Sebastiano Salleo

Refilling embolized xylem conduits: is it a matter of phloem unloading?

Long-distance water transport in plants relies on negative pressures established in continuous water columns in xylem conduits. Water under tension is in a metastable state and is prone to cavitation and embolism, which leads to loss of hydraulic conductance, reduced productivity and eventually plant death. Experimental evidence suggests that plants can repair embolized xylem by pushing water from living vessel-associated cells into the gas-filled conduit lumina. Most surprisingly, embolism refilling is known to occur even when the bulk of still functioning xylem is under tension, a finding that is in seemingly contradiction to basic principles of thermodynamics. This review summarizes our current understanding of xylem refilling processes and speculates that embolism repair under tension can be envisioned as a particular case of phloem unloading, as suggested by several events and components of embolism repair, typically involved in phloem unloading mechanisms. Far from being a challenge to irreversible thermodynamics, embolism refilling is emerging as a finely regulated vital process essential for plant functioning under different environmental stresses.

Phloem as a possible major determinant of rapid cavitation reversal in stems of Laurus nobilis (laurel)

Xylem recovery from embolism was studied in stems of Laurus nobilis L. that were induced to cavitate by combining negative xylem pressures with positive air pressures applied with a pressure collar. Xylem refilling was measured 2 and 20 min and 15 h after air pressure release in January, March and June when increasing percentages of wood parenchyma cells with high starch content (HSC-VAC) were counted (from 0% in January to 87.3% in June). In January, no xylem repair was measured. In June, stems refilled by 75% of previous conductivity loss with a parallel decrease of HSC-VAC. Xylem refilling was tested for stems with phloem either intact or excised by 20 and 50% and with phloem inactivated by girdling stems at both sides of the embolised segment. Stems with 50% of the cortex removed showed some recovery 15 h after embolism. Girdled stems did not recover from embolism and no starch depolymerisation was measured. Girdled stems where a radial mechanical pressure was applied for 20 min after embolism refilled in the same way as stems with intact phloem. Our conclusion is that phloem may export some signal for starch depolymerisation and this, in turn, would drive sugar efflux into embolised conduits with consequent osmotic water flows and refilling.

Hydraulic efficiency of the leaf venation system in sun- and shade-adapted species

We tested the hypothesis that leaf hydraulics is correlated with the light adaptation of different plant species and specifically that the hydraulic resistance of the leaf venation (Rvenation) is lower in sun- than in shade-adapted species. Rvenation was measured in six sun- and six shade-adapted species with a high-pressure flow meter (HPFM). The number of conduits at the proximal third of the midrib was counted and the diameter of the widest conduits together with vein density were measured. Rvenation was higher in shade species than in sun species and it was negatively correlated with the mean diameter of the widest conduits. Maximum leaf conductance to water vapour recorded for the different species was negatively correlated with the corresponding Rvenation. Sun-adapted species coping with the high water demand typical of sunny habitats appeared to have developed a highly efficient conducting system to supply living mesophyll cells with water. In contrast, species adapted to shady habitats showed higher Rvenation values according to their lower need for investment of carbon into producing wide conduits in the leaf.

Effects of defoliation caused by the leaf miner Cameraria ohridella on wood production and efficiency in Aesculus hippocastanum growing in north-eastern Italy

The leaf miner Cameraria ohridella causes premature defoliation of Aesculus hippocastanum trees. Repeated defoliation has been reported to cause decrease in radial growth of trees and a progressive decline due to reduced production and allocation of photosynthates. Our study represents an attempt to estimate the impact of C. ohridella on annual wood increments and the hydraulic properties of the wood as well as on the dry mass of seeds. Twenty-two adult horse chestnut trees were selected, four of which had been chemically treated to prevent attack (controls). All other trees were heavily infested. The ground cover (GC) of each tree, measured from monthly hemispherical photographs, revealed that infested trees were completely defoliated in September and the slope of the GC-to-measurement dates relationship (named GC decrease index) was positively related to the number of mines per leaf. Anatomical observations showed that infested trees produced more wood per year than controls through more false rings with wider xylem conduits and, hence, with higher conductive area and theoretical flow. In fact, the theoretical flow was positively related to the defoliation intensity. In contrast, the allocation of photosynthates to seeds was strongly reduced in infested trees with respect to controls (up to 50% less). The hypothesis was advanced that horse chestnut trees reacted to C. ohridella attacks by increasing the hydraulic efficiency of the wood, thus ameliorating the water and nutrient supply to leaves between the spring and mid-summer and, therefore, compensating, at least partly, the reduced leaf lifespan.

