Effect of drought stress on net CO2 uptake by Zea leaves

The net CO₂ assimilation by leaves of maize (Zea mays L. cv. Adonis) plants subjected to slow or rapid dehydration decreased without changes in the total extractable activities of phosphoenolpyruvate carboxylase (PEPC), malate dehydrogenase (MDH) and malic enzyme (ME). The phosphorylation state of PEPC extracted from leaves after 2–3 h of exposure to light was not affected by water deficit, either. Moreover, when plants which had been slowly dehydrated to a leaf relative water content of about 60% were rehydrated, the net CO₂ assimilation by leaves increased very rapidly without any changes in the activities of MDH, ME and PEPC or phosphorylation state of PEPC. The net CO₂-dependent O₂ evolution of a non-wilted leaf measured with an oxygen electrode decreased as CO₂ concentration increased and was totally inhibited when the CO₂ concentration was about 10%. Nevertheless, high CO₂ concentrations (5–10%) counteracted most of the inhibitory effect of water deficit that developed during a slow dehydration but only counteracted a little of the inhibitory effect that developed during a rapid dehydration. In contrast to what could be observed during a rapidly developing water deficit, inhibition of leaf photosynthesis by cis-abscisic acid could be alleviated by high CO₂ concentrations. These results indicate that the inhibition of leaf net CO₂ uptake brought about by water deficit is mainly due to stomatal closure when a maize plant is dehydrated slowly while it is mainly due to inhibition of non-stomatal processes when a plant is rapidly dehydrated. The photosynthetic apparatus of maize leaves appears to be as resistant to drought as that of C₃ plants. The non-stomatal inhibition observed in rapidly dehydrated leaves might be the result of either a down-regulation of the photosynthetic enzymes by changes in metabolite pool sizes or restricted plasmodesmatal transport between mesophyll and bundle-sheath cells.     

Synthesis and movement of abscisic Acid in water-stressed cotton leaves

Synthesis and movement of abscisic acid (ABA) into the apoplast of water-stressed cotton (Gossypium hirsutum L.) leaves were examined using pressure dehydration techniques. The exudates of leaves dehydrated in a pressure chamber contained ABA. The level of ABA in the exudates was insensitive to the leaf water potential when dehydration occurred over a 3-hour period. When leaves were rapidly dehydrated in the pressure chamber and held at a balance pressure coincident with the point of zero turgor, ABA accumulated in the leaf tissue and then in the apoplast, but only after 2 to 3 hours of zero turgor. Slow dehydration of leaves by equilibration over varying mannitol concentrations resulted in some accumulation of ABA prior to the point of zero turgor, but ABA accumulated in the tissue and apoplast most rapidly after the onset of zero turgor.     

Effect of cold hardening on the phenolic complex of winter wheat leaves

The effect of hardening on the composition of phenolic compounds in winter wheat (Triticum aestivum L.) leaves was studied. It was shown that green tissues contained mainly flavonoids, especially flavons (C-and O-glycosides of apigenin and luteolin), and also ferulic acid derivatives. Among flavons, derivatives of luteolin dominated, including isoorientin, which comprised approximately a half of the content of all identified phenolic compounds. Low temperature induced the accumulation of phenolic compounds in winter wheat leaves, whereas their qualitative composition was not practically changed.     

Rapid Osmotic Adjustment in Detached Wheat Leaves

Osmotic adjustment is induced in detached wheat leaves by rapiddrying to a relative water content below 0·65, followedby re-saturation. Quantitatively, the response to this treatmentis comparable to the maximum of adjustment obtained with pottedplants at the same developmental stage. Low temperatures duringdrying and re-saturation of the leaves reduce the adjustmentresponse. We conclude that drought stress serves as a triggeronly, while the metabolic events lowering the osmotic potentialare favoured by high or intermediate water contents. Triticum durum L., durum wheat, rapid dehydration, osmotic adjustment, pressure-volume curves     

Effect of microtubule stabilization on the freezing tolerance of mesophyll cells of spinach

