Cyst-Motile Stage Relationship and Phylogenetic Study of Levanderina fissa, a Bloom-Forming Dinoflagellate from Chabahar Bay in the Coasts of the Sea of Oman

Document Type : Original Article

Authors

Department of Marine Biology, Faculty of Marine Sciences, Chabahar Maritime University, Chabahar, Iran

10.48308/pae.2026.244187.1141

Abstract

Levanderina fissa is an unarmored, cosmopolitan marine dinoflagellate recognized for its capacity to form harmful algal blooms (HABs), which can negatively impact marine ecosystems and coastal economies. Despite its ecological significance, knowledge of its life cycle, including the role of benthic cysts in bloom initiation and termination, remains limited particularly in the Sea of Oman. This study presents the first isolation and characterization of L. fissa cysts from sediments collected using a Van Veen grab in the Konarak region, along the southeastern coasts of Iran. Single cysts were isolated and germinated under controlled laboratory conditions; 12:12 light-dark photoperiod, 25°C, 60 µmol photons m⁻² s⁻¹ light illumination in f/2 medium. Morphological examination of the cysts revealed a spherical to ovoidal shape with a thick, unornamented wall. Following germination, the morphology of the resulting motile cells was examined using light microscopy, revealing key diagnostic features, including an unusual and distinctly formed sulcus, consistent with the description of L. fissa. For molecular confirmation and phylogenetic placement, the Internal Transcribed Spacer (ITS) region was amplified and sequenced. Phylogenetic analysis using Maximum Likelihood (ML) placed the Iranian strain firmly within a monophyletic clade containing other L. fissa strains, supported by a 98–100% bootstrap value. This combined morphological and molecular approach unequivocally confirms the species identity. This research underscores the critical importance of integrating cyst studies into HAB monitoring programs, as understanding the distribution and germination dynamics of benthic resting stages is essential for predicting bloom timing, magnitude, and spatial extent in vulnerable coastal ecosystems such as the Sea of
Levanderina fissa is a cosmopolitan, unarmored marine dinoflagellate capable of forming harmful algal blooms (HABs), which adversely impact marine ecosystems and coastal economies. Despite its ecological significance, knowledge of its life cycle, particularly the role of benthic cysts in bloom dynamics, remains limited, particularly in the Sea of Oman. This study presents the first isolation and characterization of L. fissa resting cysts from sediments collected using a Van Veen grab sampler in the Konarak region along the southeastern coasts of Iran. Single cysts were isolated and germinated in the f/2 medium at 25°C under a 12:12 light photoperiod with an irradiance of 60 µmol photons m⁻² s⁻¹. Morphological examination of the cysts revealed a spherical to ovoidal shape with a thick, unornamented wall. Following germination, the morphology of the resulting motile cells was examined using light microscopy, revealing key diagnostic features, including an unusual and distinctly formed sulcus, consistent with the description of L. fissa. For molecular confirmation and phylogenetic placement, the Internal Transcribed Spacer (ITS) region was amplified and sequenced. Phylogenetic analysis using Maximum Likelihood (ML) placed the Iranian strain firmly within a monophyletic clade containing other L. fissa strains, supported by a 98–100% bootstrap value. This combined morphological and molecular approach unequivocally confirms the species identity. This research underscores the critical importance of integrating cyst studies into HAB monitoring programs, as understanding the distribution and germination dynamics of benthic resting stages is essential for predicting bloom timing, magnitude, and spatial extent in vulnerable coastal ecosystems such as the Sea of Oman.
Oman.

