Shoot hydraulic impairments induced by root waterlogging: parallels and contrasts with drought.

Soil waterlogging and drought correspond to contrasting water extremes resulting in plant dehydration. Dehydration in response to waterlogging occurs due to impairments to root water transport, but no previous study has addressed whether limitations to water transport occur beyond this organ or whether dehydration alone can explain shoot impairments. Using common bean (Phaseolus vulgaris) as a model species, we report that waterlogging also impairs water transport in leaves and stems. During the very first hours of waterlogging, leaves transiently dehydrated to water potentials close to the turgor loss point, possibly driving rapid stomatal closure and partially explaining the decline in leaf hydraulic conductance. The initial decline in leaf hydraulic conductance (occurring within 24 h), however, surpassed the levels predicted to occur based solely on dehydration. Constraints to leaf water transport resulted in a hydraulic disconnection between leaves and stems, furthering leaf dehydration during waterlogging and after soil drainage. As leaves dehydrated later during waterlogging, leaf embolism initiated and extensive embolism levels amplified leaf damage. The hydraulic disconnection between leaves and stems prevented stem water potentials from declining below the threshold for critical embolism levels in response to waterlogging. This allowed plants to survive waterlogging and soil drainage. In summary, leaf and stem dehydration are central in defining plant impairments in response to waterlogging, thus creating similarities between waterlogging and drought. Yet, our findings point to the existence of additional players (likely chemicals) partially controlling the early declines in leaf hydraulic conductance and contributing to leaf damage during waterlogging.

Gradients in embolism resistance within stems driven by secondary growth in herbs.

The stems of some herbaceous species can undergo basal secondary growth, leading to a continuum in the degree of woodiness along the stem. Whether the formation of secondary growth in the stem base results in differences in embolism resistance between the base and the upper portions of stems is unknown. We assessed the embolism resistance of leaves and the basal and upper portions of stems simultaneously within the same individuals of two divergent herbaceous species that undergo secondary growth in the mature stem bases. The species were Solanum lycopersicum (tomato) and Senecio minimus (fireweed). Basal stem in mature plants of both species displayed advanced secondary growth and greater resistance to embolism than the upper stem. This also resulted in significant vulnerability segmentation between the basal stem and the leaves in both species. Greater embolism resistance in the woodier stem base was found alongside decreases in the pith-to-xylem ratio, increases in the proportion of secondary xylem, and increases in lignin content. We show that there can be considerable variation in embolism resistance across the stem in herbs and that this variation is linked to the degree of secondary growth present. A gradient in embolism resistance across the stem in herbaceous plants could be an adaptation to ensure reproduction or basal resprouting during episodes of drought late in the lifecycle.

Carbon gain is coordinated with enhanced stomatal conductance and hydraulic architecture in coffee plants acclimated to elevated [CO2]: The interplay with irradiance supply.

We recently demonstrated that, under elevated [CO2] (eCa), coffee (Coffea arabica L.) plants grown at high light (HL), but not at low light (LL), display higher stomatal conductance (gs) than at ambient [CO2] (aCa). We then hypothesized that the enhanced gs at eCa/HL, if sustained at the long-term, would lead to adjustments in hydraulic architecture. To test this hypothesis, potted plants of coffee were grown in open-top chambers for 12 months under HL or LL (ca. 9 or 1 mol photons m-2 day-1, respectively); these light treatments were combined with two [CO2] levels (ca. 437 or 705 µmol mol-1, respectively). Under eCa/HL, increased gs was closely accompanied by increases in branch and leaf hydraulic conductances, suggesting a coordinated response between liquid- and vapor-phase water flows throughout the plant. Still under HL, eCa also resulted in increased Huber value (sapwood area-to-total leaf area), sapwood area-to-stem diameter, and root mass-to-total leaf area, thus further improving the water supply to the leaves. Our results demonstrate that Ca is a central player in coffee physiology increasing carbon gain through a close association between stomatal function and an improved hydraulic architecture under HL conditions.

