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Identification of Pfiesteria Piscicida (Dinophyceae) and Pfiesteria ‐Like Organisms Using its‐Specific PCR Assays
Author(s) -
Litaker R. W.,
Vandersea M. W.,
Kibler S. R.,
Tester P. A.,
Reece K. S.,
Stokes N. A.,
Steidinger K. A.,
Millie D. F.,
Bendis B. J.,
Pigg R. J.
Publication year - 2003
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1111/j.0022-3646.2003.03906001_104.x
Subject(s) - biology , dinoflagellate , dinophyceae , ribosomal rna , internal transcribed spacer , algal bloom , primer (cosmetics) , botany , gene , genetics , ecology , phytoplankton , chemistry , organic chemistry , nutrient
The putative harmful algal bloom dinoflagellate, Pfiesteria piscicida , frequently co‐occurs with other morphologically similar species collectively known as Pfiesteria ‐like organisms (PLOs). This study specifically evaluated whether unique sequences in the ribosomal internal transcribed spacer regions (ITS1 and ITS2) could be used to develop PCR assays capable of detecting PLOs in natural assemblages. ITS regions were selected because they are more variable than the flanking small subunit (SSU) or large subunit (LSU) ribosomal RNA genes and more likely to contain species‐specific sequences. Sequencing of the ITS regions revealed unique oligonucleotide primer binding sites for Pfiesteria piscicida, Pfiesteria shumwayae , Florida “Lucy” species, two cryptoperidiniopsoid species, “H/V14” and “PLO21,” and the estuarine mixotroph, Karlodinium micrum . These PCR assays had a minimum sensitivity of 100 cells in a 100 mL sample (1 cell mL‐1) and were successfully used to detect PLOs in the St. Johns River system in Florida, USA.