Saposhnikovia divaricata (Apiaceae) – important species of Oriental Traditional Medicine: current research status, cultivation experience and biotechnological potential (a review)

Saposhnikovia divaricata (Turcz.) Schischk, originating from China, was studied for acclimatization by the National Institute of Medicinal Materials from 2018 to 2020. Its productivity and quality were evaluated, showing a total content of prim-O-glucosylcimifugin and 5-O-methylvisamminoside at 0.65%. Based on the research and selection conducted at the Tam Dao Medicinal Plant Research Station, National Institute of Medicinal Materials, using the mass selection method and quality evaluation of Saposhnikovia divaricata by High-Performance Liquid Chromatography with UV Detection (HPLC-UV), a variety with the following characteristics was selected: Herbaceous perennial plant, height 50–70 cm, with 2–3 pinnately compound leaves, serrated leaf edges, and dark green color. Inflorescences consist of many compound umbels, each having 5–7 smaller umbels, and each small umbel containing 4–9 white flowers. Schizocarps, brown when mature, oval-shaped, with a thousand-seed weight of approximately 5.83 g. The medicinal part is the taproot, cylindrical in shape, averaging 23.8 cm in length, with a yellow-brown color. This variety was evaluated for resistance to soft rot disease, with a severity level of 5 points (CIP scale), average yield 966.7 kg/ha. The total content of prim-O-glucosylcimifugin and 5-Omethylvisamminoside reached 1.0%.

The data mining analysis of the medication rule and the curative effect of traditional Chinese medicine in treating allergic rhinitis in children was performed by using the association rule Apriori algorithm. The model of interest degree was introduced to improve the Apriori algorithm, and the performance difference of the algorithm before and after improvement was analyzed. Traditional Chinese medicine prescriptions for the treatment of allergic rhinitis in children were selected from the dictionary of Chinese medicine formulations. The frequency, frequent itemsets, and the improved Apriori algorithm of each prescription were analyzed comprehensively. The results showed that both the execution time of the improved Apriori algorithm and the number of mining association rules were signally lower. 102 Chinese herbal compounds were selected, in which the occurrence frequency of Flos magnoliae was the highest (67 times, 5.33%). The occurrence frequency of diaphoretic drugs was the highest (412 times, 32.78%) in drug types. The occurrence frequency of Yu Ping Feng powder was the highest (21 times, 20.59%) in the Chinese herbal compound. After the association rule analysis of the improved Apriori algorithm, Perilla frutescens, Saposhnikovia divaricata, ginseng, Notopterygium root, and Astragalus propinquus Schischkin were often mixed with liquorice, and Flos magnoliae were usually mixed with Fructus xanthii and black plum. Compared with the conditions before treatment, the sign scores of children with allergic rhinitis were remarkably decreased after treatment with traditional Chinese medicine compounds (P < 0.05). The mining performance of the Apriori algorithm was improved by introducing an interest-based model. The treatment of traditional Chinese medicine on allergic rhinitis in children was combined with children's physiological and pathological characteristics of children, which used mild medicines.

Our representative studies to achieve sustainable use of crude drugs and ensure their stable quality are introduced: comprehensive studies on genetic, chemical, and sometimes pharmacological diversity of Asian medicinal plants including Paeonia lactiflora, Glycyrrhiza uralensis, Ephedra spp., Saposhnikovia divaricata, and Curcuma spp., as well as their related crude drugs. (1) For peony root, after genetic and chemical diversity analysis of crude drug samples including white and red peony root in China, the value-added resources with quality similar to red peony root were explored among 61 horticultural P. lactiflora varieties, and two varieties were identified. In addition, an optimized post-harvest processing method, which resulted in high contents of the main active components in the produced root, was developed to promote cultivation and production of brand peony root. (2) Alternative resources of glycyrrhiza, ephedra herb and saposhnikovia root and rhizome of Japanese Pharmacopoeia grade were discovered in eastern Mongolia after field investigation and quality assessment comparing Mongolian plants with Chinese crude drugs. Simultaneously, suitable specimens and prospective regions for cultivation were proposed. (3) Because of the wide distribution and morphological similarities of Curcuma species, classification of some species is debated, which leads to confusion in the use of Curcuma crude drugs. Molecular analyses of the intron length polymorphism (ILP) markers in genes encoding diketide-CoA synthase (DCS) and curcumin synthase (CURS) and trnK sequences, combined with essential oils analysis, were demonstrated as useful for standardization of Curcuma crude drugs. The above studies, representing various facets, can be applied to other crude drugs.

