Genetic transformation of Alstroemeria using particle bombardment

H.S. Lin, C. van der Toorn, C.J.J.M. Raemakers, R.G.F. Visser, M.J. de Jeu, E. Jacobsen

Research output: Contribution to journalArticleAcademicpeer-review

18 Citations (Scopus)

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 β-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.
Original languageEnglish
Pages (from-to)369-377
JournalMolecular Breeding
Volume6
Issue number4
DOIs
Publication statusPublished - 2000

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