Russian and Foreign Cultivars of Honeysuckle (Lonicera edulis Turcz.): cultivation studies in vitro

Introduction. The demand for honeysuckle berries and planting material is growing. Clonal micropropagation is the most effective method for industrial plantations. The research objective was to study the effect of cytokinins and auxins on Russian and Canadian honeysuckle microshoots and roots. Study objects and methods. The study featured regenerated honeysuckle (Lonicera edulis Turcz.) of three Russian cultivars (Bakcharsky Velikan, Doch Velikana, Yugana) and two Canadian cultivars (Boreal Beauty, Boreal Beast). The experiment focused on the effect of sterilizing agents and sterilization time on the viability of honeysuckle explants at the stage of culture introduction in vitro. The effect of the growth regulator Cytodef in the QL nutrient medium on organogenesis was studied at the stage of micropropagation proper, the effect of auxin IBA on plant root formation – at the stage of rooting in vitro. Results and discussion. The greatest viability of honeysuckle explants (80–94%) was registered in the samples affected by Lizoformin 3000 (5%) and silver nitrate (0.2%) as sterilizing agents with a sterilization time of 10 min at the stage of in vitro culture introduction. The biggest quantity (8.8 pcs.) and total length (40.1 cm) of microshoots were observed when the content of cytokinin Cytodef in the culture medium QL was 0.3 mg/L at the stage micropropagation proper. The Boreal Beast cultivar had the largest total length of shoots (29.0 cm). The biggest quantity (5.5 pcs.) and total length (30.8 cm) of roots resulted from 0.5 mg/L of auxin IBA at the stage of rooting in vitro. Coconut substrate produced the highest survival rate (92–99%) at the stage of adaptation to non-sterile conditions in vivo, with the greatest number of leaves (8.1–10.2 pcs.) observed in Canadian cultivars. Conclusion. Cytodef and IBA proved to be effective growth-regulating substances for microplants of Russian and Canadian honeysuckle cultivars in vitro, which makes them promising for berry plantations.

Variability in the Chemical Composition of a New Aromatic Plant Artemisia balchanorum in Southern Russia

Lemon wormwood Artemisia balchanorum was recently introduced to southern Russia as a new aromatic plant. Based on biological and chemical characteristics, several populations with dominant citral, linalool, and geraniol production were selected for further development and maintained by seed propagation. Chemical analysis of five outstanding populations at three stages of annual development: vegetative, flower buds, and full flowering, confirmed that the seed populations retain the distinct dynamics of the dominant and minor components during the annual cycle and can be used for the commercial production of citral, linalool, and geraniol. Micropropagation in vitro allows for efficient clonal micropropagation and mass reproduction of elite cultivars and promising forms of A. balchanorum on a commercial scale but cannot serve as a source of direct and efficient production of secondary metabolites.

Producing Planting Material of Rubus arcticus L. by Clonal Micropropagation

The article presents the results of experimental studies on various propagation methods of the Arctic bramble in the Kostroma region. Industrial cultivation of forest berry plantations is a possible effective solution to the problem of low profitability of using nontimber forest products, reduction of the wild berries resources and their productivity and quality, and reclamation of cutover peatlands. It is advisable to use high-yield varietal planting material to create such plantations. Special attention is paid to propagation and production of healthy planting material using culture of plant cells and tissues. Data on sterilization of explants when introduced in vitro are given. The highest efficiency of sterilization was observed when using a chlorine-free eco-sterilizer (the plant survival rate on the MS nutrient medium was 90–93 %). The effect analysis of the passage number of regenerated plants on the multiplication factor of the Arctic bramble varieties was carried out. The optimal concentrations of cytokinins at the stage of micropropagation are shown. The largest number of the Arctic bramble roots was observed when adding 1.0 mg/L of Indole-3-butyric acid (IBA) and 0.5 mg/L of Ecogel to the nutrient medium. The technological and agrotechnical operations performed during the cultivation of planting material of forest berry plantations are considered. Data on the coefficients of vegetative propagation of plants and their resistance to diseases, yielding capacity, and recultivation of cutover peatlands are given. The best planting material of the Arctic bramble are ball-rooted seedlings. Sawdust and sphagnum were used in the cultivation of this berry on the peatland. The economic efficiency of its cultivation with the method of clonal micropropagation was 358.2 %. For citation: Makarov S.S., Tyak G.V., Kuznetsova I.B., Chudetsky A.I., Tsaregradskaya S.Yu. Producing Planting Material of Rubus arcticus L. by Clonal Micropropagation. Lesnoy Zhurnal [Russian Forestry Journal], 2021, no. 6, pp. 89–99. DOI: 10.37482/0536-1036-2021-6-89-99

