Rubiaceae Plant Research Articles

Anti-ulcer Effects of Paederia grevei (Rubiaceae) in Rat

Introduction: In Madagascar, the gastric ulcer prevalence raises to 2.3%. Generally, the gastric ulcer is one of the main problem that affects the digestive system that can be briefly defined as a loss of gastric substance wall reaching in-depth the muscular. In one hand, some medicinal plants are traditionally used against the gastric ulcer, among these plants is Paederia grevei (Rubiaceae plant family). This plant is known under the vernacular name of Lengomantsina. The goal of this work is to bring some highlight to the anti-ulcer activity of Paederia grevei and to validate its traditional use. Methods: This activity has been evaluated from three tests namely, mucoprotector, healing and anti-secretary. Results: The obtained results from experiences with rats show that Paederia grevei has a capacity mucoprotector by increasing the protection of the gastric mucous membrane in the rat after the administration of absolute ethanol. The protection of the gastric mucous membrane reached 79,02% at the batch treated with the hydroalcoolic amount of 600 mg/kg of the extract of Paederia grevei which is significantly meaningful compared to the pilot batch. After a continuous administration of indometacine, we observed that this extract improves the cicatrization of the lesions on the gastric wall. The surface of the lesions is of 8,33 ± 4,93 mm2 for the pilot batch whereas it is of 2,00 ± 1,69 for the treated batch with the amount of 600 mg/kg of Paederia grevei. In addition, the rough extract of Paederia grevei inhibits the acid gastric secretion, whose gastric pH contents increases by 2,09 ± 0,17 at the witness batches and 4,45 ± 0,69 at the batch treated with the amount of 600 mg/kg of the extract of Paederia grevei. Conclusion: One can thus say that our results confirm the traditional use of this plant.

Forest loss increases insect herbivory levels in human-altered landscapes

Insect herbivory has been observed to be affected by habitat loss and fragmentation, although the mechanisms by which these anthropogenic disturbances affect this process are not well understood. To aid in clarifying this issue, we assessed the relation between forest cover and leaf damage caused by herbivorous insects on a representative tropical forest understory plant family, the Rubiaceae. We measured leaf area loss of Rubiaceae plants in 20 forest sites located in the Brazilian Atlantic forest, and also tested whether variation in forest cover, abundance of insectivorous birds (predators) and of Rubiaceae plants (resources) could explain the observed variation in leaf damage. Herbivory levels varied between 2.6 and 12.5 percent leaf area lost and increased with decreasing forest cover, whereas the other explanatory variables did not provide additional explanatory power. Therefore, forest loss appears to be the main driver of changes in local herbivory, and ecological processes such as top-down and bottom-up control may not account for the deforestation-related increase in herbivory levels. Other mechanisms, for example leaf quality and/or the influence of the adjoining land uses, have to be explored in future studies.

Evidence of horizontal gene transfer between obligate leaf nodule symbionts.

Bacteria of the genus Burkholderia establish an obligate symbiosis with plant species of the Rubiaceae and Primulaceae families. The bacteria, housed within the leaves, are transmitted hereditarily and have not yet been cultured. We have sequenced and compared the genomes of eight bacterial leaf nodule symbionts of the Rubiaceae plant family. All of the genomes exhibit features consistent with genome erosion. Genes potentially involved in the biosynthesis of kirkamide, an insecticidal C7N aminocyclitol, are conserved in most Rubiaceae symbionts. However, some have partially lost the kirkamide pathway due to genome erosion and are unable to synthesize the compound. Kirkamide synthesis is therefore not responsible for the obligate nature of the symbiosis. More importantly, we find evidence of intra-clade horizontal gene transfer (HGT) events affecting genes of the secondary metabolism. This indicates that substantial gene flow can occur at the early stages following host restriction in leaf nodule symbioses. We propose that host-switching events and plasmid conjugative transfers could have promoted these HGTs. This genomic analysis of leaf nodule symbionts gives, for the first time, new insights in the genome evolution of obligate symbionts in their early stages of the association with plants.

