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A simple and rapid Agrobacterium-mediated transformation protocol for cotton (Gossypium hirsutum L.): Embryogenic calli as a source to generate large numbers of transgenic plants

  • Genetic Transformation and Hybridization
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Abstract

A protocol is presented for efficient transformation and regeneration of cotton. Embryogenic calli co-cultivated with Agrobacterium carrying cry1Ia5 gene were cultured under dehydration stress and antibiotic selection for 3–6 weeks to generate several transgenic embryos. An average of 75 globular embryo clusters were observed on selection plates and these embryos were cultured on multiplication medium followed by development of cotyledonary embryos on embryo maturation medium to obtain an average of 12 plants per Petri plate of co-cultivated callus. About 83% of these plants have been confirmed to be transgenic by Southern blot analysis. An efficiency of ten kanamycin-resistant plants per Petri plate of co-cultivated embryogenic callus was obtained. The simplicity of the procedure and the efficiency of the initial material allow transformation of any variety where a single regenerating embryogenic callus line can be obtained. In addition, multiple transformations can be performed either simultaneously or sequentially. The method is extremely simple, reliable, efficient, and much less laborious than any other existing method for cotton transformation.

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Acknowledgement

We gratefully acknowledge the financial assistance of the Maharashtra State Seed Corporation Ltd.

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Authors and Affiliations

  1. International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, 110 067, New Delhi, India

    S. Leelavathi, V. G. Sunnichan, R. Kumria, G. P. Vijaykanth, R. K. Bhatnagar & V. S. Reddy

Authors
  1. S. Leelavathi
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  2. V. G. Sunnichan
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  3. R. Kumria
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  4. G. P. Vijaykanth
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  5. R. K. Bhatnagar
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  6. V. S. Reddy
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Corresponding author

Correspondence to S. Leelavathi.

Additional information

Communicated by H. Uchimiya

V.G. Sunnichan and R. Kumria contributed equally to this investigation

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Leelavathi, S., Sunnichan, V.G., Kumria, R. et al. A simple and rapid Agrobacterium-mediated transformation protocol for cotton (Gossypium hirsutum L.): Embryogenic calli as a source to generate large numbers of transgenic plants. Plant Cell Rep 22, 465–470 (2004). https://doi.org/10.1007/s00299-003-0710-x

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  • DOI: https://doi.org/10.1007/s00299-003-0710-x

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