Skip to main content

Advertisement

SpringerLink
Log in

Encapsulation of moss buds: an efficient method for the in vitro conservation and regeneration of the endangered moss Splachnum ampullaceum

  • Original Paper
  • Published:
Plant Cell, Tissue and Organ Culture Aims and scope Submit manuscript

Abstract

In vitro culture techniques are usually employed for ex situ conservation of endangered plant species. However, encapsulation to preserve threatened bryophytes is scarcely used, and only as a pretreatment prior to cryopreservation. In our study, two different methods of germplasm conservation, involving calcium-alginate encapsulation of moss material, were assessed. The plant material used was gametophyte buds (gametophores) of Splachnum ampullaceum Hedw., a rare species of moss. Moss regeneration was evaluated at different periods of time to examine the efficacy of the technique for moss germplasm conservation. The effects of encapsulation and cold storage on developmental parameters such as protonematal colony diameter, bud length, and number of buds were also studied. The results suggest that moss encapsulation with no prior treatment may be a suitable method for germplasm conservation during long periods of time. With our method survival rates as high as 50% could be reached after 2.5 years of cold storage versus 0% of 24-h cryopreserved beads. This technique together with cold storage, avoiding freezing, may be especially important in desiccation intolerant mosses.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Arrabal R, Amancio F, Carneiro LA, Neves LJ, Mansur E (2002) Micropropagation of endangered endemic Brazilian bromeliad Cryptanthus sinuosus (LB Smith) for in Vitro preservation. Biodiv Conserv 11(6):1081–1089

    Article  Google Scholar 

  • Bajaj YPS (ed) (1995a) Somatic embryogenesis and synthetic seed I. In: Biotechnology in agriculture and forestry, vol 30. Springer, Berlin-Heildelberg-New York

  • Bajaj YPS (ed) (1995b) Somatic embryogenesis and synthetic seed II. In: Biotechnology in agriculture and forestry, vol 31. Springer, Berlin-Heildelberg-New York

  • Benson EE, Danaher JE, Pimbley IM, Anderson CT, Wake JE, Daley S, Adams LK (2000) In vitro micropropagation of Primula scotica: a rare Scottish plant. Biodiv Conserv 9(7):711–726

    Article  Google Scholar 

  • Blandino A, Macías M, Cantero D (2000) Glucose oxidase release from calcium alginate gel capsules. Enz Mic Tech 27:319–324

    Article  CAS  Google Scholar 

  • Block W (2003) Water status and thermal analysis of alginate beads used in cryopreservation of plant germplasm. Cryobiol 47:59–72

    Article  CAS  Google Scholar 

  • Bunn E (2005) Development of in vitro methods for ex situ conservation of Eucalyptus impensa, an endangered mallee from southwest Western Australia. Plant Cell Tiss Org Cult 83(1):97–102

    Article  CAS  Google Scholar 

  • Burch J (2003) Some mosses survive cryopreservation without prior pretreatment. Bryologist 106(2):270–277

    Article  Google Scholar 

  • Burch J, Wilkinson T (2002) Cryopreservation of protonemata of Ditrichum cornubicum (Paton) comparing the effects of four cryoprotectant pretreatments. Cryo Letters 23:197–208

    PubMed  CAS  Google Scholar 

  • Cameron R, Wyatt R (1986) Substrate restriction in entomophilous Splachnaceae. Role of spore dispersal. Bryologist 89(4):279–284

    Google Scholar 

  • Carneiro LA, Araújo RFG, Brito GMJ, Fonseca MHPB, Costa A, Crocomo OJ, Masur E (1998) In vitro regeneration from leaf explants of Neoregelia cruenta (R. Graham) L.B. Smith, an endemic bromeliad from Eastern Brazil. Plant Cell Tiss Org Cult 55(2):79–83

    Article  Google Scholar 

  • Christianson M (1998) A simple protocol for cryopreservation of mosses. Bryologist 101:32–35

    Google Scholar 

  • Da Costa Nunes E, Benson EE, Oltramari AC, Araujo PS, Moser JR, Viana AM (2003) In Vitro conservation of Cedrela fissilis Vellozo (Meliaceae), a native tree of the Brazilian Atlantic forest. Biodiv Conserv 12(4):837–848

    Article  Google Scholar 

  • Draper D, Rosselló-Graell A, Garcia C, Tauleigne Gomes C, Sérgio C (2003) Application of GIS in plant conservation programmes in Portugal. Biol Conserv 113:337–349