Comprehensive approach for the detection of antifungal compounds using a susceptible strain of Candida albicans and confirmation of in vivo activity with the Galleria mellonella model

An efficient screening strategy for the identification of potentially interesting low-abundance antifungal natural products in crude extracts that combines both a sensitive bioautography assay and high performance liquid chromatography (HPLC) microfractionation was developed. This method relies on high performance thin layer chromatography (HPTLC) bioautography with a hypersusceptible engineered strain of Candida albicans (DSY2621) for bioactivity detection, followed by the evaluation of wild type strains in standard microdilution antifungal assays. Active extracts were microfractionated by HPLC in 96-well plates, and the fractions were subsequently submitted to the bioassay. This procedure enabled precise localisation of the antifungal compounds directly in the HPLC chromatograms of the crude extracts. HPLC-PDA-mass spectrometry (MS) data obtained in parallel to the HPLC antifungal profiles provided a first chemical screening about the bioactive constituents. Transposition of the HPLC analytical conditions to medium-pressure liquid chromatography (MPLC) allowed the efficient isolation of the active constituents in mg amounts for structure confirmation and more extensive characterisation of their biological activities. The antifungal properties of the isolated natural products were evaluated by their minimum inhibitory concentration (MIC) in a dilution assay against both wild type and engineered strains of C. albicans. The biological activity of the most promising agents was further evaluated in vitro by electron microscopy and in vivo in a Galleria mellonella model of C. albicans infection. The overall procedure represents a rational and comprehensive means of evaluating antifungal activity from various perspectives for the selection of initial hits that can be explored in more in-depth mode-of-action studies. This strategy is illustrated by the identification and bioactivity evaluation of a series of antifungal compounds from the methanolic extract of a Rubiaceae plant, Morinda tomentosa, which was used as a model in these studies.

UV/Visible spectra of a series of natural and synthesised anthraquinones: experimental and quantum chemical approaches.

Root decoctions of anthraquinone-containing plants are often taken as postpartum tonic and aphrodisiac. Anthraquinones are known for their diverse biological activities, especially antioxidant and anticancer. A series of 35 anthraquinones was generated by isolation from Rubiaceae plants and synthesis. Their UV/vis spectrum depends on the nature and relative positions of auxochromic substituents on the basic skeleton. To predict the maximum absorption bands for the current series of anthraquinones, excited sate calculations were performed using TD-DFT, CIS, ZINDO methods. The results showed that the use of PBE0 or its combination with B3LYP and B3P86 hybrid functionals are the most suitable to reproduce accurately the experimental λMAX. The structure–property relationships (SPRs) were established based on structural and electronic properties of the anthraquinones and showed (i) the importance of the number and position of OH groups and (ii) the positive contribution of the electrophilicity and hardness as electronic descriptors on position and amplitude of the maximum absorption bands.

Timarcha Latreille: A strange beetle and a living fossil

The genusTimarchaseems unique among Chrysomelidae: Chrysomelinae by having plesiomorphic characters such as genitalia with a ring-like tegmen, covered with a setose parameral cap, together with apomorphic characters, including apterism and fused elytra. The distribution of this genus is also very peculiar: circum Mediterranean and northwestern American. Food selection also seems rather stict, comprising 8 families and 20 genera of plants. Two of the subgenera (MetallotimarchaandAmericanotimarcha) share one plant family, the Ericaceae. The plant family Rubiaceae is also shared between three subgenera:Timarcha,Timarchostoma andMetallotimarcha. Reflex bleeding among diurnal species, aposematism, thanatosis, stomatic regurgitation are means of defense against predators, but not against parasites and commensals. A very archaic genus, probably originating in early Mesozoic, with around 100 taxa, and 50 subspecies. Mutations remain very common in the group and new species are possibly still emerging.

