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Enhanced secondary metabolite production and pathway gene expression by leaf explants-induced direct root morphotypes are regulated by combination of growth regulators and culture conditions in Centella asiatica (L.) urban

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Abstract

Centella asiatica (L), a herbaceous plant belonging to the family Apiaceae, possesses great medicinal value owing to the presence of important and characteristic triterpenoids as secondary metabolites. These triterpenoid secondary metabolites are found in leaves in substantial quantities whereas negligible amounts may be detected sometimes in root tissues. In the resent study direct rhizogenesis was induced from C. asiatica leaf explants using different concentrations and combinations of auxins (IBA/IAA/NAA) leading to the production of distinct root morphotypes. A number of culture conditions such as pH, nature of carbon sources (glucose, fructose, mannitol and maltose) as well as concentrations of sucrose exhibited their strong influence in terms of induction of root morphotypes, accumulation of total secondary metabolites and expression of key pathway genes. Phytochemical profiling using HPLC revealed that all root morphotypes accumulated enhanced amounts of triterpenoids. The enhanced phytochemical accumulation was further validated by the coherent pattern of expression of key genes related to their biosynthetic pathway in root morphotypes. The results have revealed that the hormonal combinations in the culture media not only mediated differential morphogenic responses but also regulated secondary metabolites accumulation in non-transgenic rhizogenic roots. The results of the study are promising for the utilization of such in vitro root morphotypes. The root morphotypes may act as alternative bioresources for the production of industrially important and leaf associated asiaticosides and other important triterpenoids for the commercial purposes.

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References

  • Al-Hawamdeh FM, Shibli RA, Al-Qudah TS (2013) In vitro production of silymarin from Silybum marianum L. Med Aromat Plants 1:001. doi:10.4172/2167-0412.S1-00

    Google Scholar 

  • Aziz ZA, Davey MR, Power JB, Anthony P, Smith RM, Lowe KC (2007) Production of asiaticoside and madecassoside in Centella asiatica in vitro and in vivo. Biol Plant 51:34–42

    Article  CAS  Google Scholar 

  • Bahmani R, Karami O, Gholami M (2009) Influence of carbon sources and their concentrations on rooting and hyperhydricity of apple rootstock MM.106. World Appl Sci J 6:1513–1517

    CAS  Google Scholar 

  • Bose SK, Yadav RK, Mishra S, Sangwan RS, Singh AK, Mishra B, Srivastava AK, Sangwan NS (2013) Effect of gibberellic acid and calliterpenone on plant growth attributes, trichomes, essential oil biosynthesis and pathway gene expression in differential manner in Mentha arvensis L. Plant Physiol Biochem 66:150–158. doi:10.1016/j.plaphy.2013.02.011

    Article  CAS  PubMed  Google Scholar 

  • Chapman EJ, Estelle M (2009) Mechanism of auxin-regulated gene expression in plants. Annu Rev Genet 43:265–285

    Article  CAS  PubMed  Google Scholar 

  • Chaurasiya ND, Uniyal GC, Lal P, Misra L, Sangwan NS, Tuli R, Sangwan RS (2007) Analysis of with anolides in root and leaf of Withania somnifera by HPLC with photodiode array and evaporative light scattering detection. Phytochem Anal 19:148–154

    Article  Google Scholar 

  • Chaurasiya ND, Sangwan NS, Sabir F, Misra L, Sangwan RS (2012) Withanolide biosynthesis recruits both mevalonate and DOXP pathways of isoprenogenesis in ashwagandha Withania somnifera L. (Dunal). Plant Cell Rep 31:1889–1897

    Article  CAS  PubMed  Google Scholar 

  • Cui XH, Chakrabarty D, Lee EJ, Pae KY (2010) Production of adventitious roots and secondary metabolites by Hypericum perforatum L. in a bioreactor. Bioresour Technol 101:4708–4716

    Article  CAS  PubMed  Google Scholar 

  • Da Silva JAT (2004) The effect of carbon source on in vitro organogenesis of Chrysanthemum thin cell layers. Bragantia 631:65–177

  • Dacosta CT, Dealmeida MR, Ruedell CM, Schwambach J, Maraschin FS, Fett-Neto AG (2013) When stress and development go hand in hand: main hormonal controls of adventitious rooting in cuttings. Front Plant Sci 4:1–19

    Google Scholar 

  • Deduke C, Timsina B, Normore MDP (2012) Effect of environmental change on secondary metabolite production in lichen-forming fungi. International Perspectives on Global Environmental Change ISBN: 978-953-307-815-1

  • Gupta S, Shi X, Lindquist IE, Devitt N, Mudge J, Rashotte AM (2013) Transcriptome profiling of cytokinin and auxin regulation in tomato root. J Exp Bot 64:695–704

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Jeong GT, Woo JC, Park DH (2007) Effect of plant growth regulators on growth and biosynthesis of phenolic compounds in genetically transformed hairy roots of Panax ginseng C. A. Meyer. Biotechnol Bioprocess Eng 12:86–91

