Chemical Composition of Essential Oils and Local Knowledge of Myrica gale in Lithuania.

Myrica gale L. (Myricaceae) is an essential oil-bearing plant that is rare in Lithuania and naturally grows only in the western part of the country. The aim of this study was to analyze the composition of essential oils of Myrica gale in different habitats in Lithuania and in different parts of the plant, as well as evaluate the local knowledge about M. gale as a medicinal and aromatic plant. Samples of fruits and leaves (from one and three M. gale populations, respectively) were studied separately. Essential oils were isolated from dried fruits and leaves by hydrodistillation and analyzed by GC/FID and GC/MS methods. Results showed that M. gale fruits accumulated 4.03 ± 2.13% essential oils, meanwhile leaves-up to 19 times less. A total of 85 compounds were identified in the essential oils of the M. gale. Monoterpene hydrocarbons accounted for about half of the total essential oil content; meanwhile, either monoterpene hydrocarbons or sesquiterpene hydrocarbons (depending on habitat) dominated in leaves. The main compounds (depending on habitat) in essential oils of fruits and leaves were α-pinene, 1,8-cineole, limonene, δ-cadinene, and (E)-nerolidol. The high variation in the composition of M. gale essential oils suggests the presence of different chemotypes within the studied habitats of this plant. Evaluation of local knowledge of M. gale through the survey of 74 residents of 15 villages in western Lithuania showed that only 7% of respondents knew this plant. Poor knowledge of M. gale could be related to the narrow range of the natural species' distribution in Lithuania.

On the Prospects of In Situ Conservation of Medicinal- and Aromatic-Plant Genetic Resources at Ancient-Hillfort Sites: A Case Study from Lithuania.

Twenty-three ancient-hillfort sites were investigated to evaluate the potential for the in situ conservation of medicinal- and aromatic-plant populations. An evaluation of the site's suitability was carried out by employing three major groups of criteria: species-specific, site-specific, and threat assessment. The species-specific criteria included the total species number, target species number, the cover-abundance of the target species estimated by mean Braun-Blanquet score, and, as an additional criterion, the number and cover-abundance of crop wild relatives. The site-specific criteria included site evaluation with respect to climatic region, the area size of a site, the habitat type, and the site's protection status. The threat assessment was focused on anthropogenic activities, such as recreational, agricultural, and others. The total number of vascular plant species inventoried was 264, including 82 species of medicinal and aromatic plants (MAP). There was a strong and highly significant correlation between the total and the MAP species numbers (rs = 0.77, p < 0.001), and the two most species-rich sites, Zuklijai and Pamiske, contained the highest total and MAP species numbers. The investigated hillfort sites covered the populations of 49 species, or about 33% of the priority species list, with 5 or more populations. The most frequent species, Hypericum perforatum, occurred at 21 sites. The twenty-three hillfort sites represent three of the four climatic regions and six of the ten climatic subregions of Lithuania. Although these hillfort sites are quite small (1.24 ± 0.75 ha on average, without buffer zone), they are scattered across the country and are state-protected as archaeological objects, which makes them suitable for the in situ conservation of MAP genetic resources. In addition, seven hillfort sites (30.4% of the investigated ones) belong to the European network of special areas of conservation of habitats (Natura 2000), thus increasing their international importance. The threat assessment showed that anthropogenic activities (recreational, agricultural, etc.) are among the major factors affecting target-species populations.

Development of a Methodology for Maintenance of Medicinal Plant Genetic Reserve Sites: A Case Study for Lithuania.

