Preliminary study on the phytochemical evolution of different Lamiaceae species based on iridoids

In this work, an interesting potential application of phytochemistry is reported. In particular, we employed the Sporne diagrams to provide information about the evolution of some different Lamiaceae species in relation to their content in secondary metabolites. In more details, fifteen species belonging to six different genera were studied i.e. Ajuga L., Galeopsis L., Melittis L., Sideritis L., Stachys L. and Teucrium L. The selected secondary metabolites were iridoids because of their wide occurrence and distribution within all the family, thus providing a more accurate and general overview. Nineteen different compounds of this class were identified in the studied species. The aim of this work was to verify if the evolutionary data available in literature for the studied species and based on molecular studies, matched with those obtained from a more phytochemical approach or not. The final objective was to ascertain a correspondence between these two opposite methods. The results of this study clearly showed that, only in some cases, this correspondence was present. This may suggest the possibility to use both methods for drawing phylogenetic trees of plants. Anyway, this study is not general and is quite limited in studied species and in collection areas and does not intend to affirm its absolute accuracy even though it may be a good starting point for future researches in this field.     

Essential oil composition and total metabolite content of a chemotype of Ajuga reptans L. (Lamiaceae) collected in Central Italy

In this work, an interesting application of phytochemistry for ethnopharmacological purposes was reported. In particular, the total phytochemical analysis of a single sample of Ajuga reptans L. (Lamiaceae) collected in Central Italy was carried out in order to provide a rationale for the use of this plant in folk medicine. This analysis was conducted on the volatile oil fraction by gas chromatography–mass spectroscopy (GC-MS) and on the polar one by means of classical column chromatography and nuclear magnetic resonance and MS. Thirty-nine volatile components were identified in the essential oil whereas 32 compounds were identified in the polar fraction. All of these are known to exert several beneficial medicinal activities and no potentially toxic compounds were identified such as the neo-clerodane diterpenoids, commonly evidenced in the entire genus. This study could provide a phytochemical rationale for the possible ethnopharmacological use of this specific species.     

Phytochemical Profiling by UPLC-ESI-QTOF-MS of Kalaharia uncinata (Schinz) Moldenke,Widely Used in Traditional Medicine in DR Congo

Kalaharia uncinata (Schinz) Moldenke, is a tropical erect bushy shrub or subshrub of the Lamiaceae family. It is an endemic plant species of Southern Africa, widely used in the pharmacopoeia against upper respiratory tract infections. A previously conducted ethnobotanical survey revealed that it is believed to contain bioactive substances. However, no relevant phytochemical information was available. This study aimed to perform a phytochemical characterization of K. uncinata and also to discuss the potential bioactivity of the identified phytochemical constituents based on documented data. Ultra-performance liquid chromatography with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS) was used for profiling and identification of the main phytochemical constituents from leaf extracts (MeOH 90 %, DCM, AcOEt, BuOH, hexane and residue) of K.uncinata. Twenty-four constituents, representing mainly flavonoids (14), followed by phenylethanoid glycosides (7), phenolic acids (2), and an iridoid glycoside (1) were tentatively identified. Most of the identified compounds are documented to have antiviral and anti-inflammatory properties, which could possibly be the rationale behind the use of K. uncinata against upper respiratory tract infections.     

Phenotypic and phytochemical diversity among different populations of Stachys lavandulifolia

Stachys lavandulifolia is an herb growing wild in Iran with interesting medicinal properties. In this research, the variability of morphological characters and phytochemical compositions among and within eight populations of this species was studied. Characteristics including inflorescence length, internode length, leaf shape, corolla color, state of calyx trichomes, state of leaf trichomes and corolla length were the most variable morphological characteristics among the examined populations. The phytochemical characterization revealed 45 components in the essential oils. Germacrene D and δ-cadinene were the main component with the highest contents. α-Copaene was positively correlated with δ-cadinene and limonene, while germacrene D was negatively correlated with δ-cadinene. UPGMA dendrogram constructed based on combined data of morphological variables and phytochemical compositions distinguished studied populations with high inter- and intra-population diversity. The high level of phytochemical and morphological variability among and within the studied populations suggests a breeding approach during the domestication, to gain new, promising, and homogenous cultivars, attractive for the industry and agriculture.     

