Population genetic structure and trait associations in forest savory using molecular,morphological and phytochemical markers

In this investigation, morphological, phytochemical and ISSR markers were used to estimate the relationships among and within seven populations of white savory (Satureja mutica), belonging to four provinces in Iran. The individuals were phenotypically diverse, which stamen length, corolla length, corolla diameter, calyx length, bract length, inflorescence length, calyx length and bracteole width were characteristics with the highest variation. Leaf dimensions were in significant correlation with flower and inflorescence characteristics. Chemical compounds of essential oils were found variable in various individuals and all samples were principally composed of phenolic constituents (carvacrol and/or thymol). As a consequence, the plants were classified into two major chemotypes including carvacrol and thymol. A total of 197 band positions were produced by 14 ISSR primers, of which 176 were found polymorphic with 88.91% polymorphism. ISSR genetic similarity values among individuals ranged between 0.45 and 0.94 which was indicative of a high level of genetic variation. Multiple regression analysis (MRA) revealed that phytochemical compositions as dependent variable, showed statistically significant correlation and in association with leaf and flower traits as independent variable, indicating a main role of leaf and flower on production of these compounds. Also, several ISSR fragments were found associated with some morphological traits and phytochemical compositions. The high diversity within and among populations of S. mutica according to different data systems could provide useful information for conservation and selection of cross-parents in breeding programs.     

Comparative analysis of essential oil compositions in seven populations of Bakhtiarian savory in natural and field conditions

Bakhtiarian savory (Satureja bachtiarica Bunge) is an endemic herb growing wild in Iran with interesting pharmacological and biological properties. In this research, the variability of essential oil content and phytochemical compositions among seven populations of this species was studied in both natural and field conditions. There was a wide interpopulation variability in phytochemical variation of studied populations under both natural and field conditions, indicating the existence of different chemotypes. Six chemical compositions showed high value in studied plants so that carvacrol and thymol were known as major compounds and confirmed by principal component analysis. Ilam population had the highest thymol in both field (77.10 %) and natural (42.32 %) conditions and was known as a thymol chemotype. Beside, Yazd and Shahrekord populations, respectively, showed the highest carvacrol percentages in both field (84.83 and 63.81 %) and natural (72.50 and 58.05 %) conditions and were known as carvacrol chemotypes. In other populations, carvacrol and thymol were the main components and showed partly equal percentage and these populations were known as carvacrol/thymol chemotypes. According to Pearson correlation analysis, carvacrol was negatively correlated with p-cymene, γ-terpinene and thymol (r = ?0.61, ?0.70 and ?0.98, respectively), indicating very high reverse correlation between carvacrol and thymol. In conclusion, the high value of the essential oil compositions in studied S. bachtiarica populations could provide useful information for conservation and selection of cross parents in breeding programs to develop improved cultivars.     

Phytochemical and morphological characterization of Satureja khuzistanica Jamzad populations from Iran

Satureja khuzistanica is an endemic herb growing wild in Iran with interesting pharmacological and biological properties. Here, as an initial step of the domestication process, the variability of phytochemical and morphological traits among 69 individuals of eight natural populations of the plant was studied. The investigated characteristics were the essential oil content and composition, the rosmarinic acid (RA) content, and the leaf and flower morphologies. The Abdanan and Kaver populations showed the highest oil contents. The characterization by GC-FID and GC/MS analyses of the oils revealed that all 69 sampled individuals had carvacrol as the main component with very high contents (89.59-95.41%). The content of RA of the MeOH extracts of S. khuzistanica showed a high level of variability (coefficient of variation (CV) 50.0%) ranging from 0.59% (w/w) in the Paalam population to 1.81% (w/w) in the Abdanan population. The peduncle length and the leaf surface area (CVs of 47.39 and 47.21%, resp.) were the most variable morphological characteristics among the examined populations. The high level of phytochemical and morphological variability among the studied populations suggests a breeding approach during the domestication, to gain new, promising, and homogenous cultivars, attractive for the industry and agriculture.     