In the context of climate change, in situ conservation of plant genetic resources is becoming increasingly important as it supports natural (ecological and evolutionary) adaptations of plants to the changing environment. The aim of this study was to synthesize a comprehensive general methodology for the maintenance of genetic reserve sites of medicinal plants based on the analysis of relevant legal documents, literature sources, databases, and authors' own experience. A methodology was developed for the application of various maintenance measures for target species populations in genetic reserve sites to ensure their long-term sustainability. It uses a systematic approach to the intervention measures by grouping them into habitat-specific and factor-specific ones, and follows the specific principles of biodiversity conservation, such as the concept of ecosystem, priority of in situ conservation, caution and validity of decision-making, and regional approach. An extensive discussion on major intervention measures is provided. The methodology is intended to be used as a tool for the preparation and implementation of management plans of individual medicinal plant genetic reserve sites by the responsible agencies and protected area managers and is directly linked to the implementation of the EU (European Union) Biodiversity Strategy to 2030 at the national level.

Bioactive Compounds, Antioxidant, and Antibacterial Properties of Lonicera caerulea Berries: Evaluation of 11 Cultivars.

The aim of the study was to evaluate 11 cultivars of blue honeysuckle (Lonicera caerulea L.) for bioactive compounds, antioxidant capacity, and the antibacterial activity of berries. Total phenolic contents (TPCs) and total anthocyanin contents (TACs) were established by using ethanolic extracts. For contents of organic acids and saccharides, aqueous extracts were used, and vitamin C was determined by using oxalic acid solution. DPPH• radical scavenging capacity was evaluated by using ethanolic extracts; antibacterial activity was assessed by using both ethanolic and aqueous extracts. The TPC varied from 364.02 ± 0.41 mg/100 g in 'Vostorg' to 784.5 ± 0.3 mg/100 g in 'Obilnaja', and TAC ranged from 277.8 ± 1.1 mg/100 g in 'Celnocnaja' to 394.1 ± 8.4 mg/100 g in 'Nimfa'. Anthocyanins comprised 53.8% of total phenolic contents on average. Among organic acids, citric acid was predominant, averaging 769.41 ± 5.34 mg/100 g, with malic and quinic acids amounting to 289.90 ± 2.64 and 45.00 ± 0.37 mg/100 g on average, respectively. Contents of vitamin C were 34.26 ± 0.25 mg/100 g on average. Organic acids were most effective in the inhibition of both Gram-positive and Gram-negative bacteria tested. In conclusion, berries of L. caerulea are beneficial not only for fresh consumption, but also as a raw material or ingredients of foods with high health-promoting value.

Dynamics of Isomeric and Enantiomeric Fractions of Pinene in Essential Oil of Picea abies Annual Needles during Growing Season.

Norway spruce (Picea abies (L.) H. Karst.) is one of the most important commercial tree species distributed naturally in the Boreal and subalpine forest zone of Europe. All parts of spruce trees, including needles, accumulate essential oils with a variety of chemical properties and ecological functions, such as modulating plant-insect communication. Annual needle samples from 15 trees (five from each of three habitats) of 15-17 years old were assayed for essential oils and their major compounds, including α-pinene, ß-pinene, (1S)-(-)-α-pinene, and (1R)-(+)-α-pinene across a growing season. Results showed strong positive correlation between percentages of α- and ß-pinene isomers (r = 0.69, p < 0.05) and between pinene isomers and essential oils: α-pinene correlated with essential oil stronger (r = 0.62, p < 0.05) than ß-pinene (r = 0.33, p < 0.05). Correlation analyses performed with some weather conditions, including average monthly temperature, growing sum of effective temperatures over 5 °C, duration of sunshine, accumulated precipitation, relative humidity, and pressure, showed that temperature is the most important weather condition related to pinene dynamics: negative correlations of moderate strength were established between percentages of α- and ß- pinenes and average monthly temperatures (r = -0.36, p < 0.01, n = 75 and r = -0.33, p < 0.01, n = 75, respectively). Out of pinene enantiomers, only (1S)-(-)-α-pinene showed some negative correlation with monthly temperature (r = -0.26, p < 0.05, n = 75). Different patterns of essential oil and pinene dynamics during growing season within separate habitats suggested that some genetic variables of Picea abies might be involved.

