Abstract
A protocol for somatic embryogenesis and plant regeneration of Ilexparaguariensis St. Hil. from embryos cultures was developed. Heart stage zygotic embryos were removed from seeds of immature, light green fruit and treated with antimicrotubule agents (0.1; 0.2, and 0.5% colchicine for 24 and 48 h; 1; 10, and 20 μM of either trifluralin, ααα- trifluoro- 2,6-dinitro-N,N- dipropyl-p-toluidine, or oryzalin, 3,5-dinitro-N4, N-dipropylsulphate during 48 h). The embryos were cultured aseptically on quarter-strength Murashige and Skoog medium containing 3% sucrose, 0.65% agar (1/4MS), and 0.46 μM zeatin. Cultures were incubated in darkness at 27 ± 2 °C. All thetreatments provoked a diminution of the number of germinated embryos and in some of the treated embryos somatic embryogenesis was induced. Somatic embryo maturation and conversion into whole plants could be achieved by culturing the embryos on 1/4MS lacking hormones and incubated at 27 ± 2 °C, 14 h photoperiod (116 μmol m-2s-1). Mostof the plants regenerated from somatic embryos appeared morphologically normaland grew under greenhouse conditions. Only 2 plants out of 152 studied contained the tetraploid number of the chromosomes (2n = 4x = 80), meanwhile the rest of the plants had the normal diploid number of chromosomes (2n =2x = 40). Somatic embryos with abnormal morphology were also observed.
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References
Alemanno, L. & E. Guiderdoni, 1994. Increased doubled haploid plant regeneration from rice (Oryza sativa L.) anthers cultured on colchicine-supplemented media. Plant Cell Rep 13: 432–436.
Ammirato, P.V., 1989. Recent progress in somatic embryogenesis. IAPTC Newsl 57: 1–16.
Bartels, P.G. & J.L. Hilton, 1973. Comparison of trifluralin, oryzalin, pronamide, propham and colchicine treatments on microtubules. Pest Bioch Physiol 3: 462–472.
Bucheim, J., S. Colburn & J. Ranch, 1989. Maturation of soybean somatic embryos and the transition to plantlets growth. Plant Physiol 89: 768–775.
Dunstan, D.I., T.E. Tautorus & T.A. Thorpe, 1995. Somatic embryogenesis in woody plants. In: T.A. Thorpe (Ed.), In Vitro Embryogenesis in Plants, pp. 471–538. Kluwer Academic Publishers, Dordrecht.
Eigsti, D.I. & P. Dustin, 1955. Spindle and cytoplasm. Colchicine. In: Agriculture, pp. 65–139. The Iowa State College Press, Ames.
Giberti, G.C., 1995. Ilex en Sudamérica: Florística, Sistemática y potencialidades con relación a un banco de germoplasma para la yerba mate. In: Erva-mate: Biología e cultura no cono sul, pp. 303–312. UFRGS, Brasil.
Hansen, N.J.P. & S.B. Andersen, 1998a. In vitro chromosome doubling with colchicine during microspore culture in wheat (Triticum aestivum L.). Euphytica 102: 101–108.
Hansen, N.J.P. & S.B. Andersen, 1998b. Efficient production of doubled haploid wheat plants by in vitro treatment of microspores with trifluralin or APM. Euphytica 117: 401–405.
Hansen, A.L., A. Gertz, M. Joersbo & S.B. Andersen, 1998. Antimicrotubule herbicides for in vitro chromosome doubling in Beta vulgaris L. ovule culture. Euphytica 101: 231–237.
Hu, C.Y. & I.M. Sussex, 1971. In vitro development of embryoids on cotyledons of Ilex aquifolium. Phytomorphology 21: 103–107.
Hu, C.Y., 1989. Holly (Ilex spp.). In: Y.P.S. Bajaj (Ed.) Biotechnology in Agriculture and Forestry, Vol 5, pp. 412–427. Springer-Verlag, Berlin.
Hu, C.Y., J.D. Ochs & F.M. Mancini, 1978. Further observations on Ilex embryoid production. Z Pflanzenphysiol 89: 41–49.
