Essential oil composition and preliminary molecular study of four Hungarian Thymus species

Chemical and genetic differences of twenty taxa belonging to four Thymus species were studied in order to determine whether molecular characters and essential oil components could be used as taxonomic markers and to examine the correlation between them. Plant samples, representing different taxa and geographic regions, were collected from experimentally grown populations. Essential oil samples were analysed by GC/MS and cluster analysis of volatile composition resulted in segregation of thymol chemotypes from sesquiterpenic ones. Thymol was characteristic for all the populations of Thymus glabrescens and T. pannonicus as well as for certain taxa belonging to T. praecox and T. pulegioides. Sesquiterpenes occurred in only two taxa of T. glabrescens, in each sample of T. praecox and in three taxa of T. pulegioides. Plant samples were analysed by random amplified polymorphic DNA (RAPD). The obtained dendrogram revealed high gene diversity. The 13 primers resulted 114 polymorphic RAPD bands, and the average percentage of polymorphism was 80.8%. The RAPD dendogram showed separation neither at interspecific nor at interpopulational levels. Therefore, further specific molecular studies involving more taxa are suggested. Partial correlation have been found between molecular and chemical assessments.     

The alpha-terpenyl acetate chemotype of essential oil of Thymus pulegioides L

The composition of the essential oils of Thymus pulegioides L. plants growing wild in Vilnius district (Lithuania) with "ester", lemon and carvacrol (phenol) odours were collected in one location and investigated by GC, GC/MS and IR-spectroscopy. The essential oils with lemon and carvacrol odour belong to corresponding known citral-geraniol and carvacrol chemotypes. The essential oil of plants with "ester" odour containing 50-70% of alpha-terpenyl acetate belong to alpha-terpenyl acetate chemotype, which was not noticed earlier in Thymus pulegioides L. species.     

Intra-population terpene polymorphism of Thymus pulegioides L.: Evidence for seven chemotypes in a German limestone grassland

For Thymus pulegioides L. (Lamiaceae), occurring in almost entire Europe, about 20 different essential oil chemotypes are described in approximately 25 studies. However, only few studies mention chemotype diversity on population level, describing up to five chemotypes growing together. The aim of the study was to investigate the chemotype diversity within one T. pulegioides population of a limestone grassland in Germany. Essential oil compounds from samples of 93 thyme cushions were extracted with solid phase extraction (SPE) and analysed by GC-MS. Cluster analysis and nonmetric multi-dimensional scaling (NMDS) of the samples’ essential oil composition revealed the existence of seven different chemotypes: linalyl acetate-chemotype (56 individuals), geraniol-citral-linalyl acetate-chemotype (5), geraniol-citral-chemotype (7), thymol-chemotype (8), carvacrol-chemotype (2), linalool-chemotype (1), and β-caryophyllene-germacrene D-β-bisabolene-chemotype (14). The pattern of major monoterpenes from all chemotypes could be explained by the inhibition of specific steps in monoterpene biosynthesis. The sesquiterpene-dominant chemotype might be caused by a block in a very early step of monoterpene production or a regulatory/channeling mechanism. While the geraniol-citral-linalyl acetate-chemotype has been found for the first time, the other chemotypes have been found in other regions and the β-caryophyllene-germacrene D-β-bisabolene-type is similar to other sesquiterpene chemotypes rarely found in other areas. The large intra-population chemical polymorphism in this study raises questions about the chemotype diversity of thyme in other regions of Germany and potential correlations between essential oil composition and abiotic factors or biotic interactions.     

Earlier spring staging in Iceland amongst Greenland White-fronted Geese <Emphasis Type="Italic">Anser albifrons flavirostris</Emphasis> achieved without cost to refuelling rates

