Carbon metabolism in leaves of transgenic tobacco (Nicotiana tabacum L.) containing elevated fructose 2,6-bisphosphate levels

The aim of this work was to investigate the role of fructose 2,6-bisphosphate (Fru 2,6-P₂) during photosynthesis. The level of Fru 2,6-P₂ in tobacco plants was elevated by the introduction of a modified mammalian gene encoding 6-phosphofructo-2-kinase (6-PF-2-K). Estimates of the metabolite control coefficient (C) for Fru 2,6-P₂ levels in response to increased 6-PF-2-K activity, suggest that small increases in 6-PF-2-K activity have little effect upon steady-state Fru 2,6-P₂ levels (_C = +0.08 for a 0–58% increase in 6-PF-2-K activity). However, larger changes resulted in dramatic rises in Fru 2,6-P₂ levels (C = +3.35 for 206–268% increase in 6-PF-2-K activity). Transgenic plants contained Fru 2,6-P₂ levels in the dark that ranged from 104 to 230% of the level in wild-type tobacco. Plants with altered levels of Fru 2,6-P₂ were used to determine the effects of this signal metabolite upon carbohydrate metabolism during the initial phase of the light period. Here we provide direct evidence that Fru 2,6-P₂ contributes to the regulation of carbon partitioning in tobacco leaves by inhibiting sucrose synthesis.     

The Effect of Elevated Concentrations of Fructose 2,6-Bisphosphate on Carbon Metabolism during Deacidification in the Crassulacean Acid Metabolism Plant Kalanchöe daigremontiana

In C₃ plants, the metabolite fructose 2,6-bisphosphate (Fru 2,6-P₂) has an important role in the regulation of carbon partitioning during photosynthesis. To investigate the impact of Fru 2,6-P₂ on carbon metabolism during Crassulacean acid metabolism (CAM), we have developed an Agrobacterium tumefaciens-mediated transformation system in order to alter genetically the obligate CAM plant Kalanchöe daigremontiana. To our knowledge, this is the first report to use genetic manipulation of a CAM species to increase our understanding of this important form of plant metabolism. Transgenic plants were generated containing a modified rat liver 6-phosphofructo-2-kinase gene. In the plants analyzed the activity of 6-phosphofructo-2-kinase ranged from 175% to 198% of that observed in wild-type plants, resulting in Fru 2,6-P₂ concentrations that were 228% to 350% of wild-type plants after 2 h of illumination. A range of metabolic measurements were made on these transgenic plants to investigate the possible roles of Fru 2,6-P₂ during Suc, starch, and malic acid metabolism across the deacidification period of CAM. The results suggest that Fru 2,6-P₂ plays a major role in regulating partitioning between Suc and starch synthesis during photosynthesis. However, alterations in Fru 2,6-P₂ levels had little effect on malate mobilization during CAM fluxes.     

Banana Ripening: Implications of Changes in Internal Ethylene and CO(2) Concentrations, Pulp Fructose 2,6-Bisphosphate Concentration, and Activity of Some Glycolytic Enzymes

In ripening banana (Musa acuminata L. [AAA group, Cavandish subgroup] cv. Valery) fruit, the steady state concentration of the glycolytic regulator fructose 2,6-bisphosphate (Fru 2,6-P₂) underwent a transient increase 2 to 3 hours before the respiratory rise, but coincident with the increase in ethylene synthesis. Fru 2,6-P₂ concentration subsequently decreased, but increased again approximately one day after initiation of the respiratory climacteric. This second rise in Fru 2,6-P₂ continued as ripening proceeded, reaching approximately five times preclimacteric concentration. Pyrophosphate-dependent phosphofructokinase glycolytic activity exhibited a transitory rise during the early stages of the respiratory climacteric, then declined slightly with further ripening. Cytosolic fructose 1,6-bisphosphatase activity did not change appreciably during ripening. The activity of ATP-dependent phosphofructokinase increased approximately 1.6-fold concurrent with the respiratory rise. A balance in the simultaneous glycolytic and gluconeogenic carbon flow in ripening banana fruit appears to be maintained through changes in substrate levels, relative activities of glycolytic enzymes and steady state levels of Fru 2,6-P₂.     