Abscisic acid acts essentially on stomata, not on the xylem, to improve drought resistance in tomato.

Drought resistance is essential for plant production under water-limiting environments. Abscisic acid (ABA) plays a critical role in stomata but its impact on hydraulic function beyond the stomata is far less studied. We selected genotypes differing in their ability to accumulate ABA to investigate its role in drought-induced dysfunction. All genotypes exhibited similar leaf and stem embolism resistance regardless of differences in ABA levels. Their leaf hydraulic resistance was also similar. Differences were only observed between the two extreme genotypes: sitiens (sit; a strong ABA-deficient mutant) and sp12 (a transgenic line that constitutively overaccumulates ABA), where the water potential inducing 50% embolism was 0.25 MPa lower in sp12 than in sit. Maximum stomatal and minimum leaf conductances were considerably lower in plants with higher ABA (wild type [WT] and sp12) than in ABA-deficient mutants. Variations in gas exchange across genotypes were associated with ABA levels and differences in stomatal density and size. The lower water loss in plants with higher ABA meant that lethal water potentials associated with embolism occurred later during drought in sp12 plants, followed by WT, and then by the ABA-deficient mutants. Therefore, the primary pathway by which ABA enhances drought resistance is via declines in water loss, which delays dehydration and hydraulic dysfunction.

Growth and Leaf Gas Exchange Upregulation by Elevated [CO2] Is Light Dependent in Coffee Plants.

Coffee (Coffea arabica L.) plants have been assorted as highly suitable to growth at elevated [CO2] (eCa), although such suitability is hypothesized to decrease under severe shade. We herein examined how the combination of eCa and contrasting irradiance affects growth and photosynthetic performance. Coffee plants were grown in open-top chambers under relatively high light (HL) or low light (LL) (9 or 1 mol photons m-2 day-1, respectively), and aCa or eCa (437 or 705 µmol mol-1, respectively). Most traits were affected by light and CO2, and by their interaction. Relative to aCa, our main findings were (i) a greater stomatal conductance (gs) (only at HL) with decreased diffusive limitations to photosynthesis, (ii) greater gs during HL-to-LL transitions, whereas gs was unresponsive to the LL-to-HL transitions irrespective of [CO2], (iii) greater leaf nitrogen pools (only at HL) and higher photosynthetic nitrogen-use efficiency irrespective of light, (iv) lack of photosynthetic acclimation, and (v) greater biomass partitioning to roots and earlier branching. In summary, eCa improved plant growth and photosynthetic performance. Our novel and timely findings suggest that coffee plants are highly suited for a changing climate characterized by a progressive elevation of [CO2], especially if the light is nonlimiting.

Seeing is believing: what visualising bubbles in the xylem has revealed about plant hydraulic function.

Maintaining water transport in the xylem is critical for vascular plants to grow and survive. The drought-induced accumulation of embolism, when gas enters xylem conduits, causes declines in hydraulic conductance (K ) and is ultimately lethal. Several methods can be used to estimate the degree of embolism in xylem, from measuring K in tissues to directly visualising embolism in conduits. One method allowing a direct quantification of embolised xylem area is the optical vulnerability (OV) technique. This method has been used across different organs and has a high spatial and temporal resolution. Here, we review studies using the OV technique, discuss the main advantages and disadvantages of this method, and summarise key advances arising from its use. Vulnerability curves generated by the OV method are regularly comparable to other methods, including the centrifuge and X-ray microtomography. A major advantage of the OV technique over other methods is that it can be simultaneously used to determine in situ embolism formation in leaves, stems and roots, in species spanning the phylogeny of land plants. The OV method has been used to experimentally investigate the spreading of embolism through xylem networks, associate embolism with downstream tissue death, and observe embolism formation in the field.

Exploring leaf hydraulic traits to predict drought tolerance of Eucalyptus clones.