Response of juveniles of seven forest tree species and their populations to different combinations of simulated climate change-related stressors: spring-frost, heat, drought, increased UV radiation and ozone concentration under elevated CO2 level.

The study aimed to assess response of juvenile progeny of seven forest tree species, Pinus sylvestris, Picea abies, Betula pendula, Alnus glutinosa, Populus tremula, Quercus robur and Fraxinus excelsior, and their populations to different combinations of climate change-related multiple stressors, simulated in a phytotron under elevated CO2 concentration: (1) heat + elevated humidity (HW); (2) heat + frost + drought (HFD); (3) heat + elevated humidity + increased UV-B radiation doses + elevated ozone concentration (HWUO); and (4) heat + frost + drought + increased UV-B radiation doses + elevated ozone concentration (HFDUO). Effects of the complex treatments, species and species-by-treatment interaction were highly significant in most of the growth, physiological and biochemical traits studied, indicating general and species-specific responses to the applied treatments. For deciduous trees, height increment was much higher under HW treatment than in ambient conditions (control) indicating a positive effect of elevated temperature and better water and CO2 availability. HFD treatment caused reduction of height increment in comparison to HW treatment in most species except for Q. robur and F. excelsior which benefited from lower humidity. Treatments HWUO and HFDUO have caused substantial damages to leaves in fast growing deciduous P. tremula, A. glutinosa and B. pendula, and resulted in their lower height increment than in HW treatment, although it was the same or even higher than that in the control. Rates of photosynthesis in most of the tree species were greatest in HFD treatment. A lower photosynthetic rate (compared to control) was observed in B. pendula, P. tremula and F. excelsior in HW treatment, and in most species-in HWUO treatment. Compared to control, intrinsic water use efficiency in all treatments was significantly lower in P. tremula, A. glutinosa and F. excelsior and higher in conifers P. sylvestris and P. abies. Significant population-by-treatment interactions found for most traits showed variation in response of populations, implying that this reflects adaptive potential of each tree species. The observed responses may not always be considered as adaptive as deteriorating growth of some populations or species may lead to loss of their competitiveness thus compromising regeneration and natural successions.

Between species diversity of Hypericum perforatum and H. maculatum by the content of bioactive compounds.

The objective of present study was to establish and compare the contents of secondary metabolites of two Hypericum species, H. perforatum and H. maculatum, native to Lithuania, and to evaluate factors predetermining their variation with some practical implications for utilization and conservation. The HPLC analysis of the ethanolic extracts of the studied species showed some regularity in their composition. Both species contained chlorogenic acid, hyperoside, quercitrin, quercetin and hypericin. The presence of rutin and hyperforin was observed only in H. perforatum. The quantitative analysis showed higher content of quercitrin in H. perforatum, than in H. maculatum, whereas the differences in the contents of quercetin, hypericin and chlorogenic acid were not statistically significant between the species. H. maculatum contained a significantly higher content of hyperoside than H. perforatum. The data on phytochemical analysis suggest almost equivalent use of both H. perforatum and H. maculatum extracts in the food industry, cosmetics and pharmaceutics.

[Research of the amounts of flavonoids accumulated in the buds of single-styled hawthorn]. / Flavonoidu kiekio tyrimas vienapiestes gudobeles pumpuruose.

