VARIATION AND LOCALIZATION OF FLAVONOID AGLYCONES IN HELIANTHUS ANNUUS (COMPOSITAE)

Flavonoid aglycone variation within Helianthus annuus, a species widely distributed throughout North America, was analyzed. Flavonoid aglycones of H. annuus consist of two types, flavones and chalcones. The flavone aglycones are sequestered in glandular trichomes that occur on both leaf surfaces, whereas the chalcone aglycones appear to be incorporated in the waxy leaf cuticle. Considerable variation in flavonoid profile was observed with some plants exhibiting as few as one, and others as many as seven of the eight aglycones detected. No definable phytogeographic patterns were observed for this flavonoid variation. Flavonoid aglycone variation also did not differentiate the infraspecific taxa within H. annuus.     

POPULATION STUDIES AMONG SONORAN DESERT SPECIES OF PARMELIA SUBG. XANTHOPARMELIA (PARMELIACEAE)

Species of Parmelia subg. Xanthoparmelia exhibit partial morphological convergence in a region of sympatry in the northern portion of the Sonoran Desert. Within the isidiate morph of the population, chemotypic variation is present. These chemotypes exhibit habitat selection among different rock substrates within a region of relatively uniform climate and topography. Partial habitat selection is also demonstrable for the chemotypes of the nonisidiate morph in the population. Other morphological characters are useful in segregating portions of the isidiate and nonisidiate morphs into chemically distinct groups that have previously been assumed to represent only chemotypic variation.     

Environmental and genetic components of flavonoid variation in red oak,Quercus rubra

Two elevation-related chemotypes of red oak (Quercus rubra L.) occur in the southern Appalachians. Studies were carried out to evaluate the effects of genetic and environmental factors on these flavonoid profiles. Low elevation trees which contain only kaempferol and quercetin glycosides were used for assessment of within-tree variation. Within a single tree, sun, shade and branch position have only minor quantitative effects on the flavonoid array. To assess the effect of growing locality on flavonoid profiles we collected red oak acorns from eight locations spanning 1455 m elevation in NC and VA, and divided them equally between nursery plots at 75 and 1140 m in elevation. After five years, leaves of each tree were collected and analysed for flavonoid content and quantity. Myricetin-rich high elevation and quercetin-rich low elevation individuals retain their distinctive profiles regardless of planting size. Some quantitative intrapopulational variation in flavonoid constituents exists between high and low elevation grown trees, but is slight and primarily restricted to minor components. Populations from middle elevations are more heterogeneous than those from either high or low elevations. Some of these trees have lower quantities of myricetin glycosides than high elevation trees while others lack them entirely.     

Chemotype of <Emphasis Type="Italic">Senecio jacobaea</Emphasis> affects damage by pathogens and insect herbivores in the field

The evolution of the diversity of plant secondary metabolites is still poorly understood. To determine whether natural enemies could exert selection on plant secondary chemistry, pathogen infestation and invertebrate herbivory were measured on 10 genotypes of Senecio jacobaea (Tansy Ragwort) at two experimental field sites during a 2-year period. The genotypes represented two chemotypes based on the presence of the pyrrolizidine alkaloids (PA) jacobine and erucifoline. At one site, Heteren, mainly generalist herbivores were present. Here, damage was limited and did not differ among genotypes or chemotypes. At the other site, Meijendel, several specialists attacked the plants. Damage increased during the year, with a peak in July when most damage was caused by the specialist moth Tyria jacobaeae. At this peak there was no difference in damage among chemotypes. In the months prior to T. jacobaeae damage, chemotypes with jacobine were more severely attacked by specialists than the chemotypes without jacobine. Total damage during that period was positively correlated with both total PA concentration and jacobine concentration. Probably plant vigor also played a role in host preference since damage per individual plant was positively correlated with plant size. Our results suggest that total PA concentration and specifically jacobine had a positive effect on specialist feeding, indicating ecological costs involved in the production of PAs. Ecological costs related to plant secondary compounds could explain why not all individuals produce high levels of these compounds. In addition, differences in specialist herbivore pressures among sites may contribute to the variation in secondary metabolites among populations.     