Phylogeny-Aware Chemoinformatic Analysis of Chemical Diversity in Lamiaceae Enables Iridoid Pathway Assembly and Discovery of Aucubin Synthase

Countless reports describe the isolation and structural characterization of natural products, yet this information remains disconnected and underutilized. Using a cheminformatics approach, we leverage the reported observations of iridoid glucosides with the known phylogeny of a large iridoid producing plant family (Lamiaceae) to generate a set of biosynthetic pathways that best explain the extant iridoid chemical diversity. We developed a pathway reconstruction algorithm that connects iridoid reports via reactions and prunes this solution space by considering phylogenetic relationships between genera. We formulate a model that emulates the evolution of iridoid glucosides to create a synthetic data set, used to select the parameters that would best reconstruct the pathways, and apply them to the iridoid data set to generate pathway hypotheses. These computationally generated pathways were then used as the basis by which to select and screen biosynthetic enzyme candidates. Our model was successfully applied to discover a cytochrome P450 enzyme from Callicarpa americana that catalyzes the oxidation of bartsioside to aucubin, predicted by our model despite neither molecule having been observed in the genus. We also demonstrate aucubin synthase activity in orthologues of Vitex agnus-castus, and the outgroup Paulownia tomentosa, further strengthening the hypothesis, enabled by our model, that the reaction was present in the ancestral biosynthetic pathway. This is the first systematic hypothesis on the epi-iridoid glucosides biosynthesis in 25 years and sets the stage for streamlined work on the iridoid pathway. This work highlights how curation and computational analysis of widely available structural data can facilitate hypothesis-based gene discovery.     

Problems in the use of plant biochemistry for establishing the safety of biological control agents for weeds: TheChondrilla andEchium/heliotropium cases

Demonstration of specificity, on the one hand, of the biological control agents for the weedChondrilla juncea L. [Compositae: Cichoriae] as compared to those for the control of the weedsEchium plantagineum L. andHeliotropium europaeum L. (Tubiflorales:Boraginaceae) represents 2 extremes. In the case ofC. juncea the phytochemical data was insufficiently known to be of value and if it had existed, would have been of little value in demonstrating quarantine safety for certain of the agents. However, in the case of the 2Boraginaceae there is strong evidence that studies based on phytochemical data would have confirmed specificity.     

Investigating the medicinal potential,material basis and mechanism of Polygoni Orientalis Fructus based on multi-technology integrated network pharmacology

BackgroundPolygoni Orientalis Fructus (POF) refers to the dried ripe fruit of Polygonum orientale L. which has a long historical application in clinic for treatment of various conditions in China. However, its chemical constituents, pharmacological effects and their coupled correlation have not been intensively investigated.PurposeIn present work, we aimed to elucidate the medicinal material basis, optimum indication and corresponding therapeutic mechanism of POF.MethodsThe main phytochemical ingredients in POF were characterized by liquid chromatography-mass spectrometry (LC-MS) analysis. The optimum medicinal potential and corresponding molecular mechanism of POF were deduced based on integrated statistic pattern recognition and network pharmacology. The deduced pharmacologic efficacy and mechanism of POF were further validated through in vitro study in free-fatty acid (FFA)-induced LO₂ cells.ResultsTotal 30 main phytochemical ingredients were identified in POF in which 18 ingredients were screened to yield 277 potential targets. Based on analyzing the quantitative data matrix of drug-disease targets by statistic pattern recognition, non-alcoholic fatty liver disease (NAFLD) was screened as the optimum indication of POF from 23 candidate diseases. Promising action targets (PPARG, IL6, TNF, IL1B, IKBKB, RELA, etc.) and signaling pathways (AMPK signaling pathway, NF-κB signaling pathway, etc.) were screened and refined to elucidate the therapeutic mechanism of POF against NAFLD based on network pharmacology. In vitro study demonstrated that POF effectively alleviated FFA-induced steatosis, oxidative stress, mitochondrial dysfunction and inflammation, and these beneficial effects were attributed to the activation of AMPK signaling pathway and suppression of NF-κB signaling pathway.ConclusionPOF could be exploited as a promising phytotherapy in the treatment of NAFLD.     

