotoxin responsible for the Amnesic Shellfish Poisoning (ASP). In this study, we identified P. subpacifica for the first time in Todos Santos Bay and Manzanillo Bay, in the Mexican Pacific using SEM and molecular methods. Isolates from Todos Santos Bay were cultivated under conditions of phosphate sufficiency and deficiency at 16°C and 22°C to evaluate the production of DA. This toxin was detected in the particulate (DAp) and dissolved (DAd) fractions of the cultures during the exponential and stationary phases of growth of the cultures. The highest DA concentration was detected during the exponential phase grown in cells maintained in P-deficient medium at 16°C (1.14 ± 0.08 ng mL-1 DAd and 4.71 ± 1.11 × 10−5 ng cell-1 of DAp). In P-sufficient cultures DA was higher in cells maintained at 16°C (0.25 ± 0.05 ng mL-1 DAd and 9.41 ± 1.23 × 10−7 ng cell-1 of DAp) than in cells cultured at 22°C. Therefore, we confirm that P. subpacifica can produce DA, especially under P-limited conditions that could be associated with extraordinary oceanographic events such as the 2013–2016 "Blob" in the northeastern Pacific Ocean. This event altered local oceanographic conditions and possibly generated the presence of potential harmful species in areas with economic importance on the Mexican Pacific coast.

[1]  Thomas J. White,et al.  PCR protocols: a guide to methods and applications. , 1990 .

[2]  J. Doyle,et al.  Isolation of plant DNA from fresh tissue , 1990 .

[3]  T. White Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics , 1990 .

[4]  R. Guillard,et al.  Stichochrysis immobilis is a diatom, not a chrysophyte , 1993 .

[5]  G. Hallegraeff Species of the Diatom Genus Pseudonitzschia in Australian Waters , 1994 .

[6]  N. Rabalais,et al.  Changes in nutrient structure of river-dominated coastal waters: stoichiometric nutrient balance and its consequences , 1995 .

[7]  A. Coleman,et al.  The Internal Transcribed Spacer 2 Exhibits a Common Secondary Structure in Green Algae and Flowering Plants , 1997, Journal of Molecular Evolution.

[8]  A. W. Coleman,et al.  The significance of a coincidence between evolutionary landmarks found in mating affinity and a DNA sequence. , 2000, Protist.

[9]  M. Busman,et al.  Pseudo-nitzschia sp. cf. pseudodelicatissima — a confirmed producer of domoic acid from the northern Gulf of Mexico , 2001 .

[10]  N. Adams,et al.  MORPHOLOGICAL, TOXICOLOGICAL, AND GENETIC DIFFERENCES AMONG PSEUDO‐NITZSCHIA (BACILLARIOPHYCEAE) SPECIES IN INLAND EMBAYMENTS AND OUTER COASTAL WATERS OF WASHINGTON STATE, USA 1 , 2002 .

[11]  G. Fryxell,et al.  Morphology, phylogeny and taxonomy of species within the Pseudo-nitzschia americana complex (Bacillariophyceae) with descriptions of two new species, Pseudo-nitzschia brasiliana and Pseudo-nitzschia linea , 2002 .

[12]  S. G. Lee,et al.  Monthly Monitoring of Domoic Acid Producer Pseudo-nitzschia multiseries (Hasle) Hasle Using Species-Specific DNA Probes and WGA Lectins and Abundance of Pseudo-nitzschia Species (Bacillariophyceae) from Chinhae Bay, Korea , 2002 .

[13]  G. Hasle Are most of the domoic acid-producing species of the diatom genus Pseudo-nitzschia cosmopolites? , 2002 .

[14]  G. Hasle,et al.  A STUDY OF THE PSEUDO‐NITZSCHIA PSEUDODELICATISSIMA/CUSPIDATA COMPLEX (BACILLARIOPHYCEAE): WHAT IS P. PSEUDODELICATISSIMA? 1 , 2003 .

[15]  John P. Huelsenbeck,et al.  MrBayes 3: Bayesian phylogenetic inference under mixed models , 2003, Bioinform..

[16]  D. Hernández-Becerril Species of the planktonic diatom genus Pseudo-nitzschia of the Pacific coasts of Mexico , 1998, Hydrobiologia.

