Articles
TRANSFORMATION OF GENTIAN WITH AGROBACTERIUM RHIZOGENES
Article number
392_18
Pages
153 – 160
Language
Abstract
Hairy roots of gentian (Gentiana scabra Bunge var. buergeri Maxim.). were induced following the inoculation with wild type of Agrobacterium rhizogenes strain MAFF03–01724. These hairy roots, after the elimination of bacteria, vigorously grew in hormone free ½ MS liquid medium (half strength of Murashige and Skoog (Murashige and Skoog, 1962) medium, 1.5% sucrose), at 25°C in the dark (at 100 rpm). Adventitious shoots were differentiated from the hairy roots by culturing on ½ MS solid medium containing 2–3 mg/1 6-benzylaminopurine (BAP) and 0.1–0.2 mg/1 1-naphthaleneacetic acid (NAA) at 25°C under 16 h/day light.
Then the whole plants were regenerated by transfer to hormone free MS solid medium.
The integration of Ri plasmid T-DNA was confirmed by detection of mikimopine (opine) synthesis in the hairy roots.
The transformed plants, when the regenerated plants were cultured on hormone free MS solid medium, showed elongation of internodes and vigorous emergence of lateral branches at the base of the stems, while the non-transformed plants showed rosette phenotype.
Furthermore, in comparison with the non-transformed plants, the transformed plants showed several morphologic alterations such as wrinkled leaves, reduced apical dominance, increased root growth, and plagiotropic roots.
About 30 days after the transfer to the soil in the pots, the transformed plants formed flower buds, while the non-transformed plants still kept showing rosette phenotype.
Then the whole plants were regenerated by transfer to hormone free MS solid medium.
The integration of Ri plasmid T-DNA was confirmed by detection of mikimopine (opine) synthesis in the hairy roots.
The transformed plants, when the regenerated plants were cultured on hormone free MS solid medium, showed elongation of internodes and vigorous emergence of lateral branches at the base of the stems, while the non-transformed plants showed rosette phenotype.
Furthermore, in comparison with the non-transformed plants, the transformed plants showed several morphologic alterations such as wrinkled leaves, reduced apical dominance, increased root growth, and plagiotropic roots.
About 30 days after the transfer to the soil in the pots, the transformed plants formed flower buds, while the non-transformed plants still kept showing rosette phenotype.
Authors
C. Suginuma, T. Akihama
Keywords
Gentian(Gentiana scabra Bunge var. buergeri Maxim.), Agrobacterium rhizogenes, Transformation, Hairy root, Regeneration
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