In vitro cultivation and propagation strategies in Bryaceae (Bryophyta): A review
DOI:
https://doi.org/10.14719/tcb.9458Keywords:
Bryaceae, growth media, in vitro culture, moss propagation, phytohormonesAbstract
Bryophytes, particularly mosses of the family Bryaceae, represent an evolutionarily significant group with considerable medicinal potential. Despite their ecological, evolutionary and pharmacological importance, in vitro studies on bryophytes remain limited when compared to those on vascular plants. To address this gap, the present review synthesizes existing protocols and methodologies for the in vitro propagation of Bryaceae, with a focus on species prevalent in India. It highlights key factors influencing successful culture, including sterilization techniques, temperature, photoperiod, culture vessels, media composition, sugar types and hormonal supplements. Sodium hypochlorite has emerged as the most effective sterilizing agent, while optimal growth temperature ranges from 22–27 °C. Standard photoperiods of 16/8 and 12/12 hr light/dark have been widely adopted. Petri plates are the preferred culture vessels due to ease of handling and observations. Among growth media, Knop and Murashige and Skoog media, sometimes in modified formulations, have shown the highest efficacy for protonema and gametophyte development across species like Bryum argenteum, Rhodobryum giganteum and Anomobryum filiforme. Hormonal responses are highly concentration-dependent and exhibit interspecific variability. Along with plant growth regulators, auxin and cytokinin are the two principal hormones that play a critical role in regulating bud initiation and gametophyte development. Additionally, sugars such as glucose and sucrose promote growth in certain Bryaceae species but may inhibit others. This review highlights critical research gaps in the understanding of light intensity and spectral quality (i.e., specific wavelength suitable for growth and development), temperature tolerance and nutrient formulations, underscoring the need for species-specific in vitro protocols. The findings will lay a foundation for advancing Bryaceae propagation for conservation and bio-industrial applications.
Downloads
References
Dandotiya DI, Govindapyari H, Suman S, Uniyal PL. Checklist of the bryophytes of India. Archive for bryology. 2011;88(1):1-26.
Spence J.R. Bryum Hedw.(Bryaceae). Vol. 28. Flora North America; 2014. p. 124–129 p.
Choudhary BL, Sanadhya C, Sharma TP. Bryophyte Flora of Gujarat India. Udaipur: Himanshu Publications; 2006. p. 197 .
Asakawa Y. Chemical constituents of the bryophytes. In: Progress in the chemistry of organic natural products. Vienna: Springer Vienna; 1995. p. 1–562. https://cir.nii.ac.jp/crid/1573950399095172480.bib?lang=en
Asakawa Y. Biologically active substances from bryophytes. In: Bryophyte Development. CRC Press; 2019 Jun 4. p. 259–288.
Mishra R, Pandey VK, Chandra R. Potential of bryophytes as therapeutics. International Journal of Pharmaceutical Sciences and Research. 2014;5(9):3584. http:// doi.org/10.13040/IJPSR.0975-8232.5
Glime J. Bryophyte Ecology: Volume 5: Uses, 2019.
Servettaz C.Recherches expérimentales sur le dévelopment et la nutrition des mousses en milieux stérilise. Ann Sci Nat Bot Bio. 1913;17:111–223.
Reski R. Development, genetics and molecular biology of mosses. Botanica Acta. 1998;111:1–15.
Duckett J.G., Schmid A.M., Ligrone R. Protonemal morphogenesis. In: Bates J.W., Ashton N.W., Duckett J.G., editors. Bryology for the twenty-first century. First. London: Maney Publishing and the British Bryological Society; 1998. p. 223–46.
Newton AE, Cox CJ, Duckett JG, Wheeler JA, Goffinet B, Hedderson TA, Mishler BD. Evolution of the major moss lineages: phylogenetic analyses based on multiple gene sequences and morphology. The bryologist. 2000 Jun;103(2):187-211. https://doi.org/10.1639/0007-2745(2000)103[0187:EOTMML]2.0.CO;2
Buckley P.M., Reed B.M. Antibiotic susceptibility of plant-associated bacteria. HortScience. 1994;434c–434. https://doi.org/10.21273/HORTSCI.29.5.434c
Vashishta B. R., Sinha A. K., Adarsh Kumar. Botany For Degree Students - Bryophyta. S Chand; 2010.
