Agrobacterium rhizogenes-mediated transformation and plant regeneration of four Gentiana species |
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Authors: | Ivana Momčilović Dragoljub Grubišić Milan Kojić Mirjana Nešković |
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Institution: | (1) lnstitute for Biological Research “S. Stanković”, University of Belgrade, 29 novembra 142, 11060 Belgrade, Yugoslavia;(2) Institute for Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Yugoslavia |
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Abstract: | Shoots of micropropagated Gentiana acaulis, G. cruciata, G. lutea, and G. purpurea were inoculated with suspensions of Agrobacterium
rhizogenes cells, strains ATCC 15834 or A4M70GUS. Adventitious roots appeared at the sites of inoculation in all 4 species.
Root tips were excised and cultured on growth regulator-free media for 2-6 years. They exhibited very high branching and plagiotropism.
Spontaneous bud initiation occurred in roots of G. cruciata. Roots of G. lutea, G. acaulis and G. purpurea were cultured on
media with high kinetin concentration, which induced the formation of friable callus tissues. Only in G. purpurea were these
calluses organogenic. Regenerated shoots of G. cruciata and G. purpurea gave rise to plants, that displayed the typical phenotypes
of A. rhizogenes-transformed plants: short internodes and rolled leaves. In the roots of G. acaulis and G. cruciata, transformed
with A. rhizogenes A4M70GUS, a positive reaction with X-gluc indicated the activity of β-glucuronidase. The DNA extracted
from hairy roots and from the roots of transgenic plants hybridized with the appropriate genomic probes in Southern blotting.
This is taken as evidence of the stable genetic transformation in the 4 Gentiana species.
This revised version was published online in June 2006 with corrections to the Cover Date. |
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Keywords: | Agrobacterium rhizogenes Gentiana acaulis L G cruciata L G lutea L G purpurea L hairy roots plant regeneration |
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