Freezing, dehydration, and supercooling cause microtubules in mesophyll cells of spinach (Spinacia oleracea L. cv Bloomsdale) to depolymerize (ME Bartolo, JV Carter, Plant Physiol [1991] 97: 175-181). The objective of this study was to determine whether the LT₅₀ (lethal temperature: the freezing temperature at which 50% of the tissue is killed) of spinach leaf tissue can be changed by diminishing the extent of microtubule depolymerization in response to freezing. Also examined was how tolerance to the components of extracellular freezing, low temperature and dehydration, is affected by microtubule stabilization. Leaf sections of nonacclimated and cold-acclimated spinach were treated with 20 micromolar taxol, a microtubule-stabilizing compound, prior to freezing, supercooling, or dehydration. Taxol stabilized microtubules against depolymerization in cells subjected to these stresses. When pretreated with taxol both nonacclimated and cold-acclimated cells exhibited increased injury during freezing and dehydration. In contrast, supercooling did not injure cells with taxol-stabilized microtubules. Electrolyte leakage, visual appearance of the cells, or a microtubule repolymerization assay were used to assess injury. As leaves were cold-acclimated beyond the normal period of 2 weeks taxol had less of an effect on cell survival during freezing. In leaves acclimated for up to 2 weeks, stabilizing microtubules with taxol resulted in death at a higher freezing temperature. At certain stages of cold acclimation, it appears that if microtubule depolymerization does not occur during a freeze-thaw cycle the plant cell will be killed at a higher temperature than if microtubule depolymerization proceeds normally. An alternative explanation of these results is that taxol may generate abnormal microtubules, and connections between microtubules and the plasma membrane, such that normal cellular responses to freeze-induced dehydration and subsequent rehydration are blocked, with resultant enhanced freezing injury.     

Changes in Chlorophyll Fluorescence in Maize Plants with Imposed Rapid Dehydration at Different Leaf Ages

A comparison of the effects of a rapidly imposed water deficit with different leaf ages on chlorophyll a fluorescence and gas exchange was performed in maize (Zea mays L.) plants. The relationships between photosynthesis and leaf relative turgidity (RT) and ion leakage were further investigated. Leaf dehydration substantially decreased net photosynthetic rate (A) and stomatal conductance (G _s), particularly for older leaves. With dehydration time, F _v /F _m maintained a relatively stable level for youngest leaves but significantly decreased for the older leaves. The electron transport rate (ETR) sharply decreased with intensifying dehydration and remained at lower levels during continuous dehydration. The photochemical quenching of variable chlorophyll fluorescence (q _P) gradually decreased with dehydration intensity for the older leaves but increased for the youngest leaves, whereas dehydration did not affect the nonphotochemical chlorophyll fluorescence quenching (NPQ) for the youngest leaves but remarkably decreased it for the older leaves. The leaf RT was significantly and positively correlated with its F _v /F _m, ETR, and q _P, and the leaf ion leakage was significantly and negatively correlated with F _v /F _m and NPQ. Our results suggest that the photosynthetic systems of young and old leaves decline at different rates when exposed to rapid dehydration.     

Effects of high-temperature stress on various biomembranes of leaf cells in situ and in vitro

The sensitivity of photosynthetic and respiratory functions to supraoptimal temperature stress was compared after heating of leaves, protoplasts and membrane systems of spinach (Spinacia oleracea L. cv. Monatol) and lettuce (Valerianella locusta [L.] Betcke) in situ and in vitro.

After heating of whole leaves or protoplasts, endogenous respiration was not or only slightly affected at temperatures which caused a marked decrease of photosynthesis. This was manifested when mitochondria and thylakoids were isolated from heat-treated leaves. In the presence of exogenous substrates, mitochondrial electron transport and phosphorylation were even somewhat stimulated compared to the controls.

Inactivation of net CO₂ uptake of whole leaves following heat stress and of the photochemical activities of chloroplast membranes isolated from heat-treated leaves of the same origin occurred nearly simultaneously. In protoplasts, photosynthesis was inactivated at temperatures far below those which caused drastic changes in the integrity of the tonoplast and the plasmalemma. This indicates that damage occurring within the chloroplasts rather than alterations in the compartmentation of the cell is responsible for the high sensitivity of photosynthesis to supraoptimal temperature stress.

Mitochondria and thykaloids isolated from the same preparation of intact leaves under comparable conditions and subjected to heat treatment in vitro, however, were inactivated nearly in the same temperature range. Thus, mitochondria are much more stable within their cytoplasmic environment.

     

Freezing tolerance of citrus, spinach, and petunia leaf tissue : osmotic adjustment and sensitivity to freeze induced cellular dehydration

