Abstract
Linnaea borealis L. (Twinflower)—a dwarf shrub in the Linnaeeae tribe of Caprifoliaceae family—is distributed across the Northern Hemisphere. By means of this study, a reliable protocol for efficient micropropagation of uniform L. borealis L. var. borealis plantlets has been provided for the first time; callus culture was also established. Different initial explants, types of cultures, media systems, and plant growth regulators in Murashige and Skoog (MS) media were tested. Agitated shoot cultures in the liquid media turned out to be the best system for the production of sustainable plant biomass. After stabilization of the callus lines, the highest growth index (c.a. 526%) was gained for callus maintained on MS enriched with picloram. TLC and UHPLC-HESI-HRMS analysis confirmed the presence of phenolic acids and flavonoids, and for the first time, the presence of iridoids and triterpenoid saponins in this species. Multiplication of L. borealis shoot culture provides renewable raw material, allowing for the assessment of the phytochemical profile, and, in the future, for the quantitative analyses and the studies of the biological activity of extracts, fractions, or isolated compounds. This is the first report on in vitro cultures of traditionally used L. borealis rare taxon and its biosynthetic potential.
Highlights
Linnaea borealis L. (Twinflower), a creeping dwarf shrub, was Linnaeus’s favorite plant and became his symbol
The highest number of new shoots (7.63 ± 0.73) was obtained from double shoots on the solid medium supplemented with BAP (2.0 mg/L), IAA (0.1 mg/L) and GA3 (1.0 mg/L), as well as from stem fragments with nodes cultured on the solid Murashige and Skoog (MS) medium supplemented with BAP (1.5 mg/L), IAA (0.1 mg/L) and GA3 (1.0 mg/L), and MS with Kin (2.0 mg/L), IAA (0.1 mg/L), and GA3 (1.0 mg/L)—6.13 ± 0.64
This study demonstrated the influence of the type of the plant explant, hormonal supplementation in the medium and culture system on shoot multiplication and root development
Summary
Linnaea borealis L. (Twinflower), a creeping dwarf shrub, was Linnaeus’s favorite plant and became his symbol. (Twinflower), a creeping dwarf shrub, was Linnaeus’s favorite plant and became his symbol. This taxon was formerly assigned to the Caprifoliaceae family; it was transferred to the family of Linnaeaceae [1,2]. L. borealis is a small creeping evergreen perennial plant with the nature of a dwarf shrub, growing mainly in open pine woodlands [7]. Twinflower is rarely propagated generatively—seeds are often not produced or germination does not occur. L. borealis plants often intensively spread in forest stands by above-ground runners known as stolons, which consist of two types of stems: flowering and assimilation shoots. Stolons produce branches, forming large clonal patches consisting of groups of plants, which are genetically identical. Micropropagation may be a tool adopted to help propagation of the valuable species [3,7,9,10]
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