[17]  C. Cusack,et al.  Occurrence of Species from the Genus Pseudo-nitzschia Peragallo in Irish Waters , 2022, Biology and Environment: Proceedings of the Royal Irish Academy.

[18]  J. Ehrman,et al.  Diversity of the diatom genus Pseudo-nitzschia Peragallo in the Quoddy Region of the Bay of Fundy, Canada. , 2005 .

[19]  Thomas Dandekar,et al.  Homology modeling revealed more than 20,000 rRNA internal transcribed spacer 2 (ITS2) secondary structures. , 2005, RNA.

[20]  Tobias Müller,et al.  4SALE – A tool for synchronous RNA sequence and secondary structure alignment and editing , 2006, BMC Bioinformatics.

[21]  C. Lange,et al.  A review ofPseudo-nitzschia, with special reference to the Skagerrak, North Atlantic, and adjacent waters , 1996, Helgoländer Meeresuntersuchungen.

[22]  Tobias Müller,et al.  The internal transcribed spacer 2 database—a web server for (not only) low level phylogenetic analyses , 2006, Nucleic Acids Res..

[23]  Annette W. Coleman,et al.  Pan-eukaryote ITS2 homologies revealed by RNA secondary structure , 2007, Nucleic acids research.

[24]  É. Nézan,et al.  France : Identification of Pseudo-nitzschia multistriata and P. subpacifica from French waters. Were they part of the cryptic flora? , 2007 .

[25]  A. Amato,et al.  Reproductive isolation among sympatric cryptic species in marine diatoms. , 2007, Protist.

[26]  A. Amato,et al.  Morphology, phylogeny, and sexual cycle of Pseudo-nitzschia mannii sp. nov. (Bacillariophyceae): a pseudo-cryptic species within the P. pseudodelicatissima complex , 2008 .

[27]  Katherine Hubbard,et al.  IDENTIFICATION AND ASSESSMENT OF DOMOIC ACID PRODUCTION IN OCEANIC PSEUDO‐NITZSCHIA (BACILLARIOPHYCEAE) FROM IRON‐LIMITED WATERS IN THE NORTHEAST SUBARCTIC PACIFIC 1 , 2008, Journal of phycology.

[28]  Tobias Müller,et al.  The ITS2 Database II: homology modelling RNA structure for molecular systematics , 2007, Nucleic Acids Res..

[29]  S. Bates,et al.  Analysis of trace levels of domoic acid in seawater and plankton by liquid chromatography without derivatization, using UV or mass spectrometry detection. , 2009, Journal of chromatography. A.

[30]  Yann Ponty,et al.  VARNA: Interactive drawing and editing of the RNA secondary structure , 2009, Bioinform..

[31]  G. Casteleyn,et al.  DIVERSITY AND ABUNDANCE OF POTENTIALLY TOXIC PSEUDO-NITZSCHIA PERAGALLO IN AVEIRO COASTAL LAGOON, PORTUGAL AND DESCRIPTION OF A NEW VARIETY, P. PUNGENS VAR. AVEIRENSIS VAR. NOV. , 2009 .

[32]  A. Coleman Is there a molecular key to the level of "biological species" in eukaryotes? A DNA guide. , 2009, Molecular phylogenetics and evolution.

[33]  A. Amato,et al.  Intracellular domoic acid production in Pseudo-nitzschia multistriata isolated from the Gulf of Naples (Tyrrhenian Sea, Italy). , 2010, Toxicon : official journal of the International Society on Toxinology.

[34]  C. Cusack,et al.  Microscopic and molecular methods for quantitative phytoplankton analysis , 2010 .

[35]  Alexander Keller,et al.  The ITS2 Database III—sequences and structures for phylogeny , 2009, Nucleic Acids Res..

[36]  S. Quijano-Scheggia,et al.  Seasonal diversity of Pseudo-nitzschia sp. in the central zone of the Mexican Tropical Pacific, Manzanillo, Colima , 2010 .

[37]  Guadalupe Talavera Distribución espacial y temporal de diatomeas del género Pseudo-nitzschia spp y la concentración de ácido domoico en la región de la Bahía de Todos Santos, B.C., México , 2010 .

