Drought, abscisic acid and transpiration rate effects on the regulation of PIP aquaporin gene expression and abundance in Phaseolus vulgaris plants

BACKGROUND AND AIMS: Drought causes a decline of root hydraulic conductance, which aside from embolisms, is governed ultimately by aquaporins. Multiple factors probably regulate aquaporin expression, abundance and activity in leaf and root tissues during drought; among these are the leaf transpiration rate, leaf water status, abscisic acid (ABA) and soil water content. Here a study is made of how these factors could influence the response of aquaporin to drought. METHODS: Three plasma membrane intrinsic proteins (PIPs) or aquaporins were cloned from Phaseolus vulgaris plants and their expression was analysed after 4 d of water deprivation and also 1 d after re-watering. The effects of ABA and of methotrexate (MTX), an inhibitor of stomatal opening, on gene expression and protein abundance were also analysed. Protein abundance was examined using antibodies against PIP1 and PIP2 aquaporins. At the same time, root hydraulic conductance (L), transpiration rate, leaf water status and ABA tissue concentration were measured. KEY RESULTS: None of the treatments (drought, ABA or MTX) changed the leaf water status or tissue ABA concentration. The three treatments caused a decline in the transpiration rate and raised PVPIP2;1 gene expression and PIP1 protein abundance in the leaves. In the roots, only the drought treatment raised the expression of the three PIP genes examined, while at the same time diminishing PIP2 protein abundance and L. On the other hand, ABA raised both root PIP1 protein abundance and L. CONCLUSIONS: The rise of PvPIP2;1 gene expression and PIP1 protein abundance in the leaves of P. vulgaris plants subjected to drought was correlated with a decline in the transpiration rate. At the same time, the increase in the expression of the three PIP genes examined caused by drought and the decline of PIP2 protein abundance in the root tissues were not correlated with any of the parameters measured.     

Changes in the transpiration rate of barley plants infected with powdery mildew in the light and in the dark

Changes in the transpiration rate of intact spring barley plants, cv. “Slovensky dunajsky trh”, were studied separately in the light and in the dark under controlled temperature and illumination, after the infection withErysiphe graminis DC, during an 8 day period of the development of the fungus. In the first stage of pathogenesis, the fungus diminishes water output from the host plants in the light. An opposite phenomenon can be observed in the dark; water output from infected plants in the dark increases sharply mainly in the stage of advanced fructification. Thus, the fungus considerably diminishes the ratio of water output from the host plants in the light to that in the dark.     

Abscisic acid and transpiration rate are involved in the response to boron toxicity in Arabidopsis plants

Boron (B) is an essential microelement for vascular plant development, but its toxicity is a major problem affecting crop yields in arid and semi‐arid areas of the world. In the literature, several genes involved in abscisic acid (ABA) signalling and responses are upregulated in Arabidopsis roots after treatment with excess B. It is known that the AtNCED3 gene, which encodes a crucial enzyme for ABA biosynthesis, plays a key role in the plant response to drought stress. In this study, root AtNCED3 expression and shoot ABA content were rapidly increased in wild‐type plants upon B‐toxicity treatment. The Arabidopsis ABA‐deficient nced3‐2 mutant had higher transpiration rate, stomatal conductance and accumulated more B in their shoots than wild‐type plants, facts that were associated with the lower levels of ABA in this mutant. However, in wild‐type plants, B toxicity caused a significant reduction in stomatal conductance, resulting in a decreased transpiration rate. This response could be a mechanism to limit the transport of excess B from the roots to the leaves under B toxicity. In agreement with the higher transpiration rate of the nced3‐2 mutant, this genotype showed an increased leaf B concentration and damage upon exposure to 5 mM B. Under B toxicity, ABA application decreased B accumulation in wild‐type and nced3‐2 plants. In summary, this work shows that excess B applied to the roots leads to rapid changes in AtNCED3 expression and gas exchange parameters that would contribute to restrain the B entry into the leaves, this effect being mediated by ABA.     

Changes of free and conjugated abscisic acid and phaseic acid in xylem sap of drought-stressed sunflower plants

Abscisic acid (ABA), conjugated abscisic acid, phaseic acid (PA), and conjugated phaseic acid were determined by enzyme-linked immunosorbent assay (ELISA) and gas chromatography (GC) in xylem sap of well-watered and drought-stressed sunflower plants. Conjugated ABA and conjugated PA were determined indirectly after chemical or enzymatic hydrolysis. Conjugated ABA was found to be the predominant ABA metabolite in xylem sap. In xylem sap from well-watered plants at least five, and in sap from drought-stressed plants at least six alkaline hydrolysable ABA conjugates were found. One of them corresponds chromatographically (HPLC) with abscisic acid glucose ester (ABAGE). Under drought conditions the concentrations of ABA, alkaline hydrolysable ABA conjugates, -glucosidase hydrolysable ABA conjugates, PA, and conjugated PA increased. After rewatering the drought-stressed plants, the ABA and the conjugated ABA content decreased. The possible function of the ABA conjugates in the xylem sap as a source of free ABA is discussed.     

