A four- to seven-fold enhancement of leaf hydraulic conductance by light has been reported in three temperate tree species. The enhancement occurs in the liquid-flow pathway between the petiole and the site of water evaporation. The enhancement occurs within 1 h, and dissipates in darkness over a period of 1 to 10 h depending on species. Here we report light-induced enhancement of leaf hydraulic conductance in a fourth species, bur oak (Quercus macrocarpa Michx.), the dependence of the effect on light flux and color, its absence in leaves of seedlings, and the impact on the response of leaf vein severance and several metabolic inhibitors. The light response of leaf hydraulic conductance approached saturation at a photosynthetic photon flux of 150 mumol m(-2) s(-1). Hydraulic enhancement was greater in response to blue and green light than to visible radiation of longer wavelengths, although at the same irradiance, the response to white light was greater than to light of any single color. Atrazine (a photosystem II inhibitor), fusicoccin (which stimulates plasma membrane-bound H(+)-ATPase) and HgCl(2) (an aquaporin blocker) reduced the light response of leaf lamina hydraulic conductance. When 2-mercaptoethanol was added following mercury treatment, the light response was totally suppressed. Our results are consistent with the notion that the effect of light on leaf lamina hydraulic conductance is controlled by factors acting outside the leaf veins, possibly through light-induced changes in membrane permeability of either mesophyll or bundle sheath cells, or both.
It has previously been shown that hydraulic conductance in bur oak leaves (Quercus macrocarpa Michx.), measured with the high pressure flow meter technique (HPFM), can significantly increase within 30 min following exposure to high irradiance. The present study investigated whether this increase could be explained by an increase in the cell-to-cell pathway and whether the response is linked to changes in the transcript level corresponding to aquaporin genes. Four cDNA sequences showing high similarity to members of the aquaporin gene family from other plant species were characterized from bur oak leaves and the expression levels of these cDNA sequences were examined in leaves by quantitative real-time PCR (QRT-PCR). No change was found in the relative transcript abundance corresponding to these four putative aquaporin genes in leaves with light-induced high hydraulic conductance (exposed to high irradiance) compared to leaves with low hydraulic conductance (exposed to low irradiance). However, in sun leaves that were exposed to different light levels prior to leaf collection (full sunlight, shade, and covered with aluminium foil for 16 h), the relative transcript levels of two of the putative aquaporin genes increased several-fold in shaded leaves compared to the sun-exposed or covered leaves. When the leaves were pressure-infiltrated with the apoplastic tracer dye trisodium 3-hydroxy-5,8,10-pyrenetrisulphonate (PTS3, 0.02%), there was no change in the PTS3 concentration of leaf exudates collected in ambient light or in high irradiance, but there was a small apoplastic acidification. There was also no change in PTS3 concentration between the leaves infiltrated under high irradiance with 0.02% PTS3 or with 0.1 mM HgCl2 in 0.02% PTS3. The results suggest that the putative aquaporin genes that were identified in the present study probably do not play a role in the light responses of hydraulic conductance at the transcript level, but they may function in regulating water homeostasis in leaves adapted to different light conditions. In addition, it is shown that high irradiance induced changes in the pH of the apoplast and that there does not appear to be a significant shift to the cell-to-cell mediated water transport in bur oak leaves exposed to high irradiance as measured by the apoplastic tracer dye.
American elm (Ulmus americana) seedlings were either non-inoculated or inoculated with Hebeloma crustuliniforme, Laccaria bicolor and a mixture of the two fungi to study the effects of ectomycorrhizal associations on seedling responses to soil compaction and salinity. The seedlings were grown in the greenhouse in pots containing noncompacted (0.4 g cm −3 bulk density) and compacted (0.6 g cm −3 bulk density) soil and subjected to 60 mM NaCl or 0 mM NaCl (control) treatments for 3 weeks. All three fungal inocula had similar effects on the responses of elm seedlings to soil compaction and salt treatment. In non-compacted soil, ectomycorrhizal fungi reduced plant dry weights, root hydraulic conductance, but did not affect leaf hydraulic conductance and net photosynthesis. When treated with 60 mM NaCl, ectomycorrhizal seedlings had severalfold lower leaf concentrations of Na + compared with the non-inoculated plants. Soil compaction reduced Na + leaf concentrations in non-ectomycorrhizal plants and decreased dry weights, gas exchange and root hydraulic conductance. However, in ectomycorrhizal plants, soil compaction had little effect on the leaf Na + concentrations and on other measured growth and physiological parameters. Our results demonstrated that ECM associations could be highly beneficial to plants growing in sites with compacted soil such as urban areas.
The present study investigated leaf water transport properties in trembling aspen (Populus tremuloides) leaves. Leaf lamina hydraulic conductance (K(lam)) and stomatal conductance (g(s)) were drastically suppressed by NaF (a general metabolic inhibitor). In leaves treated with 0.2 mM HgCl(2) (an aquaporin blocker), K(lam) declined by 22% when the leaves were sampled in June but the decline was not significant when the leaves were sampled in August. The leaves sampled in June that transpired 30 mM beta-mercaptoethanol following mercury application showed similar K(lam) as those in control leaves transpiring distilled water. When leaves were pressure-infiltrated with 0.1 mM HgCl(2), K(lam) significantly declined by 25%. Atrazine (a photosystem II inhibitor) drastically reduced leaf net CO(2) uptake by the leaves from seedlings and mature trees but did not have any effect on K(lam) regardless of the irradiance at the leaf level during the K(lam) measurements. When PTS(3) (trisodium 3-hydroxy-5,8,10-pyrenetrisulphonate) apoplastic tracer was pressure-infiltrated inside the leaves, its concentration in the leaf exudates did not change from ambient light to high irradiance treatment and declined in the presence of HgCl(2) in the treatment solution. Trembling aspen K(lam) appears to be linked to leaf metabolism and is uncoupled from the short-term variations in photosynthesis. Aquaporin-mediated water transport does not appear to constitute the dominant pathway for the pressure-driven water flow in the leaves of trembling aspen trees.
Aspen ( Populus tremuloides Michx.) roots were treated with cycloheximide, a protein synthesis inhibitor, to examine the role of protein synthesis in root water transport and plant water relations. Within less than 30 min following root application, cycloheximide inhibited steady-state root water flow rates and 1 h after the application of 1 m M cycloheximide, root hydraulic conductivity had decreased by 85% compared with control roots. However, stomatal conductance showed a significant inhibition only after 2 h following cycloheximide treatment. The reduction in root hydraulic conductivity was accompanied by an almost three-fold increase in the apoplastic water flow ratio as determined by the trisodium 3-hydroxy-5,8,10-pyrenesulphonate tracer dye. Cycloheximide-treated roots showed a decrease in the immunostaining intensity of a 32 kDa microsomal protein band that immunoreacted with the AnthPIP1; 1 antibody suggesting a decrease in the membrane aquaporin expression. These changes occurred without severe metabolic disruptions as measured by root respiration. The results point to the importance of proteinmediated transport in roots and the rapidity of response suggests that protein synthesis may be used as a principal regulatory mechanism in root water transport in aspen.
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