Analysis of polyphenols,phytosterols, and bitter principles in <Emphasis Type="Italic">Teucrium</Emphasis> L. species

Total polyphenols, tannins, β-sitosterol, and bitterness values were determined in native and cultivated populations of Teucrium L. species from Croatia (T. arduini L., T. botrys L., T. chamaedrys L., T. flavum L., T. montanum L., T. polium L., and T. scordium L. subsp. scordioides Schreb.). Teucrium species have long been present in folk medicine for diverse medicinal uses, but little is known about polyphenols, phytosterols, and bitter principles. Spectrophotometrically determined contents of total polyphenols (TP) and tannins (T) significantly varied among investigated Teucrium species and were somewhat higher in native populations. The highest TP and T contents were measured in native specimens of T. montanum (TP: 13.68%; T: 3.48%). Scanning densitometry was used for measurements of β-sitosterol levels in plant samples ranged from 0.056% (T. montanum) to 0.129% (T. botrys), and it was not significantly different between native and cultivated specimens of the same plant species. Bitterness values were similar for native and cultivated samples of the same plant species; the highest was measured for T. montanum (15659). The present study suggests that Teucrium species growing in Croatia have potential for cultivation and might be a valuable source of natural bioactive compounds.     

Morphology and distribution of trichomes in Italian species ofTeucrium sect.Chamaedrys (Labiatae) — a taxonomical evaluation

Micromorphology and distribution of trichomes (both secretory and not) in different parts of plants of the five Italian species belonging toTeucrium sect.Chamaedrys have been studied. These characters neatly evidence the differences between the taxa considered; of particular interest are the observations concerning the dense coverage of peltate hairs on the abaxial face of the leaves ofT. flavum subsp.glaucum and the characters contributing to a better identification ofT. marum andT. subspinosum. The importance of the calyx indumentum as distinctive taxonomical character of the genusTeucrium is confirmed, stressing at the same time the necessity of completing the observations at micromorphological level.     

Anatomical and nutlet differentiation between <Emphasis Type="Italic">Teucrium montanum</Emphasis> and <Emphasis Type="Italic">T. polium</Emphasis> from Turkey

Teucrium montanum L. and T. polium L. are the two closest Teucrium L. species from sect. Polium (Mill.) Schreb in Turkey. In addition, they are sympatric for some part of their range in Turkey. In this study, comparative anatomical and micromorphological studies of the two species are carried out. They have been investigated by their leaf and stem anatomical features, as well as nutlet micromorphological characteristics. The results of anatomical studies show that the anatomical characters of both taxa are observed to be similar to the general features of the family Lamiaceae anatomy, except for lacking rich collenchyma at the corners. Both taxa are similar in stem anatomy and their leaves exhibit xeromorphy. However, trichome morphology on the stems and the leaves appear to have a taxonomic value in segregation of the two taxa. Light and scanning electron microscope studies on the nutlets also show that nutlet shapes, measures and surface micromorphologies are different in the two species. Whereas nutlet surfaces are bireticulate in both species, the nutlets are larger and primary sculpturing is more distinct in T. polium than in T. montanum. Moreover, the nutlets are oblong to oblong-ovoid and larger in T. polium, but ovoid and smaller in T. montanum.     

Antimicrobial activity,total phenolic content and flavonoid concentrations of <Emphasis Type="Italic">Teucrium</Emphasis> species

In vitro antimicrobial activity of 21 crude extracts obtained from seven taxa of the genus Teucrium (T. chamaedrys, T. montanum, T. arduini, T. polium, T. scordium subsp. scordium, T. scordium subsp. scordioides and T. botrys) was tested against bacterial and fungal species. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined using a microdilution analysis method. Total phenolic content and flavonoid concentrations were measured spectrophotometrically. Total phenols were determined using Folin-Ciocalteu reagent and their amounts ranged from 28.49 up to 159.84 mg CA/g of extract (chlorogenic acid equivalent). The amounts of flavonoids ranged from 38.17 up to 190.45 mg RU/g of extract (rutin equivalent).The plant extracts showed greater potential of antibacterial than antifungal activity. A relationship was found between total phenolics and biological activity. The highest level of total phenols was measured in the methanol extracts, which demonstrated higher antimicrobial activity than acetone and ethyl acetate extracts. Staphylococcus aureus ATCC 25923 appeared to be the most sensitive organism. Our results indicate that Teucrium spp extracts are rich sources of phenolic compounds and are promising candidates for further development as natural antimicrobial agents.     