Seasonal variations in freezing tolerance, water content, water and osmotic potential, and levels of soluble sugars of leaves of field-grown Valencia orange (Citrus sinensis) trees were studied to determine the ability of citrus trees to cold acclimate under natural conditions. Controlled environmental studies of young potted citrus trees, spinach (Spinacia pleracea), and petunia (Petunia hybrids) were carried out to study the water relations during cold acclimation under less variable conditions. During the coolest weeks of the winter, leaf water content and osmotic potential of field-grown trees decreased about 20 to 25%, while soluble sugars increased by 100%. At the same time, freezing tolerance increased from lethal temperature for 50% (LT₅₀) of −2.8 to −3.8°C. In contrast, citrus leaves cold acclimated at a constant 10°C in growth chambers were freezing tolerant to about −6°C. The calculated freezing induced cellular dehydration at the LT₅₀ remained relatively constant for field-grown leaves throughout the year, but increased for leaves of plants cold acclimated at 10°C in a controlled environment. Spinach leaves cold acclimated at 5°C tolerated increased cellular dehydration compared to nonacclimated leaves. Cold acclimated petunia leaves increased in freezing tolerance by decreasing osmotic potential, but had no capacity to change cellular dehydration sensitivity. The result suggest that two cold acclimation mechanisms are involved in both citrus and spinach leaves and only one in petunia leaves. The common mechanism in all three species tested was a minor increase in tolerance (about −1°C) resulting from low temperature induced osmotic adjustment, and the second in citrus and spinach was a noncolligative mechanism that increased the cellular resistance to freeze hydration.     

A water potential threshold for the increase of abscisic Acid in leaves

A relationship between abscisic acid concentration and leaf water status is reported. Water potentials were measured in leaves of Ambrosia artemisiifolia L. and Ambrosia trifida L. throughout a period of dehydration of intact plants. Tissues from the same leaves were analyzed for abscisic acid. For both species, abscisic acid began to increase in a critical water potential range (−10 to −12 atmospheres). These data suggest a threshold water potential that stimulates abscisic acid synthesis. The data support the hypothesis that a small change in water potential could affect stomatal resistance to water loss by means of a very sensitive chemical feedback control mechanism.     

Occurrence of a high temperature sensitivity of chloroplast ribosome formation in several higher plants

A specific high temperature-induced deficiency of chloroplast ribosome formation, as indicated by the absence of chloroplast rRNA, has been observed in the leaves of light- or dark-grown seedlings of Avena sativa L., Hordeum vulgare L., and Triticum aestivum L. at certain temperatures between 28 and 34 C. While the growth of the leaves (size, morphology, total amino nitrogen content) was little affected by the elevated temperature, chlorophyll accumulation was strongly inhibited, amounting to only 2 to 20% of its content in 22 C-grown leaves which were used as a reference for normal development. The carotenoid contents were also lower but still reached at least 15 to 20% of the corresponding measurements at 22 C. Ribulose-1,5-bisphosphate carboxylase was absent at the higher temperature while NADP-glyceralde-hydephosphate dehydrogenase reached high activities. For the peroxisomal marker enzyme hydroxypyruvate reductase, 30 to 70% of the activity present in 22 C-grown leaves was found in extracts from high temperature-grown leaves. Fumarase reached 1.5- to 4-fold higher activities at the elevated growth temperature than at 22 C. Leaves of Pisum sativum L. were completely chlorotic and deficient of 70S ribosomes at 33 C but simultaneously suffered from a severe general inhibition of their growth. In Zea mays L., a formation of chlorotic leaves was not observed at elevated temperatures.     

Water Relations of Cotton Plants under Nitrogen Deficiency: V. Environmental Control of Abscisic Acid Accumulation and Stomatal Sensitivity to Abscisic Acid

Suboptimal N nutrition increased the water potential for stomatal closure in water stressed cotton (Gossypium hirsutum L.) leaves. This increased sensitivity to water stress had two components, increased accumulation of abscisic acid (ABA) and increased apparent stomatal sensitivity to ABA. Low N increased the threshold water potentials for stomatal closure and ABA accumulation by about 4 bars and 2 bars, respectively. Low N also greatly increased stomatal response to low concentrations of exogenous ABA applied to excised leaves through the transpiration stream. In low N leaves, kinetin decreased stomatal response to ABA to the level observed with high N leaves. Kinetin by itself had little effect on stomata, nor did it alter stomatal response to ABA in high N leaves. The results suggest a cytokinin-ABA balance which is altered by suboptimal N nutrition to favor stomatal closure during stress.

Ambient temperature and N nutrition interacted to alter stomatal response to water stress. Stress-induced ABA accumulation and apparent stomatal sensitivity to ABA were independently affected. The effects of each treatment, and their interaction, could be explained as the net result of changes in both accumulation and apparent sensitivity. Although the results document environmental control of stomatal response to ABA, either altered partitioning of ABA between active and inactive pools, or altered sensitivity of the guard cells, could account for the data.