New flavonoids from the underground parts of Eriosema laurentii

Pharmacological, toxicological and phytochemical investigations of aerial parts of Eriosema laurentii De Wild. (Leguminosae), a plant used in traditional African medicine, showed a complex pattern of very rare and several new exceptional phenolic compounds. In continuation of the phytochemical work and based on the activity of an extract from the underground parts, the presented study showed the occurrence of a new pyranoisoflavone, named methyleriosemaone D, and a new highly prenylated (2S,3S)-6,8,3′-triprenyl-3,5,7,2′,4′-pentahydroxyflavanone in these parts of E. laurentii. In addition, the very rare pyranoisoflavone eriosemaone D was proven in this plant for the first time. All structures were elucidated unambiguously by extensive MS- and one and two-dimensional NMR-experiments. Detailed NMR spectra of eriosemaone D were included to rationalize the correction of some of the previously reported carbon signals in the ¹³C NMR spectrum. The estrogenic activity of the extract and isolated compounds was tested in an estrogen receptor α yeast transactivation assay.     

Iridoids from Caryopteris x clandonensis

In continuation of our phytochemical studies on Caryopteris x clandonensis (Lamiaceae), three further iridoids were isolated from the methanolic extract of the stems. Their structures were established by 1D and 2D NMR and MS analysis as a C-6 epimer of 8-O-acetylharpagide (6-epi-8-O-acetylharpagide), a derivative of harpagide which contained the unusual feature of a 3',4' seco-glycopyranosyl moiety (clandonoside II) and a methyl cetal of 8-O-acetylharpagide aglucone hydrate named clandonensine.     

Why Africa matters: evolution of Old World Salvia (Lamiaceae) in Africa

Background and Aims

Salvia is the largest genus in Lamiaceae and it has recently been found to be non-monophyletic. Molecular data on Old World Salvia are largely lacking. In this study, we present data concerning Salvia in Africa. The focus is on the colonization of the continent, character evolution and the switch of pollination systems in the genus.

Methods

Maximum likelihood and Bayesian inference were used for phylogenetic reconstruction. Analyses were based on two nuclear markers [internal transcribed spacer (ITS) and external transcribed spacer (ETS)] and one plastid marker (rpl32-trnL). Sequence data were generated for 41 of the 62 African taxa (66 %). Mesquite was used to reconstruct ancestral character states for distribution, life form, calyx shape, stamen type and pollination syndrome.

Key Results

Salvia in Africa is non-monophyletic. Each of the five major regions in Africa, except Madagascar, was colonized at least twice, and floristic links between North African, south-west Asian and European species are strongly supported. The large radiation in Sub-Saharan Africa (23 species) can be traced back to dispersal from North Africa via East Africa to the Cape Region. Adaptation to bird pollination in southern Africa and Madagascar reflects parallel evolution.

Conclusions

The phenotypic diversity in African Salvia is associated with repeated introductions to the continent. Many important evolutionary processes, such as colonization, adaptation, parallelism and character transformation, are reflected in this comparatively small group. The data presented in this study can help to understand the evolution of Salvia sensu lato and other large genera.     

