Stable transformation and long-term expression of the gusA reporter gene in callus lines of perennial ryegrass (Lolium perenne L.)

Stable transformation of perennial ryegrass (Lolium perenne L.) was achieved by biolistic bombardment of a non embryogenic cell suspension culture, using the hpt and gusA gene. The transformation yielded on the average 5 callus lines per bombardment (1.4 x 10(6) cells). Stable integration of the gen...

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Bibliographic Details
Published in:Plant molecular biology 1994, Vol.24 (2), p.401-405
Main Authors: VAN DER MAAS, H. M, DE JONG, E. R, RUEB, S, HENSGENS, L. A. M, KRENS, F. A
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Blotting, Southern
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation
Genes, Reporter
Genetic engineering
Genetic technics
Glucuronidase - genetics
Glucuronidase - metabolism
Lolium - genetics
Methods. Procedures. Technologies
Transformation, Genetic
Transgenic animals and transgenic plants
Transgenic plants
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Summary:Stable transformation of perennial ryegrass (Lolium perenne L.) was achieved by biolistic bombardment of a non embryogenic cell suspension culture, using the hpt and gusA gene. The transformation yielded on the average 5 callus lines per bombardment (1.4 x 10(6) cells). Stable integration of the genes into the plant genome was demonstrated by Southern analysis of DNA, isolated from hygromycin-resistant callus lines. The gusA reporter gene, which was regulated by the constitutive promoter of the rice gene GOS2, was expressed in both transient and stable transformation assays, indicating that this promoter is suitable for expression of a transferred gene in perennial ryegrass. Long-term GUS expression was observed in ca. 40% of the callus lines, whereas the other callus lines showed instability after 6 months and 1 year of culture.
ISSN:0167-4412
1573-5028
DOI:10.1007/BF00020178