    Article  CAS  Google Scholar 

  • Kagmi H (1999) Effect of sugars on rooting of shoots of Japanese persimmon propagate. Plant Biotechnol 16:371–374

    Article  Google Scholar 

  • Karalija E, Parić A (2011) The effect of BA and IBA on the secondary metabolite production by shoot culture of Thymus vulgaris L. Biol Nyssana 2:29–35

    Google Scholar 

  • Kim OT, Kim MY, Hong MH, Ahn JC, Hwang B (2004) Stimulation of asiaticoside accumulation in the whole plant cultures of Centella asiatica (L.) Urban by elicitors. Plant Cell Rep 23:339–344

    Article  CAS  PubMed  Google Scholar 

  • Koperdakova J, Katkovcinova Z, Kosuth J, Giovannini A, Cellarova E (2009) Morphogenetic response to plant growth regulators in transformed and untransformed Hypericum perforatum L. clones. Acta Biol Crac Bot 51:61–70

    Google Scholar 

  • Lian ML, Chakrabarty D, Paek KY (2002) Effect of plant growth regulators and medium composition on cell growth and saponin production during cell-suspension culture of mountain ginseng (Panax ginseng C.A. Mayer). J Plant Biol 45:201–206

    Article  CAS  Google Scholar 

  • Luthra R, Sangwan RS, Sangwan NS (1993) Utilization of exogenously supplied primary precursors for oil synthesis in Cymbopogon species. Biol Plant 35:473–476

    Article  CAS  Google Scholar 

  • Mercy S, Nair RR, Ganesh D (2011) Influence of genotype on regeneration of adventitious roots from leaf derived callus of Centella asiatica. Biol Technol 2:50–56

    CAS  Google Scholar 

  • Mercy S, Sangeetha N, Ganesh D (2012) In vitro production of adventitious roots containing asiaticoside from leaf tissues of Centella asiatica (L.). In Vitro Cell Dev Biol Plant 48:200–207

    Article  Google Scholar 

  • Naik PM, Manohar SH, Praveen N, Murthy HN (2010) Effect of sucrose and pH levels on in vitro shoot regeneration from leaf explants of Bacopa monnieri and accumulation of bacoside A in regenerated shoots. Plant Cell Tissue Organ Cult 100:235–239

    Article  Google Scholar 

  • Narnoliya L, Rajakani R, Sangwan NS, Gupta V, Sangwan RS (2014) Comparative transcripts profiling of fruit mesocarp and endocarp relevant to secondary metabolism by suppression subtractive hybridization in Azadirachta indica (neem). Mol Biol Rep. doi:10.1007/s11033-014-3174-x

    PubMed  Google Scholar 

  • Pandey VP, Cherian E, Patani G (2010) Effect of growth regulators and culture conditions on direct root induction of Rauwolfia serpentine L. (Apocynaceae) benth by leaf explants. Trop J Pharm Res 9:27–34

    Article  CAS  Google Scholar 

  • Rafamantananaa MH, Rozetc E, Raoelisona GE, Cheuka K, Ratsimamangaa SU, Hubert P, Quetin-Leclercqb J (2009) An improved HPLC-UV method for the simultaneous quantification of triterpenic glycosides and aglycones in leaves of Centella asiatica (L.) Urb (APIACEAE). J Chromatogr B 877:2396–2402

    Article  Google Scholar 

  • Randriamampionona D, Diallo B, Rakotoniriana F, Rabemanantsoa C, Cheuk K, Corbisier AM, Mahillon J, Ratsimamanga S, Jaziri ME (2007) Comparative analysis of active constituents in Centella asiatica samples from Madagascar: application for ex situ conservation and clonal propagation. Fitoterapia 78:482–489

    Article  CAS  PubMed  Google Scholar 

  • Sabir F, Sangwan NS, Chaurasiya ND, Misra LN, Tuli R, Sangwan RS (2007) Micro-propagation of Withania somnifera L. accessions from axillary meristem for rapid propagation and withanolide production. J Herbs Spices Med Plant 13:123–133

    Article  CAS  Google Scholar 

  • Sabir F, Sangwan RS, Singh J, Pathak N, Mishra LN, Sangwan N S (2011) Biotransformation of withanolides by cell suspension cultures of Withania somnifera (Dunal)–Plant Biotechnology Reports-5, 12–134. doi:10.1007/s11816-011-0165-4

  • Sabir F, Mishra S, Sangwan RS, Jadaun JS, Sangwan NS (2013) Qualitative and quantitative variations in withanolides and expression of some pathway genes during different stages of morphogenesis in Withania somnifera Dunal. Protoplasma 250:539–549

    Article  CAS  PubMed  Google Scholar 

  • Sangwan RS, Chaurasiya ND, Lal P, Misra L, Uniyal GC, Tuli R, Sangwan NS (2007) Withanolide A biogeneration in in vitro shoot cultures of Ashwagandha (Withania somnifera Dunal), a main medicinal plant of ayurveda. Chem Pharm Bull 55:1371–1375

