Morphological, phylogenetic and metabolite profile of <i>Prorocentrum clipeus</i>, a newly recorded epiphytic dinoflagellate in the northern Yellow Sea

Ruifang Wang; Mengmeng Tong; Shiwen Zhou; Junjie Zheng; Wenguang Zhang; Xinfeng Dai; Douding Lu; Jiarong Hu; Tianze Leng; Qinglin Mu; Zhongyong Yan; Jiangning Zeng; Pengbin Wang

doi:10.1007/s13131-024-2302-2

Volume 43 Issue 8

Aug. 2024

Turn off MathJax

Article Contents

Article Navigation > Acta Oceanologica Sinica > 2024 > 43(8): 128-141

Ruifang Wang, Mengmeng Tong, Shiwen Zhou, Junjie Zheng, Wenguang Zhang, Xinfeng Dai, Douding Lu, Jiarong Hu, Tianze Leng, Qinglin Mu, Zhongyong Yan, Jiangning Zeng, Pengbin Wang. Morphological, phylogenetic and metabolite profile of Prorocentrum clipeus, a newly recorded epiphytic dinoflagellate in the northern Yellow Sea[J]. Acta Oceanologica Sinica, 2024, 43(8): 128-141. doi: 10.1007/s13131-024-2302-2

Citation:

Ruifang Wang, Mengmeng Tong, Shiwen Zhou, Junjie Zheng, Wenguang Zhang, Xinfeng Dai, Douding Lu, Jiarong Hu, Tianze Leng, Qinglin Mu, Zhongyong Yan, Jiangning Zeng, Pengbin Wang. Morphological, phylogenetic and metabolite profile of Prorocentrum clipeus, a newly recorded epiphytic dinoflagellate in the northern Yellow Sea[J]. Acta Oceanologica Sinica, 2024, 43(8): 128-141. doi: 10.1007/s13131-024-2302-2

Citation:

Ruifang Wang, Mengmeng Tong, Shiwen Zhou, Junjie Zheng, Wenguang Zhang, Xinfeng Dai, Douding Lu, Jiarong Hu, Tianze Leng, Qinglin Mu, Zhongyong Yan, Jiangning Zeng, Pengbin Wang. Morphological, phylogenetic and metabolite profile of Prorocentrum clipeus, a newly recorded epiphytic dinoflagellate in the northern Yellow Sea[J]. Acta Oceanologica Sinica, 2024, 43(8): 128-141. doi: 10.1007/s13131-024-2302-2

PDF( 6398 KB)

Morphological, phylogenetic and metabolite profile of Prorocentrum clipeus, a newly recorded epiphytic dinoflagellate in the northern Yellow Sea

doi: 10.1007/s13131-024-2302-2

1.
Ocean College, Zhejiang University, Zhoushan 316021, China
2.
Key Laboratory of Marine Ecosystem Dynamics, Second Institute of Oceanography, Ministry of Natural Resources, Hangzhou 310012, China
3.
The State Key Laboratory of Marine Pollution, Department of Biomedical Sciences, City University of Hong Kong, Hong Kong SAR 999077, China
4.
Zhejiang Marine Ecology and Environment Monitoring Center, Zhoushan 316000, China

Funds: The National Natural Science Foundation of China under contract Nos 41706191 and 41961144013; the Natural Science Foundation of Zhejiang Province under contract No. LY20D060004; the National Natural Science Foundation of China under contract Nos 41676111, 41876139 and 41906140; the Program of Bureau of Science and Technology of Zhoushan Grant under contract No. 2019C81031; the Basic Public Welfare Research Project of Zhejiang Province under contract No. LGC22B050032.

More Information

Corresponding author: jiangningz@126.com; algae@sio.org.cn
Received Date: 2023-10-04
Accepted Date: 2024-02-29