From ethnopharmacology to drug design

Nature’s diversity is one of the biggest resources of therapeutic lead compounds. Traditionally-used herbal remedies harbor a variety of bioactive compounds providing researchers with starting points for drug development. Ethnopharmacological investigations of uterotonic plant preparations identified a class of circular and disulfide-rich peptides, called cyclotides, to exhibit strong uterine contractions. In humans one of the physiological regulators of the myometrial contractility is the nonapeptide oxytocin acting on its cognate G protein-coupled receptors. They are considered to represent one of the most promising drug targets with ~30% of all currently marketed drugs acting on these transmembrane receptors. Based on observed similarities regarding the activity and structure of plant cyclotides with human oxytocin we analyzed the pharmacological principle of their action and identified the oxytocin and vasopressin 1a receptors as molecular targets of cyclotides from the Rubiaceae plant Oldenlandia affinis. Using a synthetic approach, the sequence of the native cyclotide kalata B7 was used to design oxytocin-like nonapeptides with nanomolar affinity and selectivity for the oxytocin receptor. This provides formal proof for the use of cyclotides as templates in peptide ligand design. At a more general level, mining of naturally-occurring peptides is a promising tool for the identification and the design of novel G protein-coupled receptor ligands.

Cyclotide discovery in Gentianales revisited--identification and characterization of cyclic cystine-knot peptides and their phylogenetic distribution in Rubiaceae plants.

Cyclotides are a unique class of ribosomally synthesized cysteine-rich miniproteins characterized by a head-to-tail cyclized backbone and three conserved disulfide-bonds in a knotted arrangement. Originally they were discovered in the coffee-family plant Oldenlandia affinis (Rubiaceae) and have since been identified in several species of the violet, cucurbit, pea, potato, and grass families. However, the identification of novel cyclotide-containing plant species still is a major challenge due to the lack of a rapid and accurate analytical workflow in particular for large sampling numbers. As a consequence, their phylogeny in the plant kingdom remains unclear. To gain further insight into the distribution and evolution of plant cyclotides, we analyzed ∼300 species of >40 different families, with special emphasis on plants from the order Gentianales. For this purpose, we have developed a refined screening methodology combining chemical analysis of plant extracts and bioinformatic analysis of transcript databases. Using mass spectrometry and transcriptome-mining, we identified nine novel cyclotide-containing species and their related cyclotide precursor genes in the tribe Palicoureeae. The characterization of novel peptide sequences underlines the high variability and plasticity of the cyclotide framework, and a comparison of novel precursor proteins from Carapichea ipecacuanha illustrated their typical cyclotide gene architectures. Phylogenetic analysis of their distribution within the Psychotria alliance revealed cyclotides to be restricted to Palicourea, Margaritopsis, Notopleura, Carapichea, Chassalia, and Geophila. In line with previous reports, our findings confirm cyclotides to be one of the largest peptide families within the plant kingdom and suggest that their total number may exceed tens of thousands.

Bioactive Metabolites from Endophytes Associated with Rubiaceous Plants from Uitm Puncak Alam Biological Reserve

Plant endophytes are an important and novel resource of natural bioactive compounds. In the past two decades, many valuable bioactive compounds with antimicrobial, cytotoxic and anticancer activities have been successfully discovered from the endophytic microorganisms. In this research, 54 endophytic fungi and 6 endophytic bacteria were isolated from leaves, stems, flowers and roots of the three plants species of Rubiaceae family collected in the biological reserve, UiTM Puncak Alam, Malaysia. The objective of this study is to isolate bioactive metabolites of endophytic microorganisms from Rubiaceae plants. The microorganisms were inoculated and fermented according to a standardized procedure. Each culture was extracted using ethyl acetate based on our standard operation procedure. The crude extracts were evaluated for preliminary screening of antimicrobial activity against Staphylococcus aureus ATCC 25923, Enterococcus Faecium ATCC 51585, Pseudomonas Aeruginosa ATCC 27853, Escherichia coli ATCC 25922 and Candida Albicans ATCC 44831, using the MTT method. Pleasingly, the antimicrobial effects of these crude extracts were slightly lower than standard antibiotics. The minimum inhibitory concentration (MIC) of the crude ethyl acetate extracts ranged from 0.1875 mg/mL to 1.5 mg/mL. The crude extracts were analyzed by high performance liquid chromatography (HPLC). HPLC fractionation followed by retesting of the fraction led to correlate chromatographic peaks with biological activity. Pure compound was subjected to standard spectrometry analyzes and its structure established. Data of compounds will be presented.