Keywords

References

Adachi, M., Sake, Y., and Ishida, Y., 1996. Analysis of Alexandrium (Dinophyceae) species using sequences of the 5.8 S ribosomal DNA and internal transcribed spacer regions. Journal of Phycology, 32(3), pp.424–432. https://doi.org/10.1111/j.0022-3646.1996.00424.x.
Alpermann, T.J., Tillmann, U., Beszteri, B., Cembella, A.D. and John, U., 2010. Phenotypic variation and genotypic diversity in a planktonic population of the toxigenic marine dinoflagellate Alexandrium tamarense (Dinophyceae). Journal of Phycology, 46(1), pp.18–32. https://doi.org/10.1111/j.1529-8817.2009.00767.
Anjaneyan, P., Kuttippurath, J., Kumar, P.H., Ali, S.M., and Raman, M., 2023. Spatio-temporal changes of winter and spring phytoplankton blooms in Arabian sea during the period 1997–2020. Journal of Environmental Management, 332, p.117435. https://doi.org/10.1016/j.jenvman.2023.117435.
Attaran Fariman, G. and Bolch, C.J.S., 2014. Morphology and genetic affinities of a novel Chattonella isolate (Raphidophyceae) isolated from Iran’s south coast (Oman Sea). Turkish Journal of Botany, 38(1), pp.156–168. https://doi.org/10.3906/bot-1210-33.
Attaran Fariman, G., Khalili, M., and Sadeghi, P., 2018a. Effects of environmental parameters on dinoflagellates cysts of Gonyaulacales and Gymnodiales in the Oman Sea coasts and the Strait of Hormuz. ISFJ, 26(6), pp.13–23. https://doi.org/10.22092/ISFJ.2018.115696.
Attaran Fariman, G., Khodami, S. and Bolch, C.J.S., 2011. The cyst-motile stage relationship of three Protoperidinium species from south-east coast of Iran. Iranian Journal of Fisheries Sciences, 10(1), pp.1–12.
Attaran Fariman, G., Loghmani, M., and Mirkazehi, R.A., 2018b. The effect of environmental factors on the abundance of dinoflagellate cysts in sediments of the Southeastern Coast of Iran in hot and cold seasons. Journal of Oceanography, 9(34), pp.59–67. https://sid.ir/paper/158132/en.
Attaran-Fariman, G. and Bolch, C.J., 2007. Scrippsiella irregularis sp. nov. (Dinophyceae), a new dinoflagellate from the southeast coast of Iran. Phycologia, 46(5), pp.572–582. https://doi.org/10.2216/07-02.1.
Attaran-Fariman, G., and Bolch, C.J.S., 2012. Morphology and phylogeny of Scrippsiella trochoidea (Dinophyceae) a potentially harmful bloom forming species isolated from the sediments of Iran’s south coast. Iranian Journal of Fisheries Sciences, 11(2), pp.252–270.
Attaran-Fariman, G., Abir, S., Dolatabadi, F., Abedi, E. and Sayareh, F., 2025. Dinoflagellate cyst assemblages in recent sediments of Iran’s tropical mangrove forests. Marine Micropaleontology, 195, p.102449. https://doi.org/10.1016/j.marmicro.2025.102449.
Attaran-Fariman, G., De Salas, M.F., Negri, A.P. and Bolch, C.J., 2007. Morphology and phylogeny of Gymnodinium trapeziforme sp. nov. (Dinophyceae): a new dinoflagellate from the southeast coast of Iran that forms microreticulate resting cysts. Phycologia, 46(6), pp.644–656. https://doi.org/10.2216/07-05.1.
Attaran-Fariman, G., Hashemzai, M. and Khajeh Amiri Khaledi, C., 2017. Investigation of diversity and distribution of phytoplankton cysts in the recent sediments of the western Chabahar coasts. Journal of Oceanography, 8(29), pp.65–74. https://doi.org/10.18869/acadpub.joc.8.29.65.
Attaran-Fariman, G., Khodami, S. and Bolch, C.J., 2012. First observation of dinoflagellate resting cysts from recent sediments of the southeast coast of Iran. Algological Studies, 140(1), pp.51–79. https://doi.org/10.1127/1864-1318/2012/0048.
