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A simple and efficient Agrobacterium-mediated procedure for transformation of tomato

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Abstract

We describe a highly efficient and reproducible Agrobacterium-mediated transformation protocol applicable to several varieties of tomato (Solanum lycopersicum, earlier known as Lycopersicum esculentum). Conditions such as co-cultivation period, bacterial concentration, concentration of benzyl amino purine (BAP), zeatin and indole acetic acid (IAA) were optimized. Co-cultivation of explants with a bacterial concentration of 108 cells/ml for three days on 2 mg/l BAP, followed by regeneration on a medium containing 1 mg/ml zeatin resulted in a transformation frequency of 41.4%. Transformation of tomato plants was confirmed by Southern blot analysis and β-glucuronidase (GUS) assay. The protocol developed showed very high efficiency of transformation for tomato varieties Pusa Ruby, Arka Vikas and Sioux. The optimized transformation procedure is simple, efficient and does not require tobacco, Petunia, tomato suspension feeder layer or acetosyringone.

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Abbreviations

AMV:

alfalfa mosaic virus

BAP:

6-benzylaminopurine

DMSO:

dimethyl sulphoxide

GUS:

β-glucuronidase

IAA:

indole-3-acetic acid

MS:

Murashige and Skoog

OD:

optical density

PCR:

polymerase chain reaction

SDS:

sodium dodecyl sulphate

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Authors and Affiliations

  1. Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110 021, India

    Manoj K. Sharma, Amolkumar U. Solanke, Dewal Jani & Arun K. Sharma

  2. Institute of Genomics and Integrative Biology, Delhi, 110 007, India

    Yogendra Singh

Authors
  1. Manoj K. Sharma
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  2. Amolkumar U. Solanke
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  3. Dewal Jani
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  4. Yogendra Singh
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  5. Arun K. Sharma
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Correspondence to Arun K. Sharma.

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Sharma, M.K., Solanke, A.U., Jani, D. et al. A simple and efficient Agrobacterium-mediated procedure for transformation of tomato. J Biosci 34, 423–433 (2009). https://doi.org/10.1007/s12038-009-0049-8

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  • DOI: https://doi.org/10.1007/s12038-009-0049-8

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