A procedure for in vitro multiplication of Saussurea lappa (Asteraceae) is described. On Murashige and Skoog's medium (MS) containing benzylaminopurine and gibberellin 3.5-fold shoot multiplication occurred every three weeks. Shoots rooted on MS containing 0.5 ~M naphthaleneacetic acid with 90% efficiency. The shoot cultures stored at 5°C in the dark for 12 months without an intervening subculture survived with 100% viability. The shoots cold stored for 6 months or more showed higher rates of multiplication under culture room conditions than the untreated shoots.
Objectives
The present study is part of a project undertaken to apply in vitro methods for the conservation of threatened species. This paper describes micropropagation and cold storage of Saussurea lappa.
Materials
The seeds of Saussurea lappa C.B.Clarke, obtained through the courtesy of the Lahul Kuth Growers Coop Marketing Society Ltd., Manali, Distt. Kulu, were used to raise aseptic seedlings which served as the source of explants.
Explant
The cotyledonary leaves, and hypocotyl and root segments derived from 15-day-old aseptic seedlings, were used as the explants
Initial culture
MS supplemented with 5 uM BAP and 3 mM GA3
Shoot multiplication
For micropropagation, shoot cultures were initiated from 10-15 mm long terminal portion of the hypocotyl segments bearing the plumule, and shoot buds produced in cotyledon cultures. At the end of each passage of shoot multiplication, individual shoots were separated, and transferred to fresh medium after clipping the leaves about 1 cm above the base of the shoots.
Rooting
The shoots multiplied on MS containing 5 uM BAP and 3 uM GA 3 were used for rooting.