Articles
AGROBACTERIUM-MEDIATED TRANSFORMATION OF PELARGONIUM (PELARGONIUM ZONALE HYBRIDS AND PELARGONIUM PELTATUM HYBRIDS)
Article number
725_103
Pages
737 – 746
Language
English
Abstract
Out of the genus Pelargonium two groups of hybrids are of great economic importance as bedding plants in Europe and North America, P. zonale (syn. P. x hortorum) and P. peltatum hybrids.
Both types are vegetatively propagated via cuttings.
A genetic transformation system is highly desired for these species.
The attempts of this investigation were to use a system for plant regeneration from explants of in vitro grown and greenhouse-grown plants for genetic transformation of important commercial Pelargonium zonale hybrid and P. peltatum hybrid cultivars.
To this aim, petiole explants were cultured on modified MS medium containing 10 µM TDZ. In eight out of sixteen tested cultivars, shoot regeneration has been obtained using explants from greenhouse-grown plants.
Explants from in vitro plantlets regenerated shoots in nine out of twelve investigated cultivars.
For selection of transgenic cells carrying the pat gene, a concentration of 2.5 µM glufosinate (syn. phosphinothricin) was shown to be appropriate.
Cefotaxim at 500 mg L-1 had no effect on shoot regeneration and did not inhibit the selective efficiency of glufosinate.
LBA4404 and EHA101 Agrobacterium strains carrying pIBGUS vector with pat gene as selectable marker gene and GUS gene as reporter gene were compared in transformation studies.
With regard to GUS expression in petiole explants sixteen days after coculture, better results were obtained with EHA 101 than with LBA 4404. The regeneration of GUS expressing shoots was demonstrated.
Transgenic shoots were able to build up a strong root system on glufosinate containing rooting medium.
By PCR the integration of the GUS gene was proven, while no DNA of the agrobacterial genome could be detected in the transgenic plantlets.
By these experiments, it was shown that pelargonium can be transformed via Agrobacterium tumefaciens using explants from adult plants.
Both types are vegetatively propagated via cuttings.
A genetic transformation system is highly desired for these species.
The attempts of this investigation were to use a system for plant regeneration from explants of in vitro grown and greenhouse-grown plants for genetic transformation of important commercial Pelargonium zonale hybrid and P. peltatum hybrid cultivars.
To this aim, petiole explants were cultured on modified MS medium containing 10 µM TDZ. In eight out of sixteen tested cultivars, shoot regeneration has been obtained using explants from greenhouse-grown plants.
Explants from in vitro plantlets regenerated shoots in nine out of twelve investigated cultivars.
For selection of transgenic cells carrying the pat gene, a concentration of 2.5 µM glufosinate (syn. phosphinothricin) was shown to be appropriate.
Cefotaxim at 500 mg L-1 had no effect on shoot regeneration and did not inhibit the selective efficiency of glufosinate.
LBA4404 and EHA101 Agrobacterium strains carrying pIBGUS vector with pat gene as selectable marker gene and GUS gene as reporter gene were compared in transformation studies.
With regard to GUS expression in petiole explants sixteen days after coculture, better results were obtained with EHA 101 than with LBA 4404. The regeneration of GUS expressing shoots was demonstrated.
Transgenic shoots were able to build up a strong root system on glufosinate containing rooting medium.
By PCR the integration of the GUS gene was proven, while no DNA of the agrobacterial genome could be detected in the transgenic plantlets.
By these experiments, it was shown that pelargonium can be transformed via Agrobacterium tumefaciens using explants from adult plants.
Authors
T. Winkelmann, K. Kayiani, M. Serek
Keywords
glufosinate, gene transfer, geranium, in vitro, phosphinothricin, regeneration, selection, thidiazuron
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