Abstract
An efficient protocol for hairy root induction in radish was established by optimizing several parameters that affect the efficiency of Agrobacterium rhizogenes-mediated transformations. Explants wounded using sterile hypodermic needle, infected with Agrobacterium suspension (0.6 OD600) for 10 min and co-cultivated in 1/2 MS medium containing acetosyringone (100 µM) for 2 days displayed maximum percentage of hairy root induction using MTCC 2364 (77.6%) and MTCC 532 (67.6%). On further experiments with MTCC 2364 initiated hairy roots, maximum biomass accumulation (fresh weight = 9.50 g; dry weight = 1.48 g) was achieved in liquid 1/2 MS medium supplemented with 87.6 mM sucrose after 40 days of culture. Transgenic state of hairy roots of MTCC 2364 was confirmed by polymerase chain reaction using rolB- and rolC-specific primers. The MTCC 2364-induced hairy roots produced higher amount of phenolic (33.0 mg g−1), flavonoid (48.0 mg g−1), and quercetin (114.8 mg g−1) content compared to auxin-induced roots of non-transformed radish. Furthermore, the results of ferric reducing antioxidant power and 1,1-diphenyl-2-picrylhydrazyl assay confirmed that the antioxidant activity of MTCC 2364 root extracts was improved when compared to auxin-induced roots of non-transformed radish. The present study offers a new insight in radish for production of phenolics and flavonoids (quercetin) using A. rhizogenes-mediated hairy root induction.
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Acknowledgements
Authors are thankful to the University Grant Commission: Basic Scientific Research (UGC-BSR Fellowship: No. UGC BSR-No.F.7-25/2007), Government of India, New Delhi for the financial assistance provided to M. Balasubramanian. Former Professor and current Vice-Chancellor, Bharathiar University, Coimbatore, Dr. A. Ganapathy is thanked for providing the strains MTCC 2364 and MTCC 532 of Agrobacterium rhizogenes. The authors would like to thank the Government of India for providing facilities through UGC-SAP (UGC-SAP-II:F-3-20/2013) and DST-FIST (DST-FIST:SR/FST/LSI-618/2014) Program.
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Girija designed research, supervised experiments at all the stages and performed critical revision of the manuscript. Balasubramanian performed all the experiments and analysed data. Anbumegala and Surendran assisted in conducting the experiments and helped in data interpretation. Arun and Balasubramanian wrote the manuscript and prepared figures. All authors read and approved the final manuscript.
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Balasubramanian, M., Anbumegala, M., Surendran, R. et al. Elite hairy roots of Raphanus sativus (L.) as a source of antioxidants and flavonoids. 3 Biotech 8, 128 (2018). https://doi.org/10.1007/s13205-018-1153-y
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DOI: https://doi.org/10.1007/s13205-018-1153-y