Comprehensive multiphase NMR: a promising technology to study plants in their native state

Abstract

Nuclear magnetic resonance (NMR) spectroscopy is arguably one the most powerful tools to study the interactions and molecular structure within plants. Traditionally, however, NMR has developed as two separate fields, one dealing with liquids and the other dealing with solids. Plants in their native state contain components that are soluble, swollen, and true solids. Here, a new form of NMR spectroscopy, developed in 2012, termed comprehensive multiphase (CMP)-NMR is applied for plant analysis. The technology composes all aspects of solution, gel, and solid-state NMR into a single NMR probe such that all components in all phases in native unaltered samples can be studied and differentiated in situ. The technology is evaluated using wild-type Arabidopsis thaliana and the cellulose-deficient mutant ectopic lignification1 (eli1) as examples. Using CMP-NMR to study intact samples eliminated the bias introduced by extraction methods and enabled the acquisition of a more complete structural and metabolic profile; thus, CMP-NMR revealed molecular differences between wild type (WT) and eli1 that could be overlooked by conventional methods. Methanol, fatty acids and/or lipids, glutamine, phenylalanine, starch, and nucleic acids were more abundant in eli1 than in WT. Pentaglycine was present in A. thaliana seedlings and more abundant in eli1 than in WT.

Focusing on “the important” through targeted NMR experiments: an example of selective13C–12C bond detection in complex mixtures

“…Working in Reverse: 13 C 6 -Glucose in Arabidopsis thaliana Figure 5 depicts the germination and early growth of 12 C Arabidopsis thaliana seeds in the dark using 13 C 6 -Glucose as the sole carbon source as previously described. 61 This study has been designed as the reverse of the Daphnia study such that the process begins with 12 C carbon, and 13 C is added into the system. As the plants are stored in the dark, photosynthesis does not occur, and the seeds grow via 13 C sorption through its husk and later uptake via roots after initial germination.…”

In-Vivo NMR Spectroscopy: A Powerful and Complimentary Tool for Understanding Environmental Toxicity

“…However, since 2001 there has been an interest in environmental studies as a method to detect and explain toxicity [ 23 , 30 ]. Environmental metabolomics studies changes in the metabolite profile with changing environmental conditions [ 30 ], and have been used on a host of different species including: worms [ 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 ], Drosophila melanogaster [ 39 ], Daphnia magna [ 20 , 40 , 41 , 42 , 43 , 44 , 45 ], Hyalella azteca [ 46 , 47 , 48 ], Caenorhabditis elegans [ 49 , 50 , 51 ], rodents [ 52 , 53 , 54 , 55 ], and plants [ 56 , 57 , 58 , 59 , 60 ]. Species are often chosen either due to their susceptibility to external stressors or their abundance geographically.…”

In vivo comprehensive multiphase NMR

“…In many ways, CMP NMR can be thought of as changing NMR technology to match natural samples, rather than changing the sample to match a specific NMR technique. CMP NMR has permitted the study of structural interactions and processes in unaltered heterogeneous samples, including soil, swelling of rubber by biofuels, intact plants, and to follow biological processes (seed growth) …”