     

The role of abscisic Acid in cross-adaptation of tobacco plants

Tobacco plants (Nicotiana rustica L.) pre-exposed to leaf dehydration, mineral deprivation, salination, or BO₃³⁻ toxicity exhibited increased resistance to subzero temperature and to reduced oxygen in the root medium. The stressed plants all showed an elevated content of leaf abscisic acid. Upon transfer of mineral deprived and salinated plants to prestress conditions, a decline in leaf abscisic acid content to prestress levels took place together with a loss of the increased resistance to subzero temperature and to deprivation of root oxygen. Treatment with abscisic acid by direct application to the leaves or by addition to the root medium improved leaf resistance to subzero temperature and to deprivation of root oxygen. A common hormone-regulation mechanism involving abscisic acid is suggested for this phenomenon of “cross-adaptation” by which a given stress confers increased resistance to other, apparently unrelated stresses.     

Effects of chilling on the biochemical and functional properties of thylakoid membranes

The mechanism of chilling resistance was investigated in 4-week-old plants of the chilling-sensitive cultivated tomato, Lycopersicon esculentum Mill. cv H722, and rooted cuttings of its chilling-resistant wild relative, L. hirsutum Humb. and Bonpl., which were chilled for 3 days at 2°C with a 14-hour photoperiod and light intensity of 250 micromoles per square meter per second. This chilling stress reduced the chlorophyll fluorescence ratio, stomatal conductance, and dry matter accumulation more in the sensitive L. esculentum than in the resistant L. hirsutum. Photosynthetic CO₂ uptake at the end of the chilling treatment was reduced more in the resistant L. hirsutum than in L. esculentum, but recovered at a faster rate when the plants were returned to 25°C. The reduction of the spin trap, Tiron, by isolated thylakoids at 750 micromoles per square meter per second light intensity was taken as a relative indication of the tendency for the thylakoids to produce activated oxygen. Thylakoids isolated from the resistant L. hirsutum with or without chilling treatment were essentially similar, whereas those from chilled leaves of L. esculentum reduced more Tiron than the nonchilled controls. Whole chain photosynthetic electron transport was measured on thylakoids isolated from chilled and control leaves of the two species at a range of assay temperatures from 5 to 25°C. In both species, electron transport of the thylakoids from chilled leaves was lower than the controls when measured at 25°C, and electron transport declined as the assay temperature was reduced. However, the temperature sensitivity of thylakoids from chilled L. esculentum was altered such that at all temperatures below 20°C, the rate of electron transport exceeded the control values. In contrast, the thylakoids from chilled L. hirsutum maintained their temperature sensitivity, and the electron transport rates were proportionately reduced at all temperatures. This sublethal chilling stress caused no significant changes in thylakoid galactolipid, phospholipid, or protein levels in either species. Nonchilled thylakoid membranes from L. hirsutum had fourfold higher levels of the fatty acid 16:1, than those from L. esculentum. Chilling caused retailoring of the acyl chains in L. hirsutum but not in L. esculentum. The chilling resistance of L. hirsutum may be related to an ability to reduce the potential for free radical production by close regulation of electron transport within the chloroplast.     

Effect of low and high temperatures on the photosynthetic performance of Lantana camara L. Leaves in darkness

Low and high temperatures are known as most important factors influencing plant performance and distribution. Plants of Lantana camara L. coming from two distinct geographical populations (Iberian Peninsula and Galápagos Islands) were cultivated in a common garden experiment, and their leaves were subjected to thermal treatments (from +20.0 to ?7.5°C during the winter and from +20.0 to +50.0°C during the summer) in a programmable water bath in darkness. Their photosynthetic performance and their recovery capacity after the thermal treatment were evaluated by measuring chlorophyll fluorescence, net photosynthesis rate, and leaf necrosis. In general, L. camara photosynthetic apparatus showed a wide range of temperature tolerance in darkness, showing optimal functioning of its photosystem II just after exposure to temperatures between ?2.5 and +35.0°C for the Iberian population and between +10.0 and +25.0°C for the Galápagos population. Just after exposure to low and high temperatures, gradual cold and heat-induced photoinhibition was recorded for both populations. After 24 h, leaves of L. camara demonstrated a great recovery capacity from ?2.5 to +42.5°C. However, leaves of the treatments from ?5.0°C down and +47.50°C up showed permanent damages to the photosynthetic apparatus and to the leaf tissues. Slight interpopulation differences were found only at extreme temperatures.     