High chemical diversity of a plant species is accompanied by increased chemical defence in invasive populations

Mechanisms contributing to the invasive success of plants are still only partly understood. A main assumption is that an escape from specialized enemies in introduced ranges allows a reduction of chemical defences resulting in an increase in growth and reproduction and thus increased competitive ability of introduced plants. Not only variation in concentration but also variation in composition of chemical compounds between individuals may be a key advantage for plants introduced to novel areas impeding adaptation of different plant antagonists. To investigate quantitative and qualitative variation of putative defence compounds and investment of resources in growth, we conducted a common garden experiment in the native range with seeds of 13 native and 9 introduced populations of Tanacetum vulgare, an aromatic plant forming different chemotypes. After 3.5 months, plants of introduced populations had similar biomass but more stems and higher concentrations of volatile secondary compounds (mainly terpenes) than plants of native populations. Both native and invasive T. vulgare populations exhibited high chemotypic variation with nine chemotypes occurring on both continents, whereas several were found exclusively either in plants originating from the native (n = 10) or invasive (n = 2) range. Due to the known negative effects of many mono- and sesquiterpenes on various organisms, we propose that high concentrations of these secondary compounds accompanied by high chemotypic diversity may facilitate the invasion success of a plant species.     

Proximate causes of altitudinal differences in body size in an agamid lizard

Body size is directly linked to key life history traits such as growth, fecundity, and survivorship. Identifying the causes of body size variation is a critical task in ecological and evolutionary research. Body size variation along altitudinal gradients has received considerable attention; however, the underlying mechanisms are poorly understood. Here, we compared the growth rate and age structure of toad‐headed lizards (Phrynocephalus vlangalii) from two populations found at different elevations in the Qinghai‐Tibetan Plateau. We used mark‐recapture and skeletochronological analysis to identify the potential proximate causes of altitudinal variation in body size. Lizards from the high‐elevation site had higher growth rates and attained slightly larger adult body sizes than lizards from the low‐elevation site. However, newborns produced by high‐elevation females were smaller than those by low‐elevation females. Von Bertalanffy growth estimates predicted high‐elevation individuals would reach sexual maturity at an earlier age and have a lower mean age than low‐elevation individuals. Relatively lower mean age for the high‐elevation population was confirmed using the skeletochronological analysis. These results support the prediction that a larger adult body size of high‐elevation P. vlangalii results from higher growth rates, associated with higher resource availability.     

Chemical Diversity among the Essential Oils of Wild Populations of Stachys lavandulifolia Vahl (Lamiaceae) from Iran

The variation of the essential‐oil composition among ten wild populations of Stachys lavandulifolia Vahl (Lamiaceae), collected from different geographical regions of Iran, was assessed by GC‐FID and GC/MS analyses, and their intraspecific chemical variability was determined. Altogether, 49 compounds were identified in the oils, and a relatively high variation in their contents was found. The major compounds of the essential oils were myrcene (0.0–26.2%), limonene (0.0–24.5%), germacrene D (4.2–19.3%), bicyclogermacrene (1.6–18.0%), δ‐cadinene (6.5–16.0%), pulegone (0.0–15.1%), (Z)‐hex‐3‐enyl tiglate (0.0–15.1%), (E)‐caryophyllene (0.0–12.9), α‐zingiberene (0.2–12.2%), and spathulenol (1.6–11.1%). For the determination of the chemotypes and the chemical variability, the essential‐oil components were subjected to cluster analysis (CA). The five different chemotypes characterized were Chemotype I (germacrene D/bicyclogermacrene), Chemotype II (germacrene D/spathulenol), Chemotype III (limonene/δ‐cadinene), Chemotype IV (pulegone), and Chemotype V (α‐zingiberene). The high chemical variation among the populations according to their geographical and bioclimatic distribution imposes that conservation strategies of populations should be made appropriately, taking into account these factors. The in situ and ex situ conservation strategies should concern all populations representing the different chemotypes.     

Phenotypic Variations in the Foliar Chemical Profile of Persea americana Mill. cv. Hass

The Hass avocado tree Persea americana cv. Hass was derived from a single hybrid tree of P. americana var. drymifolia and P. americana var. guatemalensis, and it is propagated clonally by grafting. This cultivar is the most widely planted in the world but its profile of secondary metabolites has been studied rarely despite of its importance in plant protection. We illustrate the variability of the volatilome of mature leaves by describing the average chemical composition and the phenotypic variability found in 70 trees. Contrary to the uniformity expected in the Hass cultivar, high variability coefficients were found for most of the 36 detected foliar volatile compounds; furthermore we found six chemotypes grouping the foliar phenotypes of the sampled trees using hierarchical cluster analysis. About 48% of trees were grouped in one chemotype; five chemotypes grouped the remaining trees. The compounds that determined these chemotypes were: estragole, α‐farnesene, β‐caryophyllene, germacrene D, α‐cubebene and eugenol. This striking variation in a cultivar propagated clonally is discussed in terms of somatic mutation.     