Sahu V, Asthana AK. An observation on growth response of Anomobryum filiforme var. concinnatum (Spruce) Aman. (Bryaceae) in different culture media. National Academy Science Letters. 2013 Dec;36(6):587–9. https://doi.org/10.1007/s40009-013-0173-8
da Costa Silva-e-Costa J, Luizi-Ponzo AP, de Resende CF, Peixoto PHP. Spore germination, early development and some notes on the effects of in vitro culture medium on Frullania ericoides (Nees) mont. (Frullaniaceae, marchantiophyta). Acta Bot Brasilica. 2017;31(1):19–28. https://doi.org/10.1590/0102-33062016abb0336
Liang SF, Sun Y, Zhu RL. In vitro micropropagation of Bryum argenteum Hedw. Cryptogamie Bryologie. 2010;31(3):233–9.
Post AR, McCall DS, Askew SD. Preemergence control of silvery threadmoss (Bryum argenteum) grown from spores and bulbils in axenic culture. Weed Technology. 2016;30(1):198-206. https://doi.org/10.1614/WT-D-14-00125.1
Bijelovi? A, Sabovljevi? M, Grubisi? D, Konjevi? R. Phytohormone influence on the morphogenesis of two mosses (Bryum argenteum Hedw. and Atrichum undulation (Hedw.) P. Beauv.). Isr J Plant Sci. 2004;52(1):31–6. https://doi.org/10.1560/RX30-U7QH-DEMQ-GL7P
Sabovljevi? MS, Bijelovi? A, Dragi?evi? I?. Effective and easy way of establishing in vitro culture of mosses, Bryum argenteum Hedw. and Bryum capillare Hedw.-Bryaceae. Archives of Biological Sciences. 2002;54(1-2):7-8. https://doi.org/10.2298/ABS0202001S
Sabovljevic A, Sabovljevic M, Grubisic D, Konjevic R. The effect of sugars on development of two moss species (Bryum argenteum and Atrichum undulatum) during in vitro culture. Belgian journal of botany. 2005 :79-84.
Chen YY, Lou YX, Guo SL, Cao T, Chen YY, Lou YX, et al. Successful tissue culture of the medicinal moss Rhodobryum giganteum and factors influencing proliferation of its protonemata. Ann Bot Fennici. 2009;46:516–24. https://doi.org/10.5735/085.046.0604
Proctor MCF. The Diversification of bryophytes and vascular plants in evolving terrestrial Environments; 2014. p. 59–77.
Kallio P. Heinonen Saini. Ecology of Rhacomitrium lanuginosum (Hedw.) Brid. Rep. Kevo Subarctic Res. 1973;10:43-54.
Weis E, Wamper D, Santarius KA. Heat sensitivity and thermal adaptation of photosynthesis in liverwort thalli. Oecologia. 1986;69(1):134-9. https://doi.org/10.1007/BF00399049
Gabriel R.M.De Almeida. Ecophysiology of Azorean Forest Bryophytes. [Ph.D Thesis]. Imperial College of Science, Technology and Medicine; 2000.
Chopra RN, Rawat MS. Studies on production and behavior of protonemal gemmae in some Bryaceae. Bryologist. 1977;80.
Tixier P. Bryogeographie du Mont Bokor (Cambodge). Vol. 18. Bryophytorum Bibliotheca; 1979. p. 1–121.
Sabovljevi? A, Sabovljevi? M, Grubiši? D. Gibberellin influence on the morphogenesis of the moss Bryum argenteum Hedw. In in vitro conditions. Arch Biol Sci. 2010;62(2):373–80. https://doi.org/10.2298/ABS1002373S
Silva AS, Pôrto KC, Simabukuro EA. Effects of light and nutrients on different germination phases of the cosmopolitan moss Bryum argenteum Hedw. (Bryaceae). Brazilian Archives of biology and technology. 2010;53:763-9. https://doi.org/10.1590/S1516-89132010000400003
Cove DJ, Perroud PF, Charron AJ, McDaniel SF, Khandelwal A, Quatrano RS. Culturing the moss Physcomitrella patens. Cold Spring Harb Protoc. 2009;4(2). https://doi.org/10.1101/pdb.prot5144
Beneke W. Uber die Keimung der Brutknospen von Lunularia cruciata. Botanische Zeitung. 1903;61:19–46.