OBJECTIVE: To determine the amounts of flavonoids accumulated in the buds of single-styled hawthorn (Crataegus monogyna Jacq.); and to establish the possibility of usage of hawthorn buds in pharmacy practice. MATERIAL AND METHODS: Different examples of hawthorn buds collected in 2001, 2002 and 2003 from some Lithuanian regions: Akmene (V1, V2, V3), Kedainiai (S1, S2, S3), Klaipeda (M1), Lazdijai (D1, D2, D3) and Vilnius (VL1, VL2) from branches of lower storey. Examples were extracted with ethanol and flavonoids were measured by spectroscopic method and by high performance liquid chromatography. RESULTS: From 0.72 to 1.89% of flavonoids are found in the hawthorn buds. The dominating flavonoid is vitexin-2-O-rhamnosid (6.72-10.91 milligrams in one gram of dried crude drug). Other flavonoids are: vitexin: 0.88-6.53 milligrams/gram, hyperosid: 0.85-2.70 milligrams/gram, rutin: 0.72-2.10 milligrams/gram and quercitrin: 0.82-1.01 milligrams/gram. CONCLUSION: The marks of phytochemical compounds of Lithuanian single-styled hawthorn buds corroborated the theory of possibility to use hawthorn buds as pharmaceutical crude drug and form sufficient ground for planning phytochemical and pharmacological researches of new hawthorn crude drug.

[Research of the amounts of flavonoids accumulated in the crude drug of single-styled hawthorn]. / Flavonoidu kiekio tyrimas vienapiestes gudobeles vaistineje augalineje zaliavoje.

UNLABELLED: Leaves, flowers and fruits of single-styled hawthorn are used for production of pharmaceuticals with cardiotonic, coronary vasodilatoric and hypotensive action. Amount of flavonoids accumulated in crude drug of hawthorn is a parameter of quality. OBJECTIVE: To establish the difference in amounts of flavonoids accumulated in the crude drug of single-styled hawthorn. MATERIALS AND METHODS: Different examples of crude material from some Lithuanian regions were extracted with ethanol; flavonoids were measured by spectroscopic method and by HPLC (high performance liquid chromatography). RESULTS: From 0.77% to 2.80% of flavonoids are determined in the leaves and flowers; amounts of hyperosid were 0.16-0.87%; vitexin-o-rhamnosid 0.14-0.40%; rutin 0.10-0.19%. Flavonoids maximum percent was determinate in crude material examples collected at the single-styled hawthorn flowering time; the decline of amounts of flavonoids accumulated in the crude material of hawthorn was determined from May to August (realization of correlation coefficient r=-0.90). Hyperosid maximum percent was determined in crude material examples collected at flowering time; the decline of amounts of hyperosid accumulated in the crude material was determined from May to August (r=-0,94). Amounts of rutin and vitexin-o-rhamnosid in leaves were less than flowers; there was the decline of amounts of rutin and vitexin-o-rhamnosid in leaves from May to August. The percent of flavonoids 0.09 to 0.24 is determined in fruits. CONCLUSION: Lithuanian single-styled hawthorn populations can supply quality crude material from May to August; the bigger amounts of flavonoids were determined in crude drug examples collected at flowering time.

[Phytochemical research of hawthorn species introduced in Lithuania]. / Lietuvoje introdukuotu gudobeliu rusiu ziedu ir lapu fitocheminis tyrimas.

UNLABELLED: The natural vegetable resources of Republic of Lithuania can be supplied by imported hawthorn species, subspecies and varieties, not growing naturally in the territory of country, but well growing and able to produce the crude drug with local conditions of the climate. OBJECTIVE: Determinate amounts of phenolic substances (favonoid glycosides and phenolic acids) in the flowers and leaves of collected species of genus Crataegus and Crataegomespilus. MATERIALS AND METHODS: Different examples of crude drug from some Lithuanian regions were extracted with 96% of ethanol and flavonoids were detected spectroscopically and with HPLC (High performance liquid chromatography). RESULTS: In leaves and flowers were found different quantities of common amounts of flavonoids (1.07-2.53%) and different quantities of individual substances: flavonoids (hyperosid, vitexin-O-rhamnosid, rutin, quercitrin, quercetin, vitexin) and phenolic acids (chlorogenic and caffeic acids). CONCLUSION: Flowers and leaves of hawthorn species, introduced in Lithuania, can be used for preparation of crude drugs procurements; the marks of phytochemical compounds of phenotypes from different genetic lines are sufficient grounds for planning of production of crude drug with special phytotherapeutic influences.