Abstract
The carried-out studies concerned two serpentine fern species: Asplenium adulterinum and A. cuneifolium, as well as ferns not connected but also occurring on serpentine rocks, such as: A. septentrionale and Polypodium vulgare. The growth and development of gametophytes of all fern species were affected by Murashige and Skoog (MS) medium composition [Murashige and Skoog (Physiol Plant 15:473–497, 1962)]. Diluted MS medium to one half, and modified MS medium provided the best results in terms of fresh weight increase. The gametophytes growing in these two media (½ MS and MS own modification) had the correct shape and formed the most numerous gametophytes in the first and following rows. Measurements of the fresh weight of gametophytes of A. adulterinum and P. vulgare revealed that growth enhanced with sucrose concentrations between 20 and 30 g L−1. Lower sucrose concentration (5–10 g L−1), decreased the fresh weight of gametophytes and diminished their size. Higher sucrose concentration (40 g L−1) reduced gametophyte growth, though they did not show any discoloration or decay. A. septentrionale tolerated all the sucrose concentrations; it was only without sucrose in the medium that the fresh weight of gametophytes was smaller, as was also their number. The influence of several sugars, such as glucose, fructose, mannose, sucrose, maltose, and lactose (in the same concentration of 0.087 M) on growth and development of gametophytes was studied. The results of investigations showed that each of the fern species studied attained the best increase in the fresh weight, and produced the largest gametophytes in a medium supplemented with sucrose. Application of glucose provided similar results; with the exception of A. adulterinum, which preferred maltose in the medium, and A. cuneifolium, which grew equally well on medium with glucose and mannose. The remaining sugars, and particularly fructose, inhibited the increase of protallial fresh weight or caused its decay. Moreover, fructose turned out to be the most toxic: gametophytes of all studied fern species decayed when fructose was used in the medium.
The studies show that in vitro cultures are an effective method of creating prothallia “gene bank” for serpentine fern species. The best medium for long-term storage of prothallia of A. adulterinum and A. cuneifolium was ½ MS without phytohormones, at low temperature of +8°C, under cool white fluorescent lamp of 0.15 μmol·m−2·s−1.
The number of sporophytes obtained in gametophyte cultures of all the studied species was low. From thousands of gametophytes used, only three to five sporophytes developed. The propagation of sporophytes was tested on media containing kinetin (KIN) and 1-naphtalene acetic acid (NAA). Low doses of these hormones stimulated development of fronds, and higher doses of KIN decreased their size but increased the number of initiated buds. The process of regeneration of new sporophytes was also possible. Rooting of ferns was performed in perlite with a liquid medium, and the influence of diverse soils on the fern survival was evaluated. Plants acclimated well outside in a garden cold frame. The sporophytes reached maturity and produced spores in their second or third year of cultivation.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bogunia, H. and Przywara, L. 1999. Rola cukrowców w roślinnych kulturach in vitro. Wiad. Bot. 43:25–36.
Fernández, H., Bertrand, A., and Sánchez–Tamés, R. 1993. In vitro regeneration of Asplenium nidus L. from gametophytic and sporophytic tissue. Sci. Hortic. 56:71–77
Fernández, H., Bertrand, A., and Sánchez–Tamés, R. 1996a. Micropropagation and phase change in Blechnum spicant and Pteris ansiformis. Plant Cell, Tissue Organ Cult. 44:261–265
Fernández, H., Bertrand, A., and Sánchez–Tamés, R. 1996b. Influence of tissue culture conditions on apogamy in Dryopteris affinis ssp. affinis. Plant Cell, Tissue Organ Cult. 45:93–97
Fernández, H., Bertrand, A., Feito, I., and Sánchez–Tamés, R. 1997a. Gametophyte culture in vitro and antheridiogen activity in Blechnum spicant. Plant Cell, Tissue Organ Cult. 50:71–74
Fernández, H., Bertrand, A., and Sánchez–Tamés, R. 1997b. Gemmation in Osmunda regalis L. gametophyte cultured in vitro. Plant Cell Rep. 16:358–362
