Caroline Elliott-Kingston

Stomatal control as a driver of plant evolution

Stomata are the pores on a leaf surface through which plants regulate the uptake of carbon dioxide (CO2) for photosynthesis against the loss of water via transpiration. Turgor changes in the guard cells determine the area of stomatal pore through which gaseous diffusion can occur, thus maintaining a constant internal environment within the leaf (Gregory et al., 1950). Stomata first occurred in the fossil record ;400 million years ago (Ma), and are largely identical in form to the stomatal complexes of many extant plants, illustrating their effectiveness and importance to terrestrial plants (Edwards et al., 1998). Stomatal control is critical to a plant’s adaptation to its environment; it is this fundamental importance that has led to a wealth of stomatal research ranging in scale from biomolecular analysis to landscape processes (e.g. Gedney et al., 2006; Hu et al., 2010). The first issue of Journal of Experimental Botany, published 60 years ago, contained four papers relating to stomatal function. These included an analysis by Heath of the effects of atmospheric CO2 concentration ([CO2]) on stomatal aperture and conductance; an area of research that is increasingly relevant to our understanding of the past and prediction of future vegetation responses to atmospheric composition. Heath (1950) was the first to observe that reductions in [CO2] below ambient levels induced stomatal opening, an ecophysiological response of great interest, and that the site of CO2 sensing was most probably in the substomatal cavity and not the guard cells. Stomatal research has become vastly important to crop production, biodiversity responses, and hydrology (particularly in terms of ‘run-off’) with respect to rising atmospheric [CO2], changing water regimes, and growing populations. As our understanding of stomatal physiology develops, the role of stomata in the evolution of terrestrial vegetation and development of the terrestrial landscape and atmospheric composition is becoming increasingly evident, alongside the use of fossil stomata as palaeo-proxies of past atmospheres (e.g. McElwain et al., 2004; Berry et al., 2010; Smith et al., 2010). The stomatal control responses of plants consist of ‘shortterm’ stomatal aperture changes in response to availability of water, light, temperature, wind speed, and carbon dioxide, and also ‘longer term’ changes in stomatal density that set the limits for maximum stomatal conductance in response to atmospheric [CO2], light intensity/quality, and root-to-shoot signals of water availability (Schoch et al., 1980, 1984; Davies et al., 2000; Casson et al., 2009). Stomatal control determines the water use efficiency (WUE) of a plant by optimizing water lost against carbon gained. Additionally, the stomatal control mechanisms employed by a plant species will determine: the risk of xylem embolism by reducing the probability of cavitation through stomatal closure during episodes of high transpirative demand (Brodribb and Jordan, 2008; Meinzer et al., 2009); leaf temperature and resistance to heat stress (Srivastava et al., 1995; Jones et al., 2002); tolerance of toxic atmospheric gases (Mansfield and Majernik, 1970); nutrient uptake via promotion of root mass flow (Van Vuuren et al., 1997); and the maximum rate of photosynthesis (Korner et al., 1979). Those plant species with more effective stomatal control will be expected to be more successful than those with less effective stomatal control. However, not all plant species, or individuals within a species, possess equally effective stomatal control, in the setting of either stomatal numbers or the regulation of stomatal aperture (i.e. speed and ‘tightness’ of closure). Given that any trait that confers a selective advantage is likely to become universal within a population (McNeilly, 1968), it may be reasonable to assume that stomatal control incurs certain ‘costs’, and that these costs have played a significant role in plant evolution over the last 400 million years. The origination of major plant groups, and morphological advances such as the development of planate leaves, coincide with periods of ‘low’ atmospheric [CO2] (Fig. 1) (Woodward, 1998; Beerling et al., 2001). The reduced availability of the substrate for photosynthesis is predicted to be compensated by increases in the carboxylation efficiency of RubisCO and enhanced stomatal conductance to maintain CO2 uptake during periods of low [CO2] (Woodward, 1998; Franks and Beerling, 2009). This elevated stomatal conductance incurs higher rates of water loss and associated risks of desiccation and xylem embolism, in addition to the metabolic costs of enhanced construction of stomatal complexes. It is these costs during periods of low [CO2] that may serve as evolutionary tipping points, where species with more efficient and effective stomata and hydraulic systems are favoured (Robinson, 1994; Brodribb

Does size matter? Atmospheric CO2 may be a stronger driver of stomatal closing rate than stomatal size in taxa that diversified under low CO2

One strategy for plants to optimize stomatal function is to open and close their stomata quickly in response to environmental signals. It is generally assumed that small stomata can alter aperture faster than large stomata. We tested the hypothesis that species with small stomata close faster than species with larger stomata in response to darkness by comparing rate of stomatal closure across an evolutionary range of species including ferns, cycads, conifers, and angiosperms under controlled ambient conditions (380 ppm CO2; 20.9% O2). The two species with fastest half-closure time and the two species with slowest half-closure time had large stomata while the remaining three species had small stomata, implying that closing rate was not correlated with stomatal size in these species. Neither was response time correlated with stomatal density, phylogeny, functional group, or life strategy. Our results suggest that past atmospheric CO2 concentration during time of taxa diversification may influence stomatal response time. We show that species which last diversified under low or declining atmospheric CO2 concentration close stomata faster than species that last diversified in a high CO2 world. Low atmospheric [CO2] during taxa diversification may have placed a selection pressure on plants to accelerate stomatal closing to maintain adequate internal CO2 and optimize water use efficiency.

