François Tardieu

Combining Quantitative Trait Loci Analysis and an Ecophysiological Model to Analyze the Genetic Variability of the Responses of Maize Leaf Growth to Temperature and Water Deficit

Ecophysiological models predict quantitative traits of one genotype in any environment, whereas quantitative trait locus (QTL) models predict the contribution of alleles to quantitative traits under a limited number of environments. We have combined both approaches by dissecting into effects of QTLs the parameters of a model of maize (Zea mays) leaf elongation rate (LER; H. Ben Haj Salah, F. Tardieu [1997] Plant Physiol 114: 893–900). Response curves of LER to meristem temperature, water vapor pressure difference, and soil water status were established in 100 recombinant inbred lines (RILs) of maize in six experiments carried out in the field or in the greenhouse. All responses were linear and common to different experiments, consistent with the model. A QTL analysis was carried out on the slopes of these responses by composite interval mapping confirmed by bootstrap analysis. Most QTLs were specific of one response only. QTLs of abscisic acid concentration in the xylem sap colocalized with QTLs of response to soil water deficit and conferred a low response. Each parameter of the ecophysiological model was computed as the sum of QTL effects, allowing calculation of parameters for 11 new RILs and two parental lines. LERs were simulated and compared with measurements in a growth chamber experiment. The combined model accounted for 74% of the variability of LER, suggesting that it has a general value for any RIL under any environment.

Drought and Abscisic Acid Effects on Aquaporin Content Translate into Changes in Hydraulic Conductivity and Leaf Growth Rate: A Trans-Scale Approach

The effects of abscisic acid (ABA) on aquaporin content, root hydraulic conductivity (Lpr), whole plant hydraulic conductance, and leaf growth are controversial. We addressed these effects via a combination of experiments at different scales of plant organization and tested their consistency via a model. We analyzed under moderate water deficit a series of transformed maize (Zea mays) lines, one sense and three antisense, affected in NCED (for 9-cis-epoxycarotenoid dioxygenase) gene expression and that differed in the concentration of ABA in the xylem sap. In roots, the mRNA expression of most aquaporin PIP (for plasma membrane intrinsic protein) genes was increased in sense plants and decreased in antisense plants. The same pattern was observed for the protein contents of four PIPs. This resulted in more than 6-fold differences between lines in Lpr under both hydrostatic and osmotic gradients of water potential. This effect was probably due to differences in aquaporin activity, because it was nearly abolished by a hydrogen peroxide treatment, which blocks the water channel activity of aquaporins. The hydraulic conductance of intact whole plants was affected in the same way when measured either in steady-state conditions or via the rate of recovery of leaf water potential after rewatering. The recoveries of leaf water potential and elongation upon rehydration differed between lines and were accounted for by the experimentally measured Lpr in a model of water transfer. Overall, these results suggest that ABA has long-lasting effects on plant hydraulic properties via aquaporin activity, which contributes to the maintenance of a favorable plant water status.

Analysis of the spatial variability of maize root density

The spatial arrangement of maize roots was studied in a clay loam field in order to test the regularity of root arrangement, which is implicitly assumed when distances between neighbouring roots are calculated. For that. we carried out a mapping of root contacts on six superposed horizontal planes which cut the rooting volume of several area samples. Three situations were studied: (i) one inter-row out of two was compacted down to the base of the ploughed layer (28 cm), but not in non-tilled layers (28 to 200 cm); (ii) a mechanical obstacle was placed at the base of the ploughed layer; (iii) one inter-row out of two was compacted down to half the depth of the ploughed layer. On all horizontal planes, the spatial arrangement or root contacts followed a non-regular, clustered pattern for a 10−2m scale of study, even in parts of soil which had not been disturbed by compactions. In the first two situations, where obstacles met the base of the ploughed layer, root density in non-tilled layers was several times lower below the obstacles than below the remaining parts of the ploughed layer. This caused a 10−1 m sized variability which was superimposed on to the 10−2 m one. Conversely in the third situation, obstacles had no appreciable effect on root density in non-tilled layers. Obstacles located at the base of the ploughed layer therefore prevented root access to non-tilled layer and caused a ‘shadow effect’ in the non-tilled layers. This effect is probably due to the main vertical direction of roots in these layers.