Ongoing changes in climate, and the consequent mortality of natural and cultivated forests across the globe, highlight the urgent need to understand the plant traits associated with greater tolerance to drought. Here, we aimed at assessing key foliar traits, with a focus on the hydraulic component, that could confer a differential ability to tolerate drought in three commercial hybrids of the most important Eucalyptus species utilized in tropical silviculture: E. urophyla, E. grandis and E. camaldulensis. All genotypes exhibited similar water potential when the 90% stomatal closure (Ψgs90) occurs with Ψgs90 always preceding the start of embolism events. The drought-tolerant hybrid showed a higher leaf resistance to embolism, but the leaf hydraulic efficiency was similar among all genotypes. Other traits presented by the drought-tolerant hybrid were a higher cell wall reinforcement, lower value of osmotic potential at full turgor and greater bulk modulus of elasticity. We also identified that the leaf capacitance after the turgor loss, the ratio between cell wall thickness (t) and lumen breadth (b) ratio (t/b)3, and the minimal conductance might be good proxies for screening drought-tolerant Eucalyptus genotypes. Our findings suggest that xylem resistance to embolism can be an important component of drought tolerance in Eucalyptus in addition to other traits aimed at delaying the development of high tensions in the xylem. Highlight Drought tolerance in tropical Eucalyptus hybrids encompasses a high leaf resistance to embolism and a suite of traits aimed at delaying the development of high tensions in the xylem.

Impaired auxin signaling increases vein and stomatal density but reduces hydraulic efficiency and ultimately net photosynthesis.

Auxins are known to regulate xylem development in plants, but their effects on water transport efficiency are poorly known. Here we used tomato plants with the diageotropica mutation (dgt), which has impaired function of a cyclophilin 1 cis-trans isomerase involved in auxin signaling, and the corresponding wild type (WT) to explore the mutation's effects on plant hydraulics and leaf gas exchange. The xylem of the dgt mutant showed a reduced hydraulically weighted vessel diameter (Dh) (24-43%) and conduit number (25-58%) in petioles and stems, resulting in lower theoretical hydraulic conductivities (Kt); on the other hand, no changes in root Dh and Kt were observed. The measured stem and leaf hydraulic conductances of the dgt mutant were lower (up to 81%), in agreement with the Kt values; however, despite dgt and WT plants showing similar root Dh and Kt, the measured root hydraulic conductance of the dgt mutant was 75% lower. The dgt mutation increased the vein and stomatal density, which could potentially increase photosynthesis. Nevertheless, even though it had the same photosynthetic capacity as WT plants, the dgt mutant showed a photosynthetic rate c. 25% lower, coupled with a stomatal conductance reduction of 52%. These results clearly demonstrate that increases in minor vein and stomatal density only result in higher leaf gas exchange when accompanied by higher hydraulic efficiency.

Xylem embolism spread is largely prevented by interconduit pit membranes until the majority of conduits are gas-filled.

Xylem embolism resistance varies across species influencing drought tolerance, yet little is known about the determinants of the embolism resistance of an individual conduit. Here we conducted an experiment using the optical vulnerability method to test whether individual conduits have a specific water potential threshold for embolism formation and whether pre-existing embolism in neighbouring conduits alters this threshold. Observations were made on a diverse sample of angiosperm and conifer species through a cycle of dehydration, rehydration and subsequent dehydration to death. Upon rehydration after the formation of embolism, no refilling was observed. When little pre-existing embolism was present, xylem conduits had a conserved, individual embolism-resistance threshold that varied across the population of conduits. The consequence of a variable conduit-specific embolism threshold is that a small degree of pre-existing embolism in the xylem results in apparently more resistant xylem in subsequent dehydrations, particularly in angiosperms with vessels. While our results suggest that pit membranes separating xylem conduits are critical for maintaining a conserved individual conduit threshold for embolism when little pre-existing embolism is present, as the percentage of embolized conduits increases, gas movement, local pressure differences and connectivity between conduits increasingly contribute to embolism spread.

Limited plasticity in embolism resistance in response to light in leaves and stems in species with considerable vulnerability segmentation.