Clerodane diterpenoids from Teucrium and Ajuga plants

A new clerodane diterpenoid, chamaepitin has been isolated from Ajuga chamaepitys. The previously known clerodanes, 19-acetylgnaphalin, auropolin, teucrin A and teuflin, have been isolated from Teucrium species (T. polium subsp. belion, T. polium subsp. capitatum and T. asiaticum) collected in Majorca. X-ray diffraction analysis on crystals of auropolin has allowed the assignment of configuration at C-20 as S for this compound.     

Biological activities of Teucrium flavum L., Teucrium fruticans L., and Teucrium siculum rafin crude extracts

The genus Teucrium (Lamiaceae) includes 300 species widespread all around the world, which are perennial herbs or shrubs commonly, named germanders. In Italy, Teucrium flavum L., Teucrium fruticans L., and Teucrium siculum Rafin are mostly present in Liguria, Sicily, and Sardegna. Teucrium species are characterized by mono and sesquiterpene hydrocarbons, flavonoids, fatty acid esters, and essential oils. Many species of this genus show antioxidant, antimicrobial, and antifungal activities, rendering them useful as natural preservative ingredients. In view of the interesting biological properties reported for Teucrium spp., in this study we determined the total phenol and flavonoid content of inflorescence extracts of T. flavum L., T. fruticans L., and T. siculum Rafin. In addition, we investigated the in vitro antioxidant and antibacterial activities of inflorescence extracts against pathogenic bacteria. Obtained results demonstrated that extracts had in vitro antioxidant activity and showed antimicrobial ability against Gram-positive and Gram-negative strains albeit with different effectiveness probably due to the different qualitative/quantitative composition of the extract also suggesting that these extracts might be useful in preventing several diseases in which oxidative stress may represent an important pathogenic mechanism.     

A systematic revision of the genus Teucrium (Lamiaceae) in Egypt

A critical taxonomic revision of the genus Teucrium (Lamiaceae) in Egypt is presented. It reveals the presence of five species, including two subspecies and four varieties. Two new nomenclatural combinations are proposed: Teucrium jordanicum and T. jordanicum var. sinaicum. The latter in addition to T. polium subsp. capitatum are added for the first time to the flora of the country. A key, annotated enumeration of species and notes on the distribution of the species are also provided.     

Establishing the genetic relationships between the wild and cultivated olives using a nuclear intron from nitrate reductase (nia-i3)

In Oleaceae the most outstanding biological issue is to clarify the taxonomic relationships of cultivated and wild olives. To establish the genetic relationships between the wild (Olea europaea subsp. europaea var. sylvestris (Mill.) Lehr.), the cultivated olive (Olea europaea subsp. europaea var. europaea), and other taxa of the genus Olea (Olea europaea subsp. cuspidata (Wall. ex G. Don) Cif., Olea europaea subsp. cerasiformis G. Kunkel & Sunding, Olea paniculata R. Br.) and other Oleaceae (represented by Ligustrum vulgaris) we carried out the amplification by polymerase chain reaction (PCR) and the sequencing of the third nuclear intron of the nitrate reductase gene (nia-i3). Sequence analyses showed the presence of two different functional variants of the intron (nia1 and nia2) in the Oleaceae, in addition to a shorter non-functional one. Notably, while the shortest and the nia1 variants were present in all the taxa analysed, the nia2 variant was present only in the wild and the cultivated olive. These data confirm the close phylogenetic relationship between wild and cultivated olives and suggest that this gene could be duplicated in these two taxa after its divergence from the remaining Oleaceae. The presence of a target for AflII enzyme in nia2 and its absence in nia1 variant enables easy distinction by PCR-RFLP between, on the one hand, wild and cultivated olive, and on the other the remaining subspecies of the Olea europaea L. complex (O. e. subsp. cuspidata and O. e. subsp. cerasiformis) as well as other Oleaceae (O. paniculata, L. vulgaris L.). Additionally, nia1 sequences provide useful information about phylogeny of the wild and cultivated olives inside the genus Olea.     

