System level analysis of cacao seed ripening reveals a sequential interplay of primary and secondary metabolism leading to polyphenol accumulation and preparation of stress resistance.

Theobroma cacao and its popular product, chocolate, are attracting attention due to potential health benefits including antioxidative effects by polyphenols, anti-depressant effects by high serotonin levels, inhibition of platelet aggregation and prevention of obesity-dependent insulin resistance. The development of cacao seeds during fruit ripening is the most crucial process for the accumulation of these compounds. In this study, we analyzed the primary and the secondary metabolome as well as the proteome during Theobroma cacao cv. Forastero seed development by applying an integrative extraction protocol. The combination of multivariate statistics and mathematical modelling revealed a complex consecutive coordination of primary and secondary metabolism and corresponding pathways. Tricarboxylic acid (TCA) cycle and aromatic amino acid metabolism dominated during the early developmental stages (stages 1 and 2; cell division and expansion phase). This was accompanied with a significant shift of proteins from phenylpropanoid metabolism to flavonoid biosynthesis. At stage 3 (reserve accumulation phase), metabolism of sucrose switched from hydrolysis into raffinose synthesis. Lipids as well as proteins involved in lipid metabolism increased whereas amino acids and N-phenylpropenoyl amino acids decreased. Purine alkaloids, polyphenols, and raffinose as well as proteins involved in abiotic and biotic stress accumulated at stage 4 (maturation phase) endowing cacao seeds the characteristic astringent taste and resistance to stress. In summary, metabolic key points of cacao seed development comprise the sequential coordination of primary metabolites, phenylpropanoid, N-phenylpropenoyl amino acid, serotonin, lipid and polyphenol metabolism thereby covering the major compound classes involved in cacao aroma and health benefits.

Body size limits dim-light foraging activity in stingless bees (Apidae: Meliponini).

Stingless bees constitute a species-rich tribe of tropical and subtropical eusocial Apidae that act as important pollinators for flowering plants. Many foraging tasks rely on vision, e.g. spatial orientation and detection of food sources and nest entrances. Meliponini workers are usually small, which sets limits on eye morphology and thus quality of vision. Limitations are expected both on acuity, and thus on the ability to detect objects from a distance, as well as on sensitivity, and thus on the foraging time window at dusk and dawn. In this study, we determined light intensity thresholds for flight under dim light conditions in eight stingless bee species in relation to body size in a Neotropical lowland rainforest. Species varied in body size (0.8-1.7 mm thorax-width), and we found a strong negative correlation with light intensity thresholds (0.1-79 lx). Further, we measured eye size, ocelli diameter, ommatidia number, and facet diameter. All parameters significantly correlated with body size. A disproportionately low light intensity threshold in the minute Trigonisca pipioli, together with a large eye parameter P eye suggests specific adaptations to circumvent the optical constraints imposed by the small body size. We discuss the implications of body size in bees on foraging behavior.

Climatic and edaphic controls over tropical forest diversity and vegetation carbon storage.

Tropical rainforests harbor exceptionally high biodiversity and store large amounts of carbon in vegetation biomass. However, regional variation in plant species richness and vegetation carbon stock can be substantial, and may be related to the heterogeneity of topoedaphic properties. Therefore, aboveground vegetation carbon storage typically differs between geographic forest regions in association with the locally dominant plant functional group. A better understanding of the underlying factors controlling tropical forest diversity and vegetation carbon storage could be critical for predicting tropical carbon sink strength in response to projected climate change. Based on regionally replicated 1-ha forest inventory plots established in a region of high geomorphological heterogeneity we investigated how climatic and edaphic factors affect tropical forest diversity and vegetation carbon storage. Plant species richness (of all living stems >10 cm in diameter) ranged from 69 to 127 ha-1 and vegetation carbon storage ranged from 114 to 200 t ha-1. While plant species richness was controlled by climate and soil water availability, vegetation carbon storage was strongly related to wood density and soil phosphorus availability. Results suggest that local heterogeneity in resource availability and plant functional composition should be considered to improve projections of tropical forest ecosystem functioning under future scenarios.

Naphthalen-1-yl-(4-pentyloxynaphthalen-1-yl)methanone: a potent, orally bioavailable human CB1/CB2 dual agonist with antihyperalgesic properties and restricted central nervous system penetration.

Selective activation of peripheral cannabinoid CB1 receptors has the potential to become a valuable therapy for chronic pain conditions as long as central nervous system effects are attenuated. A new class of cannabinoid ligands was rationally designed from known aminoalkylindole agonists and showed good binding and functional activities at human CB1 and CB2 receptors. This has led to the discovery of a novel CB1/CB2 dual agonist, naphthalen-1-yl-(4-pentyloxynaphthalen-1-yl)methanone (13), which displays good oral bioavailability, potent antihyperalgesic activity in animal models, and limited brain penetration.

Two Unusual Methylidenecyclopropane Glucosides from Metaxya rostrata C.Presl.

