Harnessing nighttime transpiration dynamics for drought tolerance in grasses.

Non-negligible nighttime transpiration rates (TRN) have been identified in grasses such as wheat and barley. Evidence from the last 30 years indicate that in drought-prone environments with high evaporative demand, TRN could amount to 8-55% of daytime TR, leading several investigators to hypothesize that reducing TRN might represent a viable water-saving strategy that minimizes seemingly 'wasteful' water loss that is not traded for CO2 fixation. More recently however, evidence suggests that actual increases in TRN during pre-dawn hours, which are presumably controlled by the circadian clock, mediate drought tolerance - not through water conservation - but by enabling maximized gas exchange early in the morning before midday depression sets in. Finally, new findings point to a previously undocumented role for leaf sheaths as substantial contributors (up to 45%) of canopy TRN, although the extent of their involvement in these two strategies remains unknown. In this paper, we synthesize and reconcile key results from experimental and simulation-based modeling efforts conducted at scales ranging from the leaf tissue to the field plot on wheat and barley to show that both strategies could in fact concomitantly enable yield gains under limited water supply. We propose a simple framework highlighting the role played by TRN dynamics in drought tolerance and provide a synthesis of potential research directions, with an emphasis on the need for further examining the role played by the circadian clock and leaf sheath gas exchange.

Sleep tight and wake-up early: nocturnal transpiration traits to increase wheat drought tolerance in a Mediterranean environment.

In wheat, night-time transpiration rate (TRN) could amount to 14-55% of daytime transpiration rate (TR), depending on the cultivar and environment. Recent evidence suggests that TRN is much less responsive to soil drying than daytime TR, and that such 'wasteful' water losses would increase the impact of drought on yields. In contrast, other evidence indicates that pre-dawn, circadian increases in TRN may enable enhanced radiation use efficiency, resulting in increased productivity under water deficit. Until now, there have been no attempts to evaluate these seemingly conflicting hypotheses in terms of their impact on yields in any crop. Here, using the Mediterranean environment of Tunisia as a case study, we undertook a simulation modelling approach using SSM-Wheat to evaluate yield outcomes resulting from these TRN trait modifications. TRN represented 15% of daytime TR-generated yield penalties of up to 20%, and these worsened when TRN was not sensitive to soil drying TR. For the same TRN level (15%), simulating a predawn increase in TRN alleviated yield penalties, leading to yield gains of up to 25%. Overall, this work suggests that decreasing TRN but increasing pre-dawn circadian control would be a viable breeding target to increase drought tolerance in a Mediterranean environment.

Variability in temperature-independent transpiration responses to evaporative demand correlate with nighttime water use and its circadian control across diverse wheat populations.

MAIN CONCLUSION: Nocturnal transpiration, through its circadian control, plays a role in modulating daytime transpiration response to increasing evaporative demand, to potentially enable drought tolerance in wheat. Limiting plant transpiration rate (TR) in response to increasing vapor pressure deficit (VPD) has been suggested to enable drought tolerance through water conservation. However, there is very little information on the extent of diversity of TR response curves to "true" VPD (i.e., independent from temperature). Furthermore, new evidence indicate that water-saving could operate by modulating nocturnal TR (TRN), and that this response might be coupled to daytime gas exchange. Based on 3 years of experimental data on a diverse group of 77 genotypes from 25 countries and 5 continents, a first goal of this study was to characterize the functional diversity in daytime TR responses to VPD and TRN in wheat. A second objective was to test the hypothesis that these traits could be coupled through the circadian clock. Using a new gravimetric phenotyping platform that allowed for independent temperature and VPD control, we identified three and fourfold variation in daytime and nighttime responses, respectively. In addition, TRN was found to be positively correlated with slopes of daytime TR responses to VPD, and we identified pre-dawn variation in TRN that likely mediated this relationship. Furthermore, pre-dawn increase in TRN positively correlated with the year of release among drought-tolerant Australian cultivars and with the VPD threshold at which they initiated water-saving. Overall, the study indicates a substantial diversity in TR responses to VPD that could be leveraged to enhance fitness under water-limited environments, and that TRN and its circadian control may play an important role in the expression of water-saving.