Baula, I.U., Azanza, R.V., Fukuyo, Y. and Siringan, F.P., 2011. Dinoflagellate cyst composition, abundance and horizontal distribution in Bolinao, Pangasinan, Northern Philippines. Harmful Algae, 11, pp.33–44. https://doi.org/10.1016/j.hal.2011.07.002.
Bolch, C.J. and Hallegraeff, G.M., 1990. Dinoflagellate cysts in recent marine sediments from Tasmania, Australia. Botanica Marina, 33(2). https://doi.org/10.1515/botm.1990.33.2.173.
Borbor-Cordova, M.J., Torres, G., Mantilla-Saltos, G., Casierra-Tomala, A., Bermúdez, J.R., Renteria, W. and Bayot, B., 2019. Oceanography of harmful algal blooms on the Ecuadorian coast (1997–2017): integrating remote sensing and biological data. Frontiers in Marine Science, 6, p.13. https://doi.org/10.3389/fmars.2019.00013.
Bravo, I. and Figueroa, R.I., 2014. Towards an ecological understanding of dinoflagellate cyst functions. Microorganisms, 2(1), pp.11–32. https://doi.org/10.3390/microorganisms2010011.
Caroppo, C., Denti, G. and Rubino, F., 2025. Active and resting harmful dinoflagellates in the highly productive mussel culture area of the Mar Piccolo (northern Ionian Sea). Biologia Marina Mediterranea, 29(1), pp.146–149. https://doi.org/10.82008/bmm.v29i1.321.
Chen, H., Yin, X., Chen, Y., Wang, Y., Li, Q., Ji, N., Zhou, L., Hu, G. and Shen, X., 2024. Characterization of a Levanderina fissa bloom in aquaculture ponds and its utilization of dissolved organic phosphorus. Microorganisms, 12(11), p.2202. https://doi.org/10.3390/microorganisms12112202.
Coats, D.W. and Park, M.G., 2002. Parasitism of photosynthetic dinoflagellates by three strains of Amoebophrya (Dinophyta): parasite survival, infectivity, generation time, and host specificity. Journal of Phycology, 38(3), pp.520–528. https://doi.org/10.1046/j.1529-8817.2002.01200.x
Dale, B., 1983. Dinoflagellate resting cysts: “benthic plankton”. In G.A. Fryxell (Ed.), Survival Strategies of the Algae (pp. 69–136). Cambridge University Press.
Dolatabadi, F., Attaran-Fariman, G. and Loghmani, M., 2021. Bloom occurrence and phylogeny of Gonyaulax polygramma (Dinophyceae) isolated from south east coast of Iran (Oman Sea). Iranian Journal of Fisheries Sciences, 20(6), pp.1789–1803. https://doi.org/10.22092/ijfs.2021.125501.
Ershadifar, H., Koochaknejad, E., Ghazilou, A., Kor, K., Negarestan, H. and Baskaleh, G., 2020. Response of phytoplankton assemblages to variations in environmental parameters in a subtropical bay (Chabahar Bay, Iran): harmful algal blooms and coastal hypoxia. Regional Studies in Marine Science, 39, p.101421. https://doi.org/10.1016/j.rsma.2020.101421.
Gárate-Lizárraga, I., 2014. Proliferation of Levanderina fissa and Polykrikos hartmannii (Dinophyceae: Gymnodiniales) in Bahía De La Paz, Gulf of California, México. CICIMAR Oceánides, 29(2), pp.25–35. https://doi.org/10.37543/oceanides.v29i2.137.
Gu, H., Luo, Z., Liu, T. and Lan, D., 2013. Morphology and phylogeny of Scrippsiella enormis sp. nov. and S. cf. spinifera (Peridiniales, Dinophyceae) from the China Sea. Phycologia, 52(2), pp.182–190. https://doi.org/10.2216/12-036.1.
Gu, H., Luo, Z., Mertens, K.N., Price, A.M., Turner, R.E. and Rabalais, N.N., 2015. Cyst-motile stage relationship, morphology, ultrastructure, and molecular phylogeny of the gymnodinioid dinoflagellate Barrufeta resplendens comb. nov., formerly known as Gyrodinium resplendens, isolated from the Gulf of Mexico. Journal of Phycology, 51(5), pp.990–999. https://doi.org/10.1111/jpy.12342.
