Myrionecta rubra population genetic diversity and its cryptophyte chloroplast specificity in recurrent red tides in the Columbia River estuary

For several decades, annually recurring blooms of the photosynthetic ciliate Myrionecta rubra have been observed in the Columbia River estuary in late summer. In an effort to understand the dynamics of these blooms, we investigated the genetic variability of M. rubra and its cryptophyte plastids within 3 large estuarine blooms formed in consecutive years (2007 to 2009), and conducted a broader spatial survey along the coasts of Oregon and Washington. Analysis of the '18S-28S' sequences specific for Mesodiniidae uncovered at least 5 variants of M. rubra within the Columbia River coastal margin in spring and summer, but only one of these M. rubra variants was implicated in estuary bloom formation. Using a multigene approach, we show that the bloom-forming variant of M. rubra appears to harbor the same cryptophyte chloroplast in recurring blooms. Analyses of chloro- plast 16S rRNA, cryptophyte RuBisCO and Photosystem II D2 genes together suggest that the plastid is derived from Teleaulax amphioxeia. Free-living cells of this species and of other cryptophytes were practically absent from the bloom patches in the estuary main channels based on 18S rDNA sequence analyses. The respectively low and high proportions of T. amphioxeia nuclei and chloroplast signals found in the M. rubra bloom of the Columbia River estuary in successive years supports the notion of an association (either endosymbiosis or kleptoplastidy) between T. amphioxeia and the bloom-form- ing M. rubra variant, with loss of cryptophyte nuclei. The genetic variability of M. rubra uncovered here is relevant to the controversy in the literature regarding the cryptophyte/M. rubra association.