Stomatal sensitivity to abscisic acid following water deficit stress

Short-and medium-term stresses (1 and 24 h, respectively) wereapplied to detached leaves of Commelina communis L., resultingin both cases in a final leaf cell water potential (     

A simplified purification method for the analysis of abscisic acid

A fast and convenient method is given for the purification of abscisic acid (ABA) in plant extracts. Using a dual pH elution system on Sep-Pak C18 reverse phase prepacked cartridges, abscisic acid appears in the third eluent (32% methanol pH 8.0). The cartridges can be regenerated for multiple reuse.     

Transport of abscisic acid

The transport of abscisic acid in cotyledonary petiole sectionsof cotton seedlings was investigated using (2-¹⁴C)-ABA in adonor-receiver system. Using the intercept technique, a transportvelocity of 22.4 mm/hr was determined. This velocity was foundto be independent of the length of the petiole or of the concentrationof ABA applied. In a 24-hr period 70% of the applied ABA movedthrough the tissue sections. No polarity was found in any ofthe studies. The capacity of the transport system decreased with increasingage of the tissue and with increasing time of pre-exposure toABA. When metabolism of the tissue was restricted by cold temperature,low-O₂ tension or DNP treatment, transport was inhibited 80to 98%.The distribution of ABA in the petiole during transportwas also investigated. The results are discussed in relationto the well-studied transport of IAA and in relation to thephysiologicalrole of ABA. ¹ Present address: Stanford Research Institute-Irvine, 19722Jamboree Blvd., Irvine, Calif. 92664, U.S.A. (Received November 30, 1970; )     

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.     

Effects of patchy stomatal closure on gas exchange measurements following abscisic acid treatment

Gas exchange data and images of leaf fluorescence were collected concurrently as stomata responded to abscisic acid (ABA) application. When 10^?5kmolm^?3 ABA was applied to the transpiration stream in a short pulse, stomatal conductance (g_s), photosynthesis (A) and intercellular CO2 concentration (C_i) decreased rapidly after a short lag period and became approximately constant after 2h. There was an apparent reduction in the A versus c1 relationship as stomata closed, but the data returned to the A versus C1 curve while stomatal conductance was constant or slowly rising during the second hour after ABA treatment. Larger amounts of ABA administered during the pulse caused larger deviations from the A versus c1 relationship. When 10^?7kmolm^?3 ABA was applied continuously through the transpiration stream, gs, A and Cⁱ decreased, but there was no substantial deviation from the A versus c{ curve. Fluorescence images were patchy as stomata closed for all experiments, but became slowly more uniform during the time that gas exchange was returning to the A versus Cj curve. The distribution of con-ductance among patches was not bimodal, and larger devi-ations from the A versus ct curve had greater ranges of pixel values and more pixel values representing low values of Cj during stomatal closure than did experiments show-ing small or no deviation. Estimates of A and gs from fluo-rescence images compared favourably with measured val-ues in most cases, suggesting that the patchy distributions of fluorescence were caused by patchy distributions of stomatal conductance and that apparent reductions in the A versus ct relationship were the result of these patchy stomatai distributions and not direct effects of ABA on mesophyll functioning. The data show that stomatal patches can be temporary and that patchiness may not be reflected in gas exchange data if the range of stomatal con-ductances is not large. These observations may explain some of the discrepancies among previous studies concerning the effect of ABA on the A versus Cⁱ relationship.     

Transpiration-inhibiting abscisic acid analogs

Synthetic analogs of abscisic acid (ABA) and their inhibiting effect on transpiration rates of detached barley leaves are presented. By systematically varying the carbon skeleton of ABA, the influence of structural changes on biological activity was investigated. The results show that a properly substituted cyclohexane unit and a six-carbon side chain seem to be indispensable for high ABA-like activity, whereas the oxidation state of the terminal carbon atom in the side chain appears to be less essential. Thus, synthetic compounds have been created that exhibit biological activities comparable both in type and strength with ABA itself. On the basis of molecular models, a hypothesis of the geometric arrangement of essential substructural units is proposed.     

Transpiration-inhibiting abscisic acid analogs

Synthetic analogs of abscisic acid (ABA) and their inhibiting effect on transpiration rates of detached barley leaves are presented. By systematically varying the carbon skeleton of ABA, the influence of structural changes on biological activity was investigated. The results show that a properly substituted cyclohexane unit and a six-carbon side chain seem to be indispensable for high ABA-like activity, whereas the oxidation state of the terminal carbon atom in the side chain appears to be less essential. Thus, synthetic compounds have been created that exhibit biological activities comparable both in type and strength with ABA itself. On the basis of molecular models, a hypothesis of the geometric arrangement of essential substructural units is proposed.     

