Secondary Metabolites In Fruits Research Articles

The roles of non-structural carbohydrates in fruiting: a review focusing on mango (Mangifera indica).

Reproductive development of fruiting trees, including mango (Mangifera indica L.), is limited by non-structural carbohydrates. Competition for sugars increases with cropping, and consequently, vegetative growth and replenishment of starch reserves may reduce with high yields, resulting in interannual production variability. While the effect of crop load on photosynthesis and the distribution of starch within the mango tree has been studied, the contribution of starch and sugars to different phases of reproductive development requires attention. This review focuses on mango and examines the roles of non-structural carbohydrates in fruiting trees to clarify the repercussions of crop load on reproductive development. Starch buffers the plant's carbon availability to regulate supply with demand, while sugars provide a direct resource for carbon translocation. Sugar signalling and interactions with phytohormones play a crucial role in flowering, fruit set, growth, ripening and retention, as well as regulating starch, sugar and secondary metabolites in fruit. The balance between the leaf and fruit biomass affects the availability and contributions of starch and sugars to fruiting. Crop load impacts photosynthesis and interactions between sources and sinks. As a result, the onset and rate of reproductive processes are affected, with repercussions for fruit size, composition, and the inter-annual bearing pattern.

Transcriptome analysis reveals important regulatory factors for condensed tannins synthesis in acorn.

Condensed tannins are widely present in the fruits and seeds of plants and effectively prevent them from being eaten by animals before maturity due to their astringent taste. In addition, condensed tannins are a natural compound with strong antioxidant properties and significant antibacterial effects. Four samples of mature and near-mature Quercus fabri acorns, with the highest and lowest condensed tannin content, were used for genome-based transcriptome sequencing. The KEGG enrichment analysis revealed that the differentially expressed genes (DEGs) were highly enriched in phenylpropanoid biosynthesis and starch and sucrose metabolism. Given that the phenylpropanoid biosynthesis pathway is a crucial step in the synthesis of condensed tannins, we screened for significantly differentially expressed transcription factors and structural genes from the transcriptome data of this pathway and found that the expression levels of four MADS-box, PAL, and 4CL genes were significantly increased in acorns with high condensed tannin content. The quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) experiment further validated this result. In addition, yeast one-hybrid assay confirmed that three MADS-box transcription factors could bind the promoter of the 4CL gene, thereby regulating gene expression levels. This study utilized transcriptome sequencing to discover new important regulatory factors that can regulate the synthesis of acorn condensed tannins, providing new evidence for MADS-box transcription factors to regulate the synthesis of secondary metabolites in fruits.

Metabolome, Plant Hormone, and Transcriptome Analyses Reveal the Mechanism of Spatial Accumulation Pattern of Anthocyanins in Peach Flesh.

Anthocyanins are important secondary metabolites in fruits, and anthocyanin accumulation in the flesh of peach exhibits a spatial pattern, but the relevant mechanism is still unknown. In this study, the yellow-fleshed peach, cv. 'Jinxiu', with anthocyanin accumulation in the mesocarp around the stone was used as the experimental material. Red flesh (RF) and yellow flesh (YF) were sampled separately for flavonoid metabolite (mainly anthocyanins), plant hormone, and transcriptome analyses. The results showed that the red coloration in the mesocarp was due to the accumulation of cyanidin-3-O-glucoside, with an up-regulation of anthocyanin biosynthetic genes (F3H, F3'H, DFR, and ANS), transportation gene GST, and regulatory genes (MYB10.1 and bHLH3). Eleven ERFs, nine WRKYs, and eight NACs were also defined as the candidate regulators of anthocyanin biosynthesis in peach via RNA-seq. Auxin, cytokinin, abscisic acid (ABA), salicylic acid (SA), and 1-aminocyclopropane-1-carboxylic acid (ACC, ethylene precursor) were enriched in the peach flesh, with auxin, cytokinin, ACC, and SA being highly accumulated in the RF, but ABA was mainly distributed in the YF. The activators and repressors in the auxin and cytokinin signaling transduction pathways were mostly up-regulated and down-regulated, respectively. Our results provide new insights into the regulation of spatial accumulation pattern of anthocyanins in peach flesh.

Comprehensive Characterization of Secondary Metabolites in Fruits and Leaves of Cloudberry (Rubus chamaemorus L.)