Stimulation of yeast phosphofructokinase activity by Fructose 2,6-bisphosphate

Fructose 2,6-bisphosphate is a powerful activator of yeast phosphofructokinase when assayed at pH levels of ≥7.0. Half maximal stimulation of enzyme activity occurs at 10^?7 M levels of Fru 2,6-P₂ concentration. This stimulating effect by Fru 2,6-P₂ can be synergistic to that exerted by AMP in counteracting the inhibition of phosphofructokinase activity by ATP. The affinity (S_0.5) of the yeast enzyme to fructose 6-phosphate changes from 1.5 mM in the absence of Fru 2,6-P₂ to 40 μM in its presence.     

Changes in Fructose 2,6-Bisphosphate Levels in Green Pepper (Capsicum annuum L.) Fruit in Response to Temperature

The regulatory metabolite, fructose 2,6-bisphosphate (Fru 2,6-P₂) was found in green pepper (Capsicum annuum L.). The Fru 2,6-P₂ level was found to: (a) rise rapidly in response to heat; (b) drop rapidly, followed by recovery, in response to cold storage of fruit and, (c) oscillate during cold storage of fruit. The possible existence of a relationship between chilling injury and Fru 2,6-P₂ is considered.     

Banana ripening: implications of changes in glycolytic intermediate concentrations, glycolytic and gluconeogenic carbon flux, and fructose 2,6-bisphosphate concentration

In ripening banana (Musa sp. [AAA group, Cavendish subgroup] cv Valery) fruit, the concentration of glycolytic intermediates increased in response to the rapid conversion of starch to sugars and CO₂. Glucose 6-phosphate (G-6-P), fructose 6-phosphate (Fru 6-P), and pyruvate (Pyr) levels changed in synchrony, increasing to a maximum one day past the peak in ethylene synthesis and declining rapidly thereafter. Fructose 1,6-bisphosphate (Fru 1,6-P₂) and phosphoenolpyruvate (PEP) levels underwent changes dissimilar to those of G 6-P, Fru 6-P, and Pyr, indicating that carbon was regulated at the PEP/Pyr and Fru 6-P/Fru 1,6-P₂ interconversion sites. During the climacteric respiratory rise, gluconeogenic carbon flux increased 50- to 100-fold while glycolytic carbon flux increased only 4- to 5-fold. After the climacteric peak in CO₂ production, gluconeogenic carbon flux dropped dramatically while glycolytic carbon flux remained elevated. The steady-state fructose 2,6-bisphosphate (Fru 2,6-P₂) concentration decreased to ½ that of preclimacteric fruit during the period coinciding with the rapid increase in gluconeogenesis. Fru 2,6-P₂ concentration increased thereafter as glycolytic carbon flux increased relative to gluconeogenic carbon flux. It appears likely that the initial increase in respiration in ripening banana fruit is due to the rapid influx of carbon into the cytosol as starch is degraded. As starch reserves are depleted and the levels of intermediates decline, the continued enhancement of respiration may, in part, be maintained by an increased steady-state Fru 2,6-P₂ concentration acting to promote glycolytic carbon flux at the step responsible for the interconversion of Fru 6-P and Fru 1,6-P₂.     