Variation of bioactive substances and morphological traits in Hypericum perforatum populations from Northern Turkey

Hypericum perforatum L. has become one of the leading plant-based dietary supplements worldwide and its biological activities have been mainly attributed to hypericin and phenolic contents. The present study was conducted to determine chemical and morphological variability of H. perforatum sampled from different locations of Northern Turkey. The populations were investigated according to the hypericin, chlorogenic acid and the flavonoids, namely rutin, hyperoside, apigenin-7-O-glucoside, quercitrin and quercetin contents and morphological characters including leaf dark gland density, leaf area, leaf length/width ratio and plant height. Significant chemical and morphological variations were detected among the populations. Hypericin content among populations ranged between 0.44 and 2.82 mg/g dry weight, chlorogenic acid between 0.0 and 1.86 mg/g dry weight, rutin between 0.0 and 8.77 mg/g dry weight, hyperoside between 5.41 and 22.28 mg/g dry weight, quercitrin between 1.64 and 3.98 mg/g dry weight and quercetin between 1.01 and 1.76 mg/g dry weight. Hypericin content was found to be correlated positively with leaf dark gland density, however, negatively with leaf area and no correlation was detected between the other morphological traits and bioactive substances examined.     

Heterophylly results in a variety of “spectral signatures” in aquatic plant species

The leaf reflectance spectra (280–887 nm) of two heterophyllous aquatic plant species Polygonum amphibium (L.) and Nuphar luteum (L.) were compared and their relation to physical properties of the leaves examined. In P. amphibium contrasting environmental conditions along water–land gradient affected the majority of anatomical and morphological properties of leaves, but less differences were observed in photosynthetic pigment and total flavonoid contents. Leaf mass per area (LMA), palisade mesophyll, leaf thickness, trichome length and anthocyanin content per dry mass were correlated to the different parts of spectra. In N. luteum natant and submerged leaves differed significantly in all measured parameters. Chlorophyll a, anthocyanin and carotenoid contents per dry mass were related to reflectance in the red region, while leaf thickness, anthocyanin and total flavonoid contents per leaf area were related to reflectance in the near infrared region. Redundancy Analysis (RDA) indicated that in P. amphibium the average length of trichomes and LMA explained 72% and 6% variability of the spectra, whereas in N. luteum anthocyanin content per dry mass, explained 57% variability of the spectra. The comparison of natant leaves of both species showed that they were more similar than different leaf types within the single species.     

Effects of altitude of origin on trichome-mediated anti-herbivore resistance in wild Andean potatoes

The diversity and abundance of herbivores often decreases with increasing altitude. Plants are expected to respond to reduced herbivore pressure by allocating fewer resources to anti-herbivore resistance at higher altitudes. This study uses a greenhouse experiment and parallel bioassay to examine population variability in trichome-mediated defenses across a range of accessions of Solanum berthaultii Hawkes and S. tarijense Hawkes that originated from different altitudes. S. berthaultii bears two types of glandular trichomes, type A and type B, whereas S. tarijense has type A trichomes only, and hairs resembling type B trichomes that are eglandular. Both type A and type B glandular trichomes on S. berthaultii deterred ovipositing female Phthorimaea operculella (Zell.) (Lepidoptera: Gelechiidae). The density of type A, but not type B trichomes decreased with increasing altitude of origin in S. berthaultii populations. The ratio of type A to type B trichomes on the abaxial surface of S. berthaultii negatively affected oviposition and was inversely related to altitude of origin. In S. tarijense, type A trichomes deterred, but eglandular leaf hairs stimulated oviposition. Consequently, the ratio of type A trichomes to eglandular leaf hairs was negatively related to the number of eggs laid. The total numbers of trichomes per leaf generally increased with increasing altitude of origin in S. tarijense whereas the ratio of type A trichomes to eglandular hairs declined. In the oviposition bioassay, we found no direct relationship between resistance and altitude of origin, which may have been due to differences in leaf area at the time of the bioassay. Nevertheless, the results suggest that populations of both plant species that originated from higher altitudes were generally more susceptible to ovipositing P. operculella.     

