Optimizing Phytohormone Combinations in Murashige and Skoog Media for Enhanced Micropropagation of Ajuga bracteosa

Identification and conservation of endangered plant species are essential for safeguarding the flora of the region. Ajuga bracteosa (A. bracteosa), found in Pakistan's North West, holds immense medicinal potential due to its diverse active ingredients. However, unskilled overharvesting of this plant by uprooting entire herbs has drastically reduced its propagules, worsening its endangered status. The current study was conducted to optimize phytohormone levels in Murashige and Skoog media for micropropagation of A. bracteosa. In vitro micropropagation is an effective conservation technique for A. bracteosa due to its ability to preserve genetic diversity, enable rapid multiplication, produce disease-free plants, and facilitate long-term storage and utility in medicine. The study was divided into two experiments. In the first experiment, shoot organogenesis was studied in callus explants in the media containing 6-Benzylaminopurine (BAP) alone and combined with Auxins or Gibberellic Acid (GA₃). In the second experiment, these shoots were studied for root organogenesis in the media containing auxins Indole-3-acetic acid (IAA), Indole-3- butyric acid (IBA) and Naphthaleneacetic acid (NAA) at two levels (0.5 and 1.0 mg L^-1). In the first experiment, the medium containing BAP+NAA (3+1 mg L^-1) induced early and 100% shoots and 13.8 shoot explants^-1 with 4.1 cm length. In the second experiment, NAA (0.5 mg L^-1) induced the roots early and 100% with 7.2 roots shoot^-1 and 3.6 cm length. The results of the present study suggest that, for better regeneration of shoots, BAP+NAA (3+1 mg L^-1) should be used in the medium, while NAA (0.5 mg L^-1) is required to be added to the medium for better root regeneration.