Effects of Hybridization and Evolutionary Constraints on Secondary Metabolites: The Genetic Architecture of Phenylpropanoids in European Populus Species

The mechanisms responsible for the origin, maintenance and evolution of plant secondary metabolite diversity remain largely unknown. Decades of phenotypic studies suggest hybridization as a key player in generating chemical diversity in plants. Knowledge of the genetic architecture and selective constraints of phytochemical traits is key to understanding the effects of hybridization on plant chemical diversity and ecological interactions. Using the European Populus species P. alba (White poplar) and P. tremula (European aspen) and their hybrids as a model, we examined levels of inter- and intraspecific variation, heritabilities, phenotypic correlations, and the genetic architecture of 38 compounds of the phenylpropanoid pathway measured by liquid chromatography and mass spectrometry (UHPLC-MS). We detected 41 quantitative trait loci (QTL) for chlorogenic acids, salicinoids and flavonoids by genetic mapping in natural hybrid crosses. We show that these three branches of the phenylpropanoid pathway exhibit different geographic patterns of variation, heritabilities, and genetic architectures, and that they are affected differently by hybridization and evolutionary constraints. Flavonoid abundances present high species specificity, clear geographic structure, and strong genetic determination, contrary to salicinoids and chlorogenic acids. Salicinoids, which represent important defence compounds in Salicaceae, exhibited pronounced genetic correlations on the QTL map. Our results suggest that interspecific phytochemical differentiation is concentrated in downstream sections of the phenylpropanoid pathway. In particular, our data point to glycosyltransferase enzymes as likely targets of rapid evolution and interspecific differentiation in the ‘model forest tree’ Populus.     

Does the Arcto-Tertiary Biogeographic Hypothesis Explain the Disjunct Distribution of Northern Hemisphere Herbaceous Plants? The Case of Meehania (Lamiaceae)

Despite considerable progress, many details regarding the evolution of the Arcto-Tertiary flora, including the timing, direction, and relative importance of migration routes in the evolution of woody and herbaceous taxa of the Northern Hemisphere, remain poorly understood. Meehania (Lamiaceae) comprises seven species and five subspecies of annual or perennial herbs, and is one of the few Lamiaceae genera known to have an exclusively disjunct distribution between eastern Asia and eastern North America. We analyzed the phylogeny and biogeographical history of Meehania to explore how the Arcto-Tertiary biogeographic hypothesis and two possible migration routes explain the disjunct distribution of Northern Hemisphere herbaceous plants. Parsimony and Bayesian inference were used for phylogenetic analyses based on five plastid sequences (rbcL, rps16, rpl32-trnH, psbA-trnH, and trnL-F) and two nuclear (ITS and ETS) gene regions. Divergence times and biogeographic inferences were performed using Bayesian methods as implemented in BEAST and S-DIVA, respectively. Analyses including 11 of the 12 known Meehania taxa revealed incongruence between the chloroplast and nuclear trees, particularly in the positions of Glechoma and Meehania cordata, possibly indicating allopolyploidy with chloroplast capture in the late Miocene. Based on nrDNA, Meehania is monophyletic, and the North American species M. cordata is sister to a clade containing the eastern Asian species. The divergence time between the North American M. cordata and the eastern Asian species occurred about 9.81 Mya according to the Bayesian relaxed clock methods applied to the combined nuclear data. Biogeographic analyses suggest a primary role of the Arcto-Tertiary flora in the study taxa distribution, with a northeast Asian origin of Meehania. Our results suggest an Arcto-Tertiary origin of Meehania, with its present distribution most probably being a result of vicariance and southward migrations of populations during climatic oscillations in the middle Miocene with subsequent migration into eastern North America via the Bering land bridge in the late Miocene.     

Q-TOF LC-MS compounds evaluation of propolis extract derived from Malaysian stingless bees,Tetrigona apicalis,and their bioactivities in breast cancer cell,MCF7

Propolis is known to exhibit various phytochemical compounds that aid in several biological activities. The current study investigates the phytochemical compounds of ethanolic extract of propolis of Tetrigona apicalis (EEP) using Q-TOF LC-MS, its antioxidant properties using DPPH and ABTS⁺ radical scavenging assays, total phenolic (TPC) and flavonoid content (TFC), using Folin-Ciocalteu and Aluminium Chloride method, respectively, as well as proapoptotic effects, based on the selected IC₅₀ of the cytotoxic study conducted for EEP using annexin V-FITC assay. Terpene and polyphenol were among of 17 identified compounds. The EC₅₀ of EEP for DPPH and ABTS⁺ was 1.78 mg/mL and 1.68 mg/mL, while the EEP exhibited TPC and TFC values of 31.99 mgGAE/g and 66.4 mgQCE/g, respectively in which the parameters were strongly correlated. The IC₅₀ of EEP effectively induces apoptosis in MCF7 cells. In conclusion, EEP possessed important phytochemical compounds that work excellently as antioxidants and anticancer agents.     

