Matthew E. Gilbert

The role of bundle sheath extensions and life form in stomatal responses to leaf water status

Bundle sheath extensions (BSEs) are key features of leaf structure with currently little-understood functions. To test the hypothesis that BSEs reduce the hydraulic resistance from the bundle sheath to the epidermis (rbe) and thereby accelerate hydropassive stomatal movements, we compared stomatal responses with reduced humidity and leaf excision among 20 species with heterobaric or homobaric leaves and herbaceous or woody life forms. We hypothesized that low rbe due to the presence of BSEs would increase the rate of stomatal opening (V) during transient wrong-way responses, but more so during wrong-way responses to excision (Ve) than humidity (Vh), thus increasing the ratio of Ve to Vh. We predicted the same trends for herbaceous relative to woody species given greater hydraulic resistance in woody species. We found that Ve, Vh, and their ratio were 2.3 to 4.4 times greater in heterobaric than homobaric leaves and 2.0 to 3.1 times greater in herbaceous than woody species. To assess possible causes for these differences, we simulated these experiments in a dynamic compartment/resistance model, which predicted larger Ve and Ve/Vh in leaves with smaller rbe. These results support the hypothesis that BSEs reduce rbe. Comparison of our data and simulations suggested that rbe is approximately 4 to 16 times larger in homobaric than heterobaric leaves. Our study provides new evidence that variations in the distribution of hydraulic resistance within the leaf and plant are central to understanding dynamic stomatal responses to water status and their ecological correlates and that BSEs play several key roles in the functional ecology of heterobaric leaves.

Drought Adaptation Mechanisms Should Guide Experimental Design

The mechanism, or hypothesis, of how a plant might be adapted to drought should strongly influence experimental design. For instance, an experiment testing for water conservation should be distinct from a damage-tolerance evaluation. We define here four new, general mechanisms for plant adaptation to drought such that experiments can be more easily designed based upon the definitions. A series of experimental methods are suggested together with appropriate physiological measurements related to the drought adaptation mechanisms. The suggestion is made that the experimental manipulation should match the rate, length, and severity of soil water deficit (SWD) necessary to test the hypothesized type of drought adaptation mechanism.

Physiological trade-offs of stomatal closure under high evaporative gradients in field grown soybean

Limited rainfall is the main constraint to agriculture, making agricultural research to understand plant behaviour that leads to avoidance of soil water deficit a matter of priority. One focus has screened for crop varieties that decrease stomatal conductance under high vapour pressure deficit (VPD), a proxy for the leaf evaporative gradient. However, the link between stomatal closure and physiological consequences in field environments is not yet clear. A field experiment on soybeans demonstrated that considerable variation in leaf temperature relative to air temperature occurred, leading to evaporative gradients differing substantially from VPD. Thus, transpiration is decreased by stomatal closure at high VPD, but to compensate, transpiration is somewhat increased due to higher leaf temperatures. Soil water deficit led to lower stomatal conductance, particularly under low evaporative conditions, not just under hot conditions. Non-stomatal photosynthetic limitations were observed due to combined occurrence of stomatal closure and high temperature under high VPD. Although leaves reached temperatures higher than the threshold for a decrease in maximum photochemical efficiency, and displayed non-stomatal photosynthetic limitations, no photoinhibition or damage was observed by night-time. The results demonstrate that more understanding of physiological strategies for achieving altered water use is needed to avoid trade-offs and heat stress.

The bias of a two‐dimensional view: comparing two‐dimensional and three‐dimensional mesophyll surface area estimates using noninvasive imaging

