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Pollen flow in the distylous Palicourea fendleri (Rubiaceae): an experimental test of the Disassortative Pollen Flow Hypothesis.

The Disassortative Pollen Flow Hypothesis proposed by Darwin postulates that the relative position of anthers and stigmas in distylous flowers enhances pollen flow between flowers of different morphs (legitimate pollination), in comparison to flow between flowers of the same morph (illegitimate pollination). In order to test this hypothesis, we measured pollen transport, mediated by a trained Copper-rumped Hummingbird (Amazilia tobaci), between flowers of the distylous Palicourea fendleri under laboratory conditions. In individual tests, we offered to the hummingbird a pollen donor flower and two emasculated recipient flowers in a controlled sequence. After each foraging bout, we counted the number of pollen grains transported from the donor flower to the stigmas of both recipient flowers. In agreement with Darwin's hypothesis, we found that hummingbirds transport pollen of "pin" flowers in significantly higher numbers to legitimate "thrum" stigmas, even if previously visiting a "pin" flower. However, "thrum" pollen was deposited in greater numbers on illegitimate "thrum" stigmas. We interpret this asymmetry largely as the consequence of floral morphology; pollen flow was greater between anthers and stigmas that exhibit greater spatial matching. In P. fendleri, the position of floral organs along the corolla tube does not always precisely correspond. In our experimental system, the probability that the pollinator extracts a pollen grain from the anther and the probability of self-pollination were both dependent on the type of floral morph. We discuss the relevance of the latter findings in relation to other studies of pollen flow in heterostylous species.

Animals↗

Effects of partial throughfall exclusion on the phenology of Coussarea racemosa (Rubiaceae) in an east-central Amazon rainforest.

Severe droughts may alter the reproductive phenology of tropical tree species, but our understanding of these effects has been hampered by confounded variation in drought, light and other factors during natural drought events. We used a large-scale experimental reduction of throughfall in an eastern-central Amazon forest to study the phenological response to drought of an abundant subcanopy tree, Coussarea racemosa. We hypothesized that drought would alter the production and the timing of reproduction, as well as the number of viable fruits. The study system comprised two 1-ha plots in the Tapajos National Forest, Para, Brazil: a dry plot where 50% of incoming precipitation (80% throughfall) was diverted from the soil during the six-month wet season beginning in January 2000, and a wet plot that received natural rainfall inputs. Fruit production of C. racemosa was quantified every 15 days using 100 litter traps (0.5 m(2)) in each plot. The production of new leaves and flowers was recorded monthly for C. racemosa individuals. Soil water, pre-dawn leaf water potential and solar radiation were measured to help interpret phenological patterns. Over the approximately 3.5-year period (April 2000 through December 2003), total fruit production remained similar between plots, declining by 12%. In 2003, production was four times higher in both plots than in previous years. In the dry plot, fruit fall shifted 40 and 60 days later into the dry season in 2002 and 2003, respectively. Total fruit fall dry mass production was variable across the study period. Foliage and flower production coincided with peak irradiance early in the dry season until delays in flowering appeared in the dry plot in 2002 and 2003. Plant water stress, through its influence on leaf developmental processes and, perhaps, inhibition of photosynthesis, appears to have altered both the timing of fruit fall and the quality and number of seeds produced.

Brazil↗

Antimalarial activities of Tithonia diversifolia (Asteraceae) and Crossopteryx febrifuga (Rubiaceae) on mice in vivo.

Ethanolic extracts of the aerial part of Tithonia diversifolia and the stem bark of Crossopteryx febrifuga were investigated against early, residual (repository) and established malaria infections in vivo using Swiss albino mice at a dose range of 50-400 mg/kg per day. Chloroquine at 5 mg/kg per day was used as the positive control for the early and established infections while Pyrimethamine at 1.2 3/kg per day was used as the positive control for the residual infection test. Dose dependent chemo suppressive activities were obtained at the different levels of the infection tested. Tithonia diversifolia and Crossopteryx febrifuga gave some level of suppression of parasitaemia in the early and established infection stages. Tithonia diversifolia was active at 200 mg/kg per day in the repository test. The mean survival period of the mice treated with the extract in the established infection test was low, a possible indication of toxicity as a result of sub chronic administration of the extract. Crossopteryx febrifuga was inactive in the repository test. Beside the above limitations, the suppression of parasitaemia by the extracts at the highest dose was similar to chloroquine and pyrimathamine.

