*Article* **In Vitro Propagation of** *Aconitum violaceum* **Jacq. ex Stapf through Seed Culture and Somatic Embryogenesis**

**Abdul Hadi 1,2, Seema Singh 1, Shah Rafiq 1,\*, Irshad A. Nawchoo 2, Nasir Aziz Wagay 3, Eman A. Mahmoud 4, Diaa O. El-Ansary 5, Hanoor Sharma 6, Ryan Casini 7, Kowiyou Yessoufou <sup>8</sup> and Hosam O. Elansary 9,\***


**Abstract:** *Aconitum violaceum* Jacq. ex Stapf is a threatened medicinal plant with restricted global distribution. The highest frequency of seed germination was recorded on Murashige and Skoog's (MS) basal medium, supplemented with 0.5 mg L−<sup>1</sup> kinetin with a germination rate of 77.32% and mean germination time of 27 days. Among the various plant growth regulators examined, 0.1 mg L−<sup>1</sup> kinetin (Kn) + 0.5 mg L−<sup>1</sup> indole-3-acetic acid (IAA) proved to be effective for maximum embryogenic callus production (51.0%) within 31 days of inoculation. The conversion rate of somatic embryos into complete plantlets was highest in the MS medium augmented with 0.1 mg L−<sup>1</sup> Kn + 0.5 mg L−<sup>1</sup> IAA (68.00%), with an average root initiation time of 25 days. The rooted plantlets were subsequently hardened into jiffy pots with a combination of loamy soil, coco-peat, and vermicompost (1:1:1 *v*/*v*), and then transplanted into a greenhouse with a 60% survival rate. To our knowledge, this is the first study on direct in vitro propagation and embryogenic callus induction from seeds. The established regeneration protocol could be employed to propagate *A. violaceum* on a large scale in a short time. This would contribute significantly to its rapid propagation and germplasm conservation, and establish a framework for the domestication of this highly valued threatened medicinal plant.

**Keywords:** acclimatization; callusing; multiple shooting; seed germination; micropropagation