Chemical biology of abscisic acid

Chemical biology is a discipline that utilizes chemicals to elucidate biological mechanisms and physiological functions. Various abscisic acid (ABA) derivatives have revealed the structural requirement for the perception by ABA receptors while biotin or caged derivatives of ABA have disclosed the localization of several ABA-binding proteins. Recently, selective ABA agonist has been used to identify ABA receptors. Furthermore, ABA biosynthesis and catabolic inhibitors have contributed to the identification of new ABA functions in plant growth and development. The physiological function of ABA in non-plant organisms has gradually been revealed. In this review, we discuss the development of small bioactive chemicals and their significance in ABA research.     

Cuticular penetration of abscisic acid

Summary Penetration of 2-¹⁴C abscisic acid (ABA) through enzymatically isolated cuticles from tomato fruit and from the upper epidermis of apricot, pear and orange leaves was assessed. Penetration was linear with time, greater as the undissociated than the dissociated ion, and greater through dewaxed than non-dewaxed cuticles. Significantly less (3–6 times) (2-¹⁴C)ABA penetrated the tomato fruit cuticle than NAA or 2,4-D. The leaf cuticles were less permeable than the tomato fruit cuticle. There was no evidence that the ABA was altered during transfer across the cuticle.     

Control of abscisic acid synthesis

The abscisic acid (ABA) biosynthetic pathway involves the formation of a 9-cis-epoxycarotenoid precursor. Oxidative cleavage then results in the formation of xanthoxin, which is subsequently converted to ABA. A number of steps in the pathway may control ABA synthesis, but particular attention has been given to the enzyme involved in the oxidative cleavage reaction, i.e. 9-cis-epoxycarotenoid dioxygenase (NCED). Cloning of a gene encoding this enzyme in maize was first reported in 1997. Mapping and DNA sequencing studies indicated that a wilty tomato mutant was due to a deletion in the gene encoding an enzyme with a very similar amino acid sequence to this maize NCED. The potential use of this gene in altering ABA content will be discussed together with other genes encoding ABA biosynthetic enzymes.     

反相高效液相色谱法测定平邑

用高效液相色谱法（ＨＰＬＣ）分析平邑甜茶内源ＡＢＡ、ＩＡＡ,选用Ｓｐｈｅｒｉｓｏｒｂ Ｃ１８反相柱,用ＵＶ检测器进行检测,使ＡＢＡ、ＩＡＡ得到很好的分离,测定方法迅速简便,流动相为甲醇、乙酸和水,系统研究了ｐＨ值、纯化方法对测定结果的影响,确定适合平邑甜茶ＡＢＡ、ＩＡＡ提取的分离条件。方法的精密度为：变异系数分别为２．８３和２．７９,最低检出限（信噪比＝２）分别为３．２７和３．２６ｎｍｏｌ／Ｌ,线     

An improved method for the direct chromatographic resolution of abscisic acid enantiomers

An improved method for the direct chromatographic resolution of abscisic acid enantiomers using the commercially available Whelk-O 1 chiral stationary phase is presented. Previously reported strategies utilized in the chromatographic separation of abscisic acid enantiomers are summarized, and conditions for analytical and semipreparative separations using the Whelk-O 1 chiral stationary phase are described. This method offers the advantages of rapid analysis time and greatly increased capacity, allowing the resolution of more than 6 mg in 25 min using an analytical column (4.6 mm i.d. × 25 cm length). © 1993 Wiley-Liss, Inc.     

Abscission-promoting activities of abscisic acid and five abscisic acid analogs in cotton seedlings and explants

The abscission-promoting activities of abscisic acid (ABA) and 5 ABA analogs were examined in cotton (Gossypium hirsutum L. cv LG102) seedlings and cotyledonary node explants. The analogs tested included a series of acetylenic derivatives that differ in the oxidation state of the C-1 atom, a 2,3 dihydro-derivative of ABA and a 2,3 dihydro-derivative of an acetylenic analog with a C-1 carboxyl moiety. ABA and all five analogs were active in stimulating petiole abscission in explants. Following treatment with 100,µM ABA or analog, 50% abscission of explants was observed after 29 h and complete abscission occurred within 40 h. With one exception, none of the treatments resulted in an increase in explant ethylene production. Pretreatment of the explants with the ethylene antagonist silver thiosulfate completely abolished the abscission-promoting activities of ABA and all of the analogs. Daily application of ABA or any of the analogs had no effect on cotyledon abscission in intact seedlings. The implications of the results with respect to the development of a commercial ABA-like regulator as well as to ABA structure-activity studies are discussed.Mention of trademark or proprietary product does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products that may also be suitable.     