Biodiversity characteristics of Teucrium polium species in Saudi Arabia

Teucrium (Lamiaceae) is a large and polymorphic genus distributed mainly in Europe, North Africa and in the temperate parts of Asia. In this study, the anatomical features of the leaf and stem of Teucrium polium are investigated. Teucrium has 19 taxa in Iran, which mainly grow in the Irano-Turanian region between 700 and 2000 m above sea level. T. polium belonging to sect. Polium, is a perennial herb growing on Lorestan province. The leaves clearly exhibit xeromorphy due to features such as thick cuticle layer, thick outer epidermal cell wall, high density of trichomes and thick palisade layer of the mesophyll. Anatomical studies on T. polium revealed that the stem shares the general characteristics of the Labiatae family. The aim of our approach was to study the morphological and taxonomical parameters for T. polium in Saudi Arabia. The results of this study showed that there was no influence of environment on the structure of stomata and trichomes as studying species with T.polium. In conclusion our study shows we have studied the geographical distribution of the species in Saudi Arabia and in the world.     

Comparative study of antioxidant capacity in organs of different Allium species

Cultivated and wild Allium species were investigated in order to compare the antioxidant capacity of their leaves and bulbs. Leaf and bulb anti-oxidative enzymes and scavenger activities, along with quantities of non-enzymic antioxidants, malonyl-dialdehyde and OH radicals were determined. Results obtained suggest that leaves possess higher anti-oxidant and scavenging activities than bulbs in the majority of cultivated and wild Allium species examined. Cultivation of some wild species such as A. flavum L., A. schoenoprasum L. and A. ursinum L. could be conducted in the future in order to produce Allium species with high antioxidant capacity in leaves and bulbs.     

<Emphasis Type="Italic">In vitro</Emphasis> propagation and secondary metabolites production in wild germander (<Emphasis Type="Italic">Teucrium polium</Emphasis> L.)

A protocol for in vitro propagation of the wild germander (Teucrium polium L.) was developed. In vitro plants were developed from ex vitro axillary buds. Then, shoot tips were excised and established on Murashige and Skoog medium. Proliferation of shoots was tested with different levels of 6-furfurylaminopurin, 6-benzyladenine, or thiadiazuron. The highest proliferation of T. polium was obtained when 6-benzyladenine and 6-furfurylaminopurin were used at 2.0 and 1.6 mg l⁻¹, respectively. Thiadiazuron gave the lowest response for shoot proliferation. Rooting was experimented at different levels of Indol-3-butric acid, Indol-3-acetic acid, or 1-naphthaleneacetic acid. 1-Naphthaleneacetic was the only growth regulator which promoted root induction. Rooted plants were acclimatized successfully with 75% survival and grown in the greenhouse. In vitro- and in vivo-grown plants were analyzed for essential oil production. In vitro-grown T. polium on MS medium supplemented with 6-benzyladenine and 1-naphthaleneacetic gave higher oil yield than that grown on hormone-free Murashige and Skoog medium. In vivo (wild)-grown T. polium produced different oil yield when collected in different months (April and October). β-caryophyllene, used as a marker compound in the essential oil, was identified and quantified by gas chromatography (GC) analysis. Gas chromatography/mass (GC-MS) spectrometry analysis was also used to identify other components of in vitro cultures and to compare with in vivo-grown plants.     

Micromorphological differences between some European and American <Emphasis Type="Italic">Fraxinus</Emphasis> (<Emphasis Type="Italic">Oleaceae</Emphasis>) species

Micromorphological differences in leaves and pollen between two American (Fraxinus americana L., F. pennsylvanica Marshall) and two European (F. angustifolia Vahl, F. excelsior L.) ash species were studied using scanning electron microscope. The types, dimensions and distribution of characteristic trichomes were established and measured. Capitate hairs on the leaves had the same shape in all researched ash species. Acicular hairs were regularly present in two American ash species, but very rarely in the glabrous phase of F. angustifolia and F. excelsior. Only F. americana had coronulate abaxial surface of leaves. Pollen of F. angustifolia and F. excelsior had 3 (tricolpate) apertures, and F. americana and F. pennsylvanica 4 (stephanocolpate) apertures. Based on the appearance of the reticulum it’s possible to clearly distinguish all four species. F. angustifolia and F. pennsylvanica had muri with transversal ridges and seldom granules. Muri of F. excelsior and F. americana had slightly visible transversal ridges, and because of that noticeable granules.     

