Abstract

Dense patches were observed in the tidal pools of the southern area of Korea. To clarify the causative organisms, the cells were collected and their morphological features were examined using light and scanning electron microscopy (SEM). In addition, after establishing strains for the cells the molecular phylogeny was inferred with concatenated small subunit (SSU) and large subunit (LSU) rRNA sequences. The cells were characterized by a nucleus in the hypotheca, strong reticulations in thecal plates, the separation of plates 2a and 3a, the tear-shaped apical pore complex, an elongated rectangular 1a plate and the absence of the right sulcal list. The thecal plate formula was Po, X, 4′, 3a, 7″, 6c, 4S, 5′′′, 2′′′′. Based on these morphological features, the cells were identified as Bysmatrum subsalsum. In the culture, the spherical cysts of B. subsalsum without thecal plates were observed. Molecular phylogeny revealed two ribotypes of B. subsalsum are identified; The Korean isolates were nested within the ribotype B consisting of the isolates from China, Malaysia and the French Atlantic, whereas the ribotype A includes only the isolates from the Mediterranean Sea. In the phylogeny, B. subsalsum and B. austrafrum were grouped. This can be supported by the morphological similarity between the two species, indicating that the two species may be conspecific, however B. subsalsum may distinguish from B. austrafrum, because of differences in the types of eyespots reported in previous studies. These findings support the idea that there is cryptic diversity within B. subsalsum.

Highlights

  • Luo et al [8] documented that B. subsalsum strains from Malaysia and the French Atlantic formed a subclade that can be distinguished from a clade consisting of stains from the Mediterranean Sea and that there is cryptic diversity within B. subsalsum, based on the genetic distance shown in ITS sequences among Bysmatrum species [7]

  • We describe the morphological details of Korean strains of B. subsalsum and report on their molecular characterization, based on concatenated small subunit (SSU) and large subunit (LSU) rRNA gene sequences

  • A yellowish eyespot was located on the right side of the sulcus (Figure 1d,e)

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Summary

Introduction

Luo et al [8] documented that B. subsalsum strains from Malaysia and the French Atlantic formed a subclade (ribotype B) that can be distinguished from a clade (ribotype A) consisting of stains from the Mediterranean Sea and that there is cryptic diversity within B. subsalsum, based on the genetic distance shown in ITS sequences among Bysmatrum species [7] This indicates that additional molecular data (including morphological descriptions) are needed to clarify the cryptic diversity of B. subsalsum, with strains established from water bodies of various geographical regions. We describe the morphological details of Korean strains of B. subsalsum and report on their molecular characterization, based on concatenated small subunit (SSU) and large subunit (LSU) rRNA gene sequences

Sampling and Culture
Morphological Observation
DNA Extraction and Sequencing
Phylogenetic Analysis
Morphology of Vegetative Cell and Resting Cyst of Bysmatrum subsalsum
Morphology of Bysmatrum subsalsum Cyst
Phylogenetic Position of Bysmatrum subsalsum
Environmental Conditions in Relation to the Growth of Bysmtrum subsalsum
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