Photosynthetic carbon metabolism in leaves of transgenic tobacco (Nicotiana tabacum L.) containing decreased amounts of fructose 2,6-bisphosphate

The aim of this work was to examine the role of fructose 2,6-bisphosphate (Fru-2,6-P₂) in photosynthetic carbon partitioning. The amount of Fru-2,6-P₂ in leaves of tobacco (Nicotiana tabacum L. cv. Samsun) was reduced by introduction of a modified mammalian gene encoding a functional fructose-2,6-bisphosphatase (EC 3.1.3.46). Expression of this gene in transgenic plants reduced the Fru-2,6-P₂ content of darkened leaves to between 54% and 80% of that in untransformed plants. During the first 30 min of photosynthesis sucrose accumulated more rapidly in the transgenic lines than in the untransformed plants, whereas starch production was slower in the transgenic plants. On illumination, the proportion of ¹⁴CO₂ converted to sucrose was greater in leaf disks of transgenic lines possessing reduced amounts of Fru-2,6-P₂ than in those of the control plants, and there was a corresponding decrease in the proportion of carbon assimilated to starch in the transgenic lines. Furthermore, plants with smaller amounts of Fru-2,6-P₂ had lower rates of net CO₂ assimilation. In illuminated leaves, decreasing the amount of Fru-2,6-P₂ resulted in greater amounts of hexose phosphates, but smaller amounts of 3-phosphoglycerate and dihydroxyacetone phosphate. These differences are interpreted in terms of decreased inhibition of cytosolic fructose-1,6-bisphosphatase resulting from the lowered Fru-2,6-P₂ content. The data provide direct evidence for the importance of Fru-2,6-P₂ in co-ordinating chloroplastic and cytosolic carbohydrate metabolism in leaves in the light. Received: 8 February 2000 / Accepted: 25 April 2000     

Lack of fructose 2,6‐bisphosphate compromises photosynthesis and growth in Arabidopsis in fluctuating environments

The balance between carbon assimilation, storage and utilisation during photosynthesis is dependent on partitioning of photoassimilate between starch and sucrose, and varies in response to changes in the environment. However, the extent to which the capacity to modulate carbon partitioning rapidly through short‐term allosteric regulation may contribute to plant performance is unknown. Here we examine the physiological role of fructose 2,6‐bisphosphate (Fru‐2,6‐P₂) during photosynthesis, growth and reproduction in Arabidopsis thaliana (L.). In leaves this signal metabolite contributes to coordination of carbon assimilation and partitioning during photosynthesis by allosterically modulating the activity of cytosolic fructose‐1,6‐bisphosphatase. Three independent T‐DNA insertional mutant lines deficient in 6‐phosphofructo‐2‐kinase/fructose‐2,6‐bisphosphatase (F2KP), the bifunctional enzyme responsible for both the synthesis and degradation of Fru‐2,6‐P₂, lack Fru‐2,6‐P₂. These plants have normal steady‐state rates of photosynthesis, but exhibit increased partitioning of photoassimilate into sucrose and have delayed photosynthetic induction kinetics. The F2KP‐deficient plants grow normally in constant environments, but show reduced growth and seed yields relative to wildtype plants in fluctuating light and/or temperature. We conclude that Fru‐2,6‐P₂ is required for optimum regulation of photosynthetic carbon metabolism under variable growth conditions. These analyses suggest that the capacity of Fru‐2,6‐P₂ to modulate partitioning of photoassimilate is an important determinant of growth and fitness in natural environments.     

Changes in Activities of Enzymes of Carbon Metabolism in Leaves during Exposure of Plants to Low Temperature