Morphometric variation inLomelosia crenata (Dipsacaceae)

Morphometric characters on 118 herbarium specimens ofLomelosia crenata subsp.crenata (regarded as including subsp.pseudisetensis) were statistically evaluated to detect discontinuities in the variation pattern. Three morphotypes were detected: specimens with short stems and long calyx bristles, specimens with short stems and short bristles, and specimens with long stems and short bristles. The first morphotypic group is separated from the others, which are partly overlapping. These groups broadly correspond toL. crenata subsp.crenata, toScabiosa crenata var.glabriuscula, and toL. crenata subsp.pseudisetensis, respectively.     

Selected chromosome counts of the Czechoslovak flora III

Chromosome numbers compared with as yet published data are given for the following 12 Phanerogams (both native species and aliens) from Czechoslovakia:Ambrosia trifida L.,Cardamine chelidonia L.,Dephne cneorum L.,Epipactis albensis Nováková etRydlo,Linum flavum L.subsp flavum, Lunaria rediviva L.,Nepeta grandiflora M.BIEB.,Reseda luteola L.,Thlaspi montanum L.,Tithymalus salicifolius (Host)Klotzsch etGarcke,Tithymalus virgultosus (Klokov) Holub andVerbascum speciosum Schrad. subsp.speciosum. The chromosome number 2n=40 is presented for the first time in autogamousEpipactis albensis Nováková etRydlo. New chromosome numbers were found inCardamine chelidonia L. (2n=32) and inTithymalus salicifolius (Host) Klotzch etGarcke (2n=40). Known but less frequent cytotypes are reported inLinum flavum L. subsp.flavum (2n=28) and inVerbascum speciosum Schrad. subsp.speciosum (2n=30).     

Population structure and genetic diversity distribution in wild and cultivated populations of the traditional Chinese medicinal plant Magnolia officinalis subsp. biloba (Magnoliaceae)

Magnolia officinalis subsp. biloba, a traditional Chinese medicinal plant, experienced severe declines in the number of populations and the number of individuals in the late 20th century due to the widespread harvest of the subspecies. A large-scale cultivation program was initiated and cultivated populations rapidly recovered the loss in individual plant numbers, but wild populations remained small as a consequence of cutting. In this study, the levels of genetic variation and genetic structure of seven wild populations and five domestic populations of M. officinalis subsp. biloba were estimated employing an AFLP methodology. The plant exhibited a relatively high level of intra-population genetic diversity (h = 0.208 and H _j = 0.268). The cultivated populations maintained approximately 95% of the variation exhibited in wild populations, indicating a slight genetic bottleneck in the cultivated populations. The analysis of genetic differentiation revealed that most of the AFLP diversity resided within populations both for the wild group (78.22%) and the cultivated group (85.92%). Genetic differentiation among populations in the wild group was significant (F _ST = 0.1092, P < 0.005), suggesting wild population level genetic structure. Principal coordinates analysis (PCO) did not discern among wild and cultivated populations, indicating that alleles from the wild population were maintained in the cultivated gene pool. Results from the present study provide important baseline data for effectively conserving the genetic resources of this medicinal subspecies.     

