Application of GC-MS for the detection of lipophilic compounds in diverse plant tissues

This study reports a GC-QqQ-MS method for the quantification of forty-eight primary metabolites from four major classes (sugars, sugar acids, sugar phosphates, and organic acids) which can be applied to a number of biological systems. The method was validated in terms of linearity, reproducibility and recovery, using both calibration standards and real samples. Additionally, twenty-eight biogenic amines and amino acids were quantified using an established LC-QqQ-MS method. Both GC-QqQ-MS and LC-QqQ-MS quantitative methods were applied to plant extracts from flower and pod tissue of two chickpea (Cicer arietinum L.) cultivars differing in their ability to tolerate salinity, which were grown under control and salt-treated conditions. Statistical analysis was applied to the data sets using the absolute concentrations of metabolites to investigate the differences in metabolite profiles between the different cultivars, plant tissues, and treatments. The method is a significant improvement of present methodology for quantitative GC-MS metabolite profiling of organic acids and sugars, and provides new insights of chickpea metabolic responses to salinity stress. It is applicable to the analysis of dynamic changes in endogenous concentrations of polar primary metabolites to study metabolic responses to environmental stresses in complex biological tissues.

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“…Only arabitol (127%) and quinate (126%) showed slightly higher recoveries as previously reported [15][16][17][18]. For most metabolites recovery …”

Section: Recoverysupporting

confidence: 58%

“…However, for 2-keto gluconic acid and sucrose the overall recoveries were 34% and 49%, respectively. Several sugars, including erlose (135%), raffinose (147%), and melezitose (133%), show a slightly higher recoveries as previously reported [15][16][17][18].…”

Section: Recoverysupporting

confidence: 46%

Quantitative profiling of polar primary metabolites of two chickpea cultivars with contrasting responses to salinity

Dias

Hill

Jayasinghe

et al. 2015

Journal of Chromatography B

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“…Small amounts of authentic standards, at similar amounts to those present in the biological material, should be added to the biological material before extraction, and recovery through the extraction and analytic procedure should be documented. This is rarely done, and those studies that have been performed with plants (Roessner-Tunali et al, 2003;Schauer et al, 2006;Cruz et al, 2008;Arrivault et al, 2009;Lytovchenko et al, 2009) show that losses of metabolites can occur and that these may be avoided by modifying the extraction protocol or analytic approach.…”

Section: Recovery Experimentsmentioning

confidence: 99%

Metabolic Networks: How to Identify Key Components in the Regulation of Metabolism and Growth

2009

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Plants display enormous diversity in their metabolism. Although the biosynthesis and function of the myriad plant metabolites have been studied for decades, we have little understanding of the interactions between metabolites, metabolite signaling, interactions with development, and the role of metabolism in genotype-to-phenotype relationships. Technologies for the analysis of metabolites have made tremendous progress in recent years, both in terms of the number of metabolites that are identified and of throughput. Recent developments allow the construction of metabolic networks and study of the role of these networks in plant growth and development. In this review, we discuss what types of information can be obtained from measurements of metabolites and what requirements they have with respect to the comprehensiveness of coverage and the precision of identification and quantification, and we outline procedures that can be implemented to validate the measurements. We then discuss what sorts of perturbations can be used to disturb metabolic networks, including environmental and physiological treatments, chemicals, reverse genetics, and the use of natural genetic diversity.Plants are the most consummate and sophisticated chemical system in the world. They use light energy to convert CO 2 into carbohydrates in their leaves. They absorb nutrients like nitrate, phosphate, and sulfate via their roots and convert them to amino acids and nucleotides, using light energy in the leaves in the day and energy derived from respiration in leaves in the dark and in nonphotosynthetic tissues. Carbohydrates, amino acids, and nucleotides are then transported to growing tissues, where they are converted into macromolecular cellular components like proteins, nucleic acids, cell walls, pigments, and lipids. Plants also synthesize tens of thousands of secondary metabolites, including phenylpropanoids and flavonoids, terpenoids, glucosinolates, and alkaloids. These have important roles in cellular function, in signaling, and in adaptation to abiotic and biotic stress. Their unique synthetic ability is the result of a highly complex and sophisticated metabolic apparatus. Its complexity and flexibility were already appreciated by the 1980s, as biochemical studies of cellular compartmentation revealed that many basic pathways like glycolysis, the oxidative pentose phosphate pathway, and organic acid metabolism were present in more than one compartment (Lunn, 2007), and the diversity of plant secondary metabolites was unveiled. It was further underlined as genome sequencing uncovered families of genes for enzymes in central metabolism, including starch and Suc synthesis and degradation, glycolysis, many reactions of the tricarboxylic acid cycle and nucleotide degradation, and huge families of genes that encode enzymes that introduce modifications into basic metabolite structures, like the alcohol dehydrogenases, 2-oxoglutarate dioxygenases, acyl transferases, UDP glucosyltransferases, O-methyl transferases, nitrilases, cytochrome P50s, myr...

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“…For the GC-MS analysis, seeds from immature green fruit (approximately 25 DPA) were manually isolated, frozen, and ground in liquid nitrogen, and a 250-mg aliquot was used for extraction. Lipophilic compounds from the same extraction round were determined by GC-MS following the protocol by Lytovchenko et al (2009). Calvin cycle intermediates were determined as described by Arrivault et al (2009).…”

Section: Determination Of Metabolite Levelsmentioning

confidence: 99%

Tomato Fruit Photosynthesis Is Seemingly Unimportant in Primary Metabolism and Ripening But Plays a Considerable Role in Seed Development

et al. 2011

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Fruit of tomato (Solanum lycopersicum), like those from many species, have been characterized to undergo a shift from partially photosynthetic to truly heterotrophic metabolism. While there is plentiful evidence for functional photosynthesis in young tomato fruit, the rates of carbon assimilation rarely exceed those of carbon dioxide release, raising the question of its role in this tissue. Here, we describe the generation and characterization of lines exhibiting a fruit-specific reduction in the expression of glutamate 1-semialdehyde aminotransferase (GSA). Despite the fact that these plants contained less GSA protein and lowered chlorophyll levels and photosynthetic activity, they were characterized by few other differences. Indeed, they displayed almost no differences in fruit size, weight, or ripening capacity and furthermore displayed few alterations in other primary or intermediary metabolites. Although GSA antisense lines were characterized by significant alterations in the expression of genes associated with photosynthesis, as well as with cell wall and amino acid metabolism, these changes were not manifested at the phenotypic level. One striking feature of the antisense plants was their seed phenotype: the transformants displayed a reduced seed set and altered morphology and metabolism at early stages of fruit development, although these differences did not affect the final seed number or fecundity. Taken together, these results suggest that fruit photosynthesis is, at least under ambient conditions, not necessary for fruit energy metabolism or development but is essential for properly timed seed development and therefore may confer an advantage under conditions of stress.

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Application of GC-MS for the detection of lipophilic compounds in diverse plant tissues

Cited by 60 publications

References 56 publications

Quantitative profiling of polar primary metabolites of two chickpea cultivars with contrasting responses to salinity

Quantitative profiling of polar primary metabolites of two chickpea cultivars with contrasting responses to salinity

Metabolic Networks: How to Identify Key Components in the Regulation of Metabolism and Growth

Tomato Fruit Photosynthesis Is Seemingly Unimportant in Primary Metabolism and Ripening But Plays a Considerable Role in Seed Development

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