Exploring the Interplay between Metabolic Pathways and Taxane Production in Elicited Taxus baccata Cell Suspensions

Taxus cell cultures are a reliable biotechnological source of the anticancer drug paclitaxel. However, the interplay between taxane production and other metabolic pathways during elicitation remains poorly understood. In this study, we combined untargeted metabolomics and elicited Taxus baccata cell cultures to investigate variations in taxane-associated metabolism under the influence of 1 µM coronatine (COR) and 150 µM salicylic acid (SA). Our results demonstrated pleiotropic effects induced by both COR and SA elicitors, leading to differential changes in cell growth, taxane content, and secondary metabolism. Metabolite annotation revealed significant effects on N-containing compounds, phenylpropanoids, and terpenoids. Multivariate analysis showed that the metabolomic profiles of control and COR-treated samples are closer to each other than to SA-elicited samples at different time points (8, 16, and 24 days). The highest level of paclitaxel content was detected on day 8 under SA elicitation, exhibiting a negative correlation with the biomarkers kauralexin A2 and taxusin. Our study provides valuable insights into the intricate metabolic changes associated with paclitaxel production, aiding its potential optimization through untargeted metabolomics and an evaluation of COR/SA elicitor effects

Dataset on the effects of different pre-harvest factors on the metabolomics profile of lettuce (Lactuca sativa l.) leaves

The study of the relationship between cultivated plants and environmental factors can provide information ranging from a deeper understanding of the plant biological system to the development of more effective management strategies for improving yield, quality, and sustainability of the produce. In this article, we present a comprehensive metabolomics dataset of two phytochemically divergent lettuce (Lactuca sativa L.) butterhead varieties under different growing conditions. Plants were cultivated in hydroponics in a growth chamber with ambient control. The pre-harvest factors that were independently investigated were light intensity (two levels), the ionic strength of the nutrient solutions (three levels), and the molar ratio of three macroelements (K, Mg, and Ca) in the nutrient solution (three levels). We used an untargeted, mass-spectrometry-based approach to characterize the metabolomics profiles of leaves harvested 19 days after transplant. The data revealed the ample impact on both primary and secondary metabolism and its range of variation. Moreover, our dataset is useful for uncovering the complex effects of the genotype, the environmental factor(s), and their interaction, which may deserve further investigation

Intraspecific variability largely affects the leaf metabolomics response to isosmotic macrocation variations in two divergent lettuce (Lactuca sativa L.) varieties

Mineral elements are essential for plant growth and development and strongly affect crop yield and quality. To cope with an everchanging environment, plants have developed specific responses to combined nutrient variations. In this work, we investigated the effects of multifactorial treatments with three macrocations (K, Ca, and Mg) on lettuce (Lactuca sativa L.) varieties that strongly diverge in leaf pigmentation (full red or green). Specifically, we monitored main leaf parameters and metabolomics profiles of hydroponically grown plants fed with isosmotic nutrient solutions that have different proportions of macroelements. The result revealed a high biochemical plasticity of lettuce, significantly affected by the genotype, the nutrient solution, and their interaction. Our work also provided evidence and insights into the different intraspecific responses to multifactorial variation of macrocations, with two varieties having distinct strategies to metabolically respond to nutrient variation. Overall, plant adaptive mechanisms increased the phytochemical diversity between the varieties both among and within the main classes of plant secondary metabolites. Finally, our work also implies that the interaction of a pre-existing phytochemical diversity with the management of multiple mineral elements can offer added health-related benefits to the edible product specific to the variety

Beyond the Visible and Below the Peel: How UV-B Radiation Influences the Phenolic Profile in the Pulp of Peach Fruit. A Biochemical and Molecular Study

In the last decades, UV-B radiation has attracted attention due to its potential to increase nutraceutical values of fruit and vegetables, especially by inducing the accumulation of phenolics in a structure-dependent way. However, most current studies have investigated the UV-B-driven changes only in the peel or focusing on individual phenolic classes. Adopting an \u201c-omics\u201d approach, this work aimed to deepen the knowledge about the effects of UV-B radiation on the phenolic profile in the pulp of peach fruit. Based on these considerations, melting flesh yellow peaches (Prunus persica L., cv. Fairtime) were subjected to either a 10- or 60-min UV-B treatment (1.39 and 8.33 kJ m\u20132, respectively), and sampled at different time points from the exposure. A UHPLC-ESI/QTOF-MS analysis coupled with a phenolics-specific database for the annotation of compounds and a multivariate discriminant analysis revealed a marked effect of UV-B radiation on the phenolic profiles of peach pulp. Particularly, a general, transient increase was observed after 24 h from the irradiation, especially for flavanols, flavonols, and flavones. Such behavior diverges from what was observed in the peel, where an overall increase of phenolics was observed after 36 h from the irradiation. Concerning the flavonols in the pulp, UV-B exposure stimulated a specific accumulation of isorhamnetin and kaempferol derivatives, with variations imposed by the different sugar moiety bound. Anthocyanins, which were the second most abundant flavonoid group after flavonols, displayed a general decrease after 36 h that was not attributable to specific molecules. The UV-B treatments also increased the glycoside/aglycone ratio of flavonols and anthocyanins after 24 h, by increasing the glycoside concentration of both, flavonols and anthocyanins, and decreasing the aglycone concentration of anthocyanins. In support of the biochemical results, targeted gene expression analysis by RT-qPCR revealed an UV-B-induced activation of many genes involved in the flavonoid pathway, e.g., CHS, F3H, F3\u2032H, DFR, as well as some MYB transcription factors and few genes involved in the UV-B perception. Generally, all the flavonoid-related and MYB genes showed a transient UV-B dose-dependent activation after 6 h from the irradiation, similarly to what was observed in the peel

