The stomatal pore condition and xylem structure were analyzed using an Olympus BX50 microscope (Olympus, Tokyo, Japan). The preparation of leaf samples for stomatal analysis was described previously in [18 (link)]. Fresh leaflets were collected from plants under control and water-withholding stress conditions, and a thick tape was pasted on their upper surface. The tape was gently pulled from the leaflet to tear off the epidermis layer and placed on a glass slide. Other leaf parts were cut off using a scalpel. A coverslip was placed on the sample after adding a drop of water. The slides were observed at 1000× magnification to determine the stomatal pore area. Due to their elliptical shape, the following formula was used to calculate the stomatal pore area:
where π = 3.14, and r1 and r2 are the minor and major radii of the stomatal pores, respectively.
To observe the xylem structure, a cross section of the stem was obtained from one-month-old plants under control and drought stress using a scalpel. Then, the sections were immersed in 0.05% toluidine-blue-O (TBO) (Waldeck, Münster, German) for 30 s and washed with dH2O several times. The sections were then examined under 100× A magnification using an Olympus BX50 microscope (Olympus, Tokyo, Japan).
where π = 3.14, and r1 and r2 are the minor and major radii of the stomatal pores, respectively.
To observe the xylem structure, a cross section of the stem was obtained from one-month-old plants under control and drought stress using a scalpel. Then, the sections were immersed in 0.05% toluidine-blue-O (TBO) (Waldeck, Münster, German) for 30 s and washed with dH2O several times. The sections were then examined under 100× A magnification using an Olympus BX50 microscope (Olympus, Tokyo, Japan).
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