The mesophyll surface area exposed to intercellular air space per leaf area (Sm ) is closely associated with CO2 diffusion and photosynthetic rates. Sm is typically estimated from two-dimensional (2D) leaf sections and corrected for the three-dimensional (3D) geometry of mesophyll cells, leading to potential differences between the estimated and actual cell surface area. Here, we examined how 2D methods used for estimating Sm compare with 3D values obtained from high-resolution X-ray microcomputed tomography (microCT) for 23 plant species, with broad phylogenetic and anatomical coverage. Relative to 3D, uncorrected 2D Sm estimates were, on average, 15-30% lower. Two of the four 2D Sm methods typically fell within 10% of 3D values. For most species, only a few 2D slices were needed to accurately estimate Sm within 10% of the whole leaf sample median. However, leaves with reticulate vein networks required more sections because of a more heterogeneous vein coverage across slices. These results provide the first comparison of the accuracy of 2D methods in estimating the complex 3D geometry of internal leaf surfaces. Because microCT is not readily available, we provide guidance for using standard light microscopy techniques, as well as recommending standardization of reporting Sm values.

Excess diffuse light absorption in upper mesophyll limits CO2 drawdown and depresses photosynthesis

Excess absorption of diffuse versus direct light in the upper mesophyll of Helianthus annuus sun leaves leads to suboptimal photosynthesis. In agricultural and natural systems, diffuse light can enhance plant primary productivity due to deeper penetration into and greater irradiance of the entire canopy. However, for individual sun-grown leaves from three species, photosynthesis is actually less efficient under diffuse compared with direct light. Despite its potential impact on canopy-level productivity, the mechanism for this leaf-level diffuse light photosynthetic depression effect is unknown. Here, we investigate if the spatial distribution of light absorption relative to electron transport capacity in sun- and shade-grown sunflower (Helianthus annuus) leaves underlies its previously observed diffuse light photosynthetic depression. Using a new one-dimensional porous medium finite element gas-exchange model parameterized with light absorption profiles, we found that weaker penetration of diffuse versus direct light into the mesophyll of sun-grown sunflower leaves led to a more heterogenous saturation of electron transport capacity and lowered its CO2 concentration drawdown capacity in the intercellular airspace and chloroplast stroma. This decoupling of light availability from photosynthetic capacity under diffuse light is sufficient to generate an 11% decline in photosynthesis in sun-grown but not shade-grown leaves, primarily because thin shade-grown leaves similarly distribute diffuse and direct light throughout the mesophyll. Finally, we illustrate how diffuse light photosynthetic depression could overcome enhancement in canopies with low light extinction coefficients and/or leaf area, pointing toward a novel direction for future research.

Beyond porosity: 3D leaf intercellular airspace traits that impact mesophyll conductance

The gas phase of mesophyll conductance is affected by the 3D traits tortuosity, path lengthening, and airspace connectivity, in addition to porosity. The leaf intercellular airspace (IAS) is generally considered to have high conductance to CO2 diffusion relative to the liquid phase. While previous studies accounted for leaf-level variation in porosity and mesophyll thickness, they omitted 3D IAS traits that potentially influence IAS conductance (gIAS). Here, we reevaluated the standard equation for gIAS by incorporating tortuosity, lateral path lengthening, and IAS connectivity. We measured and spatially mapped these geometric IAS traits for 19 Bromeliaceae species with Crassulacean acid metabolism (CAM) or C3 photosynthetic pathways using x-ray microcomputed tomography imaging and a novel computational approach. We found substantial variation in porosity (0.04–0.73 m3 m−3), tortuosity (1.09–3.33 m2 m−2), lateral path lengthening (1.12–3.19 m m−1), and IAS connectivity (0.81–0.97 m2 m−2) across all bromeliad leaves. The revised gIAS model predicted significantly lower gIAS in CAM (0.01–0.19 mol m−2 s−1 bar−1) than in C3 (0.41–2.38 mol m−2 s−1 bar−1) plants due to a coordinated decline in these IAS traits. Our reevaluated equation also generally predicted lower gIAS values than the former one. Moreover, we observed high spatial heterogeneity in these IAS geometric traits throughout the mesophyll, especially within CAM leaves. Our data show that IAS traits that better capture the 3D complexity of leaves strongly influence gIAS and that the impact of the IAS on mesophyll conductance should be carefully considered with respect to leaf anatomy. We provide a simple function to estimate tortuosity and lateral path lengthening in the absence of access to imaging tools such as x-ray microcomputed tomography or other novel 3D image-processing techniques.