A Microbial-Based Biostimulant Enhances Sweet Pepper Performance by Metabolic Reprogramming of Phytohormone Profile and Secondary Metabolism

Microbial-based biostimulants can improve crop productivity by modulating cell metabolic pathways including hormonal balance. However, little is known about the microbial-mediated molecular changes causing yield increase. The present study elucidates the metabolomic modulation occurring in pepper (Capsicum annuum L.) leaves at the vegetative and reproductive phenological stages, in response to microbial-based biostimulants. The arbuscular mycorrhizal fungi Rhizoglomus irregularis and Funneliformis mosseae, as well as Trichoderma koningii, were used in this work. The application of endophytic fungi significantly increased total fruit yield by 23.7% compared to that of untreated plants. Multivariate statistics indicated that the biostimulant treatment substantially altered the shape of the metabolic profile of pepper. Compared to the untreated control, the plants treated with microbial biostimulants presented with modified gibberellin, auxin, and cytokinin patterns. The biostimulant treatment also induced secondary metabolism and caused carotenoids, saponins, and phenolic compounds to accumulate in the plants. Differential metabolomic signatures indicated diverse and concerted biochemical responses in the plants following the colonization of their roots by beneficial microorganisms. The above findings demonstrated a clear link between microbial-mediated yield increase and a strong up-regulation of hormonal and secondary metabolic pathways associated with growth stimulation and crop defense to environmental stresses

Foliar application of different vegetal-derived protein hydrolysates distinctively modulates tomato root development and metabolism

Despite the scientific evidence supporting their biostimulant activity, the molecular mechanism(s) underlying the activity of protein hydrolysates (PHs) and the specificity among different products are still poorly explored. This work tested five different protein hydrolysates, produced from different plant sources using the same enzymatic approach, for their ability to promote rooting in tomato cuttings following quick dipping. Provided that all the different PHs increased root length (45\u201393%) and some of them increased root number (37\u201356%), untargeted metabolomics followed by multivariate statistics and pathway analysis were used to unravel the molecular processes at the basis of the biostimulant activity. Distinct metabolomic signatures could be found in roots following the PHs treatments. In general, PHs shaped the phytohormone profile, modulating the complex interaction between cytokinins and auxins, an interplay playing a pivotal role in root development, and triggered a down accumulation of brassinosteroids. Concerning secondary metabolism, PHs induced the accumulation of aliphatic glucosinolates, alkaloids, and phenylpropanoids, potentially eliciting crop resilience to stress conditions. Here, we confirm that PHs may have a hormone-like activity, and that their application can modulate plant growth, likely interfering with signaling processes. Noteworthy, the heterogenicity of the botanical origin supported the distinctive and peculiar metabolomic responses we observed across the products tested. While supporting their biostimulant activity, these findings suggest that a generalized crop response to PHs cannot be defined and that specific effects are rather to be investigated

Foliar and root comparative metabolomics and phenolic profiling of Micro-Tom tomato (Solanum lycopersicum L.) plants associated with a gene expression analysis in response to short daily UV treatments

Tomato (Solanum lycopersicum L.) is globally recognised as a high-value crop both for commercial profit and nutritional benefits. In contrast to the extensive data regarding the changes in the metabolism of tomato fruit exposed to UV radiation, less is known about the foliar and root metabolome. Using an untargeted metabolomic approach through UHPLC-ESI-QTOF-MS analysis, we detected thousands of metabolites in the leaves (3000) and roots (2800) of Micro-Tom tomato plants exposed to 11 days of short daily UV radiation, applied only on the aboveground organs. Multivariate statistical analysis, such as OPLS-DA and volcano, were performed to allow a better understanding of the modifications caused by the treatment. Based on the unexpected modulation to the secondary metabolism, especially the phenylpropanoid pathway, of which compounds were down and up accumulated respectively in leaves and roots of treated plants, a phenolic profiling was carried out for both organs. The phenolic profile was associated with a gene expression analysis to check the transcription trend of genes involved in the UVR8 signalling pathway and the early steps of the phenolic biosynthesis. The retention of the modifications at metabolic and phenolic levels was also investigated 3 days after the UV treatment, showing a prolonged effect on the modulation once the UV treatment had ceased

Nutraceutical Profiles of Two Hydroponically Grown Sweet Basil Cultivars as Affected by the Composition of the Nutrient Solution and the Inoculation With Azospirillum brasilense

