Articles
DEVELOPMENT OF A PLANT REGENERATION SYSTEM APPLICABLE FOR GENE TRANSFORMATION IN THE ORNAMENTAL ALSTROEMERIA
Article number
508_7
Pages
61 – 68
Language
Abstract
Development of a plant regeneration system is a prerequisite for genetic transformation.
Two plant regeneration systems, based on either leaf explants or compact calli, were developed in Alstroemeria and were used for particle bombardment-mediated transformation.
Gold particles coated with two different plasmids’ DNA were separately bombarded into the target material.
One plasmid contained the luc gene coding for firefly luciferase (LUC), and the other one contained the UidA gene coding for
-glucuronidase (GUS). All these genes were driven by the maize ubiquitin promoter (Ubil). In the present study the expression of the reporter genes LUC and GUS was investigated.
The leaf explants directly produced adventitious shoots, and the compact calli formed plants indirectly via embryogenesis.
Luciferase activity was observed in 28.9% of the bombarded leaf explants 5 days after bombardment.
GUS expression was observed in 59.7% of the bombarded leaf explants 4 weeks after bombardment.
Both genes are transiently expressed.
On the other hand, 1% and 3.1% of the bombarded compact calli regenerated plants, respectively, with stable expression of luciferase and GUS genes.
All those transgenic plants were shown to be chimeric.
The compact callus shown to be superior to the leaf explants in particle bombardment-mediated transformation.
Two plant regeneration systems, based on either leaf explants or compact calli, were developed in Alstroemeria and were used for particle bombardment-mediated transformation.
Gold particles coated with two different plasmids’ DNA were separately bombarded into the target material.
One plasmid contained the luc gene coding for firefly luciferase (LUC), and the other one contained the UidA gene coding for
-glucuronidase (GUS). All these genes were driven by the maize ubiquitin promoter (Ubil). In the present study the expression of the reporter genes LUC and GUS was investigated.The leaf explants directly produced adventitious shoots, and the compact calli formed plants indirectly via embryogenesis.
Luciferase activity was observed in 28.9% of the bombarded leaf explants 5 days after bombardment.
GUS expression was observed in 59.7% of the bombarded leaf explants 4 weeks after bombardment.
Both genes are transiently expressed.
On the other hand, 1% and 3.1% of the bombarded compact calli regenerated plants, respectively, with stable expression of luciferase and GUS genes.
All those transgenic plants were shown to be chimeric.
The compact callus shown to be superior to the leaf explants in particle bombardment-mediated transformation.
Authors
H.S. Lin, M.J. De Jeu, E. Jacobsen
Keywords
Alstroemeria, callus, leaf culture, particle bombardment, transformation
Online Articles (68)