Abscisic acid biosynthesis in roots

The pathway of water-stress-induced abscisic acid (ABA) biosynthesis in etiolated and light-grown leaves has been elucidated (see A.D. Parry and R. Horgan, 1991, Physiol. Plant. 82, 320–326). Roots also have the ability to synthesise ABA in response to stress and it was therefore of interest to examine root extracts for the presence of carotenoids, including those known to be ABA precursors in leaves. All-trans- and 9-cis-neoxanthin, all-trans- and 9-cis-violaxanthin, antheraxanthin (all potential ABA precursors), lutein and -carotene were identified on the basis of absorbance spectra, reactions with dilute acid, retention times upon high-performance liquid chromatography and by comparison with leaf carotenoids that had been analysed by mass spectrometry. The source of the extracted carotenoids was proved to be root tissue, and not contaminating compost or leaf material. The levels of total carotenoids in roots varied between 0.03–0.07% of the levels in light-grown leaves (Arabidopsis thaliana (L.) Heynh, Nicotiana plumbaginifolia Viv., Phaseolus vulgaris L. and Pisum sativum L.) up to 0.27% (Lycopersicon esculentum Mill.). The relative carotenoid composition was very different from that found in leaves, and varied much more between species. All-trans-neoxanthin and violaxanthin were the major carotenoids present (64–91 % of the total), but while Lycopersicon contained 67–80% all trans-neoxanthin, Phaseolus, Pisum and Zea mays L. contained 61–79% all-trans-violaxanthin. Carotenoid metabolism also varied between species, with most of the carotenoids in older roots of Phaseolus being esterified. Roots and leaves of the ABA-deficient aba mutant of Arabidopsis had reduced epoxy-xanthophyll levels compared to the wild-type.Abbreviations ABA abscisic acid - r.p.HPLC reversed-phase high performance liquid chromatography The authors would like to thank Dr. B.H. Davies for helpful discussions and Mrs. A.F. Rees for her excellent technical assistance. A.D.P. was supported by a grant from the Agricultural and Food Research Council, from whom funds were also obtained to purchase the HPLC-photodiode-array detector.     

Abscisic acid perception and signaling transduction in strawberry: A model for non-climacteric fruit ripening

On basis of fruit differential respiration and ethylene effects, climacteric and non-climacteric fruits have been classically defined. Over the past decades, the molecular mechanisms of climacteric fruit ripening were abundantly described and found to focus on ethylene perception and signaling transduction. In contrast, until our most recent breakthroughs, much progress has been made toward understanding the signaling perception and transduction mechanisms for abscisic acid (ABA) in strawberry, a model for non-climacteric fruit ripening. Our reports not only have provided several lines of strong evidences for ABA-regulated ripening of strawberry fruit, but also have demonstrated that homology proteins of Arabidopsis ABA receptors, including PYR/PYL/RCAR and ABAR/CHLH, act as positive regulators of ripening in response to ABA. These receptors also trigger a set of ABA downstream signaling components, and determine significant changes in the expression levels of both sugar and pigment metabolism-related genes that are closely associated with ripening. Soluble sugars, especially sucrose, may act as a signal molecular to trigger ABA accumulation through an enzymatic action of 9-cis-epoxycarotenoid dioxygenase 1 (FaNCED1). This mini-review offers an overview of these processes and also outlines the possible, molecular mechanisms for ABA in the regulation of strawberry fruit ripening through the ABA receptors.     

