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Agrobacterium rhizogenes-mediated transformation of Picrorhiza kurroa Royle ex Benth.: establishment and selection of superior hairy root clone

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Abstract

A protocol for induction and establishment of Agrobacterium rhizogenes-mediated hairy root cultures of Picrorhiza kurroa was developed through optimization of the explant type and the most suitable bacterial strain. The infection of leaf explants with the LBA9402 strain resulted in the emergence of hairy roots at 66.7% relative transformation frequency. Nine independent, opine and TL-positive hairy root clones were studied for their growth and specific glycoside (i.e., kutkoside and picroside I) productivities at different growth phases. Biosynthetic potentials for the commercially desirable active constituents have been expressed by all the tested hairy root clones, although distinct inter-clonal variations could be noted in terms of their quantity. The yield potentials of the 14-P clone, both in terms of biomass as well as individual glycoside contents (i.e., kutkoside and picroside I), superseded that of all other hairy root clones along with the non-transformed, in vitro-grown control roots of P. kurroa. The present communication reports the first successful establishment, maintenance, growth and selection of superior hairy root clone of Picrorhiza kurroa with desired phyto-molecule production potential, which can serve as an effective substitute to its roots and thereby prevent the indiscriminate up-rooting and exploitation of this commercially important, endangered medicinal plant species.

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Abbreviations

BAP:

benzyl amino purine

IBA:

indole-3-butyric acid

NAA:

α-naphthalene acetic acid

TL:

Left-terminus DNA

References

  • Anonymous (2001) Picrorhiza kurroa Monograph. Altern Med Rev 6:319–321

  • Banerjee S, Zehra M, Kukreja AK, Kumar S (1995) Hairy roots in medicinal plants. Curr Res Med Arom Plants 17:348–378

    Google Scholar 

  • Byrne M, Mc Donnell R, Wright M, Carnes M (1987) Strain and genotype specificity in Agrobacterium soybean interaction. Plant Cell Tissue Organ Cult 8:3–5

    Article  CAS  Google Scholar 

  • Christey MC, Braun RH (2005) Production of hairy root cultures and transgenic plants by Agrobacterium rhizogenes-mediated transformation. Methods Mol Biol 286:47–60

    PubMed  CAS  Google Scholar 

  • Constabel CP, Towers GHN (1988) Thiarubine accumulation in hairy root cultures of Chaenactis douglasii. J Plant Physiol 133:67–72

    CAS  Google Scholar 

  • Dubey NK, Kumar R, Tripathi P (2004) Global promotion of herbal medicine: India’s opportunity. Curr Sci 86:37–41

    Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  PubMed  CAS  Google Scholar 

  • Giri A, Ravindra ST, Dhingra V, Narasu ML (2001) Influence of different strains of Agrobacterium rhizogenes on induction of hairy roots and artemisinin production in Artemisia annua. Curr Sci 81:378–382

    CAS  Google Scholar 

  • Guillon S, Tremouillaux-Guiller J, Pati PK, Rideau M, Gantet P (2006a) Harnessing the potential of hairy roots: dawn of a new era. Trends Biotechnol 24:403–409

    Article  PubMed  CAS  Google Scholar 

  • Guillon S, Tremouillaux-Guiller J, Pati PK, Rideau M, Gantet P (2006b) Hairy root research: recent scenario and exciting prospects. Curr Opin Plant Biol 9:341–346

    Article  PubMed  CAS  Google Scholar 

  • Gupta PP (2001) Picroliv. Drugs Future 26:25–31

    CAS  Google Scholar 

  • Gupta A, Khajuria A, Singh J, Bedi KL, Satti NK, Dutt P, Suri KA, Suri OP, Qazi GN (2006) Immunomodulatory activity of biopolymeric fraction RLJ-NE-205 from Picrorhiza kurroa. Int Immunopharmacol 6:1543–1549

    Article  PubMed  CAS  Google Scholar 

  • Hamil JD, Parr AJ, Robins RJ, Rhodes MJC (1986) Secondary product formation by cultures of Beta vulgaris and Nicotiana rustica transformed with Agrobacterium rhizogenes. Plant Cell Rep 5:111–114

    Article  Google Scholar 

  • Hooykass PJJ, Klapwijk PM, Nuti PM, Shilperoot RA, Rorsch A (1977) Transfer of the Agrobacterium tumefaciens Ti-plasmid to a virulent Agrobacteria and Rhizobium explanta. J Gen Microbiol 98:477–487

    Google Scholar 

  • Hu Zhi-Bi, Du Min (2006) Hairy roots and its application in plant genetic engineering. J Integr Plant Biol 48:121–127

    Google Scholar 

  • Inomata S, Yokoyama M, Gozu Y, Shimizu T, Yanagi M (1993) Growth pattern and ginsenoside production of Agrobacterium-transformed Panax ginseng roots. Plant Cell Rep 12:681–686

    Article  CAS  Google Scholar 

  • Kumar V, Mehrotra N, Lal J, Gupta RC (2004) Pattern profiling of the herbal preparation picroliv using liquid chromatography-tandem mass spectrometry. J Chromatogr A 1045:145–152

