Perforated-tape Epidermal Detachment (PED): A simple and rapid method for isolating epidermal peels from specific areas of Arabidopsis leaves
Perforated-tape Epidermal Detachment (PED): A simple and rapid method for isolating epidermal peels from specific areas of Arabidopsis leaves
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DOI：
10.5511/plantbiotechnology.13.0903b
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发表时间：
2013-01-01
影响因子：
1.6
通讯作者：
Mochizuki, Nobuyoshi
中科院分区：
文献类型：
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作者：
Ibata, Harue;Nagatani, Akira;Mochizuki, Nobuyoshi
叶表皮气孔在高等植物的气体交换和蒸腾作用中起着关键作用。气孔开闭（气孔运动）和气孔形态发生的调控机制已得到深入研究。此类研究需要收集分离的表皮细胞层（“皮”），以便在显微镜下详细观察气孔。然而，用于制备和处理表皮剥落物的常规技术存在缺点，即需要一定水平的技能或设备(例如，韦林氏搅拌机)。在本报告中，我们提出了一种简单快速的表皮剥离方法，称为穿孔胶带表皮脱离（PED）。对于PED，将时间胶带粘附到拟南芥叶的近轴表皮上，并使用打有小孔的透明胶带将远轴表皮层分离。孔内的区域适合显微镜观察，因为透明胶带不会掩盖那里分离的表皮层。 PED 还可用于制备烟草和豆类叶的表皮。与传统混合法相比，PED 的关键优势在于，PED 制备的表皮细胞很少受到损伤，并且我们证明 PED 可用于获得用于气孔对蓝光和外部应用 ABA 反应的生理测定的材料。因此，我们认为 PED 方法适合制备用于生理研究的表皮细胞层。
Leaf epidermal stomata play pivotal roles in gas exchange and transpiration in higher plants. The regulatory mechanisms of the opening and closing of stomata (stomatal movement) and stomatal morphogenesis have been intensively studied. Such studies require the collection of detached epidermal cell layers ("peels") for the detailed observation of stomata under the microscope. However, there are drawbacks to the conventional techniques for preparing and handling epidermal peels, i.e., a certain level of skill or an apparatus (e. g., a Waring blender) is required. In this report, we present a simple and rapid method for preparing epidermal peels called Perforated-tape Epidermal Detachment (PED). For PED, Time Tape is adhered to the adaxial epidermis of an Arabidopsis leaf, and the abaxial epidermal layer is detached using Scotch Tape perforated with a small hole. The area inside the hole is suitable for microscopic observation because the Scotch Tape does not mask the detached epidermal layer there. PED can also be used to prepare epidermal peels from tobacco and legume leaves. The crucial advantage of PED over the conventional blender method is that the epidermal cells prepared by PED are rarely damaged, and we demonstrate that PED can be used to obtain material for physiological assays of stomatal responses to blue light and externally applied ABA. We thus believe that the PED method is suitable for preparing epidermal cell layers for physiological studies.