Control of leaf growth by abscisic acid: hydraulic or non‐hydraulic processes?

Abscisic acid (ABA) affects plant metabolism and water transfers via multiple mechanisms at cell, organ and whole plant levels. These mechanisms translate into contradictory effects on leaf growth, so the literature reports positive, null or negative effects of ABA on leaf growth upon water deficit. We review evidences based on genetic manipulations of ABA biosynthesis, feeding the plant with artificial ABA or partial root drying and provide elements to avoid confusions of effects. We propose that ABA has mainly three effects on growth. (i) Via its controlling effect on stomatal aperture and transpiration rate, an increased concentration of ABA tends to buffer the day-night alternations of leaf growth rate and the negative effect of evaporative demand. (ii) ABA tends to improve leaf growth via an increase in the conductance to water transfer in the plant as a result of increased tissue hydraulic conductivity. (iii) ABA has also a modest non-hydraulic effect which is negative in plants subjected to water deficit, either manipulated for ABA synthesis or fed with artificial ABA, but can be positive in well watered plants deficient of ABA. The overall effect of increasing ABA biosynthesis depends on the relative weight of each of these effects under different environmental scenarios.

Aquaporin-mediated reduction in maize root hydraulic conductivity impacts cell turgor and leaf elongation even without changing transpiration

Root hydraulic conductivity in plants (Lpr) exhibits large variations in response to abiotic stimuli. In this study, we investigated the impact of dynamic, aquaporin-mediated changes of Lpr on leaf growth, water potential, and water flux throughout the plant. For this, we manipulated Lpr by subjecting roots to four independent treatments, with aquaporin inhibitors applied either to transpiring maize (Zea mays) plants grown in hydroponics or to detopped root systems for estimation of Lpr. The treatments were acid load at pH 6.0 and 5.0 and hydrogen peroxide and anoxia applied for 1 to 2 h and subsequently reversed. First, we established that acid load affected cell hydraulic conductivity in maize root cortex. Lpr was reduced by all treatments by 31% to 63%, with half-times of about 15 min, and partly recovered when treatments were reversed. Cell turgor measured in the elongating zone of leaves decreased synchronously with Lpr, and leaf elongation rate closely followed these changes across all treatments in a dose-dependent manner. Leaf and xylem water potentials also followed changes in Lpr. Stomatal conductance and rates of transpiration and water uptake were not affected by Lpr reduction under low evaporative demand. Increased evaporative demand, when combined with acid load at pH 6.0, induced stomatal closure and amplified all other responses without altering their synchrony. Root pressurization reversed the impact of acid load or anoxia on leaf elongation rate and water potential, further indicating that changes in turgor mediated the response of leaf growth to reductions in Lpr.

Association of specific expansins with growth in maize leaves is maintained under environmental, genetic, and developmental sources of variation

We aimed to evaluate whether changes in maize (Zea mays) leaf expansion rate in response to environmental stimuli or developmental gradients are mediated by common or specific expansins, a class of proteins known to enhance cell wall extensibility. Among the 33 maize expansin or putative expansin genes analyzed, 19 were preferentially expressed at some point of the leaf elongation zone and these expansins could be organized into three clusters related to cell division, maximal leaf expansion, and cell wall differentiation. Further analysis of the spatial distribution of expression was carried out for three expansins in leaves displaying a large range of expansion rates due to water deficit, genotype, and leaf developmental stage. With most sources of variation, the three genes showed similar changes in expression and consistent association with changes in leaf expansion. Moreover, our analysis also suggested preferential association of each expansin with elongation, widening, or both of these processes. Finally, using in situ hybridization, expression of two of these genes was increased in load-bearing tissues such as the epidermis and differentiating xylem. Together, these results suggest that some expansins may be preferentially related to elongation and widening after integrating several spatial, environmental, genetic, and developmental cues.

