An efficient method of in-vitro micropreparation of piper longum from lateral bud explant, said method comprising the steps of:-i. cutting the lateral buds from the stem segments under laminar flow hood and dipping them in hydrogen chloride (0.002 - 0.2%) for 3 minutes and rinsing it with autoclaved distilled water;ii. subjecting the lateral buds to surface sterilisation by dipping in sodium hypochloride (NaOCl) (0.5 - 3%) for 4 minutes and again giving three thorough rinses with autoclave distilled water;iii. inoculating the surface sterilised lateral buds in a culture initiation medium supplemented with growth hormones;iv. incubating the inoculated explants in dark at 23°C + I °C, for 7-15 days to ntoduce. 6-7 root primordia from the basal cut end of all the lateral buds;v. transferring the incubated cultures of step iv, in light intensity of 2000-4000 lux and providing a photo period of 16 hours followed by a dark period of 8 hours daily ;vi. allowing each root to grow to an average size of 1.5 cm in 45 days; through direct rhizogenesis (organogenesis);vii. cutting each root of step vi, into 0.5 cm long segments aseptically under LF;viii. culturing the 18 segments obtained as in step vii onto a shoot induction medium supplemented with the growth hormone ;ix. keeping the cultures of step viii, at 23° C under light intensity of 2000-4000-lux for a photo period of 16 hours followed by a dark period of 8 hours to produce about 50 - 55 adventitious shoot buds; x. cutting each culture clump into two pieces after 45 days of interval and sub culturing each piece in a separate culture bottle containing 25 ml of the same medium, and growing them under similar physical conditions as mentioned step ix, for 45 days for shoot elongation;xi. separating 2.5 - 3 cm long shoots with 2-3 nodes from the culture clumps using a sterile scalpel under LF;xii. culturing 10 separated shoots of step xi, in a wide mouth bottle containing rooting medium supplemented with growth hormone;xiii. rooting the regenerated shoots of step xii, under reduced light intensity of 1000 - 3000 lux for 28 days, with 16 hour photoperiod and temperature 23° C ± 1° C ;xiv. removing the micropropagated plants carefully from the agar contained in the sub-culturing medium;xv. washing the roots of the micropropagated plants of step xiv, under tap water to remove any trace of agar adhered to the roots and subjecting to hardening by known methods.

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