Potential involvement of root auxins in drought tolerance by modulating nocturnal and daytime water use in wheat.

BACKGROUND AND AIMS: The ability of wheat genotypes to save water by reducing their transpiration rate (TR) at times of the day with high vapour pressure deficit (VPD) has been linked to increasing yields in terminal drought environments. Further, recent evidence shows that reducing nocturnal transpiration (TRN) could amplify water saving. Previous research indicates that such traits involve a root-based hydraulic limitation, but the contribution of hormones, particularly auxin and abscisic acid (ABA), has not been explored to explain the shoot-root link. In this investigation, based on physiological, genetic and molecular evidence gathered on a mapping population, we hypothesized that root auxin accumulation regulates whole-plant water use during both times of the day. METHODS: Eight double-haploid lines were selected from a mapping population descending from two parents with contrasting water-saving strategies and root hydraulic properties. These spanned the entire range of slopes of TR responses to VPD and TRN encountered in the population. We examined daytime/night-time auxin and ABA contents in the roots and the leaves in relation to hydraulic traits that included whole-plant TR, plant hydraulic conductance (KPlant), slopes of TR responses to VPD and leaf-level anatomical traits. KEY RESULTS: Root auxin levels were consistently genotype-dependent in this group irrespective of experiments and times of the day. Daytime root auxin concentrations were found to be strongly and negatively correlated with daytime TR, KPlant and the slope of TR response to VPD. Night-time root auxin levels significantly and negatively correlated with TRN. In addition, daytime and night-time leaf auxin and ABA concentrations did not correlate with any of the examined traits. CONCLUSIONS: The above results indicate that accumulation of auxin in the root system reduces daytime and night-time water use and modulates plant hydraulic properties to enable the expression of water-saving traits that have been associated with enhanced yields under drought.

Pot binding as a variable confounding plant phenotype: theoretical derivation and experimental observations.

MAIN CONCLUSION: Theoretical derivation predicted growth retardation due to pot water limitations, i.e., pot binding. Experimental observations were consistent with these limitations. Combined, these results indicate a need for caution in high-throughput screening and phenotyping. Pot experiments are a mainstay in many plant studies, including the current emphasis on developing high-throughput, phenotyping systems. Pot studies can be vulnerable to decreased physiological activity of the plants particularly when pot volume is small, i.e., "pot binding". It is necessary to understand the conditions under which pot binding may exist to avoid the confounding influence of pot binding in interpreting experimental results. In this paper, a derivation is offered that gives well-defined conditions for the occurrence of pot binding based on restricted water availability. These results showed that not only are pot volume and plant size important variables, but the potting media is critical. Artificial potting mixtures used in many studies, including many high-throughput phenotyping systems, are particularly susceptible to the confounding influences of pot binding. Experimental studies for several crop species are presented that clearly show the existence of thresholds of plant leaf area at which various pot sizes and potting media result in the induction of pot binding even though there may be no immediate, visual plant symptoms. The derivation and experimental results showed that pot binding can readily occur in plant experiments if care is not given to have sufficiently large pots, suitable potting media, and maintenance of pot water status. Clear guidelines are provided for avoiding the confounding effects of water-limited pot binding in studying plant phenotype.

Nighttime evaporative demand induces plasticity in leaf and root hydraulic traits.