Greenland White-fronted Geese wintering in Ireland and Britain stage for 3 weeks in Iceland in spring before migrating onwards to breeding areas in west Greenland. The geese now depart their wintering quarters 12–15 days earlier than in 1973 because they attain necessary fat stores earlier than in previous years. Icelandic temperatures at critical midway staging areas have shown no significant change since 1973, creating a potential mismatch in food availability along the migratory route. Greenland White-fronted Geese have shifted from consuming below-ground plant storage organs in Iceland in spring to grazing managed hayfields created since the 1950s where fresh grass shoot growth occurs despite sub-zero temperatures, when traditional natural foods are physically inaccessible to staging geese due to frozen substrates. Rates of fat accumulation (measured by field scores of abdominal profiles) and mass change (measured in captured geese) were the same in the springs of 1997, 1998 and 1999 as in that of 2007 when the migration episode was 10 days earlier. Hence, earlier arrival in Iceland in 2007 did not occur at cost to refuelling rates there. The shift to acquiring energy from artificial grasslands has enabled Greenland White-fronted Geese to arrive in Iceland earlier, but has apparently not impaired their ability to accumulate fat reserves required for onwards migration, which occurs at the same rate, only earlier in the spring.     

Distribution,abundance and biology of polar cod, <Emphasis Type="Italic">Boreogadus saida</Emphasis>, in Iceland–East Greenland waters

Recent warming has caused a northern extension in the distribution of many southern fish species in Icelandic waters. Polar cod (Boreogadus saida) around Iceland are near the southern limit of their distribution, but are poorly studied in the area. Therefore, material sampled during demersal fish surveys in March 1985–2013 and in pelagic 0-group surveys in Iceland–East Greenland waters in August–September 1974–2003 was used to investigate their distribution, abundance and biology. Demersal polar cod were most often caught on the outer shelf to the north-west and north of Iceland, but during years of widest distribution and highest abundance, they were caught farther to the east on the northern shelf. Pelagic 0-group polar cod were only caught sporadically and mainly confined to the waters off the north-west shelf of Iceland and the East Greenland shelf (southern Denmark Strait). In demersal hauls, the number of stations with polar cod decreased with increasing bottom temperature and polar cod were most widely distributed in the years 1989, 1994 and 1995. Highest numbers of demersal polar cod per haul were caught at temperatures of ?0.5 to 2.5 °C and at 200–450 m depth. The length of demersal polar cod ranged from 5 to 32 cm, while the fish caught in the pelagic trawl ranged from 2.2 to 19 cm. The polar cod in the subarctic waters north of Iceland most likely originate from the waters off East Greenland and further warming and decline in sea ice may eventually lead to the disappearance of polar cod from Icelandic waters.     

Genetic variation at enzyme loci in North Atlantic minke whales,Balaenoptera acutorostrata

Electrophoretic variation within and between North Atlantic minke whale samples(Balaenoptera acutorostrata) from West Greenland, Iceland, and Norway was investigated. In the West Greenland samples, 28 enzyme systems were examined, representing 36 loci, of which 6 were found to be polymorphic. In Icelandic and Norwegian samples, 22 enzyme systems were examined, representing 29 loci, of which 6 and 5 were found to be polymorphic, respectively. The average heterozygosity was 0.058 (SE=0.024) in samples from West Greenland, 0.074 (SE=0.028) in samples from Iceland, and 0.054 (SE=0.023) in samples from Norway. No significant deviations from the expected Hardy—Weinberg genotypic frequencies, within samples taken from the same area, were found. Significant differences in allele frequencies were observed, however, between samples from the three different areas. The average Nei's genetic distance was 0.014 and the averageF _st value was 0.126. The genetic differences between the samples from the different areas indicate that those from West Greenland, Iceland, and Norway represented different breeding populations.     

Composition of the founding population of Iceland: biological distance and morphological variation in early historic Atlantic Europe

We examined the composition of the founding population of Iceland through the study of morphological traits in skeletons from Iceland, Ireland, Norway, and Greenland. This is the first study to address this issue from the Settlement Period of Iceland and contemporary samples from Ireland. We pose the following questions: 1) Was the founding population of Iceland of mixed or homogeneous origin? 2) Is there evidence for a significant Irish cohort in the founding population, as suggested in medieval Icelandic literature? Analysis of biodistance revealed that both Settlement Age and later samples from Iceland showed a greater degree of phenetic similarity to contemporary Viking Age Norwegians than to samples obtained from early medieval Ireland. Analysis of among‐individual morphological variation showed that the Settlement Age population of Iceland did not exhibit an increase in variation in comparison to other populations in the sample, suggesting a relatively homogenous origin. However, estimation of admixture between the Irish and Norwegian populations indicated that 66% of the Icelandic settlers were of Norwegian origin. Comparison of the Icelandic samples to hybrid samples produced by resampling the Viking Age Norwegian and early medieval Irish samples revealed that the Icelandic samples are much closer to the Norwegian samples than expected, based on a 66:34 mixture of Norwegian and Irish settlers. We conclude that the Settlement Age population of Iceland was predominantly (60–90%) of Norwegian origin. Although this population was relatively homogenous, our results do not preclude significant contributions from Ireland as well as other sources not represented in our analysis. Am J Phys Anthropol, 2003. © 2003 Wiley‐Liss, Inc.     

