Leaf Stomatal Traits Measurement

Samples from the middle portion of leaf blades (avoiding the midrib) were obtained, and then stored and processed for posterior analyses exactly as described elsewhere [67 (link)]. For each sample, five fields of view were used for measuring SD, SI, and SS. Magnifications were 10× for SD, and 20× for SI and SS. The SD was computed as the total number of stomata per area, and SI was calculated as the stomatal number-to-epidermal cell number ratio. The SS was calculated as guard cell length multiplied by the width of the guard cell pair according to Franks & Beerling [68 (link)]. The slides were photographed using a digital camera (Zeis AxioCan HRc, Göttingen, Germany) mounted on a light microscope (AX70 TRF, Olympus Optical, Tokyo, Japan); images were captured and analyzed using the Image-Pro Plus 4.5 software (Media Cybernetics, Silver Spring, ML, USA). Additionally, g_wmax was calculated based on anatomical traits, exactly as described in [12 (link)].

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de Souza A.H., de Oliveira U.S., Oliveira L.A., de Carvalho P.H., de Andrade M.T., Pereira T.S., Gomes Junior C.C., Cardoso A.A., Ramalho J.D., Martins S.C, & DaMatta F.M. (2023). Growth and Leaf Gas Exchange Upregulation by Elevated [CO2] Is Light Dependent in Coffee Plants. Plants, 12(7), 1479.

Publication 2023

AX70 TRF Image-Pro Plus 4.5

Camera Cell Epidermal cell Leaf Microscope light Olympus Silver Stomatal

Corresponding Organization : Universidade Federal de Viçosa

Other organizations : North Carolina State University, University of Lisbon, Universidade Nova de Lisboa

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Stomatal density (SD)
Stomatal index (SI)
Stomatal size (SS)
Maximum stomatal conductance (g_wmax)

control variables

Sampling location (middle portion of leaf blades, avoiding the midrib)
Sample storage and processing
Microscope magnification (10x for SD, 20x for SI and SS)
Imaging software (Image-Pro Plus 4.5)

controls

None explicitly mentioned

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