Speciation in lichens of the Ramalin a siliquosa complex (Ascomycotina,Ramalinaceae): gene flow and reproductive isolation

The lichens of the rock-inhabiting Ramalina siliquosa complex of maritime cliffs of western Europe are differentiated into six chemotypes characterized by a replacement series of β-orcinol depsidones. These chemotypes are sharply zoned from the most sheltered to the most exposed positions in the supralittoral zone. Since artificial crosses in lichens are impossible, the breeding system was investigated by an analysis of the progeny of matemals from nature—some growing in pure zones and others growing intermixed with different chemotypes where zones overlap. Overall, the chemotypes are highly reproductively isolated, the progeny tending to be chemically identical to their respective maternal individuals. Only one pair of chemotypes appears to represent a clear polymorphism; the remainder are apparently ecologically differentiated sibling species. Two chemotypes, which interbreed where their zones overlap, are apparently kept distinct by strong selection against unadapted sporelings in the “wrong” niche. Spermatia function over only short distances, also contributing to the reproductive isolation.     

Essential oils from New Zealand manuka: triketone and other chemotypes of Leptospermum scoparium

The triketone chemotype of manuka, Leptospermum scoparium (Myrtaceae), is commercially important because of its antimicrobial activity. Oils from 36 individual plants on the East Cape of New Zealand all showed similar high triketone contents (>20% total triketones) with little seasonal variation. Analyses of oils from 261 individual manuka plants collected from 87 sites throughout New Zealand showed that the high triketone chemotype was localised on the East Cape, although oils with triketone levels up to 20% were found in the Marlborough Sounds area of the South Island. Cluster analysis revealed other chemotypes localised on other areas. Ten further chemotypes are described: alpha-pinene; sesquiterpene-rich with high myrcene; sesquiterpene-rich with elevated caryophyllene and humulene; sesquiterpene-rich with an unidentified sesquiterpene hydrocarbon; high geranyl acetate; sesquiterpene-rich with high gamma-ylangene + alpha-copaene and elevated triketones; sesquiterpene-rich with no distinctive components; sesquiterpene-rich with high trans-methyl cinnamate; high linalol; and sesquiterpene-rich with elevated elemene and selinene. Some of the chemotypes contained aroma compounds at relatively high levels, with a geranyl acetate-rich oil being most notable. Possible origins for this complex array of chemotypes are proposed.     

Differential effects of bacterial lipopolysaccharides upon neutrophil function

Lipopolysaccharide (LPS) is a potent inflammatory agent which augments neutrophil sensitivity to subsequent inflammatory stimuli. In this study, the effects of structurally different LPS types upon neutrophil effector functions were examined. Rough LPS types, which have lost the O-polysaccharide moiety, were found to act more rapidly than smooth LPS types in stimulating neutrophil β₂ integrin activity and fMLP-induced respiratory burst. These findings suggest an involvement of the O-polysaccharide region of LPS in regulating neutrophil responsiveness to different LPS chemotypes with important implications for the mechanisms underlying regulation of the inflammatory response in conditions associated with elevation of LPS in plasma, e.g. septic shock or acute respiratory distress syndrome.     

Problems in chemotaxonomy: The influence of varying soil conditions,of geographical and individual variants upon the distribution of certain substances in chromatographed extracts ofmonarda fistulosa

Summary The effect of growing a clone ofMoranda fistulosa, in different soil conditions upon their chromatographic patterns is tested. Several plants of a single clone were grown in soils of different compositions. Extracts of leaves were chromatographed for aglycones and glycosides after two and seven months of growth. The resulting chromatographs showed that there was no significant variation of distinguishing compounds.Chromatograms of geographical and individual variants however resulted in a considerable variation in their patterns. These data show, that there is not a close coincidence between them and the taxonomic conclusions based upon morphology. The use of chromatographic data in the genusMonarda is probably justified only once the totality of variation within this taxon has been established.     