Cloudberry (Rubus chamaemorus L.) is a circumpolar boreal plant rich in bioactive compounds and is widely used in food and in folk medicine. In this study, a combination of two-dimensional NMR spectroscopy and liquid chromatography–high-resolution mass spectrometry was used for the comprehensive characterization of secondary metabolites in cloudberry lipophilic and hydrophilic extracts. Special attention was paid to the leaf extractives, which are highly enriched in polyphenolic compounds, the content of which reaches 19% in the extract (in gallic acid equivalent). The chemical composition of the polyphenolic fraction is represented mainly by the glycosylated derivatives of flavonoids, hydroxycinnamic (primarily caffeic), gallic (including the structure of galloyl ascorbate) and ellagic acids, catechin, and procyanidins. The contents of aglycones in the polyphenolic fraction were 64 and 100 mg g−1 for flavonoids and hydroxycinnamic acids, respectively, while the content of free caffeic acid was 1.2 mg g−1. This determines the exceptionally high antioxidant activity of this fraction (750 mg g−1 in gallic acid equivalent) and the ability to scavenge superoxide anion radicals, which is 60% higher than that of Trolox. The lower polar fractions consist mainly of glycolipids, which include polyunsaturated linolenic acid (18:3), pentacyclic triterpenic acids, carotenoid lutein, and chlorophyll derivatives, among which pheophytin a dominates. Along with the availability, the high antioxidant and biological activities of cloudberry leaf extracts make them a promising source of food additives, cosmetics, and pharmaceuticals.

Influence of Living Mulch and Nitrogen Dose on Yield and Fruit Quality Parameters of Malus domestica Borkh. cv. ‘Sampion’

This study was conducted to estimate the yield, and to identify and quantify primary and secondary metabolites in fruit of Malus domestica Borkh. cv. ‘Sampion’ under two agrotechnical factors: the floor management (herbicide fallow and living mulch) and the dose of nitrogen (50, 80, 110, and 140 kg ha−1). Compared to herbicide fallow, living mulch did not decrease yield. Research showed a rich composition of phenolic and volatile organic compounds in apples, which varied with the evaluated factors, as well as with the weather conditions during the vegetation season. The precipitation deficit and high summer temperatures did not contribute to proper fruit growth and development and led to a higher content of phenolic compounds in the fruit flesh from trees in herbicide fallow compared to living mulch. Living mulch, which could be a factor regulating the availability of nitrogen to trees, stimulated the synthesis of anthocyanins, which was also potentiated by low average temperatures at harvest time, resulting in a large area of fruit skin red blush.

Resolving the developmental distribution patterns of polyphenols and related primary metabolites in bilberry (Vaccinium myrtillus) fruit

Bilberry (Vaccinium myrtillus) is a commercially important wild berry species, which accumulates high amounts of polyphenols, particularly anthocyanins, in the skin and flesh. Whilst a number of studies have quantified these phytochemicals in intact ripe bilberry fruit, we extend the current knowledge by investigating the spatial distribution of anthocyanin-associated polyphenols in fruit tissue, and study their links with primary metabolism during ripening. To address this, we used LC-MS and mass spectrometry imaging to measure and map primary and secondary metabolites in fruit. Correlation analysis showed that five sugars displayed strong positive correlations with anthocyanin accumulation, whereas all amino acids were negatively correlated. The accumulation patterns of polyphenols correlated in fruit skin and flesh, but altered with development. Finally, spatial segmentation analysis revealed that the chemical signatures of ripening first appear at defined regions under the skin and rapidly expand to encompass the entire fruit at the eating-ripe stage.

Fruits, frugivores, and the evolution of phytochemical diversity

Plants produce an enormous diversity of secondary metabolites, but the evolutionary mechanisms that maintain this diversity are still unclear. The interaction diversity hypothesis suggests that complex chemical phenotypes are maintained because different metabolites benefit plants in different pairwise interactions with a diversity of other organisms. In this synthesis, we extend the interaction diversity hypothesis to consider that fruits, as potential hotspots of interactions with both antagonists and mutualists, are likely important incubators of phytochemical diversity. We provide a case study focused on the Neotropical shrub Piper reticulatum that demonstrates: 1) secondary metabolites in fruits have complex and cascading effects for shaping the outcome of both mutualistic and antagonistic fruit–frugivore interactions, and; 2) fruits can harbor substantially higher levels of phytochemical diversity than leaves, even though leaves have been the primary focus of plant chemical ecology research for decades. We then suggest a number of research priorities for integrating chemical ecology with fruit–frugivore interaction research and make specific, testable predictions for patterns that should emerge if fruit interaction diversity has helped shape phytochemical diversity. Testing these predictions in a range of systems will provide new insight into the mechanisms driving frugivory and seed dispersal and shape an improved, whole‐plant perspective on plant chemical trait evolution.