Volatile Oil Yield and Composition,Total Phenolic Content,Antioxidant Activity and Secondary Metabolite Content of Collected Thymus praecox Species in Rize

In this study, the volatile oil yield (Clevenger), volatile oil (VO) composition (Gas Chromatography), phenolic contents (UV-VIS Spectrophotometer), antioxidant activities (UV-VIS Spectrophotometer) and secondary metabolite content (High Pressure Liquid Chromatography) of 11 Thymus praecox subspecies were evaluated. The most detected chemical class were oxygenated monoterpenes (55.18–86.1 %) in investigated samples. In the present study rosmarinic acid, isoquercitrin, gallocatechin and thymol could be detected in high amounts. The min. and max. content values of Flora/Field Samples were 1543.241 and 890.3-1425.3 for rosmarinic acid, 139.44-287.894 and 129.9-312.2 for thymol, 38.619-121.424 and 26.3-112.9 for gallocatechin as mg/g DW. Principal Component Analysis was used to differentiate Thymus praecox species regarding volatile oil composition and secondary metabolite content. The results demonstrated that T. praecox collected from the Rize flora and cultivated afterwards showed variability based on investigated characteristics. Finally, the Thymus praecox samples displaying high bioactive compounds present useful information for further investigations and applications.     

Differential resistance to freezing and spatial distribution in a chemically polymorphic plant Thymus vulgaris

Secondary compounds play multiple ecological roles. In this study, we present novel experimental evidence of differential tolerance to freezing temperatures among chemotypes of a chemically polymorphic plant, Thymus vulgaris. Non‐phenolic chemotypes showed a significantly better survival and re‐growth after early‐winter freezing (?10° in early December) than phenolic chemotypes. Comparison of temperature data (1971–2002) at a phenolic and non‐phenolic site showed that whereas early‐winter freezing occurred in 6 years in the non‐phenolic site they never occurred at the phenolic site. Observations of trichome morphology (where the essential oil is stocked) with and without intense freezing indicate that non‐phenolic chemotypes may escape any negative effects of freezing by releasing their essential oil into the atmosphere during severe freezing. The correlation between tolerance of freezing and local temperature regimes strongly suggests that differential freezing resistance is a key ingredient of the distribution of thyme chemotypes in space.     

Seasonal variation in migration strategies used to cross ecological barriers in a nearctic migrant wintering in Africa

Ecological barriers such as oceans, mountain ranges or glaciers can have a substantial influence on the evolution of animal migration. Along the migration flyway connecting breeding sites in the North American Arctic and wintering grounds in Europe or Africa, nearctic species are confronted with significant barriers such as the Atlantic Ocean and the Greenland icecap. Using geolocation devices, we identified wintering areas used by ringed plovers nesting in the Canadian High‐Arctic and investigated migration strategies used by these nearctic migrants along the transatlantic route. The main wintering area of the ringed plovers (n = 20) was located in western Africa. We found contrasting seasonal migration patterns, with ringed plovers minimizing continuous flight distances over the ocean in spring by making a detour to stop in Iceland. In autumn, however, most individuals crossed the ocean in one direct flight from southern Greenland to western Europe, as far as southern Spain. This likely resulted from prevailing anti‐clockwise winds associated with the Icelandic low‐pressure system. Moreover, the plovers we tracked largely circumvented the Greenland icecap in autumn, but in spring, some plovers apparently crossed the icecap above the 65°N. Our study highlighted the importance of Iceland as a stepping‐stone during the spring migration and showed that small nearctic migrants can perform non‐stop transatlantic flights from Greenland to southern Europe.     

