Abstract
Shoot tips and nodes from a genotype of Cistus clusii were cultured on a medium containing Murashige and Skoog macronutrients, Nitsch and Nitsch micronutrients, sucrose, iron, thiamine, myoinositol, and agar. This establishment medium, enriched with growth regulators and the biocide substances Plant Preservative Mixture and Thiabendazole lactate, improved explant survival by 14–16% and reduced contamination late in culture. For the proliferation stage, the explants rapidly formed axillary buds on a culture medium containing 6-benzylaminopurine (0.5 mg l−1). The best response for rooting was obtained on a culture medium with a 0.1 mg l−1 indolebutyric acid supplement. Rooted plantlets were acclimatized to greenhouse conditions and then transferred to the field in order to evaluate their phenotypic homogeneity. Karyotyping showed that the in vitro propagated plantlets have the same chromosome numbers as the mother plants. The success of this work indicates that micropropagation can be a useful tool for the conservation of C. clusii Dunal, an endangered plant in Italy.
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Almeida R.; Gonçalves S.; Romano A. In vitro micropropagation of endangered Rhododendron ponticum L. subsp. baeticum (Boissier & Reuter) Handel-Mazzetti. Biodiversity Conserv. 14: 1059–1069; 2005. doi:10.1007/s10531-004-8413-3.
Arrabal R.; Amancio F.; Carneiro L. A.; Neves L. J.; Mansur E. Micropropagation of endangered endemic Brazilian bromeliad Cryptanthus sinuosus (L.B. Smith) for in vitro preservation. Biodiversity Conserv. 11: 1081–1089; 2002. doi:10.1023/A:1015860804695.
Benson, E. E.; Danaher, J. E.; Pimbley, I. M.; Anderson, C. T.; Wake, J. E.; Daley, S.; Adams, L. K. In vitro micropropagation of Primula scotica: a rare Scottish plant. Biodiversity Conserv. 9: 711–726; 2000. doi:10.1023/A:1008941726419.
Cavallaro V.; Forte L.; Macchia F.; Pastore D.; Scatigna A. M. E. Ecology of germination. In: Macchia F. (ed) Biology and ecology of Cistus clusii Dunal. Editoriale Adda, Bari, Italy, pp 17–22; 2005. Reference translated from Italian language.
Chiesura Lorenzoni F.; Curti L.; Lorenzoni G. G.; Marchiori S. Cistus clusii Dunal. Nomenclature and distribution notes with particular regard at Lesina (Foggia) botanical station. Giorn. Bot. Ital. 110: 259–267; 1976. Reference translate by Italian language.
Conti F.; Manzi A.; Pedrotti F. The red book of Italian plants. Ministero dell’Ambiente–WWF, Italy; 1992. Reference translated from Italian language.
Conti F.; Manzi A.; Pedrotti F. Regional red lists of plants. WWF–Italy, Italian Botanical Society, Società Botanica Italiana, University of Camerino, Italy; 1997. Reference translated from Italian language.
D'Emerico S. Comparative karyological analysis between Lesina and Spain population. In: Macchia F. (ed) Biology and ecology of Cistus clusii Dunal. Editoriale Adda, Bari, Italy, pp 39–42; 2005. Reference translated from Italian language.
Edson J. L.; Leege-Brusven A. D.; Everett R. L.; Wenny D. L. Minimizing growth regulators in shoot cultures of an endangered plant, Hackelia venusta (Boraginaceae). Vitro Cell Dev. Biol. Plant 32: 267–271; 1996. doi:10.1007/BF02822699.
Fay M. F. Conservation of rare and endangered plants using in vitro methods. Vitro Cell Dev. Biol. Plant 28: 1–4; 1992. doi:10.1007/BF02632183.
Forte L.; Macchia F.; Scatigna A. M. E. Geological, pedological and phyto-climatic features of Lesina “Bosco Isola”. In: Macchia F. (ed) Biology and ecology of Cistus clusii Dunal. Editoriale Adda, Bari, Italy, pp 5–13; 2005. Reference translated from Italian language.
Giordano A. Pedology. Utet, Torino, Italy; 1999. Reference translated from Italian language.