Metabolomic profile combined with transcriptomic analysis reveals the value of UV-C in improving the utilization of waste grape berries

Table grape is a popular fruit worldwide. The quality of the appearance of table grapes directly affects their commercial value. Table grape bunches are usually carefully managed during production. At different developmental stages, a large number of grape berries are pruned as waste for commercial appearance, which leads to wasted resources. Ultraviolet-C (UV-C) can regulate the accumulation of secondary metabolites in fruits. In this study, metabolomic profile was combined with transcriptomic analysis technology to explore the value of UV-C in improving the utilization of waste grapes. The berries of the 'Jumeigui' grape were subjected to UV-C radiation treatment in the green-berry stage, veraison stage, and maturation stage. The results showed that UV-C could brown grape berries and decrease their sugar content at different developmental stages. Compared with other samples, those treated with UV-C in the veraison stage had the most upregulated metabolites, while samples in the green-berry stage had the most down-regulated metabolites. UV-C promoted the accumulation of stilbenes and some flavonoids in the berries at each developmental stage (especially at the green-berry and veraison stages). Compared with other stages, UV-C treatment during the veraison stage led to the highest number of upregulated genes related to transcription factors, protein modification, indole-3-acetic acid (IAA), abscisic acid (ABA), gibberellin acid (GA), receptor kinases, and Ascorbic acid/Glutathione (Ascorb/Gluath). UV-C might promote the accumulation of phenolic components by upregulating the expression of their biosynthesis related genes. UV-C may be an effective in vitro approach for improving the application value of waste grape berries by enhancing the accumulation of the nutritious phenolic components.

Flavonoids and Their Anti-Diabetic Effects: Cellular Mechanisms and Effects to Improve Blood Sugar Levels.

Diabetes mellitus (DM) is a prevailing global health metabolic disorder, with an alarming incidence rate and a huge burden on health care providers. DM is characterized by the elevation of blood glucose due either to a defect in insulin synthesis, secretion, binding to receptor, or an increase of insulin resistance. The internal and external factors such as obesity, urbanizations, and genetic mutations could increase the risk of developing DM. Flavonoids are phenolic compounds existing as secondary metabolites in fruits and vegetables as well as fungi. Their structure consists of 15 carbon skeletons and two aromatic rings (A and B) connected by three carbon chains. Flavonoids are furtherly classified into 6 subclasses: flavonols, flavones, flavanones, isoflavones, flavanols, and anthocyanidins. Naturally occurring flavonoids possess anti-diabetic effects. As in vitro and animal model’s studies demonstrate, they have the ability to prevent diabetes and its complications. The aim of this review is to summarize the current knowledge addressing the antidiabetic effects of dietary flavonoids and their underlying molecular mechanisms on selected pathways: Glucose transporter, hepatic enzymes, tyrosine kinase inhibitor, AMPK, PPAR, and NF-κB. Flavonoids improve the pathogenesis of diabetes and its complications through the regulation of glucose metabolism, hepatic enzymes activities, and a lipid profile. Most studies illustrate a positive role of specific dietary flavonoids on diabetes, but the mechanisms of action and the side effects need more clarification. Overall, more research is needed to provide a better understanding of the mechanisms of diabetes treatment using flavonoids.

Dynamics of soluble sugars and secondary metabolites in fruit of Juglans australis attacked by Anastrepha fraterculus and Ceratitis capitata (Diptera: Tephritidae)

The development and fitness of phytophagous insects are tightly linked to the nutritional quality of their host plants and many studies have examined the influence of primary and secondary metabolites of plants and their effects on the development of insects. Herbivore tactics to modify plant metabolic pathways to lower host toxicity need to be better understood as they are critical to a better understanding of herbivore–host plant relationships. To contribute to this end, in this study we analyzed temporal patterns of glucose, sucrose, fructose, and total soluble sugar contents, as well as tannins, phenols, and flavonoids in the mesocarp of fruit of native walnut (Juglans australis), uninfested and infested by Anastrepha fraterculus and Ceratitis capitata (sometimes simultaneously in a single fruit). Both fly species are polyphagous tephritids whose larvae feed on a wide variety of hosts. We observed a high correlation between infestation and adult emergence of these two insects which was positive in the case of sugar content and negative in the case of toxic secondary metabolites in fruit. At particular ripening stages, infested fruit contained higher levels of sugars and lower levels of phenols and tannins than non-infested fruit. We discuss the possibility that A. fraterculus and C. capitata, each with different egg-laying strategies, may modify metabolical pathways in the fruit for their own benefit through larval activity with the help of bacteria in their gut. Alternatively, the patterns observed may be simple side effects of larval feeding and associated growth of opportunistic microorganisms.