The aim of this study was to determine the response of photosynthetic carbon metabolism in spinach and bean to low temperature. (a) Exposure of warm-grown spinach and bean plants to 10°C for 10 days resulted in increases in the total activities of a number of enzymes, including ribulose 1,5-bisphosphate carboxylase (Rubisco), stromal fructose 1,6 bisphosphatase (Fru 1,6-P₂ase), sedoheptulose 1,7-bisphosphatase (Sed 1,7-P₂ase), and the cytosolic Fru 1,6-P₂ase. In spinach, but not bean, there was an increase in the total activity of sucrose-phosphate synthase. (b) The CO₂-saturated rates of photosynthesis for the cold-acclimated spinach plants were 68% greater at 10°C than those for warm-acclimated plants, whereas in bean, rates of photosynthesis at 10°C were very low after exposure to low temperature. (c) When spinach leaf discs were transferred from 27 to 10°C, the stromal Fru 1,6-P₂ase and NADP-malate dehydrogenase were almost fully activated within 8 minutes, and Rubisco reached 90% of full activation within 15 minutes of transfer. An initial restriction of Calvin cycle fluxes was evident as an increase in the amounts of ribulose 1,5-bisphosphate, glycerate-3-phosphate, Fru 1,6-P₂, and Sed 1,7-P₂. In bean, activation of stromal Fru 1,6-P₂ase was weak, whereas the activation state of Rubisco decreased during the first few minutes after transfer to low temperature. However, NADP-malate dehydrogenase became almost fully activated, showing that no loss of the capacity for reductive activation occurred. (d) Temperature compensation in spinach evidently involves increases in the capacities of a range of enzymes, achieved in the short term by an increase in activation state, whereas long-term acclimation is achieved by an increase in the maximum activities of enzymes. The inability of bean to activate fully certain Calvin cycle enzymes and sucrose-phosphate synthase, or to increase nonphotochemical quenching of chlorophyll fluorescence at 10°C, may be factors contributing to its poor performance at low temperature.     

Fructose 2,6-bisphosphate and the regulation of pyrophosphate-dependent phosphofructokinase activity in germinating pea seeds

The activity of pyrophosphate: d-fructose-6-phosphate-1-phosphotransferase (EC 2.7.1.90, PPi-PFK) in cotyledons and sprouts of germinating pea seeds (Pisum sativum cv Alaska or Green Arrow) increases rapidly during the first 2 to 3 days after imbibition and then declines to a lower activity. The reaction toward fructose 1,6-bisphosphate formation is activated greatly by fructose 2,6-bisphosphate (fru 2,6-P₂); however, the sensitivity of the enzyme's activity to fru 2,6-P₂ activation changes during germination.     

Is xanthine dehydrogenase involved in response of pea plants (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.) to salinity or ammonium treatment?

The effect of salinity and different nitrogen sources on the level of xanthine dehydrogenase (XDH) activity in roots and leaves of pea plants was investigated. Two bands of xanthine dehydrogenase activity (XDH-R2, XDH-R3) were detected in roots after native PAGE and staining with hypoxanthine as substrate. Only one band of XDH activity (XDH-L1) was detected in leaf extracts. Within leaves of three different ages the highest XDH activity was detected in young leaves both under control as well as stress conditions. Salinity did not affect significantly the activity of XDH in pea roots, however, depressed XDH activity in leaves. A significant increase of XDH activity both in roots and leaves was observed only when ammonium was applied as the sole N source. Increased concentration of ureides in the xylem sap of pea plants was observed for both ammonium and high salt treatments, although the higher content of ureides in the xylem sap of 100 mM NaCl treated plants may be rather a result of lower rate of exudation from roots than of increased root ureide biosynthesis. Thus, the changes of root and leaf XDH activity in pea plants seem to be tightly correlated with ureide synthesis that is induced by NH ₄ ⁺ , the product of N fixation, and rather than by salinity. A contribution of pea XDH in increased oxygen species or uric acid production under saline conditions seems to be less than likely.     

In vitro activation of phosphoglucomutase by fructose 2,6-bisphosphate

The hexose bisphosphate activation of phosphoglucomutase was investigated with both plant (pea and mung bean) and animal (rabbit muscle) sources of the enzyme. Plant phosphoglucomutase was purified about 50-fold from seeds, and to a lesser extent, from seedlings of Pisum sativum L. cv Grenadier and seedlings of Phaseolus aureus. It was found that the plant enzyme was isolated in a mostly dephosphorylated form while commercial rabbit muscle phosphoglucomutase was predominantly in the phosphorylated form. Activation studies were done using the dephosphorylated enzymes. The range of activation constant (K_a) values were obtained for each bisphosphate were: for glucose 1-6-P₂, 0.5 to 1.8; fructose 2,6-P₂, 6 to 11.7; and fructose 1,6-P₂, 7 micromolar, respectively. Fructose 2,6-P₂ is known to occur in both plant and animal tissues at changing levels encompassing the K_a values found in this study; hence, these results implicate fructose 2,6-P₂ as a natural activator of phosphoglucomutase, particularly in plants. Also, glucose 1,6-P₂ has not been found in plants, and the method for measuring glucose 1,6-P₂ by monitoring the activation of phosphoglucomutase is not specific.     