Lapachol and its congeners as anticancer agents: a review

Lapachol is a naturally occurring 1,4-naphthoquinone originally isolated by the Italian phytochemist E. Paterno from Tabebuia avellanedae (Bignoniaceae) in 1882 and subsequently found in several other genera belonging to the families of Leguminosae, Malvaceae, Plumbaginaceae, Lamiaceae, Arecaceae, Scrophulariaceae, Verbenaceae, Celastraceae, Avicenniaceae, Caesalpiniaceae, Rubiaceae, and Proteaceae. A wide range of pharmacological activities have been observed for lapachol and its semi-synthetic derivatives in the literature, such as antileishmanial, anticarcinomic, anti-inflammatory, antimalarial, antiseptic, antitumor, antiviral, bactericidal, fungicidal, insectifugal, pesticidal, schistosomicidal, termiticidal, and viricidal effects. The aim of this review is to discuss in detail the phytochemical properties and pharmacological effects of the title compound that have been reported thus far, highlighting its potential therapeutic benefits for the future.     

Isolation and structural elucidation of two new labdane diterpenoids from the aerial part of Roylea cinerea

Two new labdane diterpenoids, cinereanoid A (1) and cinereanoid B (2), along with five known compounds, calyone (3), pilloin (4), 1-methylindole-3-carboxaldehyde (5), β-sitosterol (6) and stigmasterol (7) were isolated from the aerial parts of Roylea cinerea (Lamiaceae). The new structures were determined by using IR, MS, 1D, 2D NMR spectroscopy. The structure of both new compounds was further confirmed by single crystal X-ray crystallographic analysis. In this study we have also reported single crystal X-ray structure of compound 3 which unambiguously confirmed the relative stereochemistry of tertiary hydroxyl and methyl groups, as it was not established by earlier report. Compounds 4 and 5 were isolated for the first time from this plant. In view of very few reports about this species, this report has increased the phytochemical knowledge about R. cinerea.     

Molecular phylogeny of Phalaenopsis Blume (Orchidaceae) on the basis of plastid and nuclear DNA

Representatives of the genus Ancistrocladus, woody lianas of tropical Africa and Asia, contain pharmaceutically interesting alkaloids and have been the subjects of intensive phytochemical investigations. In Southeast Asia, Ancistrocladus tectorius, previously regarded as the only species of the genus from this region, is extremely polymorphic with respect to naphthylisoquinoline alkaloids, indicating that this taxon might be comprised of several morphologically similar species. We performed a comparative study of the ITS region of nuclear rDNA and of the trnK intron of cpDNA as well as an ISSR fingerprint analysis. Using 75 samples of A. tectorius from 21 locations in comparison to samples of other Ancistrocladus species from Asia and from West and Central Africa, we investigated patterns of species differentiation within this taxon. We found the high variability of chemical compounds described for A. tectorius to be paralleled by a high genetic variability of the units that have been assigned to this taxon. Samples assigned to A. tectorius were paraphyletic with respect to species from the Indian subcontinent, and intraspecific variability was comparable to interspecific variability among the African taxa. In addition, groups of individuals occurring in sympatry were found to be more similar to those from other locations, suggesting low levels of gene flow between those sympatric groups. This indicates either a considerable number of hybridization events during the evolution of A. tectorius or the existence of several distinguishable species not yet recognized. Our results are a first step in the development of species or population-specific markers for the prediction of the alkaloid spectrum of samples. This will help to improve the reproducibility of phytochemical research on Ancistrocladus.     

