Improved Agrobacterium rhizogenes-mediated hairy root culture system of Withania somnifera (L.) Dunal using sonication and heat treatment.

Chandrasekaran Thilip,Chellappan Soundar Raju,Kandhan Varutharaju,Abubakker Aslam,Appakan Shajahan

doi:10.1007/s13205-015-0297-2

Abstract

An improved Agrobacterium rhizogenes-mediated genetic transformation of Withania somnifera (L.) Dunal was developed using the bacterial strain R1000 with leaf segment explants of in vitro raised plantlets. Out of the three strains used (R1000, MTCC 2364 and MTCC 532), the strain R1000 proved to be more efficient than others. Among the different conditions tested, the highest (93.3 %) transformation rate was observed after 3 weeks when the explants were subjected to sonication (15 s) and heat treatment (41 °C for 5 min). Transgenic status of the hairy roots was confirmed by PCR using rol B-specific primers. HPLC analysis showed the ability of hairy roots to synthesize withaferin A and withanolide A, both steroidal lactones of medicinal value. This protocol offers new avenue in A. rhizogenes-mediated hairy root induction and is useful for large-scale production of these bioactive compounds from W. somnifera.

Highlights

Withania somnifera (L.) Dunal (Solanaceae) is an important medicinal plant which yields pharmaceutically active compounds called withanolides (Praveen and Murthy2012)
Effect of A. rhizogenes strains on hairy root induction
Influence of A. rhizogenes strain on hairy root induction frequency has been documented earlier in several plant species (Sujatha et al 2013)

Summary

Introduction

Withania somnifera (L.) Dunal (Solanaceae) is an important medicinal plant which yields pharmaceutically active compounds called withanolides (Praveen and Murthy2012). Withania somnifera (L.) Dunal (Solanaceae) is an important medicinal plant which yields pharmaceutically active compounds called withanolides Roots of W. somnifera contain major withanolides like withaferin A and withanolide A and they possess specific therapeutic properties. Withaferin A is recognized for its anticancer properties and is reported to inhibit cell growth of various human cancer cell lines including lung cancer—NCl-H46 (Choudhary et al 2013). Withanolide A has recently been credited with a neotropic agent for recovery from nervous degeneration dendrites formation and its branching, and it can be a promising compound for the treatment of neural degeneration types of disease like Parkinson’s and Alzheimer’s disease (Kuboyama et al 2005; Sabir et al 2011). Other biological activities of W. somnifera include cholinesterase inhibition (Choudhary et al 2004, 2005), anti-inflammatory via COX-2 enzyme inhibition (Jayaprakasam and Nair 2003), antibacterial activity and sex hormone deficiency regulation (Kiasalari et al 2009), antiglycation (Maurya et al 2008), and antipyretic activities (Ali et al 1997)

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Results

Conclusion