Metabolic Variation among Fruits of Different Chili Cultivars (Capsicum spp.) Using HPLC/MS

Chilies are widely cultivated for their rich metabolic content, especially capsaicinoids. In our study, we determined individual sugars, organic acids, capsaicinoids, and total phenolic content in pericarp, placenta, and seeds of Capsicum annuum L., Capsicum chinense Jacq. and Capsicum baccatum L. by HPLC/MS. Dry weight varied in the cultivar ‘Cayenne’, with the first fruit having the lowest dry weight, with 4.14 g. The total sugar content and organic acid content did not vary among the fruits of all three cultivars. The cultivar ‘Cayenne’ showed differences in total phenolic and capsaicinoid content between fruits in the placenta, with the first fruit having the highest content of total phenolics (27.85 g GAE/kg DW) and total capsaicinoids (16.15 g/kg DW). Of the three cultivars studied, the cultivar ‘Habanero Orange’ showed the least variability among fruits in terms of metabolites. The content of dihydrocapsaicin, nordihydrocapsaicin, homocapsaicin, and homodihydrocapsaicin in the seeds of the second fruit was higher than that of the first fruit of the cultivar ‘Bishop Crown’. The results of our study provided significant insight into the metabolomics of individual fruits of the same chili plant. We have thus increased our understanding of how certain metabolites are distributed between fruits at different levels of the same plant and different parts of the fruit. This could be further investigated when chilies are exposed to different environmental stresses.

Methodology for Single Bee and Bee Brain 1H-NMR Metabolomics

The feasibility of metabolomic 1H NMR spectroscopy is demonstrated for its potential to help unravel the complex factors that are impacting honeybee health and behavior. Targeted and non-targeted 1H NMR metabolic profiles of liquid and tissue samples of organisms could provide information on the pathology of infections and on environmentally induced stresses. This work reports on establishing extraction methods for NMR metabolic characterization of Apis mellifera, the European honeybee, describes the currently assignable aqueous metabolome, and gives examples of diverse samples (brain, head, body, whole bee) and biologically meaningful metabolic variation (drone, forager, day old, deformed wing virus). Both high-field (600 MHz) and low-field (80 MHz) methods are applicable, and 1H NMR can observe a useful subset of the metabolome of single bees using accessible NMR instrumentation (600 MHz, inverse room temperature probe) in order to avoid pooling several bees. Metabolite levels and changes can be measured by NMR in the bee brain, where dysregulation of metabolic processes has been implicated in colony collapse. For a targeted study, the ability to recover 10-hydroxy-2-decenoic acid in mandibular glands is shown, as well as markers of interest in the bee brain such as GABA (4-aminobutyrate), proline, and arginine. The findings here support the growing use of 1H NMR more broadly in bees, native pollinators, and insects.

Effect of Environmental Factors on Plectranthus Neochilus Volatile Composition: A GC-MS-Based Metabolomics Approach

Plectranthus neochilus Schltr. is an aromatic species, commonly used for digestive, antispasmodic, and analgesic purposes. Although many studies have reported the chemical composition of its essential oil, variations in the volatile profile were observed, which may be due to multiple factors linked to growth and field conditions. In order to detect metabolic variations in this species, we employed a GC-MS-based untargeted metabolomics approach analyzing samples of four P. neochilus individuals collected over a year. From all analyses, 24 mass features were detected and 21 were identified according to their respective chromatographic peaks. All features varied among samples, particularly (2E)-hexenal, 3-octanone and δ-3-carene, which showed the highest coefficient of variation percentage in our study. Although the four individuals presented the same peaks in the chromatograms, significant differences in the intensity of specific mass features were detected between individuals throughout the year. Time of sampling did not affect P. neochilus volatile composition; the chemical profile remained constant throughout the day. Seasonal trends were observed for the species. Winter months coincided with a drop in the intensity of most components. Air temperature showed a positive correlation with some feature intensities, while myrcene and α-thujene resulted in a positive and a negative correlation with rainfall, respectively. This study was the first attempt to correlate metabolic variation and environmental factors in P. neochilus. Our approach was successful in identifying the composition and variation of the headspace volatiles of P. neochilus leaves.

Tissue-Dependent Variation Profiles of Tea Quality-Related Metabolites in New Shoots of Tea Accessions

Several metabolites define tea quality in new tea shoots composed of leaf and stem. To improve tea quality for breeding, it is important to understand the tissue-dependent genetic mechanisms and metabolic network responsible for the profile of tea quality-related metabolites. We analyzed the volatiles and specialized metabolites as the tea quality-related metabolites in leaves and stems of new shoots in 30 tea accessions to understand the tissue variation and network between tea quality-related metabolites. Our results provided the tissue-dependent variation network in the tea quality-related metabolites, including volatiles in new leaves and stems in tea accessions. Each volatile content in tea accessions showed the coefficient of variation ranging from 58.7 to 221.9% and 54.2 to 318.3% in new leaves and new stems, respectively. The accumulation pattern of tea quality-related metabolites in new leaves and stems varied depending on the accession. When comparing tea genetic populations, the profile of tea quality-related metabolites of new leaves, but not new stems, was the key to distinguishing tea genetic populations by chemical indicators. We described the network between tea quality-related metabolites, especially the dense network in new leaves. These results also will provide the key information for metabolic engineering and the selection of breeding materials in tea plants based on the tea quality-related metabolites and aid in understanding their molecular mechanisms and network of metabolic variation.