Human exposure modelling of quercetin in onions (Allium cepa L.) following thermal processing

Post-harvest treatment can influence levels of secondary metabolites in fruits and vegetables. Onions contain high levels of quercetin but are commonly heat-treated before consumption. Hence, the objective of this study was to examine the effect of cooking treatments on the flavonoid (3,4′-Qdg and 4′-Qmg) concentrations in onion and to determine, by simulation modelling, probable human exposure. Onion samples (n=3) were cooked using three processes (fry, bake and steam) for three time intervals (5, 10 and 15min). Frying (<10min) was the ideal cooking method which retained concentrations of 3,4′-Qdg and 4′-Qmg at >50%. Thermal processing (>10min) was shown to decrease quercetin content in all samples. The simulation model predicted human absorption and exposure. Steaming (15min) resulted in the lowest quercetin exposure, with mean values of 4000 and 400μg/day for 3,4′-Qdg and 4′-Qmg, respectively. Untreated onions had mean exposures of 14,000 and 3000μg/day for 3,4′-Qdg and 4′-Qmg, respectively.

Chemical ecology of fruit defence: synergistic and antagonistic interactions among amides from Piper

Summary Although ripe, fleshy fruits function primarily to attract seed dispersers, they must also be defended against diverse communities of seed predators and pathogens. For some plants, the concentration and diversity of secondary metabolites in fruits can exceed that of leaves and other plant parts, but little is known about the functional significance of the suites of compounds found in fruits. Fruit secondary metabolites may function in defence, or they may play a variety of other roles in seed development and dispersal. In this study, we conducted a series of experiments to test the effects of amides, a highly diverse class of secondary metabolites found in Piper fruits, on a variety of antagonistic fruit pests, including an insect seed predator (Sibaria englemani, Pentatomidae) and three unidentified species of fungi isolated from ripe Piper reticulatum (Piperaceae) fruits. We tested the effects of amide‐rich extracts from unripe and ripe fruits of P. reticulatum and the effects of two pure compounds, piperine and piplartine, presented alone and in combination. Amide‐rich extracts from unripe and ripe fruits had no effect on insect feeding preferences, but strong negative effects on fungal growth rates. A comparison of the relative bioactivity of unripe and ripe fruit extracts, controlling for concentration, showed that the specific composition of compounds in unripe fruits provides a more effective defence than that of ripe fruits against two of the three fungal species tested. Pure amides had variable effects on insect feeding preferences and strong negative effects on fungal growth rates. Tests of the bioactivity of two pure amides, presented alone and in combination, showed that the same two compounds can interact either synergistically or antagonistically in mixtures depending on the particular consumer involved. Together, these results suggest that the secondary metabolites in fruits may be a key characteristic contributing to fruit defence and plant reproductive success. Specifically, our results emphasize: (i) the potential for slight changes in the composition of mixtures to alter the efficacy of defence; and (ii) the potential for complex interactions among compounds in mixtures that can alter the bioactivity of secondary metabolites differentially among different consumers.

The Distribution of Fruit and Seed Toxicity during Development for Eleven Neotropical Trees and Vines in Central Panama

Secondary compounds in fruit mediate interactions with natural enemies and seed dispersers, influencing plant survival and species distributions. The functions of secondary metabolites in plant defenses have been well-studied in green tissues, but not in reproductive structures of plants. In this study, the distribution of toxicity within plants was quantified and its influence on seed survival was determined in Central Panama. To investigate patterns of allocation to chemical defenses and shifts in allocation with fruit development, I quantified variation in toxicity between immature and mature fruit and between the seed and pericarp for eleven species. Toxicity of seed and pericarp was compared to leaf toxicity for five species. Toxicity was measured as reduced hyphal growth of two fungal pathogens, Phoma sp. and Fusarium sp., and reduced survivorship of brine shrimp, Artemia franciscana, across a range of concentrations of crude extract. I used these measures of potential toxicity against generalist natural enemies to examine the effect of fruit toxicity on reductions of fruit development and seed survival by vertebrates, invertebrates, and pathogens measured for seven species in a natural enemy removal experiment. The seed or pericarp of all vertebrate- and wind-dispersed species reduced Artemia survivorship and hyphal growth of Fusarium during the immature and mature stages. Only mature fruit of two vertebrate-dispersed species reduced hyphal growth of Phoma. Predispersal seed survival increased with toxicity of immature fruit to Artemia during germination and decreased with toxicity to fungi during fruit development. This study suggests that fruit toxicity against generalist natural enemies may be common in Central Panama. These results support the hypothesis that secondary metabolites in fruit have adaptive value and are important in the evolution of fruit-frugivore interactions.