Administration, Oral↗

Antioxidant and 15-lipoxygenase inhibitory activities of the Malian medicinal plants Diospyros abyssinica (Hiern) F. White (Ebenaceae), Lannea velutina A. Rich (Anacardiaceae) and Crossopteryx febrifuga (Afzel) Benth. (Rubiaceae).

The African flora contains numerous medicinal plants whose biological and chemical properties are incompletely known. Antioxidant and radical scavenging properties of plants are subject to intensive research. In the work described here, we have investigated the antioxidant activity of the plants Diospyros abyssinica (root bark), Lannea velutina (root bark and stem bark) and Crossopteryx febrifuga (seeds). Extracts of different polarity were assayed for radical scavenging activity, using the stable free radical diphenylpicrylhydrazyl, and for inhibition of enzymatic lipid peroxidation mediated by soybean 15-lipoxygenase. All plants investigated showed activity, but there were large differences between plants and between extracts. In general, Diospyros abyssinica and Lannea velutina were richer in antioxidants than Crossopteryx febrifuga. Lipophilic extracts were not active as radical scavengers, but did inhibit 15-lipoxygenase. Semipolar extracts (80% aqueous ethanol and methanol) of Diospyros abyssinica and Lannea velutina showed the highest activity both as radical scavengers and lipoxygenase inhibitors, and also gave the highest extract yields. These plants therefore appear to be excellent sources of antioxidants.

Anacardiaceae↗

Antidiarrhoeal activity of the ethanol extract of Paederia foetida Linn. (Rubiaceae).

Paederia foetida L. is used as a remedy for diarrhoea and dysentery in Asia, but has not been investigated for its antidiarrhoeal properties. Antidiarrhoeal activity of 90% ethanol extract of Paederia foetida was investigated in this study using castor oil and magnesium sulphate-induced diarrhoea models in mice. The extract significantly increased the latent period of diarrhoea in both the models. In the castor oil study, the purging index (PI) value lowered in 1 h of the study at (100, 250 and 500 mg/kg) doses. The effect continued up to 6-h period only at 500 mg/kg dose. The plant notably reduced the purging index value in a dose-dependent manner in magnesium sulphate-induced diarrhoea. Paederia foetida, in general, reduced the gastrointestinal motility with barium sulphate milk both in 15- and 30-min time intervals. The extract significantly decreased the cisplatin-induced gastrointestinal motility at all doses at both time intervals. The extract also enhanced the morphine-induced reduction of motility at 500 mg/kg dose level at both time intervals. The results suggest that Paederia foetida showed antidiarrhoeal activity by inhibiting intestinal motility and justify its use in traditional medicine.

Animals↗

Antinociceptive activity of 3-O-beta-d-glucopyranosyl-23,24-dihydrocucurbitacin F from Hintonia standleyana (Rubiaceae).

Oral administration of a MeOH-CH(2)Cl(2) (1:1) extract of the stem bark of Hintonia standleyana (HSE) produced a dose-dependent antinociceptive effect when tested in mice using the writhing (150-750 mg/kg) and the hot-plate (150-600 mg/kg) models. From the active extract 3-O-beta-d-glucopyranosyl-23,24-dihydrocucurbitacin F (GDHCF), 5-O-[beta-d-apiofuranosyl-(1-->6)-beta-d-glucopyranosyl]-7-methoxy-3',4'-dihydroxy-4-phenylcoumarin (AG4-PC) and desoxycordifolinic acid (DCA) were isolated. GDHCF (10-100 mg/kg, p.o.) significantly reduced acetic acid-induced abdominal contortions and increased the hot-plate latency in comparison to vehicle-treated mice. Metamizol (50-100 mg/kg) and morphine (2.5-5 mg/kg) were used as positive controls, respectively. GDHCF-induced antinociception was partially blocked by naloxone (1 mg/kg, i.p.), l-NAME (150 mg/kg, i.p.) and glibenclamide (10mg/kg, i.p.) suggesting that its pharmacological effect could be due to the activation of the nitric oxide pathway, followed by the opening of the ATP-sensitive K(+) channels, as well as an activation of the opioid receptors.