高等植物脱落酸生物合成的酶调控

高等植物ABA的生物合成开始于细胞质内的甲瓦龙酸 (MVA)或位于叶绿体内的丙酮酸_硫胺素焦磷酸 (TPP) ,经一系列反应最后在质体或胞质中形成的。除胁迫或植物发育中生理变化引起的诱导外 ,ABA的合成还受到一系列酶的调控 ,其中 ,玉米黄质环氧化酶 (ZE) ,9_顺环氧类胡萝卜素双加氧酶(NCED)和醛氧化酶 (AO)可能起到重要的调节作用。本文介绍近年来ABA生物合成酶调控的研究进展。     

植物激素ABA调控植物根系生长的研究进展

脱落酸（abscisic acid,ABA）是一种重要的植物激素,在调控种子发育、种子休眠与萌发、抑制生长、促进落花落果、参与植物应对外界环境胁迫等过程中发挥着重要的生理功能。ABA还能与其他植物激素（如生长素、乙烯等）互作进而精细调控植物根系的生长。本文以模式植物拟南芥（Arabidopsis thaliana（L.）Heynh）为主要对象,对近年来国内外在ABA调控植物根系生长方面的研究成果、ABA与其他植物激素（如GA等）互作调控根系生长及调控非生物逆境下根系发育的机理等进行综述,并对其未来的研究方向进行了展望。     

Abscisic acid physiology and biosynthesis in higher plants

Abscisic acid (ABA) has been postulated to modulate several aspects of plant growth and development. While it is tempting to attribute changes in growth and development to a specific hormone such as ABA, the reality is that these processes are complex and poorly understood. Since there is so little known about basic biochemical events that occur during growth and development, it is difficult ot unambiguously assign a role for ABA in any process. Becuse of this, many of the cited effects of ABA on growth and development have not been conclusively demonstrated. Howver, it is clear that ABA has a function in ameliorating water-stress and preventing vivipary. The roles of ABA in bud dormancy and growth still remain unclear. With the use of biosynthesis inhibitors and mutants which block ABA accumulation, it has been shown that ABA does not play a role in gravitropism.
Knowledge of how the levels of any particular growth regulator are modulated is essential for the understanding of its physiology. The use of mutants, inhibitors and heavy isotopes suggests that ABA may be derived from a carotenoid rather than directly from farnesyl pyrophosphate (FPP), and that the cleavage of a carotenoid is the rate limiting step. However, the relative contribution of each pathway (and the role of xanthoxin) in ABA biosynthesis remain unknown.     

Abscisic acid is a negative regulator of root gravitropism in Arabidopsis thaliana

The plant hormone abscisic acid (ABA) plays a role in root gravitropism and has led to an intense debate over whether ABA acts similar to auxin by translating the gravitational signal into directional root growth. While tremendous advances have been made in the past two decades in establishing the role of auxin in root gravitropism, little progress has been made in characterizing the role of ABA in this response. In fact, roots of plants that have undetectable levels of ABA and that display a normal gravitropic response have raised some serious doubts about whether ABA plays any role in root gravitropism. Here, we show strong evidence that ABA plays a role opposite to that of auxin and that it is a negative regulator of the gravitropic response of Arabidopsis roots.     

Abscisic acid receptors: multiple signal-perception sites

BACKGROUND AND AIMS: The phytohormone abscisic acid (ABA) plays a vital role in various aspects of plant growth and development and in adaptation of plants to various environmental stresses. Cell response to ABA is initiated by ABA perception with a cell receptor. Recently, three distinct ABA receptors have been identified, opening a door to uncover the initial events of ABA signal transduction. The aim of this Botanical Briefing is to present a perspective of the ABA receptors identified. SCOPE: This Briefing offers an introduction to the three ABA receptors identified and an analysis of the complexity and multiplicity of ABA receptors, and provides some viewpoints on future research.     

