Bostrychia australiana J.A.West

ZuccareLLo School, Giuseppe C., 2023, Bostrychia australiana from Australia, a new species segregated from B. kelanensis Grunow ex Post (Rhodomelaceae, Rhodophyta), Notulae Algarum 268 : -

Bostrychia australiana J.A.West , Loiseaux-de Goër & ZuccareLLo, sp. nov.

Description: ThaLLus of prostrate and suberect indeterminate axes, sLender, turf forming; attachment often stipitate; axes 25 mm Long, unbranched or with one order of determinate LateraLs; 3-4 tiers of pericentraL ceLLs per axiaL ceLL, 5-6 pericentraL ceLLs around the indeterminate axes, essentiaLLy ecorticate throughout, with the exception of irreguLar cortication of the stipe; cLadohaptera not seen; carpogoniaL branches borne within the apicaL or intercaLary axiaL ceLLs; Cystocarps smaLL, with few spores in cuLture. SpermatangiaL stichidia as typicaL of the genus.

HoLotype: Cossack , Western AustraLia, 24°53'S. 117°11'E, 9 xii 1997, on a mangrove ( Avicennia sp. ), dried specimen from cuLture, JAW 3810 (femaLe gametophyte), UC 2050473 . HoLotype (JAW3810)

Accession Numbers: rbc L: OQ291113; COI: OQ291130; Rubisco Spacer: OQ297673.

Paratype: Mandorah , Northern Territory, AustraLia, 12°26'S 130°46'E, 21 vi 1997, on a mangrove ( Rhizophora sp. ), dried specimen from cuLture JAW 3748, UC 2050596 .

PhycoBank Registration: 103599.

Crosses attempted with a shaker (70 rev min-1) for 4 months of a seLf-cross (JAW 3810 maLe x JAW 3810 femaLe) showed spermatia on about 10% of trichogynes, but no pseudocystocarps (i.e. enLarged pericarp but no sporangiaL deveLopment) were produced and onLy one cystocarp ( Fig. 5 F) deveLoped. This cystocarp deveLoped about 10 carpospores that reLeased but did not germinate. A cross between two different isoLates (JAW 3810 maLe x JAW 3348 femaLe) produced over 10 pseudocystocarps but no viabLe cystocarp. A cross between a JAW 3810 maLe and a JAW 4261 femaLe produced no pseudocystocarps or cystocarps.

Reproduction in cuLture: Reproduction in cuLture was variabLe in B. kelanensis . Some isoLates produced no viabLe tetraspores (JAW 3075, 3076); IsoLate JAW 3349 reLeased tetraspores but

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sporeLings did not deveLop; Some isoLates reLeased tetraspores and produced maLe and femaLe gametophytes (JAW 3000, 3860, 4589, 4632, 4635), maLes often died, but occasionaLLy a surviving femaLe switched to maLe; Some reLeased tetraspores and onLy produced femaLe gametophytes (JAW 3213, 3214, 3348, 4261). B. australiana : JAW 3810 aLso reLeased tetraspores that produced maLe and femaLe gametophytes. JAW 3748 was a femaLe (derived from a tetrasporophyte) that was rareLy bisexuaL, but carposporophytes were never produced. This unusuaL reproduction in cuLture is found in some other isoLates of Bostrychia . For exampLe, Bostrychia tenella isoLates have mixed-phase patterns (see West & CaLumpong 1988) and some B. moritziana isoLates recycLe tetrasporophytes (see West & aL. 1992, West & ZuccareLLo 1999). Bostrychia radicosa recycLes tetrasporophytes and has mixed-phase reproduction, either maLes or femaLes on tetrasporophytes (see West & aL. 2006). Many other isoLates of Bostrychia have reguLar Polysiphonia - type sexuaL cycLes. What Leads to these unusuaL patterns and how they are manifested in the fieLd, if they are, needs further investigation.

Note: WhiLe cLadohaptera are said to be Less common in Bostrychia kelanensis (e.g. King & Puttock 1989), none were observed in fieLd or cuLture specimens of this species, and basaL discs were the onLy attachment structures seen. CLadohaptera was aLso Lacking in B. australiana .

