Methodological aspects of increasing the intensity of callusogenesis and organogenesis of Linum usitatissimum L. in vitro

Keywords: flax, in vitro, phytohormones, callus, organogenesis

Abstract

Aim. Improving methods for increasing the efficiency of obtaining callus cultures and somaclones of flax (Linum usitatissimum L.) in vitro. Methods. Hypocotyl segments were cultured on Murashige and Skoog nutrient medium supplemented with sucrose (30 g/l) and phytohormones at various concentrations. Other conditions: photoperiod 16 hours, illuminance 2500 lx, relative humidity 60–80%, air temperature 22–24°C. Results. The ability to form callus and somatic embryogenesis of flax depends on the phytohormonal composition of the nutrient medium, the size of the explants and the distance between them. Conclusions. For intensive callus formation and somatic embryogenesis in vitro, the optimal concentrations of BAP (mg/l) can be expressed as 1.0 ≤ BAP ≤ 1.75; the optimal concentrations of BAP for the medium supplemented with NAA (0.05 mg/l) 0.5 ≤ BAP ≤ 2.0; the optimal concentration of NAA for the medium supplemented with BAP (1.0 mg/l) 0.025 ≤ NAA ≤ 0.150; and the optimal concentrations of IAA for the medium supplemented with BAP (1.0 mg/l) 0.05 ≤ IAA ≤ 0.50. Addition of 0.5 mg/l GA3 to the medium with NAA and BAP is effective. It is optimal to use hypocotyl explants 3–6 mm long and place them at a distance of 1.5–2.5 cm from each other. Organogenicity of callus is significantly reduced in the process of subculturing.

References

Blinstrubienė A., Burbulis N., Kuprienė R. Effect of genotype and medium composition on linseed (Linum usitatissimum) ovary culture. Biologia. 2011. Vol. 66 (3). P. 465–469. doi: 10.2478/s11756-011-0028-z.

Burbulis N., Blinstrubienė A., Masienė R., Jonytienė V. Influence of genotype, growth regulators and sucrose concentration on linseed (Linum usitatissimum L.) anther culture. J. Food, Agricult. Environ. 2012. Vol. 10 (3–4). P. 764–767. doi: 10.1234/4.2012.3509

Janowicz J., Niemann J., Wojciechowski A. The effect of growth regulators on the regeneration ability of flax (Linum usitatissimum L.) hypocotyl explants in in vitro culture. BioTechnologia. 2012. Vol. 93 (2). P. 135–138. doi: 10.5114/bta.2012.46578.

Siegień I., Adamczuk A., Wróblewska K. Light affects in vitro organogenesis of Linum usitatissimum L. and its cyanogenic potential. Acta Physiol Plant. 2013. Vol. 35 (3). P. 781–789. doi: 10.1007/s11738-012-1118-4.

Sakhare S.P., Mendhulkar V.D. Embryo excised callus induction and rhizogenesis in Linum usitatissimum L. Int. J. Pharm. Bio. Sci. 2016. Vol. 7 (3). P. 507–511.

Blinstrubienė A., Burbulis N., Masienė R. Genotypic and exogenous factors affecting linseed ovary culture. Zemdirbyste-Agriculture. 2017. Vol. 104 (3). P. 243–248. doi: 10.13080/z-a.2017.104.031.

Mishchenko S.V. Influence of 6-benzylaminopurine on intensity of callusogenesis and organogenesis of Linum usitatissimum L. under in vitro conditions. The Bulletin of Kharkiv National Agrarian University. Series Biology. 2019. Vol. 2 (47). P. 92–100. doi: 10.35550/vbio2019.02.092. [in Ukrainian]

Mishchenko S.V. Effect of 1-naphthylacetic and indol-3-acetics acid on the intensity of callusogenesis and organogenesis of Linum usitatissimum L. in vitro. Factors in Experimental Evolution of Organisms. 2021. Vol. 28. P. 100–105. doi: 10.7124/FEEO.v28.1383. [in Ukrainian]

Anjum S., Abbasi B.H., Hano C. Trends in accumulation of pharmacologically important antioxidant-secondary metabolites in callus cultures of Linum usitatissimum L. Plant Cell Tiss Organ Cult. 2017. Vol. 129 (1). P. 73–87. doi: 10.1007/s11240-016-1158-3.

Khan I., Khan M.A., Shehzad M.A. et al. Micropropagation and production of health promoting lignans in Linum usitatissimum. Plants. 2020. Vol. 9 (6), 728. P. 1–18. doi: 10.3390/plants9060728.

Zahir A., Nadeem M., Ahmad W. et al. Chemogenic silver nanoparticles enhance lignans and neolignans in cell suspension cultures of Linum usitatissimum L. Plant Cell Tiss Organ Cult. 2019. Vol. 136 (3). P. 589–596. doi: 10.1007/s11240-018-01539-6.

Yildiz M., Sağlik C., Telci C., Erkilich E. G. The effect of in vitro competition on shoot regeneration from hypocotyl explants of Linum usitatissimum. Turk. J. Bot. 2011. Vol. 35 (2). P. 211–218. doi: 10.3906/bot-1005-26.