<p><i>Persicaria glabra</i> is an important medicinal plant with potent pharmacological properties. The in vitro propagation of <i>P. glabra</i> in Murashige and Skoog (MS) medium supplemented with 2.5 µM benzyl adenine (BA) was hindered by early leaf senescence and leaf yellowing symptoms, followed by drying of leaves with an increase in the number of subculture trials. The early leaf senescence may be attributed to the accumulation of the gaseous hormone ethylene in the culture vessels. To check the impact of ethylene in early leaf senescence, an ethylene-liberating compound, ethephon, was tested. For the regulation of ethylene accumulation, two ethylene inhibitors, silver thiosulfate (STS) and sodium nitroprusside (SNP), were examined at different concentrations (10, 20, 30, 40 µM) with 2.5 µM BA. Among the two ethylene inhibitors, STS was more effective, and the addition of 30 µM STS resulted in a reduced number of dried leaves (0.17 per culture) and the production of multiple shoots (8.28 shoots per culture). A higher amount of chlorophyll and lower hydrogen peroxide (H₂O₂) content were present in the STS-treated cultures. Healthy microshoots developed in 30 µM STS were selected for in vitro rooting experiments in different auxins, and 2.5 µM naphthaleneacetic acid (NAA) produced the highest number of roots and root length. The rooted plants were hardened and acclimatized in field conditions with 100% survival. The complete monomorphism by start codon targeted (SCoT) markers revealed genetic homogeneity between the regenerated plant and the mother plant.</p>