Molecular requirements for abscisic acid activity in two bioassay systems

Summary Twenty-two analogues of ABA have been tested in the Avena coleoptile and lettuce germination bioassays. Ten of these analogues were considerably more active than ABA itself as inhibitors of lettuce germination, but in the Avena coleoptile assay their activity never significantly exceeded that of ABA. The molecular requirements for activity also differ in the two assay systems, although the presence of the ring double bond is a requirement of both.     

Does Xylem Sap ABA Control the Stomatal Behaviour of Water-Stressed Sycamore (Acer pseudoplatanus L.) Seedlings?

Sycamore seedlings were grown with their root systems dividedequally between two containers. Water was withheld from onecontainer while the other container was kept well-watered. Effectsof soil drying on stomatal behaviour, shoot water status, andabscisic acid (ABA) concentration in roots, xylem sap and leaveswere evaluated. At 3 d, root ABA in the drying container increased significantly,while the root ABA in the unstressed container of the same plantsdid not differ from that of the control. The increase in rootABA was associated with the increase in xylem sap ABA and withthe decrease in stomatal conductance without any significantperturbation in shoot water status. At 7 d, despite the continuous increase in root ABA concentration,xylem sap ABA showed a marked decline when soil water contentwas depleted below 013 g g^–1. This reduction in xylemsap ABA coincided with a partial recovery of stomatal conductance.The results indicate that xylem sap ABA is a function of rootABA as well as the flow rate of water from roots to shoots,and that this ABA can be a sensitive indicator to the shootof the effect of soil drying. Key words: Acer pseudoplatanus L., soil drying, stomatal behaviour, xylem sap ABA     

Amelioration of chilling-induced water stress by abscisic Acid-induced changes in root hydraulic conductance

Pretreatment of soybean (Glycine max L. var Ransom) root systems with abscisic acid (ABA) ameliorates the deleterious effect of low temperatures on root hydraulic conductance. ABA treatment of root systems subsequently chilled to 10°C with shoots at 25°C resulted in higher leaf water potentials and lower stomatal resistances. If the root systems are left at 25°C, ABA causes stomatal closure. Membrane alterations are suggested as a mechanism for the ABA action in plant response to chilling stress.     

Root water flow and leaf stomatal conductance in aspen (Populus tremuloides) seedlings treated with abscisic acid

Exogenous abscisic acid (ABA) applied to the roots and excised shoots of aspen (Populus tremuloides Michx.) inhibited stomatal conductance. However, the effect of ABA on stomatal conductance was more pronounced in the excised shoots compared with the intact seedlings. Approximately 10% of the ABA concentration applied to the roots was found in the xylem exudates of root systems exposed to a hydrostatic pressure of 0.3 MPa. A similar concentration of ABA applied to the excised shoots produced a faster and greater reduction of stomatal conductance. ABA applied to the roots had no effect on root steady-state flow rate over the 5-h experimental period. Moreover, pre-incubating root systems of intact seedlings for 12 h with 5 x 10(-5) M ABA did not significantly reduce volume flow density. Similarly, ABA had no effect on root hydraulic conductivity and the activation energy of root water flow rates.     

Biological and chemical properties of the epidioxide isomer of abscisic Acid and its rearrangement products

The growth inhibitory activity of the epidioxide (II), a precursor in the synthesis of abscisic acid (ABA), has been confirmed with additional assay systems. Under physiological conditions the epidioxide is rearranged to give ABA and an isomer of ABA which has probably the structure V. This major product has very low, if any, biological activity. The biological activity of the epidioxide is explained by its partial conversion (about 20%) to ABA. The reaction rate was enhanced by heavy metal ions and decreased by EDTA. At pH 12.5, the decomposition of the epidioxide is slower than it is near neutrality and ABA is the predominant product. In the biological systems studied the activity of the epidioxide can be accounted for by nonenzymatic conversion to ABA.     

Sensitivity of Stomata to Abscisic Acid (An Effect of the Mesophyll)

The effects of added abscisic acid (ABA) on the stomatal behavior of Commelina communis L. were tested using three different systems. ABA was applied to isolated epidermis or to leaf pieces incubated in the light in bathing solutions perfused with CO2-free air. ABA was also fed to detached leaves in a transpiration bioassay. The apparent sensitivity of stomata to ABA was highly dependent on the method used to feed ABA. Stomata of isolated epidermis were apparently most sensitive to ABA, such that a concentration of 1 [mu]M caused almost complete stomatal closure. When pieces of whole leaves were floated on solutions of ABA of the same concentration, the stomata were almost completely open. The same concentration of ABA fed through the midrib of transpiring detached leaves caused an intermediate response. These differences in stomatal sensitivity to added ABA were found to be a function of differences in the ABA concentration in the epidermes. Comparison of the three application systems suggested that, when leaf pieces were incubated in ABA or fed with ABA through the midrib, accumulation of ABA in the epidermes was limited by the presence of the mesophyll. Even bare mesophyll incubated in ABA solution did not accumulate ABA. Accumulation of radioactivity by leaf pieces floated on [3H]ABA confirmed ABA uptake in this system. Experiments with tetcyclacis, an inhibitor of phaseic acid formation, suggested that rapid metabolism of ABA in mesophyll can have a controlling influence on ABA concentration in both the mesophyll and the epidermis. Inhibition of ABA catabolism with tetcyclacis allows ABA accumulation and increases the apparent sensitivity of stomata to applied ABA. The results are discussed in the context of an important role for ABA metabolism in the regulation of stomatal behavior.     