Abstract—Saposhnikovia divaricata (Turcz. ex Ledeb.) Schischk. (Apiaceae) is a useful medicinal plant, which contain a number of substances with a wide range of pharmacological activity; chromones, in particular, are found in the roots. For this reason, plants are heavily harvested from the wild, resulting in the population low density. S. divaricata is a taproot perennial monocarpic that reproduces by seeds only. The introduction of the species, the study of seed productivity and seed reproduction is a vital task for the development of industrial plant cultivation and the restoration of natural populations. The purpose of this work is to study seed production and biology of seed propagation. The generative shoot of S. divaricata is a synflorescence (a panicle of double umbels) with a floral unit being a double umbel. Plants from natural habitat and cultivated ones were studied. A comparative analysis of the seed production of double umbels, depending on their position on the rachis was carried out. The proportion of seed set (seed number/ovule number) and fruit set (fruit number/flower number) in a simple umbel, the potential and real seed productivity of a simple umbel, double umbel, and an individual plant were determined. Fruit of S. divaricata is a cremocarp consisting of two single-seeded mericarps. The seeds have a thin spermoderm, so they were stored and germinated with the pericarp. Seeds (mericarps) for germination were collected from natural populations and cultivated plants considering their position on the rachis, and stored for eight months under laboratory conditions (+23–25 °C). The results of the experiment included data on the duration of the period from the beginning of the experiment to the seed germination, the duration of the germination period (from the beginning of germination), laboratory germination of seeds (%), germination energy (%), rate of germination energy (%). All cultivated plants were monocarpic. The diameter of the synflorescence reaches 124 cm; on average, about 70 double umbels with fruits are formed on it. It was found that in introduced plants, seeds from the branches of the third order make up more than 50% of the real seed production, and from the second and fourth order branches – 21 and 26%, respectively. In natural populations, seeds from the fourth order branches account for 45% of the real seed production, of the third – 30%, of the 2nd – 15%. In the introduced plants, the real seed production of the second generation was about 6000 seeds per individual, which is higher than that of plants in natural populations, where the seed set is 65%. The seeds exhibit non-deep dormancy, and the period before germination does not exceed 10 days. Seed germination is dynamic; more than 50% of the germinated seeds have sprouted within seven days. Laboratory germination was higher in the seeds from the natural population, than from the introduced plants. The highest laboratory germination in introduced plants was found in seeds from double umbels of the second or third order branches – 72–73%, and from the natural populations – in seeds from the shoots of the third or fourth order branches – more than 90%.

The medicinal plant raw material containing essential oils always attract the attention of scientists from various countries worldwide. The search for new sources of the plant raw material rich in biologically active substances, particularly volatile compounds, remains a relevant task in modern pharmaceutical science. Our focus has been drawn to Rosa damascena Mill. of the “Veselka” (“Rainbow”) variety grown in the natural environment of Zaporizhzhia; its essential oil exhibits anti-inflammatory, antimicrobial, and antiseptic effects. To obtain a valuable medicinal plant raw material from Rosa damascena with a high content of essential oil, it was introduced into in vitro culture to produce aseptic regenerants, followed by their reintroduction into the natural environment. Aim. To conduct a comparative microbiological study of essential oils from Rosa damascena of the “Veselka” variety grown via clonal micropropagation in in vitro culture and in the natural environment of Zaporizhzhia. Materials and methods. The study objects were essential oils obtained by water distillation from rose petals cultivated using the method of clonal micropropagation in in vitro culture (at the premises of the Educational and Scientific Medical Laboratory Center with a vivarium) and the natural environment of Zaporizhzhia. The study of the antimicrobial activity was conducted in the microbiological laboratory of the Department of Microbiology, Virology, and Immunology at Zaporizhzhia State Medical and Pharmaceutical University. Results and discussion. The essential oils from Rosa damascena grown via clonal micropropagation in in vitro culture and under natural conditions in Zaporizhzhia effectively inhibited the growth of E. coli (21.3 mm and 12.0 mm, respectively) and S. aureus (11.3 mm and 10.2 mm, respectively). The results of the study on C. albicans indicate a high antifungal activity in both essential oils; the mean inhibition zone diameter in the experiments using the oil from Rosa damascena cultivated in in vitro culture is less: 33.3 mm compared to 40 mm in the experiments with the oil from roses grown in the natural environment of Zaporizhzhia. Conclusions. The method of clonal micropropagation of Rosa damascena of the “Veselka” variety is effective for obtaining a large amount of planting material in a short time with the subsequent reintroduction into the natural environment to expand the raw material base of valuable medicinal plants with a high content of essential oils. The results of the study of the antimicrobial activity of the essential oil of Rosa damascena of the “Veselka” variety cultivated through clonal micropropagation in in vitro culture have exhibited a high antimicrobial activity and moderate antifungal activity compared to the essential oil from Rosa damascena grown in the natural environment of Zaporizhzhia. Therefore, Rosa damascena Mill. of the “Veselka” variety cultivated under in vitro conditions is a promising source of essential oil with a high antibacterial effect in order to create new herbal formulations.