Floral traits and mating systems in sister species of <Emphasis Type="Italic">Nicotiana</Emphasis>: interpopulational variability and sympatry effects

Mating systems of Angiosperms are important determinants of population genetic structure and evolutionary potential. Nicotiana longiflora and N. plumbaginifolia are self-compatible, sister species, with contrasting floral morphology and can be found in allopatry and sympatry in North Argentina. In two sympatric and 10 nearby allopatric populations we studied their natural interpopulational variability and sympatry effects on corolla length and anther–stigma distance. We also estimated seed set by selfing and via pollinators using pollination treatments. Both corolla length and anther–stigma distance varied significantly among N. longiflora, but not among N. plumbaginifolia populations. We did not detect an effect of sympatry in either species for any of the floral traits studied. Pollination treatments suggest that N. longiflora is mainly an outcrosser, although selfing occurs at some extent. Seed set attributed to pollinators was significantly higher in N. longiflora whereas most seeds in N. plumbaginifolia were sired through self-pollination. In N. plumbaginifolia, selfing seems to assure reproduction in sympatric populations, where floral visitors have a strong preference for N. longiflora. Corolla length was significantly negatively correlated with an increase in the percentage of self-seeds estimated by pollination treatments. Within N. longiflora, anther–stigma distance showed a positive correlation with selfing.     

Il problema della Eredità considerato dal punto di vista chimico-fisico

Abstract

A study on the type and distribution of trichomes, morphological traits of the calyx and content of antioxidant compounds was conducted on Teucrium arduini L. populations in Croatia and in Bosnia and Herzegovina. Non-glandular trichomes, peltate trichomes and small capitate trichomes with four secretory head cells were observed on leaves, stems and calyx, whereas long capitate trichomes with a unicellular head cell were observed exclusively on the calyx. Average calyx length was 5.23–6.51 mm, calyx tube length 2.92–3.68 mm, lower lip length 1.25–2.42 mm and upper lip length 1.89–2.99 mm. The concentration of antioxidant compounds in all populations was the highest in leaves: total polyphenols (TP): 8.43–13.55%; tannins (T): 1.79–7.33%; flavonoids (TF): 0.28–0.40%; phenolic acids (TPA): 2.90–5.07%. Inflorescences generally contained the smallest amount of TP (5.89–11.42%), T (0.87–6.50%) and TPA (1.66–2.27%), while the smallest content of TF was obtained for the majority of stem samples (0.06–0.12%). The researched populations did not differ with regard to the type and overall distribution of the trichomes, though the Bosnian populations have a shorter calyx tube and longer upper lip. Some variability was observed in the TP and T content between sampling sites, while quite high variability was noted in the TF and TPA content.     

浙江省野生蜡梅花部形态变异及其与环境因子的相关性

以浙江省蜡梅自然分布区的5个居群为研究对象,采用巢式方差分析、主成分分析、相关分析、聚类分析等多种分析方法,探究蜡梅花部的形态变异及其与环境因子相关性以及居群间和居群内的表型多样性。结果表明:蜡梅花部性状中除雄蕊长度、雌蕊数、雌蕊长度、内被片数以外,其他12个性状在居群间均有极显著差异。胚珠数的平均变异系数最大,花冠内径的平均变异系数最小,各性状的平均变异系数为15.03%(10.81%—23.38%)。五尖山的表型多样性最丰富,碧东山居群最小,种群间平均表型分化系数为44.38%(1.57%—89.62%),种群内变异大于种群间变异,种群内变异是花部变异的主要来源。主成分分析显示花冠直径、花冠内径、花筒深度和中被片长对种群变异起主要贡献作用。花部多数性状间存在显著或极显著的相关关系,生态因子中花部性状与土壤酸度相关性最大,土壤中花部性状与大量元素相关性最大,微量元素次之,中量元素最小。通过UPGMA聚类分析可以将5个种群分成两组。浙江省野生蜡梅花部形态存在丰富的变异和多样性,花部部分性状与土壤中钙、铁、铜含量和海拔有显著或极显著相关关系。     