Chemical polymorphism of the essential oils from populations of Thymus caespititius grown on the island S. Jorge (Azores)

The compositions of the essential oils obtained from ten populations of Thymus caespititius grown on S. Jorge (Azores) were studied by GC and GC-MS. All the oils analysed were dominated by their monoterpene fraction (69-9%). The sesquiterpene fraction was rather small (4-17%) and consisted mainly of oxygen-containing compounds (3-13%). Enantiomeric ratio of sabinene and alpha-terpineol was investigated in the oils from some populations. (+)-Sabinene was detected with high enatiomeric purity (96 99%). Although ( +)-alpha terpineol was the predominant enantiomer, its purity varied considerably (66-97%). Cluster analysis of all identified components grouped the oils into four main clusters that corresponded with their major components: alpha-terpineol (43-68%), carvacrol (32-52%), thymol (44-58%) and sabinene (41%). The essential oil from Th. caespititius showed a clear chemical polymorphism, that was particularly evident among four populations growing along a 200 m distance at Pico Verde.     

Warming winter effects,fat store accumulation and timing of spring departure of Greenland White-fronted Geese <Emphasis Type="Italic">Anser albifrons flavirostris</Emphasis> from their winter quarters

Migratory geese accumulate energy and nutrient stores in winter to fly to refuelling spring staging areas before onward migration to breeding areas. Mean ground temperatures at two important Greenland White-fronted Geese wintering sites rose in winter and spring by 1.0–1.3°C during 1973–2007. Greenland White-fronted Geese departed the Wexford winter quarters on 3rd April 2007 for Icelandic spring staging areas, the earliest on record, representing a mean advancement of 15 days since 1973, mirrored amongst mean dates of departure amongst Scottish wintering birds that have advanced by 12 days during 1973–2007. Icelandic temperatures at critical midway staging areas en route to Greenland showed no significant change since 1973, suggesting that it is warming on the winter quarters that enable geese to depart earlier, rather than elevated temperatures at ultimate spring staging areas. However, Wexford departure date did not correlate with spring temperature. Data presented here show that Greenland White-fronted Geese have accumulated threshold body stores progressively earlier in spring migration, especially during 1995–2007. Although this did not correlate with ambient temperature, the mean degree of accumulated fat stored by 1st April in each year was a statistically significant predictor of departure date for the wintering population at Wexford. These data support the hypothesis that it is intrinsic factors (i.e. improvements in internal body state resulting from better feeding conditions) that has permitted progressively earlier departure of these geese from Wexford on spring migration, rather than amelioration of spring conditions in Iceland or solely the result of warming of the winter quarters.     

Essential oil polymorphism of Thymus praecox subsp. arcticus on the British Isles

The essential oils of 732 individual plants of Thymus praecox Opiz subsp. arcticus (E. Durand) Jalas (syn. T. drucei Ronn.) collected in Scotland, Ireland, and in the south of England have been analysed by gas chromatography (GC) and mass spectrometry (GC–MS) in order to elucidate the chemical character of this subspecies on the British Isles. In total, 69 components were identified, most of them monoterpenoids and sesquiterpenoids with hedycaryol, linalyl acetate, linalool, the germacradienols, trans-nerolidol, T-cadinol, and β-caryophyllene being the most important compounds. The analysis of the quantitative essential oil data by means of neural networks revealed that T. praecox subsp. arcticus growing in Britain is highly polymorphous. There were 17 chemotypes with the hedycaryol chemotype as the most frequent (24% of the plants), followed by the linalool/linalyl acetate chemotype (22% of the plants) and germacra-1(10),4-dien-6-ol chemotype (18% of the plants). It seems that each part of the British Isles has its special chemotype pattern with 13 chemotypes in Scotland, 11 in Ireland, and 17 in the south of England. An overview of the North Atlantic region of Europe revealed that the polymorphism of T. praecox subsp. arcticus in the essential oil is more distinctive in the southern than in the northern regions, with only 2, 5, and 1 chemotypes in Greenland, Iceland, and Norway, respectively.     

What can morphology and isozymes tell us about the history of the Dryas integrifolia–octopetala complex?

