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Genetic stability and phytochemical profiling of the in vitro regenerated plants of Angelica glauca Edgew.: an endangered medicinal plant of Himalaya

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Abstract

The present study describes the first successful report on in vitro propagation through direct organogenesis for multiple shoot induction of Angelica glauca. Rhizomes were used as explant, and maximum shoot multiplication was observed on MS medium supplemented with 6-Benzylaminopurine 8.0 µM and Indole-3-acetic acid 0.1 µM. Roots were observed within 14 days in the MS medium enriched with 0.5 µM IAA and 0.1 µM Naphthalene acetic acid (NAA) with an average production of 4.2 roots per shoot. Rooted plantlets were successfully hardened under greenhouse conditions and subsequently established in field, with a recorded survival rate of 72% after 45 days. The total phenolic content showed significant difference (p < 0.05) between in vitro raised plants (5.87 mM AAE/ g DW) and control (2.36 mM AAE/ g DW). Antioxidant activity, calculated through two in vitro assays, i.e. 1,1-diphenyl-2 picrylhydrazyl (DPPH) radical scavenging and Ferric Reducing Antioxidant Power (FRAP) assays revealed higher antioxidant activity in in vitro grown plants in comparison to control plants. Essential oil constituent’s analysis was also carried out in control and in vitro raised plants. Thirty-one compounds were identified in the oil samples through Gas chromatography (GC) and gas chromatography–mass spectrometry (GC–MS) analysis also identified 31 compounds in the essential oil, representing 98.1–98.7% of total oil compositions. The major components of the essential oils were (Z)-ligustilide (51.1–51.5%), (Z)-butylidene phthalide (31.2–31.6%), (E)-butylidene phthalide (2.6–2.9%) and (E)-ligustilide (2.1–1.8%). Genetic stability of in vitro raised plants, evaluated using 20 Inter Simple Sequence Repeats primers, proved true to typeness of in vitro raised plants.

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Abbreviations

MS:

Murashige and Skoog medium

BAP:

6-Benzylaminopurine

TDZ:

Thidiazuron

IAA:

Indole-3-acetic acid

NAA:

Naphthaleneacetic acid

Amsl:

Above mean sea level

ISSR:

Inter simple sequence repeats

GC–MS:

Gas chromatography–mass spectrometry

DPPH:

1, 1-Diphenyl-2- picrylhydrazyl

FRAP:

Ferric reducing antioxidant power

References

  • Adams RP (2009) Identification of essential oil components by gas chromatography/mass spectrometry, 4th edn. Allured Business Media, Illinois

    Google Scholar 

  • Agrawal VS (1986) Economic plants of India. Kailash Prakasn, Calcutta, p 22

    Google Scholar 

  • Amiri S, Kazemitabar SK, Ranjbar GA, Azadbakht M (2011) In vitro propagation and whole plant regeneration from callus in Datura (Datura stramonium. L). Afr J Biotechnol 10(3):442–448

    CAS  Google Scholar 

  • Amoo SO, Aremu AO, Van Staden J (2012) In vitro plant regeneration, secondary metabolite production and antioxidant activity of micropropagated Aloe arborescens mill. Plant Cell Tissue Org Cult 111:345–358

    Article  CAS  Google Scholar 

  • Anonymous (1985). The wealth of India—raw materials, vol 1. Publication and Information Directorate (CSIR), New Delhi, pp 275–276

    Google Scholar 

  • Anonymous (2003) CAMP report: conservation assessment and management prioritization for the medicinal plants of Jammu & Kashmir. Himachal Pradesh & Uttaranchal. FRLHT, Bangalore

    Google Scholar 

  • Bairu MW, Aremu AO, Van Staden J (2011) Somaclonal variation in plants: causes and detection methods. Plant Growth Regul 63:147–173

    Article  CAS  Google Scholar 

  • Behera SK, Rajasekaran C, Payas S, Fulzele DP, Doss CGP, Siva R (2017) In vitro flowering in Oldenlandia umbellata. J Ayurveda Integr Med. https://doi.org/10.1016/j.jaim.2017.02.011

    Article  PubMed  PubMed Central  Google Scholar 

  • Benzie IFF, Strain JJ (1996) The ferric reducing ability of plasma (FRAP) as a measure of Antioxident power the FRAP assay. Anal Biochem 239:70–76

    Article  CAS  PubMed  Google Scholar 

  • Bhattacharyya P, Kumaria S, Job N, Tandon P (2015) Phytomolecular profiling and assessment of antioxidant activity within micropropagated plants of Dendrobium thyrsiflorum: a threatened, medicinal orchid. Plant Cell Tissue Org Cult 122:535–550

    Article  CAS  Google Scholar 

  • Bisht AK, Manjkhola S, Joshi M (2003) Comparative Account of Two High Value Species of Himalayas: Angelica glauca Edgew. and Angelica archangelica. Linn Indian For 129(10):1241–1248