  • Sangwan RS, Chaurasiya ND, Lal P, Misra L, Tuli R, Sangwan NS (2008) Withanolide A is inherently de novo biosynthesized in roots of the medicinal plant Ashwagandha (Withania somnifera). Physiol Plant 133:278–287

    Article  CAS  PubMed  Google Scholar 

  • Sangwan RS, Tripathi S, Singh J, Narnoliya LK, Sangwan NS (2013) De novo sequencing and assembly of Centella asiatica leaf transcriptome for mapping of structural, functional and regulatory genes with special reference to secondary metabolism. Gene 525:58–76

    Article  CAS  PubMed  Google Scholar 

  • Seo PJ, Park CM (2009) Auxin homeostasis during lateral root development under drought condition. Plant Signal Behav 4:1002–1004

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Singh N, Luthra R (1988) Sucrose metabolism and essential oil accumulation during lemongrass (Cymbopogon flexuosus Stapf) leaf development. Plant Sci 57:127–133

    Article  CAS  Google Scholar 

  • Steinitz B (1999) Sugar alcohols display non osmotic roles in regulating morphogenesis and metabolism in plants that do not produce polyols as primary photosynthetic products. J Plant Physiol 155:1–8

    Article  CAS  Google Scholar 

  • Tominaga M, Sonobe S, Shimmen T (1998) Mechanism of inhibition of cytoplasmic streaming by auxin root hair cells of Hydrocharis. Plant Cell Physiol 39:1342–1349

    Article  CAS  Google Scholar 

  • Venverloo CJ (1976) The formation of adventitious organs III. A comparison of root and shoot formation on Nautilocalyx explants. J Plant Physiol 80:310–322

    CAS  Google Scholar 

  • Wang Y, Weathers PJ (2007) Sugars proportionately affect artemisinin production. Plant Cell Rep 26:1073–1081

    Article  PubMed  Google Scholar 

  • Yadav RK, Sangwan RS, Sabir F, Srivastava, AK and Sangwan NS (2014) Effect of prolonged water stress on specialized secondary metabolites, peltate glandular trichomes, and pathway gene expression in Artemisia annua L. Plant Physiology and Biochemistry 74C:70–83. doi:10.1016/j.plaphy.2013.10.023

  • Yang YK, Lee SY, Park WT, Park NI, Park SU (2010) Exogenous auxins and polyamines enhance growth and rosmarinic acid production in hairy root cultures of Nepeta cataria L. Plantomics 3:190–193

    CAS  Google Scholar 

  • Yoo NH, Kim OT, Kim JB, Kim SH, Kim YC, Bang KH, Hyun DY, Cha SW, Kim MY, Hwan B (2011) Enhancement of centelloside production from cultured plants of Centella asiatica by combination of thidiazuron and methyl jasmonate. Plant Biotechnol Rep 5:283–287

    Article  Google Scholar 

  • Zavattieri A, Lima M, Sobral V, Oliveira P, Costa A (2009) Effects of carbon source, carbon concentration and culture conditions on in vitro rooting of Pinus pinea L. microshoots. Acta Hort 812:173–180

    Google Scholar 

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Acknowledgments

JS and FS are thankful to CSIR, New Delhi and UGC, New Delhi for financial assistance in the form of senior research fellowship. The authors wish to express their sincere thanks to the director, CSIR-CIMAP, for constant encouragement and providing necessary facilities. The financial Grant from DBT, New Delhi to carry out above studies is gratefully acknowledged.

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Authors and Affiliations

  1. CSIR-Central Institute for Medicinal and Aromatic Plants (CSIR-CIMAP), Lucknow, 226015, UP, India

    Jyoti Singh, Farzana Sabir, Rajender Singh Sangwan, Lokesh K. Narnoliya & Neelam S. Sangwan

  2. Department of Biotechnology, Babasaheb Bhimrao Ambedkar University (BBAU), Central University, Rai Bareilly Road, Lucknow, 226025, UP, India

    Sangeeta Saxena

Authors
  1. Jyoti Singh
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  2. Farzana Sabir
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  3. Rajender Singh Sangwan
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  4. Lokesh K. Narnoliya
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  5. Sangeeta Saxena
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  6. Neelam S. Sangwan
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Corresponding author

Correspondence to Neelam S. Sangwan.

Additional information

Jyoti Singh and Farzana Sabir have equally contributed to this work.

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10725_2014_9931_MOESM1_ESM.ppt

Supplementary material HPLC profiles of (a-b) with respect to the four major bioactive authentic compounds resolved under standard conditions; (c), representative HPLC chromatogram of extract from the C. asiatica roots produced by induction of rhizogenesis by media supplementation with auxins (1mgl−1) IBA (PPT 277 kb)

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Singh, J., Sabir, F., Sangwan, R.S. et al. Enhanced secondary metabolite production and pathway gene expression by leaf explants-induced direct root morphotypes are regulated by combination of growth regulators and culture conditions in Centella asiatica (L.) urban. Plant Growth Regul 75, 55–66 (2015). https://doi.org/10.1007/s10725-014-9931-y

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  • DOI: https://doi.org/10.1007/s10725-014-9931-y

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