In vivo and in vitro swelling of cell walls during fruit ripening

Swelling properties of the cell walls of nine temperate fruit species, selected for their different ripening and textural characteristics, were studied during ripening. Cell wall swelling was examined in intact fruit using microscopy techniques and in vitro, using cell wall material isolated from fruit tissue. In fruit which ripened to a soft melting texture (persimmon, avocado, blackberry, strawberry, plum), wall swelling was pronounced, particularly in vitro. In-vivo swelling was marked only in avocado and blackberry. Fruit which ripened to a crisp, fracturable texture [apple (two cultivars), nashi pear, watermelon] did not show either in-vivo or in-vitro swelling of the cell wall. There was a correlation between swelling and the degree of pectin solubilisation, suggesting that wall swelling occurred as a result of changes to the viscoelastic properties of the cell wall during pectin solubilisation. Chemical and enzymatic removal of pectin from kiwifruit cell wall material supported the idea that swelling is associated with movement of water into voids left in the cellulose-hemicellulose network by the solubilised pectin. However, the results also suggested that swelling in vivo was more complex than this, and that the physicochemical changes which led to swelling included other elements of cell wall modification involving the site and mechanism of pectin solubilisation and-or the cellulose-xyloglucan complex. Received: 28 January 1997 / accepted: 11 March 1997     

真菌中脱落酸的代谢及定量分析技术

综述了各种脱落酸产生真菌的生物学特征及其不同的生物合成代谢途径,并对脱落酸的定量分析技术作了简要介绍。     

Abscisic acid biosynthesis during corn embryo development

In this study we examined the biosynthesis of abscisic acid (ABA) by developing corn (Zea mays L.) embryos. Three comparisons were made: ABA biosynthesis in embryos isolated from kernels grown in vitro with those grown in the field; the developmental profile of ABA content with that of biosynthesis; and ABA biosynthesis in corn embryos lacking carotenoid precursors with ABA biosynthesis in normal embryos. Embryos were harvested at various times during seed development and divided into two groups. Endogenous levels of ABA were measured in one group of embryos and ABA biosynthetic capacity was measured in the other group. The ABA biosynthetic capacity was measured with and without tetcyclacis (an inhibitor of ABA degradation) in embryos from both field-grown and in-vitro-grown corn kernels. Reduced-carotenoid (either fluridone-treated or genetically viviparous) embryos were also included in the study. Corn kernels developing under field and in-vitro conditions differed from each other in their responses to tetcyclacis and in their profiles of ABA biosynthesis during development. Therefore, in-vitro kernel culture may not be an appropriate substitute for field conditions for studies of embryo development. The developmental profiles of endogenous ABA content differed from those of ABA biosynthesis in isolated embryos of both in-vitro-and field-grown kernels. This indicated that ABA levels in the developing embryos were determined by import from the maternal tissues available to the embryos rather than by in-situ biosynthesis. In embryos with reduced levels of carotenoids, either fluridone-treated or genetically viviparous embryos, ABA biosynthesis was low or nonexistent. This result is expected for the presence of an indirect pathway of ABA biosynthesis and in the absence of ABA precursors.Abbreviations ABA abscisic acid - DAP days after pollination     

葡萄果实中脱落酸结合蛋白的存在及其性质

葡萄果实微粒体上存在高亲和力的脱落酸（ＡＢＡ）结合位点,这些位点与ＡＢＡ的结合具有饱和性,高亲和力及低容量,胰蛋白酶或ＤＴＴ处理可以使该位点的特异结合活性下降约９０％,表明此结合位点是一种蛋白质,故称为ＡＢＡ结合蛋白,它含有维系蛋白质特定构象的二硫键,该蛋白与ＡＢＡ反应的最适ｐＨ为６．０,说明与配基结合部位可能存在带有正电荷的氨基酸残基,结合活性在２５℃高于０℃,结合反应达到动态平衡需要３０ｍｉｎ,３０ｍｉｎ以后结合活性随时间延长而下降。该蛋白与ＡＢＡ结合反应的平衡解离常数为１７．５ｎｍｏｌ／Ｌ,最大结合容量（Ｂｍａｘ）为９８．４ｆｍｏｌ／ｍｇｐｒｏｔｅｉｎ。     

