Abstract
Arabidopsis thaliana is the most widely used model organism for research in plant biology. While significant advances in understanding plant growth and development have been made by focusing on the molecular genetics of Arabidopsis, extracting and understanding the functional framework of metabolism is challenging, both from a technical perspective due to losses and modification during extraction of metabolites from the leaves, and from the biological perspective, due to random variation obscuring how well the function is performed. The purpose of this work is to establish the in vivo metabolic profile directly from the Arabidopsis thaliana leaves without metabolite extraction, to reduce the complexity of the results by multivariate analysis, and to unravel the mitigation of cellular complexity by predominant functional periodicity. To achieve this, we use the circadian cycle that strongly influences metabolic and physiological processes and exerts control over the photosynthetic machinery. High resolution-magic angle spinning nuclear magnetic resonance (HR-MAS NMR) was applied to obtain the metabolic profile directly from intact Arabidopsis leaves. Combining one- and two-dimensional 1H HR-MAS NMR allowed the identification of several metabolites including sugars and amino acids in intact leaves. Multivariate analysis on HR-MAS NMR spectra of leaves throughout the circadian cycle revealed modules of primary metabolites with significant and consistent variations of their molecular components at different time points of the circadian cycle. Since robust photosynthetic performance in plants relies on the functional periodicity of the circadian rhythm, our results show that HR-MAS NMR promises to be an important non-invasive method that can be used for metabolomics of the Arabidopsis thaliana mutants with altered physiology and photosynthetic efficiency.
Highlights
As a model organism, Arabidopsis thaliana play a central role in understanding biological functions across plant species and in characterizing phenotypes associated with genetic mutations [1]
Our results demonstrate that High resolution magic angle spinning (HR-magic angle spinning (MAS)) NMR on intact Arabidopsis leaves represents a novel platform that could provide important in vivo information of regular metabolic network against which altered metabolic profile due to stress, infection or mutation can be assessed
Peak assignment was performed according to earlier literature and the Biological Magnetic Resonance Data Bank (BMRB) [25,26]
Summary
Arabidopsis thaliana play a central role in understanding biological functions across plant species and in characterizing phenotypes associated with genetic mutations [1]. Significant advances in understanding plant growth and development have been made by focusing on molecular genetics of Arabidopsis. Understanding the functional framework of metabolism in native state in leaves poses a major challenge for all metabolomics approaches. Many approaches, including mass spectrometry as well as NMR methods require labour-intensive extraction of plant metabolites which can cause biases resulting from differential extraction efficiencies and from the loss of volatile metabolites [4,5]. Extraction methods cause the loss of molecular information regarding specific associations within and between polymeric structural plant components
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