Two new natural compounds, (1R,2E)-2-(6-hydroxyhexylidene)cyclopropyl-ß-D-glucopyranoside (1) and (6E)-6-[(2R)-2-(ß-D-glucopyranosyloxy)cyclopropylidene]hexanoic acid (2), glucosides of a very rare methylidenecyclopropane alcohol, as well as two known glycosides of phenolic acids, namely 4-O-ß-D-glucopyranosylcaffeic acid (3) and (E)-4-O-ß-D-glucopyranosylcoumaric acid (4), and methyl α-fructofuranoside (5) were isolated for the first time from the rhizomes of the tree fern Metaxya rostrata C.Presl. The structures were elucidated on the basis of detailed spectroscopic data analysis, and the structure of 1 was additionally confirmed by X-ray crystal-structure analysis.

Mechanical damage to pollen aids nutrient acquisition in Heliconius butterflies (Nymphalidae).

Neotropical Heliconius and Laparus butterflies actively collect pollen onto the proboscis and extract nutrients from it. This study investigates the impact of the processing behaviour on the condition of the pollen grains. Pollen samples (n = 72) were collected from proboscides of various Heliconius species and Laparus doris in surrounding habitats of the Tropical Research Station La Gamba (Costa Rica). Examination using a light microscope revealed that pollen loads contained 74.88 ± 53.67% of damaged Psychotria pollen, 72.04 ± 23.4% of damaged Psiguria/Gurania pollen, and 21.35 ± 14.5% of damaged Lantana pollen (numbers represent median ± first quartile). Damaged pollen grains showed deformed contours, inhomogeneous and/or leaking contents, or they were empty. Experiments with Heliconius and Laparus doris from a natural population in Costa Rica demonstrated that 200 min of pollen processing behaviour significantly increased the percentage of damaged pollen of Psychotria compared to pollen from anthers (P = 0.015, Z = -2.44, Mann-Whitney U-test). Examination of pollen loads from green house reared Heliconius butterflies resulted in significantly greater amounts of damaged Psiguria pollen after 200 min of processing behaviour compared to pollen from flowers (P < 0.001, Z = -4.583, Mann-Whitney U-test). These results indicate that pollen processing functions as extra oral digestion whereby pollen grains are ruptured to make the content available for ingestion.

Nitrogen nutrition during ontogeny of hemiepiphytic Clusia species.

This paper originates from a presentation at the IIIrd International Congress on Crassulacean Acid Metabolism, Cape Tribulation, Queensland, Australia, August 2001. We investigated the nitrogen (N) nutrition of Clusia osaensis, C. peninsulae and C. valerii during the seedling, epiphytic and hemiepiphytic phase in a lowland tropical forest in Costa Rica in order to elucidate nutritional adaptations of different plant growth stages to their habitat. Although all Clusia individuals were non-mycorrhizal, excised roots of seedlings, but also of epiphytic and hemiepiphytic stages, showed a distinct preference for glycine uptake. The shift in main rooting site from canopy soil to terrestrial soil was not reflected by changes in uptake rate or preference, although N availability and the composition of the available N pool changed significantly. High foliar N concentrations indicated that epiphytic seedlings seemed to be sufficiently supplied with N by maternal seed resources and canopy soils. With development, the epiphytic plants of Clusia may face N limitation due to higher N demands with increasing growth and restricted resources in the tree crowns. 15N natural abundance data indicate that epiphytes mainly accessed atmospheric and, to a lesser extent, canopy soil N sources and, after becoming terrestrially anchored, in the hemiepiphytic life stage exploited the larger nutrient reservoir of the ground soil. In consequence, Clusia species did not show an adaptation to the different N availability situations that they experienced whether canopy or ground-rooted.

Mode of photosynthesis during different life stages of hemiepiphytic Clusia species.

This paper originates from a presentation at the IIIrd International Congress on Crassulacean Acid Metabolism, Cape Tribulation, Queensland, Australia, August 2001. Carbon isotope fractionation and nocturnal acid accumulation in Clusia osaensis Hammel-ined., C. peninsulae Hammel-ined. and C. valerii Standl. were investigated during the seedling, epiphytic and hemiepiphytic phases in a lowland tropical forest in Costa Ricato study photosynthetic adaptations of different plant growth stages to their habitat. Foliar δ 13C values around -24 to -32‰ indicate predominant C3 fixation of CO2 and low crassulacean acid metabolism (CAM) activity in all three Clusia species. Only terrestrially rooted plants of C. osaensis showed increased CAM expression. However, all developmental stages exhibited significant CAM cycling as shown by significant day-night fluctuations of titratable protons and of malic and citric acid. In C. valerii and C. peninsulae, an increase in CAM expression with plant development was not observed, and CAM cycling in hemiepiphytic-stage plants was completely repressed during the high rainfall season. The expression of CAM in the three Clusia species is therefore not developmentally controlled but triggered by environmental factors such as water availability and light intensity. These factors remain relatively stable in this ecosystem and CAM is therefore not fully expressed. However, CAM cycling may be of ecophysiological significance in all life stages as it serves as a mechanism to improve carbon economy by reducing respiratory CO2 losses.