    Article  PubMed  CAS  Google Scholar 

  • Kumar S, Singh J, Shah NC, Ranjan V (1997) Picrorhiza kurroa. In: Sharma A, Kumar A, Tewari R, Hasan S. A (eds) Indian medicinal and aromatic plants facing genetic erosion. Published by Director, CIMAP, Lucknow, pp 169–172

    Google Scholar 

  • Mano Y, Nabeshima S, Matsui C, Ohkawa H (1986) Production of tropane alkaloids by hairy root cultures of Scopolia japonica. Agric Biol Chem 50:2715–2722

    CAS  Google Scholar 

  • Mano Y, Ohkawa H, Yamada Y (1989) Production of tropane alkaloids by hairy root cultures of Duboisia leichhardtii transformed with Agrobacterium rhizogenes. Plant Sci 59:191–201

    Article  CAS  Google Scholar 

  • Morgan AJ, Cox PN, Turner DA, Peel E, Davey MR, Gartland KMA, Mullingan BJ (1987) Transformation of tomato using an Ri plasmid vector. Plant Sci 49:37–49

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nautiyal DP, Prakash V, Chauhan RS, Purohit H, Nautiyal MC (2001) Assessment of germinability, productivity, and cost benefit analysis of Picrorhiza kurrooa cultivated at lower altitudes. Curr Sci 81:579–585

    Google Scholar 

  • Porter JR (1991) Host range and implications of plant infection by Agrobacterium rhizogenes. Crit Rev Plant Sci 10:387–421

    Article  Google Scholar 

  • Rahman LU, Ikenaga T, Kitamura Y (2004) Penicillin derivatives induce chemical structure-dependent root development, and application for plant transformation. Plant Cell Rep 22:668–677

    Article  Google Scholar 

  • Ram VJ (2001) Herbal preparations as a source of hepatoprotective agents. Drug News Perspect 14:353–363

    Article  PubMed  CAS  Google Scholar 

  • Thyagarajan S, Jayaram S, Gopalakrishnan V, Hari R, Jeyakumar P, Sripathi M (2002) Herbal medicines for liver diseases in India. J Gastroenterol Hepatol 17 Suppl (3):S370–S376

    Article  Google Scholar 

  • Toivonen L, Ojala M, Kauppinen V (1991) Studies on the optimization of the growth and indole alkaloid production by hairy root cultures of Catharanthus roseus. Biotechnol Bioeng 37:673–680

    Article  CAS  PubMed  Google Scholar 

  • Torregrosa L, Iocco P, Thomas MR (2002) Influence of Agrobacterium strain, culture medium and cultivar on the transformation efficiency of Vitis vinifera L. Am J Enol Vitic 53:183–190

    CAS  Google Scholar 

  • Verma PC, Singh D, Rahman LU, Gupta MM, Banerjee S (2002) In vitro studies in Plumbago zeylanica: rapid micro propagation and establishment of higher plumbagin yielding hairy root cultures. J Plant Physiol 159:547–552

    Article  CAS  Google Scholar 

  • Verma PC (2003) Biotechnological approaches for in vitro production of picroliv from Agrobacterium rhizogenes mediated hairy root cultures of Picrorhiza kurroa Royle ex Benth. PhD Thesis, Lucknow University, Lucknow

  • Wysokinska H, Chmiel A (1997) Transformed root cultures for biotechnology. Acta Biotechnol 17:131–159

    Article  CAS  Google Scholar 

  • Yoshimatsu K, Satake M Shimomura K (1990) Determination of cardinolides in hairy root cultures of Digitalis lantana by enzyme linked immunoassay. J Nat Prod 53:1498–1502

    Article  PubMed  CAS  Google Scholar 

  • Zehra M, Banerjee S, Sharma S, Kumar S (1999) Influence of Agrobacterium rhizogenes strains on biomass and alkaloid productivity in hairy root lines of Hyoscyamus Muticus and H. albus. Planta Med 65:60–63

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The authors wish to express their sincere thanks to Dr. Sushil Kumar (Ex-Director, CIMAP, Lucknow) for providing the necessary research facilities and constant encouragement. Heartfelt thanks are also due to Dr. R Pal (Senior Scientist, CDRI, Lucknow) for his constructive suggestions and requisite guidance.

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Authors and Affiliations

  1. National Botanical Research Institute, Lucknow, UP, 226001, India

    Praveen Chandra Verma

  2. Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, UP, 26015, India

    Laiq ur Rahman, Arvind Singh Negi, Dharm Chand Jain & S. P. S. Khanuja

  3. Plant Tissue Culture Division, Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, UP, 26015, India

    Suchitra Banerjee

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  1. Praveen Chandra Verma
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  2. Laiq ur Rahman
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  3. Arvind Singh Negi
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  4. Dharm Chand Jain
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  5. S. P. S. Khanuja
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  6. Suchitra Banerjee
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Correspondence to Suchitra Banerjee.

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CIMAP Publication No.: 2007-28J

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Verma, P.C., ur Rahman, L., Negi, A.S. et al. Agrobacterium rhizogenes-mediated transformation of Picrorhiza kurroa Royle ex Benth.: establishment and selection of superior hairy root clone. Plant Biotechnol Rep 1, 169–174 (2007). https://doi.org/10.1007/s11816-007-0029-0

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  • DOI: https://doi.org/10.1007/s11816-007-0029-0

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