Abscisic acid catabolism enhances dormancy release of grapevine buds

The molecular mechanism regulating dormancy release in grapevine buds is as yet unclear. It was formerly proposed that dormancy is maintained by abscisic acid (ABA)‐mediated repression of bud–meristem activity and that removal of this repression triggers dormancy release. It was also proposed that such removal of repression may be achieved via natural or artificial up‐regulation of VvA8H‐CYP707A4, which encodes ABA 8′‐hydroxylase, and is the most highly expressed paralog in grapevine buds. The current study further examines these assumptions, and its experiments reveal that (a) hypoxia and ethylene, stimuli of bud dormancy release, enhance expression of VvA8H‐CYP707A4 within grape buds, (b) the VvA8H‐CYP707A4 protein accumulates during the natural transition to the dormancy release stage, and (c) transgenic vines overexpressing VvA8H‐CYP707A4 exhibit increased ABA catabolism and significant enhancement of bud break in controlled and natural environments and longer basal summer laterals. The results suggest that VvA8H‐CYP707A4 functions as an ABA degrading enzyme, and are consistent with a model in which the VvA8H‐CYP707A4 level in the bud is up‐regulated by natural and artificial bud break stimuli, which leads to increased ABA degradation capacity, removal of endogenous ABA‐mediated repression, and enhanced regrowth. Interestingly, it also hints at sharing of regulatory steps between latent and lateral bud outgrowth.     

Proteomics in the fruit tree science arena: New insights into fruit defense,development, and ripening

Fruit tree crops are agricultural commodities of high economic importance, while fruits also represent one of the most vital components of the human diet. Therefore, a great effort has been made to understand the molecular mechanisms covering fundamental biological processes in fruit tree physiology and fruit biology. Thanks to the development of cutting‐edge “omics” technologies such as proteomic analysis, scientists now have powerful tools to support traditional fruit tree research. Such proteomic analyses are establishing high‐density 2DE reference maps and peptide mass fingerprint databases that can lead fruit science into a new postgenomic research era. Here, an overview of the application of proteomics in key aspects of fruit tree physiology as well as in fruit biology, including defense responses to abiotic and biotic stress factors, is presented. Α panoramic view of ripening‐related proteins is also discussed, as an example of proteomic application in fruit science.     

A role for jasmonates in climacteric fruit ripening

Jasmonates are a class of oxylipins that induce a wide variety of higher-plant responses. To determine if jasmonates play a role in the regulation of climacteric fruit ripening, the effects of exogenous jasmonates on ethylene biosynthesis and color, as well as the endogenous concentrations of jasmonates were determined during the onset of ripening of apple (Malus domestica Borkh. cv. Golden Delicious) and tomato (Lycopersicon esculentum Mill. cv. Cobra) fruit. Transient (12 h) treatment of pre-climacteric fruit discs with exogenous jasmonates at low concentration (1 or 10 μM) promoted ethylene biosynthesis and color change in a concentration-dependent fashion. Activities of both 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase and ACC synthase were stimulated by jasmonate treatments in this concentration range. The endogenous concentration of jasmonates increased transiently prior to the climacteric increase in ethylene biosynthesis during the onset of ripening of both apple and tomato fruit. The onset of tomato fruit ripening was also preceded by an increase in the percentage of the cis-isomer of jasmonic acid. Inhibition of ethylene action by diazocyclopentadiene negated the jasmonate-induced stimulation of ethylene biosynthesis, indicating jasmonates act at least in part via ethylene action. These results suggest jasmonates may play a role together with ethylene in regulating the early steps of climacteric fruit ripening. Received: 14 August 1997 / Accepted: 4 October 1997     

Abscisic acid influx into human nucleated cells occurs through the anion exchanger AE2

Abscisic acid (ABA) is a hormone conserved from cyanobacteria to higher plants, where it regulates responses to environmental stimuli. ABA also plays a role in mammalian physiology, pointedly in inflammatory responses and in glycemic control. As the animal ABA receptor is on the intracellular side of the plasma membrane, a transporter is required for the hormone’s action. Here we demonstrate that ABA transport in human nucleated cells occurs via the anion exchanger AE2. Together with the recent demonstration that ABA influx into human erythrocytes occurs via Band 3, this result identifies the AE family members as the mammalian ABA transporters.     

