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Advances in Boerhaavia diffusa hairy root technology: a valuable pursuit for identifying strain sensitivity and up-scaling factors to refine metabolite yield and bioactivity potentials

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Abstract

The present study reports the Agrobacterium rhizogenes-mediated hairy root induction in of an ethno-medicinally significant herb–Boerhaavia diffusa L., for elucidating the underlying competence regarding its biosynthetic (i.e. boeravinone B and eupalitin) and bioactivity (antibacterial, antioxidant and anti-inflammatory) potentials. Host plant-specific receptiveness towards A. rhizogenes strains and disparity in compatibility threshold of leaf and nodal explants were evident. Only leaf explants responded, attaining hairy root induction with the ATCC 15834 followed by A4 and SA79 strains in reducing order of transformation efficiency. The growth behaviours differed amongst independent rhizoclones, and two clones of A4 (RBH) and ATCC 15834 (RBT8) origin demonstrated higher growth potentials. Polymerase chain reaction amplification of rol genes confirmed their transformed nature. Optimization of the appropriate solvent and reverse phase high-performance liquid chromatography parameters relating to the targeted metabolite production in the selected RBH and RBT8 clones revealed higher accumulation of eupalitin with the RBH clone having the best result of 1.44 times greater yield over the control root. Compared to the selected rhizoclones, the control roots however showed higher boeravinone B content. Devising a modified “stirred-tank” reactor through equipping with marine impellers and ring spargers facilitated high-density RBH root biomass yield with 6.1-fold and 1.15-fold yield increment of the boeravinone B and eupalitin respectively compared to shake-flask cultures. Considering the control roots, the RBH clone revealed analogous antioxidant/antibacterial activities with improved anti-inflammatory potential. The hairy root mediated higher production of boeravinone B and eupalitin could be achieved for the first time in bioreactor.

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Acknowledgments

The authors wish to express their sincere gratitude to the Director of CSIR-CIMAP, for providing the research facilities to carry out this work. Thanks are also due to the University Grants Commission (UGC), Department of Science & Technology (DST), Council of Scientific & Industrial Research (CSIR) and the Indian Council of Medical Research (ICMR)–New Delhi, India, for the financial support in the form of fellowships to RG, PP, SS, DKS and AS, respectively. Sincere appreciation is also due to the Academy of Scientific and Innovative Research, CSIR-CIMAP. The authors gratefully acknowledge the kind support from Dr. Prema Vasudeva. This work was supported by grants from the University Grants Commission (UGC–GAP 211).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Plant Biotechnology Department, Central Institute of Medicinal and Aromatic Plants (CSIR-CIMAP), Lucknow, 226015, Uttar Pradesh, India

    Ruby Gupta, Pallavi Pandey, Sailendra Singh & Suchitra Banerjee

  2. Academy of Scientific and Innovative Research, CSIR-CIMAP Campus, Lucknow, 226015, Uttar Pradesh, India

    Ruby Gupta, Pallavi Pandey & Sailendra Singh

  3. Molecular Bioprospection Division, Central Institute of Medicinal and Aromatic Plants (CSIR- CIMAP), Lucknow, 226015, Uttar Pradesh, India

    Dhananjay Kumar Singh, Archana Saxena, Suaib Luqman & Dnyaneshwar U. Bawankule

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  1. Ruby Gupta
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  2. Pallavi Pandey
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  3. Sailendra Singh
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Correspondence to Suchitra Banerjee.

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Gupta, R., Pandey, P., Singh, S. et al. Advances in Boerhaavia diffusa hairy root technology: a valuable pursuit for identifying strain sensitivity and up-scaling factors to refine metabolite yield and bioactivity potentials. Protoplasma 253, 1145–1158 (2016). https://doi.org/10.1007/s00709-015-0875-5

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