Hugdahl, J.D. & L.C. Morejohn, 1993. Rapid and reversible highaffinity binding of the dinitroaniline herbicide oryzalin to tubulin from Zea mays L. Plant Physiol 102: 725–740.
Hunault, G., 1997. Tétraploidisation du fenouil (Foeniculum vulgare Miller) par application de colchicine à une suspension cellulaire embryogène. Acta bot Gallica 144: 83–93.
Iqbal, M.C.M., C. Möllers & G. Röbbelen, 1994. Increased embryogenesis after colchicine treatment of microspore cultures of Brassica napus L. J Plant Physiol 143: 222–226.
Konar, R.N. & Y.P. Oberoi, 1965. In vitro development of embryoids on the cotyledons of Biota orientalis. Phytomorphology 15: 137–140.
Linsmaier, E.M. & F. Skoog, 1965. Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18: 100–127.
Michler, C.H. & E.O. Bauer, 1991. High frequency somatic embryogenesis from leaf tissue of Populus spp. Plant Sci 77: 111–118.
Monnier, M., 1995. Culture of zygotic embryos. In: T.A. Thorpe (Ed.), In Vitro Embryogenesis in Plants, pp. 117–153. Kluwer Academic Publishers, Dordrecht.
Murashige, T. & F. Skoog, 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497.
Redha, A., T. Attia, B. Büter, S. Saisingtong, P. Stamp & J.E. Schmid, 1998. Improved production of doubled haploids by colchicine application to wheat (Triticum aestivum L.) anther culture. Plant Cell Rep 17: 974–979.
Rey, H.Y. & L.A. Mroginski, 1988. Regeneración de plantas de yerba mate (Ilex paraguariensis) por cultivo in vitro de ápices caulinares y de segmentos nodales. Phyton 48: 139–145.
Sansberro, P.A., H.Y. Rey, L.A. Mroginski & M.M. Collavino, 1998. In vitro culture of rudimentary embryos of Ilex paraguariensis: Responses to exogenous cytokinins. J Plant Growth Reg 17: 101–105.
Sansberro, P.A, H.Y. Rey, L.A. Mroginski & M.M. Collavino, 1999. In vitro plant regeneration of Ilex paraguariensis (Aquifoliaceae). In Vitro Cell Dev Biol – Plant 35: 401–402.
Sefc, K.M., P. Ruckenbauer & F. Regner, 1997. Embryogenesis in microspore culture of Vitis subspecies. Vitis 36: 15–20.
Sharma, D.R., R. Kaur & K. Kumar, 1996. Embryo rescue in plants – A review. Euphytica 89: 325–337.
Song, P., Kang & E.B. Peffley, 1997. Chromosome doubling of Allium fistulosum × A. cepa interspecific F1 hybrids through colchicine treatment of regenerating callus. Euphytica 93: 257–262.
Tisserat, B., E.B. Esan & T. Murashige, 1979. Somatic embryogenesis in angiosperms. Horticultural Rev 1: 1–78.
Wetzstein, H.Y. & C.C. Baker, 1993. The relationship between somatic embryo morphology and conversion in peanut (Arachis hypogaea L.). Plant Sci 92: 81–89.
Zaki, M.A. & H. Dickinson, 1991. Microspore-derived embryos in Brassica: the significance of division symmetry in pollen mitosis I to embryogenic development. Sex Plant Rep 4: 48–55.
Zaki, M.A. & H.G. Dickinson, 1995. Modification of cell development in vitro: The effect of colchicine on anther and isolated microspore culture in Brassica napus. Plant Cell Tissue Org Cult 40: 255–270.
Zhao, J., D.H. Simmonds & W. Newcomb, 1996. High frequency production of doubled haploid plants of Brassica napus cv. Topas derived from colchicine-induced microspore embryogenesis without heat shock. Plant Cell Rep 15: 668–671.
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Rey, H., Sansberro, P., Collavino, M. et al. Colchicine, trifluralin, and oryzalin promoted development of somatic embryos in Ilex paraguariensis (Aquifoliaceae) . Euphytica 123, 49–56 (2002). https://doi.org/10.1023/A:1014489207888
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DOI: https://doi.org/10.1023/A:1014489207888