Diurnal cycles of embolism formation and repair in petioles of grapevine (Vitis vinifera cv. Chasselas).

The impact of water deficit on stomatal conductance (g(s)), petiole hydraulic conductance (K(petiole)), and vulnerability to cavitation (PLC, percentage loss of hydraulic conductivity) in leaf petioles has been observed on field-grown vines (Vitis vinifera L. cv. Chasselas). Petioles were highly vulnerable to cavitation, with a 50% loss of hydraulic conductivity at a stem xylem water potential (Ψ(x)) of -0.95 MPa, and up to 90% loss of conductivity at a Ψ(x) of -1.5 MPa. K(petiole) described a daily cycle, decreasing during the day as water stress and evapotranspiration increased, then rising again in the early evening up to the previous morning's K(petiole) levels. In water-stressed vines, PLC increased sharply during the daytime and reached maximum values (70-90%) in the middle of the afternoon. Embolism repair occurred in petioles from the end of the day through the night. Indeed, PLC decreased in darkness in water-stressed vines. PLC variation in irrigated plants showed the same tendency, but with a smaller amplitude. The Chasselas cultivar appears to develop hydraulic segmentation, in which petiole cavitation plays an important role as a 'hydraulic fuse', thereby limiting leaf transpiration and the propagation of embolism and preserving the integrity of other organs (shoots and roots) during water stress. In the present study, progressive stomatal closure responded to a decrease in K(petiole) and an increase in cavitation events. Almost total closure of stomata (90%) was measured when PLC in petioles reached >90%.

Limitation of the Cavitron technique by conifer pit aspiration.

The Cavitron technique facilitates time and material saving for vulnerability analysis. The use of rotors with small diameters leads to high water pressure gradients (DeltaP) across samples, which may cause pit aspiration in conifers. In this study, the effect of pit aspiration on Cavitron measurements was analysed and a modified 'conifer method' was tested which avoids critical (i.e. pit aspiration inducing) DeltaP. Four conifer species were used (Juniperus communis, Picea abies, Pinus sylvestris, and Larix decidua) for vulnerability analysis based on the standard Cavitron technique and the conifer method. In addition, DeltaP thresholds for pit aspiration were determined and water extraction curves were constructed. Vulnerability curves obtained with the standard method showed generally a less negative P for the induction of embolism than curves of the conifer method. Differences were species-specific with the smallest effects in Juniperus. Larix showed the most pronounced shifts in P(50) (pressure at 50% loss of conductivity) between the standard (-1.5 MPa) and the conifer (-3.5 MPa) methods. Pit aspiration occurred at the lowest DeltaP in Larix and at the highest in Juniperus. Accordingly, at a spinning velocity inducing P(50), DeltaP caused only a 4% loss of conductivity induced by pit aspiration in Juniperus, but about 60% in Larix. Water extraction curves were similar to vulnerability curves indicating that spinning itself did not affect pits. Conifer pit aspiration can have major influences on Cavitron measurements and lead to an overestimation of vulnerability thresholds when a small rotor is used. Thus, the conifer method presented here enables correct vulnerability analysis by avoiding artificial conductivity losses.

Carbohydrate storage in wood and bark of rubber trees submitted to different level of C demand induced by latex tapping.

When the current level of carbohydrates produced by photosynthesis is not enough to meet the C demand for maintenance, growth or metabolism, trees use stored carbohydrates. In rubber trees (Hevea brasiliensis Muell. Arg.), however, a previous study (Silpi U., A. Lacointe, P. Kasemsap, S. Thanisawanyangkura, P. Chantuma, E. Gohet, N. Musigamart, A. Clement, T. Améglio and P. Thaler. 2007. Carbohydrate reserves as a competing sink: evidence from tapping the rubber tree. Tree Physiol. 27:881-889) showed that the additional sink created by latex tapping results not in a decrease, but in an increase in the non-structural carbohydrate (NSC) storage in trunk wood. In this study, the response of NSC storage to latex tapping was further investigated to better understand the trade-off between latex regeneration, biomass and storage. Three tapping systems were compared to the untapped Control for 2 years. Soluble sugars and starch were analyzed in bark and wood on both sides of the trunk, from 50 to 200 cm from the ground. The results confirmed over the 2 years that tapped trees stored more NSC, mainly starch, than untapped Control. Moreover, a double cut alternative tapping system, which produced a higher latex yield than conventional systems, led to even higher NSC concentrations. In all tapped trees, the increase in storage occurred together with a reduction in trunk radial growth. This was interpreted as a shift in carbon allocation toward the creation of reserves, at the expense of growth, to cover the increased risk induced by tapping (repeated wounding and loss of C in latex). Starch was lower in bark than in wood, whereas it was the contrary for soluble sugars. The resulting NSC was twice as low and less variable in bark than in wood. Although latex regeneration occurs in the bark, changes related to latex tapping were more marked in wood than in bark. From seasonal dynamics and differences between the two sides of the trunk in response to tapping, we concluded that starch in wood behaved as the long-term reserve compartment at the whole trunk level, whereas starch in bark was a local buffer. Soluble sugars behaved like an intermediate, ready-to-use compartment in both wood and bark. Finally, the dynamics of carbohydrate reserves appears a relevant parameter to assess the long-term performance of latex tapping systems.

