The influence of light quality and cytokinin content in media on growth, development, photosynthetic pigments and secondary metabolite content of Myrtus communis L. was evaluated in an in vitro culture. Various treatments with light emitting diodes (LEDs): 100% blue (B), a mix of 70% red and 30% blue (RB) and 100% red were applied and compared with a traditional fluorescent lamp as control. Axillary shoots were incubated on Murashige and Skoog medium with 30 g dm⁻³ sucrose, 0.5% BioAgar, 0.5 μM 1-naphthaleneacetic acid and different concentrations of 6-benzyladenine (BA): 1, 2.5 and 5 µM. Cultures were maintained for 6 weeks in 23/21 ± 1 °C (day/night), 80% relative humidity and 16/8 h photoperiod; photosynthetic photon flux density (PPFD) was 35 µmol m⁻² s⁻¹ in all treatments. Light spectra and BA content in media affected biometrical and phytochemical M. communis properties. Red LEDs and 5 µM BA resulted in the highest multiplication rate. The highest shoots were obtained under red LEDs, but with the lowest concentration of cytokinin in media. Fresh weight was greatest on LEDs containing blue light in the spectrum (B and RB); moreover, 5 µM BA increased dry weight. Photosynthetic pigment levels were lower under LED light compared to control lamps. Phenolic acids and flavonoids were identified in M. communis leaf extracts. Myricetin was the major constituent with highest concentration under red LEDs and highest BA level.

In vitro callus cultures of Verbena officinalis L. were maintained on solid Murashige and Skoog medium, enriched with 1 mg dm-3 BA and 1 mg dm-3 IBA under LED lights (red, blue, red/blue 70%/30%), in darkness and under control fluorescent lamps. The measurements of 2 phenylpropanoid glycosides (verbascoside and isoverbascoside) and 23 phenolic acids were performed in methanolic extracts from the biomass collected after 2-, 3- and 4-week growth cycles using the HPLC-DAD method. The presence of verbascoside, isoverbascoside and additionaly 7 phenolic acids (protocatechuic, chlorogenic, vanillic, caffeic, ferulic, o-coumaric and m-coumaric acids) was confirmed in all extracts. Blue and red/blue lights stimulated the accumulation of verbascoside (max. of 6716 and 6023 mg 100 g-1 DW after a 4-week growth cycle) and isoverbascoside (max. 333 and 379 mg 100 g-1 DW also after 4 weeks). The maximum amounts of verbascoside and isoverbascoside were respectively 1.8- and 7.0-fold higher than under the control conditions. Phenolic acids were accumulated in different amounts, and the maximum total amounts ranged from 36 to 65 mg 100 g-1 DW. LED lights also stimulated their accumulation in comparison with darkness and control. The main phenolic acids included: m-coumaric acid (max. 39 mg 100 g-1 DW), ferulic acid (max. 12 mg 100 g-1 DW), and protocatechuic acid (max. 13 mg 100 g-1 DW). Additionally, the quantities of photosynthetic pigments (chlorophyll a, b and carotenoids) were estimated in acetonic extracts using spectrophotometry. Red/blue light stimulated the biosynthesis of pigments (max. total content 287 μg g-1 FW after 4-week growth cycles). This is the first study describing the effect of LED lights on the production of phenylpropanoid glycosides and phenolic acids in V. officinalis callus cultures. Very high amounts of verbascoside and isoverbascoside are interesting from a practical point of view.

Light quality is essential in in vitro cultures for morphogenesis process. Light emitting diodes system (LED) allows adjustment as desired and the most appropriate light spectrum. The study analyzed the influence of different LED light quality on the balance of endogenous phytohormones and related compounds (PhRC) in in vitro multiplied axillary shoots of Gerbera jamesonii. Over a duration of 40 days, the shoots were exposed to 100% red light, 100% blue light, red and blue light at a 7:3 ratio with control fluorescent lamps. Every 10 days plant tissues were tested for their PhRC content with the use of an ultra-high performance liquid chromatography (UHPLC). Shoots’ morphometric features were analyzed after a multiplication cycle. We identified 35 PhRC including twelve cytokinins, seven auxins, nine gibberellins, and seven stress-related phytohormones. Compounds content varied from 0.00052 nmol/g to 168.15 nmol/g of dry weight (DW). The most abundant group were stress-related phytohormones (particularly benzoic and salicylic acids), and the least abundant were cytokinins (about 370 times smaller content). LED light did not disturb the endogenous phytohormone balance, and more effectively mitigated the stress experienced by in vitro grown plants than the fluorescent lamps. The stress was most effectively reduced under the red LED. Red and red:blue light lowered tissue auxin levels. Blue LED light lowered the shoot multiplication rate and their height, and induced the highest content of gibberellins at the last stage of the culture.

A mixture of red and blue light-emitting diodes (LEDs; at a ratio of 7:3, respectively) were used to analyze the effects of different photosynthetic photon flux densities (PPFDs) (40, 80, and 120 µmol m⁻² s⁻¹ hereafter known as LED 40, 80, and 120, respectively) on the micropropagation of Gerbera jamesonii Bolus shoots. The experiment also examined the effect of 6-benzyladenine (BA) in 1, 2.5, and 5 µM concentrations in the media. Biometrical observations and analyses of leaf morphometry and photosynthetic pigment content were conducted. Shoot multiplication increased with an increasing BA concentration. A PPFD of 80 µmol m⁻² s⁻¹ and 5 µM BA is suggested as efficient for shoot propagation and economically viable. LED 120 increased the leaf blade area and its width, and circularity and elongation ratios. The intensity of light did not affect the fresh weight, which increased at higher BA concentrations (2.5 and 5 μM). The dry weight content decreased with increasing cytokinin concentration; the greatest content was observed on media with 1 µM BA under PPFD 120 µmol m⁻² s⁻¹. LED 80 increased the photosynthetic pigments content in the leaves in comparison to the standard intensity of LED 40. Increased BA concentration raises the content of chlorophyll a.

The experiment evaluated the influence of different light qualities and 6-benzyladenine (BA) concentration in a medium on the leaf response of multiplied Gerbera jamesonii Bolus ex Hook. f.‘Big Apple’ shoots. Three different light-emitting diode (LED) spectra—100% blue (B), 100% red (R) and red and blue mixture (7:3, RB)—were used, and a fluorescent lamp was used as a control (Fl). Concentrations of BA in Murashige and Skoog (MS) medium were 1, 2.5 and 5 µM. Leaves developed under 100% blue light had a lower frequency of stomata and a smaller area as compared with those from plants exposed to light with red in spectrum. Under 100% red light, the leaf area and the frequency of stomata increased along with growing concentration of BA in the medium. The thickest mesophyll was spotted in the cross-section of leaves exposed to the blue LED light. Leaves developed under the 100% red light had the thinnest mesophyll layers. Increasing concentration of BA in the medium resulted in enhanced leaf blade thickness. The cross-section of leaf vascular bundles was only half of that in petioles. The leaves under the LED combinations had larger vascular bundles than those under fluorescent light. The highest level of photosynthetic pigments was noticed in the leaves grown under LED R and RB lights. Our study demonstrated that 2.5 µM BA and a mixture of blue and red light provided by LED improved leaf quality during multiplication of gerbera shoots.