Use and exchange of aquatic genetic resources in aquaculture: information relevant to access and benefit sharing

Hedysarum zundukii Peschkova is one of the Fabaceae endemics of the flora of the Lake Baikal west shore. Because of its very poor renewal by seed production and seedling appearance biotechnological method, clonal micropropagation has been elaborated in order to improve its chances of conservation. A protocol for clonal micropropagation, including introduction, propagation, rooting, acclimatization, field cultivation and prolonged cold storage has been elaborated. Half-dose MS salts, benzylaminopurine 1 mg/dm3 and 2% sucrose were optimal components of the medium for clonal micropropagation. Sucrose was the superior carbon source by comparison with glucose and maltose. It was found that some agar brands were better for propagation, whereas other ones were better for rooting. Transplants produced from acclimatized plantlets vegetated successfully in field conditions, but did not survived in Irkutsk after wintering. However, the same transplants planted into their natural population survived successfully. Micropropagated plantlets retained their ability for propagation in vitro after 10 -12 months of cold storage at 4℃ with illumination. It was concluded that clonal micropropagation may be used as an additional means for conservation of H. zundukii.

Anacardium humile Saint Hilaire is a tropical shrub native to the Cerradobiome. It's a fruit-bearing plant having biological, medical, and social value. Knowing how natural populations' genetic variability is arranged enables for the development of conservation and sustainable use plans for the species and its environment. Interest in genetic studies of natural populations that combine evolutionary biology and ecology has increased in recent years. This work aims to assess how natural populations of Anacardium humile, an important tropical shrub species native to the Cerrado biome, are genetically structured. Based on investigations of 242 adult plants, six microsatellite loci previously identified from Anacardium occidentale were utilised to evaluate the regional genetic organisation and genetic diversity of eight natural Anacardium humile populations. These communities have a high level of genetic variety (expected heterozygozygosity He = 0.710), according to the findings. With a mean of 0.142 (p = 0.001), the endogamy coefficient was positive and significant for most populations. Although the genetic divergence between groups was small (\(\theta\) = 0.075 and GST = 0.066), it was significant (p = 0.0001). The results show that genetic conservation measures for A. humile populations should be undertaken in order to keep this species alive.

Fritillaria camtschatcensiscan reproduce by means of both sexual reproduction and clonal multiplication. Despite prolific seed production, no seedlings have been found for several years in natural populations on Mt Hakusan. The purposes of this paper are to clarify: (i) whether population size is maintained mainly by clonal multiplication; and (ii) if this is the case, to what extent occasional seedling establishment affects population growth rate and population structure. Two permanent quadrats were placed in subalpine meadows in 1992 on Mt Hakusan. Plant size, location and reproductive states for all ramets in the quadrats were recorded every year. Projection matrices were created based on field census, and computer simulation experiments were performed.Fritillaria camtschatcensishad two types of flower, male flower and cosexual flower, and they were changeable. This is the first report on sex lability inFritillaria. Clonal growth was more closely correlated with life‐history stages, especially with sexual states than with plant size. The population growth rate,λ, was 1.006 for the Mizuyajiri population and 1.047 for the Nanryu population, respectively. Seedlings were found in 1996 for the first time. These facts indicate that populations ofF. camtschatcensison Mt Hakusan can usually be maintained by clonal multiplication. However, it is not yet certain whether seeds germinate every year or whether a flush of seedling emergence occurs once in every few years in natural populations. Computer simulation revealed that: (i) there was a critical germination rate above which population growth rate suddenly increased; and (ii) occasional seedling establishment could provide almost the same contribution to population growth rate as that of annual seedling establishment. These results suggest that population size can be maintained mainly by clonal multiplication, and the role of sexual reproduction lies beyond maintaining the population size inF. camtschatcensis.