An earlier study of the morphological variation in the circumpolar genus Dryas in Greenland revealed that populations in east Greenland are highly variable and this variation was interpreted as a hybrid zone between D. octopetala and D. integrifolia. It was proposed that populations of Dryas in east Greenland immigrated postglacially from Svalbard. The purpose of our study was to investigate the population structure of the Dryas integrifolia-octopetala complex using a combination of morphological and genetic characters. Populations of Dryas were sampled in Greenland, Svalbard, Iceland and north Scandinavia (Sweden). The morphological traits clearly separated west Greenland from Svalbard, Iceland and north Scandinavia. A large number of individuals from east Greenland and some from north Greenland could be interpreted as hybrids between the morphs. The genetic pattern was, however, partly different as the greatest amount of differentiation among regions was found between west and north Greenland (FRT = 0.65). The highest degree of genetic variation appeared within east Greenland. This was in accordance with the putative hybrid origin of the populations, as concluded based on the morphological results. The total amount of genetic differentiation found among the six regions (FRT = 0.61) was larger than between the two species (FRT = 0.44). The genetic pattern matched the proposed postglacial migration routes.     

Stock structure of Icelandic cod Gadus morhua L. based on otolith chemistry

Otolith chemistry was used to study the stock structure of Icelandic cod Gadus morhua . Otoliths were collected from spawning cod at 12 and 17 different spawning locations around Iceland during the spawning season in 2002 and 2003, respectively. The otolith elemental composition of cod spawning north and south of Iceland differed substantially from each other, as did spawning cod below and above 125 m depth at the main spawning ground in the south of Iceland. Otolith chemistry differed more between locations than among years within a location, indicating temporal stability between the 2 years. The Icelandic cod stock is managed as a single stock. These results suggest, however, that optimal fisheries management may require different management units than are currently present.     

Composition of essential oil of <Emphasis Type="Italic">Artemisia scoparia</Emphasis> Waldst. et Kit. from Buryatia and Mongolia

Essential oil from the epigeal portion of Artemisia scoparia Waldst. et Kit. collected in different regions of Buryatia and Mongolia has been isolated using steam distillation. The yield of essential oil from the stems and leaves of A. scoparia collected in different regions of Buryatia and in the Selenga aymag of Mongolia ranged from 0.06 to 0.40%, that is, the variation was rather high. The composition of the oil was analyzed using chromatography with mass spectrometric detection on a gas chromatography device Agilent Packard HP 6890 N with a quadrupole mass spectrometric detector HP MSD 5973. The total number of compounds detected exceeded 100. The major components of Artemisia scoparia oil were the following: p-cymene (0.6–15.2%), limonene (0.1–6.3%), α-pinene (0.2–10.1%), β-pinene (0.4–8.9%), trans-β-ocimene (0.3–5.4%), caryophyllene (4.6–13.8%), germacrene D (11.5–40.3%), spathulenol (4.0–11.7%), and caryophyllene oxide (4.3–15.6%). Several chemotypes of essential oil from A. scoparia were identified according to the nature of the major components: 1) oil that contained acetylene hydrocarbons (from plants collected in Gilan, Mazandaran, Kashan, and Tigran provinces of Iran, Tajikistan, and the European part of the CIS); 2) oil that contained monoterpenes and aromatic compounds (from plants collected in the Khorasan province of Iran, India, and South Korea); and 3) oil that contained monoterpenes or aromatic and sesquiterpene compounds (from plants collected in Kazakhstan and Mongolia). The major components of A. scoparia essential oil obtained from Buryatian plants belonged to the classes of mono- and sesquiterpene compounds, and thus the oil can be assigned to the third chemotype characteristic of A. scoparia from geographically close regions.     

Introgression and phylogeography of Betula nana (diploid), B. pubescens (tetraploid) and their triploid hybrids in Iceland inferred from cpDNA haplotype variation

