Abstract
Medusagyne oppositifolia is a critically endangered monotypic species with only a few trees left in the wild. Due to invasion and habitat clearance, genetic diversity of this species has been reduced to an alarmingly low level. Long-term in vitro cultures of M. oppositifolia were slow-growing and eventually lost the ability to multiply and root. Because of the lack of new seed and vegetative materials, efforts were made to rejuvenate the existing cultures. This was attempted by a series of steps involving serial pruning of the shoot tips and growing on a modified MS medium containing silver nitrate and thidiazuron. Rapid cycling clonal propagules were produced this way from cultured nodes. Isolated shoots were rooted on Florialite™, and rooted plants were weaned in a transition stage using a Nutrient Film Technique (NFT) hydroponic system before being completely weaned under glasshouse conditions. The importance of screening cultures for multiplication and rooting efficiency during long-term maintenance is discussed in the context of stress metabolite production and its influence on growth in culture. Factors contributing to rejuvenation of old cultures, application of unconventional supporting systems for rooting and application of the hydroponic system as a weaning tool are also discussed.
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References
Banerjee S.; Tripathi J.; Verma P. C.; Dwivedi P. D.; Khanuja S. P. S.; Bagchi G. D. Thidiazuron-induced high-frequency shoot proliferation in Cineraria maritima Linn. Curr Sci 87: 1287–1290; 2004.
Dickison W. C. The morphology and relationships of Medusagyne (Medusagynaceae). Plant Syst Evol 171: 27–55; 1990.
Dixon K. W. Towards integrated conservation of Australian endangered plants - the Western-Australian model. Biodivers Conserv 3: 148–159; 1994.
Fay M. F.; Swensen S. M.; Chase M. W. Taxonomic affinities of Medusagyne oppositifolia (Medusagynaceae). Kew Bulletin 52: 111–120; 1997.
Gaspar T.; Kevers C.; Bisbis B.; Franck T.; Crevecoeur M.; Greppin H.; Dommes J. Loss of plant organogenic totipotency in the course of in vitro neoplastic progression. In Vitro Cell Dev-Pl. 36: 171–181; 2000.
Gibson A. C.; Nobel P. S. The cactus prime. Harvard University Press, Cambridge; 1986.
Giridhar P.; Reddy B. O.; Ravishankar G. A. Silver nitrate influences in vitro shoot multiplication and root formation in Vanilla planifolia Andr. Curr Sci 81: 1166–1170; 2001.
Graham E. T.; Joshi P. A. Plant cuticle staining with bismarck brown Y and azure B or toluidine blue O before paraffin extraction. Biotech Histochem 71: 92–95; 1996.
Hanson L.; Boyd A.; Johnson M. A. T.; Bennett M. D. First nuclear DNA C-values for 18 eudicot families. Ann Bot 96: 1315–1320; 2005.
Hoagland D. R.; Arnon D. I. The water-culture method for growing plants without soil. Circ Calif Agric Exp Station 347: 32; 1950.
Ibrahim R.; Debergh P. C. Improvement of adventitious bud formation and plantlet regeneration from in vitro leaflet explants of roses (Rosa hybrida L.). Acta Hortic 520: 271–279; 2000.
IUCN: IUCN Red List of Threatened Plants. The World Conservation Union, Gland, Switzerland and Cambridge, UK; 1998
Kauth P.; Kane M. E.; Vendrame W. Greenhouse irrigation method influences growth and quality of in vitro propagated Cryptocoryne wendtii plantlets. Plant Cell, Tissue Organ Cult 87: 219–222; 2006.
Kumar P. P.; Lakshmanan P.; Thorpe T. A. Regulation of morphogenesis in plant tissue culture by ethylene. In Vitro Cell Dev-Pl 34: 94–103; 1993.
Mccown B. H. Micropropagation of hardy rose species and hybrids. Hortscience 15: 417–417; 1980.
McCown B. H. Recalcitrance of woody and herbaceous perennial plants: dealing with genetic predeterminism. In Vitro Cell Dev-Pl 36: 149–154; 2000.
Murashige T.; Skoog F. A. Revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plantarum 15: 473–497; 1962.
O’Brian T.; McCully M. The study of plant structure:principles and selected methods. Termacarphi, Melbourne; 1981.
Pence V. C. Cryopreservation of in vitro grown fern gametophytes. Am Fern J 90: 16–23; 2000.
Robertson S. A. Flowering plants of Seychelles. University of Chicago Press, Chicago; 1989.
Rosu A.; Skirvin R. M.; Bein A.; Norton M. A.; Kushad M.; Otterbacher A. G. The development of putative adventitious shoots from a chimeral thornless Rose (Rosa multiflora Thunb Ex J Murr) in vitro. J Hortic Sci 70: 901–907; 1995.
Sarasan V. Application of Sorbarods and Florialite to rooting of critically endangered tree species Trochetiopsis ebenus. Acta Hortic 616: 211–214; 2003.
Sharma N.; Chandel K. P. S. Low-temperature storage of Rauwolfia serpentina Benth Ex Kurz - an endangered, endemic medicinal Plant. Plant Cell Rep 11: 200–203; 1992.
Van der Salm T. P. M.; Van der Toorn C. J. G.; Tencate C. H. H.; Dubois L. A. M.; De Vries D. P.; Dons H. J. M. Importance of the iron chelate formula for micropropagation of Rosa hybrida L ‘Moneyway’. Plant Cell, Tissue Organ Cult 37: 73–77; 1994.
YongHua Q.; ShangLong Z.; Xiao Z. L.; Yu Z. D.; Syed A. Response of in vitro strawberry to silver nitrate (AgNO3). Hortscience 40: 747–751; 2005.
Acknowledgements
The authors gratefully acknowledge Prof. Andy Roberts for the critical reading and useful suggestions, Lynda Hanson of Jodrell laboratories for assistance in flow cytometry, Dr. Peter Gasson for advice on histochemistry and Chris Haysom and Nick Johnson for growing weaned plants in RBG Kew glasshouses.
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Editor: P. Lakshmanan
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Marriott, P., Sarasan, V. Novel micropropagation and weaning methods for the integrated conservation of a critically endangered tree species, Medusagyne oppositifolia . In Vitro Cell.Dev.Biol.-Plant 46, 516–523 (2010). https://doi.org/10.1007/s11627-010-9321-8
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DOI: https://doi.org/10.1007/s11627-010-9321-8