In vitro propagation of Aconitum balfourii Stapf.: an important aconite of the Himalayan alpines

Small leaf segments, obtained from in vitro sprouted axillary buds, have been used to develop an efficient micropropagation protocol for Aconitum balfourii Stapf., an important medicinal herb of the Indian Himalayan alpines. Following the induction of callus from cultured leaf segments on Murashige and Skoog (1962; MS) medium containing 4.5 muM 6-benzyladenine (BA) and 26.9 muM alpha-naphthaleneacetic acid (NAA), the highest values for shoot induction were obtained on the same medium supplemented with the same concentration (4.5 muM) of BA and a lower concentration of NAA (1.4 muM). These individual shoots could be further multiplied easily, with maximum shoot proliferation obtained on MS medium containing 1.1 muM BA only. The shoots obtained either directly from the callus or after multiplication on the above medium, were found to root efficiently on MS medium supplemented with 12.3 muM indole-3-butyric acid (IBA). After in vitro hardening in the culture room, the plantlets were transplanted to a greenhouse where the growth of plants was normal. Comparison of mitotic chromosome numbers of root tip cells, protein profile and alkaloid content of tubers of in vitro raised and seed grown plants of comparable age showed equal number of chromosomes, identical protein profile and similar amounts of diterpenoid alkaloids (aconitine and pseudoaconitine). This paper is the first report describing a complete regeneration protocol, along with quantitative data, for A. balfourii using in vitro methods.