Fast and Simple Droplet Sampling of Sap from Plant Tissues and Capillary Microextraction of Soluble Saccharides for Picogram-Scale Quantitative Determination with GC-MS

Tan, Yupeng; Li, Ke; Hu, Lei; Chen, Shu; Gai, Ying; Jiang, Xiangning

doi:10.1021/jf102053h

Cited by 13 publications

(10 citation statements)

References 11 publications

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“…More recently, Malagon et al and Zhang et al detected rhamnitol in human urine using GC/MS (Malagon et al, 2006) and LC/MS with HILIC (Zhang, Creek, Barrett, Blackburn, & Watson, 2012). Rhamnitol was detected in the leaves of Arabidopsis thaliana, corn grains (Zea maize), and the fruit of cucumber (Cucumis sativus) by GC/MS; however, it was not found in Fuji apples, in contrast to our results (Li et al, 2013;Tan et al, 2010). In the only study of fruits, Yun et al identified rhamnitol in Satsuma mandarin (Citrus unshiu Marcovitch) (Yun et al, 2013).…”

Section: Apple Characterisation By Targeted Analysis With Annotated Mcontrasting

confidence: 86%

A NMR-based, non-targeted multistep metabolic profiling revealed l-rhamnitol as a metabolite that characterised apples from different geographic origins

et al. 2015

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Section: Apple Characterisation By Targeted Analysis With Annotated Mcontrasting

confidence: 86%

A NMR-based, non-targeted multistep metabolic profiling revealed l-rhamnitol as a metabolite that characterised apples from different geographic origins

et al. 2015

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“…Levels of seed soluble monosaccharides (sucrose, glucose and fructose) were measured by gas chromatography mass spectrometry (GC–MS) analysis as previously reported (Tan et al 2010 ; Li et al 2013 ) with some modifications. Briefly, 200 mg flour samples were dissolved in 2 mL Me 2 SO, mixed with 150 μL of acetic anhydride and 30 μL of 1-methylimidazole, and stirred for 10 min in glass tubes; 600 μL of double-distilled H 2 O (ddH 2 O) was added to each tube to remove excess acetic anhydride, and 100 μL of CH 2 Cl 2 was added to extract the acetylated derivatives.…”

Section: Methodsmentioning

confidence: 99%

Pyrophosphate: fructose-6-phosphate 1-phosphotransferase (PFP) regulates carbon metabolism during grain filling in rice

et al. 2016

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Key message Decreased PFPase activity in rice perturbs the equilibration of carbon metabolism during grain filling but has no visible phenotypic effects during the vegetative and reproductive growth stages. Abstract Starch is a primary energy reserve for various metabolic processes in plant. Despite much advance has been achieved in pathways involved in starch biosynthesis, information was still lacked for precise regulation related to carbon metabolism during seed filling in rice (Oryza sativa). The objective of this study was to identify and characterize new gene associated with carbon metabolism during grain filling. By screening our chemical mutant pool, two allelic mutants exhibiting floury endosperm were isolated. No visible phenotypic defects were observed during both the vegetative and reproductive growth stages, except for the floury-like endosperm of grains with significantly reduced kernel thickness, 1000-grain weight and total starch content. Map-based cloning revealed that the mutant phenotypes were controlled by a gene encoding pyrophosphate: fructose-6-phosphate 1-phosphotransferase (PFP, EC 2.7.1.90) b subunit (PFP b ), which catalyzes reversible interconversion between fructose-6-phosphate and fructose-1, 6-bisphosphate. The identity of PFP b was further confirmed by a genetic complementation test. Subcellular analysis demonstrated that PFPb was localized in cytoplasm. Quantitative PCR and histochemical staining indicated PFP b was ubiquitously expressed in various tissues. Furthermore, we found PFP b could express in both the early and late phases of starch accumulation during grain filling and decreased activity of PFP b in pfp mutants resulted in compromised carbon metabolism with increased soluble sugar contents and unfavorable starch biosynthesis. Our results highlight PFP b functions in modulating carbon metabolism during grain filling stage.

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“…Glucose, fructose, galactose, and sucrose in flag leaf blade, internode and panicle of WT and mutant plants were extracted with 80% methanol and quantified by GC-MS analysis as previously described (Tan et al 2010).…”

Section: Determination Of Sugar Contentsmentioning

confidence: 99%

Downregulation of OsPK1, a cytosolic pyruvate kinase, by T-DNA insertion causes dwarfism and panicle enclosure in rice

Zhang

Xiao

Luo

et al. 2011

Planta

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Pyruvate kinase (PK) catalyzes the final step of glycolysis. There are few reports on the role of PK in rice. Here, we identified a novel rice dwarf mutant, designated as ospk1, showing dwarfism, panicle enclosure, reduced seed set, and outgrowth of axillary buds from culm nodes. Sequence analyses of 5'-RACE indicated that a single T-DNA was inserted in the transcriptional regulatory region of OsPK1 in ospk1. Quantitative RT-PCR result showed that OsPK1 expression was decreased by approximately 90% in ospk1 compared with that in WT. Enzyme assay and transient expression in protoplasts indicated that OsPK1 encodes a cytosolic PK (PK(c)). Complementation with OsPK1 demonstrated that OsPK1 is responsible for the phenotype of ospk1. Quantitative RT-PCR and GUS staining analyses exhibited that OsPK1 was expressed mainly in leaf mesophyll cells, phloem companion cells in stems, and cortical parenchyma cells in roots. The transcriptions of four other putative enzymes involved in the glycolysis/gluconeogenesis pathway were altered in ospk1. The amount of pyruvate is decreased in ospk1. We propose that OsPK1 plays an important role through affecting the glycolytic pathway. The contents of glucose and fructose were markedly accumulated in flag leaf blade and panicle of ospk1. The sucrose level in panicle of ospk1 was decreased by approximately 84%. These findings indicated that both monosaccharide metabolism and sugar transport are altered due to the decreased expression of OsPK1. Together, these results provide new insights into the role of PK(c) in plant morphological development, especially plant height.

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Fast and Simple Droplet Sampling of Sap from Plant Tissues and Capillary Microextraction of Soluble Saccharides for Picogram-Scale Quantitative Determination with GC-MS

Cited by 13 publications

References 11 publications

A NMR-based, non-targeted multistep metabolic profiling revealed l-rhamnitol as a metabolite that characterised apples from different geographic origins

A NMR-based, non-targeted multistep metabolic profiling revealed l-rhamnitol as a metabolite that characterised apples from different geographic origins

Pyrophosphate: fructose-6-phosphate 1-phosphotransferase (PFP) regulates carbon metabolism during grain filling in rice

Downregulation of OsPK1, a cytosolic pyruvate kinase, by T-DNA insertion causes dwarfism and panicle enclosure in rice

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