Fernández, H., Bertrand, A., and Sánchez–Tamés, R. 1999. Biological and nutritional aspects involved in fern multiplication. Plant Cell, Tissue Organ Cult. 56:211–214
Hirsch, A.M. 1975. The effect of sucrose on the differentiation of excised fern leaf tissue into either gametophytes or sporophytes. Plant Physiol. 56:390–393
Holderegger, R. 1994. Zur Farnflora des Pfannenstils, Kt. Zurich. Farnbl. 25:3–21
Karpowicz, W. 1969. Paprocie. pp. 132–150. Warszawa, PWN
Kempinska, K., Kromer, K., and Zenkteler, E. 2006. Influence of tocopherol on gametogenesis of Asplenium cuneifolium and Asplenium adulterinum. Bot. Guidebooks 29:157–161
Klinkenberg, B. 2008. E-Flora BC: Electronic Atlas of the Plants of British Columbia [www.eflora.bc.ca]. – Lab for Advanced Spatial Analysis, Department of Geography, University of British Columbia, Vancouver
Kromer, K., Marszał-Jagacka, J., Kempińska, K., Nowak, T., Żołnierz, L., Poturała, D., and Świerkosz, K. 2006. In vitro propagation and ex situ preservation of endangered ferns from Lower Silesia. Bot. Guidebooks 29:143–155
Kromer, K., Raj, A., Żołnierz, L., and Poturała, D. 2008. Propagation in vitro and ex situ cultivation of Woodsia alpine (Bolton) S.F. Gray. In: Club Mosses, Horsetails and Ferns in Poland – Resources and Protection, Ed. Szczęśniak E., Gola E., Polish Botanical Society Institute of Plant Biology, University of Wrocław, 2008:15–28
Lovis, J. D. and Reichstein, T. 1968. Űber das spontane Entstehung von Asplenium adulterinum Milde aus einem natűrlichen Bastard. Naturwissenschaften 55(3):117–120
Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bio-assays with tobacco cultures. Physiol. Plant. 15:473–497
Proctor, J. and Woodell, S. R. J. 1971. The plant ecology of serpentine. I. Serpentine vegetation of England and Scotland. J. Ecol. 59(2):375–395
Proctor, J. and Woodell, S. R. J. 1975. The ecology of serpentine soils. In A. Macfadyen ed. Advances in Ecological Research, pp. 255–366. Academic, London.
Raghavan, V. 1989. Developmental biology of fern gametophytes. Cambridge University Press, New York.
Reichstein, T. 1984. Aspleniaceae. In: HEGI G., Illustriertè Flora von mitteleuropa. 3 Aufl. 1.1:211–275
Reiger, R., Michaelis, A., and Green, M. M. 1976. Glossary of Genetic and Cytogenetics. Springer, Berlin
Roberts, B. A. and Proctor, J. 1992. The ecology of areas with serpentinized rocks. A word view. Kluwer Academic, Dordrecht, Boston, London, 427 pp.
Romano, A., Noronha, C., and Martinis-Loncao, M.A. 1995. Role of carbohydrate in micropropagation of cork oak. Plant Cell, Tiss. Organ Cult. 40:159–167
Sarosiek, J. and Sadowska, A. 1961. Ekologia roślin gleb serpentynowych. Wiad. Bot. 5:73–86
Somer, M., Arbesú R., Menéndez, V., Revilla, M. A., and Fernández H. 2010. Sporophyte induction studies in ferns in vitro. Euphytica 171:203–210
Szczęśniak, E., 2006. Asplenium serpentine ferns in Poland – threats and conservation imperatives. Bot. Guidebooks 29:89–98
Whittaker, R. H. 1954. The vegetational response to serpentine soils. Ecology 35:275–288
Zarzycki, K., and Kaźmierczakowa, R. 2001. Polska Czerwona Księga Roślin. Paprotniki i Rośliny Kwiatowe. PAN, Inst. Bot. Im. W. Szafera i Inst. Ochrony Przyrody, Kraków
Zenkteler, E. 1992. Metoda in vitro w rozmnażaniu i okresowym przechowywaniu chronionych oraz rzadkich i ginących gatunków paproci. Hod. Rośl. Nasien. 5:20–30
Zenkteler, E. 2000. Systemy wegetatywnego rozmnażania paproci in vivo oraz in vitro. Wydawnictwo Naukowe UAM, Poznań
Żołnierz, L., Kromer, K., and Świerkosz, K. 2008. Ladder spleenwort (Asplenium adulterinum Milde) in Poland – distribution, population state and conservation plan framework. In: Club Mosses, Horsetails and Ferns in Poland – Resources and Protection, Ed. Szczęśniak E., Gola E., pp. 29–45. Polish Botanical Society Institute of Plant Biology, Wrocław, University of Wrocław
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this chapter
Cite this chapter
Marszał-Jagacka, J., Kromer, K. (2011). In Vitro Propagation of Rare and Endangered Serpentine Fern Species. In: Kumar, A., Fernández, H., Revilla, M. (eds) Working with Ferns. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-7162-3_11
Download citation
DOI: https://doi.org/10.1007/978-1-4419-7162-3_11
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4419-7161-6
Online ISBN: 978-1-4419-7162-3
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)