Morphological and molecular evidence for the recognition of Caloglossa fonticola sp. nov. (Delesseriaceae, Rhodophyta) from an underground spring in Guangxi, China

Caloglossa (Ceramiales, Rhodophyta) is a globally distributed euryhaline red macroalgal genus. However, only a few Caloglossa species have been documented in permanent freshwater habitats. Here, a new Caloglossa species, Caloglossa fonticola sp. nov., is described and illustrated from an underground spring in Bama County, Guangxi, China, based on morphological observations and molecular phylogenetic analysis. The new species is morphologically distinguishable from other species in the genus by a unique combination of character states, including thallus internodal blades linear, hardly or slightly constricted at nodes, adventitious branches and endogenous branches both absent, and unicellular rhizoids forming from clusters of wing cells arising from the first three axial cells of the main axis and lateral axis. Phylogenetic analysis based on rbcL and LSU rRNA DNA sequences indicated that Caloglossa fonticola was nested in a well-supported clade with Caloglossa bengalensis (Bootstrap supports for ML/posterior probabilities for BI: rbcL-98/1.00, LSU-100/1.00), with high sequence divergence between these two species (8.2–8.4% for rbcL and 3.2% for LSU), supporting its specific status. This is the first species of the genus Caloglossa reported in spring water habitats. The description of C. fonticola expands the known diversity of Caloglossa.

