Morphogenesis features of <i>Hedysarum alpinum</i> L. in <i>in vitro</i> culture

Abstract

The plants of Hedysarum alpinum L. in the North are characterized by a long pregenerative period, the laboriousness of seed propagation, low field germination of seeds, rare self-sowing, and the impossibility of vegetative propagation. In connection with this, it seemed relevant to introduce this rare species (included in the Red Book of the Komi Republic) into aseptic culture to preserve and maintain the gene pool, as well as to further develop the technology of its cultivation to obtain medicinal raw materials. The morphogenesis was studied, and the peculiarities of the regeneration of adventitious shoots of Hedysarum alpinum in in vitro culture were revealed. During three subcultivations, the influence of two cytokinin growth regulators, 6-benzylaminopurine (BAP) and kinetin (KIN), at concentrations from 0,5 to 1,0 mg/l, on the regeneration potential of different explants of Hedysarum alpinum in in vitro culture was determined on the WPM environment. A high frequency of regeneration of shoots from single-node segments and cotyledon nodes was obtained 79% and 65%, respectively. In the second and third passages, there was an increase in the regenerative activity of the cotyledon node explants and an increase in the multiplication factor to 5 shoots per explant. It was shown that relatively high concentrations (1,0 mg/l) of BAP and CIN increase the number of shoots per explant but inhibit their growth and development. For the reproduction stage proper, the medium containing BAP 0,5 + CIN 0,5 + IAA 0,1 mg/l is optimal. It was shown that the initiation of rhizogenesis actively took place on the WPM medium supplemented with IAA 0,5 and IBA 0,5 mg/l, on which the proportion of rhizogene explants was 77%. The maximum values for the length and number of roots were obtained on the medium with IAA 1,0 mg/l and with the combined use of two auxins (IAA with IBA at 0,5 mg/l). The regenerated plants were transplanted into non-sterile conditions, and their survival rate under ex vitro conditions was 50%.