Abstract
Plectranthus amboinicus (Lour.) Spreng is a perennial plant from the mint family with aromatic, succulent leaves and several health benefits. Multiple shoot regeneration was accomplished in vitro using nodal segments (NS) explants of P. amboinicus pretreated with 0, 0.5, 5, 25, 50, and 100 mu M thidiazuron (TDZ) for 4 h, then transferred to a growth regulator-free media. After 8 weeks of growth, NS explants pre-treated with 25 mu M TDZ for 4 h and then transferred to TDZ-free Murashige and Skoog (MS) media produced the greatest number of shoots (27.3 per NS) with the longest average shoot length (4.9 mm) in 97.2% of cultures. On the same medium, regeneration of roots in most of the P. amboinicus shoots occurred spontaneously. The in vitro-regenerated P. amboinicus plantlets were adequately hardened off and adapted to the ex-vitro environment with a 90% survival rate. Total phenolic, tannin, and flavonoid contents, as well as 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging/antioxidant activity, were significantly higher in in vitro-regenerated plants than in ex vitro-plants. Flow cytometry (FCM) analysis validated the nuclear genome stability of the in vitro generated plants, which assessed their nuclear DNA content and found it to be comparable in genome size to that of the field-grown plants. The study found a quick and efficient method for in vitro multiplication of P. amboinicus which can aid to increased availability and accessibility of this plant species for various purposes. The genetic and phytochemical analysis of the in vitro propagated plants can also provide valuable insight into the plant's properties and potential applications, which can further assist in its preservation and sustainable usage.