Xylem resistance to embolism is a key metric determining plant survival during drought. Yet, we have a limited understanding of the degree of plasticity in vulnerability to embolism. Here, we tested whether light availability influences embolism resistance in leaves and stems. The optical vulnerability method was used to assess stem and leaf resistance to embolism in Phellodendron amurense and Ilex verticillata acclimated to sun and shade microenvironments within the same canopy. In both species, we found considerable segmentation in xylem resistance to embolism between leaves and stems, but only minor acclimation in response to light availability. With the addition of a third species, Betula pubescens, which shows no vulnerability segmentation, we sought to investigate xylem anatomical traits that might correlate with strong vulnerability segmentation. We found a correlation between the area fraction of vessels in the xylem and embolism resistance across species and tissue types. Our results suggest that minimal acclimation of embolism resistance occurs in response to light environment in the same individual and that the degree of vulnerability segmentation between leaves and stems might be determined by the vessel lumen fraction of the xylem.

The interplay between irrigation and fruiting on branch growth and mortality, gas exchange and water relations of coffee trees.

The overall coordination between gas exchanges and plant hydraulics may be affected by soil water availability and source-to-sink relationships. Here we evaluated how branch growth and mortality, leaf gas exchange and metabolism are affected in coffee (Coffea arabica L.) trees by drought and fruiting. Field-grown plants were irrigated or not, and maintained with full or no fruit load. Under mild water deficit, irrigation per se did not significantly impact growth but markedly reduced branch mortality in fruiting trees, despite similar leaf assimilate pools and water status. Fruiting increased net photosynthetic rate in parallel with an enhanced stomatal conductance, particularly in irrigated plants. Mesophyll conductance and maximum RuBisCO carboxylation rate remained unchanged across treatments. The increased stomatal conductance in fruiting trees over nonfruiting ones was unrelated to internal CO2 concentration, foliar abscisic acid (ABA) levels or differential ABA sensitivity. However, stomatal conductance was associated with higher stomatal density, lower stomatal sensitivity to vapor pressure deficit, and higher leaf hydraulic conductance and capacitance. Increased leaf transpiration rate in fruiting trees was supported by coordinated alterations in plant hydraulics, which explained the maintenance of plant water status. Finally, by preventing branch mortality, irrigation can mitigate biennial production fluctuations and improve the sustainability of coffee plantations.

Osmotic adjustment and hormonal regulation of stomatal responses to vapour pressure deficit in sunflower.

Dynamic variation of the stomatal pore in response to changes in leaf-air vapour pressure difference (VPD) constitutes a critical regulation of daytime gas exchange. The stomatal response to VPD has been associated with both foliage abscisic acid (ABA) and leaf water potential (Ψ  l ); however, causation remains a matter of debate. Here, we seek to separate hydraulic and hormonal control of stomatal aperture by manipulating the osmotic potential of sunflower leaves. In addition, we test whether stomatal responses to VPD in an ABA-deficient mutant (w-1) of sunflower are similar to the wild type. Stomatal apertures during VPD transitions were closely linked with foliage ABA levels in sunflower plants with contrasting osmotic potentials. In addition, we observed that the inability to synthesize ABA at high VPD in w-1 plants was associated with no dynamic or steady-state stomatal response to VPD. These results for sunflower are consistent with a hormonal, ABA-mediated stomatal responses to VPD rather than a hydraulic-driven stomatal response to VPD.

Modified Atmosphere Does Not Reduce the Efficacy of Phytosanitary Irradiation Doses Recommended for Tephritid Fruit Flies.

