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.
Physiol Plant. 2013 Mar;147(3):261-9. doi: 10.1111/j.1399-3054.2012.01677.x. Epub 2012 Sep 13.
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.
海藻糖是一种非还原性二糖，参与植物的应激耐受。为了更好地理解海藻糖在亚麻（Linum usitatissimum）渗透胁迫响应中的作用，研究了叶片中海藻糖的含量。首先，常用的糖测定方法——高效阴离子交换色谱-脉冲安培检测法（HPAEC-PAD）的结果并不令人满意，而且通过改变洗脱条件也无法提高分离效率。同样的问题也出现在模式植物拟南芥中。通过海藻糖酶测定和液相色谱-高分辨质谱分析清楚地表明，这两种植物中海藻糖存在共洗脱后，改用气相色谱-质谱法（GC-MS）作为分析方法。这些结果证实，HPAEC-PAD 分析目前高估了海藻糖的含量，拟南芥和亚麻的含量分别约为 7 倍和 13 倍。因此，GC-MS 更能满足植物中海藻糖定量分析的需要。使用该方法，在渗透胁迫（-0.30 MPa）下观察到亚麻中海藻糖的积累，在 48 小时后其含量（31.49 nmol g-1 干重）太低，无法将其赋予应激亚麻中的海藻糖作为渗透保护剂或渗透调节剂的作用。
