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High-frequency transformation from cultured cotyledons of Arabidopsis thaliana ecotypes “C24” and “Landsberg erecta

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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Correspondence to Mehdi Barghchi.

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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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Key words

  • Arabidopsis
  • regeneration
  • cotyledon
  • transgenic plants
  • antibiotics