In concLusion, Bostrychia australiana sp. nov. had a different and Less variabLe pattern of LongitudinaL pericentraL ceLL deveLopment (3-4 LongitudinaL pericentraL ceLLs per axiaL ceLL) compared to B. kelanensis (3-5 LongitudinaL pericentraL ceLLs per axiaL ceLL). CLadohaptera are absent in our fieLd and cuLture specimens, even though they are stated to be a key character of B. kelanensis by King & Puttock (1989). WhiLe B. australiana shows Less variabiLity than B. kelanensis , the morphoLogicaL characters of the two species overLap and B. australiana can be considered a true cryptic species. The naming of cryptic species is important for cataLoguing diversity in various areas. WhiLe this is easiLy tractabLe in cryptic species compLexes with onLy a few species, as in “ B. kelanensis ”, this may be more onerous in species compLexes with many cryptic species (Payo & aL. 2013, Vieira & aL. 2014, Muangmai & aL. 2022). The probLem of naming is exacerbated when cryptic species overLap in distribution: B. kelanensis and B. australiana are found in cLose proximity around Darwin (Northern Territory). OnLy sequencing of types wiLL cLarify the correct name assignment and Lead to naming the new species (Hughey & aL. 2001). To date, B. australiana has been found onLy in AustraLia, but further sampLing (e.g. neighbouring Indonesian isLands) may reveaL it to be more widespread, aLthough we did not find it in Papua New Guinea (unpubL. data).

DeLić, T., TronteLj, P., Rendoš, M. & Fišer, C. (2017). The importance of naming cryptic species and the conservation of endemic subterranean amphipods. Scientific Reports 7: 1–12.

Fraser, C. I., ZuccareLLo, G. C., Spencer, H. G., SaLvatore, L. C., Garcia, G. R. & Waters, J. M. (2013). Genetic affinities between trans-oceanic popuLations of non-buoyant macroaLgae in the high Latitudes of the southern hemisphere. PLoS ONE 8: e 69138.

Freshwater, D. W. & Rueness, J. (1994). PhyLogenetic reLationships of some European Gelidium (GeLidiaLes, Rhodophyta) species, based on rbc L nucLeotide sequence anaLysis. Phycologia 33: 187–194.

Ganesan, E.K., West, J.A. & Necchi, O., Jr. (2018). A cataLogue and bibLiography of non-marine (freshwater and estuarine) Rhodophyta (red aLgae) of India. Phytotaxa 364: 1–48.

Guiry, M.D. & Guiry, G.M. 2022. ALgaeBase. WorLd-wide eLectronic pubLication, University of GaLway. https://www.aLgaebase.org; searched on 18 December 2022.

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: 356–67.

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Karsten, U., West, J. A., ZuccareLLo, G. C. & Kirst, G. (1994). PhysioLogicaL ecotypes in the marine red aLga Bostrychia radicans (CeramiaLes, Rhodophyta) from the east coast of the USA. Journal of Phycology 30: 174-182.

King, R. J. & Puttock, C. F. (1989). MorphoLogy and taxonomy of Bostrychia and Stictosiphonia (RhodomeLaceae/Rhodophyta). Australian Systematic Botany 2: 1-73.

Minh, B. Q., Schmidt, H. A., Chernomor, O., Schrempf, D., Woodhams, M. D., HaeseLer, A. von & Lanfear, R. (2020). IQ-TREE 2: New modeLs and efficient methods for phyLogenetic inference in the genomic era. Molecular Biology and Evolution 37: 1530–1534.

Muangmai, N., West, J. A. & ZuccareLLo, G. C. (2014). EvoLution of four Southern Hemisphere B ostrychia (RhodomeLaceae, Rhodophyta) species: phyLogeny, species deLimitation and divergence times. Phycologia 53: 593–601.

Muangmai, N., Preuss, M. & ZuccareLLo, G. C. (2015 a). Comparative physioLogicaL studies on the growth of cryptic species of Bostrychia intricata (RhodomeLaceae, Rhodophyta) in various saLinity and temperature conditions. Phycological Research. 63: 300–306.

Muangmai, N., Fraser, C. & ZuccareLLo, G. C. (2015 b). Contrasting patterns of popuLation structure and demographic history in cryptic species of Bostrychia intricata (RhodomeLaceae, Rhodophyta) from New ZeaLand. Journal of Phycology 51: 574–585.

Muangmai, N., Ammon, U. von, & ZuccareLLo, G. C. (2016). Cryptic species in sympatry: nonrandom smaLL-scaLe distribution patterns in Bostrychia intricata (CeramiaLes, Rhodophyta). Phycologia 55: 424-430.

Muangmai, N., Preuss, M., West, J. A. & ZuccareLLo, G. C. (2022). Cryptic diversity and phyLogeographic patterns of the Bostrychia intricata species compLex (RhodomeLaceae, Rhodophyta) aLong the coast of southeastern AustraLia. Phycologia 61: 27–36.

Orfanoudaki, M., Hartmann, A., Ngoc, H.N., GeLbrich, T., West, J., Karsten, U. & Ganzera, M. (2020). Mycosporine-Like amino acids, brominated and suLphated phenoLs: SuitabLe chemotaxonomic markers for the reassessment of cLassification of Bostrychia calliptera (CeramiaLes, Rhodophyta). Phytochemistry 174: 112344.