Inhibition of Abscisic Acid 8′-Hydroxylase Affects Dehydration Tolerance and Root Formation in Cuttings of Grapes (Vitis labrusca L. × Vitis vinifera L. cv. Kyoho) Under Drought Stress Conditions

The effects of an inhibitor (Abz-E3M) of abscisic acid (ABA) 8′-hydroxylase, which is a primary enzyme of ABA catabolism, on dehydration tolerance and root formation in grape cuttings under drought conditions were investigated. Cuttings of ‘Kyoho’ grape (Vitis labrusca L. × Vitis vinifera L.) were sprayed with 100 μM of Abz-E3M and subjected to water deficit conditions at the stage when their first leaves fully expanded. The physiological and morphological changes in the leaves and basal portions of the cuttings were determined. In Abz-E3M-treated leaves, lower ABA metabolite and higher ABA and indole-3-acetic acid (IAA) concentrations were observed. Compared to the untreated control leaves, higher water potential was significantly maintained in Abz-E3M-treated leaves. Abz-E3M applications resulted in lower proline accumulation and 2,2-diphenyl-2-picrylhydrazyl radical scavenging activity in the leaves and led to enhanced dehydration tolerance. In addition, the percentage of rooted cuttings was significantly increased by Abz-E3M application. In the basal portion of Abz-E3M-treated cuttings, endogenous IAA concentrations and the gene expressions of VvARF6 and VvARF8, which are positive regulators of adventitious root formation, were significantly increased. Moreover, the expression levels of the negative regulator, VvARF17, were significantly lower. These results suggested that the inhibition of ABA 8′-hydroxylase enhanced dehydration tolerance and adventitious rooting and may be an effective strategy for achieving drought stress tolerance in grape cuttings.

     

The relation of carbon dioxide compensation and chlorenchymatous vascular bundle sheaths in leaves of dicots

Low CO₂ compensation points have been found to be associated with several unusual characteristics related to photosynthesis. One such characteristic is a prominent, chlorenchymatous vascular bundle sheath in the leaves. It has been suggested that the presence of this sheath in dicotyledons can serve as a means of detecting low CO₂-compensating species. We collected 88 dicotyledon species from 22 families reported to have chlorenchymatous sheaths. Of the 88, only three, Tribulus terrestris, L., Boerhaavia paniculata, L. C. Rich, and Trianthema portulacastrum L., had low CO₂ compensation points. Cross sections of the leaves of the other species revealed that they did have chlorenchymatous vascular bundle sheaths. However, these sheath cells contained chloroplasts which were not specialized for starch formation as were the bundle sheath chloroplasts of the low CO₂-compensating species.     

Expression of "Dehydrin-Like" Proteins in Embryos and Seedlings of Zizania palustris and Oryza sativa during Dehydration

Proteins inducible by dehydration and abscisic acid (ABA), termed dehydrins or RAB (Responsive to ABA) proteins, have been identified in a number of species and have been suggested to play a role in desiccation tolerance, particularly during seed development. Seeds (caryopses) of North American wild rice (Zizania palustris var interior [Fassett] Dore) are tolerant of dehydration to <10% moisture content (fresh weight basis) only under restricted dehydration and rehydration conditions. In comparison, seeds of paddy rice (Oryza sativa L.) readily tolerate desiccation to <5% water content. Expression of “dehydrin-like” proteins in Zizania and Oryza seedlings and embryos was examined to investigate the relationship between the presence of such proteins and desiccation tolerance. [³⁵S]Methionine labeling of newly synthesized proteins showed that seedlings (first leaf stage) of both Zizania and Oryza synthesized a novel “heat-stable” protein of apparent molecular weight = 20,000 when dehydrated to <75% of their initial fresh weight. ABA (100 micromolar) induced synthesis of a protein with similar electrophoretic mobility in both species. Western blots using antiserum raised against maize (Zea mays L.) dehydrin detected a protein band from dehydrated Zizania shoots and mature embryonic axes that comigrated with the labeled 20-kilodalton polypeptide. Northern blots using a cDNA for an ABA-responsive protein from Oryza (rab 16a) showed that both seedlings and excised embryonic axes of Zizania accumulated RNA similar in sequence to rab 16a in response to water loss. Zizania seedlings and embryonic axes were also capable of ABA accumulation during dehydration. The intolerance of Zizania seeds to dehydration at low temperature is apparently not due to an absence of dehydrin-like proteins or an inability to accumulate ABA.     

Action de l’acide indolyl-3-Acétique sur la déshydratation des limbes chez quatre espèces de blé avec différent niveau de ploïdie

It was found that the greatest dehydration rate of detached leaves indicates theTriticum monococcum L. v.vulgare (2n = 14). The indol-3-Acetic acid distinctly decreases the dehydration rate of detached leaves of examined wheat with a different ploidy level (Triticum monococcum, Triticum dicoccum, Triticum spelta andTriticum aestivum).