Analgesics↗

Phenylpropanoid glucosides from leaves of Coussarea hydrangeifolia (Rubiaceae).

Phenylpropanoid glycosides, 1'-O-benzyl-alpha-L-rhamnopyranosyl-(1''-->6')-beta-D-glucopyranoside (1) and alpha-L-xylopyranosyl-(4''-->2')-(3-O-beta-D-glucopyranosyl)-1'-O-E-caffeoyl-beta-D-glucopyranoside (2), together with the known derivatives, 1,6-di-O-caffeoyl-beta-D-glucopyranoside (3), 1-O-(E)-caffeoyl-beta-D-glucopyranoside (4) and 1-O-(E)-feruloyl-beta-D-glucopyranoside (5), were isolated from leaves of Coussarea hydrangeifolia. Their structures were determined by IR, HRESIMS, and 1D and 2D NMR experiments, and their antioxidant activities, evaluated by assaying the free radical scavenging capacity using the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical as substrate. The antioxidant activities of 3 and 4 (IC50 values of 15.0 and 19.2 microM, respectively) were comparable to that of the standard positive control caffeic acid, whilst 2 and 5 were only weakly active and 1 was inactive.

Antioxidants↗

Iridoid glucosides from Randia spinosa (Rubiaceae).

An iridoid glucoside: randinoside, along with five known iridoids: galioside, deacetylasperulosidic acid methyl ester, scandoside methyl ester, geniposide and gardenoside, were isolated from the stems of Randia spinosa. The structures were determined by spectroscopic analysis, including 2D NMR techniques.

Iridoids↗

seco-iridoids from Calycophyllum spruceanum (Rubiaceae).

Three seco-iridoids 7-methoxydiderroside, 6'-O-acetyldiderroside and 8-O-tigloyldiderroside, were isolated from the wood bark of Calycophyllum spruceanum together with the known iridoids loganetin, loganin and the seco-iridoids secoxyloganin, kingiside and diderroside. Their structures were elucidated by means of NMR and MS spectral data analysis. Using NOE correlations and coupling constants, the relative stereochemistry of the new derivatives was established. 7-Methoxydiderroside, 6'-O-acetyldiderroside and the known secoxyloganin and diderroside showed in vitro activity against trypomastigote forms of Trypanosoma cruzi, with IC(50) values of 59.0, 90.2, 74,2 and 84.9 microg/mL, respectively and were compared to the standard gentian violet (IC(50) 7.5 microg/ml).

Animals↗

Iridoid and seco-iridoid glucosides from Chioccoca alba (Rubiaceae).

Phytochemical investigation of Chioccoca alba afforded three new iridoids, alboside I, alboside II and alboside III, and a new seco-iridoid alboside V. Alboside IV showed moderate activity towards the DNA repair-deficient mutant RS321 of Saccharomyces cerevisiae. The structural elucidation of the new compounds was performed by ES-MS and by 1D and 2D NMR spectroscopic methods.

Glucosides↗

Heteromorphic incompatibility and efficiency of pollination in two distylous Pentanisia species (Rubiaceae).

