Metabolomic profiling and biological properties of six Limonium species: novel perspectives for nutraceutical purposes.

The genus Limonium includes important halophyte plants containing a variety of bioactive compounds of therapeutic interest. In the present work, the untargeted phytochemical profiles of both aerial part and root extracts from six Limonium species namely, L. bellidifolium, L. globuliferum, L. gmelinii, L. lilacinum, L. sinuatum and L. iconicum from Turkey were determined. Furthermore, several biological activities (in vitro antioxidant and enzyme inhibitory effects) were investigated. Overall, significant amounts of total phenolics (43.64-238.18 mg g-1) and flavonoids (1.61-129.69 mg g-1) were recorded. Particularly, the root extracts of L. gmelinii, L. iconicum and L. globuliferum showed the highest total phenolic content (204.13-238.18 mg g-1), whilst the highest total flavonoid content was recorded in the root extracts of L. gmelinii (129.69 mg g-1). Overall, the tested extracts demonstrated potent radical scavenging activities in both DPPH (2,2- diphenyl-1-picrylhydrazyl) and ABTS (3-ethylbenzothiazoline-6-sulphonic acid) (90.10-507.94 mg g-1 and 163.39-1175.34 mg g-1, respectively). However, the highest scavenging potential (p < 0.05) was displayed by the root extracts of L. iconicum. Conversely, the metal chelating ability assay revealed that L. lilacinum root extract showed the highest activity (21.03 mg g-1). Interestingly, all the extracts were found to be active inhibitors of cholinesterases (AChE (acetylcholinesterase): 4.20-5.11 mg GALAE (galantamine equivalent) per g; BChE (butyrylcholinesterase): 3.89-10.75 mg GALAE per g), amylase (0.52-1.09 mmol ACAE (acarbose equivalent) per g) and tyrosinase (119.41-155.67 mg KAE (kojic acid equivalent) per g), unlike for glucosidase (2.31-2.41 mmol ACAE per g). Taken together, these findings demonstrated a diverse chemical profiles and biological of the extracts, to be potentially considered as phytotherapeutic or functional ingredients due to their antioxidant properties and inhibition of key enzymes involved in several diseases.

In silico pharmacodynamics, toxicity profile and biological activities of the Saharan medicinal plant Limoniastrum feei

ABSTRACT In-silico study was performed to find the pharmacodynamics, toxicity profiles and biological activities of three phytochemicals isolated from Limoniastrum feei (Plumbagenaceae). Online pharmacokinetic tools were used to estimate the potential of Quercetin, kaempferol-3-O-ß-D-glucopyranoside (astragalin) and quercitin-7-O-ß-D-glucopyranoside as specific drugs. Then the prediction of potential targets of these compounds were investigated using PharmMapper. Auto-Dock 4.0 software was used to investigate the different interactions of these compounds with the targets predicted earlier. The permeability of quercetin was found within the range stated by Lipinski ׳s rule of five. Hematopoietic prostaglandin (PG) D synthase (HPGDS), farnesyl diphosphate synthetase (FPPS) and the deoxycytidine kinase (DCK) were potential targets for quercetin, astragalin and quercetin 7, respectively. Quercetin showed antiallergic and anti-inflammatory activity, while astragalin and quercetin 7 were predicted to have anticancer activities. The activity of Astragalin appeared to be mediated by FPPS inhibition. The inhibition of DCK was predicted as the anticancer mechanisms of quercetin 7. The compounds showed interesting interactions and satisfactory binding energies when docked into their targets. These compounds are proposed to have activities against a variety of human aliments such as allergy, tumors, muscular dystrophy, and diabetic cataracts.

Taxonomic complexity in the halophyte Limonium vulgare and related taxa (Plumbaginaceae): insights from analysis of morphological, reproductive and karyological data.

BACKGROUND AND AIMS: Limonium is a well-known example of a group of plants that is taxonomically complex due to certain biological characteristics that hamper species' delineation. The closely related polyploid species Limonium vulgare Mill., L. humile Mill. and L. narbonense Mill. are defined species and can be used for studying patterns of morphological and reproductive variation. The first two taxa are usually found in Atlantic Europe and the third in the Mediterranean region, but a number of intermediate morphological forms may be present alongside typical examples of these species. This study attempts to elucidate morphological, floral and karyological diversity representative of these taxa in the Iberian Peninsula. METHODS: The extent of morphological differentiation was tested through comparison of 197 specimens from both Portugal and Spain using 17 descriptive morphological characters and 19 diagnostic morphometric characters. Analyses of floral morphisms (heterostyly and pollen-stigma dimorphism) and karyological determinations were also conducted. KEY RESULTS AND CONCLUSIONS: Discriminant analysis using morphometric variables reliably assigned individuals in natural populations to their respective groups. In addition, the results provide the first direct evidence that L. narbonense and a new species, LIMONIUM MARITIMUM: Caperta, Cortinhas, Paes, Guara, Espírito-Santo and Erben, SP NOV: , related to L. vulgare are present on Portuguese coasts. Most of these species are found together in mixed populations, especially L. vulgare and L. narbonense. It is hypothesized that taxonomic biodiversity found in sites where distinct species co-occur facilitates the evolutionary processes of hybridization, introgression and apomixis. This study therefore contributes to the elucidation of the taxonomic diversity in L. vulgare-related species and may also help in implementing future conservation programmes to maintain the evolutionary processes generating biodiversity.