Hydroponic technologies combined with clonal micropropagation have the potential for large-scale plant growth and production of secondary metabolites. The aim of this study was to obtain raw materials Iris spuria L. (Art And Soul) in hydroponics coupled with clonal micropropagation, and its primary pharmacognostic analysis.&#x0D; When receiving the raw material Iris spuria L. (Art And Soul) at the stage of micro-multiplication itself, the most optimal content in the nutrient medium was 2.5-5.0 µm BAP. For a more complete realization of morphogenetic potentials it is necessary to alternate media with the content of phytohormones and hormone-free. At the same time, l-glutamine and adenine sulfate in an amount of 100 mg/l (MS+100mg/l L-glutamine+100mg/l adenine sulfate) should be added to the hormone-free media. For the adaptation of regenerated plants to non-sterile conditions and in the cultivation of raw materials can be used universal three-tiered aeroponic system, developed by All-Russian Research Institute of Agricultural Biotechnology, Moscow (Russia) (Iu.Ts. Martirosian).&#x0D; The obtained biotechnological raw materials (grass) were identified by macroscopic (external) and microscopic (anatomical) features (in accordance with the requirements of the State Pharmacopoeia of the Russian Federation XIV. 2018). As a result of the qualitative analysis of raw materials for the content of the main groups of biologically active substances, the following were revealed: phenols, flavonoids, tannins, alkaloids, glycosides, xanthones. For the herb Iris spuria L. the method of quantitative determination of flavonoids in terms of quercetin is developed.&#x0D; The dependence of the accumulation of quercetin and tannins on the hormonal composition of nutrient media, which makes it possible for Iris spuria L. (Art And Soul) to regulate the accumulation of these polyphenols in the production of plant raw materials.

The hawthorn (Crataegus ambigua) is a valuable species used for landscaping, medicinal purposes, and breeding. However, the decline of its natural populations underscores the need for effective microclonal propagation and preservation methods. This study conducted a comparative analysis of seed sterilization methods and obtained sterile seeds and micromasses. The influence of growth regulators, such as 6-BA, GA₃, IBA, and IAA, on microclonal propagation and rhizogenesis was also examined. Optimizing the conditions for slow growth is important for medium-term storage and the sustainable use of this species. These results can be used to develop effective protocols for the clonal micropropagation of C. ambigua, which is important for its conservation and introduction into breeding and reintroduction programs.

This monograph presents the results of many years research devoted to the development of complex biotechnological systems of creation the new forms and clonal micropropagation of such essential oil plants as lavender (Lavandula angustifolia Mill.), sage (Salvia sclarea L.), coriander (Coriandrum sativum L.), fennel (Foeniculum vulgare Mill.), anise (Anisum vulgare Gaerth.), yarrow (Achillea spp.), essential oil rose (Rosa spp.) and geranium (Pelargonium spp.). These biotechnologies are based on the investigation of features of isolated organs, callus tissues cultivation and morphogenesis induction in vitro. Chapter I deals with the general information on the methods of cell engineering that allow creating new plant genotypes and propagating them in vitro. The following Chapters summarize biotechnological research carried out at the Research Institute of Agriculture of Crimea (the former the Institute of Essential Oil and Medicinal Plants), using aromatic plant species, mainly grown in southern Russia (lavender, coriander, sage, fennel, rose yarrow etc.). The conditions of callus cultures obtaining from different explants were optimized and peculiarities of exogenous and endogenous factors effects on the callusogenesis efficiency of studied essential oil plants were detected. The methods of plant regeneration from calluses of different passages for more than fifty cultivars and samples of lavender, sage, coriander, fennel, anise, geranium, yarrow have been developed. The peculiarities of genotype, hormonal composition of culture medium, type of explant and number of passage influence on the morphogenesis induction have been determined. The means, 6 which permit to increase the frequency of organogenesis and somatic embryogenesis of callus cultures and to regenerate the plants during 1-3 years, were revealed. Furthermore, the method for creating essential oil rose hybrids using embryo culture and clonal micropropagation in vitro was developed. The peculiarities of abiotic stress factors effect at different biotechnological objects (calluses with and without morphogenesis and zygotic embryos) have been revealed. For the first time, the selective systems in vitro for screening forms with resistance to salinity, osmotic and low temperature stress have been elaborated for lavender, sage, geranium, coriander. For lavender, geranium, yarrow, fennel, rose and sage, methods of clonal micropropagation have also been created. The influence of cultivar, medium composition, explant disposition on the shoot and number of subcultivations on the propagation efficiency was shown. Based on the analysis of the regenerants from lavender, sage, geranium, coriander, yarrow and fennel calluses, the somaclonal variability by morphology and economic valuable characteristics have been determined, and prospects for breeding forms have been revealed. One of the practical results of these scientific studies is a new cultivar of clary sage Selinzh, which was created using somaclonal variants.