Terpenoids and a coumarin from aerial parts of Psychotria vellosiana Benth. (Rubiaceae)

The phytochemical investigation of Psychotria vellosiana aerial parts yielded five compounds, including two triterpenes (squalene and lupeol), two steroids (stigmasterol and sitosterol), and one coumarin (scopoletin). The structures of the isolates were established based on analysis of their spectroscopic data and comparison with literature. All metabolites isolated were previously detected in Psychotria, excepting squalene that is being described for the first time from this genus. This study adds a new species to a number of reports showing the absence of alkaloid in Psychotria.     

Variability of qualitative and quantitative secondary metabolites traits among wild genetic resources of Lavandula stoechas L.

Lavandula stoechas L. (Lamiaceae) is an attractive shrub native to the Mediterranean regions used for ornamental, melliferous, aromatic and medicinal purposes. Furthermore, this species presents an increasing interest in cosmetics, perfumery and pharmaceutical industries. The variability of qualitative and quantitative metabolic traits among nine wild germplasms representing the distribution area of this species in Tunisia was undertaken. A total of 45 essential oil components were identified in the aerial parts of the studied germplasms. The main essential oil components were camphor (15.32–50.63%), fenchone (6.57–34.70%), 1,8-cineole (0.05–13.45%) and γ-gurjunene (1.10–12.15%). In addition to the well known chemotypes camphor/fenchone and camphor/1.8-cineole, a new chemotype camphor/γ-gurjunene was detected in Tunisian L. stoechas L. Six phenolic acids (quinic acid, gallic acid, p-coumaric acid, 4,5-di-O-caffeoyquinic acid, salviolinic acid and trans cinnamic acid) and five flavonoids (luteolin-7-o-glucoside, naringin, apegenin-7-o-glucoside, quercetin and kampherol) were identified in the ethanolic extracts. Salviolinic acid (46.30–615.18 μg/g) and luteolin-7-o-glucoside (5.98–38.54 μg/g) were the most abundant phenolic compounds. A high significant phytochemical variability (p ˂ 0.01) in the accumulation of volatile and phenolic secondary metabolites among the studied germplasms was recorded. The conducted multivariate (PCA) and clustering (HCA) analyses revealed different classification pattern for essential oil and phenolic compounds. The detected phytochemical polymorphism among the investigated lavender ecotypes didn't show accordance with bioclimatic and geographical areas which suggests genetic background as the main explaining factor. The detected secondary metabolites polymorphism valorises Tunisian L. stoechas L. genetic resources as valuable plant material in further breeding programs. Moreover, an urgent in situ and ex situ conservation measures are required for these wild germplasms threatened by human over-harvesting practices and the occurring dramatic changes in climatic conditions.     

Himalsamines A and B,two new amino sugar isomers from the leaves of Daphniphyllum himalaense subsp. macropodum

A phytochemical investigation performed on the leaves of Daphniphyllum himalaense has led to the characterization of two new amino sugars (1 and 2) and six known daphniphyllum alkaloids. The structures of new compounds were determined based on the interpretation of spectroscopic data. Some of the alkaloids were found to possess moderate to weak activity against the hepatitis C virus infection.     

Phytochemical study on the leaves of Wollemia nobilis

In this work, a phytochemical study performed on the leaves of the rare species, Wollemia nobilis W.G. Jones, K.D. Hill & J.M. Allen, is reported. By means of classical column chromatography and NMR Spectroscopy and Mass Spectrometry, nine compounds were evidenced. These were: pheophorbide a (1), isocupressic acid (2), acetyl-isocupressic acid (3), sandaracopimaric acid (4), agathic acid (5), 7–4′-4‴-tri-O-methyl-agathisflavone (6), 7–4′-7″-4‴-tetra-O-methyl-agathisflavone (7), caffeic acid (8) and shikimic acid (9). Compared to our previous phytochemical analysis on the male cones, some further compounds were identified i.e. compounds 1, 5, 6, 7 and 8. This confirmed the previous chemotaxonomic considerations of the species but also added new ones which were discussed within the text. In addition, a possible different accumulation of secondary metabolites in the tissues and organs of this plant was even noticed.