Carbohydrate reserves as a competing sink: evidence from tapping rubber trees.

Carbohydrate reserve storage in trees is usually considered a passive function, essentially buffering temporary discrepancies between carbon availability and demand in the annual cycle. Recently, however, the concept has emerged that storage might be a process that competes with other active sinks for assimilate. We tested the validity of this concept in Hevea brasiliensis Müll. Arg. (rubber) trees, a species in which carbon availability can be manipulated by tapping, which induces latex regeneration, a high carbon-cost activity. The annual dynamics of carbohydrate reserves were followed during three situations of decreasing carbon availability: control (no tapping), tapped and tapped with Ethephon stimulation. In untapped control trees, starch and sucrose were the main carbohydrate compounds. Total nonstructural carbohydrates (TNC), particularly starch, were depleted following bud break and re-foliation, resulting in an acropetal gradient of decreasing starch concentration in the stem wood. During the vegetative season, TNC concentration increased. At the end of the vegetative season, there were almost no differences in TNC concentration along the trunk. In tapped trees, the vertical gradient of starch concentration was locally disturbed by the presence of the tapping cut. However, the main effect of tapping was a dramatic increase in TNC concentration, particularly starch, throughout the trunk and in the root. The difference in TNC concentration between tapped and untapped trees was highest when latex production was highest (October); the difference was noticeable even in areas of the trees that are unlikely to be directly involved in latex regeneration, and it was enhanced by Ethephon stimulation, which is known to increase latex metabolism and flow duration. Thus, contrary to what could be expected if reserves serve as a passive buffer, a decrease in carbohydrate availability resulted in a net increase in carbohydrate reserves at the trunk scale. Such behavior supports the view that trees tend to adjust the amount of carbohydrate reserves stored to the level of metabolic demand, at the possible expense of growth.

Stem diameter variations and cold hardiness in walnut trees.

The effect of freezing temperatures on stem diameter was measured in the field and in climatic chambers using linear variable differential transformers (LVDT sensors). In acclimated stems, there was reversible stem shrinkage associated with freeze-thaw cycles. The maximum shrinkage correlated with stem diameter (thickness of the bark). The wood was responsible for only 15% of the shrinkage associated with a freeze event, and experiments with isolated bark showed that connection with the wood was not necessary for most of the freeze-induced shrinkage to occur. Considering the amount of stem shrinkage associated with summer drought in walnut, the amount of contraction of the bark with freezing was actually much less than might be predicted by water relations theory. Reversible stem shrinkage occurred in living tissues, but not in autoclaved tissues. For the latter, swelling was observed with freezing and this swelling could be explained by the bark alone. Similar swelling was observed during September and October for non-acclimated plants. Water was lost with each freeze-thaw cycle starting with the first, and freezing injury of the bark, with discoloration of tissues, was also observed in non-acclimated plants. Given that the diameter fluctuation patterns were dramatically different for acclimated versus non-acclimated plants, and for living versus autoclaved tissues, LVDT sensors could represent a novel, non-invasive approach to testing cold hardiness.

Seasonal variation in xylem pressure of walnut trees: root and stem pressures.

Measurements of air and soil temperatures and xylem pressure were made on 17-year-old orchard trees and on 5-year-old potted trees of walnut (Juglans regia L.). Cooling chambers were used to determine the relationships between temperature and sugar concentration ([glucose] + [fructose] + [sucrose], GFS) and seasonal changes in xylem pressure development. Pressure transducers were attached to twigs of intact plants, root stumps and excised shoots while the potted trees were subjected to various temperature regimes in autumn, winter and spring. Osmolarity and GFS of the xylem sap (apoplast) were measured before and after cooling or warming treatments. In autumn and spring, xylem pressures of up to 160 kPa were closely correlated with soil temperature but were not correlated with GFS in xylem sap. High root pressures were associated with uptake of mineral nutrients from soil, especially nitrate. In autumn and spring, xylem pressures were detected in root stumps as well as in intact plants, but not in excised stems. In contrast, in winter, 83% of the xylem sap osmolarity in both excised stems and intact plants could be accounted for by GFS, and both GFS and osmolarity were inversely proportional to temperature. Plants kept at 1.5 degrees C developed positive xylem pressures up to 35 kPa, xylem sap osmolarities up to 260 mosmol l(-1) and GFS concentrations up to 70 g l(-1). Autumn and spring xylem pressures, which appeared to be of root origin, were about 55% of the theoretical pressures predicted by osmolarity of the xylem sap. In contrast, winter pressures appeared to be of stem origin and were only 7% of the theoretical pressures, perhaps because of a lower stem water content during winter.