Phytosanitary irradiation (PI) has been successfully used to disinfest fresh commodities and facilitate international agricultural trade. Critical aspects that may reduce PI efficacy must be considered to ensure the consistency and effectiveness of approved treatment schedules. One factor that can potentially reduce PI efficacy is irradiation under low oxygen conditions. This factor is particularly important because storage and packaging of horticultural commodities under low oxygen levels constitute practices widely used to preserve their quality and extend their shelf life. Hence, international organizations and regulatory agencies have considered the uncertainties regarding the efficacy of PI doses for insects infesting fresh commodities stored under low oxygen levels as a rationale for restricting PI application under modified atmosphere. Our research examines the extent to which low oxygen treatments can reduce the efficacy of phytosanitary irradiation for tephritids naturally infesting fruits. The effects of normoxia (21% O2), hypoxia (~5% O2), and severe hypoxia (< 0.5% O2) on radiation sensitivity of third instars of Anastrepha fraterculus (sensu lato), A. ludens (Loew), Bactrocera dorsalis (Hendel), and Ceratitis capitata (Wiedemann) were evaluated and compared at several gamma radiation doses. Our findings suggest that, compared to normoxia, hypoxic and severe-hypoxic conditioning before and during irradiation can increase adult emergence and contribute to advancement of larval development of tephritid fruit flies only at low radiation doses that are not used as phytosanitary treatments. With phytosanitary irradiation doses approved internationally for several tephritids, low oxygen treatments applied before and during irradiation did not increase the emergence rates of any fruit fly species evaluated, and all treated insects died as coarctate larvae. Thus, the findings of our research support a re-evaluation of restrictions related to phytosanitary irradiation application under modified atmospheres targeting tephritid fruit flies.

Drought-induced lacuna formation in the stem causes hydraulic conductance to decline before xylem embolism in Selaginella.

Lycophytes are the earliest diverging extant lineage of vascular plants, sister to all other vascular plants. Given that most species are adapted to ever-wet environments, it has been hypothesized that lycophytes, and by extension the common ancestor of all vascular plants, have few adaptations to drought. We investigated the responses to drought of key fitness-related traits such as stomatal regulation, shoot hydraulic conductance (Kshoot ) and stem xylem embolism resistance in Selaginella haematodes and S. pulcherrima, both native to tropical understory. During drought stomata in both species were found to close before declines in Kshoot , with a 50% loss of Kshoot occurring at -1.7 and -2.5 MPa in S. haematodes and S. pulcherrima, respectively. Direct observational methods revealed that the xylem of both species was resistant to embolism formation, with 50% of embolized xylem area occurring at -3.0 and -4.6 MPa in S. haematodes and S. pulcherrima, respectively. X-ray microcomputed tomography images of stems revealed that the decline in Kshoot occurred with the formation of an air-filled lacuna, disconnecting the central vascular cylinder from the cortex. We propose that embolism-resistant xylem and large capacitance, provided by collapsing inner cortical cells, is essential for Selaginella survival during water deficit.

Relative Tolerance of Three Morphotypes of the Anastrepha fraterculus Complex (Diptera: Tephritidae) to Cold Phytosanitary Treatment.

The Anastrepha fraterculus (Wiedemann) complex is currently comprised of at least eight morphotypes, including several that are likely to be described as new species. It is critical to evaluate whether the morphotypes differ in tolerance to phytosanitary treatments. Temperatures from 0 to 3°C are used as a phytosanitary treatment for some commodities exported from the region and at risk of infestation by the A. fraterculus complex. Description of A. fraterculus morphotypes as new species could result in the annulation of phytosanitary treatment schedules for the new species. This study compared the relative cold tolerance of five populations from three morphotypes of the A. fraterculus complex: Andean, Peruvian, and Brazilian-1. Both a laboratory and wild strain of the Brazilian-1 morphotype were studied. Differences in mortality of third instars of the five A. fraterculus populations reared on nectarines were observed only with short treatment durations at temperatures ranging from 1.38 ± 0.04°C to 1.51 ± 0.08°C (mean ± SEM). Estimated times to achieve the LT99.99682 (probit 9) showed that Brazilian-1 wild, Brazilian-1 laboratory, and Cusco population were the most cold tolerant, followed by Andean and Peruvian, the least cold tolerant morphotype (i.e., Brazilian-1 wild = Brazilian-1 laboratory = Cusco population > Andean > Peruvian). These findings suggest that the current cold treatment schedules of 15 d at ≤ 1.11°C and 17 d at ≤ 1.67°C can be applied as cold treatments to any potential new species that may arise from the A. fraterculus complex.