涝渍对3个树种生长、组织孔隙度和渗漏氧的影响

为了了解落羽杉(Taxodium distichum)、乌桕(Sapium sebiferum)和美国山核桃(Carya illinoensis)等树种的耐涝机制, 采用盆栽模拟涝渍环境的试验方法, 设置了淹水、渍水和对照3个处理, 测定了一年生落羽杉、乌桕和美国山核桃实生苗的生长、组织孔隙度、根氧消耗等指标。结果表明, 涝渍处理抑制了落羽杉、乌桕和美国山核桃的生物量和生物量增量(渍水处理下落羽杉的生长得到了促进), 增加了3树种的根冠比, 从生物量和生物量增量下降幅度来评价, 落羽杉的耐涝性最强, 其次为美国山核桃。淹水和渍水处理下, 落羽杉、乌桕和美国山核桃的根、茎和叶中的组织孔隙度显著增加, 且随着处理时间的延长, 各器官的组织孔隙度有增加的趋势, 3个树种中, 落羽杉的根、茎和叶中的组织孔隙度均较其他2个树种高。淹水和渍水处理下, 移除茎明显增加了落羽杉、美国山核桃和乌桕的根的氧消耗, 表明涝渍处理增强了O₂在3个树种体内的运输并通过根系扩散到涝渍土壤中的能力, 并且随着处理时间的延长, 3个树种体内运输O₂并扩散到土壤中的能力有逐渐增强的趋势。因此, 涝渍环境总体上抑制了落羽杉、乌桕和美国山核桃等树种的生长, 但各树种为了适应这种生长环境, 形成了大量的通气组织, 从而导致各器官组织孔隙度的增加, 增强了O₂通过植物体运输到根系并扩散到土壤中的能力, 解决了根系及根际缺氧的矛盾。     

A role for fructose 2,6-bisphosphate in regulating carbohydrate metabolism in guard cells

Fructose 2,6-bisphosphate (Fru2,6P₂) appears to function as a regulator metabolite in glycolysis and gluconeogenesis in animal tissues, yeast, and the photosynthetic cells of leaves. We have investigated the role of Fru2,6P₂ in guard-cell protoplasts from Vicia faba L. and Pisum sativum L. (Argenteum mutant), and in epidermal strips purified by sonication from all cells except for the guard cells. Guard-cell protoplasts were separated into fractions enriched in cytosol and in chloroplasts by passing them through a nylon net, followed by silicone oil centrifugation. The cytosol contained a pyrophosphate: fructose 6-phosphate phosphotransferase (involved in glycolysis) which was strongly stimulated by Fru2,6P₂. A cytosolic fructose 1,6-bisphosphatase (a catalyst of gluconeogenesis) was inhibited by Fru2,6P₂. There was virtually no fructose 1,6-bisphosphatase activity in guard-cell chloroplasts of V. faba. It is therefore unlikely that the starch formed in these chloroplasts originates from imported triose phosphates or phosphoglycerate.

The level of Fru2,6P₂ in guard-cell protoplasts and epidermal strips was about 0.1 to 1 attomole per guard cell in the dark (corresponding to 0.05 to 0.5 nanomole per milligram chlorophyll) and increased three- to tenfold within 15 minutes in the light. Within the same time span, hexose phosphate levels in guard-cell protoplasts declined to approximately one-half, indicating that acceleration of glycolysis involved stimulation of reactions using hexose phosphates. The level of Fru2,6P₂ in guard cells appears to determine the direction in which carbohydrate metabolism proceeds.