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.

The relevance of research on the development of biotechnology for clonal micropropagation of plants of the family Lamiaceae Lindl. is determined by the need for mass production of healthy, pure-grade planting material for the establishment of industrial plantations and the expansion of areas of essential oil crops in Ukraine. The aim of the research was to develop biotechnological methods of clonal micropropagation of essential oil plants of the Lamiaceae family – Lavandula angustifolia Mill., Mentha x piperita L., Salvia officinalis L. and Monarda fistulosa L. The task of the research was to select optimal conditions for effective cultivation of plants of the Lamiaceae family at four stages of clonal micropropagation. The main methods of research: laboratory, field, analytical, mathematical and statistical. The optimal nutrient media for the induction of in vitro morphogenesis and the multiplication stage were determined based on the basic medium of Murashige and Skoog: for L. angustifolia supplemented with kinetin (1.0 mg/l) and gibberellic acid (1.0 mg/l), for M. x piperita – with 6-benzylaminopurine (1.0 mg/l) and gibberellic acid (0.1 mg/l), for S. officinalis – with 6-benzylaminopurine (1.0 mg/l) and IOLK (0.5 mg/l ), for M. fistulosa – 6-benzylaminopurine (1.0 mg/l) and β-indolyl-3-oleic acid (0.1 mg/l). At the stage of multiplication, it is advisable to carry out seven to eight cycles of cultivation. At the stage of rooting of microshoots, the most effective for all studied plant species was determined to be the Murashige and Skuga nutrient medium with a halved concentration of components, supplemented with β-indolyl-3-oleic acid (0.5 mg/l) and β-indolyl-3-acetic acid ( 0.5 mg/l). The optimal substrate for plant adaptation to in vivo conditions is peat: perlite in a volume ratio of 3:1. Incorporation of the developed biotechnology of clonal micropropagation into the seeding system of essential oil crops of the Lamiaceae family will allow to quickly obtain healthy pure-bred planting material and introduce new productive varieties into production

Introduction. The last decade saw a considerable increase in the demand for European cranberry planting material (Oxyccocus palustris Pers.) among consumers of non-timber forest products. Cranberry possesses high nutritional and medicinal value. Cultivars and hybrids of European cranberry prove extremely productive for plantation growth using the method of clonal micropropagation with revitalized planting material. Study objects and methods. The research featured European cranberry plants of the Dar Kostromy cultivar and its hybrid form 1-15-635. The study focused on the effect of various medications and growth regulators on the biometric profile of European cranberry and its adaptation to non-sterile conditions at all stages of in vivo clonal micropropagation. Results and discussion. During the introduction stage, the highest viability belonged to the explants treated with AgNO3 (95–96%) and Lizoformin 3000 (5%) as the main sterilizing solutions at a 10-min exposure and a 5% solution of Ecosterilizer (1:1) at a 20-min exposure (90–95%). During the micropropagation proper, the number, average length, and total growth of shoots increased as the concentration of cytokinin 2ip in the WPM 1/4 nutrient medium rose from 1.0 to 5.0 mg/L. At the stage of in vitro rooting, the maximal number, average length, and total growth of roots in regenerated plants for both cultivars were observed when Kornerost 5.0 mg/L was added to the WPM 1/4 nutrient medium. At the stage of adaptation to in vivo conditions, Micogel 0.2 mg/L contributed to the highest survival rate (94–100%). Conclusion. During clonal micropropagation in vitro, the biometric profile of European cranberry (Oxyccocus palustris Pers.) and its survival rate under non-sterile conditions in vivo proved to depend on various growth-regulating substances and their concentrations.