    Google Scholar 

  • Bisht AK, Bhatt A, Dhar U (2015) Note on somatic embryogenesis and synthetic seed production in Angelica glauca: a valuable medicinal plant of Himalaya. J Med Plant Res 9(12):419–425

    Article  Google Scholar 

  • Braca A, Tommasi ND, Bari LD, Pizza C, Politi M, Morelli I (2001) Antioxidant principles from Bauhinia terapotensis. J Nat Prod 64:892–895

    Article  CAS  PubMed  Google Scholar 

  • Butola JS, Badola HK (2004) Effect of pre-sowing treatment on seed germination and seedling vigour in Angelica glauca, a threatened medicinal herb. Curr Sci 87:796–799

    Google Scholar 

  • Butola JS, Vashistha RK (2013) An overview on conservation and utilization of Angelica glauca Edgew. in three Himalayan States of India. Med Plants 5:171–178

    Google Scholar 

  • Butola JS, Vashistha RK, Samant SS, Malik AR (2010) Technology for propagation and cultivation of Angelica glauca Edgew.: a threatened high value Himalayan medicinal cum edible herb. Med Plant 2(1):67–72

    Google Scholar 

  • Chavan JJ, Gaikwad NB, Umdale SD (2014) Efficiency of direct and indirect shoot organogenesis, molecular profiling, secondary metabolite production and antioxidant activity of micropropagated Ceropegia santapaui. Plant Growth Regul 72:1–15

    Article  CAS  Google Scholar 

  • Chung JW, Choi RJ, Seo EK, Nam JW, Dong MS, Shin EM, Guo LY, Kim YS (2012) Anti-inflammatory effects of (Z)-ligustilide through suppression of mitogen-activated protein kinases and nuclear factor-k B activation pathways. Arch Pharm Res 35:723–732

    Article  CAS  PubMed  Google Scholar 

  • Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15

    Google Scholar 

  • Feng Z, Lu Y, Wu X, Zhao P, Li J, Peng B, Qian Z, Zhu L (2012) Ligustilide alleviates brain damage and improves cognitive function in rats of chronic cerebral hypoperfusion. J Ethnopharmacol 144:313–321

    Article  CAS  PubMed  Google Scholar 

  • Giri L, Dhyani P, Rawata S, Bhatta ID, Nandi SK, Rawal RS, Pande V (2012) In vitro production of phenolic compounds and antioxidant activity in callus suspension cultures of Habenaria edgeworthii: A rare Himalayan medicinal orchid. Ind Crop Prod 39:1–6

    Article  CAS  Google Scholar 

  • Kirtikar KR, Basu BD (1988) Indian medicinal plants, Allahabad. Indian Med Plants 2:1214–1215

    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 

  • Nadeem M, Kumar A, Nandi SK, Palni LMS (2001) Tissue culture of medicinal plants with particular ref erence to Kumaun Himalaya. In: Proceedings of the workshop on Himalayan medicnal plants-potentialand prospects, Kosi-Katarmal, Almora, 5–7 Nov 1998

  • Nei M, Li WH (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Nat Acad Sci USA 76:5269–5273

    Article  CAS  PubMed  Google Scholar 

  • Pandey M, Dhar U, Samant SS, Thengane SR (2011) Recurrent somatic embryogenesis and plant regeneration in Angelica glauca Edgew., a critically endangered medicinal plant of the Western Himalaya. J Hortic Sci Biotechnol 86(5):493–498

    Article  Google Scholar 

  • Purohit VK, Andola HC, Haider SZ, Tiwari D, Bahuguna YM, Gairola KC, Arunachalam K (2015) Essential oil constituents of Angelica glauca Edgew. roots: an endangered species from Uttarakhand Himalaya (India). Natl Acad Sci Lett. https://doi.org/10.1007/s40009-015-0395-z

    Article  Google Scholar 

  • Rawat JM, Rawat B, Agnihotri RK, Chandra A, Nautiyal S (2013a) In vitro propagation, genetic and secondary metabolite analysis of Aconitum violaceum Jacq.—a threatened medicinal herb. Acta Physiol Plant. https://doi.org/10.1007/s11738-013-1294-x

    Article  Google Scholar 

  • Rawat JM, Rawat B, Chandra A, Nautiyal S (2013b) Influence of plant growth regulators on indirect shoot organogenesis and secondary metabolite production in Aconitum violaceum Jacq. Afr J Biotechnol 12(44):6287–6293

    Article  CAS  Google Scholar 

  • Samantaray S, Maiti S (2008) Rapid plant regeneration and assessment of genetic fidelity of in vitro raised plants in Aloe barbadensis Mill: using RAPD markers. Acta Bot Gallica 155(3):427–434

    Article  CAS  Google Scholar 

  • Sharma RN, Deshpande SG, Joseph M (1990) Evaluation of natural essential oil of Angelica glauca Edgew. of different geographical regions. Indian Perfum 34:196–198