A new validation of the Scholander pressure chamber technique based on stem diameter variations.

The Scholander pressure chamber is one of the most widely used techniques for measuring plant water status. However, the technique has been the subject of recent controversies, and its validity awaits new experimental evidence. This paper presents a new test based on the analysis of the dependence on water potential difference (DeltaPsi) of stem diameter variation (DeltaD) in walnut (Juglans regia L.). The correlation between DeltaPsi and DeltaD was established (1) on transpiring potted trees, (2) on dehydrating cut branches, (3) by perfusing the xylem of branch segments with mannitol and sucrose solutions, and (4) by pressurizing segments in a pressure sleeve. The DeltaPsi was respectively assessed with a pressure chamber (1, 2), a freezing point osmometer (3) and an air pressure transducer (4). A single relationship was established between DeltaPsi (ranging from 0 to -2 MPa) and DeltaD for all the experiments. This shows that the measured changes of water potential were correlated to similar modifications of water content in the stems, irrespective of the technique used to induce these changes, and therefore validates the pressure chamber technique and confirms the occurrence of large negative pressures in the xylem of walnut branches.

Winter stem xylem pressure in walnut trees: effects of carbohydrates, cooling and freezing.

Pressure transducers were attached to twigs of orchard trees and potted trees of walnut (Juglans regia L.) to measure winter stem xylem pressures. Experimental potted trees were partially defoliated in the late summer and early autumn to lower the amount of stored carbohydrates. Potted trees were placed in cooling chambers and subjected to various temperature regimes, including freeze-thaw cycles. Xylem pressures were inversely proportional to the previous 48-h air temperature, but positively correlated with the osmolarity of the xylem sap. Defoliated trees had significantly lower concentrations of stored carbohydrates and significantly lower xylem sap osmolarities than controls. Plants kept at 1.5 degrees C developed xylem pressures up to 40 kPa, just 7% of the theoretical osmotic pressure of the xylem sap. However, exposure to low, nonfreezing temperatures followed by freeze-thaw cycles resulted in pressures over 210 kPa, which was 39% of the theoretical osmotic pressure. A simple osmotic model could account for the modest positive winter pressures at low, nonfreezing temperatures, but not for the synergistic effects of freeze-thaw cycles.

Mechanisms of xylem recovery from winter embolism in Fagus sylvatica.

Hydraulic conductivity in the terminal branches of mature beech trees (Fagus sylvatica L.) decreased progressively during winter and recovered in the spring. The objective of this study was to determine the mechanisms involved in recovery. Two periods of recovery were identified. The first recovery of hydraulic conductivity occurred early in the spring, before bud break, and was correlated with the occurrence of positive xylem pressure at the base of the tree trunk. Active refilling of the embolized vessels caused the recovery. The second recovery of hydraulic conductivity occurred after bud break and was correlated with the onset of cambial activity. Formation of new functional vessels, leading to an increase in xylem diameter, was largely responsible for the increase in xylem conductivity. The two mechanisms were complementary: active refilling of embolized vessels occurred mostly in the root and the trunk, whereas formation of new functional vessels occurred mainly in young terminal shoots.

Cryo-scanning electron microscopy observations of vessel content during transpiration in walnut petioles. Facts or artifacts?

The current controversy about the "cohesion-tension" of water ascent in plants arises from the recent cryo-scanning electron microscopy (cryo-SEM) observations of xylem vessels content by Canny and coworkers (1995). On the basis of these observations it has been claimed that vessels were emptying and refilling during active transpiration in direct contradiction to the previous theory. In this study we compared the cryo-SEM data with the standard hydraulic approach on walnut (Juglans regia) petioles. The results of the two techniques were in clear conflict and could not both be right. Cryo-SEM observations of walnut petioles frozen intact on the tree in a bath of liquid nitrogen (LN(2)) suggested that vessel cavitation was occurring and reversing itself on a diurnal basis. Up to 30% of the vessels were embolized at midday. In contrast, the percentage of loss of hydraulic conductance (PLC) of excised petiole segments remained close to 0% throughout the day. To find out which technique was erroneous we first analyzed the possibility that PLC values were rapidly returned to zero when the xylem pressures were released. We used the centrifugal force to measure the xylem conductance of petiole segments exposed to very negative pressures and established the relevance of this technique. We then analyzed the possibility that vessels were becoming partially air-filled when exposed to LN(2). Cryo-SEM observations of petiole segments frozen shortly after their xylem pressure was returned to atmospheric values agreed entirely with the PLC values. We confirmed, with water-filled capillary tubes exposed to a large centrifugal force, that it was not possible to freeze intact their content with LN(2). We concluded that partially air-filled conduits were artifacts of the cryo-SEM technique in our study. We believe that the cryo-SEM observations published recently should probably be reconsidered in the light of our results before they may be used as arguments against the cohesion-tension theory.