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. 2021 Feb 18:8:101280.
doi: 10.1016/j.mex.2021.101280. eCollection 2021.

Trehalose determination in Norway spruce (Picea abies) roots. Analytics matters

Miloslav Šulc  1   2 , Ivana Tomášková  3 , Anna Krejzková  3 , Michal Samek  3 , Alina Diuzheva  3 , Jaromír Hradecký  3 , Vítězslava Pešková  3
Affiliations

Trehalose determination in Norway spruce (Picea abies) roots. Analytics matters

Miloslav Šulc et al. MethodsX. .

Abstract

We present concise results of method validation for trehalose quantitation by LC-MS/MS in spruce ectomycorrhizal roots in order to describe spruce health status, mainly in connection to contamination by a pathogenic fungus, Gemmamyces piceae. The procedure is based on Rogatsky et al. (2005) developed for human plasma. We found out that the best extraction yield was achieved with 80% methanol/water (v/v) solution and optimal extraction temperature was set between 50-60°C. In contrast to previous papers, we minimized the activity of trehalase enzyme by putting root samples into liquid N2 immediately after root excavation, followed by freeze-drying in order to stop trehalase activity. Higher content of trehalose was recorded in healthy trees, confirming the idea that ectomycorrhiza plays a significant role in plant-pathogen interactions.

Keywords: Liquid chromatography - Mass spectrometry; Mycorrhiza; Spruce roots; Trehalose.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper.

Figures

Image, graphical abstract
Graphical abstract
Fig 1
Fig. 1
Chromatogram comparison from a freeze-dried material of a healthy (red) and diseased (blue) tree. Retention time of trehalose is about 4.5 min.
See this image and copyright information in PMC

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