Gonçalves S.; Romano A. Micropropagation of Drosophyllum lusitanicum (Dewy pine), an endangered West Mediterranean endemic insectivorous plant. Biodiversity Conserv. 14: 1071–1081; 2005. doi:10.1007/s10531-004-7846-z.
González-Benito M. E.; Martín C.; Iriondo J. M.; Pérez C. Conservation of the rare and endangered plants endemic to Spain. In: Benson E. E. (ed) Plant conservation biotechnology, Chapter 16. Taylor and Francis, London, UK, pp 251–264; 1999.
Hartmann H. T.; Kester D. E.; Davies F. T.; Geneve L. G. Hartmann and Kester’s plant propagation principles and pratices. Prentice Hall, Upper Saddle River, New Jersy, p 646; 2002.
Hernandez I.; Alegre L.; Munne-Bosch S. Drought-induced changes in flavonoids and other low molecular weight antioxidants in Cistus clusii grown under Mediterranean field conditions. Tree Physiol. 24: 1303–1311; 2004. doi:10.1093/treephys/24.11.1303.
Iriondo J. M.; Moreno C.; Perez C. Micropropagation of six rockrose (Cistus) species. Hort. Sci. 30(5): 1080–1081; 1995.
Iriondo J. M.; Perez C. Application of in vitro culture techniques to the conservation of Iberian endemic endangered plant species. Bot. Gard. Microprop. News 1: 4–6; 1990.
Leifert C.; Waites W. M.; Nicholas J. R. Bacterial contaminants of micropropagated plant cultures. J. Appl. Bacteriol. 67: 353–361; 1989.
Macchia F. Propagation and reproduction. In: Macchia F. (ed) Biology and ecology of Cistus clusii Dunal. Editoriale Adda, Bari, Italy, pp 43–48; 2005. Reference translated from Italian language.
Morone-Fortunato I.; Ruta C.; Castrignanò A.; Saccardo F. The effect of mycorrhizal symbiosis on the development of micropropagated artichokes. Sci. Hortic. 106: 472–483; 2005. doi:10.1016/j.scientia. 2005.05.006.
Murashige T.; Skoog R. A revised medium from rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15: 473–497; 1962.
Munnè-Bosch S.; Jubany-Marì T.; Alegre L. Enhanced photo- and antioxidative protection, and hydrogen peroxide accumulation in drought-stressed Cistus clusii and Cistus albidus plants. Tree Physiol. 23: 1–12; 2003. doi:10.1093/treephys/23.1.1.
Nitsch J. P.; Nitsch C. Haploid plants for pollen grains. Science 63: 85–87; 1969.
Pence V. C. The application of biotechnology for the conservation of endangered plants. In: Benson E. E. (ed) Plant conservation biotechnology, Chapter 15. Taylor and Francis, London, UK, pp 227–241; 1999.
Pignatti S. Italian flora. Edagricole, Bologna; 1982. Reference translated from Italian language.
Rech Filho, A.; Dal vesco, L. L.; Nodali, R. O.; Lischka, R. W.; Müller, C. V.; Guerra M. P. Tissue culture for the conservation and mass propagation of Vriesea reitzii Leme and Costa, a bromeliad threatened of extinction from the Brazilian Atlantic Forest. Biodiversity Conserv. 14: 1799–1808; 2005. doi:10.1007/s10531-004-0700-5.
Trigiano R. N.; Gray D. J. Plant tissue culture concepts and laboratory exercises. 2nd ed. CRC Press LLC, Boca Raton; 2000.
Wochok Z. S. The role of tissue culture in preserving threatened and endangered plant species. Biol. Conserv. 20: 83–89; 1981. doi:10.1016/0006-3207(81)90019-7.
Acknowledgments
The authors are grateful to Mr. Martin Baxter, English Language Professor at the Language Laboratory of Bari Polytechnic, for his valuable language assistance in the preparation of the manuscript. Thanks are due to Prof. Francesco Macchia, full professor of Botany and director of Botanic Gardens of Bari University, for providing C. clusii Dunal plants.
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Ruta, C., Morone-Fortunato, I. In vitro propagation of Cistus clusii Dunal, an endangered plant in Italy. In Vitro Cell.Dev.Biol.-Plant 46, 172–179 (2010). https://doi.org/10.1007/s11627-010-9284-9
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DOI: https://doi.org/10.1007/s11627-010-9284-9