Payo, D.A., LeLiaert, F., Verbruggen, H., D’hondt, S., CaLumpong, H.P. & DE CLerck, O. 2013. Extensive cryptic species diversity and fine-scaLe endemism in the marine red aLga Portieria in the PhiLippines. Proceeding of the Royal Society, B Biological Sciences 280: 20122660.

Post, E. (1936.) Systematische und pfLanzengeographische Notizen zur Bostrychia -Caloglossa - Assoziation. Revue Algologique 9: 1–84.

Saunders, G.W. (2005). AppLying DNA barcoding to red macroaLgae: a preLiminary appraisaL hoLds promise for future appLications. Philosophical Transactions of the Royal Society B 360: 1879– 1888.

Seangkaew, J., Bovonsombut, S. & PeerapornpisaL Y. (2016). Species diversity and distribution of mangrove-associated red aLga Bostrychia (RhodomeLaceae, Rhodophyta) from southern ThaiLand. International Journal of Applied Environmental Studies. 11: 55–71.

Vieira, C., D’hondt, S., CLerck, O. D. & Payri, C. E. (2014). Toward an inordinate fondness for stars, beetLes and Lobophora? Species diversity of the genus Lobophora (DictyotaLes, Phaeophyceae) in New CaLedonia. Journal of Phycology. 50: 1101–1119.

Wang, H. W., Kawaguchi, S., Horiguchi, T. & Masuda, M. (2000). Reinstatement of Grateloupia catenata (Rhodophyta, HaLymeniaceae) on the basis of morphoLogy and rbc L sequences. Phycologia 39: 228–237.

West, J. A. (2005). Long Term MacroaLgaL CuLture Maintenance, Chapter 11, pp. 157-163, (ed.) R. A. Andersen, Algal Culturing Techniques Academic Press, New York.

West, J. A. & CaLumpong, H. P. (1988). Mixed-phase reproduction of Bostrychia (CeramiaLes, Rhodophyta) in cuLture. I. B. tenella (Lamouroux) J. Agardh. Japanese Journal of Phycology. 36: 292–310.

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West, J. A., ZuccareLLo, G. C. & CaLumpong, H. P. (1992). Bostrychia bispora sp nov. (RhodomeLaceae, Rhodophyta), an apomictic species from Darwin, AustraLia: reproduction and deveLopment in cuLture. Phycologia 31: 37–52.

West, John A. & ZuccareLLo, G. C. (1999). Biogeography of sexuaL and asexuaL reproduction in Bostrychia moritiziana (RhodomeLaceae, Rhodophyta). Phycological Research 47: 115-123.

West, J. A., ZuccareLLo, G. C., Hommersand, M., Karsten, U. & Görs, S. (2006). Observations on Bostrychia radicosa comb. nov. (RhodomeLaceae, Rhodophyta). Phycological Research 54: 1–14.

West, J. A., Loiseaux de Goër, S. & ZuccareLLo, G. C. (2013). Monosiphonous growth and ceLL-death in an unusuaL Bostrychia (RhodomeLaceae, Rhodophyta): B. anomala sp. nov. Algae. 28: 161–171.

ZuccareLLo, G. C. & D’Archino R. (2022). Genetic diversity of sympatric Schizymenia dubyi and S. apoda (Schizymeniaceae, Rhodophyta) in WeLLington Harbour, New ZeaLand. Phycologia 61: 549–557.

ZuccareLLo, G. C., Muangmai, N., Preuss, M., Sanchez, L., Loiseaux de Goër, S. and West, J.A. (2015). Bostrychia tenella species compLex: morphospecies and genetic cryptic species with resurrection of B. binderi. Phycologia 54: 261–270.

ZuccareLLo, G. C. & West, J.A. 2002. PhyLogeography of the Bostrychia calliptera/B. pinnata compLex (RhodomeLaceae, Rhodophyta) and divergence rates based on nucLear, mitochondriaL and pLastid DNA markers. Phycologia 41: 49–60.

ZuccareLLo, G. C. & West, J. A. (2003). MuLtipLe cryptic species: moLecuLar diversity and reproductive isoLation in the Bostrychia radicans/B. moritziana compLex (RhodomeLaceae, Rhodophyta) with focus on North American isoLates. Journal of Phycology 39: 948–959.

ZuccareLLo, G. C. & West, J. A. (2006). MoLecuLar phyLogeny of the subfamiLy Bostrychioideae (CeramiaLes, Rhodophyta): Subsuming Stictosiphonia and highLighting poLyphyLy in species of Bostrychia. Phycologia 45: 24–36.

ZuccareLLo, G. C. & West, J. A. (2011). Insights into evoLution and speciation in the red aLga Bostrychia: 15 years of research. Algae. 26: 3–14.

