Improved Agrobacterium rhizogenes-mediated hairy root culture system of Withania somnifera (L.) Dunal using sonication and heat treatment.
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.
Plant Molecular Biology Laboratory, Department of Botany, Jamal Mohamed College, Tiruchirappalli, 620 020, Tamil Nadu, India.
Chandrasekaran
C
Thilip
Plant Molecular Biology Laboratory, Department of Botany, Jamal Mohamed College, Tiruchirappalli, 620 020, Tamil Nadu, India.
Chellappan
C
Soundar Raju
Plant Molecular Biology Laboratory, Department of Botany, Jamal Mohamed College, Tiruchirappalli, 620 020, Tamil Nadu, India.
Kandhan
K
Varutharaju
Plant Molecular Biology Laboratory, Department of Botany, Jamal Mohamed College, Tiruchirappalli, 620 020, Tamil Nadu, India.
Abubakker
A
Aslam
Plant Molecular Biology Laboratory, Department of Botany, Jamal Mohamed College, Tiruchirappalli, 620 020, Tamil Nadu, India. shajahan.jmc@gmail.com.
Appakan
A
Shajahan
Agrobacterium rhizogenes; Hairy roots; Heat treatment; Sonication; Withania somnifera; Withanolides