     

Regulation of Carbon Partitioning in Source and Sink Leaf Parts in Sweet Pepper (Capsicum annuum L.) Plants : Role of Fructose 2,6-Bisphosphate

Area expansion rate, partitioning of photosynthetically fixed carbon, and levels of fructose 2,6-bisphosphate (fru-2,6-P₂) were determined in individual parts of developing leaves of sweet pepper (Capsicum annuum L.). The base was rapidly expanding and allocated less carbon to sucrose synthesis in comparison to the leaf tip, where expansion had almost stopped. The change in leaf expansion rate and carbon partitioning happened gradually. During day time levels of fru-2,6-P₂ were consistently higher in the leaf base than in the leaf tip. Leaf expansion rate and carbon partitioning were closely related to day time levels of fru-2,6-P₂, suggesting that fru-2,6-P₂ is an important factor in adjustment of metabolism during sink-to-source transition of leaf tissue. The levels of fru-2,6-P₂ changed markedly after a dark-to-light transition in the leaf base, but not in the leaf tip, suggesting that regulatory systems based on fru-2,6-P₂ are different in sink and source leaf tissue. During the period upon dark-to-light transition the variations in level of fru-2,6-P₂ did not show a close correlation to changes in the carbon partitioning, until the metabolism had reached a steady state.     

Rapid modulation of nitrate reductase in pea roots

The regulatory properties of nitrate reductase (NR; EC 1.6.6.1) in root extracts from hydroponically grown pea (Pisum sativum L. cv. Kleine Rheinländerin) plants were examined and compared with known properties of NR from spinach and pea leaves. Nitrate-reductase activity (NRA) extracted from pea roots decreased slowly when plants were kept in the dark, or when illuminated plants were detopped, with a half-time of about 4 h (= slow modulation in vivo). In contrast, the half-time for the dark-inactivation of NR from pea leaves was only 10 min. However, when root tip segments were transferred from aerobic to anaerobic conditions or vice versa, changes in NRA were as rapid as in leaves (= rapid modulation in vivo). Nitrate-reductase activity was low when extracted from roots kept in solutions flushed with air or pure oxygen, and high in nitrogen. Okadaic acid, a specific inhibitor of type-1 and type-2A protein phosphatases, totally prevented the in vivo activation by anaerobiosis of NR, indicating that rapid activation of root NR involved protein dephosphorylation. Under aerobic conditions, the low NRA in roots was also rapidly increased by incubating the roots with either uncouplers or mannose. Under these conditions, and also under anaerobiosis, ATP levels in roots were much lower than in aerated control roots. Thus, whenever ATP levels in roots were artificially decreased, NRA increased rapidly. The highly active NR extracted from anaerobic roots could be partially inactivated in vitro by preincubation of desalted root extracts with MgATP (2 mM), with a half-time of about 20 min. It was reactivated by subsequently incubating the extracts with excess AMP (2 mM). Thus, pea root NR shares many of the previously described properties of NR from spinach leaves, suggesting that the root enzyme, like the leaf enzyme, can be rapidly modulated, probably by reversible protein phosphorylation/ dephosphorylation.     

Can plants exposed to SO2 excrete sulfuric acid through the roots?

