Protoceratium reticulatum (Dinophyceae) in the austral Southwestern Atlantic and the first report on YTX-production in shelf waters of Argentina

Growth and bioactive secondary metabolites of arctic Protoceratium reticulatum (Dinophyceae)

“…In the present ML tree, all P. reticulatum were part of a monophyletic clade that subdivided into two well supported, divergent sub-clades. The sequences of the isolates fell in the same clade as the majority of sequences of P. reticulatum (''Clade A'' according to Akselman et al, 2015), and only three isolates of P. reticulatum comprised the second sub-clade within P. reticulatum (''Clade B'' according to Akselman et al, 2015, Fig. 3).…”

“…Genomic DNA was extracted from cell pellets using a DNeasy Plant Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions as described in detail by Toebe et al (2013). From the resultant DNA the D1/D2 hypervariable region of the large sub-unit (LSU) of the ribosomal operon was PCR amplified and Sanger sequenced with the primers D1R (forward) and D2C (reverse) (Scholin et al, 1994) and PCR chemistry, cycling and sequencing conditions as in Akselman et al (2015). Forward and reverse sequences were assembled in Geneious Pro 5.4.4 (Biomatters Ltd., Auckland, New Zealand) and a BLAST search was conducted with the resultant sequences in order to identify other P. reticulatum from GenBank.…”

“…Sequences of P. reticulatum and the resultant set of other P. reticulatum sequences were aligned in MAFFT v7.017 (Katoh and Kuma, 2002;Katoh and Standley, 2013) using a plugin for Geneious Pro. Additional sequences in the alignment corresponded to those gonyaulacoid dinoflagellates used in the phylogenetic analysis by Akselman et al (2015) that built up on a previous analysis in Howard et al (2009), but additionally included a set of sequences representing the five ribotypes of the Alexandrium tamarense/A. fundyense/A.…”

“…catenella species complex and a sequence of Prorocentrum minimum as the out-group (see Supplementary material, Table S01). The resultant alignment was restricted to the length of the sequences of the clonal isolates of P. reticulatum from Greenland and sequences of P. reticulatum ''Clade A'' (sensu Akselman et al, 2015) that were identical were removed from the alignment. The best fitting model of nucleotide substitution was determined according to scores of the Akaike Information Criterion in jModeltest 0.1.1 (Posada, 2008) and a Maximum Likelihood (ML) tree was constructed using the TIM1+G model of nucleotide substitution in PhyML (Guindon and Gascuel, 2003) via a plugin in Geneious Pro.…”

“…P. aceros, Gonyaulax grindleyi) is a gonyaulacoid dinoflagellate with a taxonomic and nomenclatural history dating back to 1858 when Claparade & Lachman first described the species as Peridinium reticulatum (Hansen et al, 1996/97). The species is fairly common and widely distributed in coastal waters around the world-although it is genetically subdivided into two evolutionary clades, for which distinct distribution ranges are currently known (see Akselman et al, 2015, and references therein). A main event gaining attention was when Protoceratium recticulatum was identified as a producer of yessotoxins (YTX), first confirmed in cultures of P. reticulatum from New Zealand (Satake et al, 1997).…”