Gu, H., Sun, J., Kooistra, W.H. and Zeng, R., 2008. Phylogenetic position and morphology of thecae and cysts of Scrippsiella (Dinophyceae) species in the East China Sea. Journal of Phycology, 44(2), pp.478–494. https://doi.org/10.1111/j.1529-8817.2008.00478.x.
Guillard, R.R. and Ryther, J.H., 1962. Studies of marine planktonic diatoms: I. Cyclotella nana Hustedt, and Detonula confervacea (Cleve) Gran. Canadian Journal of Microbiology, 8(2), pp.229–239. https://doi.org/10.1139/m62-029.
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, pp.95–98.
Hallegraeff, G.M., Anderson, D.M., Cembella, A.D. and Enevoldsen, H.O. (Eds.), 2004. Manual on Harmful Marine Microalgae (2nd ed., Monographs on Oceanographic Methodology, Vol. 11). UNESCO Publishing. http://unesdoc.unesco.org/images/0013/001317/131711e.
Huang, C. and Dong, C., 2000. Provisional names of some dinoflagellates from Chinese coastal waters. Chinese Journal of Oceanology and Limnology, 18(3), pp.278–285.
John, U., Litaker, R.W., Montresor, M., Murray, S., Brosnahan, M.L. and Anderson, D.M., 2014. Formal revision of the Alexandrium tamarense species complex (Dinophyceae) taxonomy: the introduction of five species with emphasis on molecular-based (rDNA) classification. Protist, 165(6), pp.779–804. https://doi.org/10.1016/j.protis.2014.10.001.
Kim, K.Y. and Kim, C.H., 2007. Phylogenetic relationships among diverse dinoflagellate species occurring in coastal waters off Korea inferred from large subunit ribosomal DNA sequence data. Algae, 22(2), pp.57–67. https://www.researchgate.net/publication/264150365.
Kojima, N. and Kobayashi, S., 1992. Motile cell-like cyst of Gyrodinium instriatum Freudenthal et Lee (Dinophyceae). Review of Palaeobotany and Palynology, 74(3–4), pp.239–247. https://doi.org/10.1016/0034-6667(92)90009-6.
Kremp, A. and Parrow, M.W., 2006. Evidence for asexual resting cysts in the life cycle of the marine peridinoid dinoflagellate, Scrippsiella hangoei. Journal of Phycology, 42(2), pp.400–409. https://doi.org/10.1111/j.1529-8817.2006.00205.x.
Lewis, J. and Hallett, R., 1997. Lingulodinium polyedrum (Gonyaulax polyedra): a blooming dinoflagellate. In A.D. Ansell, R.N. Gibson and M. Barnes (Eds.), Oceanography and Marine Biology: An Annual Review, Vol. 35, pp. 97–161. UCL Press.
Liu, D., Shi, Y., Di, B., Sun, Q., Wang, Y., Dong, Z. and Shao, H., 2012. The impact of different pollution sources on modern dinoflagellate cysts in Sishili Bay, Yellow Sea, China. Marine Micropaleontology, 84–85, pp.1–13. https://doi.org/10.1016/j.marmicro.2011.11.001.
Marret, F. and Zonneveld, K.A., 2003. Atlas of modern organic-walled dinoflagellate cyst distribution. Review of Palaeobotany and Palynology, 125(1–2), pp.1–200. https://doi.org/10.1016/S0034-6667(02)00229-4.
Matsuoka, K., 1999. Eutrophication process recorded in dinoflagellate cyst assemblages — a case of Yokohama Port, Tokyo Bay, Japan. Science of the Total Environment, 231(1), pp.17–35. https://doi.org/10.1016/S0048-9697(99)00087-X.
Matsuoka, K., Kawami, H., Fujii, R. and Iwataki, M., 2006. Further examination of the cyst-theca relationship of Protoperidinium thulesense (Peridiniales, Dinophyceae) and the phylogenetic significance of round brown cysts. Phycologia, 45(6), pp.632–641. https://doi.org/10.2216/05-42.1.