An Unprecedented Neolignan Skeleton from Chimarrhis turbinata

A lignan with a new skeleton named chimarrhinin (1) was isolated from an extract of the leaves of Chimarrhis turbinata, a Rubiaceae plant species. (13)C NMR spectrometric techniques including 1D and 2D experiments and HRESIMS provided unequivocal structural confirmation of this new C(6).C(3) skeleton type. The relative configuration of 1 was established by 2D (1)H-H analysis and J couplings, while its conformation was evaluated through molecular modeling using the RM1 semiempirical method, with the aid of coupling constants obtained by NMR analysis. The antioxidant activity of the new derivative 1 and two known and previously isolated phenolic derivatives (2 and 3) was investigated. An IC(50) value of 7.50 ± 0.5 μmol L(-1) was obtained for the new derivative 1, while 2 and 3 showed IC(50) values of 18.60 ± 0.4 and 18.50 ± 0.6 μmol, respectively.

Biochemical and biophysical characterization of a novel plant protein disulfide isomerase

We recently isolated a protein disulfide isomerase (PDI) from the Rubiaceae (coffee family) plant Oldenlandia affinis (OaPDI) and demonstrated that it facilitates the production of disulfide-knotted defense proteins called cyclotides. PDIs are major folding catalysts in the eukaryotic ER where they are responsible for formation, breakage, or shuffling of disulfide bonds in substrate polypeptides and are important chaperones in the secretory pathway. Here, we report the first detailed analysis of the oligomerization behavior of a plant PDI, based on characterization of OaPDI using various biochemical and biophysical techniques, including size-exclusion chromatography, NMR spectroscopy, surface plasmon resonance and atomic force microscopy. In solution at low concentration OaPDI comprises mainly monomers, but fractions of dimers and/or higher-order oligomers were observed at increased conditions, raising the possibility that dimerization and/or oligomerization could be a mechanism to adapt to the various-sized polypeptide substrates of PDI. Unlike mammalian PDIs, oligomerization of the plant PDI is not driven by the formation of intermolecular disulfide bonds, but by noncovalent interactions. The information derived in this study advances our understanding of the oligomerization behavior of OaPDI in particular but is potentially of broader interest for understanding the mechanism and role of oligomerization, and hence the catalytic and physiological mechanism, of the ubiquitous folding catalyst PDI.

Methanolic Extracts of Uncaria rhynchophylla Induce Cytotoxicity and Apoptosis in HT-29 Human Colon Carcinoma Cells

In this paper, we report the anticancer activities of Uncaria rhynchophylla extracts, a Rubiaceae plant native to China. Traditionally, Uncaria rhynchophylla has been used in the prevention and treatment of neurotoxicity. However, the cytotoxic activity of Uncaria rhynchophylla against human colon carcinoma cells has not, until now, been elucidated. We found that the methanolic extract of Uncaria rhynchophylla (URE) have cytotoxic effects on HT-29 cells. The URE showed highly cytotoxic effects via the MTT reduction assay, LDH release assay, and colony formation assay. As expected, URE inhibited the growth of HT-29 cells in a dose-dependent manner. In particular, the methanolic URE of the 500 microg/ml showed 15.8% inhibition against growth of HT-29 cells. It induced characteristic apoptotic effects in HT-29 cells, including chromatin condensation and sharking occurring 24 h when the cells were treated at a concentration of the 500 microg/ml. The activation of caspase-3 and the specific proteolytic cleavage of poly (ADP-ribose) polymerase were detected over the course of apoptosis induction. These results indicate that URE contains bioactive materials with strong activity, and is a potential chemotherapeutic agent candidate against HT-29 human colon carcinoma cells.

