A New Ribo-Type of Wangodinium sinense from Germination of Resting Cysts Isolated from Ballast Tank Sediments of Incoming Ships to China

Abstract

In recent decades, ships’ ballast water and associated sediments have been recognized globally as significant vectors for the dissemination of non-indigenous species, which has attracted extensive attention due to its ecological and economic impacts. The characteristics of production of resting cysts in the dinoflagellate life cycle further increases the risk of biological invasions through ballast tank sediments. Despite extensive research which has characterized the species diversity of dinoflagellate cysts within ballast tank sediments, the possibility and importance of invasions caused by different ribosomal types of the same species have been paid little attention. In this study, two cultures of dinoflagellates were established through cyst germination from the ballast tank sediments collected from two ships (“THETIS” and “WARIYANAREE”) arriving at the Jiangyin Port (China) and identified as Wangodinium sinense Z. Luo, Zhangxi Hu, Yingzhong Tang and H.F. Gu by comprehensive phylogenetic analysis of rDNA sequences (including LSU, SSU, and ITS1-5.8S-ITS2). Despite the rDNA sequences of the isolates showing a generally high similarity to reference sequences, the LSU D1-D6 sequences contained up to 11 stable single nucleotide polymorphisms (SNPs), while SSU and ITS1-5.8S-ITS2 sequences exhibited up to five and two divergence sites, respectively. Moreover, phylogenetic analyses based on partial LSU and SSU rDNA sequences further indicated that strains germinated from ships’ ballast tank sediments formed a strongly supported sister clade to the strains previously isolated from Chinese and Korean waters, representing a novel ribo-type distinct from Chinese and Korean strains. Detailed morphological observations using light microscopy (LM) and scanning electron microscopy (SEM) did not find differences between our isolates and the holotype of the species in key diagnostic characteristics including the position and shape of the nucleus and chloroplasts, as well as the ASC structure, which suggested that no significant morphological divergence has occurred among these ribo-types. Acute toxicity exposure assays indicated that this ribo-type of W. sinense posed no lethal effect on rotifers at concentrations ≤ 104 cells/mL, yet it remains necessary to maintain vigilance regarding the potential risk of algal blooms resulting from higher cell density or environmental changes in the invaded ecosystems. This study reports the first successful germination of W. sinense cysts from ballast tank sediments, indicating that its cysts may be widely transferred through ballast tank sediments, and presents a potential risk of bio-invasions of new genotypes of species to a region where other genotypes of the same species have been present as indigenous species.

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