Increasing evidence suggests that nocturnal transpiration rate (TRN ) is a non-negligible contributor to global water cycles. Short-term variation in nocturnal vapor pressure deficit (VPDN ) has been suggested to be a key environmental variable influencing TRN . However, the long-term effects of VPDN on plant growth and development remain unknown, despite recent evidence documenting long-term effects of daytime VPD on plant anatomy, growth and productivity. Here we hypothesized that plant anatomical and functional traits influencing leaf and root hydraulics could be influenced by long-term exposure to VPDN . A total of 23 leaf and root traits were examined on four wheat (Triticum aestivum) genotypes, which were subjected to two long-term (30 day long) growth experiments where daytime VPD and daytime/nighttime temperature regimes were kept identical, with variation only stemming from VPDN , imposed at two levels (0.4 and 1.4 kPa). The VPDN treatment did not influence phenology, leaf areas, dry weights, number of tillers or their dry weights, consistently with a drought and temperature-independent treatment. In contrast, vein densities, adaxial stomata densities, TRN and cuticular TR, were strongly increased following exposure to high VPDN . Simultaneously, whole-root system xylem sap exudation and seminal root endodermis thickness were decreased, hypothetically indicating a change in root hydraulic properties. Overall these results suggest that plants 'sense' and adapt to variations in VPDN conditions over developmental scales by optimizing both leaf and root hydraulics.

High resolution mapping of traits related to whole-plant transpiration under increasing evaporative demand in wheat.

Atmospheric vapor pressure deficit (VPD) is a key component of drought and has a strong influence on yields. Whole-plant transpiration rate (TR) response to increasing VPD has been linked to drought tolerance in wheat, but because of its challenging phenotyping, its genetic basis remains unexplored. Further, the genetic control of other key traits linked to daytime TR such as leaf area, stomata densities and - more recently - nocturnal transpiration remains unknown. Considering the presence of wheat phenology genes that can interfere with drought tolerance, the aim of this investigation was to identify at an enhanced resolution the genetic basis of the above traits while investigating the effects of phenology genes Ppd-D1 and Ppd-B1 Virtually all traits were highly heritable (heritabilities from 0.61 to 0.91) and a total of mostly trait-specific 68 QTL were detected. Six QTL were identified for TR response to VPD, with one QTL (QSLP.ucl-5A) individually explaining 25.4% of the genetic variance. This QTL harbored several genes previously reported to be involved in ABA signaling, interaction with DREB2A and root hydraulics. Surprisingly, nocturnal TR and stomata densities on both leaf sides were characterized by highly specific and robust QTL. In addition, negative correlations were found between TR and leaf area suggesting trade-offs between these traits. Further, Ppd-D1 had strong but opposite effects on these traits, suggesting an involvement in this trade-off. Overall, these findings revealed novel genetic resources while suggesting a more direct role of phenology genes in enhancing wheat drought tolerance.

Genotype-dependent influence of night-time vapour pressure deficit on night-time transpiration and daytime gas exchange in wheat.

In crop plants, accumulating evidence indicates non-marginal night-time transpiration (TRNight) that is responsive to environmental conditions, especially in semiarid areas. However, the agronomical advantages resulting from such phenomenon remain obscure. Recently, drought-tolerance strategies directly stemming from daytime TR (TRDay) responses to daytime vapour pressure deficit VPD (VPDDay) were identified in wheat (Triticum spp.), but the existence of similar strategies resulting from TRNight response to night-time VPD (VPDNight) remains to be investigated, especially that preliminary evidence on this species indicates that TRNight might be responsive to VPDNight. Our study aims at investigating such strategies among a group of diverse lines including drought-tolerant genotypes. The study revealed that: (i) TRNight can be as high as 55% that of the maximal TRDay; (ii) VPDNight is the major driver of TRNight in a genotype-dependent fashion and has an impact on following daytime gas exchange; and (iii) a strong correlation exists between TR sensitivities to VPD under night-time and daytime conditions, revealing that tolerance strategies such as conservative water use do also exist under night-time environments. Overall, this report opens the way to further phenotyping and modelling work aiming at assessing the potential of using TRNight as a trait in breeding new drought-tolerant germplasm.