FLAVONOID DIVERSIFICATION IN THE ACHILLEA AGERATIFOLIA AND A. CLAVENNAE GROUPS (ASTERACEAE)

The Achillea ageratifolia and A. clavennae groups (Asteraceae, tribe Anthemideae) were investigated for intra- and interspecific leaf flavonoid variation. Numerous flavonoid glycosides, mostly unidentified, and variously methylated free aglycones based on scutellarein, 6-hydroxyluteolin, quercetagetin, and 6-hydroxy-kaempferol were detected. The distribution of methylated aglycones among individuals of A. ageratifolia was used to model the putative biosynthetic pathways. Several steps in the biosynthesis probably involve enzymes (methyltransferases) with strict positional specificities and broader substrate specificities. Geographical patterns in the flavonoid content of A. ageratifolia were only partly consistent with the current concept of A. ageratifolia. Different flavonoid profiles in A. ageratifolia do not appear to be of crucial importance in its adaptation to different habitats. The species of the A. clavennae group were clearly separable on flavonoid profiles and the presence of a hybrid population was supported by the flavonoid data. Interspecific differences in number of glycosides in relation to the number of free aglycones are discussed.     

Variation in Gender and Flower‐Size Dimorphism in the Dioecious Tree Dombeya ciliata,an Endemic to La Reunion Island1

The genus Dombeya is an important component of tropical forests in west Africa, Madagascar, and the Mascarene islands, but little is known about the reproductive biology of its members. In this study, we quantified gender variation and flower‐size variation in three populations of the dioecious tree Dombeya ciliata, an endemic of La Reunion Island (Indian Ocean). Variation among the three populations was observed for three characters. First, the high elevation population had strict males and females, whereas in the two low elevation populations, males were inconstant, with > 50 percent of the males set a small number of fruit. Second, females showed greater seed production at high elevation. Third, flower‐size dimorphism was greater at high elevation because male flowers were larger than female flowers.     

Intraspecific Variability of the Essential Oil of Ziziphora clinopodioides ssp. rigida from Iran

Hydrodistillated essential oils of Ziziphora clinopodioides ssp. rigida from nine populations of the Lashgardar protected region (Hamedan Province, Iran) were analyzed by using GC and GC/MS techniques to determine the intraspecific chemical variability. Altogether, 39 compounds were identified in the oils, and a relatively high variation in their contents was found. The main constituents of the essential oils were pulegone (0.7–44.5%), 1,8‐cineole (2.1–26.0%), neomenthol (2.5–22.5%), 4‐terpineol (0.0–9.9%), 1‐terpineol (0.0–13.2%), neomenthyl acetate (0.0–7.1%), and piperitenone (0.0–5.4%). For the determination of the chemotypes and the intraspecific chemical variability, the essential oil components were subjected to cluster analysis (CA). The five different chemotypes characterized were Chemotype I (pulegone/neomenthol), Chemotype II (pulegone), Chemotype III (pulegone/1,8‐cineole), Chemotype IV (neomenthol), and Chemotype V (1,8‐cineole/4‐terpineol).     

Use of intraspecific variation in thermal responses for estimating an elevational cline in the timing of cold hardening in a sub‐boreal conifer

To avoid winter frost damage, evergreen coniferous species develop cold hardiness with suitable phenology for the local climate regime. Along the elevational gradient, a genetic cline in autumn phenology is often recognised among coniferous populations, but further quantification of evolutionary adaptation related to the local environment and its responsible signals generating the phenological variation are poorly understood. We evaluated the timing of cold hardening among populations of Abies sachalinensis, based on time series freezing tests using trees derived from four seed source populations × three planting sites. Furthermore, we constructed a model to estimate the development of hardening from field temperatures and the intraspecific variations occurring during this process. An elevational cline was detected such that high‐elevation populations developed cold hardiness earlier than low‐elevation populations, representing significant genetic control. Because development occurred earlier at high‐elevation planting sites, the genetic trend across elevation overlapped with the environmental trend. Based on the trade‐off between later hardening to lengthen the active growth period and earlier hardening to avoid frost damage, this genetic cline would be adaptive to the local climate. Our modelling approach estimated intraspecific variation in two model components: the threshold temperature, which was the criterion for determining whether the trees accumulated the thermal value, and the chilling requirement for trees to achieve adequate cold hardiness. A higher threshold temperature and a lower chilling requirement could be responsible for the earlier phenology of the high‐elevation population. These thermal responses may be one of the important factors driving the elevation‐dependent adaptation of A. sachalinensis.     

