Trehalose determination in linseed subjected to osmotic stress. HPAEC-PAD analysis: an inappropriate method.
Trehalose is a non-reducing disaccharide involved in stress tolerance in plants. To understand better the role of trehalose in the osmotic stress response in linseed (Linum usitatissimum), trehalose content in leaves was studied. First, the method commonly used for sugar determination, high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD), gave unsatisfactory results and the separation efficiency could not be improved by varying the elution conditions. The same problem was also found in the model plant: Arabidopsis thaliana. After clearly highlighting a co-elution of trehalose in these two species by a trehalase assay and liquid chromatography-high resolution mass spectrometry analysis, gas chromatography-mass spectrometry (GC-MS) was used as the analytical method instead. These results confirmed that trehalose content is currently overestimated by HPAEC-PAD analysis, approximately 7 and 13 times for A. thaliana and linseed respectively. Thus GC-MS gave more satisfactory results for trehalose quantification in plants. With this method, trehalose accumulation was observed in linseed during an osmotic stress (-0.30 MPa), the quantity (31.49 nmol g(-1)  dry weight after 48 h) appears too low to assign an osmoprotector or osmoregulator role to trehalose in stressed linseed.
Département de Génie Biologique, Université de Picardie Jules Verne, BIOPI EA3900 Biologie des Plantes & Innovation - LPMV, IUT d'Amiens, Avenue des Facultés, 80025, Amiens Cedex, France.
Anthony
A
Quéro
Linda
L
Béthencourt
Serge
S
Pilard
Antoine
A
Fournet
Xavier
X
Guillot
Rajbir S
RS
Sangwan
Michèle
M
Boitel-Conti
Josiane
J
Courtois
Emmanuel
E
Petit
D017360:Arabidopsis / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D019597:Flax / Q000378:metabolism* / Q000502:physiology; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D005947:Glucose / Q000032:analysis / Q000378:metabolism; D059625:Liquid-Liquid Extraction; D009995:Osmosis; D010936:Plant Extracts / Q000032:analysis; D018515:Plant Leaves / Q000378:metabolism; D014198:Trehalase / Q000378:metabolism; D014199:Trehalose / Q000032:analysis* / Q000378:metabolism
D010936:Plant Extracts; D014199:Trehalose; D014198:Trehalase; D005947:Glucose