Development of a root exudate collection protocol for metabolomics analysis using Nuclear Magnetic Resonance

Fortier, Mélanie; Lemyre, Julie; Ancelin, Edouard; Oulyadi, Hassan; Driouich, Azeddine; Vicré, Maïté; Follet-Gueye, Marie-Laure; Guilhaudis, Laure

doi:10.1016/j.plantsci.2023.111694

Cited by 5 publications

(1 citation statement)

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“…Sensitivity and separation capacities of the analytical methods achieved in the last decade, enable a better screening of the diversity of the chemical signals released by plants or microbiome [13]. However, most of these studies focused on root exudates obtained from plants grown hydroponically or on gelose with a nutritive solution [14][15][16][17][18], meaning roots lacked living substrate as is the soil. On the other hand, several studies performed in microcosms tested how the presence of soil fauna may contribute to mineralization but mainly focused on analyzing C, N, or P content of the leachates [19][20][21][22].…”

Section: Introductionmentioning

confidence: 99%

Studying chemical signals in plant–soil mesofauna interactions

Bohm,

Michel,

Forey

et al. 2024

Comptes Rendus. Chimie

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Discrimination of compounds produced from plant roots is a challenging purpose, especially in presence of the soil matrix and the associated fauna. In this study, we aimed to test a collection method of chemical signals released by living plants and soil fauna in interaction in a soil matrix. Microcosms, with stopcock, were filled with soil, and four treatments were designed based on the presence/absence of an annual herbaceous plant, Poa annua and of a soil Collembola species, Folsomia candida. Soil leachates were collected with a percolated solution and analyzed by a fast analytical method combining analytical separation of compounds through ultra high performance liquid chromatography with high resolution mass spectrometry (UHPLC-HRMS). The experimental design appears to be suitable for leachates sampling. Detected chemicals clearly discriminated the treatments, demonstrating how biotic interactions belowground between a plant and a Collembola species may change soil chemical signals.

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Section: Introductionmentioning

confidence: 99%

Studying chemical signals in plant–soil mesofauna interactions

Bohm,

Michel,

Forey

et al. 2024

Comptes Rendus. Chimie

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Targeting the untargeted: Uncovering the chemical complexity of root exudates

Möller,

Ritter,

Stobinsky

et al. 2024

Preprint

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The chemical complexity of root exudates has garnered significant attention in recent years, yet critical gaps remain in understanding the full scope of root exudate chemical variation across the plant kingdom. To address this, we conducted a systematic review of 57 studies, comprising 124 experiments, aimed at evaluating current methodologies and findings in untargeted root exudate chemical analysis. Our review revealed that hydroponic (44%) and soil-hydroponic hybrid (32%) sampling approaches, primarily utilising water as the collection medium, were the most common experimental setups. Liquid chromatography-mass spectrometry (LC-MS) was the predominant analytical technique used in 54% of the studies, followed by gas chromatography-mass spectrometry (GC-MS) in 31%. The average number of metabolites identified per analysis was 960, though the number of annotated metabolites varied considerably. Shikimates, phenylpropanoids, and carbohydrates were the most frequently identified classes, with their relative abundances varying widely. Several methodological challenges were highlighted, including inconsistencies in sampling techniques, underrepresentation of non-crop plants, and incomplete chemical annotation. To address these limitations, we propose a framework emphasising the need for representative exudate sampling, the use of multiple analytical approaches, the development of advanced bioinformatics tools, and the integration of these findings to enhance our understanding of root exudates and their ecological functions.

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Root Exudation: An In‐Depth Experimental Guide

McLaughlin,

Himmighofen,

Khan

et al. 2024

Plant Cell & Environment

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Plants exude a wide variety of compounds into the rhizosphere, modulating soil functioning and diversity. The number of studies investigating exudation has exponentially increased over the past decades. Yet, the high inter‐study variability of the results is slowing down our understanding of root–soil interactions. This variability is partly due to the absence of harmonized methodologies to collect and characterize exudation. Here, we discuss how various experimental aspects influence exudation profiles by performing a literature review, and we suggest best practices for different experimental setups. We discuss state‐of‐the‐art of spatially resolved exudate collection, collection in controlled versus field conditions and plant growth setups ranging from hydroponics to soil. We highlight the importance of preparing experimental blanks, in situ versus ex situ exudate collection, various collection media and timing of collection, exudate storage and processing and analytical considerations. We summarize best practices for experimental setup and reporting of parameters in an easily accessible table format to facilitate discussion of best practices in the field. An increased standardization in the field together with the systematic studies suggested will improve our knowledge of how plant exudation shapes interactions with organisms in soil.

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Development of a root exudate collection protocol for metabolomics analysis using Nuclear Magnetic Resonance

Cited by 5 publications

References 62 publications

Studying chemical signals in plant–soil mesofauna interactions

Studying chemical signals in plant–soil mesofauna interactions

Targeting the untargeted: Uncovering the chemical complexity of root exudates

Root Exudation: An In‐Depth Experimental Guide

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