Genetic variation in flowering phenology and reproductive performance in a Mediterranean high‐mountain specialist,Armeria caespitosa (Plumbaginaceae)

Adaptive responses to past climate change may play an important role in the persistence of high‐mountain plants, which are vulnerable to global warming. Armeria caespitosa is a high‐mountain plant, endemic to the Iberian Central Range. Differences in abiotic environment along the elevational gradient impose two opposing stress gradients (i.e. water stress and duration of the growth season) on the species. Furthermore, the species is found in two interspersed, contrasting microhabitats (rocky outcrops and dry cryophilic grasslands) that have different effects on plants depending of the elevation. As a result of this, the species shows great among‐population variation in many reproductive and vegetative traits. We used a common garden approach to determine whether this phenotypic variation has a genetic basis or is the result of plastic responses shaped by heterogeneous environmental conditions. Plants from the high‐elevation edge and dry cryophilic grasslands flowered earlier and produced more viable fruits but were smaller. These results confirm that among‐population variation in flowering phenology and reproductive performance traits in A. caespitosa is partially genetically based. The results also show that the stronger selection response in favour of early‐flowering individuals in populations at the low‐elevation edge did not correspond with the greater proportion of early‐flowering individuals. Genetic variability associated with flowering onset may be relevant in coping with the impacts of ongoing global warming. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 176 , 384–395.     

Do co-occurring plant species adapt to one another? The response of <Emphasis Type="Italic">Bromus erectus</Emphasis> to the presence of different <Emphasis Type="Italic">Thymus vulgaris</Emphasis> chemotypes

Local modification of the soil environment by individual plants may affect the performance and composition of associated plant species. The aromatic plant Thymus vulgaris has the potential to modify the soil through leaching of water-soluble compounds from leaves and litter decomposition. In southern France, six different thyme chemotypes can be distinguished based on the dominant monoterpene in the essential oil, which is either phenolic or non-phenolic in structure. We examine how soils from within and away from thyme patches in sites dominated by either phenolic or non-phenolic chemotypes affect germination, growth and reproduction of the associated grass species Bromus erectus. To do so, we collected seeds of B. erectus from three phenolic and three non-phenolic sites. Seeds and seedlings were grown on soils from these sites in a reciprocal transplant type experiment in the glasshouse. Brome of non-phenolic origin performed significantly better on its home soil than on soil from a different non-phenolic or a phenolic site. This response to local chemotypes was only observed on soil collected directly underneath thyme plants and not on soil in the same site (<5 m away) but where no thyme plants were present. This is preliminary evidence that brome plants show an adaptive response to soil modifications mediated by the local thyme chemotypes. Reproductive effort was consistently higher in brome of phenolic origin than in brome of non-phenolic origin (on both thyme- and grass-soil), indicating that life-history variation may be related to environmental factors which also contribute to the spatial differentiation of thyme chemotypes. Moreover, we found that brome growing on thyme-soil in general was heavier than when growing on grass-soil, regardless of the origin of the brome plants. This is concordant with thyme-soil containing higher amounts of organic matter and nitrogen than grass-soil. Our results indicate that patterns of genetic differentiation and local adaptation may modify competitive interactions and possible facilitation effects in natural communities.     

Mentha suaveolens Ehrh. Chemotypes in Eastern Iberian Peninsula: Essential Oil Variation and Relation with Ecological Factors

Essential oil (EO) extracts coming from two representative populations of Mentha suaveolens Ehrh . subesp. suaveolens in Eastern Iberian Peninsula were analyzed by gas chromatography coupled with mass spectrometry and flame ion detector. Plant sampling was carried out in the morning and evening in order to study diurnal variation in EO profiles. Likewise, leaves and inflorescences were analyzed separately. Two chemotypes corresponding to each one of the populations were identified, with piperitenone oxide (35.2 – 74.3%) and piperitone oxide (83.9 – 91.3%), respectively, as major compounds. Once different chemotypes were identified, canonical correspondence analysis was employed to evaluate the effect of the bioclimatic and edaphic factors recorded in each location on the observed differences. Statistical analysis suggested that these chemotypes were closely related to specific environmental factors, mainly the bioclimatic ones. Concretely, piperitenone oxide chemotype can be associated to supramediterranean bioclimatic conditions and soils with major salinity and water field capacity. On the other hand, the most volatile fraction (hydrocarbon monoterpenes) reached its higher level in the morning; specifically, a noticeable amount of limonene was found in morning samples of flowers (4.8 – 10.6%). This fact can be related to ecological role of volatile compounds in order to attract pollinator insects.