Available Online: 2024-07-10

Publish Date: 2024-08-25

Abstract

Abstract

More than 30 species of benthic Prorocentrum have been identified, some of which produce okadaic acid (OA) and its derivatives, dinophysistoxins (DTXs), which cause diarrhetic shellfish poisoning (DSP). Increasing numbers of benthic Prorocentrum species have been reported in tropical and subtropical waters of China. In contrast, only a few benthic Prorocentrum species have been reported in temperate waters. In this study, morphological descriptions obtained using light microscopy, scanning electron microscopy and molecular characterization of one Prorocentrum clipeus strain isolated from the Yellow Sea are presented. Prorocentrum clipeus cells were nearly circular in shape, with a collar, ridge, and one protrusion. The periﬂagellar area was wide U-shaped, with two curved projections on platelet 1a. Nine periﬂagellar platelets of different sizes were observed. The morphology closely ﬁts that of the species isolated from other locations. Phylogenetic analysis based on the molecular sequences of the small subunit (SSU) rDNA, internal transcribed spacer (ITS), and large subunit (LSU) rDNA was performed. A comprehensive metabolomic analysis incorporating target, suspect and non-target screenings was first applied to investigate the intracellular and extracellular metabolite profiles of the current isolate of P. clipeus. According to the results of the target and suspect screenings, 179 metabolites or toxins produced by DSP-related algal species, including OA, dinophysistoxin-1 (DTX1), dinophysistoxin-2 (DTX2) and pectenotoxin-2 (PTX2), were not detected. Non-target screening involving feature-based molecular networking (FBMN) provided a global view of major metabolites produced by the P. clipeus DF128 strain and revealed 23 clusters belonging to at least 13 compound classes, with organometallic compounds, lipids and lipid-like molecules, phenylpropanoids and polyketides, and benzenoids as major types. To date, this is the ﬁrst record of the characterization of P. clipeus in samples from Chinese waters. Our results support the wide distribution of epibenthic Prorocentrum species.
- Prorocentrum,
- taxonomy,
- phylogeny,
- metabolite profiling,
- Yellow Sea

# Ruifang Wang and Mengmeng Tong contributed equally to this work.

FullText(HTML)

References(63)

References

Andersen R A. 2005. Algal Culturing Techniques. Oxford: Elsevier Academic Press, 578

Arteaga-Sogamoso E, Rodríguez F, Amato A, et al. 2023. Morphology and phylogeny of Prorocentrum porosum sp. nov. (Dinophyceae): a new benthic toxic dinoflagellate from the Atlantic and Pacific Oceans. Harmful Algae, 121: 102356, doi: 10.1016/j.hal.2022.102356

Blanco J, Moroño Á, Fernández M L. 2005. Toxic episodes in shellfish, produced by lipophilic phycotoxins: an overview. Galician: Revista Galega dos Recursos Mariños Galician Journal of Marine Resources, 1: 1–70

Boopathi T, Faria D G, Cheon J Y, et al. 2015. Implications of high molecular divergence of nuclear rRNA and phylogenetic structure for the dinoflagellate Prorocentrum (Dinophyceae, Prorocentrales). Journal of Eukaryotic Microbiology, 62(4): 519–531, doi: 10.1111/jeu.12206

Borsato G T, Salgueiro F, De’Carli G A L, et al. 2023. Taxonomy and abundance of epibenthic Prorocentrum (Dinophyceae) species from the tropical and subtropical Southwest Atlantic Ocean including a review of their global diversity and distribution. Harmful Algae, 127: 102470, doi: 10.1016/j.hal.2023.102470

Chomérat N, Bilien G, Zentz F. 2019. A taxonomical study of benthic Prorocentrum species (Prorocentrales, Dinophyceae) from Anse Dufour (Martinique Island, eastern Caribbean Sea). Marine Biodiversity, 49(3): 1299–1319, doi: 10.1007/s12526-018-0913-6

Chomérat N, Saburova M, Bilien G, et al. 2012. Prorocentrum bimaculatum sp. nov. (Dinophyceae, Prorocentrales), a new benthic dinoflagellate species from Kuwait (Arabian Gulf). Journal of Phycology, 48(1): 211–221, doi: 10.1111/j.1529-8817.2011.01102.x

Chomérat N, Sellos D Y, Zentz F, et al. 2010. Morphology and molecular phylogeny of Prorocentrum consutum sp. nov. (Dinophyceae), a new benthic dinoflagellate from south brittany (northwestern France). Journal of Phycology, 46(1): 183–194, doi: 10.1111/j.1529-8817.2009.00774.x

Chomérat N, Zentz F, Boulben S, et al. 2011. Prorocentrum glenanicum sp. nov. and Prorocentrum pseudopanamense sp. nov. (Prorocentrales, Dinophyceae), two new benthic dinoflagellate species from South Brittany (northwestern France). Phycologia, 50(2): 202–214, doi: 10.2216/10-12.1

Dodge J D. 1975. The Prorocentrales (Dinophyceae). II. Revision of the taxonomy within the genus Prorocentrum. Botanical Journal of the Linnean Society, 71(2): 103–125, doi: 10.1111/j.1095-8339.1975.tb02449.x

Ehrenberg C. 1834. Dritter beitrag zur erkenntniss grosser organisation in der richtung des kleinsten raumes. Berlin: Abhandlungen der Königlichen Akademie der Wissenschaften zu, 1833: 145–336