The role of abscisic acid in plant tissue culture: a review of recent progress

Abscisic acid (ABA) plays a significant role in the regulation of many physiological processes of plants. It is often used in tissue culture systems to promote somatic embryogenesis and enhance somatic embryo quality by increasing desiccation tolerance and preventing precocious germination. ABA is also employed to induce somatic embryos to enter a quiescent state in plant tissue culture systems and during synthetic seed research. Application of exogenous ABA improves in vitro conservation and the adaptive response of plant cell and tissues to various environmental stresses. ABA can act as anti-transpirant during the acclimatization of tissue culture-raised plantlets and reduces relative water loss of leaves during the ex vitro transfer of plantlets even when non-functional stomata are present. This review focuses on the possible roles of ABA in plant tissue culture and recent developments in this area.     

水分胁迫积累的ABA诱导抗氧化防护系统的信号级联

水分胁迫是限制植物生长发育的主要胁迫因子之一。植物通过感受刺激,产生和传递信号、启动多种防御机制对水分胁迫做出响应和适应。脱落酸(ABA)作为一种重要的植物体内胁迫激素,参与了许多这样的反应。研究表明,ABA增强植物水分胁迫的忍耐力与ABA诱导的抗氧化剂防护系统有关;且细胞溶质Ca2 ([Ca2 ]i)、活性氧(ROS)等许多第二信使参与了ABA诱导的信号转导过程。本文就这些信号分子在水分胁迫积累的内源ABA诱导的抗氧化剂防护系统中的作用作一综述。     

Abscisic acid in lichens: variation, water relations and metabolism

For the first time the well known drought stress hormone abscisic acid, which is involved in regulating processes increasing desiccation tolerance in many plant systems, was analysed in lichens. ABA was detected in all 26 species investigated. In contrast to higher plants and liverworts, the ABA content increased after hydration of air dry lichen thalli and decreased in desiccating lichen material. Experiments with Baeomyces rufus (Huds.) Rebent indicated that the mycobiont might be the major site of ABA biosynthesis. After incubation of hydrated lichen thalli with radioactive ABA for up to 72 h no metabolism to phaseic acid and dihydrophaseic acid could be detected. Fluctuations of internal ABA might be a result of ABA release to the external medium.     

Involvement of plasma-membrane NADPH oxidase in abscisic acid- and water stress-induced antioxidant defense in leaves of maize seedlings

The roles of the plasma-membrane (PM) NADPH oxidase in abscisic acid (ABA)- and water stress-induced antioxidant defense were investigated in leaves of maize ( Zea mays L.) seedlings. Treatment by exogenous ABA (100 micro M ABA) or osmotic stress (-0.7 MPa induced by polyethylene glycol) significantly increased the activity of the PM NADPH oxidase, the production of leaf O(2)(-), the activities of several antioxidant enzymes (superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase), and the contents of antioxidant metabolites (ascorbate and reduced glutathione). Pretreatment with three different inhibitors of NADPH oxidase (diphenylene iodonium, imidazole and pyridine) or an inhibitor of ABA biosynthesis (tungstate) reduced the increase in the activity of the PM NADPH oxidase and the production of leaf O(2)(-), and the capacity of antioxidant defense systems mediated by ABA. The inhibitory effects above caused by tungstate were reversed by exogenous ABA. These data indicate that NADPH oxidase is involved in the ABA-induced production of active oxygen species (AOS), and our results depict a minimal chain of events initiated by water stress-induced ABA accumulation, which then triggers the production of AOS by membrane-bound NADPH oxidase, resulting in the induction of antioxidant defense systems against oxidative damage in plants.     

Dynamic analysis of ABA accumulation in relation to the rate of ABA catabolism in maize tissues under water deficit

The plant hormone abscisic acid (ABA) accumulates in plant tissues which experience water deficit (stress ABA). This study analysed its accumulation as a function of both synthesis and catabolism in maize tissues. By following the disappearance of the stress ABA when ABA synthesis was blocked by nordihydroguaiaretic acid (NDGA), the rate of the catabolism of stress ABA was determined. When compared with the catabolic rate of baseline (non-stress) ABA, stress ABA showed a catabolic rate >11 times higher. With such an elevated catabolic rate, it is proposed that the xanthophyll precursor pool may not be able to sustain the ABA accumulation, and such a proposition has been substantiated by further experiments where fluridone is used to limit the availability of upstream ABA precursors. When fluridone was used, stress ABA accumulation could only be sustained for a few hours, i.e. approximately 5 h for leaf and 1 h for root tissues. In detached roots, stress ABA accumulation could not be sustained even if fluridone was not used, suggesting that stress ABA accumulation in root systems requires the continuous import of ABA precursors from the shoots. Such an assumption was substantiated by the observation that defoliation or shading significantly reduced ABA accumulation in intact roots. The present study suggests that ABA catabolism is rapid enough to play an important role in the regulation of ABA accumulation.     