BACKGROUND AND AIMS: Distyly has been hypothesized to promote cross-pollination by reducing intrafloral and geitonogamous self-pollination, and enhancing intermorph pollination. Distylous plants typically display both reciprocal herkogamy and a heteromorphic incompatibility system, which allows mating only between morphs. Distyly and its pollination consequences were examined in two Pentanisia species with long-tubed flowers which are pollinated almost exclusively by butterflies. METHODS: Anther and stigma heights were measured to quantify reciprocal herkogamy. The type of incompatibility system was determined by observing pollen tubes and seed production following controlled hand pollination. Pollen loads on pollinators and stigmas were also examined to assess the efficiency of intermorph pollen flow. KEY RESULTS: Pentanisia prunelloides and P. angustifolia exhibit reciprocal herkogamy and a host of ancillary dimorphisms, including pollen colour, exine sculpturing, stigmatic papilla shape and floral-tube pubescence. Controlled hand-pollinations revealed the presence of a strong heteromorphic incompatibility system in both species. The site of incompatibility differed between the morphs; intramorph pollen tubes were blocked in the style of the short-styled morph and on the stigmatic surface of the long-styled morph. Butterflies carried pollen from the short- and long-styled morphs primarily on their head and proboscis, respectively. Natural pollination resulted in a higher proportion of pollen transfer from long- to short-styled plants than vice versa. Nevertheless, fruit set did not differ between morphs. CONCLUSIONS: Both Pentanisia species are fully distylous. Reciprocal herkogamy results in pollen from the two morphs being carried on different locations on pollinators' bodies, which in turn promotes intermorph pollination. Intramorph pollination does not result in fertilization, because of an effective heteromorphic incompatibility system.

Flowers↗

Elaborate petals in Australian Spermacoce (Rubiaceae) species: morphology, ontogeny and function.

BACKGROUND AND AIMS: Australian Spermacoce species display various types of elaborate petals. Their precise morphology, ontogenetic origin, and function are hitherto unknown. The aim of the present paper is to unravel the development and nature of the diverse types of elaborate petals in Spermacoce through a floral ontogenetic study. METHODS: The floral ontogeny of six species characterized by different types of corolla appendages was studied by scanning electron microscopy and light microscopy. In order to elucidate the possible functions of the elaborate petals, field observations were conducted as well. KEY RESULTS: Scanning electronmicrographs show that full-grown petals of Spermacoce lignosa, S. phaeosperma and S. redacta bear appendages on their ventral side. Despite their different appearance at anthesis, the appendages develop very similarly in all three species. They are initiated at the same developmental stage and are first visible as two arcs of primordia converging from the upper margins of the petal towards its midvein and downwards. In S. brevidens, S. caudata and S. erectiloba, the full-grown petals have two long, concave protuberances, which develop from the tissue at both sides of the petal's mid-vein. In these three species, initiation of appendages on the ventral side of the petals is also observed, but they are hardly visible on the mature petals. The two types of elaborate petals tightly enclose the anthers, both in bud and during most of the flowering period. CONCLUSIONS: Among Australian Spermacoce species, two types of elaborate petals can be distinguished. The former hypothesis that the two types of elaborate petals are essentially homologous is here rejected. Field investigations point out that the elaborate petals might play a role in the pollination biology of the species.

Australia↗

Nucleotide sequence diversity at the methionine synthase locus in endangered Dunnia sinensis (Rubiaceae): an evaluation of the positive selection hypothesis.

Methionine synthase is a key enzyme for the synthesis of the aspartate-derived methionine, the immediate precursor of S-adenosyl-methionine, which has been illustrated to be associated with plant growth and pathogen interactions. In this study we tested the positive selection hypothesis of molecular evolution of the methionine synthase gene in Dunnia sinensis. In the entire sample of 87 sequences, 22 haplotypes of introns and 16 haplotypes of exons were identified. An excess of polymorphism over the neutral expectation for the class of unique nucleotide polymorphisms was observed in both exon and intron sequences. Ten replacement substitutions versus six synonymous substitutions among lineages, although nonsignificant, revealed that some advantageous mutants might have been favored. The distribution of d(N)/d(S) > 1 at nodes between closely related haplotypes in the gene network also indicated weak and variable positive selection. Nevertheless, low levels of genetic diversity in exons (theta; = 0.0052) and introns (theta; = 0.0070) of the methionine synthase gene of the outcrossing Dunnia were also attributed to the endangered status of the species. The atpB-rbcL intergenic spacer of cpDNA and the ribosomal internal transcribed spacer of mtDNA were used to discern the relative effectiveness of natural selection from intrinsic evolutionary forces. The low levels of nucleotide polymorphisms in both organelle spacers and the significant population differentiation reflected the effect of population-species history and demography. Two major lineages of the methionine synthase gene genealogy were recovered corresponding to two geographic regions, a result that was consistent with organelle phylogenies. Both past fragmentation and recent habitat disturbance causing complete bottlenecks may have resulted in population decline and geographic isolation and may have led to the depletion of genetic variation at loci in nuclear and organelle genomes.