Ernst M, Kang K B, Caraballo-Rodríguez A M, et al. 2019. MolNetEnhancer: enhanced molecular networks by integrating metabolome mining and annotation tools. Metabolites, 9(7): 144, doi: 10.3390/metabo9070144

European Food Safety Authority. 2008. Marine biotoxins in shellfish‐okadaic acid and analogues‐scientific opinion of the panel on contaminants in the food chain. European Food Safety Authority Journal, 6(1): 589, doi: 10.2903/j.efsa.2008.589

Faust M A, Vandersea M W, Kibler S R, et al. 2008. Prorocentrum levis, a new benthic species (Dinophyceae) from a mangrove island, Twin Cays, Belize. Journal of Phycology, 44(1): 232–240, doi: 10.1111/j.1529-8817.2007.00450.x

Freudenthal A, Jijina J. 1988. Potential hazards of Dinophysis to consumers and shellfisheries. Journal of Shellfish Research, 7(1): 157–158

Fu X H, Meng F L, Hu Y, et al. 2008. Candida albicans, a distinctive fungal model for cellular aging study. Aging Cell, 7(5): 746–757, doi: 10.1111/j.1474-9726.2008.00424.x

Glibert P M, Burkholder J M, Kana T M. 2012. Recent insights about relationships between nutrient availability, forms, and stoichiometry, and the distribution, ecophysiology, and food web effects of pelagic and benthic Prorocentrum species. Harmful Algae, 14: 231–259, doi: 10.1016/j.hal.2011.10.023

Gómez F, Gourvil P, Li T C, et al. 2023. Molecular phylogeny of the spiny‐surfaced species of the dinoflagellate Prorocentrum with the description of P. thermophilum sp. nov. and P. criophilum sp. nov. (Prorocentrales, Dinophyceae). Journal of Phycology, 59(1): 70–86, doi: 10.1111/jpy.13298

Grzebyk D, Sako Y, Berland B. 1998. Phylogenetic analysis of nine species of Prorocentrum (Dinophyceae) inferred from 18S ribosomal DNA sequences, morphological comparisons, and description of Prorocentrum panamensis, sp. nov. Journal of Phycology, 34(6): 1055–1068, doi: 10.1046/j.1529-8817.1998.341055.x

Gul S, Saifullah S M. 2011. The dinoflagellate genus Prorocentrum (Prorocentrales, Prorocentraceae) from the north Arabian sea. Pakistan Journal of Botany, 43(6): 3061–3065

Gurevich A, Mikheenko A, Shlemov A, et al. 2018. Increased diversity of peptidic natural products revealed by modification-tolerant database search of mass spectra. Nature Microbiology, 3(3): 319–327, doi: 10.1038/s41564-017-0094-2

Hall T A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41: 95–98

Han M S, Wang Pengbin, Kim J H, et al. 2016. Morphological and molecular phylogenetic position of Prorocentrum micans sensu stricto and description of Prorocentrum koreanum sp. nov. from southern coastal waters in Korea and Japan. Protist, 167(1): 32–50, doi: 10.1016/j.protis.2015.12.001

Henrichs D W, Scott P S, Steidinger K A, et al. 2013. Morphology and phylogeny of Prorocentrum texanum sp. nov. (Dinophyceae): a new toxic dinoflagellate from the Gulf of Mexico coastal waters exhibiting two distinct morphologies. Journal of Phycology, 49(1): 143–155, doi: 10.1111/jpy.12030

Holman J D, Tabb D L, Mallick P. 2014. Employing ProteoWizard to convert raw mass spectrometry data. Current Protocols in Bioinformatics, 46(1): 13.24. 1–13.24. 9, doi: 10.1002/0471250953.bi1324s46

Hong D D, Thu N T H, Nam H S, et al. 2008. The phylogenetic tree of Alexandrium, Prorocentrum and Pseudo-nitzschia of harmful and toxic algae in vietnam coastal waters based on sequences of 18s rDNA, ITS1-5.8S-ITS2 gene fragments and single cell-PCR method. Marine Research in Indonesia, 32(2): 203–218, doi: 10.14203/mri.v32i2.456

Hoppenrath M. 2000. A new marine sand-dwelling Prorocentrum species, P. clipeus sp. nov. (Dinophyceae, Prorocentrales) from Helgoland, German Bight, North Sea. European Journal of Protistology, 36(1): 29–33, doi: 10.1016/S0932-4739(00)80019-X

