Bhaswatimayee Mahakur, Aryaputri Bedaprabha, Sandeep Kumar Mohanta, Sanjay Kumar Madkami, Debasis Mitra, Arpita Moharana, Swasti Swagatika Swain, Durga Prasad Barik

An improved protocol for inducing hairy roots in Vitex negundo using Agrobacterium rhizogenes

Abstract Vitex negundo L. contains numerous secondary metabolites in its roots. This experiment aimed to induce the growth of hairy roots that accumulate large quantities of secondary metabolites. The Agrobacterium rhizogenes (MTCC 532) strain was used to induce hairy roots in Vitex negundo explants. The transformation frequency was increased by adjusting the infection time, as well as the concentrations of the bacterial suspension and acetosyringone, and the co-cultivation period. In vitro leaf explants were collected from two-month-old mature shoot cultures, which exhibited the highest transformation frequency of 63.33%. Of the five infection durations and three acetosyringone concentrations tested, the highest transformation frequency was achieved with a 45-minute infection and 100 µM acetosyringone, respectively. A two-day co-cultivation period was found to be appropriate, and the explants were inoculated in hormone-free Murashige and Skoog medium to promote the spontaneous development of hairy roots.

Doi https://doi.org/10.35513/Botlit.2025.3.1

Keywords acetosyringone, Agrobacterium rhizogenes, secondary metabolites, Vitex negundo

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