Sclerophylly: Evolutionary advantage or mere epiphenomenon?

ABSTRACT The possible functional significance of sclerophylly (hard and coriaceous leaves) is discussed on the basis of different interpretations reported in old and recent literature. These, have been schematically divided into four groups of hypotheses, i.e. the “water conservation˚d hypothesis, the “resistance to negative turgour pressures˚d hypothesis, the “nutritional˚d hypothesis and the “protective˚d hypothesis. In particular, a section is devoted to discussing the first and second hypotheses in terms of relationships of sclerophylly to drought resistance strategies. Data reported in the literature lead to the conclusion that the sclerophyllous habitus per se seems not to be related to drought resistance of plants as previously thought. Nor do clear-cut relationships appear to exist between leaf sclerification and living leaf cells with thicker walls, leading to a high modulus of elasticity (ε), the latter known to be of importance for generating low leaf water potentials, i.e. higher driving forces for water uptake. Because leaf sclerification has proved to be related to soil nutrient deficiency, this leaf habitus can be considered as a mere epiphenomenon of nutrient stress. On the other hand, the selective advantage of sclerophylly as protection against herbivore attacks, though not demonstrated in different environments, seems to be of great interest and deserves further studies.

Impact of the leaf miner Cameraria ohridella on photosynthesis, water relations and hydraulics of Aesculus hippocastanum leaves

The mining of leaves of Aesculus hippocastanum caused by the larvae of Cameraria ohridella leads to precocious defoliation of trees. Damage to plant productivity was estimated in terms of the photosynthetic performance as well as of leaf water relations and hydraulics of increasingly mined leaves from infested plants in comparison with the same variables measured in non-mined leaves (controls). Electron microscopy and photosynthesis measurements revealed that chloroplasts within the green portions of mined leaves were still intact and photosynthesis of these areas was close to that of non-mined leaves, i.e. damage to functional integrity of the photosynthetic system did not extend beyond the mines. Stomata below the mines were functional as they maintained their physiological kinetics but most chloroplasts in the spongy parenchyma below the mines were degraded so that a 1:1 relationship existed between photosynthesis loss and loss of leaf green areas. Leaf conductance to water vapour and transpiration rate were 60% lower in mined leaf areas but equal to controls in green portions of mined leaves. Leaf water potential was insensitive to the amount of mined leaf area and so was leaf hydraulic conductance. Anatomical observations of leaf minor veins revealed that they were structurally and functionally intact even in leaves with 90% mined surface area. Our conclusion was that the actual damage to A. hippocastanum plants in terms of loss of photosynthates and water and nutrient transport was less than that visually estimated in recent studies.

Resistance to water flow through leaves of Coffea arabica is dominated by extra-vascular tissues

The leaf hydraulic conductance (Kleaf) of Coffea arabica L. was measured for shoots exposed to non-lethal temperature stress or to a freeze-thaw cycle, and compared with Kleaf of non-stressed samples (controls). Exposure to temperatures below 6°C for 1 h caused measurable damage to the functional integrity of cell membranes as shown by increased membrane leakiness to electrolytes. A 1 : 1 relationship was found to exist between relative electrolyte leakage and relative Kleaf suggesting that membrane damage caused Kleaf to increase. Low temperatures did not cause membrane disruption as shown by the comparison of chilled samples with frozen-thawed ones. In frozen leaves, membranes were extensively disrupted and both electrolyte leakiness and Kleaf increased 5-fold. Low temperatures did not induce alterations of the hydraulic properties of the leaf vascular system, as revealed by measurements of Kleaf after up to 500 cuttings of minor veins were made in the leaf blade of control and chilled leaves. Calculations showed that 62-75% of leaf hydraulic resistance resided in the extra-vascular water pathway. Results are discussed within the frame work of our current understanding of leaf hydraulic architecture as well as in terms of plant adaptation to extremes in temperature.

