Abstract
Estimation of stomatal aperture using low viscosity silicone-base impression material has the advantage of working with the whole leaf. The developmental stage and the environment strongly affect the stomatal aperture. Therefore, it is mandatory to have accurate estimations of the stomatal aperture of intact leaves under different situations. With this technique, it is possible to get the real picture at any moment. The outputs of the data include studies on cell area and morphology, epidermis cell and stomata lineages, among others. This protocol is useful for the accurate estimation of stomatal aperture in many samples of intact leaves in Arabidopsis thaliana.
Keywords: Drought, Epidermis, Leaf development, Oxidative stress, Photorespiration, Stomatal density
Background
The epidermis of all leaves has specialized cells, the guard cells, surrounding microscopic pores. The guard cells and pores are called stomata, and they permit gas exchange and diffusion of water vapor between the atmosphere and the interior of the leaf. Stomata are products of an intracellular program, which generates the specific stomatal patterns during their development (Kagan et al., 1992). Stomata are found both in the abaxial and adaxial sides of the leaf, although the stomatal density (and starch accumulation) is higher on the abaxial side of the leaf sheath in Arabidopsis thaliana (Schlüler et al., 2003; Tsai et al., 2009). The stomatal density is controlled by endogenous and exogenous factors in Arabidopsis thaliana (Berger and Altmann, 2000; von Groll et al., 2002). Stomatal aperture actively responds to changes in the environment and regulates leaf transpiration rates (Santelia and Lawson, 2016). An accurate estimation of the stomatal aperture provides insight into the impact of environmental stress on plants.
Materials and Reagents
Equipment
Software
Procedure
Data analysis
The number of stomata analyzed on a typical silicon imprint was 30-40. These stomata were randomly selected in the separate microscopic fields in the same slide. Ten leaves per Arabidopsis line were analyzed.
Note: Other results of stomatal aperture were published in Scarpeci et al., 2017.
Acknowledgments
This work was supported by ANPCyT and CONICET, Argentina. This protocol was modified from previous work (Weyers and Johansen, 1985).
References
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