Regeneration of fertile plants from protoplasts of different Arabidopsis thaliana genotypes |
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Authors: | Brigitte Damm and Lothar Willmitzer |
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Institution: | (1) Institut für Genbiologische Forschung Berlin GmbH, Ihnestraße 63, D-1000 Berlin 33 |
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Abstract: | Summary A protocol for obtaining regenerated fertile plants from mesophyll protoplasts of Arabidopsis thaliana is reported. Protoplasts were isolated from leaves of 21-to 28-day-old Arabidopsis plants grown in a controlled environment. Sustained divisions were achieved when protoplasts were embedded in beads formed by 1.4% sodium alginate in the presence of 50mM CaCl2 in 0.4 mannitol, which was then exchanged againts modified B5 medium. About 0.4%–0.6% of the protoplasts developed into colonies of which 80%–90% formed shoots and subsequently regenerated to fertile plants. Seeds harvested from more than 200 independently regenerated plants were sown and germination frequencies of more than 95% were obtained. Furthermore, the F1 plants did not show any evidence of somaclonal variation on visual inspection. This protocol was originally developed for Arabidopsis thaliana Columbia ; however it was shown to be applicable also for the genotypes Wassilewskija , Landsberg erecta and Estland though with differing efficiencies.Abbreviations FDA
fluorescein diacetate
- CM
culture medium
- SRM
shoot regeneration medium
- SEM
shoot elongation medium
- RM
rooting medium
- PE
plating effciency
- NAA
-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- BAP
6-benzylaminopurine
- Kin
kinetin
- 2-iP
2-isopentenyladenine
- GA3
gibberetic acid |
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Keywords: | Arabidopsis thaliana Protoplast Plant regeneration Alginate |
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