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Fig 1.

Stomatal aperture response to variations in pCO₂.

Stomatal aperture (SA; n = 800) was measured in plant taxa from different altitudinal origins: high-altitude taxa (“high”; altitude of origin 2,970 m a.s.l.), including Anthyllis vulneraria subsp. valesiaca and Arabis alpina, and low-altitude taxa (“low”; 540 m a.s.l.), including Anthyllis vulneraria subsp. carpatica and Arabidopsis thaliana (Col-0). All plants were cultivated under reduced (30 Pa) and ambient pCO₂ (42 Pa). Marginal means along with their 95% confidence intervals are presented. Statistical analysis results are provided in S3 Table. Different letters denote statistically significant differences at p < 0.05, as determined by post hoc tests.

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Fig 1 Expand

Table 1.

Marginal effects of environmental factors on stomatal aperture.

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Table 1 Expand

Fig 2.

Marginal effects of environmental factors on stomatal aperture.

Effects of (A) partial pressure of CO2 (pCO₂), (B) air temperature, (C) vapor pressure deficit (VPD), and (D) irradiance on stomatal aperture (SA; n = 1’200) of Arabis alpina and Arabidopsis thaliana (Col-0). The data were obtained from four experiments conducted under environmental conditions that varied within the following minimum and maximum ranges: pCO₂ (29.1–51.8 Pa), temperature (18.8–21.6°C), VPD (7.7–13.2 hPa), and irradiance (85 and 175 µmol m ⁻ ² s ⁻ ¹). Marginal means along with their 95% confidence intervals are presented. Statistical analysis results are provided in Table 1.

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Fig 3.

Stomatal frequency of plants growing at their natural sites.

(A) Stomatal density (SD; n = 82) and (B) stomatal index (SI; n = 82) were measured in plants growing at their natural sites. High-altitude taxa (“high”; 2,970 m a.s.l., pCO₂ approx. 29 Pa in 2018, the year of sampling) included Anthyllis vulneraria subsp. valesiaca and Arabis alpina, whereas low-altitude taxa (“low”; 540 m a.s.l., pCO₂ approx. 39 Pa) comprised Anthyllis vulneraria subsp. carpatica and Arabidopsis thaliana. Marginal means along with their 95% confidence intervals are presented. Statistical analysis results are provided in S4 Table. Different letters denote statistically significant differences at p < 0.05, as determined by post hoc tests.

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Fig 4.

Stomatal frequency of plants from natural sites and their laboratory-grown conspecifics exposed to altitude-specific pCO2 levels.

Stomatal density (SD; A, n = 262; C, n = 239) and stomatal index (SI; B, n = 254; D, n = 239) were measured in plants growing at their natural sites (nat. site) and in those cultivated in growth chambers under altitude-specific pCO₂ conditions. High altitude taxa (Anthyllis vulneraria subsp. valesiaca and Arabis alpina) grew at 2,970 m a.s.l. (pCO₂ approx. 29 Pa) and were cultivated at 30 Pa pCO₂, while the low altitude taxa (Anthyllis vulneraria subsp. carpatica and Arabidopsis thaliana) grew at 540 m a.s.l. (pCO₂ approx. 39 Pa) and were cultivated at 42 Pa pCO₂. Note: A. thaliana cultivated in the growth chambers at 42 Pa is the Col-0 wild-type. Marginal means along with their 95% confidence intervals are presented. The results of the statistical analysis are listed in S5 Table. Different letters denote statistically significant differences at p < 0.05, as determined by post hoc tests.

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Fig 5.

Stomatal frequency (SF) response to variations in pCO₂.

Stomatal density (SD; A, n = 836) and stomatal index (SI; B, n = 828) were determined for high-altitude taxa (“high”; altitude of origin 2’970 m a.s.l.), i.e., Anthyllis vulneraria subsp. valesiaca and Arabis alpina, and for low-altitude taxa (“low”; 540 m a.s.l.), i.e., Anthyllis vulneraria subsp. carpatica and Arabidopsis thaliana (Col-0). All plants were cultivated under reduced (30 Pa) and ambient pCO₂ (42 Pa). Marginal means along with their 95% confidence intervals are presented. Statistical analysis results are provided in S6 Table. Different letters denote statistically significant differences at p < 0.05, as determined by post hoc tests.

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Fig 6.

Response of above-ground fresh weight to variations in pCO2.

Fresh weight of the above-ground plant parts (n = 194) was measured in high-altitude taxa (“high”; altitude of origin 2’970 m a.s.l.), i.e., Anthyllis vulneraria subsp. valesiaca and Arabis alpina, and for low-altitude taxa (“low”; 540 m a.s.l.), i.e., Anthyllis vulneraria subsp. carpatica and Arabidopsis thaliana (Col-0). All plants were cultivated under reduced (30 Pa) and ambient pCO₂ (42 Pa). Marginal means along with their 95% confidence intervals are presented. Statistical analysis results are provided in S8 Table. Different letters denote statistically significant differences at p < 0.05, as determined by post hoc tests.

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Table 2.

Marginal effects of environmental factors on stomatal density.

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Table 2 Expand

Fig 7.

Marginal effects of environmental factors on stomatal density.

The effects of (A) partial pressure of CO2 (pCO₂), (B) air temperature, (C) vapor pressure deficit (VPD), and (D) irradiance on stomatal density (SD; n = 1’926) were analyzed in Arabis alpina and Arabidopsis thaliana (Col-0). The data were obtained from nine experiments conducted under environmental conditions that varied within the following minimum and maximum ranges: pCO₂ (29.1–43.3 Pa), temperature (16.7–22.5°C), VPD (10.3–13.7 hPa), and irradiance (85 and 175 µmol m ⁻ ² s ⁻ ¹). Marginal means along with their 95% confidence intervals are presented. Statistical analysis results are provided in Table 2.

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Table 3.

Marginal effects of environmental factors on stomatal index.

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Table 3 Expand

Fig 8.

Marginal effects of environmental factors on stomatal index.

The effects of (A) partial pressure of CO2 (pCO₂), (B) air temperature, (C) vapor pressure deficit (VPD), and (D) irradiance on stomatal index (SI; n = 490) were analyzed in Arabis alpina and Arabidopsis thaliana (Col-0). The data were obtained from six experiments conducted under environmental conditions that varied within the following minimum and maximum ranges: pCO₂ (29.5–43.3 Pa), temperature (19.0–21.3°C), VPD (10.4–13.7 hPa), and irradiance (85 and 175 µmol m ⁻ ² s ⁻ ¹). Marginal means along with their 95% confidence intervals are presented. Statistical analysis results are provided in Table 3.

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