Circadian rhythms of hydraulic conductance and growth are enhanced by drought and improve plant performance

Circadian rhythms enable plants to anticipate daily environmental variations, resulting in growth oscillations under continuous light. Because plants daily transpire up to 200% of their water content, their water status oscillates from favourable during the night to unfavourable during the day. We show that rhythmic leaf growth under continuous light is observed in plants that experience large alternations of water status during an entrainment period, but is considerably buffered otherwise. Measurements and computer simulations show that this is due to oscillations of plant hydraulic conductance and plasma membrane aquaporin messenger RNA abundance in roots during continuous light. A simulation model suggests that circadian oscillations of root hydraulic conductance contribute to acclimation to water stress by increasing root water uptake, thereby favouring growth and photosynthesis. They have a negative effect in favourable hydraulic conditions. Climate-driven control of root hydraulic conductance therefore improves plant performances in both stressed and non-stressed conditions.

Genetic and Physiological Controls of Growth under Water Deficit

The sensitivity of expansive growth to water deficit has a large genetic variability, higher than that of photosynthesis, and reflects distinct genetic and physiological controls. The sensitivity of expansive growth to water deficit has a large genetic variability, which is higher than that of photosynthesis. It is observed in several species, with some genotypes stopping growth in a relatively wet soil, whereas others continue growing until the lower limit of soil-available water. The responses of growth to soil water deficit and evaporative demand share an appreciable part of their genetic control through the colocation of quantitative trait loci as do the responses of the growth of different organs to water deficit. This result may be caused by common mechanisms of action discussed in this paper (particularly, plant hydraulic properties). We propose that expansive growth, putatively linked to hydraulic processes, determines the sink strength under water deficit, whereas photosynthesis determines source strength. These findings have large consequences for plant modeling under water deficit and for the design of breeding programs.

Analysis of the spatial variability of maize root density. II: Distances between roots

A study was carried out in a maize field in order (i) to study the horizontal variability of the root length per unit volume (Lv), and (ii) to compare two methods of calculation of distances between roots: the first was based on the classical calculation of half the mean distance between neighbouring roots (HMDR), and the second was a direct graphical method carried out on root maps, on 5 superposed horizontal planes. Lv was measured at silking in adjacent, 10−3m3-sized parallelepipeds, and in 2-cm edged cubes taken in non-compacted zone. At a 10−3m3 scale, the distribution of Lv was bimodal in each layer, with one mode in non-compacted zones and the other, which was 20 times lower, below the wheel tracks. The study on a contimetre-sized scale in the non-compacted parts showed a very skewed distribution, with differences between cubes of more than one order of magnitude. The HMDR per layer were smaller than 2 cm to a depth of 60 cm, whereas the direct method showed that an appreciable proportion of points in the soil were at a distance more than 10 times the HMDR from the nearest root. It would therefore have been incorrect to consider, as it is generally assumed in water uptake models, that the maximum distance which water had to travel to the nearest root was the HMDR. Here, root mapping was therefore a better solution than measurement of Lv for characterizing root system as a water sink.

A Common Genetic Determinism for Sensitivities to Soil Water Deficit and Evaporative Demand: Meta-Analysis of Quantitative Trait Loci and Introgression Lines of Maize

Evaporative demand and soil water deficit equally contribute to water stress and to its effect on plant growth. We have compared the genetic architectures of the sensitivities of maize (Zea mays) leaf elongation rate with evaporative demand and soil water deficit. The former was measured via the response to leaf-to-air vapor pressure deficit in well-watered plants, the latter via the response to soil water potential in the absence of evaporative demand. Genetic analyses of each sensitivity were performed over 21 independent experiments with (1) three mapping populations, with temperate or tropical materials, (2) one population resulting from the introgression of a tropical drought-tolerant line in a temperate line, and (3) two introgression libraries genetically independent from mapping populations. A very large genetic variability was observed for both sensitivities. Some lines maintained leaf elongation at very high evaporative demand or water deficit, while others stopped elongation in mild conditions. A complex architecture arose from analyses of mapping populations, with 19 major meta-quantitative trait loci involving strong effects and/or more than one mapping population. A total of 68% of those quantitative trait loci affected sensitivities to both evaporative demand and soil water deficit. In introgressed lines, 73% of the tested genomic regions affected both sensitivities. To our knowledge, this study is the first genetic demonstration that hydraulic processes, which drive the response to evaporative demand, also have a large contribution to the genetic variability of plant growth under water deficit in a large range of genetic material.