Biology subjects

Twohey, R. J.

Publications and source records attributed to Twohey, R. J..

5 recordsLinked to original sources

bioRxiv2025

A spontaneous mutation in a key C4 pathway gene significantly alters leaf δ13C uncoupling its relationship with WUE and photosynthetic performance in Zea mays

Increases in global temperature and drought are negatively impacting the yields of major crops. Therefore, targeted improvements to intrinsic water use efficiency (WUEi) are needed to reduce the water required for agricultural production. While it is very time-consuming to directly measure WUEi, stable carbon isotope ratios ({delta}13C) are a reliable high throughput proxy trait for quantifying WUEi in C3 species. While genetic studies have improved our understanding of the relationship between WUEi and {delta}13C in C4 species, the knowledge needed to implement {delta}13C in breeding schemes is incomplete. Using a Zea mays line with an extremely negative {delta}13C value, a quantitative genetics approach was used to identify a large deletion in carbonic anhydrase1 (cah1). Carbonic anhydrase is the first enzymatic step of the C4 photosynthetic pathway and is known to affect {delta}13C. Surprisingly, the line with the mutant allele has significantly higher carbonic anhydrase activity with a concurrent reduction in {delta}13C, opposite of what would be expected based on C4 carbon isotope fractionation theory. These observed decouple {delta}13C and WUEi, which calls for further investigation into carbon isotope discrimination in C4 species.

plant biology↗

bioRxiv2025

Loss of plasma membrane conductance in outside-xylem zone explains non-stomatal control of transpiration

The conventional assumption is that stomatal conductance (gs) dominates the regulation of water and carbon dioxide fluxes between leaves and the atmosphere. Here, a nanoreporter of water status at the mesophyll cell surface and local xylem within intact maize leaves documents significant undersaturation of water vapor in the outside-xylem zone (OXZ) and a large loss of conductance of this zone (goxz) at moderate xylem water stress (no turgor loss). The ratio of the resistances (1/goxz)/(1/gs) serves as a predictive phenotype of undersaturation, non-stomatal regulation of transpiration, errors in standard gas exchange analysis, and an increase of intrinsic water use efficiency (iWUE). Cell-scale access to water status reveals symplasmic-apoplasmic disequilibrium and informs a biophysical model that can explain experimental observations quantitatively based on localization of variable conductance to the plasma membrane. This work opens new paths of inquiry into the molecular basis and functional consequences of non-stomatal regulation of transpiration.

plant biology↗

bioRxiv2025

Investigating the role of stomatal dynamics on agronomic traits using a slac1-2 Zea mays mutant

The production of staple food crops is becoming increasingly difficult due to the amount of freshwater needed to realize food security for a growing global population. Climate projections show hotter and drier growing seasons in traditionally productive agricultural regions, which will increase the demand for limited water resources. Thus, strategies to improve water use efficiency will only become more important for sustainable agriculture. At the leaf level, stomatal conductance has a large influence on transpirational water loss and therefore water use efficiency. The anion channel SLAC1 has been shown in several plant species to function as the primary mechanism for stomatal closure, which allows stomatal aperture to change dynamically in response to environmental stimuli. Given that slac1 is a single gene that can significantly alter stomatal conductance, it has the potential to serve as a control point for improving water use efficiency. Here we fully characterize a Zea mays slac1-2 mutant in multiple field environments. Interestingly, homozygous slac1-2 hybrids did not show improved net CO2 assimilation or increased grain yield despite having greater stomatal conductance compared to a wild-type hybrid. Net CO2 assimilation and grain yield were either lower or similar in slac1-2 compared to wild-type across environments. Furthermore, the slac1-2 hybrid did not have increased nitrogen uptake. These results suggest that the C4 carbon concentrating mechanism removes any stomatal conductance limitations to CO2 assimilation, even in highly productive wild-type Z. mays hybrids. Because the slac1-2 hybrids eliminate stomatal conductance as a major variable for modulating water loss, future studies will be able to investigate alternate regulators of plant water potential to identify novel mechanisms for increasing water use efficiency.

physiology↗

bioRxiv2023

Differences in stomatal sensitivity to COC2 and light influences variation in water use efficiency and leaf carbon isotope composition in two genotypes of the C4 plant Zea mays.

The ratio of net CO2 uptake (Anet) and stomatal conductance (gs) is an intrinsic measurement of leaf water use efficiency (WUEi) however its measurement can be challenging for large phenotypic screens. Measurements of leaf carbon isotope composition ({delta}13Cleaf) may be a scalable tool to approximate WUEi for screening because it in part reflects the competing influences of Anet and gs on the CO2 partial pressure (pCO2) inside the leaf over time. However, in C4 photosynthesis the CO2 concentrating mechanism complicates the relationship between {delta}13Cleaf and WUEi. Despite this complicated relationship, several studies have shown genetic variation in {delta}13Cleaf across C4 plants. Yet there has not been a clear demonstration of whether Anet or gs are the causal mechanisms controlling WUEi and {delta}13Cleaf. Our approach was to characterize leaf photosynthetic traits of two Zea mays recombinant inbred lines (Z007E0067 and Z007E0150) which consistently differ for {delta}13Cleaf even though they have minimal confounding genetic differences. We demonstrate that these two genotypes contrasted in WUEi driven by differences in the speed of stomatal responses to changes in pCO2 and light that lead to unproductive leaf water loss. These findings provide support that differences in {delta}13Cleaf in closely related genotypes do reflect greater WUEi and further suggests that differences in stomatal kinetic response to changing environmental conditions is a key target to improve WUEi.

plant biology↗

bioRxiv2018

Stable Carbon Isotopes in Zea mays

The increasing demand for food production and predicted climate change scenarios highlight the need for improvements in crop sustainability. The efficient use of water will become increasingly important for rainfed agricultural crops even in fertile regions that have historically received ample precipitation. Improvements in water-use efficiency in Zea mays have been limited, and warrants a renewed effort aided by molecular breeding approaches. Progress has been constrained by the difficulty of measuring water-use in a field environment. The stable carbon isotope composition ({delta}13C) of the leaf has been proposed as an integrated signature of carbon fixation with a link to stomatal conductance. However, additional factors affecting leaf {delta}13C exist, and a limited number of studies have explored this trait in Z. mays. Here we present an extensive characterization of leaf {delta}13C in Z. mays. Significant variation in leaf {delta}13C exists across diverse lines of Z. mays, which we show to be heritable across several environments.\n\nFurthermore, we examine temporal and spatial variation in leaf {delta}13C to determine the optimum sampling time to maximize the use of leaf {delta}13C as a trait. Finally, our results demonstrate the relationship between transpiration and leaf {delta}13C in the field and the greenhouse. Decreasing transpiration and soil moisture are associated with decreasing leaf {delta}13C. Taken together these results outline a strategy for using leaf {delta}13C and reveal its usefulness as a measure of transpiration efficiency under well-watered conditions rather than a predictor of performance under drought.\n\nSignificance StatementThis study identifies sources of variation in stable carbon isotopes of maize leaves and establishes the framework for connecting leaf {delta}13C and transpiration efficiency.

plant biology↗