Marchal L.E., Marchal E. Recherches physiologiques sur l’amidon chez les Bryophytes. Bulletin of the Société royale de botanique de Belgique. 1906;43:115–214.
Sabovljevic M, Papp B, Sabovljevic A, Vujicic M, Szurdoki E, Segarra-Moragues JG. In vitro micropropagation of rare and endangered moss Entosthodon hungaricus (Funariaceae). Biosci. J., Uberlândia. 2012;28(4):632-640.
Pressel S, Matcham HW, Duckett JG. Studies of protonemal morphogenesis in mosses. XI. Bryum and allied genera: A plethora of propagules. J Bryol. 2007;29(4):241–58. https://doi.org/10.1179/174328207X244042
Bopp M. Developmental physiology of bryophytes. In: Schuster R.M, editor. New manual of bryology. First. Japan: Hattori Botanical Laboratory; 1993. p. 276–324.
Goode JA, Duckettj JG, Steadj AD. Protonemal Morphogenesis of the Moss Tetraphis pellucida Hedw. in Culture and in the Wild. Vol. 70, Annals of Botany. 1992. https://doi.org/10.1093/oxfordjournals.aob.a088512
Duckett JG, Goode JA, Matcham HW. Studies of protonemal morphogenesis in mosses. VIII. The gemmiferous protonemata of Orthodontium and Dicranoweisia. J Bryol. 2001;23(3):181–93. https://doi.org/10.1179/jbr.2001.23.3.181
Duckett J.G., Burch J., Fletcher P.W., Matcham H.W., Read D.J., Russell A.J., et al. In vitro cultivation of bryophytes: a review of practicalities, problems, progress and promise. J Bryol. 2004;26(1):3–20.
Simola LK. The effect of several protein amino acids and some inorganic nitrogen sources on the growth of Sphagnum nemoreum. Physiol Plant. https://doi.org/10.1111/j.1399-3054.1975.tb03892.x
Kajita M, Takio S, Takami S, Hino S. Establishment and growth characterization of suspension culture of cells from the moss, Sphagnum imbrication. Physiologia plantarum. 1987;70(1):21-6. https://doi.org/10.1111/j.1399-3054.1987.tb08691.x
Beutelmann P., Bauer L. Purification and identification of a cytokinin from moss callus cells. Planta. 1977;133(3):215–217. https://doi.org/10.1007/BF00380679
Christianson M.L. Control of morphogenesis in bryophytes. In: Shaw A.J., Goffinet B., editors. Bryophyte Biology. First. Cambridge: Cambridge University Press; 2000. p. 199–224.
Bhatla SC, Chopra RN. Hormonal regulation of gametangial formation in the moss Bryum argenteum Hedw. Journal of Experimental Botany. 1981;32(6):1243-56. https://doi.org/10.1093/jxb/32.6.1243
Goode JA, Stead AD, Duckett JG. Studies of protonemal morphogenesis in mosses II Orthotrichum obtusifolium Brid. Journal of bryology. 1993;17(3):409-19. https://doi.org/10.1179/jbr.1993.17.3.409
Hassid WZ. Transformation of sugars in plants. Annual Review of Plant Physiology. 1967;18(1):253-80. https://doi.org/10.1146/annurev.pp.18.060167.001345
Gibson SI. Sugar and phytohormone response pathways: navigating a signalling network. Journal of experimental botany. 2004;55(395):253-64. https://doi.org/10.1093/jxb/erh048 43.
Smeekens S. Sugar-induced signal transduction in plants. Annual review of plant biology. 2000;51(1):49-81. https://doi.org/10.1146/annurev.arplant.51.1.49
Downloads
Published
Issue
Section
License
Copyright (c) 2026 Mehta D, Gujar R V
This work is licensed under a Creative Commons Attribution 4.0 International License.
