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A. Gargallo-Garriga, J. Sardans, J. Llusià, G. Peguero, D. Asensio, R. Ogaya, I. Urbina, L. Van Langenhove, L. T. Verryckt, E. A. Courtois, C. Stahl, O. Grau, O. Urban, I. A. Janssens , P. Nolis, M. Pérez-Trujillo, T. Parella and J. Peñuelas
Molecules, 25, 3960 (2020). DOI
31P-NMR Metabolomics Revealed Species-Specific Use of Phosphorous in Trees of a French Guiana Rainforest
Productivity of tropical lowland moist forests is often limited by availability and functional allocation of phosphorus (P) that drives competition among tree species and becomes a key factor in determining forestall community diversity. We used non-target 31P-NMR metabolic profiling to study the foliar P-metabolism of trees of a French Guiana rainforest. The objective was to test the hypotheses that P-use is species-specific, and that species diversity relates to species P-use and concentrations of P-containing compounds, including inorganic phosphates, orthophosphate monoesters and diesters, phosphonates and organic polyphosphates. We found that tree species explained the 59% of variance in 31P-NMR metabolite profiling of leaves. A principal component analysis showed that tree species were separated along PC 1 and PC 2 of detected P-containing compounds, which represented a continuum going from high concentrations of metabolites related to non-active P and P-storage, low total P concentrations and high N:P ratios, to high concentrations of P-containing metabolites related to energy and anabolic metabolism, high total P concentrations and low N:P ratios. These results highlight the species-specific use of P and the existence of species-specific P-use niches that are driven by the distinct species-specific position in a continuum in the P-allocation from P-storage compounds to P-containing molecules related to energy and anabolic metabolism.
M. Palomino-Schätzlein, Y. Wang, A. D. Brailsford, T. Parella, D. A. Cowan, C. Legido-Quigley, and M. Pérez-Trujillo
Analytical Chemistry, 89, 8343 (2017). DOI
Direct Monitoring of Exogenous γ‑Hydroxybutyric Acid in Body Fluids by NMR Spectroscopy
γ-Hydroxybutyric acid (GHB) is a popular drug increasingly associated with cases of drug-facilitated sexual assault (DFSA). Currently, expanding procedures of analysis and having forensic evidence of GHB intake in a long term are mandatory. Up to now, most studies have been performed using GC/MS and LC-MS as analytical platforms, which involve significant manipulation of the sample and, often, indirect measurements. In this work, procedures used in NMR-based metabolomics were applied to a GHB clinical trial on urine and serum. Detection, identification, and quantification of the drug by NMR methods were surveyed, as well as the use of NMR-based metabolomics for the search of potential surrogate biomarkers of GHB consumption. Results demonstrated the suitability of NMR spectroscopy, as a robust nondestructive technique, to fast and directly monitor (detect, identify, and quantify) exogenous GHB in almost intact body fluids and its high potential in the search for metabolites associated with GHB intake.
A. Gargallo-Garriga , S. J. Wright, J. Sardans, M. Pérez-Trujillo, M. Oravec, K. Večeřová, O. Urban, M. Fernández-Martínez, T. Parella and J. Peñuelas
PLoS One, 16, 78 (2017). DOI
Long-term fertilization determines different metabolomic profiles and responses in saplings of three rainforest tree species with different adult canopy position
We studied the foliar metabolome of saplings of three abundant tree species in a 15 year field NPK fertilization experiment in a Panamanian rainforest. The largest differences were among species and explained 75% of overall metabolome variation. The saplings of the large canopy species, Tetragastris panamensis , had the lowest concentrations of all identi- fied amino acids and the highest concentratio ns of most identified secondary compounds. The saplings of the “ mid canopy” species, Alseis blackiana , had the highest concentratio ns of amino acids coming from the biosynthesis pathways of glycerate-3P, oxaloacetate and α - ketoglutarate, and the saplings of the low canopy species, Heisteria concinna , had the high- est concentratio ns of amino acids coming from the pyruvate synthesis pathways.
A. Gargallo-Garriga, J. Sardans, M. Pérez-Trujillo, A. Guenther, J. Llusià, L. Rico, J. Terradas, G. Farré-Armengol, I. Filella, T. Parella and J. Peñuelas
BMC Plant Biology, 16, 78-89 (2016). DOI
Shifts in plant foliar and floral metabolomes in response to the suppression of the associated microbiota
The phyllospheric microbiota is assumed to play a key role in the metabolism of host plants. Its role in determining the epiphytic and internal plant metabolome, however, remains to be investigated. We analyzed profiles of the epiphytic and internal metabolomes of the leaves and flowers of Sambucus nigra with and without external antibiotic treatment application. These results suggest that anaerobic and/or facultative anaerobic bacteria were present in high numbers in the phyllosphere and in the apoplasts of S. nigra. The results also show that microbial communities play a significant role in the metabolomes of plant organs and could have more complex and frequent mutualistic, saprophytic, and/or parasitic relationships with internal plant metabolism than currently assumed.