Photosynthetic and growth response to fumigation with SO2 at elevated CO2 for C3 and C4 plants

Summary Six early successional plant species with differing photosynthetic pathways (3 C₃ species and 3 C₄ species) were grown at either 300, 600, or 1,200 ppm CO₂ and at either 0.0 or 0.25 ppm SO₂. Total plant growth increased with CO₂ concentration for the C₃ species and varied only slightly with CO₂ for the C₄ species. Fumigation with SO₂ caused reduced growth of the C₃ species at 300 ppm CO₂ but not at the higher concentrations of CO₂. Fumigation with SO₂ reduced growth of the C₄ species at high CO₂ and increased growth at 300 ppm CO₂. Leaf area increased with increasing CO₂ for all plant species. Fumigation with SO₂ reduced leaf area of C₃ plants more at low CO₂ than at high CO₂ while leaf area of C₄ plants was reduced more at high CO₂ than at low CO₂. These results support the notion that C₃ species are more sensitive to SO₂ fumigation than are C₄ species at concentrations of CO₂ equal to that found in normal ambient air. However, the difference in sensitivity to SO₂ between C₃ and C₄ species was found to be reversed at higher concentrations of CO₂. A possible explanation for this reversal based upon differences in stomatal response to elevated CO₂ between C₃ and C₄ species is discussed.     

Changes in the transpiration rate of barley plants cv. “Profesor Schiemann” infected with powdery mildew in light and in darkness

The transpiration rate of intact spring barley plants, cv. Profesor Schiemann resistant to powdery mildew was studied separately in light and in darkness. At the first stage of pathogenesis the fungus temporarily reduced the transpiration rate of the host plants in light (E ₁). The transpiration rate in darkness (E _d) was not influenced in this period and only in the period of formation of necrotic spotsE _d in the host plants was somewhat higher than that in the control plants.E ₁ of the host plants gradually reached the level of the control plants in this period. The fungus considerably reduced the rationE ₁/E _d. The differences in this ratio between infected and control plants increased in the susceptible cultivar but were moderated in the course of necrobiosis.     

Role of superoxide dismutase in defense against SO2 toxicity and an increase in superoxide dismutase activity with SO2 fumigation

The role of superoxide dismutase (SOD) in defense against SO₂toxicity was investigated using leaves of poplar and spinach.Young poplar leaves having five times the SOD of the old leaveswere more resistant to the toxicity of SO₂. Spraying spinachleaves with diethyldithiocarbamate caused a marked loss of SODactivity which resulted in a decrease in their resistance tothe toxic effects of SO₂. The SOD activity in poplar leaveswas increased by fumigation with 0.1 ppm SO₂, and this was moreevident in young leaves than in old ones. The increased SODactivity was inhibited by cyanide. The poplar leaves havinghigh SOD activity induced with SO₂ fumigation were more resistantto 2.0 ppm SO₂ than the control leaves. These findings suggestthat SO₂ toxicity is in part due to the superoxide radical andthat SOD participates in the defense mechanism against SO₂ toxicity. (Received February 12, 1980; )     

Molecular biology and regulation of abscisic acid biosynthesis in plants

The phytohormone abscisic acid (ABA) is involved in seed dormancy and the response to various environmental stresses. Our understanding of the ABA biosynthetic pathway has been increased recently through the use of plant mutants and the cloning of many of the genes encoding for the enzymes involved. C₄₀ Xanthophylls are precursors of ABA and are now known to be derived from isopentenyl phosphate (IPP) synthesized in plastids via a mevalonate-independent pathway. Enzyme reactions downstream of zeaxanthin have recently been reported to be important for the precise regulation of ABA levels. Zeaxanthin epoxidase (ZEP) catalyses the conversion of zeaxanthin to violaxanthin. Changes in ZEP gene expression appear to regulate ABA biosynthesis in seeds and roots, but not in leaves which might be expected considering the important role of epoxy-carotenoids in photosynthesis and photoprotection. The isomerization of the resulting all-trans-violaxanthin to 9-cis-epoxy-carotenoids awaits elucidation. Although 9-cis-epoxy-carotenoid dioxygenase (NCED), which subsequently cleaves the resulting carotenoids could use the 9-cis isomers of both violaxanthin and neoxanthin as substrates in vitro, the in vivo substrates remain to be determined. NCEDs are apparently encoded by multigene families and identification of the various members is required to determine their relative contribution to the regulation of ABA levels. Studies on those already available indicate that their up-regulation upon water stress is compatible with a key role in the modulation of ABA levels. The genes encoding for the enzymes that convert the cleavage product xanthoxin to ABA are not yet known, although recently cloned aldehyde oxidases may act on ABA-aldehyde.     