    Article  Google Scholar 

  • Duckett JG, Burch J, Fletcher PW, Matcham HW, Read DJ, Russel AJ, Pressel S (2004) In vitro cultivation of bryophytes: a review of practicalities, problems, progress and promise. J Bryol 26(1):3–20

    Article  Google Scholar 

  • ECCB (1995) Red Data Book of European Bryophytes. European Committee for Conservation of Bryophytes (ECCB), Trondheim

    Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  PubMed  CAS  Google Scholar 

  • George EF (1993) Plant propagation by tissue culture. Exegetics Limited, Edington

    Google Scholar 

  • Gonçalves S, Romano A (2005) Micropropagation of Drosophyllum lusitanicum (Dewy pine), an endangered West Mediterranean endemic insectivorous plant. Biodiv Conserv 14(5):1071–1081

    Article  Google Scholar 

  • González ML, Mallón R, Reinoso J, Rodríguez-Oubiña J (2006) In Vitro micropropagation and long-term conservation of the endangered moss Splachnum ampullaceum Hedw. Biol Plant 50(3):339–345

    Article  Google Scholar 

  • González-Benito ME, Pérez C, Viviani AB (1997) Cryopreservation of nodal explants of an endangered plant species (Centaurium rigualii Esteve) using the encapsulation-dehydration method. Biodiv Conserv 6(4):583–590

    Article  Google Scholar 

  • Hallingbäck T, Hodgetts N (2000) Status Survey and Conservation Action Plan for Bryophytes: Mosses, Liverworts and Hornworts. In: IUCN / SSC Bryophyte Specialist Group. IUCN, Gland-Switzerland-Cambridge, p 106

  • Heller R (1953) Researches on the mineral nutrition of plant tissues. Ann Sci Nat Bot Biol Vég 11thSer:141–223

    Google Scholar 

  • Hugonnot V, Bardat J (2001) Données sur l’ecológie de quatre bryophytes rares dans le Massif Central. J Bot Soc France 14:37–40

    Google Scholar 

  • Ipekei Z, Gozukirmizi N (2003) Direct somatic embryogenesis and synthetic seed production from Paulownia elongata. Plant Cell Rep 22:16–24

    Article  Google Scholar 

  • Kulseng B, Skjak-Braek G, Ryan L, Andersson A, King A, Faxvaag A, Espevik T (1999) Transplantation of alginate microcapsules: generation of antibodies against alginates and encapsulated porcine islet-like cell clusters. Transplantation 67:978–984

    Article  PubMed  CAS  Google Scholar 

  • Lisek A, Orlikowska T (2004) In vitro storage of strawberry and raspberry in calcium-alginate beads at 4°C. Plant Cell Tiss Org Cult 78(2):167–172

    Article  CAS  Google Scholar 

  • Mallón R, Reinoso J, Rodríguez-Oubiña J, González ML (2006) In vitro development of vegetative propagules in Splachnum ampullaceum: brood cells and chloronematal bulbils. Bryologist 109(2):215–223

    Article  Google Scholar 

  • Martin KP, Pradeep AK (2003) Simple strategy for the in vitro conservation of Ipsea malabarica an endemic and endangered orchid of the Western Ghats of Kerala, India. Plant Cell Tiss Org Cult 74(2):197–200

    Article  Google Scholar 

  • Nadeem M, Palni LMS, Purohit AN, Pandey H, Nandi SK (2000) Propagation and conservation of Podophylum hexandrum Royle: an important medicinal herb. Biol Conserv 92:121–129

    Article  Google Scholar 

  • Nagao M, Minami A, Arakawa K, Fujikawa S, Takezawa D (2005) Rapid degradation of starch in chloroplasts and concomitant accumulation of soluble sugars associated with ABA-induced freezing tolerance in the moss Physcomitrella patens. J Plant Physiol 162:169–180

    Article  PubMed  CAS  Google Scholar 

  • Patel AV, Pusch I, Mix-Wagner G, Vorlop KD (2000) A novel encapsulation technique for the production of artificial seeds. Plant Cell Rep 19:868–874

    Article  CAS  Google Scholar 

  • Pence VC (1998) Cryopreservation of Bryophytes: the effects of abscisic acid and encapsulation dehydration. Bryologist 10(2):278–281

    Google Scholar 

  • Pourjavadi A, Barzegar Sh, Mahdavinia GR (2006) MBA-crosslinked Na-Alg/CMC as a smart full-polysaccharide superabsorbent hydrogels. Carbohyd Polym 66(3):386–395

