Genetic transformation of Alstroemeria using particle bombardment |
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Authors: | Lin Hsueh-Shih van der Toorn Caroline Raemakers Krit JJM Visser Richard GF De Jeu Marjo J Jacobsen Evert |
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Institution: | (1) Graduate School of Experimental Plant Sciences, Laboratorium of Plant Breeding, Wageningen Agricultural University, P.O. Box 386, Wageningen, Netherlands |
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Abstract: | Transgenic plants were obtained after particle bombardment of embryogenic callus derived from stem segments of two tetraploid Alstroemeria genotypes with plasmids containing different selection/reporter genes. Firstly, a plasmid containing a firefly luciferase reporter gene driven by the maize ubiquitin promoter (Ubi1), was bombarded into both friable embryogenic callus and proembryos. Transient and stable expression of luciferase was visually detected by a luminometer. This selection method is non-destructive and can be applied over the whole developmental process from callus to embryo and plantlet. Molecular proof of transformation was obtained both by PCR analysis and Southern hybridization. Secondly, a plasmid containing the bar gene together with an uidA gene coding for -glucuronidase both driven by the Ubi1 promoter was bombarded into proembryos. The transgenic callus was effectively selected from the callus clumps four months after bombardment on a medium containing 5 mg/l phosphinotricin (PPT). Selection by PPT was efficient and labour-saving. Stable expression of GUS was confirmed by the histochemical staining assay and molecular proof was obtained by PCR analysis. |
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Keywords: | Alstroemeria bar GUS luciferase ubiquitin promoter |
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