Altering the sensitivity of strawberry plants to exogenous abscisic acid

Abscisic acid (ABA) causes the closure of stomata. By means of an infrared gas analyzer it was found that changes in the physiological status of Fragaria grandiflora Duch. cv. Red gauntlet, caused by pretreatment with exogenous ABA, decreased the sensitivity to the next dose of the hormone. A similar decrease of sensitivity to ABA may be obtained by shortening the light period. It is proposed that this decrease in sensitivity to ABA may be due to adaptation of guard cells to an excess of ABA.     

Phosphorus concentrations in the leaves of defoliated white clover affect abscisic acid formation and transpiration in drying soil

Increased leaf phosphorus (P) concentration improved the water-use efficiency (WUE) and drought tolerance of regularly defoliated white clover plants by decreasing the rate of daily transpiration per unit leaf area in dry soil. Night transpiration was around 17% of the total daily transpiration. The improved control of transpiration in the high-P plants was associated with an increased individual leaf area and WUE that apparently resulted from net photosynthetic assimilation rate being reduced less than the reductions in the transpiration (27% vs 58%). On the other hand, greater transpiration from low-P plants was associated with poor stomatal control of transpirational loss of water, less ABA in the leaves when exposed to dry soil, and thicker and smaller leaf size compared with high-P leaves. The leaf P concentration was positively related with leaf ABA, and negatively with transpiration rates, under dry conditions ( P < 0.001). However, leaf ABA was not closely related to the transpiration rate, suggesting that leaf P concentration has a greater influence than ABA on the transpiration rates.     

Dark transpiration rate and water deficit as growth limiting factors in alfalfa plants

Growth, dark transpiration rate (DTR) as well as water saturation deficit (WSD) of 30 single plants of 8 alfalfa genotypes growing in experimental field of 50 × 10 cm spacing, in four cuts at early bud stage, were studied. The growth and WSD of genotypes examined were significantly different, the differences in DTR were not significant. The growth of alfalfa plants was in negative correlation with both DTR which reflects non-effective water loss and WSD. Significant negative correlation established between plant growth and its variability shows that fodder productivity in alfalfa genotypes was dependent on growth variability of individual plants. Positive correlations established between WSD or DTR and the growth variability show some of the causes of growth variability.     

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.     

植物脱落酸合成缺陷与反应敏感型突变体

介绍了高等植物体脱落酸生物合成缺陷型突变体,生物合成途径,以及对脱落酸反应超敏感和不敏感的反应型突变体的研究进展。     

Carotenoids and abscisic acid (ABA) biosynthesis in higher plants

Recent research has revealed that abscisic acid (ABA), synthesised in response to water stress, is an apo-carotenoid. Two potential carotenoid precursors, 9'- cis -neoxanthin and 9- cis -violaxanthin, have been identified in light-grown and etiolated leaves, and in roots of a variety of species. Experiments utilizing etiolated Phaseolus vulgaris leaves and deuterium oxide strongly suggest that 9'- cis -neoxanthin, synthesised from all- trans -violaxanthin, is the immediate pre-cleavage precursor of ABA. The cleavage of 9'- cis -neoxanthin, performed by an inducible and specific dioxygenase, is likely to be the rate-limiting step in ABA biosynthesis. Any apocarotenoids formed as by-products of cleavage are probably rapidly degraded by lipoxygenase or related enzymes. After cleavage xanthoxin is converted via ABA-aldehyde to ABA by constitutive enzymes in the cytosol.     

