Application of a high‐throughput HPLC‐MS/MS assay to <i>Arabidopsis</i> mutant screening; evidence that threonine aldolase plays a role in seed nutritional quality

Jander, Georg; Norris, Susan R.; Joshi, Vijay; Fraga, Michele; Rugg, Arthur; Yu, Sheng; Li, Lily; Last, Robert L.

doi:10.1111/j.1365-313x.2004.02140.x

Cited by 122 publications

(128 citation statements)

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“…The analysis method was modified from a protocol described by Jander et al 69 . Chromatography was performed on an Agilent 1200 HPLC system (Agilent Technologies).…”

Section: Methodsmentioning

confidence: 99%

Metabolic cross-feeding via intercellular nanotubes among bacteria

et al. 2015

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Bacteria frequently exchange metabolites by diffusion through the extracellular environment, yet it remains generally unclear whether bacteria can also use cell-cell connections to directly exchange nutrients. Here we address this question by engineering cross-feeding interactions within and between Acinetobacter baylyi and Escherichia coli, in which two distant bacterial species reciprocally exchange essential amino acids. We establish that in a well-mixed environment E. coli, but likely not A. baylyi, can connect to other bacterial cells via membranederived nanotubes and use these to exchange cytoplasmic constituents. Intercellular connections are induced by auxotrophy-causing mutations and cease to establish when amino acids are externally supplied. Electron and fluorescence microscopy reveal a network of nanotubular structures that connects bacterial cells and enables an intercellular transfer of cytoplasmic materials. Together, our results demonstrate that bacteria can use nanotubes to exchange nutrients among connected cells and thus help to distribute metabolic functions within microbial communities.

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“…The analysis method was modified from a protocol described by Jander et al 69 . Chromatography was performed on an Agilent 1200 HPLC system (Agilent Technologies).…”

Section: Methodsmentioning

confidence: 99%

Metabolic cross-feeding via intercellular nanotubes among bacteria

et al. 2015

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“…Amino acids in the diluted methanol extract were directly analyzed by LC-MS/MS. The method described by Jander et al (2004) was used with some modifications. Briefly, chromatography was performed on an Agilent 1200 HPLC system.…”

Section: Lc-ms/ms Analysis Of Aldoximes and Amino Acidsmentioning

confidence: 99%

“…Multiple reaction monitoring (MRM) was used to monitor a parent ion → product ion reaction for each analyte. MRM was chosen as described by Jander et al (2004), except for Arg (m/z 175 → 70) and Lys (m/z 147 → 84). Both Q1 and Q3 quadrupoles were maintained at unit resolution.…”

Section: Lc-ms/ms Analysis Of Aldoximes and Amino Acidsmentioning

confidence: 99%

Two Herbivore-Induced Cytochrome P450 Enzymes CYP79D6 and CYP79D7 Catalyze the Formation of Volatile Aldoximes Involved in Poplar Defense

et al. 2013

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Aldoximes are known as floral and vegetative plant volatiles but also as biosynthetic intermediates for other plant defense compounds. While the cytochrome P450 monooxygenases (CYP) from the CYP79 family forming aldoximes as biosynthetic intermediates have been intensively studied, little is known about the enzymology of volatile aldoxime formation. We characterized two P450 enzymes, CYP79D6v3 and CYP79D7v2, which are involved in herbivore-induced aldoxime formation in western balsam poplar (Populus trichocarpa). Heterologous expression in Saccharomyces cerevisiae revealed that both enzymes produce a mixture of different aldoximes. Knockdown lines of CYP79D6/7 in gray poplar (Populus 3 canescens) exhibited a decreased emission of aldoximes, nitriles, and alcohols, emphasizing that the CYP79s catalyze the first step in the formation of a complex volatile blend. Aldoxime emission was found to be restricted to herbivore-damaged leaves and is closely correlated with CYP79D6 and CYP79D7 gene expression. The semi-volatile phenylacetaldoxime decreased survival and weight gain of gypsy moth (Lymantria dispar) caterpillars, suggesting that aldoximes may be involved in direct defense. The wide distribution of volatile aldoximes throughout the plant kingdom and the presence of CYP79 genes in all sequenced genomes of angiosperms suggest that volatile formation mediated by CYP79s is a general phenomenon in the plant kingdom.