    Google Scholar 

  • Sharma E, Gaur K, Punetha H, Gaur AK (2012) In vitro regeneration of Aconitum balforii stapf: a rare medicinal herb from Himalayan alpine through root explant. Res J Med Plant 6:318–325

    Article  CAS  Google Scholar 

  • Shinde AN, Malpathak N, Fulzele DP (2010) Determination of isoflavone and antioxidant activity in Psoralea corylifolia L. callus cultures. Food Chem 118:128–132

    Article  CAS  Google Scholar 

  • Siva R, Rajasekaran C, Mudgal G (2009) Induction of somatic embryogenesis and organogenesis in Oldenlandia umbellata L., a dye-yielding medicinal plant. Plant Cell Tissue Org Cult 98:205–211

    Article  CAS  Google Scholar 

  • Siva R, Mayes S, Behera SK, Rajasekaran C (2012) Anthraquinones dye production using root cultures of Oldenlandia umbellata L. Ind Crop Prod 37:415–419

    Article  CAS  Google Scholar 

  • Swain T, Hillis WE (1959) The phenolic constituents of Prunus domestica L. - the quantitative analysis of phenolic constituents. J Sci Food Agric 10:63–68

    Article  CAS  Google Scholar 

  • Thangavel P, John Britto S, Senthilkumar SR (2011) Adventitious shoot regeneration from leaf explants of the valuable medicinal herb Plectranthus barbatus Andrews. Afr J Biotechnol 10(43):8562–8569

    CAS  Google Scholar 

  • Vashistha R, Nautiyal BP, Nautiyal MC (2006) Conservation status and morphological variations between populations of Angelica glauca Edgew. and Angelica archangelica Linn. in Garhwal Himalaya. Curr Sci 91(11):1537–1542

    Google Scholar 

  • Vashistha RK, Butola JS, Nautiyal BP, Nautiyal MC (2010) Phenological attributes of Angelica glauca and A. archangelica expressed at two different climatic zones in Western Himalaya. Open Access J Med Aromat Plants 1:7–12

    Google Scholar 

  • Verma SK, Yucesan B, Sahin G, Gurel E (2014) Embryogenesis, plant regeneration and cardiac glycoside determination in Digitalis ferruginea subsp ferruginea L. Plant Cell Tissue Org Cult 119:625–634

    Article  CAS  Google Scholar 

  • Wong C, Li H, Cheng K, Chen F (2006) A systematic survey of antioxidant activity of 30 Chinese medicinal plants using the ferric reducing antioxidant power assay. Food Chem 97:705–711

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Authors are thankful to the Director of the Institute for providing necessary facilities to carry out this work. The financial support to the author Janhvi M. Rawat from SERB, DST, Govt. of India (SB/YS/LS-69/2014), is gratefully acknowledged.

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Author notes

  1. Ashok Kumar Dhakad

    Present address: Department of Forestry and Natural Resources, College of Agriculture, Punjab Agricultural University, Ludhiana, Punjab, India

Authors and Affiliations

  1. Systematic Botany Discipline, Botany Division, Forest Research Institute, Dehradun, Uttarakhand, India

    Janhvi Mishra Rawat, Ashok Kumar Dhakad & Anup Chandra

  2. Genetics and Tree Propagation Division, Forest Research Institute, Dehradun, Uttarakhand, India

    Aakriti Bhandari & Ajay Thakur

  3. Society for the Conservation of Nature, Parnkuti, Anantpur, University Road, Rewa, MP, India

    Susmita Mishra

  4. Department of Agriculture and Forestry, Graphic Era Hill University, Dehradun, India

    Balwant Rawat

Authors
  1. Janhvi Mishra Rawat
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  2. Aakriti Bhandari
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  3. Susmita Mishra
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  4. Balwant Rawat
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  5. Ashok Kumar Dhakad
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  7. Anup Chandra
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Contributions

Conceived and designed the experiments: JMR, AB, AT. Performed the experiments: JMR, AB, SM, AKD. Data recorded: JMR, AB, SM. Analyzed the data: JMR, AB, SM,AKD. Contributed materials/analysis tools: JMR, AKD. Wrote the paper: JMR, BR. Design of figures: JMR, BR, SM. Supervision of work: AT, AC. Plagiarism check was done AKD.

Corresponding author

Correspondence to Balwant Rawat.

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The authors have declared that no competing interests exist.

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Communicated by Sergio J. Ochatt.

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Rawat, J.M., Bhandari, A., Mishra, S. et al. Genetic stability and phytochemical profiling of the in vitro regenerated plants of Angelica glauca Edgew.: an endangered medicinal plant of Himalaya. Plant Cell Tiss Organ Cult 135, 111–118 (2018). https://doi.org/10.1007/s11240-018-1448-z

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