Potential of In Vitro Culture of Scutellaria baicalensis in the Formation of Genetic Variation Confirmed by ScoT Markers

Abstract

The in vitro culture technique can be used for micropropagation of medicinal plants as well as for creating genotypes with an improved profile of phytochemical compounds. For this purpose, somaclonal variability may be used for the induction of genetic diversity among regenerants. The paper presents a protocol for obtaining Scutellaria baicalensis regenerants by indirect organogenesis and the assessment of their genetic variability with the use of start codon-targeted markers. The most intense process of indirect shoot organogenesis was observed on Murashige and Skoog medium supplemented with kinetin and 6-Benzylaminopurine (0.5 mg × dm−3 each)—7.4 shoot per explant on average. The callogenesis process occurred on the medium supplemented with TDZ, while the medium supplemented with GA3 allowed for direct shoot organogenesis and was used for the micropropagation of regenerants. In the analysis of plantlets obtained by indirect organogenesis, 11 ScoT markers generated a total of 130 amplicons, 45 of which were polymorphic. This analysis showed genetic diversity of regenerants in relation to the donor plant as well as within them, with mean similarity among the analyzed genotypes at the level of 0.90. This study confirms that the use of in vitro cultures allows for the possibility to generate genetic variability in Scutellaria baicalensis, which can be effectively revealed with the use of the SCoT marker.

The Influence of Cytokinin on the Multiplication Efficiency and Genetic Stability of Scutellaria baicalensis Regenerants in In Vitro Culture Conditions

“…The plant came from Experimental Farm of the University of Life Sciences in Lublin. Preparation of plant material to start in vitro culture was carried out according to the methodology provided by Gawroński and Dyduch-Siemińska [38] with modifications. Briefly, shoots were washed under tap water for an hour, rinsed for 15 min in distilled water, then sterilized with 70% ethanol for 15 s and 1% sodium hypochlorite for 5 min; they were subsequently washed three times (5 min each) in sterile distilled water.…”