Overproduction of abscisic acid in tomato increases transpiration efficiency and root hydraulic conductivity and influences leaf expansion

Overexpression of genes that respond to drought stress is a seemingly attractive approach for improving drought resistance in crops. However, the consequences for both water-use efficiency and productivity must be considered if agronomic utility is sought. Here, we characterize two tomato (Solanum lycopersicum) lines (sp12 and sp5) that overexpress a gene encoding 9-cis-epoxycarotenoid dioxygenase, the enzyme that catalyzes a key rate-limiting step in abscisic acid (ABA) biosynthesis. Both lines contained more ABA than the wild type, with sp5 accumulating more than sp12. Both had higher transpiration efficiency because of their lower stomatal conductance, as demonstrated by increases in delta(13)C and delta(18)O, and also by gravimetric and gas-exchange methods. They also had greater root hydraulic conductivity. Under well-watered glasshouse conditions, mature sp5 plants were found to have a shoot biomass equal to the wild type despite their lower assimilation rate per unit leaf area. These plants also had longer petioles, larger leaf area, increased specific leaf area, and reduced leaf epinasty. When exposed to root-zone water deficits, line sp12 showed an increase in xylem ABA concentration and a reduction in stomatal conductance to the same final levels as the wild type, but from a different basal level. Indeed, the main difference between the high ABA plants and the wild type was their performance under well-watered conditions: the former conserved soil water by limiting maximum stomatal conductance per unit leaf area, but also, at least in the case of sp5, developed a canopy more suited to light interception, maximizing assimilation per plant, possibly due to improved turgor or suppression of epinasty.     

Inhibition site of the electron transport system in lettuce chloroplasts by fumigation of leaves with SO2

In chloroplasts isolated from SO₂-fumigated leaves at 2.0 ppm,electron flow from water to 2,6-dichloroindophenol (DCIP) wasinhibited, but the electron flow from reduced DCIP to methylviologen was not affected. Neither diphenylcarbazide nor MnCl₂could restore the activity of the DCIP-Hill reaction of SO₂-injuredchloroplasts. Electron flows, from water to ferricyanide orto silicomolybdic acid, were inhibited in a degree similar tothat of the DCIP-Hill reaction. The rate of carotenoid photobleaching in the presence of carbonylcyanide-m-chlorophenylhydrazone was suppressed and paralleledthe inhibition of the DCIP-Hill reaction. In SO₂-injured chloroplasts, the variable part of the fluorescencetransient was diminished, and the fluorescence yield loweredby SO₂ was increased with 3-(3', 4'-dichlorophenyl)-l, l-dimethylurea(DCMU) or more pronouncedly by incubating the sample with sodiumdithionite. However, the yield could not recover to the levelfound in non-fumigated chloroplasts. With SO₂ fumigation, thetime required to reach steady-state level of fluorescence becamelonger in the absence of DCMU, but was not altered in the presenceof DCMU. The pool size of the primary electron acceptors decreasedwith SO₂ fumigation. We concluded that SO₂ inactivated the primaryelectron donor or the reaction center itself. The mode of SO₂action in the electron transport chain is discussed. (Received October 20, 1979; )     

The influence of the transpiration rate on uptake and transport of potassium ions in barley plants

Summary The vegetation points of branches of Caralluma frerei (Asclepiadaceae) were treated with 300, 100 and 30 ppm of crude aflatoxin (36% B₁, 38% G₁, 3% B₂, 2% G₂) and with toxin-free control. Application of 300 and 100 ppm aflatoxin resulted in stop of growth and death of the upper leaves and flower buds. Malformations or wilting was not observed in any case. Branches treated with 30 ppm aflatoxin and with control solution developed normally. It is concluced that under the experimental conditions used aflatoxin has an unspecific toxic effect.     

Cadmium transport by a Cd2+-sensitive and a Cd2+-resistant strain of Bacillus subtilis

Cadmium uptake by a Cd2+-sensitive (1A1) and a Cd2+-resistant mutant (1A1r) strain of Bacillus subtilis was investigated. Uptake of 109Cd2+ was determined for cells of both strains grown in tryptone broth and in broth containing tryptone, yeast extract, and glucose (TYG). The extent of 109Cd2+ uptake by cells of 1A1r was less than by cells of 1A1 under both growth conditions. In both growth media, 109Cd2+ uptake by 1A1 cells demonstrated saturation kinetics and was energy dependent. In both TYG and tryptone broth, 109Cd2+ uptake by 1A1 cells was inhibited by the addition of unlabeled Mn2+. Although lower in magnitude, the kinetics of 109Cd2+ uptake by 1A1r cells were similar to those of 1A1 cells when grown in tryptone broth. However, no obvious saturation kinetics, energy dependence, temperature sensitivity, or inhibition of 109Cd2+ uptake by the addition of unlabeled Mn2+ was observed in 1A1r cells grown in TYG. Differential Mn2+ accumulation by 1A1r cells in TYG and tryptone broth correlated with differential 109Cd2+ uptake by 1A1r cells in these media.