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Meta-topolin enhances regeneration and Agrobacterium-mediated genetic transformation in radish (Raphanus sativus L.)

  • Plant Tissue Culture
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Abstract

Meta-topolin (mT) is a novel aromatic cytokinin that stimulates morphogenesis and is an alternative source of cytokinins frequently employed in regeneration systems. Hence, the present research explored the prospect of mT for improving regeneration and genetic transformation efficiency. Cotyledonary node explants were cultured on optimum plant growth regulator medium incorporated with mT for enhanced shoot induction (93.6%, 2.0 mg l−1 BA and 0.9 mg l−1 mT), elongation (89.3%, 2.0 mg l−1 GA3 and 0.9 mg l−1 mT), and rooting (90.3%, 0.9 mg l−1 IBA and 0.9 mg l−1 mT) respectively. The plant transformation study was carried out through Agrobacterium-mediated transformation using the pCAMBIA1301 vector containing the LB4404 strain for standardizing transformation strategies. Transformed shoots and rooting were determined in two stages using 10 and 6 mg l−1 hygromycin B for exterminating chimeric explants. After co-cultivation, explants were cultured at the optimal concentration of 0.9 mg l−1 mT, 2.0 mg l−1 BA, 2.0 mg l−1 GA3, and 0.9 mg l−1 IBA for enhanced transformation efficiency (27.3%), corresponding to transformed regeneration without mT (18.6%) occurring with less efficiency. The existence of transgenes in the radish genome was ascertained by the GUS assay, PCR, RT-PCR, and qRT-PCR. Overall, our investigation demonstrated that including mT increases regeneration and enhances transformation efficiency in radish. Therefore, diverse radish varieties could use a designed transformation strategy to acquire essential traits.

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Acknowledgements

Dr. Sivabalan Karthik expresses his gratitude to the Jawaharlal Nehru Memorial Fund, New Delhi, India, for providing a Jawaharlal Nehru Scholarship (Ref no: SU-1/88/2016-17/79) to support this research. Furthermore, this research has been partially supported by Kangwon National University Post-Doctoral Support Program No-2021038. The first author sincerely thanks Prof. Dr. Hyeran Kim, Department of Biological Sciences, Kangwon National University, South Korea, for inspiring and sustaining the research work.

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Authors and Affiliations

  1. Department of Biological Sciences, Kangwon National University, Chuncheon, 24341, South Korea

    Sivabalan Karthik

  2. Department of Biotechnology, Bharathidasan University, Tamil Nadu, Tiruchirappalli, 620 024, India

    Sivabalan Karthik, Selvam Sathish & Markandan Manickavasagam

  3. Department of Bioinformatics, Alagappa University, Tamil Nadu, Karaikudi, 630 003, India

    Jesudass Joseph Sahayarayan

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  1. Sivabalan Karthik
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Correspondence to Markandan Manickavasagam.

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Karthik, S., Sathish, S., Sahayarayan, J.J. et al. Meta-topolin enhances regeneration and Agrobacterium-mediated genetic transformation in radish (Raphanus sativus L.). In Vitro Cell.Dev.Biol.-Plant 58, 806–815 (2022). https://doi.org/10.1007/s11627-022-10311-7

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