Pueraria phaseoloides (Roxb.) Benth. Commonly known as tropical kudzu, is an ethno-medicinally significant climbing perennial legume of the Fabaceae family. Endemic to regions including the Chittagong Hill Tracts (CHT) of Bangladesh, it is heavily exploited for its bioactive iso-flavonoids, notably puerarin. The species currently faces localized threats due to overharvesting and low natural regeneration rates. To mitigate these conservation challenges and establish a sustainable supply for the pharmaceutical industry, an efficient in vitro seed germination and direct micropropagation protocol was established. Seed germination was optimized on Murashige and Skoog (MS) medium, achieving a maximum germination rate of 100% within 30-33 days when supplemented with 1.0 mg/l 6-benzylaminopurine (BAP) and 0.5 mg/l α-naphthaleneacetic acid (NAA). For multiple shoot buds (MSBs) induction via direct organogenesis, nodal segments exhibited the highest morphogenic response (80%) on MS medium fortified with 2.0 mg/l BAP and 2.0 mg/l indole-3-acetic acid (IAA), producing a maximum of 2.29 ± 0.41 MSBs per explant. Optimal shoot elongation (an increase of 3.07 ± 0.18 cm after 30d of culture) was recorded on MS medium containing 2.0 mg/l BAP and 1.0 mg/l NAA. For rhizogenesis, half strength MS medium supplemented with 2.0 mg/l indole-3-butyric acid (IBA) proved superior, inducing a 90% rooting frequency with an average of 6.17 ± 0.19 number of roots per shoot and a maximum root length of 3.26 ± 0.22 cm. Fully regenerated seedlings were successfully acclimatized ex vitro with a 70% survival rate. This highly reproducible propagation protocol provides a robust platform for the rapid multiplication, conservation and commercial cultivation of P. phaseoloides.