Articles
THE EFFECT OF HYGROMYCIN ON REGENERATION IN DIFFERENT ALSTROEMERIA EXPLANT TYPES AFTER AGROBACTERIUM-MEDIATED TRANSFORMATION
Article number
572_11
Pages
97 – 103
Language
English
Abstract
This is the first successful report of Agrobacterium-mediated transformation in Alstroemeria by infection of FEC (Friable Embryogenic Callus) lines and leaves with axil tissues.
Of the transformation methods, particle bombardment and Agrobacterium-mediated transformation have been widely used to transfer foreign DNA into the plant genome.
Especially, Agrobacterium tumefaciens efficiently infects most plants.
Most monocotyledonous plants, including Alstroemeria, are recalcitrant to A. tumefaciens infection.
Therefore, it is essential to develop an efficient, reliable protocol for Agrobacterium-mediated transformation in Alstroemeria. In order to achieve this aim, different hygromycin concentrations in selection medium were tested and compared in FEC explants and leaves with axil tissues using Agrobacterium strain LBA4404 (pTOK233). As a result, 20 mg/l hygromycin was a proper concentration to select for both FEC lines and leaves with axil tissues.
Using this concentration, transformed lines were obtained.
Moreover, FEC lines showed at least 2.5 times higher rate in the survival rate, and 5 times higher rate in transient GUS gene expression than those of leaves with axil tissues.
The protocol, seems to be a promising method for the transformation of the monocot Alstroemeria with genes of interest in the near future.
Of the transformation methods, particle bombardment and Agrobacterium-mediated transformation have been widely used to transfer foreign DNA into the plant genome.
Especially, Agrobacterium tumefaciens efficiently infects most plants.
Most monocotyledonous plants, including Alstroemeria, are recalcitrant to A. tumefaciens infection.
Therefore, it is essential to develop an efficient, reliable protocol for Agrobacterium-mediated transformation in Alstroemeria. In order to achieve this aim, different hygromycin concentrations in selection medium were tested and compared in FEC explants and leaves with axil tissues using Agrobacterium strain LBA4404 (pTOK233). As a result, 20 mg/l hygromycin was a proper concentration to select for both FEC lines and leaves with axil tissues.
Using this concentration, transformed lines were obtained.
Moreover, FEC lines showed at least 2.5 times higher rate in the survival rate, and 5 times higher rate in transient GUS gene expression than those of leaves with axil tissues.
The protocol, seems to be a promising method for the transformation of the monocot Alstroemeria with genes of interest in the near future.
Publication
Authors
J.B. Kim, M.J. de Jeu, C.J.J.M. Raemakers, E. Jacobsen, R.G.F. Visser
Keywords
friable embryogenic callus, GUS, leaves with axils, monocot, organogenic tissue, somatic embryo
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