Vegetos
(An International Journal of Plant Research & Biotechnology)
(eISSN: 2229-4473)
Enhanced production of withanolide A in Agrobacterium rhizogenes-induced hairy root cultures of Withania somnifera (L.) Dunal compared to in vitro roots
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Nadaf Jafar Fakirasab, Nishani Sandhyarani, Narayanapur Vijayakumar B., Naik N. Jayarama, Chandalinga
Research Article | Published: 07 August, 2026
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Keywords: Hairy root culture, Withanolide A, n Agrobacterium rhizogenesn , In vitro biotechnology, n Withania somniferan
Abstract
Withania somnifera (L.) Dunal is a medicinally important plant recognized for its pharmacologically valuable withanolides, particularly withanolide A. However, conventional cultivation and non-transformed in vitro root cultures often exhibit limited and inconsistent metabolite accumulation. The present study aimed to establish Agrobacterium rhizogenes-mediated hairy root cultures and comparatively evaluate withanolide A production in transformed hairy roots and non-transformed in vitro roots. Hairy roots were induced from in vitro-grown leaf explants using A. rhizogenes strain MTCC 532 under optimized transformation conditions. The transformed roots exhibited characteristic hormone-independent growth, profuse lateral branching and rapid proliferation. Molecular confirmation by PCR amplification of the rolC gene verified stable integration of Ri T-DNA into the transformed root genome. Quantitative analysis using high-performance liquid chromatography (HPLC) demonstrated significantly higher withanolide A accumulation in hairy root cultures (0.880 mg g− 1 DW) than in non-transformed in vitro roots (0.162 mg g− 1 DW), representing an approximately 5.4-fold increase. The enhanced metabolite accumulation is likely associated with Ri T-DNA-mediated transformation and the altered metabolic activity of hairy root cultures. The findings demonstrate that A. rhizogenes-induced hairy root cultures provide a stable and efficient platform for improving withanolide A production in W. somnifera. This study establishes an optimized transformation protocol and provides a comparative evaluation of transformed hairy roots and non-transformed in vitro roots, highlighting the potential of hairy root biotechnology for sustainable production of valuable withanolides and future metabolic engineering applications.
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Author Information
Department of Plantation, Spices, Medicinal and Aromatic Crops (PSMA), College of Horticulture, Bagalkot, University of Horticultural Sciences, Bagalkot, India