    Article  CAS  Google Scholar 

  • Proctor MFC (2003) Experiments on the effect of different intensities of desiccation on bryophyte survival, using chlorophyll fluorescence as an index of recovery. J Bryol 25:201–210

    Google Scholar 

  • Rech Filho A, Dal Vesco L, Nodari RO, Lischka RW, Mueller CV, Guerra MP (2005) Tissue culture for the conservation and mass propagation of Vriesea reitzii Leme and Costa, a bromeliad threatened of extinction from the Brazilian Atlantic Forest. Biodiv Conserv 14(8):1799–1808

    Article  Google Scholar 

  • Redenbaugh K, Fujii J, Slade D, Viss P, Kossler M (1991) Artificial seeds-Encapsulated somatic embryos. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, high-tech and micropropagation. Springer-Verlag, Heidelberg, pp 395–416

    Google Scholar 

  • Reinoso J, Rodríguez-Oubiña J, Viera MC (2002) Lista Roja de los Briófitos de Galicia (NO de España). Nova Acta Cient. Compostelana (Biol.) 12:83–93

    Google Scholar 

  • Sabovljevic M, Bijelovic A, Dragicevic I (2003) In vitro Culture of Mosses: Aloina aloides (K.F.Schultz) Kindb., Brachythecium velutinum (Hedw.) B.S., G. and Grimmia pulvinata (Hedw.) Sm. Turk J Bot 27:441–446

    Google Scholar 

  • Schofield WB (1985) Introduction to bryology. Macmillan, New York

    Google Scholar 

  • Schulte J, Reski R (2004) High throughput cryopreservation of 140000 Physcomitrella patens mutants. Plant Biol 6:119–127

    Article  PubMed  CAS  Google Scholar 

  • Sérgio C, Casas C, Brugués M, Cros RM (1994) Red list of bryophytes of the Iberian Peninsula. ICN, Lisboa, pp 1–50

    Google Scholar 

  • Shaw AJ, Goffinet B (2000) Bryophyte biology. Cambridge University Press, Cambridge, pp 1–476

    Google Scholar 

  • Söderström L, Hallingbäck T, Gustafsson L, Cronberg N, Hedenäs L (1992) Bryophyte conservation for the future. Biol Conserv 59:265–270

    Article  Google Scholar 

  • SPSS Inc. (2002) SPSS Version 11.5.1 (Computer program). SPSS Inc., Chicago

Download references

Acknowledgements

This work was financially supported by the Xunta de Galicia, Spain (project no. PGIDT00MAM20302PR). The authors sincerely thank Dr. José Blanco for his help on the encapsulation protocols. We thank Dr. Juan Reinoso and Dr. Juan Rodríguez-Oubiña for supplying us with the wild specimen. We also acknowledge Dr. Jaime Gómez-Márquez and Dr. Luisa Areses for their critical revision of the manuscript. We thank Christine Francis for her help with the English translation. We also thank referees for their helpful suggestions.

Author information

Author notes

  1. Rubén Mallón & María Luz González

    Present address: Departamento de Fisiología Vegetal, Facultad de Biología, Universidad de Santiago de Compostela, 15782, Santiago de Compostela, A Coruña, Spain

  2. Asteria Luzardo

    Present address: Facultad de Farmacia, Universidad de Santiago de Compostela, 15782, Santiago de Compostela, Spain

  3. Paula Barros

    Present address: Departamento de Bioquímica y Biología Fundamental, Facultad de Biología, Universidad de Santiago de Compostela, 15782, Santiago de Compostela, Spain

Authors and Affiliations

  1. Departamento de Fisiología Vegetal, Universidad de Santiago de Compostela, Santiago de Compostela, Spain

    Rubén Mallón, Paula Barros & María Luz González

  2. Departamento de Farmacia y Tecnología Farmacéutica, Universidad de Santiago de Compostela, Santiago de Compostela, Spain

    Asteria Luzardo

Authors
  1. Rubén Mallón
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Paula Barros
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Asteria Luzardo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. María Luz González
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to María Luz González.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mallón, R., Barros, P., Luzardo, A. et al. Encapsulation of moss buds: an efficient method for the in vitro conservation and regeneration of the endangered moss Splachnum ampullaceum . Plant Cell Tiss Organ Cult 88, 41–49 (2007). https://doi.org/10.1007/s11240-006-9176-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-006-9176-1

Keywords

Search

Navigation