Geographic association and temporal variation of chemical and physical defense and leaf damage in Datura stramonium

The evolution of plant defense traits has traditionally been explained trough the “coevolutionary arms race” between plants and herbivores. According to this, specialist herbivores have evolved to cope effectively with the defensive traits of their host plants and may even use them as a cue for host location. We analyzed the geographic association between leaf trichomes, two tropane alkaloids (putative resistance traits), and leaf damage by herbivores in 28 populations of Datura stramonium in central Mexico. Since the specialist leaf beetles Epitrix parvula and Lema trilineata are the main herbivores of D. stramonium in central Mexico, we predicted a positive association between plant defense and leaf damage across populations. Also, if physical environmental conditions (temperature or precipitation) constrain the expression of plant defense, then the geographic variation in leaf damage should be explained partially by the interaction between defensive traits and environmental factors. Furthermore, we studied the temporal and spatial variation in leaf trichome density and leaf damage in five selected populations of D. stramonium sampled in two periods (1997 vs. 2007). We found a positive association between leaf trichomes density and atropine concentration with leaf damage across populations. The interaction between defensive traits and water availability in each locality had a significant effect on the geographic variation in leaf damage. Differences among populations in leaf trichome density are maintained over time. Our results indicate that local plant–herbivore interaction plays an important role in shaping the geographic and temporal variation in plant defense in D. stramonium.     

Micromorphological traits and essential oil of Micromeria longipedunculata Bräuchler (Lamiaceae)

Micromeria longipedunculata Bräuchler (syn. M. parviflora (Vis.) Rchb.) is an endemic Illyric-Balkan plant species from Bosnia and Herzegovina, Montenegro, and Albania. We investigated types and distribution of trichomes, pollen morphology, and chemical composition of essential oil (analysed by GC and GC-MS) in M. longipedunculata. Non-glandular trichomes, peltate trichomes, and two types of capitate trichomes (type 1 composed of one basal epidermal cell, and one head cell with subcuticular space; type 2 composed of one basal epidermal cell, two or three stalk cells, and one head cell with subcuticular space) were observed on leaves, bracteoles, the calyx, corolla, and the stem. The pollen grains had six apertures which were set in the equatorial pollen belt and showed medium reticulate ornamentation. A phytochemical analysis of essential oils from four different localities is characterized by similar chemical composition with spathulenol (23.7–39.5%), piperitone oxide (7.7–12.1%) and piperitone (7.3–8.9%) as the major compounds.     

黄河下游湿地芦苇形态变异研究

 以水盐为主导生态因子,分析了黄河下游湿地芦苇(Phragmites australis)种群形态变异的水平与格局。对基径、株高、叶长、叶宽、节间长、节间数和穗长7个形态特征在15个种群中的变异分析表明：种群内个体间形态差异极显著,7个形态特征在种群内的变异系数从大到小依次为节间长（0.284 6）、叶宽（0.253 6）、穗长（0.244 9）、叶长（0.208 5）、基径（0.187 5）、节间数（0.176 3）、株高（0.165 7）; 7个形态特征在种群间的变异度从大到小依次为株高、叶长、节间长、叶宽、基径、节间数、穗长,但种群间形态的显著差异主要存在于滨州种群（BZH）、淡水种群（ZHG、DPH、NYH,土壤总盐度<0.1%）、盐生种群（HS01、HS02、HS03、HS04、HS05、DWL、KD、GD、LZH、LJ、DLH,土壤总盐度>0.3%）之间,在来自相似生境的种群间形态差异普遍不显著;7个形态特征均与水盐联合影响因子显著相关;以形态特征为基础的聚类分析将研究的15个种群分为显著不同的３类。因此,根据黄河下游湿地芦苇形态变异规律和分化特点,建议将该地区芦苇分为盐生芦苇、淡水芦苇、巨型芦苇3个形态类型。     

