OBTAINING CALLUS CULTURES OF COMMON MUGWORT (ARTEMISIA VULGARIS L.)

Abstract

Background. One of the alternative sources of obtaining biologically active substances is a culture of plant cells in vitro. The objective of this work is to introduce Artemisia vulgaris in the callus culture. The plant grows in Central Yakutia, known as a species with a high content of various biologically active substances used in traditional medicine and widely spread on a territory researched. Materials and methods. The phytomass of Artemisia vulgaris L. was collected during the expeditions on the territory of Amga’s region of the Republic of Sakha (Yakutia) in June-July 2016–2018. For introduction of Artemisia vulgaris L. into a callus culture, the actual leaves of sterile plants were used as explants. The plants were obtained via cultivating the seeds of a wild plant in controlled conditions. The cultivation was performed on a Murashige-Skoog nutrient medium with the use of two different growth regulators: 2,4-dichlorophenoxyacetic acid and kinetin, 6-benzylaminopurine and α-naphthylacetic acid. Morphological analysis of cells of the obtained calli was performed using a light microscope. The obtained primary calli were selected for further transplantation on a Murashige-Skoog nutrient medium with different concentrations of growth regulators. The growth dynamics of the callus wet mass was studied during one cycle (21 days). Results. The biomass of primary callus Artemisia vulgaris L. in the Murashige- Skoog nutrient medium with supplementation of 2,4-dichlorophenoxyacetic acid and kinetin was light yellow in color and had a dense consistency, and with the addition of 6-benzylaminopurine and α-naphthylacetic acid, it was also pale yellow in color, but with easily separable loose structure. Morphological analysis of cells of primary callus obtained showed the predominance of round-ovoid cells with a distinct nucleus. In a nutrient medium supplemented with 2,4-dichlorophenoxyacetic acid, kinetin, and α-naphthylacetic acid, the peak increase in callus wet mass was observed on day 11 after transplantation. On a nutrient medium supplemented with 2,4-dichlorophenoxyacetic acid, α-naphthylacetic acid, and 6-benzylaminopurine the peak increase in the callus wet mass was observed on the 14th day. Conclusions. The primary callus of Artemisia vulgaris L. were obtained from leaf explants taken from sterile plants. A nutrient medium with growth regulators was optimized to obtain callus and for a further introduction of Artemisia vulgaris in the callus culture. On a nutrient medium supplemented with 2,4-dichlorophenoxyacetic acid, α-naphthylacetic acid, and 6-benzylaminopurine callus biomass of Artemisia vulgaris L. is three times heavier than the biomass of callus obtained on a nutrient medium with 2,4-D, kinetin, and NAA. The obtained callus cultures had a soft, easily separable, loose structure, pale yellow in color.

Keywords

• artemisia vulgaris l.
• callus culture
• ms nutrient medium
• callogenesis
• biomass
• growth curve