Trichome morphology provides phylogenetically informative characters for Tremandra, Platytheca and Tetratheca (former Tremandraceae)

Summary Trichomes of Tremandra R.Br. ex DC., Platytheca Steetz and Tetratheca Sm. (Elaeocarpaceae, former Tremandraceae), together with two outgroup species of Elaeocarpus L., are illustrated using scanning electron microscopy, and their distribution on various plant organs is documented. Various trichomes types were identified that relate taxa: simple hairs, stellate hairs, short glandular trichomes, long glandular trichomes, and three forms of tubercules. Both outgroup and ingroup taxa have simple hairs. Stellate hairs are confirmed as unique to Tremandra. Prominent and sculptured multicelled tubercules, some bearing a stout hair, are characteristic of Platytheca. Smaller multicelled tubercules occur in both Platytheca and Tetratheca, except for the Western Australian taxon Te. filiformis Benth. (possibly plesiomorphic). Unicellular tubercules (papilla) characterise two species of Tetratheca. Short glandular trichomes, usually found on the ovary, also occur in both of these genera but not in all species (possibly secondary losses), while long glandular trichomes, usually on stems and leaves, occur only in some groups of Tetratheca. Within Tetratheca, Western Australian taxa that have five-merous flowers fall into three ‘groups’: seven species (together with one from South Australia) that have short glandular trichomes but no long glandular trichomes; six species that have long glandular trichomes but no short glandular trichomes; and four species that have both trichome types. All other species of Tetratheca have four-merous flowers and form two ‘groups’: 12 eastern species (including one from South Australia) that have both short glandular trichomes and long glandular trichomes; 4 western species and six eastern species that lack short glandular trichomes. On the basis of these characters, a phylogenetic hypothesis for the three genera is presented.     

Micromorphological studies on Verbascum (Scrophulariaceae) in Iran with emphasis on seed surface, capsule ornamentation and trichomes

Seed micromorphology in 22 species, capsule surface in 28 species and capsule hairs in 20 species of genus Verbascum distributed in Iran were studied using scanning electron microscopy. The seed surface is ridged or alveolate among the studied species. Depth and density of the alveoli or ridges, size of polygonal cells along with arrangement of vesicles on the walls separating the polygonal cells on the seed surface seem to be species-specific in Verbascum. The capsule surface is striate, distinctly or indistinctly, or even smooth in Verbascum. The capsules are rarely glabrous (in V. orientale and V. intricatum), densely covered by branched hairs, or loosely by glandular hairs, depending on species. From the results of this study it is obvious that no correlation exists between the studied species concerning the micromorphological aspects studied here.     

Role of mycorrhizal infection in the growth and reproduction of wild vs. cultivated plants

Summary We tested the hypothesis that mycorrhizal infection benefits wild plants to a lesser extent than cultivated plants. This hypothesis stems from two observations: (1) mycorrhizal infection improves plant growth primarily by increasing nutrient uptake, and (2) wild plants often possess special adaptations to soil infertility which are less pronounced in modern cultivated plants. In the first experiment, wild (Avena fatua L.) and cultivated (A. sativa L.) oats were grown hydroponically at four different phosphorus levels. Wild oat was less responsive (in shoot dry weight) to increasing phosphorus availability than cultivated oat. In addition, the root: shoot ratio was much more plastic in wild oat (varying from 0.90 in the low phosphorus solution to 0.25 in the high phosphorus solution) than in cultivated oat (varying from 0.44 to 0.17). In the second experiment, mycorrhizal and non-mycorrhizal wild and cultivated oats were grown in a phosphorus-deficient soil. Mycorrhizal infection generally improved the vegetative growth of both wild and cultivated oats. However, infection significantly increased plant lifespan, number of panicles per plant, shoot phosphorus concentration, shoot phosphorus content, duration of flowering, and the mean weight of individual seeds in cultivated oat, while it had a significantly reduced effect, no effect, or a negative effect on these characters for wild oat. Poor positive responsiveness of wild oat in these characters was thus associated with what might be considered to be inherent adaptations to nutrient deficiency: high root: shoot ratio and inherently low growth rate. Infection also increased seed phosphorus content and reproductive allocation.     

Ligularia zhengyiana sp. nov. (Asteraceae) from the Hubei Province,China

The new species Ligularia zhengyiana from the Hubei Province, China, is described and illustrated. It is characterized by dense blackish purple pilose hairs on the stems and by coarsely triangular‐dentate leaves. Ligularia zhengyiana is similar to L. fischeri (Ledeb.) Turcz., but L. zhengyiana has dense long blackish purple hairs on stems, while L. fischeri is glabrous or has short yellowish‐brown hairs on stems. In addition, the pappus of L. zhengyiana is white, while that of L. fischeri is yellowish‐brown.