Hiraoka N et al, Plant Cell Reports 3:85-87 (1984)
Summary
Shoot cultures of Atractylodes lancea DC. (Compositae) have been established by inoculating the flower bud on Linsmaier-Skoog's medium supplemented with 10-5M naphthaleneacetic acid and l0-5M 6-benzylaminopurine. Shoots were multiplied on a medium containing 10-6M 6-benzylaminopurine. Propagated shoots rooted on a medium devoid of any plant growth regulators were transferred to potting soil and finally to the field.
Objectives
The rhizome of Atractylodes lancea DC. (Compositae) is one of the crude drugs prescribed clinically for diuretic, stomachic and sudorific purposes in traditional Chinese medicine. Since the plant bears no seeds in Japan, division of rhizomes is a usual procedure for its propagation. Only a few pieces of the rhizome ready for transplantation can be obtained from a 3- to 4-year-old plant by this method. Recently, Shoyama et al. (1983) reported the clonal multiplication of this plant through meristem culture. We have attempted independently to establish a procedure for rapid clonal propagation, starting from flower buds. This paper describes the results of these experiments.
Materials
Flower buds ( 10 mm long, 3 mm in diameter) were collected late in July or early in August from plants of about one month before the first flowering.
Explant
Flower buds ( 10 mm long, 3 mm in diameter)
Initial culture
The 40 bud segments were placed on LS containing one of four combinations of NAA and BAP at concentrations of 10-6M and/or 10-5M. After one month of incubation, a single shoot was induced from the explants on the medium supplemented with 10-5M NAA and 10-5M BAP.
Shoot multiplication
The effect of various kinds of cytokinins on shoot multiplication during four successive subcultures was investigated. Each one shoot explant consistently produced an additional shoot during the 4-week incubation period due to the addition of 10-6M BAP to the basal medium. The other cytokinins, kinetin, 6-isopentenylaminopurine or zeatin, were inferior to BAP in shoot multiplication. Therefore, we chose LS + 10-6M BAP as the optimal shoot-multiplication medium.
Rooting
For initiation of roots, multiple shoots were transferred to LS media of x l, x 2 I and x 4 dilution containing no growth regulators,
10-6M indoleacetic acid or NAA and 10-5M indoleacetic acid or NAA.　Considering these two factors and the percentage of rooted-shoots,
we chose LS of the full strength containing no growth regulators as a rooting medium.
Acclimation
Planting
The rooted cultures were transplanted in the following potting mixtures
: vermiculite only, a 6:2:2 vermiculite:sand:peat containing 1% slaked lime, 7:3 soil:sterile leaf mold or 4:3:3 soil:sand:peat containing 1% slaked lime mixture.