Isolation of the lily embryo sac |
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Authors: | Vincent T Wagner Jouke P Kardolus Jac L Van Went |
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Institution: | (1) Department of Plant Cytology and Morphology, Agricultural University of Wageningen, Arboretumlaan 4, NL-6703 BD Wageningen, The Netherlands |
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Abstract: | Summary A 5%–8% yield of isolated embryo sacs of Lilium longiflorum was obtained using an enzymatic isolation procedure. The best results were obtained with a maceration mixture containing mannitol, pectinase, pectolyase, cellulase, hemicellulase, CaCl2 and NaOH. All developmental stages of the female gametophyte can be isolated in the living condition, although fewer than expected mature stages were observed. Moreover, only some of the more mature embryo sacs showed a positive fluorochromatic reaction (FCR) at the time of liberation. When the embryo sacs were stored in the enzyme or sugar solution after isolation, the positive reaction rapidly diminished for all stages. The isolated embryo sac and its nuclei were similar in size, shape, and position to the in situ embryo sac. Light microscopical observations of sectioned material revealed an intact cellular structure. However, the deleterious effects of the enzyme solution were sometimes observed in the form of lipid-like accumulations inside the isolated embryo sac.In collaboration with: Reconnaissance Cellulaire et Amélioration des Plantes, Université Cl. Bernard-Lyon I; INRA 879, 43, Boulevard du 11 Novembre 1918, F-69622 Villeurbanne Cedex, France |
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Keywords: | Embryo sac Lilium longiflorum Isolation technique Enzymatic maceration Fluorochromatic reaction |
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