Movement of Abscisic Acid into the Apoplast in Response to Water Stress in Xanthium strumarium L

The effect of water stress on the redistribution of abcisic acid (ABA) in mature leaves of Xanthium strumarium L. was investigated using a pressure dehydration technique. In both turgid and stressed leaves, the ABA in the xylem exudate, the `apoplastic' ABA, increased before `bulk leaf' stress-induced ABA accumulation began. In the initially turgid leaves, the ABA level remained constant in both the apoplast and the leaf as a whole until wilting symptoms appeared. Following turgor loss, sufficient quantities of ABA moved into the apoplast to stimulate stomatal closure. Thus, the initial increase of apoplastic ABA may be relevant to the rapid stomatal closure seen in stressed leaves before their bulk leaf ABA levels rise.

Following recovery from water stress, elevated levels of ABA remained in the apoplast after the bulk leaf contents had returned to their prestress values. This apoplastic ABA may retard stomatal reopening during the initial recovery period.

     

Abscisic acid in the plants-pathogen interaction

Information available concerning the role of ABA in the interaction between plants and pathogenic microorganisms allows a conclusion that this phytohormone is required for plant defense. For the development of plant resistance, short-term increases in the ABA level are of importance during the early stages of plant interaction with pathogens, which trigger anti-stress programs in plants, primarily related to the synthesis of callose. At the same time, high ABA concentrations maintained for a long time reduce efficiency of defense systems controlled by salicylic and jasmonic acids and ethylene. ABA was shown to suppress expression of some genes of defense proteins, including those involved in the synthesis and metabolism of phenolic compounds and lignin. ABA is evidently involved in plant defense mechanisms against pathogens as a regulatory element.     

Action of natural abscisic acid precursors and catabolites on abscisic acid receptor complexes

The phytohormone abscisic acid (ABA) regulates stress responses and controls numerous aspects of plant growth and development. Biosynthetic precursors and catabolites of ABA have been shown to trigger ABA responses in physiological assays, but it is not clear whether these are intrinsically active or whether they are converted into ABA in planta. In this study, we analyzed the effect of ABA precursors, conjugates, and catabolites on hormone signaling in Arabidopsis (Arabidopsis thaliana). The compounds were also tested in vitro for their ability to regulate the phosphatase moiety of ABA receptor complexes consisting of the protein phosphatase 2C ABI2 and the coreceptors RCAR1/PYL9, RCAR3/PYL8, and RCAR11/PYR1. Using mutants defective in ABA biosynthesis, we show that the physiological activity associated with ABA precursors derives predominantly from their bioconversion to ABA. The ABA glucose ester conjugate, which is the most widespread storage form of ABA, showed weak ABA-like activity in germination assays and in triggering ABA signaling in protoplasts. The ABA conjugate and precursors showed negligible activity as a regulatory ligand of the ABI2/RCAR receptor complexes. The majority of ABA catabolites were inactive in our assays. To analyze the chemically unstable 8'- and 9'-hydroxylated ABA catabolites, we used stable tetralone derivatives of these compounds, which did trigger selective ABA responses. ABA synthetic analogs exhibited differential activity as regulatory ligands of different ABA receptor complexes in vitro. The data show that ABA precursors, catabolites, and conjugates have limited intrinsic bioactivity and that both natural and synthetic ABA-related compounds can be used to probe the structural requirements of ABA ligand-receptor interactions.     

Structural Requirements of Abscisic Acid (ABA) and its Impact on Water Flow During Radial Transport of ABA Analogues Through Maize Roots

The effect of (+) (ABA) and (?)-abscisic acid and nine ABA metabolites, precursors or derivatives on radial water movement through maize roots, was investigated using a suction technique (Freundl and others 1998). (+)-ABA, (+)- and (?)-abscisyl aldehyde, (+)-8?-hydroxymethyl ABA, (+)-8?-methylene, and (+)-8?-acetylene ABA stimulated radial water transport. (?)-ABA, phaseic acid, and (+)-8?-acetylene methyl ABA were ineffective. ELISA analysis for ABA detected and apparent increase of free ABAxyl in xylem sap of excised root systems that were perfused with either (+)-abscisyl aldehyde, (+)-8?-methylene, (+)8?-acetylene-ABA, or ABA-glucose ester. The analogues (+)-8?-hydroxymethyl ABA and (?)-abscisyl aldehyde passed the cortex of maize roots without changing the ABAxyl. The data from this study permit conclusions about the structural requirements for hormonal regulation of hydraulic conductivity.