Hoppenrath M, Chomérat N, Horiguchi T, et al. 2013. Taxonomy and phylogeny of the benthic Prorocentrum species (Dinophyceae)—a proposal and review. Harmful Algae, 27: 1–28, doi: 10.1016/j.hal.2013.03.006

Hoppenrath M, Leander B S. 2008. Morphology and molecular phylogeny of a new marine sand-dwelling Prorocentrum species, P. tsawwassenense (Dinophyceae, Prorocentrales), from British Columbia, Canada. Journal of Phycology, 44(2): 451–466, doi: 10.1111/j.1529-8817.2008.00483.x

Hoppenrath M, Murray S A, Chomérat N, et al. 2014. Marine Benthic Dinoflagellates-Unveiling Their Worldwide Biodiversity. Stuttgart: Senckenberg, 137–138

Keller M D, Selvin R C, Claus W, et al. 1987. Media for the culture of oceanic ultraphytoplankton. Journal of Phycology, 23(4): 633–638, doi: 10.1111/j.1529-8817.1987.tb04217.x

Jackson A E, Marr J C, Mclachlan J L. 1993. The production of diarrhetic shellfish toxins by an isolate of Prorocentrum lima from Nova Soctia, Canada. In: Smayda TJ, Shimizu Y, eds. Toxic Phytoplankton Blooms in the Sea. Newport: Elsevier, 513–518

Lim Z F, Luo Zhaohe, Lee L K, et al. 2019. Taxonomy and toxicity of Prorocentrum from Perhentian Islands (Malaysia), with a description of a non-toxigenic species Prorocentrum malayense sp. nov. (Dinophyceae). Harmful Algae, 83: 95–108, doi: 10.1016/j.hal.2019.01.007

Luo Zhaohe, Zhang Hua, Krock B, et al. 2017. Morphology, molecular phylogeny and okadaic acid production of epibenthic Prorocentrum (Dinophyceae) species from the northern South China Sea. Algal Research, 22: 14–30, doi: 10.1016/j.algal.2016.11.020

Mohimani H, Gurevich A, Shlemov A, et al. 2018. Dereplication of microbial metabolites through database search of mass spectra. Nature Communications, 9(1): 4035, doi: 10.1038/s41467-018-06082-8

Murray S. 2003. Diversity and phylogenetics of sand-dwelling dinoflagellates from southern Australia [dissertation]. Sydney: University of Sydney

Murray S, Nagahama Y, Fukuyo Y. 2007. Phylogenetic study of benthic, spine-bearing prorocentroids, including Prorocentrum fukuyoi sp. nov. Phycological Research, 55(2): 91–102, doi: 10.1111/j.1440-1835.2007.00452.x

Nascimento S M, Mendes M C Q, Menezes M, et al. 2017. Morphology and phylogeny of Prorocentrum caipirignum sp. nov. (Dinophyceae), a new tropical toxic benthic dinoflagellate. Harmful Algae, 70: 73–89, doi: 10.1016/j.hal.2017.11.001

Nothias L F, Petras D, Schmid R, et al. 2020. Feature-based molecular networking in the GNPS analysis environment. Nature Methods, 17(9): 905–908, doi: 10.1038/s41592-020-0933-6

Olmos-Soto J, Paniagua-Michel J, Contreras R, et al. 2002. Molecular identification of β-carotene hyper-producing strains of Dunaliella from saline environments using species-specific oligonucleotides. Biotechnology Letters, 24(5): 365–369, doi: 10.1023/A:1014516920887

Prabowo D A, Agusti S. 2019. Free-living dinoflagellates of the central Red Sea, Saudi Arabia: Variability, new records and potentially harmful species. Marine Pollution Bulletin, 141: 629–648, doi: 10.1016/j.marpolbul.2019.03.012

Rein K S, Snyder R V. 2006. The biosynthesis of polyketide metabolites by dinoflagellates. Advances in Applied Microbiology, 59: 93–125, doi: 10.1016/S0065-2164(06)59004-5

Ronquist F, Teslenko M, Van Der Mark P, et al. 2012. MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology, 61(3): 539–542, doi: 10.1093/sysbio/sys029

Schlitzer R. 2023. Ocean data view. http://odv.awi.de

Schmid R, Heuckeroth S, Korf A, et al. 2023. Integrative analysis of multimodal mass spectrometry data in MZmine 3. Nature Biotechnology, 41(4): 447–449, doi: 10.1038/s41587-023-01690-2