The variation and evolution of selected species of solanum section basarthrum

A study of greenhouse-grown and field populations ofSolarium caripense Humboldt & Bonpland ex Dunal,S. tabanoense Correll, andS. trachycarpum Bitter & Sodiro, all diploid species (n = 12) of high altitudes of southern Central America and northern South America, revealed great morphological variation. Polygonographs utilizing seven characters (number of leaflets, leaf index, length of pubescence, number of flowers, anther length, corolla index, and pollen diameter) showed a wide range of variation and led to recognition of six morphologically distinct groups. Hybridizations with greenhouse populations showed that five of the morphological groups are reproductively isolated as well. A complex pattern of genetic variation involving various degrees and combinations of low crossing success, low seed set, lowered F₁ pollen fertility, and nonreciprocal crossability was found. Examinations of meiotic figures from hybrids revealed no gross chromosomal structural differences. Evidence indicates that genetic differences, including gene-cytoplasm interactions, are significant isolating barriers. A key to the species studied plus appropriate taxonomic notes are provided.Solanum heiseri is described as new, andS. trachycarpum is placed inS. sect.Basarthrum seriesCaripensia.     

Inhibitor and substrate activities of sesquiterpene olefins toward (+)-δ-cadinene-8-hydroxylase, a cytochrome P450 monooxygenase (CYP706B1)

Several lines of evidence indicate that (+)-δ-cadinene-8-hydroxylase (CYP706B1) plays an important role in biosynthesis of gossypol in Gossypium arboreum L. ( [Luo et al., 2001] and [Wang et al., 2003]). The catalytically active enzyme has been expressed in yeast microsomes. Some microsomal preparations conjugated the hydroxylated (+)-δ-cadinene to a moiety that has not yet been identified. However, when microsomes were treated with n-octyl-β-d-glucoside (OG), a non-ionic detergent, (+)-δ-cadinene was reproducibly converted to the free alcohol, 8-hydroxy-(+)-δ-cadinene. OG had little effect on K_m and slightly stimulated apparent V_max. Enzymic activity was more than 10-fold more sensitive to inhibition by the N-substituted imidazole clotrimazole than to miconazole. Sesquiterpene olefins (−)-δ-cadinene, (−)-α-cubebene, (−)-α-muurolene, α-humulene, and a mixture of (−)- and (+)-α-copaene were inhibitory to hydroxylation of (+)-δ-cadinene. In addition, (−)-α-cubebene, (−)-α-muurolene, α-humulene, and, to a smaller extent, (−)-δ-cadinene served as alternative substrates for (+)-δ-cadinene-8-hydroxylase and were converted to mono-hydroxylated products. Of the five olefins tested, α-humulene and α-copaene are found in lysigenous glands of cotton (Elzen et al., 1985), which are also the site of gossypol accumulation ( [Bell et al., 1978] and [Mace et al., 1976]) and the probable site of its biosynthesis.     

Correlated evolution of leaf shape and trichomes in Begonia dregei (Begoniaceae)

Structural features of leaves, including size, shape, and surfaces, vary greatly throughout the plant kingdom. In both functional and phylogenetic analyses of leaves, the various morphological aspects are often considered independently of each other, although it is likely that many combinations of features do not occur at random due to either functional constraint or genetic correlation. The distribution of variation in leaf morphology in the highly variable Begonia dregei species complex was examined in natural populations and in F(2) offspring from a cross between plants from two populations. Leaf shape was quantified using several morphometric measures, and trichomes on leaves were counted and measured. Correlations between leaf shape and the numbers and size of trichomes were examined. There were significant correlations between the shapes of leaves and the presence, number, and size of trichomes among populations and in hybrid plants. Deeply incised leaves had larger numbers of longer trichomes at the sinuses. Higher numbers of trichomes on upper leaf surfaces occurred together with trichomes at the petiole and on the abaxial surface. The potential for independent evolution of leaf shape and trichomes in this group is limited. Hypotheses to explain the correlated development of leaf shape and trichomes are discussed.     