Abscisic Acid Content of Algae under Stress*

The distribution of the phytohormone, abscisic acid (ABA), within the phylum of Phycophyta was investigated by an enzyme-linked immunoassay (ELISA). Of 64 algal species tested (originating from 9 divisions, 20 classes and 36 orders, including procaryotes) all species contained ABA, whereas no ABA could be detected in the bacteria Escherichia coli, Rhodospirillum rubrum, and Halobacterium halobium. It is concluded that ABA is universally distributed within the algal kingdom and is not restricted to cormophytes. The ability to synthesize ABA must have been developed even within the procaryotes. The physiological role of ABA in some selected algae was studied by investigating 1. the distribution of ABA between the cells and the culture medium, 2. the responses of endogenous ABA to stress, 3. the synthesis of ¹⁴C-ABA from externally applied ¹⁴C-mevalonic acid, 4. the metabolism of ABA, 5. the effect of externally applied ABA on various physiological reactions of the algae, and the effect of norflurazon on ABA content. ¹⁴C-mevalonic acid served as precursor of ¹⁴C-ABA synthesis in Dunaliella cells and ABA was metabolised to the same products which have been observed in higher plants. In D. parva the internal ABA level increased upon hyperosmotic salt shocks, and in D. acidophila upon alkalization of the medium. Norflurazon caused an increase of ABA content in Dunaliella. Externally applied ABA did not affect photosynthesis, respiration and K⁺ content of the cells. The permeability of the plasma membrane of D. acidophila to water was slightly decreased by ABA. The possible physiological function of ABA in algae is discussed.     

Abscisic acid in developing grains of wheat and barley genotypes differing in grain weight

Two genetically related wheat lines growing in cabinets were given different temperatures during grain filling, and abscisic acid (ABA) was measured in whole grains by gas chromatography with an electron-capture detector. Three genetically related barley lines grown in the field were assayed for ABA content in endosperm and embryo fractions separately by radiommunoassay.Maximum grain growth rate and final weight per grain of the two wheat lines differed by 50–60% at low temperature and 30–40% at high temperature. During grain development two peaks in ABA level were observed at low temperature but only one at high temperature. At times when differences in grain growth rate between genotypes and between temperature treatments were large, the corresponding differences in ABA concentration were small. In barley, one line (Iabo 14) had 30% heavier grains than the other two (Onice and Opale). Endosperm ABA concentrations showed no clear differences between genotypes until grain filling was nearly complete. Embryo ABA levels were up to 10-times greater than those in the endosperm, with Opale having significantly less ABA in the embryo than the other two cultivars.Our experiments did not provide evidence for a causal relationship between ABA levels during grain filling and grain growth rate or final weight.Abbreviations ABA Abscisic acid - DAA days after anthesis - DW dry weight - FW fresh weight     

Abscisic acid relationships in the chill-related dormancy mechanism in apple seeds

Abscisic acid (ABA) levels in seeds from three cultivars of apple (Malus domestica Borkh.) which have substantially different chilling requirements were investigated by gas chromatography mass-spectrometry selected ion monitoring (GCMS-SIM) during stratification. The ABA content of dormant unchilled seeds was similar in the three cultivars, suggesting no relationship between the chilling requirement of those seeds and their ABA status. That chilling is not related to ABA changes during stratification was confirmed by warm (20°C) and cold (5°C) stratification experiments. ABA content dropped rapidly and nearly identically under both temperature regimes, but only cold stratification promoted germination. The decline in ABA during stratification was due in large part to leaching from the seed coat and nucellar membrane; the ABA content of the embryo remained nearly constant. The radicle in intact seeds stratified at 5°C began growing 20–30 days after the ABA in the seed coat and nucellar membrane had nearly disappeared. Radicle growth did not occur in unchilled seeds, even though ABA had leached from them as well. It is possible that the leaching of ABA from the seed allows certain promotive forces to develop, but if so, these can develop only at chilling temperatures. Studies were also conducted on 2-trans ABA relationships to apple seed dormancy, but no association was evident.Report No. 12, Department of Fruit and Vegetable Science, Cornell University.     

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