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Applied Biochemistry and Biotechnology

, Volume 162, Issue 7, pp 1929–1937 | Cite as

Rapid In Vitro Production of Cloned Plants of Uraria picta (Jacq.) DC—A Rare Medicinal Herb in Long-Term Culture

  • Santosh Kumar Rai
  • Meena Sharma
  • Madhu Jain
  • Abhishek Awasthi
  • Dharmendra Kumar Purshottam
  • Narayanan Kuttanpillai Nair
  • Ashok Kumar SharmaEmail author
Article

Abstract

An efficient in vitro process for rapid production of cloned plants of Uraria picta has been developed employing nodal stem segments taken from field-grown plants. Explants showed bud-break followed by regeneration of shoots with restricted growth within 12 days on modified Murashige and Skoog’s medium supplemented with 0.25 mg l–1 each of 6-benzylaminopurine and indole-3-acetic acid and 25 mg l-1 adenine sulfate. Normal growth of shoots with good proliferation rate was achieved by reducing the concentrations of 6-benzylaminopurine and indole-3-acetic acid to 0.1 mg l-1 each and incorporating 0.5 mg l-1 gibberellic acid in the medium in which, on an average, 19.6 shoots per explant were produced. Further, during successive subcultures, increased concentrations of adenine sulfate (50 mg l-l) and gibberellic acid (2 mg l-l) along with the addition of 20 mg l-ldl-tryptophan were found conducive to control the problem of necrosis of shoots. In this treatment, several “crops” of shoots were obtained from single culture by repeated subculturing of basal portion of stalk in long-term. Isolated shoots rooted 100% in 0.25 mg l-1 indole-3-butyric acid. In vitro-raised plants after hardening in inorganic salt solution grew normally in soil and came to flowering. Genetic fidelity of in vitro-raised plants was ascertained by rapid amplified polymorphic DNA (RAPD) markers. Also, quantitative estimation of two isoflavonones in their root extracts further confirmed true-to-type nature of plantlets.

Keywords

Conservation Genetic fidelity In vitro cloning Nodal stem segments Rare medicinal herb 

Notes

Acknowledgments

The authors express gratitude to The Director, National Botanical Research Institute, Lucknow, for the facilities provided and Botanic Garden Conservation International, UK, for partial financial support. Thanks are also due to Dr. R.L.S. Sikarwar, Senior Research Officer, Deendayal Research Institute, Chitrakoot, for providing the plant material and Shri R.S. Tripathi and Shri O.P. Sharma for technical support.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Santosh Kumar Rai
    • 1
  • Meena Sharma
    • 1
  • Madhu Jain
    • 1
  • Abhishek Awasthi
    • 1
  • Dharmendra Kumar Purshottam
    • 1
  • Narayanan Kuttanpillai Nair
    • 1
    • 2
  • Ashok Kumar Sharma
    • 1
    Email author
  1. 1.Tissue Culture LaboratoryNational Botanical Research InstituteLucknowIndia
  2. 2.Conservation Biology and Molecular Taxonomy DivisionNational Botanical Research InstituteLucknowIndia

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