Genetic relatedness of toxic and nontoxic isolates of the marine pennate diatom Pseudonitzschia (Bacillariophyceae): phylogenetic analysis of 18S rRNA sequences.

The nuclear small subunit (SSU) rRNA genes from several marine diatoms, including 2 species that have been responsible for toxic blooms, were amplified from total DNA by the polymerase chain reaction (PCR), and the sequences analysed to determine their genetic relatedness. The isolates investigated include 2 morphologically similar forms of the pennate diatom Pseudonitzschia pungens: (1) P. pungens f. multiseries, a known producer of the toxin domoic acid, and (2) P. pungens f. pungens, which is not toxic. Strains of a second toxin-producing species, P. australis, and a nontoxic Thalassiosira species were also included in this study. Phylogenetic analyses of sequences by both distance and parsimony methods clearly distinguished the 2 forms of P. pungens and the 2 Pseudonitzschia species from other diatoms for which sequence data are available. Differences in the nucleotide sequences of the 2 forms of P. pungens permitted the design of PCR primers that allowed discrimination between them. This may prove a valuable tool in identifying toxic and nontoxic forms of closely related and morphologically similar diatom species.

[1]  D. Bhattacharya,et al.  ALGAE CONTAINING CHLOROPHYLLS a + c ARE PARAPHYLETIC: MOLECULAR EVOLUTIONARY ANALYSIS OF THE CHROMOPHYTA , 1992, Evolution; international journal of organic evolution.

[2]  M. Ragan,et al.  Phylogenetic relationships in the Gracilariales (Rhodophyta) as determined by 18S rDNA sequences , 1992 .

[3]  D. Garrison,et al.  CONFIRMATION OF DOMOIC ACID PRODUCTION BY PSEUDONITZSCHIA AUSTRALIS (BACILLARIOPHYCEAE) CULTURES 1 , 1992 .

[4]  M. Quilliam,et al.  AN OUTBREAK OF DOMOIC ACID POISONING ATTRIBUTED TO THE PENNATE DIATOM PSEUDONITZSCHIA AUSTRALIS 1 , 1992 .

[5]  D. Mann,et al.  ON THE REPORT OF GAMETOGENESIS, OOGAMY, AND UNIFLAGELLATED SPERM IN THE PENNATE DIATOM NITZSCHIA PUNGENS (1991. J. PHYCOL. 27:21–26) , 1992 .

[6]  G. L. Floyd,et al.  Ultrastructure of the flagellar apparatus of the gametes of Gongrosira papuasica (Chlorophyta) , 1992 .

[7]  L. Medlin,et al.  MORPHOLOGICAL AND GENETIC VARIATION WITHIN THE DIATOM SKELETONEMA COSTATUM (BACILLARIOPHYTA): EVIDENCE FOR A NEW SPECIES, SKELETONEMA PSEUDOCOSTATUM 1 , 1991 .

[8]  F. Partensky,et al.  FLOW CYTOMETRY AND MICROSCOPY OF GAMETOGENESIS IN NITZSCHIA PUNGENS, A TOXIC, BLOOM‐FORMING, MARINE DIATOM 1 , 1991 .

[9]  R. Amann,et al.  Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations , 1990, Applied and environmental microbiology.

[10]  L. Medlin COMPARISON OF RESTRICTION ENDONUCLEASE SITES IN THE SMALL SUBUNIT 16S-like rRNA GENE FROM THE MAJOR GENERA OF THE FAMILY BACILLARIACEAE , 1990 .

[11]  D. Tindall,et al.  GENITIC VARIABILITY IN TOXIN POTENCIES AMONG SEVENTEEN CLONES OF GAMBIERDISCUS TOXICUS (DINOPHYCEAE) 1 , 1989 .

[12]  S. Bates,et al.  Pennate Diatom Nitzschia pungens as the Primary Source of Domoic Acid, a Toxin in Shellfish from Eastern Prince Edward Island, Canada , 1989 .

[13]  E. Delong,et al.  Phylogenetic stains: ribosomal RNA-based probes for the identification of single cells. , 1989, Science.

[14]  C. Marck,et al.  'DNA Strider': a 'C' program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers. , 1988, Nucleic acids research.

[15]  K. Mullis,et al.  Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. , 1988, Science.

[16]  N. Saitou,et al.  The neighbor-joining method: a new method for reconstructing phylogenetic trees. , 1987, Molecular biology and evolution.

[17]  R. Lande,et al.  QUANTITATIVE GENETIC ANALYSIS OF MORPHOLOGICAL VARIATION IN AN ANTARCTIC DIATOM GROWN AT TWO LIGHT INTENSITIES 1 , 1987 .

[18]  M. Sogin,et al.  Phylogenetic evidence for the acquisition of ribosomal RNA introns subsequent to the divergence of some of the major Tetrahymena groups. , 1986, The EMBO journal.

[19]  J. Felsenstein CONFIDENCE LIMITS ON PHYLOGENIES: AN APPROACH USING THE BOOTSTRAP , 1985, Evolution; international journal of organic evolution.

[20]  D Sankoff,et al.  On the evolutionary descent of organisms and organelles: a global phylogeny based on a highly conserved structural core in small subunit ribosomal RNA. , 1984, Nucleic acids research.

[21]  S. Garger,et al.  Rapid purification of plasmid DNA by a single centrifugation in a two-step cesium chloride-ethidium bromide gradient. , 1983, Biochemical and biophysical research communications.

[22]  G. Robinson,et al.  Electrophoretic analysis of the species and population structure of the diatom Asterionella formosa , 1983 .

[23]  R. Guillard,et al.  Genetic variability and differentiation in the temperature niche component of the diatom Thalassiosira pseudonana , 1981 .

[24]  J. C. Gallagher POPULATION GENETICS OF SKELETONEMA COSTATUM (BACILLARIOPHYCEAE) IN NARRAGANSETT BAY 1 , 1980 .

[25]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[26]  D. Anderson,et al.  Discrimination between domoic-acid-producing and nontoxic forms of the diatom Pseudonitzschia pungens using immunofluorescence , 1993 .

[27]  G. Hasle Nomenclatural notes on marine planktonic diatoms. The family Bacillariaceae , 1993 .

[28]  F. Chavez,et al.  Autecology of the diatom Pseudonitzschia australis, a domoic acid producer, from Monterey Bay, California , 1992 .

[29]  D. J. Douglas,et al.  Production of Domoic Acid, a Neurotoxic Amino Acid, by an Axenic Culture of the Marine Diatom Nitzschia pungens f. multiseries Hasle , 1992 .

[30]  N. Pace,et al.  Microbial ecology and evolution: a ribosomal RNA approach. , 1986, Annual review of microbiology.

[31]  R. Guillard,et al.  Culture of Phytoplankton for Feeding Marine Invertebrates , 1975 .