Molecular phylogeny and ultrastructure of two novel parasitic dinoflagellates, Haplozoon gracile sp. nov. and ​H. pugnus sp. nov.

Abstract

ABSTRACT This study describes two novel parasitic dinoflagellates: Haplozoon gracile sp. nov. isolated from a bamboo worm (Maldanidae), ‘cf. Petaloclymene sp.’ sensu Kobayashi et al. 2018; and, H. pugnus sp. nov. isolated from Nicomache sp. and Nicomache personata (Maldanidae). Trophonts (feeding stages) were observed with light, scanning, and transmission electron microscopy. Molecular phylogenetic analyses were performed based on 18S rDNA. COI sequences were obtained for host organisms. Trophonts of H. gracile were linear (single longitudinal row) and relatively slender with a mean length of 190 μm, and consisted of a long and narrow trophocyte, rectangular gonocytes (mean width = 10 μm), and slightly rounded sporocytes. Trophonts of H. pugnus were pectinate (1–8 rows of sporocytes in one plane), with a mean length of 179 μm, consisting of a bulbous trophocyte, rectangular gonocytes (mean width = 25 μm), and rounded sporocytes. The body of both species was covered with many depressions that overlaid the amphiesmal vesicles. TEM observations of trophocytes in H. gracile revealed a stylet with a central dense core and rich mitochondria subtending the amphiesma. Furthermore, amphiesmal vesicles appeared to contain thecal plates in both species. Phylogenetic analyses generally resolved a Haplozoon clade, and H. gracile and H. pugnus were clearly distinguished from other species for which molecular data are available. Based on the morphological and host comparisons with all described species and their molecular phylogeny, we conclude that these two isolates are new species of Haplozoon, H. gracile sp. nov. and H. pugnus sp. nov.