Mertens, K.N., Aydin, H., Uzar, S., Takano, Y., Yamaguchi, A. and Matsuoka, K., 2015. Relationship between the dinoflagellate cyst Spiniferites pachydermus and Gonyaulax ellegaardiae sp. nov. from Izmir Bay, Turkey, and molecular characterization. Journal of Phycology, 51(3), pp.560–573. https://doi.org/10.1111/jpy.12304.
Mertens, K.N., Yamaguchi, A., Takano, Y., Pospelova, V., Head, M.J., Radi, T., Pieńkowski, A.J., De Vernal, A., Kawami, H. and Matsuoka, K., 2013. A new heterotrophic dinoflagellate from the North-eastern Pacific, Protoperidinium fukuyoi: cyst-theca relationship, phylogeny, distribution and ecology. Journal of Eukaryotic Microbiology, 60(6), pp.545–563. https://doi.org/10.1111/jeu.12058.
Moestrup, Ø., Hakanen, P., Hansen, G., Daugbjerg, N. and Ellegaard, M., 2014. On Levanderina fissa gen. & comb. nov. (Dinophyceae) (syn. Gymnodinium fissum, Gyrodinium instriatum, Gyr. uncatenum), a dinoflagellate with a very unusual sulcus. Phycologia, 53(3), pp.265–292. https://doi.org/10.2216/13-254.1.
Nagasoe, S., Kim, D.I., Shimasaki, Y., Oshima, Y., Yamaguchi, M. and Honjo, T., 2006. Effects of temperature, salinity and irradiance on the growth of the red tide dinoflagellate Gyrodinium instriatum Freudenthal et Lee. Harmful Algae, 5(1), pp.20–25. https://doi.org/10.1016/j.hal.2005.06.001.
Orlova, T.Y., Selina, M.S. and Shevchenko, O.G., 2003. The morphology of cysts and motile cells of Gyrodinium instriatum (Dinophyta), a species new to the seas of Russia. Russian Journal of Marine Biology, 29(2), pp.120–122. https://doi.org/10.1023/A:1024037625075.
Partensky, F., Vaulot, D., Couté, A. and Sournia, A., 1988. Morphological and nuclear analysis of the bloom-forming dinoflagellates Gyrodinium cf. aureolum and Gymnodinium nagasakiense. Journal of Phycology, 24(3), pp.408–415. https://doi.org/10.1111/j.1529-8817.1988.tb04243.x.
Raisi, A., Attaran, G., Zare, R., and Asefi, M.A., 2023. Dinoflagellate cyst distribution in recent sediments of Gwater Bay in the northeast of Oman Sea in the pre- and post-monsoon season. Plant, Algae, and Environment, 7(1), pp.1003–1015. https://doi.org/10.48308/jpr.2023.232134.1048.
Ribeiro, S., Lundholm, N., Amorim, A. and Ellegaard, M., 2010. Protoperidinium minutum (Dinophyceae) from Portugal: cyst-theca relationship and phylogenetic position on the basis of single-cell SSU and LSU rDNA sequencing. Phycologia, 49(1), pp.48–63. https://doi.org/10.2216/09-11.1.
Shao, P., Chen, Y.Q., Zhou, H., Yuan, J., Qu, L.H., Zhao, D. and Lin, Y.S., 2004. Genetic variability in Gymnodiniaceae ITS regions: implications for species identification and phylogenetic analysis. Marine Biology, 144(2), pp.215–224. https://www.researchgate.net/publication/226014015.
Tamura, K., Stecher, G., Peterson, D., Filipski, A., and Kumar, S., 2013. MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution, 30(12), pp.2725–2729. https://doi.org/10.1093/molbev/mst197.
Thompson, J.D., Higgins, D.G. and 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), pp.4673–4680. https://doi.org/10.1093/nar/22.22.4673.
Yedema, Y.W., Donders, T., Peterse, F. and Sangiorgi, F., 2023. Dinoflagellate cyst and pollen assemblages as tracers for marine productivity and river input in the northern Gulf of Mexico. Journal of Micropalaeontology, 42(2), pp.257–276. https://doi.org/10.5194/jm-42-257-2023.
Plant, Algae, and Environment
Volume 10, Issue 2
DOi
2026
Pages 42-57

Files

Share

How to cite

Statistics