Focus on Water: Systems Analysis of Guard Cell Membrane Transport for Enhanced Stomatal Dynamics and Water Use Efficiency

Stomatal transpiration is at the center of a crisis in water availability and crop production that is expected to unfold over the next 20 to 30 years. Global water usage has increased 6-fold in the past 100 years, twice as fast as the human population, and is expected to double again before 2030, driven mainly by irrigation and agriculture. Guard cell membrane transport is integral to controlling stomatal aperture and offers important targets for genetic manipulation to improve crop performance. However, its complexity presents a formidable barrier to exploring such possibilities. With few exceptions, mutations that increase water use efficiency commonly have been found to do so with substantial costs to the rate of carbon assimilation, reflecting the trade-off in CO₂ availability with suppressed stomatal transpiration. One approach yet to be explored in detail relies on quantitative systems analysis of the guard cell. Our deep knowledge of transport and homeostasis in these cells gives real substance to the prospect for reverse engineering of stomatal responses, using in silico design in directing genetic manipulation for improved water use and crop yields. Here we address this problem with a focus on stomatal kinetics, taking advantage of the OnGuard software and models of the stomatal guard cell recently developed for exploring stomatal physiology. Our analysis suggests that manipulations of single transporter populations are likely to have unforeseen consequences. Channel gating, especially of the dominant K⁺ channels, appears the most favorable target for experimental manipulation.Stomata are pores that provide the major route for gaseous exchange across the impermeable cuticle of leaves and stems (Hetherington and Woodward, 2003). They open and close in response to exogenous and endogenous signals and thereby control the exchange of gases, most importantly water vapor and CO₂, between the interior of the leaf and the atmosphere. Stomata exert major controls on the water and carbon cycles of the world (Schimel et al., 2001) and can limit photosynthetic rates by 50% or more when demand exceeds water supply (Ni and Pallardy, 1992). Stomatal transpiration is at the center of a crisis in water availability and crop production that is expected to unfold over the next 20 to 30 years; indeed, global water usage has increased 6-fold in the past 100 years, twice as fast as the human population, and is expected to double again before 2030, driven mainly by irrigation and agriculture (United Nations Educational, Scientific and Cultural Organization, 2009).Guard cell transport is integral to controlling stomatal aperture. Guard cells surround the stomatal pore and respond in a well-defined manner to an array of extracellular signals, including light, to regulate its aperture. Guard cells coordinate membrane transport within a complex network of intracellular signals (Willmer and Fricker, 1996; Blatt, 2000a, 2000b; Hetherington and Woodward, 2003; Shimazaki et al., 2007) to regulate fluxes, mainly of K⁺, Cl⁻, and malate, driving cell turgor and stomatal aperture. Our deep knowledge of these processes has made the guard cell the best known of plant cell models for membrane transport, signaling, and homeostasis (Willmer and Fricker, 1996; Blatt, 2000b; Roelfsema and Hedrich, 2010; Hills et al., 2012). This knowledge gives real substance to the prospect for reverse engineering of stomatal responses, using in silico design in directing genetic manipulation for improved crop yields, especially under water-limited conditions.Water use efficiency (WUE; defined as the amount of dry matter produced per unit of water transpired) is directly related to stomatal function. Thus, at the practical level, stomata represent an important target for breeders interested in manipulating crop performance. A large body of data relates stomata, transpiration, and carbon assimilation (Willmer and Fricker, 1996; Farquhar et al., 2001; Hetherington and Woodward, 2003; Lawson et al., 2011). Several examples illustrate how manipulating of stomatal characteristics can affect WUE (Fischer et al., 1998; Rebetzke et al., 2002; Masle et al., 2005; Eisenach et al., 2012). With few exceptions, however, mutations that increase WUE commonly do so at the expense of carbon assimilation, reflecting the trade-off in CO₂ availability with suppressed stomatal transpiration.Stomatal movements generally lag behind short-term changes in available light associated with sunflecks and shadeflecks (Pearcy, 1990; Lawson et al., 2012; Lawson and Blatt, 2014). This hysteresis in response, between stomatal aperture and gas exchange on one hand and photosynthetic capacity on the other, can lead alternately to periods of assimilation limited by stomatal conductance, and of high transpiration without corresponding rates of assimilation (Lawson et al., 2011). It has been argued that such hysteresis in stomatal responsiveness with the demand for CO₂ erodes assimilation and WUE, with substantial consequences for long-term yield (Vico et al., 2011; Eisenach et al., 2012; Lawson et al., 2012; Lawson and Blatt, 2014). If so, then improving WUE with gains in assimilation should be possible if the speed of stomatal responsiveness can be enhanced. However, the complexity of guard cell transport presents a formidable barrier to exploring such possibilities. Here we address this problem, taking advantage of OnGuard models of the stomatal guard cell. We explore in silico the potential for enhancing stomatal kinetics through single transporter (single gene product) manipulations. Our results identify the gating of the dominant K⁺ channels as the most promising target for experimental manipulation.