Choice of explant, its size and method of its placement in a test tube to increase the productivity of clonal micropropagation of Fioletoviy ranniy variety

The paper is devoted to the optimization of one of the factors influencing the success of micropropagation – the choice of the optimal part of vine to take explant, size of the explant and the method of its exposure in a test tube. The studies have confirmed that the place, where micro-cutting was taken, its size and its orientation during planting can affect the speed of growth processes and the survival rate of plants. The experiment was put on the Purple Early grapevine variety. The part of vine for taking micro-cutting, best size of explant and the method of its orientation in the test tube are established.

Optimization of grapevine clonal micropropagation

Clonal micro-propagation ensures the production of genetically homogeneous, healthy virus-free planting material. This kind of propagation is influenced by genetic, physiological, hormonal, physical factors, and plant genotype. To increase the success of the process, it is necessary to optimize the methods of clonal micropropagation. The technology of clonal micropropagation of grapevine using a culture of apical meristems with a size of 0.1-0.2 mm was developed, the scheme of plant regeneration was improved, a new biotechnological techniques were developed for all stages of propagation. A method of processing of meristems with an ultrahigh frequency (microwave) electromagnetic field (EMF) in combination with a narrow-band laser was developed. Method for improving recovery from viral and bacterial infections using the growth regulator Emistim, Salicylic acid, the antibiotics Gentamicin and Cefotaxime was found. A method of water therapy was developed. The optimal parameters of intensity and duration of illumination were established. Ways of adaptation of healthy plants to non-sterile environmental conditions were optimised. Methods of testing of viral infection was improved. Techniques of planting healthy seedlings in film, greenhouses and open ground were developed. The creation of unique basic vine nursery in Ust-Donetsk region was the result of conducted studies.

Optimization of in vitro rooting and ex vitro adaptation conditions of Melissa officinalis L. microshoots during clonal micropropagation

Melissa officinalis L. is a perennial herbaceous essential and medicinal plant widely used in pharmacology, perfumery and cosmetics, as well as in alcoholic beverage and food industries. The low content of essential oil in lemon balm raw material determines the selection work aimed at creating high-oil cultivars. The use of clonal micropropagation method in vitro will increase the efficiency of this process and accelerate promising breeding samples multiplication. The aim of our research was to study the influence of cultivation conditions and cultivar on the M. officinalis in vitro rhizogenesis and ex vitro adaptation. It was found that the maximum frequency of shoot rooting (up to 93.3%) in cultivars ‘Citronella’ and ‘Sobornaya’ was on MS culture medium supplemented with 0.5 mg/l NAA. The number of roots was 10.1 and 13.6 pcs. per shoot, respectively. The highest rates of root formation for the cv. ‘Crimchanka’ was found on a culture medium supplemented with 1.0 mg/l IAA (8.7 roots per shoot). The mixture of peat, sand and perlite (2:1:2) as a substrate provided up to 93% of adapted ex vitro lemon balm microplants. The presented studies were used to develop a technique for clonal micropropagation of M. officinalis.

Influence of culture medium composition on the micropropagation of Thymus tauricus Klokov et Des.-Shost.

