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ISSR and RAPD based evaluation of genetic fidelity and active ingredient analysis of regenerated plants of Picrorhiza kurroa

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Abstract

Genetic stability and phytochemical analysis of in vitro established plants of Picrorhiza kurroa Royle ex Benth, have been carried out. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) markers were used to assess the genetic fidelity of tissue culture products including three adventitious shoots from three calli and 6 months old tissue culture raised plants growing in green house condition with mother plant. Apparent genetic variation was detected in the five types of plant materials. The percentage of polymorphic bands in the RAPD and ISSR analysis were 16.25 and 14.54 %, respectively. The genetic similarity was calculated on the basis of RAPD and ISSR data among the five types of plant materials and were ranged from 0.5 to 1.0 (mean 0.75) and 0.47 to 1.0 (mean 0.73), respectively. The similarity coefficient by both RAPD and ISSR analysis revealed that differences between tissue culture raised plants and mother plant was not remarkable, but notable differences were observed among three adventitious shoots regenerated from three calli. The phytochemical analysis of tissue culture raised products showed higher secondary metabolite (picrotin and picrotoxinin) content as compare to mother plant. The information gained on genetic stability/variability will be valuable for the large scale propagation and secondary metabolite production of P. kurroa.

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Abbreviations

MS:

Murashige and Skoog’s medium

BAP:

6-Benzylaminopurine

IAA:

Indole acetic acid

NAA:

α-Naphthaleneacetic acid

PCR:

Polymerase chain reaction

RAPD:

Random amplified polymorphic DNA

ISSR:

Inter simple sequence repeats

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Acknowledgments

Director of the Institute is thanked for providing facilities. The financial assistance to the author JMR from Department of Science and Technology (WOS/LS-191/2010), India is gratefully acknowledged. Sincere thanks goes to CSIR (09/560(0015)/2011-EMR-I) for financial and technical support.

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Authors and Affiliations

  1. Botany Division, Forest Research Institute, Dehradun, 248006, India

    Janhvi Mishra Rawat, Anup Chandra & Subhash Nautiyal

  2. GB Pant Institute of Himalayan Environment and Development, Kosi-Katarmal, Almora, 263 643, Uttarakhand, India

    Janhvi Mishra Rawat & Balwant Rawat

  3. Society for the Conservation of Nature, Anantapur University Road, Rewa, 486 002, Madhya Pradesh, India

    Balwant Rawat

  4. Plant Biotechnology Division, Central Institute of Medicinal and Aromatic Plants, Lucknow, 226 015, India

    Shakti Mehrotra

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  1. Janhvi Mishra Rawat
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  2. Balwant Rawat
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  3. Shakti Mehrotra
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  4. Anup Chandra
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  5. Subhash Nautiyal
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Correspondence to Balwant Rawat.

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Communicated by J. W. Sadowski.

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Rawat, J.M., Rawat, B., Mehrotra, S. et al. ISSR and RAPD based evaluation of genetic fidelity and active ingredient analysis of regenerated plants of Picrorhiza kurroa . Acta Physiol Plant 35, 1797–1805 (2013). https://doi.org/10.1007/s11738-013-1217-x

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