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“…The sample is introduced either by manual direct infusion (DI) or via HPLC or UHPLC followed by DI, following which the sample is delivered to the probe via a capillary; the probe functions to translocate the liquid sample from the capillary to the needle of the ion source. Two ionisation sources are generally employed in LC-MS based metabolomics: electrospray ionisation (ESI) is the most commonly employed (Tolstikov et al, 2003;Jander et al, 2004;von Roepenack-Lahaye et al, 2004;Vorst et al, 2005;Moco et al, 2006;Rischer et al, 2006;De Vos et al, 2007;Hanhineva et al, 2008) and is particularly well suited to the ionisation of a wide range of metabolites including, drug compounds (Chen et al, 2007), amino and organic acids Figure 3. The relationship between column stationary phase particle size, fl ow velocity and the height equivalent to the theoretical plate (HETP) value as an expression of chromatographic separation effi ciency.…”

Section: Sample Introduction and Ionisation Methods Employed In Liquimentioning

confidence: 99%

“…chromatography combined with mass spectrometry (MS) or nuclear magnetic resonance (NMR) spectroscopy is capable of detecting all metabolites within a given biological sample Hall, 2006;Allwood et al, 2008). Over the past decade many methods for the high-throughput metabolomic analysis of plant derived samples have been established for gas chromatography combined with MS (GC-MS; Fiehn et al, 2000;Fernie et al, 2004;Lisec et al, 2006), direct injection mass spectrometry (DIMS; Goodacre et al, 2003;Catchpole et al, 2005;Allwood et al, 2006), liquid chromatography MS (LC-MS; Tolstikov et al, 2003;Jander et al, 2004;von Roepenack-Lahaye et al, 2004;Vorst et al, 2005;Moco et al, 2006;Rischer et al, 2006;De Vos et al, 2007), capillary electrophoresis MS (CE-MS; Sato et al, 2004), and 1 H-NMR (Ratcliff e and Shachar-Hill, 2001;Le Gall et al, 2003;Ward et al, 2003;Choi et al, 2004Choi et al, , 2006. 1 H-NMR has proven to be an appropriate tool for untargeted plant metabolomics, especially where studies focus upon samples that contain highly abundant bulk metabolite species, for example sugars in fruits (Biais et al, 2009).…”

mentioning

confidence: 99%

An introduction to liquid chromatography–mass spectrometry instrumentation applied in plant metabolomic analyses

Allwood

Goodacre

2009

Phytochemical Analysis

218

143

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Over the past decade the application of non-targeted high-throughput metabolomic analysis within the plant sciences has gained ever increasing interest and has truly established itself as a valuable tool for plant functional genomics and studies of plant biochemical composition. Whilst proton nuclear magnetic resonance ( 1 H-NMR) spectroscopy is particularly appropriate for the analysis of bulk metabolites and gas chromatography mass spectrometry (GC-MS) to the analysis of volatile organic compounds (VOC's) and derivatised primary metabolites, liquid chromatography (LC)-MS is highly applicable to the analysis of a wide range of semi-polar compounds including many secondary metabolites of interest to plant researchers and nutritionists. In view of the recent developments in the separation sciences, leading to the advent of ultra high performance liquid chromatography (UHPLC) and MS based technology showing the ever improving resolution of metabolite species and precision of mass measurements (sub-ppm accuracy now being achievable), this review sets out to introduce the background and update the reader upon LC, high performance (HP)LC and UHPLC, as well as the large range of MS instruments that are being applied in current plant metabolomic studies. As well as covering the theory behind modern day LC-MS, the review also discusses the most relevant metabolomics applications for the wide range of MS instruments that are currently being applied to LC.

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Application of a high‐throughput HPLC‐MS/MS assay to Arabidopsis mutant screening; evidence that threonine aldolase plays a role in seed nutritional quality

Cited by 122 publications

References 44 publications

Metabolic cross-feeding via intercellular nanotubes among bacteria

Metabolic cross-feeding via intercellular nanotubes among bacteria

Two Herbivore-Induced Cytochrome P450 Enzymes CYP79D6 and CYP79D7 Catalyze the Formation of Volatile Aldoximes Involved in Poplar Defense

An introduction to liquid chromatography–mass spectrometry instrumentation applied in plant metabolomic analyses

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