Thymus tauricus Klokov et Des.-Shost. is a perennial dwarf shrub of the Lamiaceae family. This plant has anti-inflammatory, antispasmodic, analgesic and antiseptic properties. The use of biotechnological methods makes it possible to increase the efficiency of traditional methods of breeding and seed production. Our investigation aimed to study the influence of the culture medium composition on the explants morphogenesis at 1-2th stages of Thymus tauricus clonal micropropagation. When comparing three cytokinins, the best explants development was revealed on culture media containing kinetin. Both high vitrification rate of microshoots (31.2-90.2%) and formation of small shoots were observed on media supplemented with BAP or TDZ. The most effective culture medium at the introduction stage is MS with 1.0 mg/l kinetin and 1.0 mg/l GA3 or 1.0 mg/l kinetin. On this medium, on average, 8.4-10.1 microshoots per explant and shoots length of 1.9 cm were obtained. It was found that the optimal culture medium at the actually propagation stage is MS with 1.0 mg/l Kin or 1.0 mg/l kinetin and 0.5 mg/l IAA, on which the multiplication index 26.9-28.5 were obtained. Induction of rhizogenesis with a frequency of 30.2-94.4% and a root length of 1.6-3.0 cm was noted on culture media for the second stage of clonal micropropagation. The results of the studies are the basis for development T. tauricus clonal micropropagation method.

Biotechnology methods to produce planting material of the genus Rubus L.

The work is devoted to improving clonal micropropagation methods of the genus Rubus representatives. When cultivating R. arcticus L. cultivars, the optimal concentration of 6-benzylaminopurine (6-BAP) was 0.3 mg L−1. Significant effect of FeEDTA concentration in the nutrient medium on multiplication of R. idaeus L. cultivars was established. The interaction of auxin type and plant genotype was revealed during rooting of blackberry cultivars and raspberry-blackberry hybrids. The nutrient medium supplemented with 0.5 mg L-1 indoleacetic acid contributed to the highest percentage of rhizogenesis in R. arcticus representatives (82%). The optimal growth regulator for conservation of raspberry explants was 6-BAP at a concentration of 0.3 mg L-1. One of the most representative in vitro collections of Rubus cultivars has been created.

Automation and digitalization of healthy fruit and berry plants reproduction

High-quality planting material is a condition for increasing productivity and labor productivity in the cultivation of agricultural products. High-quality planting material is material that is free from harmful pathogens. The process of plant healing requires significant time and material costs. However, with the proper organization, plant healing can be significantly accelerated and cheapened. The main condition for accelerating this process is maximum automation and digitalization of all stages of clonal micropropagation. The article shows the results of many years of work on the cultivation of healthy planting material. Variants of possible improvement of planting stock recovery are proposed with the introduction of the necessary hardware and software at specific stages of recovery and reproduction.

Improvement of clonal micropropagation technique of promising Lonicera caerulea L. cultivars

The work is focused on enhancement of in vitro propagation technique of valuable Lonicera caerulea L. cultivars. The impact of some nutrient medium components at main cultivation stages of the genus Lonicera L. representatives was studied. Some cultivars showed the positive effect of substituting sucrose for glucose in the medium at the multiplication stage (Diana (30 or 40 g L−1), Moskovskaya 23 (20 g L−1) and Yugana (40 g L−1)). When studying the influence of different auxin types on honeysuckle rhizogenesis, Solovey and Yugana preferred addition of 1.0 mg L−1 IBA into the medium. During the research of influence of different auxin concentrations on L. caerulea rooting the increased number of rooted microshoots of Goluboy Desert was demonstrated with the increasing of IAA concentration (from 0.5 to 3.0 mg L−1). Meanwhile, Diana and Yugana better rooted on the medium containing 1.0 mg L−1 IAA but Zolushka reached its highest rooting percentage on the medium supplemented with 0.5 mg L−1 IAA. The assessment of second-year plants undergone the whole cycle of clonal micropropagation was carried out 85% of these plants began to bear fruits.