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Agnes A. Flores-Nimedez, Paul H. Li, and Charles C. Shin

GLK-8903, an experimental product whose main ingredient is produced by hydrogenation of a primary alcohol extracted from plants, showed significant potential in protecting bean (Phaseolus vulgaris L.) plants from chilling injury. The GLK-8903 protection mechanism was assessed by examining several physiological and biochemical responses. The decline in leaf water potential and the increase in osmotic potential caused by chilling exposure to 4C (day/night) were minimized by the application of GLK-8903. Chilling causes an increase in electrolyte leakage, an indication of chilling injury of the plasma membrane. Increased electrolyte leakage was reduced significantly in the GLK-8903-treated plants during chilling. This minimized leakage may be due to less damage of the plasma membrane. Plasmolysis and deplasmolysis studies of the epidermal cells suggest that GLK-8903 is able to reduce the plasma membrane perturbation in the chilling environment, as evident by: 1) the lower permeability coefficient to urea at 4C, and 2) the swelling of protoplasts in the cells of untreated tissues after chilling exposure with no swelling of the protoplast being observed in the GLK-8903-treated cells. Malondialdehyde (MDA), a product of lipid peroxidation, increased more in untreated controls than in treated plants exposed to 4C. Plasma membrane ATPase activity decreased less in GLK-8903-treated plants than in untreated controls after 3 days at 4C. The mechanism of GLK-8903-alleviated chilling injury is discussed.

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Eckhard Grimm and Moritz Knoche

used microscopy and a plasmolysis technique to limit our analyses just to the epidermal cell layer. The same values for the flesh were measured conventionally using osmometry. Materials and Methods Plant material. Fruit of the sweet cherry cultivars

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Albert H. Markhart III and Mark S. Harper

Leaves on cut stems of commercially grown Rosa hybrida cv. Kardinal placed in preservative solutions containing sucrose developed necrotic dry patches that began interveinally and progressed toward the major veins until the entire leaf was dehydrated. Ultrastructural observations of initial damage showed disorganized protoplasm and plasmolyzed cells. Leaves on cut stems pretreated with abscisic acid for 24 hours and transferred to preservative solution containing sucrose remained healthy. We propose that sucrose accumulates in the mesophyll cell wall, thus decreasing apoplastic osmotic potential, leading to cell collapse and tissue death.

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ZhaoSen Xie, Charles F. Forney, WenPing Xu, and ShiPing Wang

control fruit, showing plasmolysis in CC. ( C ) A mature SE/CC complex in a fruit produced under root restriction showing more serious plasmolysis in CC. ( D ) A mature SE/CC complex in a fruit produced under root restriction showing dense cytoplasm with

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Marie-Therese Charles, Alain Goulet, Francois Castaigne, and Joseph Arul

Hormic dose of ultraviolet light (3.7 kJ•m-2) induced disease resistance in tomato fruit. The biochemical nature of induced resistance by UV light was investigated by histochemical techniques. Ultraviolet light induced plasmolysis of the epicarp and few mesocarp cell layers, and collapse of these cell layers led to the formation of cell wall stacking zone (CWSZ). The treatment also stimulated the biosynthesis of phenolic compounds (Prussian blue reaction) in the epicarp and mesocarp cells. Biochemical reinforcement of the cell wall through lignification (Maule test) and suberization (berberine fluorescence) was also induced. These responses originating from the activation of phenylpropanoid path were principally localized in the CWSZ and were induced before inoculation by B. cinerea. The intensity of these responses was significantly increased in UV-treated tissue in response to infection. These responses were also induced in the inoculated control tissue but were either less substantial (phenolics, lignification, and suberization) or delayed.

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B.R. Bondada, R Romero-Aranda, J. Syvertsen, and L. Albrigo

Foliar applications of urea-nitrogen are widely used to alleviate N deficiencies in citrus; however, improper applications can cause serious foliar burn and loss of active green leaf area. Light (LM), transmission (TEM), and scanning (SEM) electron microscopy were used to characterize anatomical and ultrastructural details of foliar burn in citrus. LM examination of the burned leaf area showed collapsed adaxial and abaxial epidermal cells and plasmolysis of mesophyll cells that created large intercellular spaces. SEM showed wrinkling of both the adaxial and abaxial epidermal cells. TEM revealed cytoplasmic vacuolation, disruption of cellular membrane, degradation of grana, and appearance of large plastoglobuli, implying loss of physiological activity. In contrast, control leaves had turgid adaxial and abaxial epidermal cells and compact mesophyll cells with few intercellular air spaces.

