Journal
3 BIOTECH
Volume 5, Issue 6, Pages 949-956Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s13205-015-0297-2
Keywords
Agrobacterium rhizogenes; Withania somnifera; Hairy roots; Withanolides; Sonication; Heat treatment
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Funding
- University Grants Commissions (UGC), Govt. of India, New Delhi [F. No. 42-946/2013-SR]
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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 degrees 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.
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