Aim The objective was to find direct genetic evidence supporting introgressive hybridization between tetraploid tree birch (Betula pubescens) and diploid dwarf birch (B. nana), via triploid hybrids, and to investigate an association between the introgression and phylogeographical distribution of Icelandic birch. Location Samples were collected from 463 trees in 12 woodlands in Iceland and eight locations in Norway, Sweden, Scotland and Greenland. Methods Ploidy status of individual trees was determined by chromosome counting. Variation in the chloroplast genome was assessed using polymerase chain reaction‐restriction fragment length polymorphism. The geographical distribution of the haplotypes was mapped. The haplotype variation and introgression ratios (IG) were analysed statistically. Results Thirteen haplotypes were identified among Icelandic samples. The most common haplotype (T, 49% occurrence) was present in all ploidy groups and in all woodlands. All common haplotypes were shared between the triploid group and the parental species, indicating introgressive hybridization. This was supported by the statistical analysis of IG indices and the variation components. Considerable differences existed among samples, shaped by isolation by distance and local introgression. An east–west phylogeographical distribution in Iceland was observed. Main conclusions Despite extensive introgression across species and ploidy levels, a biogeographical pattern has been observed, and this may indicate different population histories or multiple origins of Icelandic birch. The chloroplast haplotype diversity found in Iceland resembles that found in birch populations from northern Scandinavia.     

Plant survival in Iceland during periods of glaciation?

Aim The paper addresses the classical question of possible plant survival in Iceland during the last glacial period in the light of a palaeobotanical record from northern Iceland, spanning the period 11,300–9000 BP , including the Younger Dryas stadial. We review the Late Cenozoic fossil plant record, the past debate on glacial plant refugia in Iceland, and the evidence for ice-free areas during the Weichselian. Location The investigated lake sediment record comes from Lake Torfadalsvatn, which is situated in the northwestern part of the Skagi peninsula in northern Iceland. Methods The sediment chronology was constructed from the cccurrence of the Vedde Ash and the Saksunarvatn ash, two well-dated Icelandic tephras, together with the results from five AMS and conventional radiocarbon dates performed on bulk sediment samples. The vegetational reconstruction was based on detailed pollen analysis of the sediment sequence. Results The pollen analysis revealed that many of the taxa present in the area prior to the Younger Dryas stadial continued to produce pollen during that cold event. The more or less immediate reappearance of a few other pollen taxa at the Younger Dryas-Preboreal boundary suggests that these plants also survived, even if they did not produce sufficient pollen to be recorded during the Younger Dryas stadial. Main conclusions We conclude that the relatively high plant diversity found in high Arctic areas and present-day nunataks in Iceland and Greenland, together with the fact that many plant species were able to survive the Younger Dryas stadial on the Skagi peninsula, suggest that species with high tolerance for climate fluctuations also survived the whole Weichselian in Iceland. This conclusion is supported by recent palaeoclimatic data from ice-cores and deep-sea sediments, indicating that Icelandic climate during the last glacial was only occasionally slightly colder than during the Younger Dryas stadial.     

Differentiating Thymus vulgaris chemotypes with ISSR molecular markers

Common thyme (Thymus vulgaris L.) is a Mediterranean evergreen shrub of the Lamiaceae that shows chemical polymorphism in its natural populations. In Mediterranean habitats six basic chemotypes have been described according to the main component in its essential oil: thymol, carvacrol, linalool, geraniol, α-terpineol, thuyanol-4, of which the thymol chemotype is widely used in cultivation and as a spice. In this study ten selected clones were analysed in terms of chemical composition of the volatile oil and genetic fingerprint. The study question was whether individual chemotypes can be distinguished by molecular markers. Chemical composition was analysed by GC-MS, and for assessing the genetic polymorphism ISSR marker system was chosen as it is reliable and easy to use even in case of less studied species. The studied T. vulgaris clones represented five different chemotypes and could be distinguished using 12 ISSR primers. Clones belonging to the thymol chemotype showed the most distinct separation. The study shows the potential of using molecular markers in breeding and selection of T. vulgaris, being able to differentiate different chemotypes.     

银木种内化学类型研究

毛细管气相色谱／质谱／计算机联用仪器分析结果表明,在四川省成都市一人工种植的银木（Ｃｉｎｎａｍｏｍｕｍｓｅｐｔｅｎｔｒｉｏｎａｌｅ）种群中,其枝叶精油主要化学组成在各植株间存在很大差异,发现精油存在１,８－桉叶油素,樟脑,异丁香酚甲醇和９－氧代橙花叔醇等四个化学类型．除异丁香酚甲醚类型外,其余类型均为第一次报道．综观樟属其它种的化学类型研究可见,化学类型在樟属植物中普遍存在种内多型性和种间共性．