Health Benefits of Vitamins and Secondary Metabolites of Fruits and Vegetables and Prospects To Increase Their Concentrations by Agronomic Approaches

Fruits and vegetables (FAVs) are an important part of the human diet and a major source of biologically active substances such as vitamins and secondary metabolites. The consumption of FAVs remains globally insufficient, so it should be encouraged, and it may be useful to propose to consumers FAVs with enhanced concentrations in vitamins and secondary metabolites. There are basically two ways to reach this target: the genetic approach or the environmental approach. This paper provides a comprehensive review of the results that have been obtained so far through purely agronomic approaches and brings them into perspective by comparing them with the achievements of genetic approaches. Although agronomic approaches offer very good perspectives, the existence of variability of responses suggests that the current understanding of the way regulatory and metabolic pathways are controlled needs to be increased. For this purpose, more in-depth study of the interactions existing between factors (light and temperature, for instance, genetic factors × environmental factors), between processes (primary metabolism and ontogeny, for example), and between organs (as there is some evidence that photooxidative stress in leaves affects antioxidant metabolism in fruits) is proposed.

Comparative behavioural responses of silvereyes (Zosterops lateralis) and European blackbirds (Turdus merula) to secondary metabolites in grapes

Secondary metabolites in fruit are compounds that are not directly associated with plant growth; some are directly related to plant reproductive processes, specifically seed protection and dispersion. There is a complex and species-specific interaction between these plant compounds and their avian seed dispersers. To determine whether two important secondary metabolites in wine grapes – tannins and colour pigments – might be important cues to two of the avian species that forage on wine grapes, and how species-specific this interaction might be, comparative field experiments were run with Australasian silvereyes (Zosterops lateralis) and European blackbirds (Turdus merula). Both species were offered a glucose/fructose artificial nectar with increasing concentrations of grape tannins. In a second experiment, they were offered both green and purple artificial grapes where only the colour differed. Both species showed aversion to tannins; silvereyes appeared to have greater tolerance than blackbirds of tannin concentrations above 5%. In summer no preference was shown between green and purple coloured artificial grapes, but in late autumn blackbirds took only purple grapes whereas silvereyes pecked mostly at green. Links between tannin for seed protection and colour signals to birds are discussed. Colour may cue the two species to different species-specific aspects of fruit nutritive value.

THE EFFECT OF ENVIRONMENTAL FACTORS ON BIOSYNTHESIS OF CAROTENOIDS AND POLYPHENOLICS IN FRUITS AND VEGETABLES: A REVIEW AND PROSPECTS

There is a lack of basic knowledge about the effects of environmental factors on the concentration of secondary metabolites in fruits and vegetables. While metabolic pathways are increasingly better described, the effects of environmental factors remain insufficiently understood and quantified. The objective of this presentation is to review the major hypotheses about the effect of environmental factors on biosynthesis and accumulation of carotenoids and polyphenolic compounds in fruits and vegetables, with the exception of quality of light. Firstly, the primary and the secondary metabolisms are interlinked. Biosynthesis of carotenoids and polyphenolic compounds depends on the carbon status. Secondly, it is now well-established that secondary metabolites play a key-role in adaptation of plants to environmental constraints. More specifically, observations strongly suggest that biotic and abiotic constraints may influence the synthesis of carotenoids through the mediation of Reactive Oxygen Species (ROS). All these hypotheses and the prospects of modelling are presented and discussed from a controlling point of view with our objective being to design innovative cropping techniques for farmers in developed and developing countries which will help them to produce fruits and vegetables with enhanced concentrations in phytochemicals without resorting to GMO. The idea is discussed that organic farming results in fruits and vegetables with increased concentrations in polyphenolics, and possibly carotenoids, contributing to health benefits, because they are more stressed. (Resume d'auteur)