5-Methyltetrahydrofolate-Homocysteine S-Methyltran↗

Variation among populations of Diodia teres (Rubiaceae) in environmental maternal effects.

Previous studies have quantified variation in environmental maternal effects (EME) within populations, but these effects could differ among populations as well. In this study we grew clonal replicates of individuals from three populations of the annual plant Diodia teres in their native and non-native environments. Our goal was to estimate the effects of maternal environment and maternal population on seed and seedling traits. Seeds that were produced in this field study were then planted in two soil types to quantify effects of the offspring environment on seedling traits. There was substantial variation among populations for seed weight. We found population variation for EME, and maternal environment by offspring environment interactions. We conclude that variation among populations in EME may be an unrecognized component of local adaptation, and that attempts to control maternal effects by statistically accounting for variation in seed weight may be ineffective.

Adaptation, Physiological↗

Hepatoprotective properties in the rat of Mitracarpus scaber (Rubiaceae).

The effect of Mitracarpus scaber on carbon tetrachloride-induced acute liver damage in the rat has been evaluated. Results showed that treatment with Mitracarpus scaber decoction resulted in significant hepatoprotection against CCl4-induced liver injury both in-vivo and in-vitro. In-vivo, Mitracarpus scaber pretreatment reduced levels of serum glutamate-oxalate-transaminase (P < 0.01 for 250, 500 and 1000 mg kg(-1)) and serum glutamate-pyruvate-transaminase (P < 0.05 for 250 mg kg(-1) and P < 0.01 for 1000 mg kg(-1)) previously increased by administration of CCl4. In-vitro results indicated that addition to the culture medium of Mitracarpus scaber extracts significantly reduced glutamate-oxalate-transaminase (P < 0.05 for 100 microg mL(-1) and P < 0.01 for 10 and 1000 microg mL(-1)) and lactate dehydrogenase activity (P < 0.05 for 10 microg mL(-1)). Mitracarpus treatment also resulted in a good ( > 93%) survival rate for the CCl4-intoxicated hepatocytes as demonstrated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Moreover, as in the in-vitro assay, Mitracarpus scaber had radical-scavenging properties, shown by its reaction with the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical (EC50, the extract concentration resulting in a 50% reduction in the absorbance of DPPH blank solution, = 41.64+/-1.5 microg mL(-1)). The results of this study showed that Mitracarpus scaber had antihepatotoxic potential, a finding which supports the validity of traditional usage of this drug in Mali for the treatment of liver diseases.

Animals↗

Iridoids and anthraquinones from the Malaysian medicinal plant, Saprosma scortechinii (Rubiaceae).

A further investigation of the leaves and stems of Saprosma scortechinii afforded 13 compounds, of which 10 are new compounds. These were elucidated as the bis-iridoid glucosides, saprosmosides G (1) and H (2), the iridoid glucoside, 6-O-epi-acetylscandoside (3), and the anthraquinones, 1-methoxy-3-hydroxy-2-carbomethoxy-9,10-anthraquinone (4), 1-methoxy-3-hydroxy-2-carbomethoxy-9,10-anthraquinone 3-O-beta-primeveroside (5), 1,3-dihydroxy-2-carbomethoxy-9,10-anthraquinone 3-O-beta-primeveroside (6), 1,3,6-trihydroxy-2-methoxymethyl-9,10-anthraquinone (7), 1-methoxy-3,6-dihydroxy-2-hydroxymethyl-9,10-anthraquinone (8), 1,3,6-trihydroxy-2-hydroxymethyl-9,10-anthraquinone 3-O-beta-primeveroside (9), and 3,6-dihydroxy-2-hydroxymethyl-9,10-anthraquinone (10). Structure assignments for all compounds were established by means of mass and NMR spectroscopies, chemical methods, and comparison with published data. The new anthraquinones were derivatives of munjistin and lucidin.

Anthraquinones↗