Intra-specific phenotypic variations in Lantana camara leaves affect host selection by the gall maker Aceria lantanae

Leaf galls induced by Aceria lantanae occur in distinct populations of Lantana camara, exclusively on individuals with red flowers. Anatomical, histochemical and phytochemical analyses of leaves were performed to determine characteristics related to host selection among individuals with red (RF), pink (PF) and white flowers (WF). The host plants (RF) presented amphistomatic leaves, higher stomatal index value, and non-glandular trichomes significantly longer and less dense when compared to non host plants. Morphological features of RF indicated a more propitious micro-habitat for A. lantanae, with physical protection against its natural enemies. The monoterpenes limonene and eucalyptol, and the sesquiterpene α-caryophyllene were detected only in non host plants. These differences in volatiles composition might be related to the selection of RF as host plants, and chemical repellence in PF and WF. Our analysis revealed intra-specific morphological and chemical variations possibly related to host selection by A. lantanae.     

Variation in morphology of <Emphasis Type="Italic">Teucrium polium</Emphasis> aggr. populations in Bulgaria

The aim of this study was to investigate natural serpentine and non-serpentine populations of Teucrium polium aggr. and to document the differences in their morphological traits, as well as estimate which characters are most likely contributing to differentiation of the populations. Nine populations distributed both on and off serpentine soils were investigated, and 12 different morphological features of 270 individuals were studied by univariate and multivariate statistical analyses. The results did not demonstrate clear delimitation of the serpentine from the non-serpentine populations. Using these ecotypes as classification factors in the discriminant analysis, it can be stated that stem height, leaf length in the fifth leaf pair, internode length between the fourth and the fifth leaf pairs, and corolla tube length are features with the greatest discriminant ability. The character variation was generally higher for the vegetative features, while reproductive features failed to demonstrate clear differences. The similarity groupings indicated by the cluster analysis were synchronous with groups distinguished by both the discriminant and PCA analyses. Significant differences were observed between sets of vegetative characters in all populations investigated.      

POLLINATION IN FLORAL SCENT MORPHS OF POLEMONIUM VISCOSUM: A MECHANISM FOR DISRUPTIVE SELECTION ON FLOWER SIZE

Plants of Polemonium viscosum have flowers that are either sweet or skunky in scent. The two morphs are preferentially pollinated by insects of strongly contrasting body size: bumblebee queens specialize on sweet flowers, flies on skunky ones. In this study 13 characters were examined in plant specimens from five populations to identify major components of intraspecific variation in flower and inflorescence morphology and test their correlation with floral scent. Factor analysis identified four major axes of morphological variation. The first explained 22% of the variance among specimens and correlated strongly with four flower size characters: sepal length, corolla tube length, corolla lobe width, and corolla lobe length. Floral scent morphs differed significantly in the multivariate representation of flower size defined by these characters. Sweet flowers had wider corolla lobes, longer corolla tubes, and longer sepals than skunky ones. Corolla lobe width accounted for the greatest amount of intermorph divergence. Divergence in flower size between morphs was maintained in mixed populations at four locations in alpine Colorado, with corollas of sweet flowers significantly broader or more flared than those of skunky flowers. Patterns of pollen receipt suggest that this difference is adaptive. In the sweet morph, pollination intensity and purity increased significantly with corolla flare. Conversely, in the skunky morph, corolla flare had little influence on pollination intensity and had a strong negative effect on purity. These findings suggest that selection for effective pollination should favor intraspecific divergence in flower size in Polemonium viscosum.