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Agrobacterium tumefaciens — mediated Transformation of Rice Using Coleoptile and Mature Seed-derived Callus

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Abstract

Morphologically normal, fertile transgenic rice plants (Oryza sativa L cv Taipei 309) were obtained using Agrobacterium tumefaciens strain LBA4404 harbouring the plasmid pTOK233. Two transgenic systems were developed. The first involved callus derived from mature seeds (scutellum) and, the second, used callus derived from 4-d-old coleoptiles. This is the first time that a coleoptile-based system has been used for producing transgenic rice plants. In the development of coleoptile based system, we have evaluated the effect of the length of callus induction period of the coleoptiles on transformation efficiency. The proportion of GUS positive plants was 23% in coleoptile experiment while in mature seed experiments it was 21%. Southern analyses were done to confirm the presence of the transgene. It was found that one to three copies of the transgene integrated in the transgenic plants.

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Abbreviations

AS:

acetosyringone

2,4-D:

2,4-dichlorophenoxyacetic acid

BAP:

6-benzylaminopurine

CB:

Carbenicillin

CF:

Cefotaxime

HYG:

hygromycin-B

Kn:

Kinetin

MES:

2-(N-morpholino)ethanesulphonic acid

MS:

Murashige and Skoog (1962)

NAA:

1-naphthaleneacetic acid

PGR:

Plant growth regulator

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Authors and Affiliations

  1. Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN, 47907, USA

    Tarun Kant, Halina Kononowicz-Hodges & Thomas K. Hodges

  2. Department of Botany, University of Rajasthan, Jaipur, 302 004, India

    Tarun Kant & S. L. Kothari

Authors
  1. Tarun Kant
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  2. S. L. Kothari
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  3. Halina Kononowicz-Hodges
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  4. Thomas K. Hodges
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Kant, T., Kothari, S.L., Kononowicz-Hodges, H. et al. Agrobacterium tumefaciens — mediated Transformation of Rice Using Coleoptile and Mature Seed-derived Callus. J. Plant Biochem. Biotechnol. 10, 121–126 (2001). https://doi.org/10.1007/BF03263120

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  • DOI: https://doi.org/10.1007/BF03263120

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