Quantifying determinants of ozone detoxification by apoplastic ascorbate in peach (Prunus persica) leaves using a model of ozone transport and reaction.

Ascorbate in leaf apoplast (ASCapo ) reacts with ozone (O3 ) and thereby reduces O3 flux reaching plasmalemma (Fpl ). Some studies have shown significant protection of cells from O3 by ASCapo , while others have questioned its efficacy. Hypothesizing that the protection by ASCapo depends on other variables, we quantified determinants of O3 detoxification with a model of O3 transport and reaction in apoplast. The model determines ascorbic acid concentration in apoplast (AAapo ) using measured values of O3 concentration (co ), leaf tissue ascorbic acid concentration (AAleaf ), cell wall thickness (L3 ), apoplastic pH (pHapo ), and stomatal conductance (Gsw ). We compared the measured and model-estimated AAapo in leaves of peach (Prunus persica) grown in open-top chambers under non-filtered air (NF) and elevated (EO3 : NF + 80 ppb) O3 concentrations. The estimated AAapo in individual leaves agreed well with the measured values (R2  = .91). Analyses of the simulation results yielded the following findings: (a) The efficacy of O3 reduction with ASCapo as quantified by fractional reduction (ϕ3 ) of O3 flux at the surface of plasmalemma (Fpl ) was lowered from 70% in NF to 40% in EO3 due to the reduction of L3 . The EO3 reduced AAapo , but the lower Gsw and L3 in EO3 increased AAapo resulting in no significant change in AAapo due to EO3 . ϕ3 can be calculated with measured values of AAapo and L3 , and Fpl can be estimated with the measurement-based ϕ3 . (b) When c0 is increased, Fpl increased curvilinearly with the increase of Fst : nominal O3 flux via stomatal diffusion, exhibiting apparent threshold on Fst . The deviation of Fpl from Fst became greater when L3 , pHapo , and AAleaf were increased. The quantification of ϕ3 and Fpl using leaf traits shall facilitate the understanding of the mechanisms of differential plant sensitivity to O3 and improve quantification of the O3 impacts on plants.

Overcoming the difficulties in collecting apoplastic fluid from rice leaves by the infiltration-centrifugation method.

Physiological and biochemical studies on the leaf apoplast have been facilitated by the use of the infiltration-centrifugation technique to collect intercellular washing fluid (IWF). However, this technique has been difficult to implement in rice (Oryza sativa L.) for various reasons. We compared the collection efficiency of leaf IWF between two types of rice varieties (Indica and Japonica), as well as between rice and other species (spinach, snap bean and wheat). Although the extraction of IWF in most species took only 2-3 min, it took up to 35 min in rice. The difficulty in infiltration with rice was ascribed to the small stomatal aperture and hydrophobicity of the leaves. In this study, we have established an improved method for collecting IWF and determining the apoplastic air and water volumes in rice leaves. We have shortened the infiltration time to 8 min via the following improvements: (i) infiltration under outdoor shade in the daytime to prevent stomatal closure and a rise in temperature of the infiltration medium; (ii) soaking of leaves in a surfactant solution to decrease the leaf hydrophobicity; and (iii) continuous pressurization using a sealant injector to facilitate the infiltration. The rapid collection of IWF achieved using this technique will facilitate study of the leaf apoplast in rice.

Apoplastic ascorbate contributes to the differential ozone sensitivity in two varieties of winter wheat under fully open-air field conditions.

We studied leaf apoplastic ascorbates in relation to ozone (O(3)) sensitivity in two winter wheat (Triticum aestivum L.) varieties: Yangfumai 2 (Y2) and Yangmai 16 (Y16). The plants were exposed to elevated O(3) concentration 27% higher than the ambient O(3) concentration in a fully open-air field from tillering stage until final maturity. The less sensitive variety Y16 had higher concentration of reduced ascorbate in the apoplast and leaf tissue by 33.5% and 12.0%, respectively, than those in the more sensitive variety Y2, whereas no varietal difference was detected in the decline of reduced ascorbate concentration in response to elevated O(3). No effects of O(3) or variety were detected in either oxidized ascorbate or the redox state of ascorbate in the apoplast and leaf tissue. The lower ascorbate concentrations in both apoplast and leaf tissue should have contributed to the higher O(3) sensitivity in variety Y2.