[38]  E. García-Mendoza,et al.  Growth and domoic acid content of Pseudo-nitzschia australis isolated from northwestern Baja California, Mexico, cultured under batch conditions at different temperatures and two Si:NO3 ratios , 2011 .

[39]  Klaus Peter Schliep,et al.  phangorn: phylogenetic analysis in R , 2010, Bioinform..

[40]  S. Quijano-Scheggia,et al.  First report of Pseudo-nitzschia brasiliana and P. micropora (Bacillariophycea) found in Cuyutlan Lagoon, Mexico , 2011 .

[41]  S. Bates,et al.  Pseudo-nitzschia physiological ecology, phylogeny, toxicity, monitoring and impacts on ecosystem health , 2012 .

[42]  N. Lundholm,et al.  Diversity, taxonomy and biogeographical distribution of the genus Pseudo-nitzschia (Bacillariophyceae) in Guangdong coastal waters, South China Sea , 2012 .

[43]  C. Leaw,et al.  MORPHOLOGY AND MOLECULAR CHARACTERIZATION OF PSEUDO‐NITZSCHIA (BACILLARIOPHYCEAE) FROM MALAYSIAN BORNEO, INCLUDING THE NEW SPECIES PSEUDO‐NITZSCHIA CIRCUMPORA SP. NOV , 2012, Journal of phycology.

[44]  C. Leaw,et al.  Genetic diversity of Pseudo-nitzschia brasiliana (Bacillariophyceae) from Malaysia , 2012, Journal of Applied Phycology.

[45]  S. Bates,et al.  Pseudo-nitzschia (Bacillariophyceae) species, domoic acid and amnesic shellfish poisoning: revisiting previous paradigms , 2012 .

[46]  Kimon K. Moschandreou,et al.  Inter- and intra-specific diversity of Pseudo-nitzschia (Bacillariophyceae) in the northeastern Mediterranean , 2012 .

[47]  M. Wolf,et al.  Compensatory Base Changes in ITS2 Secondary Structures Correlate with the Biological Species Concept Despite Intragenomic Variability in ITS2 Sequences – A Proof of Concept , 2013, PloS one.

[48]  C. Leaw,et al.  Three novel species in the Pseudo‐nitzschia pseudodelicatissima complex: P. batesiana sp. nov., P. lundholmiae sp. nov., and P. fukuyoi sp. nov. (Bacillariophyceae) from the Strait of Malacca, Malaysia , 2013, Journal of phycology.

[49]  S. Quijano-Scheggia,et al.  The genus Pseudo-nitzschia (Bacillariophyceae) in Manzanillo and Santiago Bays, Colima, Mexico , 2013 .

[50]  E. Orive,et al.  The genus Pseudo-nitzschia (Bacillariophyceae) in a temperate estuary with description of two new species: Pseudo-nitzschia plurisecta sp. nov. and Pseudo-nitzschia abrensis sp. nov. , 2013, Journal of phycology.

[51]  Tobias Müller,et al.  ITS2, 18S, 16S or any other RNA - simply aligning sequences and their individual secondary structures simultaneously by an automatic approach. , 2014, Gene.

[52]  Katherine Hubbard,et al.  Diversity and toxicity of the diatom Pseudo-nitzschia Peragallo in the Gulf of Maine, Northwestern Atlantic Ocean. , 2014, Deep-sea research. Part II, Topical studies in oceanography.

[53]  Diversity and seasonality of Pseudo-nitzschia (Peragallo) at two North Sea time-series monitoring sites , 2015, Helgoland Marine Research.

[54]  J. Schultz,et al.  ITS2 Database V: Twice as Much. , 2015, Molecular biology and evolution.

[55]  M. Wolf,et al.  A non‐toxigenic but morphologically and phylogenetically distinct new species of Pseudo‐nitzschia, P. sabit sp. nov. (Bacillariophyceae) , 2015, Journal of phycology.

[56]  D. L. Swain A tale of two California droughts: Lessons amidst record warmth and dryness in a region of complex physical and human geography , 2015 .

[57]  Nicholas A. Bond,et al.  Causes and impacts of the 2014 warm anomaly in the NE Pacific , 2015 .