Metal phytoremediation by the halophyte Limoniastrum monopetalum (L.) Boiss: two contrasting ecotypes.

The phytoremediation potential of the halophyte Limoniastrum monopetalum for the removal of Cd and Pb from polluted sites is assessed in this work. Two pot experiments were conducted; the first with wild L. monopetalum grown on soil polluted with Cd and Pb irrigated at different salinities, and the second with commonly cultivated ornamental L. monopetalum grown on soil polluted with Cd irrigated also at different salinities. The data revealed that wild L. monopetalum is a Cd and Pb tolerant plant able to accumulate at least 100 ppm of cadmium in its shoots without showing any significant decrease in terms of biomass production, chlorophyll content or water content suggesting that it could be an accumulator of Cd. Pb above-ground accumulation was kept at low levels with the majority of Pb localized in the roots. On the other hand, contrasting results were obtained for ornamental L. monopetalum which although it was found to be also Cd tolerant, Cd accumulation in its tissues was kept at significantly lower levels especially compared to that of the wild ecotype. In addition for ornamental L. monopetalum salinity did not have a positive effect on Cd accumulation and translocation as observed in the wild type and in other halophytes. Analysis of the salt excretion crystals on the leaf surface confirmed that wild and cultivated ornamental L. monopetalum excrete cadmium and lead through their salt glands as a possible metal detoxification mechanism, although the amount excreted by the ornamental L. monopetalum is significantly less.

Comparative study of three Plumbago L. species (Plumbaginaceae) by microscopy, UPLC-UV and HPTLC.

This paper presents a comparative study of anatomy of leaves, stems and roots of three species of Plumbago, namely P. auriculata Lam., P. indica L. and P. zeylanica L. by light microscopy. The paper also provides qualitative and quantitative analysis of the naphthoquinone, plumbagin-a major constituent present in these species-using UPLC-UV. Microscopic examinations revealed the presence of distinctive differences in the anatomical features of the leaf, stem and root of the three species, and these can thus be used for identification and authentication of these species. UPLC-UV analysis showed the highest concentration of plumbagin in the roots of P. zeylanica (1.62% w/w) followed by the roots of P. indica (0.97% w/w) and then P. auriculata (0.33-0.53% w/w). In contrast, plumbagin was not detected in the stems and leaves of P. indica and in the leaves of P. auriculata, whereas very low concentrations (<0.02% w/w) of plumbagin were detected in the stems and leaves of P. zeylanica and in the stems of P. auriculata. HPTLC fingerprints of the leaf and root of the three species exhibited distinguishable profiles, while those of the stems were undifferentiated.

Larvicidal activity of extracts from three Plumbago spp against Anopheles gambiae.

Three Plumbago spp have been tested for mosquito larvicidal activity. The crude extracts exhibiting the highest larvicidal activity against Anopheles gambiae were hexane (LC50 = 6.4 microg/mL) and chloroform (LC50 = 6.7 microg/mL) extracts from Plumbago zeylanica Linn, chloroform (LC50 = 6.7 ug/mL) extract from Plumbago stenophylla Bull and ethyl acetate (LC50 = 4.1 microg/mL) extract from Plumbago dawei Rolfe. These LC50 values were within 95% confidence limits. 5-hydroxy-2-methyl-1,4-naphthoquinone (plumbagin) 1 (LC50 = 1.9 microg/mL) and beta-sitosterol 2 were characterised from ethyl acetate extract of root bark of P. dawei, a native medicinal plant growing in Kenya, based on spectral analysis and comparisons with data in literature.

Larvicidal activity of extracts from three Plumbago spp against Anopheles gambiae

Three Plumbago spp have been tested for mosquito larvicidal activity. The crude extracts exhibiting the highest larvicidal activity against Anopheles gambiae were hexane (LC50 = 6.4 μg/mL) and chloroform (LC50 = 6.7 μg/mL) extracts from Plumbago zeylanica Linn, chloroform (LC50 = 6.7 ug/mL) extract from Plumbago stenophylla Bull and ethyl acetate (LC50 = 4.1 μg/mL) extract from Plumbago dawei Rolfe. These LC50 values were within 95 percent confidence limits. 5-hydroxy-2-methyl-1,4-naphthoquinone (plumbagin) 1 (LC50 = 1.9 μg/mL) and β-sitosterol 2 were characterised from ethyl acetate extract of root bark of P. dawei, a native medicinal plant growing in Kenya, based on spectral analysis and comparisons with data in literature.