Antioxidant and Antimutagenic Activities of Randia echinocarpa Fruit

We report for the first time the antioxidant and antimutagenic activities of fractions from Randia echinocarpa fruit, which is a Rubiaceae plant native to Sinaloa, Mexico. This fruit has been traditionally used in the prevention or treatment of cancer, among other diseases. The pulp of the fruit was sequentially extracted with solvents of different polarity (i.e. hexane, chloroform, methanol and water). A high extraction yield was obtained with methanol (72.17% d.w.). The aqueous extract showed the highest content of phenolics (2.27 mg/g as ferulic acid equivalents) and the highest antioxidant activity based on the beta-carotene bleaching method (486.15). The commercial antioxidant BHT was used as control (835.05). Antimutagenic activity of the aqueous extract (0-500 microg/tube) was evaluated using the Salmonella microsuspension assay (YG1024 strain) and 1-NP as the mutagen (50 and 100 ng/tube). The aqueous extract was neither toxic nor mutagenic and the percentage of inhibition on 1-NP mutagenicity was 32 and 53% at doses of 50 and 100 ng/tube, respectively. The results of the double incubation assay suggest that the extract inhibited the mutagenicity of 1-NP by a combination of desmutagenic and bioantimutagenic effects.

Antibacterial, antifungal and cytotoxic activities of eight Asteraceae and two Rubiaceae plants from colombian biodiversity

Thirty crude extracts of eight plants belonging to Asteraceae and two to Rubiaceae families collected at different places from the Regional Natural Park Ucumari (RNPU), Colombia, were tested for their antibacterial activity against two Gram-positive and two Gram-negative bacteria and three fungi. Both the antibacterial and the antimycotic activities were tested by the agar well diffusion method. The cytotoxic activity on the same plant extracts was determined through the brine shrimp lethality bioassay. The extracts from the Asteraceae family were more bioactive against Bacillus subtilis and Staphylococcus aureus. The extracts of both families studied were bioactive against the fungi Candida albicans and Fusarium solani. In addition, the extracts of Asteraceae species displayed the greatest cytotoxic activities. However, the most important specie in this research was Gonzalagunia rosea Standl (Rubiaceae) because of the strong and moderate activities of the methanol and dichloromethane extracts against C. albicans and F. solani, respectively; as well as the strong cytotoxic activity of the methanol extract. None of these ten plants has previously been reported for their biological activities.

Optimization of microwave‐assisted extraction for alizarin and purpurin in Rubiaceae plants and its comparison with conventional extraction methods

A simple and rapid microwave-assisted extraction (MAE) procedure was developed and optimized for two common color pigments, alizarin and purpurin, in various samples of Rubiaceae plants. Several variables that can potentially affect the extraction efficiency, namely temperature, methanol concentration in the extractant mixture, time, and solvent volume were optimized by means of a central composite design approach. The results suggest that temperature and methanol concentration in the solvent mixture are statistically the most significant factors. The separation and quantitative determination of the pigments was carried out in less than 6 min by a developed high-performance liquid chromatographic method with UV detection at 250 nm. Under optimum operating conditions, MAE showed significantly higher recoveries than those obtained by the conventional extraction methods (ultrasonic and reflux extraction), ranging from 84 to 94%. In addition, a drastic reduction of the extraction time (20 min versus 6 h) and solvent consumption (20 versus 100 mL) was achieved with a reproducibility (RSDs < 10%) comparable with that provided by the reflux extraction as a reference method.

Variations in cyclotide expression in viola species.

Cyclotides, a family of approximately 50 mini-proteins isolated from various Violaceae and Rubiaceae plants, are characterized by their circular peptide backbone and six conserved cysteine residues arranged in a cystine knot motif. Cyclotides show a wide range of biological activities, making them interesting targets for both pharmaceutical and agrochemical research, but little is known about their natural function and the events that trigger their expression. An investigation of the geographical and seasonal variations of cyclotide profiles has been performed, using the native Australian violet, Viola hederacea, and the Swedish sweet violet, Viola odorata, as model plants. The results showed that in the Australian violet the relative peptide levels of some cyclotides remained almost constant throughout the year, while other cyclotides were present only at certain times of the year. Therefore, it appears that V. hederacea expresses a basic armory of cyclotides as well as special "add-ons" whose levels are influenced by external factors. In the Swedish violet, cyclotide levels were increased up to 14 times during the warmest period of the year. The larger variation in expression levels of the Swedish plants may be a reflection of a greater climatic variation.

