Abstract
Procedures were developed for rapid and prolific adventitious shoot regeneration of Arabidopsis thaliana (L.) Heynh ecotypes “Landsberg erect” and “C24” from cotyledon explants at 90–100% efficiency. Immature cotyledons had the highest shoot regeneration efficiency. Prolific regeneration was achieved in Murashige and Skoog medium (MS) supplemented with 0.1–0.4 mg−1 naphthalene acetic acid (NAA) and 1.0 mg−1 6-benzylaminopurine (BAP) within 2 weeks. The regenerated plants had a normal phenotype and produced fertile flowers and set seeds. The above regeneration protocol was used to develop a transformation method using disarmed strains of Agrobacterium tumefaciens strains pGV3850::pH1121 based on kanamycin (Km) selection. Transgenic shoots were produced within 2–3 weeks after inoculation. Transformation of shoots was confirmed by GUS histochemical assay, as well as southern blot hybridization.
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Abbreviations
- NAA:
-
1-Naphthalene acetic acid
- BAP:
-
6-Benzylaminopurine
- Km:
-
Kanamycin
- MSO:
-
Murashigea and Skoog medium [18] without hormonal supplements
- GUS:
-
<B>-Glucuronidase
- NB:
-
Nutrient broth
- NPT-II:
-
Neomycin phosphotransferase II
- X-Gluc:
-
5-Bromo-4-chloro-3-indolyl glucuronide
- CTAB:
-
Cetyl triethylammonium bromide
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Barghchi, M., Turgut, K., Scott, R. et al. High-frequency transformation from cultured cotyledons of Arabidopsis thaliana ecotypes “C24” and “Landsberg erecta”. Plant Growth Regul 14, 61–67 (1994). https://doi.org/10.1007/BF00024142
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DOI: https://doi.org/10.1007/BF00024142