Hydroponically grown pea plants (Pisum sativum L., cv. Kleine Rheinländerin) and barley seedlings (Hordeum vulgare L., cv. Gerbel) were fumigated for several days with 1 or 2 μl l^?1 SO₂. Both species accumulated sulfate during fumigation, although the nutrient medium lacked sulfate. In pea, SO₂-dependent sulfate accumulation in different plant parts accounted for 60 percent of the SO₂ sulfur which, as calculated from a determination of boundary and stomatal flux resistances had entered the leaves. Up to 55% of the air-borne sulfate was translocated from pea leaves to roots during the period of fumigation, but no or only little sulfate was excreted into the nutrient solution. In contrast, barley retained sulfate in the leaves, and sulfate translocation from shoot to the root system could not be observed. In both species, protons were excreted by the roots. In fumigated plants, proton loss was higher than in untreated controls in pea, but not in barley. In pea, SO₂-dependent proton loss into the medium accounted for up to 50% of the sulfuric acid formed from SO₂. Proton excretion was strongly dependent on potassium availability in the nutrient medium. Cation uptake by the plants during fumigation was sufficient to compensate for proton loss, suggesting proton/cation exchange at the interface between root and medium. We conclude that by oxidation to sulfuric acid, plants are capable of detoxifying SO₂ taken up by the leaves. Depending on plant species, either both protons and sulfate anions can be exported from the leaves, or the proton load on leaf cells can be relieved by proton/cation exchange at the plasmalemma. Finally, the problem of airborne plant acidification may be solved by proton/cation exchange at the level of roots. The burden of acidification is then shifted from the plant to the nutrient medium. Appreciable amounts of sulfate can be excreted neither by pea nor by barley plants.     

ABA in Roots and Leaves of Flooded Pea Plants

Zhang, J. and Davies, W. J. 1987. ABA in roots and leaves offlooded pea plants.—J. exp. Bot. 38: 649–659. Roots of potted pea (Pisum sativum L. cv. Feltham First) seedlingswere flooded with tap water. Within a few hours of the startof the flooding treatment the content of free ABA in roots increasedcompared to contents of roots of unflooded control plants butthis increase was not statistically significant until the beginningof the second day after flooding. Approximately 36 h after firstflooding significant increases in the free-ABA content of leaveswere detected. This was 14 h after significant increases inthe amount of ABA in the roots of the same plants. There wasmarked diurnal variation in free-ABA content of leaves and rootsof plants that had been flooded for several days, with maximumcontents recorded 3 h or more after the beginning of the lightperiod. Very rapidly after the lights were switched oft ABAcontents declined. On day 3 of the flooding treatment, therewas more than a 5-fold decrease in the free-ABA content of leaveswithin a few hours of the beginning of the dark period. Radio-immunoassaysuggested that a very large proportion of the total ABA in theplant was in a bound form. This form of ABA increased substantiallyas the flooding period progressed. The importance of variation in ABA content for the control ofwater relations and gas exchange of flooded plants is discussed. Key words: Flooding, Pisum sativum, ABA, water relations     

Positive and Negative Messages from Roots Induce Foliar Desiccation and Stomatal Closure in Flooded Pea Plants

Flooding the soil for 5–7 d caused partial desiccationin leaves of pea plants (Pisum sativum. L. cv. ‘Sprite’).The injury was associated with anaerobiosis in the soil, a largeincrease in the permeability of leaf tissue to electrolytesand other substances, a low leaf water content and an increasedwater saturation deficit (WSD). Desiccating leaves also lackedthe capacity to rehydrate in humid atmospheres, a disabilityexpressed as a water resaturation deficit (WRSD). This irreversibleinjury was preceded during the first 4–5 d of floodingby closure of stomata within 24 h, decreased transpiration,an unusually large leaf water content and small WSD. Leaf waterpotentials were higher than those in well-drained controls.Also, there was no appreciable WRSD. Leaflets detached fromflooded plants during this early phase retained their watermore effectively than those from controls when left exposedto the atmosphere for 5 min. Stomatal closure and the associated increase in leaf hydrationcould be simulated by excising leaves and incubating them withtheir petioles in open vials of water. Thus, such changes inflooded plants possibly represented a response to a deficiencyin the supply of substances that would usually be transportedfrom roots to leaves in healthy plants (negative message). Ionleakage and the associated loss of leaf hydration that occurswhen flooding is extended for more than 5 d could not be simulatedby isolating the leaves from the roots. Appearance of this symptomdepended on leaves remaining attached to flooded root systems,implying that the damage is caused by injurious substances passingupwards (positive message). Both ethylene and ethanol have beeneliminated as likely causes, but flooding increased phosphorusin the leaves to concentrations that may be toxic. Key words: Pisum sativum, Flooding, Foliar desiccation, Stomata, Ethylene