Kucharova Elena Valerievna, Leading engineer, Educational-Scientific Laboratory of Molecular Genetic and Cellular Technologies, Institute of Natural Sciences, North-Eastern Federal University in Yakutsk (48, Kulakovskogo street, Yakutsk, Russia), E-mail: firstname.lastname@example.org
Okhlopkova Zhanna Mikhaylovna, Candidate of biological sciences, associate professor, biological department, Institute of Natural Sciences, North-Eastern Federal University in Yakutsk (48, Kulakovskogo street, Yakutsk, Russia), E-mail: email@example.com
Antonova Elena Evgen'evna, Student, North-Eastern Federal University in Yakutsk (48, Kulakovskogo street, Yakutsk, Russia), E-mail: firstname.lastname@example.org
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.
1. Krasnoborov I. M. Flora Sibiri: v 14 t. [Siberian flora: in 14 volumes]. Novosibirsk: Nauka, 1997, vol. 13, 472 p. [In Russian]
2. Zakharova V. I. Raznoobrazie rastitel'nogo mira Yakutii [The diversity of the plant world of Yakutia]. Novosibirsk: Izd-vo SO RAN, 2005, 328 p. [In Russian]
3. Danilova N. S., Borisova S. Z., Ivanova N. S. Vestnik Severo-Vostochnogo federal'nogo universiteta imeni M. K. Ammosova [Bulletin of Noeth-Eastern Federal University in Yakutsk]. 2011, vol. 8, no. 1, pp. 11–17. [In Russian]
4. Ivanova N. S., Danilova N. S., Borisova S. Z. Dekorativnye rasteniya Yakutii: atlasopredelitel' [Ornamental plants of Yakutia: atlas-determinant]. Moscow: Fiton+, 2012, 248 p. [In Russian]
5. Zhigzhitzhapova S. V., Soktoeva T. E., Radnaeva L. D., Taraskin V. V., Namzalov B.-Ts. B. Vestnik Buryatskogo gosudarstvennogo universiteta [Bulletin of Banzarov Buryat State University]. 2012, no. S3, pp. 74–77. [In Russian]
6. Govindaraj S., Kumari B. D., Cioni P. L., Flamini G. Journal of bioscience and bioengineering. 2008, vol. 105 (3), pp. 176–183.
7. Fedorov A. A., Efremova M. I., Chirikova N. K. Fundamental'nye issledovaniya [Fundamental research]. 2014, no. 11-9, pp. 1981–1983. [In Russian]
8. Butenko R. G. Biologiya kletok vysshikh rasteniy in vitro i biotekhnologii na ikh osnove: ucheb. posobie [Biology of cells of higher plants in vitro and biotechnology based on them: teaching aid]. Moscow: FBK-PRESS, 1989, 160 p. [In Russian]
9. Borzabad R., Sudarshana M., Niranjan M. Modern Applied Science. 2010, vol. 4 (9). DOI 10.5539/mas.v4n9p130.
10. Kumar S., Kumari B. Asian Journal of Biotechnology. 2010, vol. 2 (1), pp. 37–45. DOI 10.3923/ajbkr.2010.37.45.
11. Hedayati N., Hosseini R., Hedayati B. Iranian Journal of Pharmaceutical Research. 2013, vol. 12, p. 684.