Trehalose determination in Norway spruce (Picea abies) roots. Analytics matters
Miloslav Šulc,
Ivana Tomášková,
Anna Krejzková,
Michal Samek,
Alina Diuzheva,
Jaromír Hradecký,
Vítězslava Pešková
Affiliations
Miloslav Šulc
Food Research Institute, Radiova 1285/7, CZ-10200 Prague 15 - Hostivar, Czechia; Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129- Suchdol, CZ - 165 21 Praha 6, Czechia; Corresponding author at: Czech University of Life Sciences Prague, Faculty of Forestry and Wood Sciences, Kamýcká 129, CZ - 165 21 Praha 6 - Suchdol, Czech Republic.
Ivana Tomášková
Faculty of Forestry and Wood Sciences, Czech University of Life Sciences Prague, Kamýcká 129 Suchdol, CZ - 165 21 Praha 6, Czechia
Anna Krejzková
Faculty of Forestry and Wood Sciences, Czech University of Life Sciences Prague, Kamýcká 129 Suchdol, CZ - 165 21 Praha 6, Czechia
Michal Samek
Faculty of Forestry and Wood Sciences, Czech University of Life Sciences Prague, Kamýcká 129 Suchdol, CZ - 165 21 Praha 6, Czechia
Alina Diuzheva
Faculty of Forestry and Wood Sciences, Czech University of Life Sciences Prague, Kamýcká 129 Suchdol, CZ - 165 21 Praha 6, Czechia
Jaromír Hradecký
Faculty of Forestry and Wood Sciences, Czech University of Life Sciences Prague, Kamýcká 129 Suchdol, CZ - 165 21 Praha 6, Czechia
Vítězslava Pešková
Faculty of Forestry and Wood Sciences, Czech University of Life Sciences Prague, Kamýcká 129 Suchdol, CZ - 165 21 Praha 6, Czechia
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.