Determination of endogenous abscisic acid levels in immature cereal embryos during in vitro culture

Levels of endogenous abscisic acid (ABA) in immature wheat (Triticum aestivum cv. Timmo) and barley (Hordeum vulgare cv. Golden Promise) embryos have been determined by enzyme-linked immunosorbent assay. Embryos of both cereal species showed an increase in ABA content during development on the parent plant. Immature embryos were excised and cultured in vitro on nutrient media that led to precocious germination or on media containing 9% (w/v) mannitol that maintained their developmental arrest. Barley and wheat embryos responded to these culture conditions in an identical manner with respect to changes in morphology, fresh weight, protein and lectin content. However, in complete contrast, the ABA content of barley embryos increased by an order of magnitude during culture on mannitol, whereas that of wheat embryos showed no significant change. The results are discussed within the context of the role of ABA in the regulation of embryo development.Abbreviations ABA abscisic acid - BGA barley-germ agglutinin - dpa days post anthesis - ELISA enzyme-linked immunosorbent assay - GC-MS gas chromatography-mass spectrometry - WGA wheat-germ agglutinin     

Ultrastructural and cytochemical aspects of induced apogamy following abscisic acid pre-treatment of secondary moss protonema

Abscisic acid (ABA) treatment of secondary protonema of Physcomitrium pyriforme Brid in the presence of sucrose does not prevent cell division but results in shorter cells with vesicular cytoplasm and an accumulation of lipid. When transferred to sucrose medium without ABA and with low irradiance isodiametric intercalary cells are cut off which give rise to apogamous sporophytes either directly or after the formation of a small amount of callus. The organization of the cells leading up to the apogamous sporophyte is described. The cells initiating the sporophyte develop dense cytoplasm and the walls become labyrinthine and callosed, but they do not form any recognizable placenta. It is proposed that labyrinthine walls are a consequence of a perturbation of cell wall metabolism as growth changes from gametophytic to sporophytic. The use of the term transfer cell for this kind of cell is questioned and the need for a causal approach to the investigation of labyrinthine walls is stressed.     

Increased abscisic acid sensitivity and drought tolerance of Arabidopsis by overexpression of poplar abscisic acid receptors

Plant Cell, Tissue and Organ Culture (PCTOC) - Abscisic acid (ABA), a key plant hormone that regulates plant growth development and stress response, is recognized and bound by ABA Receptor...     

Inhibitors of abscisic acid 8'-hydroxylase

Structural analogues of the phytohormone (+)-abscisic acid (ABA) have been synthesized and tested as inhibitors of the catabolic enzyme (+)-ABA 8'-hydroxylase. Assays employed microsomes from suspension-cultured corn cells. Four of the analogues [(+)-8'-acetylene-ABA, (+)-9'-propargyl-ABA, (-)-9'-propargyl-ABA, and (+)-9'-allyl-ABA] proved to be suicide substrates of ABA 8'-hydroxylase. For each suicide substrate, inactivation required NADPH, increased with time, and was blocked by addition of the natural substrate, (+)-ABA. The most effective suicide substrate was (+)-9'-propargyl-ABA (K(I) = 0.27 microM). Several analogues were competitive inhibitors of ABA 8'-hydroxylase, of which the most effective was (+)-8'-propargyl-ABA (K(i) = 1.1 microM). Enzymes in the microsomal extracts also hydroxylated (-)-ABA at the 7'-position at a low rate. This activity was not inhibited by the suicide substrates, showing that the 7'-hydroxylation of (-)-ABA was catalyzed by a different enzyme from that which catalyzed 8'-hydroxylation of (+)-ABA. Based on the results described, a simple model for the positioning of substrates in the active site of ABA 8'-hydroxylase is proposed. In a representative physiological assay, inhibition of Arabidopsis thaliana seed germination, (+)-9'-propargyl-ABA and (+)-8'-acetylene-ABA exhibited substantially stronger hormonal activity than (+)-ABA itself.     

A radioimmunoassay for abscisic acid

We have developed a radioimmunoassay (RIA) for abscisic acid (ABA) in the 0.1 ng to 2.5 ng range. Antibodies were obtained from rabbits immunized with ABA bound via its carboxyl group to bovine serum albumin. Cross-reactivity studies indicate that ABA esters are completely cross-reactive with ABA, while trans, trans abscisic acid (t-ABA) phaseic acid (PA) and dihydrophaseic acid (DPA) have much lower but significant cross-reactivities. Purification methods which reduce the levels of cross-reacting substances are described.Abbreviations RIA radioimmunoassay - DPA 4-dihydrophaseic acid - PA phaseic acid - GC gas chromatography - HPLC high performance liquid chromatography - TLC thin-layer chromatography - BSA bovine serum albumin - ABA abscisic acid - t-ABA trans, trans abscisic acid - IAA indoleacetic acid