Latex Metabolome of Euphorbia Species: Geographical and Inter-Species Variation and its Proposed Role in Plant Defense against Herbivores and Pathogens

Based on the hypothesis that the variation of the metabolomes of latex is a response to selective pressure and should thus be affected differently from other organs, their variation could provide an insight into the defensive chemical selection of plants. Metabolic profiling was used to compare tissues of three Euphorbia species collected in diverse regions. The metabolic variation of latexes was much more limited than that of other organs. In all the species, the levels of polyisoprenes and terpenes were found to be much higher in latexes than in leaves and roots of the corresponding plants. Polyisoprenes were observed to physically delay the contact of pathogens with plant tissues and their growth. A secondary barrier composed of terpenes in latex and in particular, 24-methylenecycloartanol, exhibited antifungal activity. These results added to the well-known role of enzymes also present in latexes, show that these are part of a cooperative defense system comprising biochemical and physical elements.

Effect of the Andean Geography and Climate on the Specialized Metabolism of Its Vegetation: The Subtribe Espeletiinae (Asteraceae) as a Case Example

The Andean mountains are ‘center stage’ to some of the most spectacular examples of plant diversifications, where geographic isolation and past climatic fluctuations have played a major role. However, the influence of Andean geography and climate as drivers of metabolic variation in Andean plants is poorly elucidated. Here, we studied the influence of those factors on the metabolome of the subtribe Espeletiinae (Asteraceae) using liquid chromatography coupled to high-resolution mass spectrometry data of over two hundred samples from multiple locations. Our results demonstrate that metabolic profiles can discriminate Espeletiinae taxa at different geographic scales, revealing inter- and intraspecific metabolic variations: at the country level, segregation between Colombian and Venezuelan taxa was observed; regionally, between páramo massifs; and locally, between páramo complexes. Metabolic differences in Espeletiinae were mainly explained by geographic isolation, although differences in taxonomic genera, temperature, and elevation, were also important. Furthermore, we found that different species inhabiting the same páramo complex showed stronger chemical similarities than the same species at different locations, corroborating that geographic isolation represents the main driver of metabolic change in Espeletiinae. The current study serves as a starting point to fill in the gaps in how Andean geography and climate have shaped the metabolism of its vegetation and reveal the potential of untargeted metabolomics to study the environmental physiology of plants.

The Assessment and the Within-Plant Variation of the Morpho-Physiological Traits and VOCs Profile in Endemic and Rare Salvia ceratophylloides Ard. (Lamiaceae)

Salvia ceratophylloides (Ard.) is an endemic and rare plant species recently rediscovered as very few individuals at two different Southern Italy sites. The study of within-plant variation is fundamental to understand the plant adaptation to the local conditions, especially in rare species, and consequently to preserve plant biodiversity. Here, we reported the variation of the morpho-ecophysiological and metabolic traits between the sessile and petiolate leaf of S. ceratophylloides plants at two different sites for understanding the adaptation strategies for surviving in these habitats. The S. ceratophylloides individuals exhibited different net photosynthetic rate, maximum quantum yield, light intensity for the saturation of the photosynthetic machinery, stomatal conductance, transpiration rate, leaf area, fractal dimension, and some volatile organic compounds (VOCs) between the different leaf types. This within-plant morpho-physiological and metabolic variation was dependent on the site. These results provide empirical evidence of sharply within-plant variation of the morpho-physiological traits and VOCs profiles in S. ceratophylloides, explaining the adaptation to the local conditions.

The Assessment and the Within-Plant Variation of the Morpho-Physiological Traits and VOCs Profile in Endemic and Rare Salvia ceratophylloides Ard. (Lamiaceae)

Salvia ceratophylloides (Ard.) is an endemic, rare, threatened plant species recently rediscovered in very few individuals in two different sites of South Italy. The study of within-plant variation more than among-plant one is fundamental to understand the plant adaptation to the local conditions, especially in rare species, and consequently to preserve plant biodiversity. Here, we reported the variation of the morpho-ecophysiological and metabolic traits between the sessile and petiolate leaf of S. ceratophylloides plants in two different sites for understanding the adaptation strategies for surviving in these habitats. The S. ceratophylloides individuals exhibited different net photosynthetic rate, maximum quantum yield, light intensity for the saturation of the photosynthetic machinery, stomatal conductance, transpiration rate, leaf area, fractal dimension and some VOCs between the different leaf types. This within-plant morpho-physiological and metabolic variation was depended on the site. These results provide empirical evidence of sharply within-plant variation of the morpho-physiological traits and VOCs profiles in S. ceratophylloides which could be because of adaptation to the local conditions.