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