Scholin C A, Herzog M, Sogin M, et al. 1994. Identification of group‐and strain‐specific genetic markers for globally distributed Alexandrium (Dinophyceae). II. Sequence analysis of a fragment of the LSU rRNA gene. Journal of Phycology, 30(6): 999–1011, doi: 10.1111/j.0022-3646.1994.00999.x

Shah M M R, An S J, Lee J B. 2013. Presence of benthic dinoflagellates around coastal waters of Jeju Island including newly recorded species. Journal of Ecology and Environment, 36(4): 347–370, doi: 10.5141/ecoenv.2013.347

Shannon P, Markiel A, Ozier O, et al. 2003. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Research, 13(11): 2498–2504, doi: 10.1101/gr.1239303

Swofford D L, 2002. PAUP: phylogenetic analysis using parsimony, version 4.0 b10. Sinauer Associates, Sunderland, MA

Ten-Hage L, Turquet J, Quod J P, et al. 2000. Prorocentrum borbonicum sp. nov. (Dinophyceae), a new toxic benthic dinoflagellate from the southwestern Indian Ocean. Phycologia, 39(4): 296–301, doi: 10.2216/i0031-8884-39-4-296.1

Thompson J D, Higgins D G, Gibson T J. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic acids research, 22(22): 4673–4680, doi: 10.1093/nar/22.22.4673

Tillmann U, Gottschling M, Wietkamp S, et al. 2023a. Morphological and phylogenetic characterisation of Prorocentrum spinulentum, sp. nov. (Prorocentrales, Dinophyceae), a small spiny species from the North Atlantic. Microorganisms, 11(2): 271, doi: 10.3390/microorganisms11020271

Tillmann U, Hoppenrath M, Gottschling M. 2019. Reliable determination of Prorocentrum micans Ehrenb. (Prorocentrales, Dinophyceae) based on newly collected material from the type locality. European Journal of Phycology, 54(3): 417–431, doi: 10.1080/09670262.2019.1579925

Tillmann U, Wietkamp S, Gottschling M, et al. 2023b. Prorocentrum pervagatum sp. nov. (Prorocentrales, Dinophyceae): a new, small, planktonic species with a global distribution. Phycological Research, 71(1): 56–71, doi: 10.1111/pre.12502

Verma A, Kazandjian A, Sarowar C, et al. 2019. Morphology and phylogenetics of benthic Prorocentrum Species (Dinophyceae) from tropical northwestern Australia. Toxins, 11(10): 571, doi: 10.3390/toxins11100571

Wan Xiukun, Yao Ge, Liu Yanli, et al. 2019. Research progress in the biosynthetic mechanisms of marine polyether toxins. Marine Drugs, 17(10): 594, doi: 10.3390/md17100594

Wu Yixuan, Huang Shuning, Krock B, et al. 2022. Cryptic speciation of benthic Prorocentrum (Dinophyceae) species and their potential as ecological indicators. Journal of Sea Research, 190: 102304, doi: 10.1016/j.seares.2022.102304

White T J, Bruns T, Lee S, et al. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis M A, Gelfand D H, Sninsky J J, et al, eds. PCR Protocols: A Guide to Methods and Applications. New York: Academic Press, 315–322, doi: 10.1016/B978-0-12-372180-8.50042-1

Xie Hang, Zou Jian, Zheng Chengzhi, et al. 2022. Biodiversity and distribution of benthic dinoflagellates in tropical Zhongsha Islands, South China Sea. Journal of Oceanology and Limnology, 40(6): 2120–2145, doi: 10.1007/s00343-022-1322-z

Yasumoto T, Murata M, Oshima Y, et al. 1985. Diarrhetic shellfish toxins. Tetrahedron, 41(6): 1019–1025, doi: 10.1016/S0040-4020(01)96469-5

Zhang Hua, Li Yang, Cen Jingyi, et al. 2015. Morphotypes of Prorocentrum lima (Dinophyceae) from Hainan island, South China Sea: morphological and molecular characterization. Phycologia, 54(5): 503–516, doi: 10.2216/15-8.1

Zou Jian, Li Qun, Liu Hui, et al. 2022. Taxonomy and toxin profile of harmful benthic Prorocentrum (Dinophyceae) species from the Xisha Islands, South China Sea. Journal of Oceanology and Limnology, 40(3): 1171–1190, doi: 10.1007/s00343-021-1045-6

Zou Jian, Li Qun, Lu Songhui, et al. 2020. The first benthic harmful dinoflagellate bloom in China: morphology and toxicology of Prorocentrum concavum. Marine Pollution Bulletin, 158: 111313, doi: 10.1016/j.marpolbul.2020.111313

Relative Articles

Supplements(0)

Cited By

Proportional views