Cloning of a cDNA encoding 1-deoxy- d-xylulose 5-phosphate synthase from Morinda citrifolia and analysis of its expression in relation to anthraquinone accumulation

In higher plants, there are two pathways leading to the formation of isoprenoids: the mevalonate pathway and the recently discovered methyl- d-erythritol 4-phosphate (MEP) pathway. It was shown that the isoprenoid moiety, constituting ring C of anthraquinones (AQs) in the plant family Rubiaceae, is formed from the MEP pathway. 1-Deoxy- d-xylulose 5-phosphate synthase (DXS), catalyzing the first step of the MEP pathway, is suggested to be a regulatory enzyme. In order to evaluate the potential regulatory role of the DXS in AQ biosynthesis, we identified a full-length cDNA encoding DXS from Morinda citrifolia, a member of the Rubiaceae family, by a PCR strategy. The cDNA encodes a protein of 722 amino acids with a calculated molecular mass of 78 kDa. The deduced amino acid sequence shares high homology with amino acid sequences of DXSs from other higher plants. Southern blot indicated the existence of a small family of DXS genes in M. citrifolia. Northern blot analysis showed that the DXS transcript levels were correlated with AQ accumulation, suggesting that AQ biosynthesis is regulated at the transcriptional level of the DXS gene.

More characters or more taxa for a robust phylogeny--case study from the coffee family (Rubiaceae).

Using different data sets mainly from the plant family Rubiaceae, but in parts also from the Apocynaceae, Asteraceae, Lardizabalaceae, Saxifragaceae, and Solanaceae, we have investigated the effect of number of characters, number of taxa, and kind of data on bootstrap values within phylogenetic trees. The percentage of supported nodes within a tree is positively correlated with the number of characters, and negatively correlated with the number of taxa. The morphological analyses are based on few characters and weakly supported trees are expected. The percentage of supported nodes is also dependent on the kind of data analyzed. In analyses of Rubiaceae based on the same number of characters, RFLP data give trees with higher percentage of supported nodes than rbcL and morphological data. We also discuss the support values for particular nodes at the familial and subfamilial levels. Two new data sets of ndhF and rbcL sequences of Rubiaceae are analyzed and together with earlier studies of the family we can conclude that the monophyly of the Rubiaceae is supported and within the family there are three well supported, but not easily characterized, large subfamilies, Rubioideae, Cinchonoideae s.s. and Ixoroideae s.l. There are also a few genera (Luculia and Coptosapelta) unclassified to subfamily.

ChemInform Abstract: Chemical Studies of Indole Alkaloids

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Chemical studies of indole alkaloids

Up to now, more than two thousands indole alkaloids having structural variety have been found from the three families of Gentianales: Loganiaceae, Apocynaceae, and Rubiaceae plants. These compounds generally possess characteristic biological activities and many of them are utilized for the medicinal purpose and for the lead-compounds to develop new synthetic drugs. The author of this review has been investigated the alkaloidal constituents of the above mentioned plants native to Japan and overseas countries. Especially, a cooperative work with Thai researchers concerning the plants in Thailand was developed. On the basis of biogenetic consideration of the structures of monoterpenoid indole alkaloids, chemical bond cleavage between the C3-N4 linkage in the C/D ring of indole alkaloids was discovered. It was speculated that, either in biogenesis, the flexibility of the ring-opening compounds having ten membered ring enabled the construction of indole alkaloids having highly strained polycyclic structures. Along with this conception, the syntheses of structurally complex indole alkaloids utilizing a biomimetic procedure have been investigated. In this review, recent results on the chemical synthesis of many skeletally varied Gelsemium alkaloids (Loganiaceae plant) are mainly described.