In vitro culture of the Ural endemic Gypsophila uralensis Less. (Caryophyllaceae)
Abstract
The culture of loose light green callus tissue from seedlings Gypsophila uralensis Less. was obtained using Murashige-Skoog (MS) and Woody Plant Medium (WPM) with the addition of BA 1.0 mg / l + IAA 0.1 mg / l (where BA = 6-benzylaminopurine and IAA = Indolyl-3-acetic acid). Callus had a high morphogenic activity for four passages. The percentage of viable callus varied from 82 % to 94 %. The average number of fragments per callus was 6.2 ± 0.4 on MS and 4.5 ± 0.4 on WPM. With prolonged cultivation (more than eight weeks), the callus darkened and died. The transition from callus proliferation to organogenesis was noted when the MS medium changed to Silene cretacea Saratov (SCS) medium. Further induction of morphogenesis took place on SCS medium with a complex set of growth regulators (BAP 0.2 mg / l + IAA 0.5 mg / l + CIN 1.0 mg / l + HA 1.0 mg / l). Up to 90 % of the callus switched to the formation of adventitious shoots after four to six weeks of cultivation. 2 to 13 shoots developed with a height of up to 2 cm with 2-4 pairs of well-developed green leaves depending on the size of the callus. Rhizogenesis was observed only on the WPM nutrient medium with the addition of IAA auxins from 0.1 mg / l to 1.0 mg / l and IAA 0.2 mg / l + IBA 0.5 mg / l (where IBA = indolylbutyric acid). The beginning of the formation of roots was observed after three to four weeks. The proportion of rhizogenesis was 51–53 %, i. e. both an increase in the concentration of IAA and the addition of IMA to the medium didn`t lead to an increase in the number of microturns with roots. The possibility of obtaining regenerated plants in the callus culture of Gypsophila uralensis was shown.
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