A. Gargallo-Garriga, J. Sardans, M. Pérez-Trujillo, M. Oravec, O. Urban, A. Jentsch, J. Kreyling, C. Beierkuhnlein, T. Parella, and J. Peñuelas
New Phytologist, 207, 591-603 (2015). DOI
Warming differentially influences the effects of drought on stoichiometry in metabolomics in shoots and roots
The stoichiometric and metabolomic responses of plants to warming strongly depend on water availability, and the response differs in shoots and roots. Warming under drought conditions stimulates root primary metabolic activity more strongly than under drought alone. Compared with drought alone, shoots under simultaneous warming and drought shifted their osmoprotective and antistress strategies by the down- and upregulation of the synthesis of various secondary metabolites and by the activation of some primary metabolic pathways. Our results thus demonstrated different metabolomic expressions in different parts of the plant and a large plasticity in the responses to environmental changes.
A. Gargallo-Garriga, J. Sardans, M. Pérez-Trujillo, A. Rivas-Ubach, M. Oravec, K. Vecerova, O. Urban, A. Jentsch, J. Kreyling, C. Beierkuhnlein, T. Parella and J. Peñuelas
Scientific Reports, 4, 6829 (2014). DOI
Opposite metabolic responses of shoots and roots to drought
Shoots and roots have different metabolomes and nutrient and elemental stoichiometries, the shoot metabolome is much more variable among species and seasons than is the root metabolome and the metabolic response of shoots to drought contrasts with that of roots; shoots decrease their growth metabolism (lower concentrations of sugars, amino acids, nucleosides, N, P, and K), and roots increase it in a mirrored response.
A. Rivas-Ubach, A. Gargallo-Garriga, J. Sardans, M. Oravec, L. Mateu-Castell, M. Pérez-Trujillo, T. Parella, R. Ogaya, O. Urban and J. Peñuelas
New Phytologist, 202, 874 (2014). DOI
Drought enhances folivory by shifting foliar metabolomes in Quercus ilex trees
The present study is an example of how coupling stoichiometric with metabolomic techniques is useful for identifying the molecular responses of plants to changing environmental conditions such as drought, for understanding the mechanisms and functions that underlie the responses of plants to these new conditions and for interpreting the implications of drought for trophic webs. Foliar N : P ratios of Q. ilex did not significantly change between seasons and were not lowest in spring, the growing season, as expected from the growth rate hypothesis. Foliar K concentrations, however, did change, demonstrating the importance of K in natural summer droughts. The lack of significant variation in N and P concentrations in Q. ilex may be attributable to the buffering effect of lignotubers in this woody species. Moderate experimental drought increased the concentrations of sugars and polyphenolic compounds with antioxidant function in the leaves of Q. ilex.
A. Rivas-Ubach, M. Perez-Trujillo, J. Sardans, A. Gargallo-Garriga, T. Parella and J. Peñuelas
Methods in Ecology and Evolution, 4, 464-473 (2013). DOI
Ecometabolomics: Optimized NMR-based procedures
Here we propose a NMR‐based protocol for ecometabolomic studies that provides an unbiased overview of the metabolome of an organism, including polar and nonpolar metabolites. This protocol is aimed to facilitate the analysis of many samples, as typically required in ecological studies. In addition to NMR fingerprinting, it identifies metabolites for generating metabolic profiles applying strategies of elucidation of small molecules typically used in natural‐product research, and allowing the identification of secondary and unknown metabolites. We also provide a detailed description to obtain the numerical data from the 1H‐NMR spectra needed to perform the statistical analyses.
M. Pérez-Trujillo, J.C. Lindon, T. Parella, J.K. Nicholson, H.C. Keun, and T.J. Athersuch
Anal. Chem., 84, 2868-2874 (2012). DOI
Chiral Metabonomics: 1H NMR-based enantiospecific differentiation of metabolites in human urine via co-solvation with beta-cyclodextrin
Differences in molecular chirality remain an important issue in drug metabolism and pharmacokinetics for the pharmaceutical industry and regulatory authorities, and chirality is an important feature of many endogenous metabolites. We present a method for the rapid, direct differentiation and identification of chiral drug enantiomers in human urine without pretreatment of any kind. Using the well-known anti-inflammatory chemical ibuprofen as one example, we demonstrate that the enantiomers of ibuprofen and the diastereoisomers of one of its main metabolites, the glucuronidated carboxylate derivative, can be resolved by 1H NMR spectroscopy as a consequence of direct addition of the chiral cosolvating agent (CSA) β-cyclodextrin (βCD). This approach is simple, rapid, and robust, involves minimal sample manipulation, and does not require derivatization or purification of the sample. In addition, the method should allow the enantiodifferentiation of endogenous chiral metabolites, and this has potential value for differentiating metabolites from mammalian and microbial sources in biofluids. From these initial findings, we propose that more extensive and detailed enantiospecific metabolic profiling could be possible using CSA-NMR spectroscopy than has been previously reported.
J. Sardans, A. Gargallo-Garriga, M. Pérez-Trujillo, T. Parella, R. Seco, I. Filella and J. Peñuelas
Plant Biology, 16, 395 (2014). DOI
Metabolic responses of Quercus ilex seedlings to wounding analysed with nuclear magnetic resonance profiling
The study shows the rapid metabolic response of Q. ilex in defending its leaves, based on a rapid increase in the production of quinic acid, quercitol and choline. The results also confirm the suitability of 1H NMR-based metabolomic profiling studies to detect global metabolome shifts after wounding stress in tree leaves, and therefore its suitability in ecometabolomic studies.
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