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Shoot regeneration in stem expiants and its amenability to Agrobacterium tumefaciens mediated gene transfer in Brassica carinata

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Abstract

Immature stem segments of seven different genotypes of Brassica carinata produced shoots with variable frequencies when cultured in MS medium with BAP and picloram at 0.2 mg/l each. Line 171, which produced shoots with 100% efficiency from both cut ends of the expiant, was selected for testing the amenability of this regeneration protocol for genetic transformation. A non-oncogenic Agrobacterium tumefaciens containing plasmid PCV 730, a binary vector carrying resistance genes for kanamycin and hygromycin, was used. A cocultivation period of 4 d with a bacterial concentration of approximately 2.5×10 cells/ml, followed by a recovery period of 2 d, produced transformed shoots that could be selected and rooted in the presence of kanamycin at 15 mg/l. Transformation was confirmed by neomycin phospho-transferase assay and Southern blot analysis. Seed analysis of transformed plants indicated that kanamycin resistance was inherited in the progeny.

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Abbreviations

BAP:

6-Benzylaminopurine

Kn:

Kinetin

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

NAA:

∝-Naphthaleneacetic acid

ATP:

Adenosine triphosphate

MS:

Murashige and Skoog (1962) medium

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Authors and Affiliations

  1. Biotechnology Centre, Indian Agricultural Research Institute, 110012, New Delhi, India

    S. B. Narasimhulu, P. B. Kirti, T. Mohapatra, Shyam Prakash & V. L. Chopra

Authors
  1. S. B. Narasimhulu
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  2. P. B. Kirti
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  3. T. Mohapatra
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  4. Shyam Prakash
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  5. V. L. Chopra
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Communicated by G. C. Phillips

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Narasimhulu, S.B., Kirti, P.B., Mohapatra, T. et al. Shoot regeneration in stem expiants and its amenability to Agrobacterium tumefaciens mediated gene transfer in Brassica carinata . Plant Cell Reports 11, 359–362 (1992). https://doi.org/10.1007/BF00233366

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  • DOI: https://doi.org/10.1007/BF00233366

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