Sweet basil (Ocimum basilicum L.) is one of the most produced aromatic herbs in the world, exploiting hydroponic systems. It has been widely assessed that macronutrients, like nitrogen (N) and sulfur (S), can strongly affect the organoleptic qualities of agricultural products, thus influencing their nutraceutical value. In addition, plant-growth-promoting rhizobacteria (PGPR) have been shown to affect plant growth and quality. Azospirillum brasilense is a PGPR able to colonize the root system of different crops, promoting their growth and development and influencing the acquisition of mineral nutrients. On the bases of these observations, we aimed at investigating the impact of both mineral nutrients supply and rhizobacteria inoculation on the nutraceutical value on two different sweet basil varieties, i.e., Genovese and Red Rubin. To these objectives, basil plants have been grown in hydroponics, with nutrient solutions fortified for the concentration of either S or N, supplied as SO42\u2013 or NO3\u2013, respectively. In addition, plants were either non-inoculated or inoculated with A. brasilense. At harvest, basil plants were assessed for the yield and the nutraceutical properties of the edible parts. The cultivation of basil plants in the fortified nutrient solutions showed a general increasing trend in the accumulation of the fresh biomass, albeit the inoculation with A. brasilense did not further promote the growth. The metabolomic analyses disclosed a strong effect of treatments on the differential accumulation of metabolites in basil leaves, producing the modulation of more than 400 compounds belonging to the secondary metabolism, as phenylpropanoids, isoprenoids, alkaloids, several flavonoids, and terpenoids. The primary metabolism that resulted was also influenced by the treatments showing changes in the fatty acid, carbohydrates, and amino acids metabolism. The amino acid analysis revealed that the treatments induced an increase in arginine (Arg) content in the leaves, which has been shown to have beneficial effects on human health. In conclusion, between the two cultivars studied, Red Rubin displayed the most positive effect in terms of nutritional value, which was further enhanced following A. brasilense inoculation

A combined phenotypic and metabolomic approach for elucidating the biostimulant action of a plant-derived protein hydrolysate on tomato grown under limited water availability

Plant-derived protein hydrolysates (PHs) are an important category of biostimulants able to increase plant growth and crop yield especially under environmental stress conditions. PHs can be applied as foliar spray or soil drench. Foliar spray is generally applied to achieve a relatively short-term response, whereas soil drench is used when a long-term effect is desired. The aim of the study was to elucidate the biostimulant action of PH application method (foliar spray or substrate drench) on morpho-physiological traits and metabolic profile of tomato grown under limited water availability. An untreated control was also included. A high-throughput image-based phenotyping (HTP) approach was used to non-destructively monitor the crop response under limited water availability (40% of container capacity) in a controlled environment. Moreover, metabolic profile of leaves was determined at the end of the trial. Dry biomass of shoots at the end of the trial was significantly correlated with number of green pixels (R2 = 0.90) and projected shoot area, respectively. Both drench and foliar treatments had a positive impact on the digital biomass compared to control while the photosynthetic performance of the plants was slightly influenced by treatments. Overall drench application under limited water availability more positively influenced biomass accumulation and metabolic profile than foliar application. Significantly higher transpiration use efficiency was observed with PH-drench applications indicating better stomatal conductance. The mass-spectrometry based metabolomic analysis allowed the identification of distinct biochemical signatures in PH-treated plants. Metabolomic changes involved a wide and organized range of biochemical processes that included, among others, phytohormones (notably a decrease in cytokinins and an accumulation of salicylates) and lipids (including membrane lipids, sterols, and terpenes). From a general perspective, treated tomato plants exhibited an improved tolerance to reactive oxygen species (ROS)-mediated oxidative imbalance. Such capability to cope with oxidative stress might have resulted from a coordinated action of signaling compounds (salicylic acid and hydroxycinnamic amides), radical scavengers such as carotenoids and prenyl quinones, as well as a reduced biosynthesis of tetrapyrrole coproporphyrins

Metabolic Insights into the Anion-Anion Antagonism in Sweet Basil: Effects of Different Nitrate/Chloride Ratios in the Nutrient Solution

Sweet basil (Ocimum basilicum L.) is a highly versatile and globally popular culinary herb, and a rich source of aromatic and bioactive compounds. Particularly for leafy vegetables, nutrient management allows a more efficient and sustainable improvement of crop yield and quality. In this work, we investigated the effects of balanced modulation of the concentration of two antagonist anions (nitrate and chlorine) in basil. Specifically, we evaluated the changes in yield and leaf metabolic profiles in response to four different NO3-:Cl- ratios in two consecutive harvests, using a full factorial design. Our work indicated that the variation of the nitrate-chloride ratio exerts a large effect on both metabolomic profile and yield in basil, which cannot be fully explained only by an anion-anion antagonist outcome. The metabolomic reprogramming involved different biochemical classes of compounds, with distinctive traits as a function of the different nutrient ratios. Such changes involved not only a response to nutrients availability, but also to redox imbalance and oxidative stress. A network of signaling compounds, including NO and phytohormones, underlined the modeling of metabolomic signatures. Our work highlighted the potential and the magnitude of the effect of nutrient solution management in basil and provided an advancement towards understanding the metabolic response to anion antagonism in plants