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Enhancement of genetic transformation efficiency in indica rice cultivar IR64 by vacuum infiltration and exogenous application of polyamines

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Abstract

Oryza sativa ssp. indica is not amenable to genetic modifications owing to its recalcitrant nature, rendering it difficult to transform and recover improved varieties to combat the persisting biotic and abiotic stress conditions. Therefore, in the present study, we have attempted to enhance transformation efficiency by applying polyamines and vacuum infiltration in Agrobacterium tumefaciens-mediated genetic transformation of Indica rice cv. IR64 seed-derived embryogenic calli. The highest number of regenerating calli, with maximum number of shoots per callus, was achieved in regeneration medium supplemented with 0.2 mM Spermidine, and the regenerated shoots displayed improved rooting when cultured on MS medium supplemented with 0.1 mM Putrescine. Further, vacuum infiltration of calli for 4 min resulted in the highest transformation efficiency of 8.1% as evident by GUS positive calli. The transformed calli were screened on the selection medium supplemented with hygromycin, and the integration and expression of the T-DNA in the rice genome were confirmed by GUS histochemical assay, PCR and Southern hybridization. The outcome of this study would be useful in micropropagation and genetic transformation studies of recalcitrant rice varieties.

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Acknowledgements

The authors acknowledge the funding support from ICAR-National Institute for Plant Biotechnology.

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Author notes

  1. Gadamchetty Pavan and Shruti Yadav have contributed equally.

Authors and Affiliations

  1. ICAR-National Institute for Plant Biotechnology, New Delhi, India

    Gadamchetty Pavan, Shruti Yadav & Suhas G. Karkute

  2. Deshbandhu College, University of Delhi, New Delhi, India

    Shruti Yadav & Kamana Singh

  3. Division of Crop Improvement, ICAR-Indian Institute of Vegetable Research, Varanasi, India

    Suhas G. Karkute

  4. Department of Biotechnology, Bharathidasan University, Tiruchirappalli, Tamil Nadu, India

    Markandan Manickavasagam

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  1. Gadamchetty Pavan
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  2. Shruti Yadav
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  3. Kamana Singh
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  4. Suhas G. Karkute
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  5. Markandan Manickavasagam
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Correspondence to Kamana Singh or Suhas G. Karkute.

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Communicated by Janusz Zimny.

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Pavan, G., Yadav, S., Singh, K. et al. Enhancement of genetic transformation efficiency in indica rice cultivar IR64 by vacuum infiltration and exogenous application of polyamines. CEREAL RESEARCH COMMUNICATIONS (2023). https://doi.org/10.1007/s42976-023-00455-6

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