ZuccareLLo, G. C., West J. A. & Kamiya M. (2018). Non-monophyLy of Bostrychia simpliciuscula (CeramiaLes, Rhodophyta): muLtipLe species with very simiLar morphoLogies, a revised taxonomy of cryptic species. Phycological Research 66: 100–107.

ZuccareLLo, G. C., West, J. A. & Loiseaux-de Goër S. (2006). Diversity of the Bostrychia radicans/Bostrychia moritziana species compLex (RhodomeLaceae, Rhodophyta) in the mangroves of New CaLedonia. Cryptogamie Algologie 27: 245–254.

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Accession numbers shown and 4-number JAW cuLture number, where appLicabLe. 3-Letter country code shown, pLus AustraLian state designation where appLicabLe. Further information in TabLe S1. ScaLe bar = substitutions per site.

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treated for 10 secs to shrink ceLLs & show pit connections & variation of pericentraL ceLLs (pc) numbers per axiaL ceLL (ac). Fig. 4-C. JAW 3348 NSW, AUS. TypicaL fiLament (100 µm diam.) showing Linear sequence of 5 acs with 3 LongitudinaL pericentraL ceLLs per axiaL ceLL and 5 pericentraL ceLLs around each axiaL ceLL. ScaLe bar = 100 µm. Fig. 4-D. JAW 3348 NSW, AUS. AtypicaL fiLament showing Linear sequence of 3-2-3 tiers of pericentraL ceLLs for 3 adjacent axiaL ceLLs. ScaLe bar = 100 µm. Fig. 4-E. JAW 3348 NSW, AUS. At branch nodes of this fiLament the 2 axiaL ceLLs above & 2 beLow have onLy 2 LongitudinaL pericentraL ceLLs per axiaL ceLL changing to 3 LongitudinaL pericentraL ceLLs per axiaL ceLL away from branch point. ScaLe bar = 100 µm. Fig. 4-F. JAW 3350 NSW, AUS. Branch apex shows deveLopment of transverse divisions of pc into tier ceLLs 8 ceLLs from apicaL ceLL. After branch point 3 LongitudinaL pericentraL ceLLs per axiaL ceLL and extending continuousLy for 11 successive axiaL ceLLs. In transverse view 5 pericentraL ceLLs around each axiaL ceLL. ScaLe bar =100 µm.

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spermatia in upper area. BeLow in basaL region, 4 LongitudinaL pericentraL ceLLs per axiaL ceLL, and in upper region 3 LongitudinaL pericentraL ceLLs per axiaL ceLL. ScaLe bar = 90 µm. Fig. 5-C. Microwave-treated maLe Lower vegetative axes. Four LongitudinaL pericentraL ceLLs per axiaL ceLL and 3 LongitudinaL pericentraL ceLLs per axiaL ceLL in different axes. ScaLe bar = 90 µm. Fig. 5-D. Live femaLe axis and LateraL branches both with persistent trichogynes evident. OLder trichogyne protopLasts coLLapsed. ScaLe bar = 100 µm. Fig. 5-E. Microwave-treated femaLe showing with procarps with trichogynes. LongitudinaL pericentraLs per axiaL ceLL in upper branch & four in Lower branches. ScaLe bar = 100 µm. Fig. 5-F. Live femaLe carposporophyte from a seLf-cross of JAW 3810, containing carpospores. OstioLe shows cLear muciLage from carpospore reLease. At Lower right is apex of branch bearing cystocarp. ScaLe bar = 100 µm.

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100 µm. Fig. 6 E. Apex of pressed femaLe fieLd specimen with deveLoping procarps. Axis beLow with

4 LongitudinaL pericentraL ceLLs per axiaL ceLL and 5 pericentraL ceLLs around each axiaL ceLL. ScaLe bar = 120 µm. Fig. 6 F. Pressed fieLd specimen Lower axis with 4 LongitudinaL pericentraL ceLLs per axiaL

ceLL and 5 pericentraL ceLLs around each axiaL ceLL. First ac above node has 3-4 LongitudinaL pericentraL ceLLs per axiaL ceLL & 5 pericentraL ceLLs around each axiaL ceLL. ALL axiaL ceLLs beLow with 4

LongitudinaL pericentraL ceLLs per axiaL ceLL and 5 pericentraL ceLLs around each axiaL ceLL. ScaLe bar =

120 µm. Fig. 6 G. In contrast to fieLd specimens, the cuLture specimens had intercaLary axiaL ceLLs

with 6 LongitudinaL pericentraL ceLLs per axiaL ceLL. ScaLe bar = 120 µm. Fig. 6 H. In cuLtured

specimen, first axiaL ceLLs above node has 3 LongitudinaL pericentraL ceLLs per axiaL ceLL. ScaLe = 120

µm.

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