GENES ENCODING FLAVONOID 3'-HYDROXYLASE IN APPLE AND THEIR TAGGED MOLECULAR MARKERS

Flavonoids are important secondary metabolites in fruits, and contribute to both fruit quality and human health. Characterization of genes involved in flavonoid synthesis is critical for genetic modification of the amount and composition of flavonoids in fruits. Here, we report on the isolation of genes encoding flavonoid 3'-hydroxylase (F3'H) in apple. Three genes, designated MdF3'H1, MdF3'H2-1, and MdF3'H2-2 were identified from cv. 'GoldRush' based on EST database assembly and BAC library screening. Each MdF3'H gene is composed of 3 exons and 2 introns and contains an open reading frame of 1,536 bp encoding a putative protein of 511 amino acids. MdF3'H1 shows 90% nucleotide sequence identities in coding region sequence with either MdF3'H2-1 or MdF3'H2-2. MdF3'H2-1 and MdF3'H2-2 share an identity of 99% nucleotide sequence identity in the coding region, and were identified to be alleles. An indel ∼540 bp in size was identified in the first intron of MdF3'H2-1 and MdF3'H2-2, and successfully used to develop gene-tag STS for MdF3'H2 gene. In addition, a gene-tag STS was also developed for MdF3'H1 based on a (CT)n repeat in the second intron.

Quantitative metabolic profiling by 1-dimensional 1H-NMR analyses: application to plant genetics and functional genomics.

Metabolic profiling by 1-dimensional (1-D) 1H-nuclear magnetic resonance (NMR) was tested for absolute quantification of soluble sugars, organic acids, amino acids and some secondary metabolites in fruit, roots and leaves. The metabolite responsible for each peak of the 1H-NMR spectra was identified from spectra of pure compounds. Peak identity was confirmed by the addition of a small amount of commercially-available pure substance. 1H-NMR spectra acquisition was automated. 1H-NMR absolute quantification was performed with a synthesised electronic reference signal and validated by comparison with enzymatic or HPLC analyses; the correlation coefficients between 1H-NMR data and enzymatic or HPLC data were highly significant. Depending on the species and tissues, 14-17 metabolites could be quantified with 15-25 min acquisition time. The detection limit was approximately 1-9 µg in the NMR tube, depending on the compound. Quantitative data were used for (1) a genetic study of strawberry fruit quality, (2) a functional study of tomato transformants overexpressing hexokinase and (3) a study of Arabidopsis phosphoenolpyruvate carboxylase transformants with several lines showing decreased activity of the enzyme. Biochemical phenotyping of the fruits of a strawberry offspring allowed the detection of quantitative trait loci (QTL) controlling fruit quality. Comparison of the roots of wild types and hexokinase tomato transformants using principal component analysis of metabolic profiles revealed that environmental factors, i.e. culture conditions, can significantly modify the metabolic status of plants and thus hide or emphasise the expression of a given genetic background. The decrease in phosphoenolpyruvate carboxylase activity (up to 75%) in Arabidopsis transformants impacted on the metabolic profiles without compromising plant growth, thus supporting the idea that the enzyme has a low influence on the carbon flux through the anaplerotic pathway.

Fruits, frugivores and the evolutionary arms race.

Fruits, frugivores and the evolutionary arms race.

Within population variation and interrelationships between morphology, nutritional content, and secondary compounds of Rhamnus alaternus fruits.

• We studied within-species variation in and interrelations among morphological and chemical traits of ripe Mediterranean buckthorn ( Rhamnus alaternus ) fruit, a bird-dispersed species. • Principal component analysis revealed that larger fruits tended to be relatively rich in nonstructural carbohydrates (NSC), water and P but poor in protein and most minerals. Small fruits tended to be relatively rich in protein, structural carbohydrates, K and Zn while intermediate size fruits tended to be rich in lipids, Mg and Ca. Variation in chemical traits (organic compounds and minerals) was typically much higher than in morphological traits (e.g. fruit size) with the exception of NSC and water content, which varied little. This discrepancy might be explained by differences in environmental conditions between plant microsites that imposed greater variability on fruit nutrient composition than on fruit-morphological traits; and by lower selective pressure by birds on fruit chemical traits than on morphological traits. • Secondary metabolite (emodin) concentration was positively correlated with concentrations of NSC, supporting the nutrient/toxin titration model, which predicts that high levels of secondary metabolites in fruits should be off set by high nutritional rewards for dispersers. • Emodin concentration in leaves was much higher than in fruit pulp, which may indicate its differential adaptive roles in seed dispersal and against herbivores.