Intraspecific differentiation of Limonium wrightii (Plumbaginaceae) on northwestern Pacific Islands: rate heterogeneity in nuclear rDNA and its distance-independent geographic structure.

We conducted a molecular phylogenetic analysis of a coastal sea lavender in NW Pacific using ITS sequences of nrDNA. As a result, the ITS sequences of populations on the oceanic Daito Islands were considerably different from conspecific others in nine apomorphic substitutions. This finding indicates a distance-independent geographical structure of the genetic variation among islands in NW Pacific. The calibrated rate of nucleotide substitutions in the ITS regions within L. wrightii (average: 1.41x10(-8) substitutions per site per year) was higher than any other instances of substitution rates in flowering plants.

Historical and biological determinants of genetic diversity in the highly endemic triploid sea lavender Limonium dufourii (Plumbaginaceae).

Microsatellite markers were used to evaluate the genetic diversity and population genetic structure in the critically endangered Limonium dufourii (Plumbaginaceae), a highly endemic triploid species from the coasts of eastern Spain. Sixty-five alleles from 13 microsatellite regions were amplified in a sample of 122 individuals collected from the six extant populations. Microsatellite patterns were consistent with the triploid nature of L. dufourii. Alleles were unambiguously assigned to two different parental subgenomes in this hybrid species and the greater contribution of the diploid parental subgenome was confirmed. Eleven, 25 and 26 multilocus genotypes were recorded from the haploid, diploid and from the combined information of both subgenomes, respectively. Genetic diversity was mostly distributed among populations (72.06% of the total genetic variation). Genotypes from Marjal del Moro populations grouped into two highly structured clusters (88.41% of the total variance). The observed patterns of distribution of genetic diversity are interpreted to result from multiple hybridization events and isolation between populations. Threats to this species are mainly anthropogenic (urbanization and tourism pressure), although stochastic risks cannot be ignored. Therefore, in order to preserve extant genetic variation of L. dufourii, in situ strategies such as the preservation of its habitat are a high priority. Several recommendations in order to assist ex situ measures to guarantee the success of conservation strategies and maintain the relationships between individuals and populations are proposed.

Glacial-induced altitudinal migrations in Armeria (Plumbaginaceae) inferred from patterns of chloroplast DNA haplotype sharing.

In contrast to northern European areas where large-scale migrations occurred to recolonize territories after glacial periods, species in southern regions survived and diverged without large geographical displacements. As a result of the importance of orography in much of the southern areas, such displacements must have involved populations ascending or descending mountains. The present study provides support for glacial-induced altitudinal migrations from chloroplast phylogeographic patterns in Armeria (Plumbaginaceae) in southeast Spain. One hundred and five sequences of the trnL-F spacer were obtained from seven species. Fifteen different haplotypes were recognized, their genealogy was inferred, and associations with geography were explored using nested clade analysis. Seven instances were detected in which the same haplotype is shared by two or three species within a particular massif. In all the cases, at least one of the species involved displayed different haplotypes in other areas; in most, the haplotype shared is predominant either in one of the species involved or in the massif. These patterns of haplotype sharing strongly suggest horizontal transfer between species. In one of the massifs (Sierra Nevada) the three species involved in haplotype sharing (A. splendens, A. filicaulis ssp. nevadensis, A. villosa ssp. bernisii) occur at markedly different altitudinal belts. It is argued that altitudinal migrations within the contraction-expansion model provide the best explanation for the current pattern, and that at least in one case it resulted in the formation of a new hybrid taxon, A. filicaulis ssp. nevadensis.

Molecular evidence for the compilospecies model of reticulate evolution in Armeria (Plumbaginaceae).

Cladistic analyses of the nuclear ribosomal DNA (nrDNA) internal transcribed spacer (ITS) sequences from 55 samples corresponding to 34 taxa in the genus Armeria reveal that ITS sequence diversity among and within species utterly conflicts with patterns of morphological similarity. Three facts are apparent from the results here reported: (1) different samples of a single subspecies, A. villosa subsp. longiaristata, appear in three of the five major clades; (2) samples of at least one of the six subspecies of A. villosa appear in four of the five major clades; and (3) the composition of major clades shows greater congruence with the geographic origin of plants than with the traditional systematic arrangement based primarily on morphology. Specifically, the clades here termed Ia, II, III, and IV each encompass terminals restricted to geographically delimited areas. There are alternative explanations for the ITS pattern, but the most likely one is that nucleotide positions supporting the major clades are due, in some of the samples, to concerted evolution following horizontal transfer (gene flow) rather than to recency of common ancestry. This interpretation is consistent with previous systematic and experimental evidence and implies that reticulation in Armeria may be extensive. Harlan and de Wet (1963, Evolution 17:497-501) proposed the compilospecies concept to account for situations in which a genetically "aggressive" species captures portions of the genome of other sympatric species by means of extensive introgression. Evidence of extensive reticulation, ecological diversification, and geographic pattern indicates that A. villosa may fit the compilospecies concept, which is here supported on molecular grounds for the first time.