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J.G. Luza, R. van Gorsel, V.S. Polito, and A.A. Kader

Fruits of mid- (`O'Henry'), late (`Airtime'), and extra-late-season (`Autumn Gem') peach [Prunus persica (L.) Batsch] cultivars were examined for changes in cell wall structure and cytochemistry that accompany the onset of mealiness and leatheriness of the mesocarp due to chilling injury. The peaches were stored at 10C for up to 18 days or at SC for up to 29 days. Plastic-embedded sections were stained by the Schiff's-periodic acid reaction, Calcofluor white MR2, and Coriphosphine to demonstrate total insoluble carbohydrates, ß-1,4 glucans, and pectins, respectively. Mealiness was characterized by separation of mesocarp parenchyma cells leading to increased intercellular spaces and accumulation of pectic substances in the intercellular matrix. Little structural change was apparent in the cellulosic component of the cell walls of these fruits. In leathery peaches, the mesocarp parenchyma cells collapsed, intercellular space continued to increase, and pectin-positive staining in the intercellular matrix increased greatly. In addition, the component of the cell walls that stained positively for ß-1,4 glucans became thickened relative to freshly harvested or mealy fruit. At the ultrastructural level, dissolution of the middle lamella, cell separation, irregular thickening of the primary wall, and plasmolysis of the mesocarp parenchyma cells were seen as internal breakdown progressed.

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Edwin J. Reidel, Lailiang Cheng, and Robert Turgeon

Sorbitol is the predominant phloem-translocated carbohydrate in apple. The pathway—either apoplastic or symplastic—of sugar transport from photosynthetic cells to the phloem is not established. Furthermore, the presence of absence of phloem loading has not been tested. This study characterized the morphology and physiology of sugar movement to the phloem in apple leaves. An electron micrographic survey of apple leaf minor vein morphology was performed. Plasmodesmata were abundant and found at the interfaces of each cell type from mesophyll to sieve elements, indicating a symplastic sugar pathway. We also tested for a phloem loading mechanism. First, 14C-labeled sorbitol and sucrose were introduced exogenously to leaf discs to determine if they are loaded into veins from the apoplast. Although leaf discs floated on a solution containing either sugar actively accumulated label, the labeling pattern was diffuse, with no accumulation in minor veins. The addition of the sulfhydryl reagent PCMBS to the leaf disc assay inhibited sugar uptake. We also attempted plasmolysis of apple leaf sections to measure the solute concentration difference between photosynthetic mesophyll cells and cells of the minor vein phloem. Apple leaf pieces fixed in a solution containing 1.5 mol/kg osmoticum did not plasmolyze. We conclude that although active uptake of both sorbitol and sucrose takes place in apple leaves, apoplastic phloem-loading is absent. Considering the high sugar concentration and the symplastic connectivity among leaf cell types, we propose that sugars are instead enter the phloem after moving down—rather than against—a concentration gradient.

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Agnes A. Flores-Nimedez, Paul H. Li, and Charles C. Shin

Protection mechanism of a new compound, coded as GLK-8903, from chilling injury in bean plants was assessed by measuring several physiological parameters. The decline in leaf water potential caused by the chilling exposure to 4°C (day/night) was minimized when GLK-8903 was applied to the plants as compared to the non-treated control. Chilling causes an increase in electrolyte leakage, an indication of chilling injury that occurs at the site of plasma membrane. An increased electrolyte leakage was reduced in the GLK-8903-treated plants during chilling. Data from plasmolysis and deplasmolysis studies of epidermal cells suggest that GLK-8903 is able to stabilize the plasma membrane under stress condition by determining the permeability coefficients plasmometrically (1.96 cm s-1 × 10-4 for GLK-8903-treated plants vs. 4.00 for the controls 3 d at 4°C) with less decreased activity of the plasma membrane ATPase (9.36 μmol ATP.mg chl-1·h-1 for GLK-8903-treated plants vs. 5.04 for the controls 3 d at 4°C). GLK-8903 appears to have high application potential in protecting bean plants from chilling injury with improved yield.

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Hui-lian Xu, Laurent Gauthier, and André Gosselin

Tomato plants (Lycopersicon esculentum Mill. cv. Capello) were grown in peat bags, rockwool slabs, and NFT in a greenhouse to examine the effects of nutrient solution electrical conductivity (EC) and potential evapotranspiration (PET)-dependent EC variation on plant water relations. Peat bags were irrigated by a PET-dependent irrigation system. EC was varied from 1 to 4 mS·cm-1 according to PET under –5 and –9 kPa of substrate water potential setpoints (SWPS). The plants in rockwool and NFT were treated with ECs of 2.5, 4, and 5.5 mS·cm-1. Peat bags and rockwool slabs were overwatered once a week to wash out the accumulated salts. Leaf water potential (ψ1) and relative water content (θ) were measured before and after plants were overwatered. Turgor (P) and osmotic π potentials were estimated from the pressure-volume method. Before plants were overwatered, ψ1 was significantly lower in the plants with high EC and low SWPS treatments and also lower in variable EC-treated plants, but P maintained close to the control value. After plants were overwatered, ψ1 recovered close to the control level and P became higher because of the lower π in the treatments of high EC, variable EC, and/or low SWPS. At a given ψ1 the plants with high EC, variable EC, and/or low SWPS maintained higher θ. The analysis of the pressure-volume curve showed that the leaves treated with high EC, variable EC, and/or low SWPS had higher turgid water content, higher symplasmic (osmotically active) water content, lower apoplasmic (osmotically inactive) water content, and lower θ point of zero turgor (incipient plasmolysis). Maintenance of P after overwatering was directly proportional to photosynthetic capacity. We suggest that osmotic adjustment occurs in response to high EC, low SWPS, or both and that overwatering substrates and varying EC can not only avoid salinity stress, but also improve turgor maintenance.