How well do you know your growth chambers? Testing for chamber effect using plant traits

BackgroundPlant growth chambers provide a controlled environment to analyse the effects of environmental parameters (light, temperature, atmospheric gas composition etc.) on plant function. However, it has been shown that a ‘chamber effect’ may exist whereby results observed are not due to an experimental treatment but to inconspicuous differences in supposedly identical chambers. In this study, Vicia faba L. ‘Aquadulce Claudia’ (broad bean) plants were grown in eight walk-in chambers to establish if a chamber effect existed, and if so, what plant traits are best for detecting such an effect. A range of techniques were used to measure differences between chamber plants, including chlorophyll fluorescence measurements, gas exchange analysis, biomass, reproductive yield, anatomical traits and leaf stable carbon isotopes.Results and discussionFour of the eight chambers exhibited a chamber effect. In particular, we identified two types of chamber effect which we term ‘resolvable’ or ‘unresolved’; a resolvable chamber effect is caused by malfunctioning components of a chamber and an unresolved chamber effect is caused by unknown factors that can only be mitigated by appropriate experimental design and sufficient replication. Not all measured plant traits were able to detect a chamber effect and no single trait was capable of detecting all chamber effects. Fresh weight and flower count detected a chamber effect in three chambers, stable carbon isotopes (δ13C) and net rate CO2 assimilation (An) identified a chamber effect in two chambers, stomatal conductance (gs) and total performance index detected an effect only in one chamber.Conclusion(1) Chamber effects can be adequately detected by fresh weight measurements and flower counts on Vicia faba plants. These methods were the most effective in terms of detection and most efficient in terms of time. (2) δ13C, gs and An measurements help distinguish between resolvable and unresolved chamber effects. (3) Unresolved chamber effects require experimental unit replication while resolvable chamber effects require investigation, repair and retesting in advance of initiating further experiments.

Cuticle surfaces of fossil plants as a potential proxy for volcanic SO2 emissions: observations from the Triassic–Jurassic transition of East Greenland

Flood basalt volcanism has been implicated in several episodes of mass extinctions and environmental degradation in the geological past, including at the Triassic–Jurassic (Tr–J) transition, through global warming caused by massive outgassing of carbon dioxide. However, the patterns of biodiversity loss observed are complicated and sometimes difficult to reconcile with the effects of global warming alone. Recently, attention has turned to additional volcanic products as potential aggravating factors, in particular sulphur dioxide (SO2). SO2 acts both directly as a noxious environmental pollutant and indirectly through forming aerosols in the atmosphere, which may cause transient global dimming and cooling. Here, we present a range of morphological changes to fossil plant leaf cuticle surfaces of hundreds of Ginkgoales and Bennettitales specimens across the Tr–J boundary of East Greenland. Our results indicate that morphological structures of distorted cuticles near the Tr–J boundary are consistent with modern cuticle SO2-caused damage and supported by recent leaf-shape SO2 proxy results, thus identifying cuticle surface morphology as a potentially powerful proxy for SO2. Recording the timing and duration of SO2 emissions in the past may help distinguish between the driving agents responsible for mass extinction events and thus improve our understanding of the Earth System.

Co-ordination in Morphological Leaf Traits of Early Diverging Angiosperms Is Maintained Following Exposure to Experimental Palaeo-atmospheric Conditions of Sub-ambient O2 and Elevated CO2

In order to be successful in a given environment a plant should invest in a vein network and stomatal distribution that ensures balance between both water supply and demand. Vein density (Dv) and stomatal density (SD) have been shown to be strongly positively correlated in response to a range of environmental variables in more recently evolved plant species, but the extent of this relationship has not been confirmed in earlier diverging plant lineages. In order to examine the effect of a changing atmosphere on the relationship between Dv and SD, five early-diverging plant species representing two different reproductive plant grades were grown for 7 months in a palaeo-treatment comprising an O2:CO2 ratio that has occurred multiple times throughout plant evolutionary history. Results show a range of species-specific Dv and SD responses to the palaeo-treatment, however, we show that the strong relationship between Dv and SD under modern ambient atmospheric composition is maintained following exposure to the palaeo-treatment. This suggests strong inter-specific co-ordination between vein and stomatal traits for our study species even under relatively extreme environmental change. This co-ordination supports existing plant function proxies that use the distance between vein endings and stomata (Dm) to infer plant palaeo-physiology.