Leafminers help us understand leaf hydraulic design

Leaf hydraulics of Aesculus hippocastanum L. were measured over the growing season and during extensive leaf mining by the larvae of an invasive moth (Cameraria ohridella Deschka et Dimic) that specifically destroy the palisade tissue. Leaves showed seasonal changes in hydraulic resistance (R(lamina)) which were related to ontogeny. After leaf expansion was complete, the hydraulic resistance of leaves and the partitioning of resistances between vascular and extra-vascular compartments remained unchanged despite extensive disruption of the palisade by leafminers (up to 50%). This finding suggests that water flow from the petiole to the evaporation sites might not directly involve the palisade cells. The analysis of the temperature dependence of R(lamina) in terms of Q(10) revealed that at least one transmembrane step was involved in water transport outside the leaf vasculature. Anatomical analysis suggested that this symplastic step may be located at the bundle sheath where the apoplast is interrupted by hydrophobic thickening of cell walls. Our findings offer some support to the view of a compartmentalization of leaves into well-organized water pools so that the transpiration stream would involve veins, bundle sheath and spongy parenchyma, while the palisade tissue would be largely by-passed with the possible advantage of protecting cells from short-term fluctuations in water status.

Changes in stem and leaf hydraulics preceding leaf shedding in Castanea sativa L.

This paper describes changes in leaf water status and in stem, petiole and leaf blade hydraulics preceding leaf senescence and shedding in Castanea sativa L. (chestnut). Measurements of maximum diurnal leaf conductance to water vapour (gL), minimum water potential (ψL), hydraulic conductance per unit leaf surface area of stems (KSL), petioles (KPL) and leaf blades (KLL) and number of functional conduits and inside diameter distribution were performed in June, September and October 1999. In September, still green leaves had undergone some dehydration as indicated by decreased gL (by 75 %) and ψL with respect to June. In the same time, KSL decreased by 88 %, while KPL and KLL decreased by 50 % and 20 % of the conduits of stems and 10 % of the petioles (all belonging to the widest diameter range) were no longer functioning, causing a decrease in the theoretical flow by 82 % in stems and 27 % in petioles. Stem xylem blockage was apparently due to tyloses growing into conduits. We advance the hypothesis that the entire process of leaf shedding and winter rest may be initiated by extensive stem embolism occurring during the summer.

Short-term effects of potassium fertilization on the hydraulic conductance of Laurus nobilis L.

This study reports experimental evidence on the effect of short-term potassium fertilization on potassium uptake, tissue concentration and hydraulic conductance of pot-grown laurel plants. Potassium uptake and loading into the xylem of laurel seedlings increased within 24 h after fertilization. Potassium was not accumulated in roots and leaves, but the [K(+)] of xylem sap was 80% higher in fertilized plants (+K) than in potassium-starved plants (-K), as a likely result of recirculation between xylem and phloem. Increased xylem sap [K(+)] resulted in a 45% increase in transpiration rate, a 30% increase in plant hydraulic conductance (K(plant)) and a 120% increase in leaf-specific conductivity of the shoot (k(shoot)). We suggest that this increase was due to ion-mediated up-regulation of xylem hydraulics, possibly caused by the interaction of potassium ions with the pectic matrix of intervessel pits. The enhancement of hydraulic conductance following short-term potassium fertilization is a phenomenon that can be of advantage to plants for maintaining cell turgor, stomatal aperture and gas exchange rates under moderate drought stress. Our data provide additional support for the important role of potassium nutrition in agriculture and forestry.