References

Chou, J.-Y., Wen, Y.-D. & Wang, W.-L. (2014) Morphological and molecular data confirm new records of three freshwater red algae, Batrachospermum macrosporum, Nemalionopsis tortuosa and Caloglossa leprieurii in Taiwan. Nova Hedwigia 98 (1–2): 233–246. https://doi.org/10.1127/0029-5035/2013/0151
Drobnik, J. (2007) Herbarium and knowledge about herbaria. PWN, Warszawa. 293 pp.
Fang, K.P., Nan, F.R., Feng, J., Lv, J.P., Liu, Q., Liu, X.D. & Xie, S.L. (2021) Virescentia guangxiensis (Batrachospermales, Rhodophyta): a new freshwater red algal species from South China. Journal of Oceanology and Limnology 39: 1538–1546.  https://doi.org/10.1007/s00343-020-0225-0
Felsenstein, J. (1981) Evolutionary trees from DNA sequences: a maximum likelihood approach. Journal of Molecular Evolution 17 (6): 368–376.  https://doi.org/10.1007/BF01734359
Freshwater, D.W., Miller, C.E., Frankovich, T.A. & Wynne, M.J. (2021) DNA sequence analyses reveal two new species of Caloglossa (Delesseriaceae, Rhodophyta) from the Skin of West Indian Manatees. Journal of Marine Science and Engineering 9 (163): 1–12.  https://doi.org/10.3390/jmse9020163
Freshwater, D.W. & Rueness, J. (1994) Phylogenetic relationships of some European Gelidium (Gelidiales, Rhodophyta) species, based on rbcL nucleotide sequence analysis. Phycologia 33 (3): 187–194.  https://doi.org/10.2216/i0031-8884-33-3-187.1
Guindon, S. & Gascuel, O. (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Systematic Biology 52 (5): 696–704.  https://doi.org/10.1080/10635150390235520
Guiry, M.D. & Guiry, G.M. (2021) AlgaeBase. World-wide electronic publication, National University of Ireland, Galway. Available from: http://www.algaebase.org (accessed 24 August 2021)
Hall, T.A. (1999) Bioedit: a user-friendly biological sequence alignment program for windows 95/98/NT. Nucleic Acids Symposium Series 41 (41): 95–98.  https://doi.org/10.1021/bk-1999-0734.ch008
Harper, J.T. & Saunders, G.W. (2001) The application of sequences of the ribosomal cistron to the systematics and classification of the Florideophyte red algae (Florideophyceae, Rhodophyta). Cahiers De Biologie Marine 42: 25–38.  https://doi.org/10.1515/BOT.2001.012
Jao, C.-C. (1941) Studies on the freshwater algae of China. VIII. A preliminary account of the Chinese freshwater Rhodophyceae. Sinesia 12: 245–289.
Kalmbach, K. (2011) Herbarium plant collection protocol. How to prepare herbarium specimens? Denver Botanic Gardens, Denver, USA, 12 pp.
Kamiya, M., Tanaka, J., King, R.J., West, J.A., Zuccarello, G.C. & Kawai, H. (1999) Reproductive and genetic distinction between broad and narrow entities of Caloglossa continua (Delesseriaceae, Rhodophyta). Phycologia 38 (5): 356–367.  https://doi.org/10.2216/i0031-8884-38-5-356.1
Kamiya, M. & West, J.A. (2014) Cryptic diversity in the euryhaline red alga Caloglossa ogasawaraensis (Delesseriaceae, Ceramiales). Phycologia 53 (4): 374–382.  https://doi.org/10.2216/13-242.1
Kamiya, M., West, J.A., Karsten, U. & Ganesan, E.K. (2016) Molecular and morphological delineation of Caloglossa beccarii and related species (Delesseriaceae, Rhodophyta). Phycologia 55 (6): 640–649.  https://doi.org/10.2216/16-19.1
Kamiya, M., West, J.A. & Zuccarello, G.C. (2018) High species richness and genetic diversity of the genus Caloglossa (Delesseriaceae, Rhodophyta) in New Ireland, Papua New Guinea. Phycological Research 66 (3): 218–230.  https://doi.org/10.1111/pre.12223
Kamiya, M., Zuccarello, G.C. & West, J.A. (2003) Evolutionary relationships of the genus Caloglossa (Delesseriaceae, Rhodophyta) inferred from large-subunit ribosomal RNA gene sequences, morphological evidence and reproductive compatibility, with description of a new species from Guatemala. Phycologia 42 (5): 478–497.  https://doi.org/10.2216/i0031-8884-42-5-478.1
King, R. & Puttock, C. (1994) Morphology and taxonomy of Caloglossa (Delesseriaceae, Rhodophyta). Australian Systematic Botany 7: 89–124.  https://doi.org/10.1071/sb9940089
Krayesky, D.M., Norris, J.N., West, J.A. & Fredericq, S. (2011) The Caloglossa leprieurii complex (Delesseriaceae, Rhodophyta) in the Americas: the elucidation of overlooked species based on molecular and morphological evidence. Cryptogamie Algologie 31 (1): 37–62.  https://doi.org/10.7872/crya.v32.iss1.2011.037
Krayesky, D.M., Norris, J.N., West, J.A., Kamiya, M., Viguerie, M., Wysor, B.S. & Fredericq, S. (2012) Two new species of Caloglossa (Delesseriaceae, Rhodophyta) from the Americas, C. confusa and C. fluviatilis spp. nov. Phycologia 51 (5): 513–530.  https://doi.org/10.2216/11-57.1
Kumano, S. (2002) Freshwater Red Algae of the World. Biopress Ltd, Bristol, UK. 375 pp.
Papenfuss, G.F. (1961) The Structure and Reproduction of Caloglossa leprieurii. Phycologia 1 (1): 8–31.  https://doi.org/10.2216/i0031-8884-1-1-8.1
Posada, D. & Buckley, T.R. (2004) Model selection and model averaging in phylogenetics: advantages of Akaike information criterion and Bayesian approaches over likelihood ratio tests. Systematic Biology 53 (5): 793–808.  https://doi.org/10.1080/10635150490522304
Post, E. (1936) Systematische und pflanzengeographische Notizen zur Bostrychia-Caloglossa-Association. Revue Algologique 9: 1–84.
Ronquist, F. & Huelsenbeck, J.P. (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19 (12): 1572–1574.  https://doi.org/10.1093/bioinformatics/btg180
Rybak, A.S. (2018) The Ulva flexuosa complex (Ulvaceae, Chlorophyta): An updated identification key with special reference to the freshwater and hyperhaline taxa. Phytotaxa 345 (2): 83–103. https://doi.org/10.11646/phytotaxa.345.2.1
Sheath, R.G., Vis, M.L. & Cole, K.M. (1993) Distribution and systematics of freshwater Ceramiales (Rhodophyta) in North America. Journal of Phycology 29 (1): 108–117.  https://doi.org/10.1111/j.1529-8817.1993.tb00288.x
Shi, Z.-X. 2006. Flora algarum sinicarum aquae dulcis, Tomus Xiii, Rhodophyta, Phaeophyta. Beijing Science Press, Beijing, 208 pp.
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution 28 (10): 2731–2739.  http://dx.doi.org/10.1093/molbev/msr121
Thiers, B. (2021) (continuously updated). Index Herbariorum: A Global Directory of Public Herbaria and Associated Staff. New York Botanical Garden’s Virtual Herbarium, New York. Available from: http://sweetgum.nybg.org/ih/ (accessed 24 August 2021)
Tseng, C.K. (2001) Flora algarum marinarum sinicarum, Tomus II, Rhodophyta. No. VI Ceramiales. Beijing Science Press, Beijing, 159 pp.
Tseng, C.K. & Chang, C.F. (1962) Notes on Caloglossa, Digenea and the other anthelmintic marine algae in china. Acta Pharmaceutica sinaca 9 (3): 180–186.
Vis, M.L. & Sheath, R.G. (1997) Biogeography of Batrachospermum gelatinosum (Batrachospermales, Rhodophyta) in North America based on molecular and morphological data. Journal of Phycology 33 (3): 520–526.  https://doi.org/10.1111/j.0022-3646.1997.00520.x
West, J.A., Kamiya, M., Ganesan, E.K., Goer, L.D. & Jose, L. (2015) Caloglossa beccarii (Delesseriaceae, Rhodophyta) from freshwater rivers in Kerala, India, a critical new record. Algae 30 (3): 207–216.  https://doi.org/10.4490/algae.2015.30.3.207
West, J.A., Zuccarello, G.C. & Kamiya, M. (2001) Reproductive patterns of Caloglossa species (Delesseriaceae, Rhodophyta) from Australia and New Zealand: multiple origins of asexuality in C. leprieurii. literature review on apomixis, mixed-phase, bisexuality and sexual compatibility. Phycological Research 49 (3): 183–200.  https://doi.org/10.1046/j.1440-1835.2001.00239.x
West, J.A., Zuccarello, G.C., Pedroche, F.F. & Karsten, U. (1994) Caloglossa apomeiotica sp. nov. (Ceramiales, Rhodophyta) frorn Pacific México. Botanica Marina 37: 381–390.  https://doi.org/10.1515/botm.1994.37.4.381