[58]  R. Kudela,et al.  An unprecedented coastwide toxic algal bloom linked to anomalous ocean conditions , 2016, Geophysical research letters.

[59]  Ahmed El Mallahi,et al.  Digital holographic microscopy: a novel tool to study the morphology, physiology and ecology of diatoms , 2016 .

[60]  Irina Koester,et al.  Biological Impacts of the 2013–2015 Warm-Water Anomaly in the Northeast Pacific: Winners, Losers, and the Future , 2016 .

[61]  Cecilia Tenorio,et al.  Morphological and toxicological studies of Pseudo-nitzschia species from the central coast of Peru , 2016 .

[62]  C. Leaw,et al.  High diversity of Pseudo‐nitzschia along the northern coast of Sarawak (Malaysian Borneo), with descriptions of P. bipertita sp. nov. and P. limii sp. nov. (Bacillariophyceae) , 2016, Journal of phycology.

[63]  J. Fauchot,et al.  Inter and intra-specific growth and domoic acid production in relation to nutrient ratios and concentrations in Pseudo-nitzschia: phosphate an important factor. , 2017, Harmful algae.

[64]  Fei-xue Fu,et al.  Understanding the blob bloom: Warming increases toxicity and abundance of the harmful bloom diatom Pseudo-nitzschia in California coastal waters. , 2017, Harmful algae.

[65]  B. Moore,et al.  Biosynthesis of the neurotoxin domoic acid in a bloom-forming diatom , 2018, Science.

[66]  M. Wolf,et al.  Phylogeny and species delineation in the marine diatom Pseudo‐nitzschia (Bacillariophyta) using cox1, LSU, and ITS2 rRNA genes: A perspective in character evolution , 2018, Journal of phycology.

[67]  W. Grebner,et al.  Induction of defensive traits in marine plankton—new copepodamide structures , 2018, Limnology and Oceanography.

[68]  Lia Godinho,et al.  Evaluation of intracellular and extracellular domoic acid content in Pseudo-nitzschia multiseries cell cultures under different light regimes. , 2018, Toxicon : official journal of the International Society on Toxinology.

[69]  Katherine Hubbard,et al.  Pseudo-nitzschia, Nitzschia, and domoic acid: New research since 2011. , 2018, Harmful algae.

[70]  Y. Li,et al.  Pseudo‐nitzschia nanaoensis sp. nov. (Bacillariophyceae) from the Chinese coast of the South China Sea , 2018, Journal of phycology.

[71]  Elise Van Meerssche,et al.  Coastal eutrophication and freshening: Impacts on Pseudo-nitzschia abundance and domoic acid allelopathy , 2018, Estuarine, Coastal and Shelf Science.

[72]  S. Quijano-Scheggia,et al.  Description of Pseudo-nitzschia cuspidata var. manzanillensis var. nov. (Bacillariophyceae): morphology and molecular characterization of a variety from the central Mexican pacific , 2018 .

[73]  Ø. Moestrup,et al.  Morphological, molecular and toxigenic characteristics of Namibian Pseudo-nitzschia species - including Pseudo-nitzschia bucculenta sp. nov. , 2018, Harmful algae.

[74]  J. Cáceres‐Martínez,et al.  Mass Mortality of Cultivated Northern Bluefin Tuna Thunnus thynnus orientalis Associated With Chattonella Species in Baja California, Mexico , 2018, Front. Mar. Sci..

[75]  S. Murray,et al.  A new diatom species P. hallegraeffii sp. nov. belonging to the toxic genus Pseudo-nitzschia (Bacillariophyceae) from the East Australian Current , 2018, PloS one.

[76]  Sudhir Kumar,et al.  MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. , 2018, Molecular biology and evolution.

[77]  U. John,et al.  Induction of domoic acid production in diatoms-Types of grazers and diatoms are important. , 2018, Harmful algae.

[78]  N. Lundholm,et al.  Copepods drive large-scale trait-mediated effects in marine plankton , 2019, Science Advances.

[79]  A. Amato,et al.  Cryptic Diversity: a Long-lasting Issue for Diatomologists. , 2019, Protist.