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Plant Cell, Tissue and Organ Culture

, Volume 29, Issue 2, pp 101–108 | Cite as

Agrobacterium-mediated transformation and plant regeneration of buckwheat (Fagopyrum esculentum Moench.)

  • Jovanka Miljuš-Djukić
  • Mirjana Nešković
  • Slavica Ninković
  • Radomir Crkvenjakov
Article

Abstract

Genetic transformation of buckwheat (Fagopyrum esculentum Moench.) and regeneration of transgenic plants were obtained by using Agrobacterium tumefaciens strains as vectors. Buckwheat cotyledons were excised from imbibed seeds, co-cultivated with A. tumefaciens and subjected to previously reported protocols for callus and shoot regeneration. The transformation with oncogenic strains was confirmed by opine and DNA analyses of tumour tissue extracts. Plants were regenerated on cotyledon fragments incubated with strain A281, harboring pGA472, which carries the neomycin phosphotransferase II gene for kanamycin resistance. The transformation of resistant shoot clones was confirmed by NPTII enzyme assay and DNA hybridization. A large number of transformed shoots were rooted and fertile plantlets were raised in the greenhouse. Transgenic plants comprised pin and thrum clones, which were allowed to cross-pollinate. In about 180 R2 seeds tested for kanamycin resistance, the ratio of resistant to sensitive seedlings was roughly 3:1.

Key words

Agrobacterium tumefaciens buckwheat Fagopyrum esculentum NPTII gene transgenic plants 

Abbreviations

BAP

6-benzylaminopurine

2,4-D

dichloro-phenoxyacetic acid

2iP

6-(γ, γ,-dimethylallyl-amino)-purine

IBA

indole-3-butyric acid

IAA

indole-3-acetic acid

Km

kanamycin

NPTII

neomycin phosphotransferase II

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Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • Jovanka Miljuš-Djukić
    • 1
  • Mirjana Nešković
    • 1
    • 2
  • Slavica Ninković
    • 1
  • Radomir Crkvenjakov
    • 3
  1. 1.Institute for Biological Research ‘S. Stanković’University of BelgradeBelgradeYugoslavia
  2. 2.Institute of Botany, Faculty of ScienceUniversity of BelgradeBelgradeYugoslavia
  3. 3.Institute for Molecular Biology and Genetic EngineeringUniversity of BelgradeBelgradeYugoslavia

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