ABSTRACT. We determined 18S rRNA gene sequences of Amoebophrya strains infecting the thecate dinoflagellates Alexandrium affine and Gonyaulax polygramma from Korean coastal waters and compared those data with previously reported sequences of Amoebophrya from cultures, infected cells concentrated from field samples, and environmental 18S rRNA gene sequences obtained from a variety of marine environments. Further, we used these data to examine genetic diversity in Amoebophrya strains relative to geographic origin, host phylogeny, site of infection, and host specificity. In our analyses of known dinoflagellate taxa, the 13 available Amoebophrya sequences clustered together within the dinoflagellates as three groups forming a monophyletic group with high bootstrap support (maximum likelihood, ML: 100%) or a posterior probability (PP) of 1. When the Amoebophrya sequences were analyzed along with environmental sequences associated with Marine Alveolate Group II, nine subgroups formed a monophyletic group with high bootstrap support (ML: 100%) and PP of 1. Sequences known to be from Amoebophrya spp. infecting dinoflagellate hosts were distributed in seven of those subgroups. Despite differences in host species and geographic origin (Korea, United States, and Europe), Amoebophrya strains (Group II) from Gymnodinium instriatum, A. affine, Ceratium tripos (AY208892), Prorocentrum micans, and Ceratium lineatum grouped together by all of our tree construction methods, even after adding the environmental sequences. By contrast, strains within Groups I and III divided into several lineages following inclusion of environmental sequences. While Amoebophrya strains within Group II mostly developed within the host cytoplasm, strains in Groups I and III formed infections inside the host nucleus, a trait that appeared across several of the subgroups. Host specificity varied from moderately to extremely species-specific within groups, including Group II. Taken together, our results imply that genetic diversity in Amoebophrya strains does not always reflect parasite biology or biogeography.
Twenty-nine culture strains belonging to the genus Alexandrium Halim (Dinophyceae) were established from water column or sediments in Korea. Seventeen isolates were identified as A. tamarense (Lebour) Balech, eight isolates as A. sp. cf. catenella and one as A. catenella (Whedon et Kofoid) Balech according to the presence or absence of a ventral pore, the shape of the posterior sulcal plate and the sulcal width. Three isolates were unable to be identified due to considerable distortion of thecal plates and lack of enough materials, but typical of A. tamarense and/or A. catenella. The overall cell shape of A. tamarense was usually longer than wide. The posterior sulcal plate was definitely longer than wide dorsoventrally, and sulcus extended posteriorly without apparent widening. They were distributed in three major coasts of Korea. In contrast, the cell shape of A. sp. cf. catenella was generally anterior-posteriorly flattened. The transversal axis of the posterior sulcal plate was always longer than the longitudinal, or both axes were nearly equal in length. Its sulcus was broader than that of A. tamarense and widened in the direction of antapex about 1.5 times. This morphotype existed in nearshore and offshore waters of the southern Korea sea. One of A. catenella isolates from Jinhae Bay showed no conspicuous differences with A. sp. cf. catenella except for the consistent absence of a ventral pore.
Genetic variation in the Asian shore crab Hemigrapsus sanguineus was determined from partial mitochondrial DNA (mtDNA) sequences of the cytochrome b (Cytb) gene. Samples included 143 crabs from six localities along three coastlines in South Korea. A nucleotide sequence analysis revealed 38 variable sites in a 470-bp sequence, which defined 37 haplotypes. The haplotypes were not associated geographically and had a shallow genealogy. Pairwise F ST tests and a two-dimensional scaling analysis revealed no significant genetic differentiation among most of the populations. The low pairwise comparison values, but significant genetic differentiation of a northeastern population from all other populations, might have been influenced by a restriction in gene flow caused by hydrographic conditions such as ocean boundaries. The high haplotype diversity, low nucleotide diversity, and time since H. sanguineus expansion in Korean coastal waters indicate rapid population growth and a recent, sudden expansion in the Late Pleistocene.
Partial large subunit ribosomal DNA (LSU rDNA) sequences of Dissodinium pseudolunula Swift ex Elbrächter et Drebes, Pheopolykrikos hartmannii (Zimmermann) Matsuoka et Fukuyo, Polykrikos cf. schwartzii Bütschli and Polykrikos kofoidii Chatton were analyzed to reveal their phylogenetic status. The four athecate dinoflagellate species fell within the clade of Gymnodinium sensu G. Hansen et Moestrup supported by high likelihood values without apparent phylogenetic relationships to neither Gymnodinium species nor each other. Their genetic affiliations to typical Gymnodinium species were morphologically supported by the loop-shaped apical grooves in Pheopolykrikos and Polykrikos species and the Gymnodinium-like motile stage at least once during pleomorphic life cycles in D. pseudolunula and Ph. hartmannii.
Complementary DNA and genomic sequences representing two different beta-actins were isolated from a threatened freshwater fish species Hemibarbus mylodon. The beta-actin 1 and 2 encoded an identical number of amino acids (375 aa), and shared 88.8 and 99.7% of identity at coding nucleotide and amino acid levels, respectively. Genomic open reading frame (ORF) sequences of both isoforms contained five translated exons interrupted by four introns with conserved GT/AG exon/intron boundary rule. Semi-quantitative RT-PCR showed that the two isoform mRNAs were ubiquitously detected in all tissues tested, but transcript levels were variable across tissues. Phylogenetic analysis showed that H. mylodon beta-actin 1 and 2 were clustered into two distinct major and minor branches of Cypriniformes, respectively. Comparisons of the 5'-upstream region and 3'-UTR of H. mylodon beta-actin 1 also showed a high degree of homology with those of the major teleost beta-actins and warmblooded vertebrate beta-cytoskeletal actins, suggesting their more recent common origin.
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