Development of plant tissue culture system for Nyctanthes arbor-tristis L. in a controlled environmental system from shoot-tip culture and genetic homogeneity evaluation utilizing inter simple sequence repeat (ISSR) and start codon targeted (SCoT) markers

Abstract

An efficient in vitro shoot development profile is developed from the shoot-tip culture of N. arbor-tristis L. To evaluate cytokinin efficacy for manifold shoot regeneration from the shoot-tip, it was cultured on tissue culture Murashige and Skoog medium augmented with various strengths of cytokinin. The results emphasized that medium augmented with 5 mg l−1 BA was found most appropriate for the in vitro shoot development from the cultured explant. This media supported induction of the maximum shoot regeneration efficiency (75.33 ± 1.76), maximum number of shoots/explant (28.23 ± 0.52) and shoot length (4.53 ± 0.12 cm) without callus formation. The effect of variation in nitrogen content (KNO<inf>3</inf> and NH<inf>4</inf>NO<inf>3</inf>) in medium on shoot regeneration potential was studied. The optimum regeneration from selected explant was achieved on medium compositions M<inf>3</inf> (1900 mg l−1 KNO<inf>3</inf> + 1650.00 mg l−1 NH<inf>4</inf>NO<inf>3</inf>) and M<inf>9</inf> (1650.00 mg l−1 NH<inf>4</inf>NO<inf>3</inf> + 1900 mg l−1 KNO<inf>3</inf>) respectively. The best in vitro root induction (69.50 ± 0.76%), the maximum roots/shoot (10.31 ± 0.82), and its optimum length (2.96 ± 0.14 cm) were achieved once microshoots were sub-inoculated on full-strength Murashige and Skoog medium after being pulse treated with 300 mg l−1 IBA for up to 30 min. The molecular authentication of regenerated shoots to the mother plant was confirmed using the inter simple sequence repeat (ISSR) and start codon targeted (SCoT) primers. Thus, this protocol may aid the consistent production of quality planting stocks, which can be used for transformation study and in other ethnomedicinal purposes. © The Author(s), under exclusive licence to Springer Nature B.V. 2025.