## 2 Materials and Methods
### Proliferation  
For proliferation, explants were placed in a medium containing either N-(Phenylmethyl)-7H-purin-6-amine (6-benzyl adenine (BA, 1.0 mg\(\cdot\)dm\({}^{-3}\)) or 6-(3-Hydroxybenzylamino)purine (meta-topolin (mt, 1.0 mg\(\cdot\)dm\({}^{-3}\)). The medium composition included 100% MS macronutrients--1.65 g\(\cdot\)dm\({}^{-3}\) NH\({}_{4}\)NO\({}_{3}\), 440 mg\(\cdot\)dm\({}^{-3}\) CaCl\({}_{2}\), 180.7 mg\(\cdot\)dm\({}^{-3}\) MgSO\({}_{4}\), 170 mg\(\cdot\)dm\({}^{-3}\) KH\({}_{2}\)PO\({}_{4}\) and 1.90 g\(\cdot\)dm\({}^{-3}\) KNO\({}_{5}\)--and 100% MS micronutrients--6.2 mg\(\cdot\)dm\({}^{-3}\) H\({}_{3}\)BO\({}_{3}\), 0.02 mg\(\cdot\)dm\({}^{-3}\) CoCl\({}_{2}\), 27.8 mg\(\cdot\)dm\({}^{-3}\) FeSO\({}_{4}\), 22.3 mg\(\cdot\)dm\({}^{-3}\) MnSO\({}_{4}\), 0.83 mg\(\cdot\)dm\({}^{-3}\) KI, 0.25 mg\(\cdot\)dm\({}^{-3}\) ZnSO\({}_{4}\) and 0.02 CuSO\({}_{4}\). The medium contained 100% MS vitamins--2 mg\(\cdot\)dm\({}^{-3}\) glicine, 0.1 mg\(\cdot\)dm\({}^{-3}\) pyridoxine, 0.1 mg\(\cdot\)dm\({}^{-3}\) niacin and 0.1 mg\(\cdot\)dm\({}^{-3}\) thiamine--and 100% WPM vitamins--2 mg\(\cdot\)dm\({}^{-3}\) glicine, 0.1 mg\(\cdot\)dm\({}^{-3}\) pyridoxine, 0.1 mg\(\cdot\)dm\({}^{-3}\) pyridoxine, 0.1 mg\(\cdot\)dm\({}^{-3}\) priacin and 1 mg\(\cdot\)dm\({}^{-3}\) thiamine. The medium contained 100% MS vitamins--2 mg\(\cdot\)dm\({}^{-3}\) glicine, 0.1 mg\(\cdot\)dm\({}^{-3}\) pyridoxine, 0.1 mg\(\cdot\)dm\({}^{-3}\) niacin and 1 mg\(\cdot\)dm\({}^{-3}\) thiamine. The growth hormone concentration was determined based on scientific reports of micropropagation of \(L\). _acerulea_ var. _kamtschatica_ \begin{table}
\begin{tabular}{c c c c} \hline \hline Stages & Initiation & Proliferation & Rooting \\ \hline Macronutrients & 100\% MS \({}^{1}\) & 100\% MS & 50\% MS \\ \hline Micronutrients & 100\% MS & 100\% MS & 50\% MS \\ \hline \multirow{2}{*}{Additional micronutrients} & Fe(III)EDDHA & Fe(III)EDDHA & Fe(III)EDDHA \\  & (10 mg\(\cdot\)dm\({}^{-3}\)) & (10 mg\(\cdot\)dm\({}^{-3}\)) & (5 mg\(\cdot\)dm\({}^{-3}\)) \\ \hline Vitamins & 100\% MS, & 100\% MS, & 200\% WPM \\  & 100\% WPM \({}^{2}\) & 100\% WPM & \\ \hline \multirow{2}{*}{Additional vitamins} & folic acid (0.5 mg\(\cdot\)dm\({}^{-3}\)) & folic acid (0.5 mg\(\cdot\)dm\({}^{-3}\)), & \\  & pantothenic acid (1 mg\(\cdot\)dm\({}^{-3}\)) & folic acid (0.5 mg\(\cdot\)dm\({}^{-3}\)) & \\ \hline Myo-inositol & 100 mg\(\cdot\)dm\({}^{-3}\) & 100 mg\(\cdot\)dm\({}^{-3}\) & 100 mg\(\cdot\)dm\({}^{-3}\) \\ \hline Sucrose & 20 g\(\cdot\)dm\({}^{-3}\) & 20 g\(\cdot\)dm\({}^{-3}\) & 15 g\(\cdot\)dm\({}^{-3}\) \\ \hline \multirow{2}{*}{Growth regulators} & 1 mg\(\cdot\)dm\({}^{-3}\) BA &
\begin{tabular}{c} 1 mg\(\cdot\)dm\({}^{-3}\) BA \\ or 1 mg\(\cdot\)dm\({}^{-3}\) mT \\ \end{tabular} & 2 mg\(\cdot\)dm\({}^{-3}\) IBA \\ \hline pH & 5.7 & 5.7 & 5.7 \\ \hline Agar & 8 g\(\cdot\)dm\({}^{-3}\) & 8 g\(\cdot\)dm\({}^{-3}\) & 8 g\(\cdot\)dm\({}^{-3}\) \\ \hline \hline \end{tabular} \({}^{1}\) MS—[PERSON] [28]. \({}^{2}\) WPM—[PERSON] et al. [29].  
\end{table} 
Table 1: Composition of media used for micropropagation of Japanese hascap. and results of preliminary experiments [8,30]. Each treatment was represented by at least 6 jars (\(\times\)8 explants). During proliferation at the end of passage, the numbers of short (3-9 mm) and long (>10 mm) shoots and the total number of and length of shoots were measured. In addition, the number of cultures that were dead or had apical necrosis was counted as well as the number of healthy shoots and the size of calluses in the cultures. Cultures were grown in 300-500 mL glass jars, filled with 50 mL of medium, closed with autoclavable, transparent lids. They were grown on shelves in a culture room with the temperature ranging from 26 \({}^{\circ}\)C during the day to 24 \({}^{\circ}\)C at night. The photoperiod was 16 h/8 h (day/night), with light of 26 umol\(\cdot\)m\({}^{-2}\cdot\)s\({}^{-1}\) PPFD provided by Grolux fluorescent tubes.