Interactions between Ethylene, CO(2), and ABA on GA(3)-Induced Amylase Synthesis in Barley Aleurone Tissue

Gibberellic acid-induced synthesis and release of α-amylase in barley aleurone tissue was inhibited by abscisic acid. This inhibition was relieved by simultaneous application of ethylene ranging in concentration from 0.1 to 100 microliters per liter. When CO₂ was applied, it eliminated the effect of 0.1 microliter per liter ethylene and reimposed the abscisic acid inhibition. All concentrations of CO₂ tested from 400 to 10⁵ microliters per liter counteracted the effect of 0.1 microliter per liter ethylene, but had no observable effect on any higher concentration of ethylene. The results indicate that some processes necessary for embryo growth may be subject to regulation by ethylene and carbon dioxide at naturally occurring concentrations of the gases.     

Hypochlorite Disinfection Influences the GA(3)-Induced Synthesis of alpha-Amylase Isozymes by Isolated Barley Aleurone Layers

Aleurone layers isolated from half-seeds of Himalaya barley (Hordeum vulgare cv Himalaya) disinfected in hypochlorite solutions containing 1.0% available chlorine synthesized significantly less α-amylase in response to gibberellic acid than layers derived from half-seeds disinfected in 0.1% hypochlorite. This effect of hypochlorite involved neither a differential decrease in the synthesis of group A or B α-amylase isozymes nor a general decrease in α-amylase synthesis attributable to fewer viable aleurone cells in layers from half-seeds disinfected with 1% hypochlorite. Our results emphasize the need to evaluate the potential effects of routine disinfection procedures used in physiological and biochemical studies.     

光对GA3大麦叶段叶绿素含量的影响

用GA₃处理大麦叶段,在黑暗条件下对叶绿素含量下降无影响,而在光照下有保绿效果。本文就这一现象进行了试验分析,其结果是光的作用在于降低叶内ABA含量,从而减缓叶段内ABA对GA₃的拮抗作用。     

外源GA3和ABA对红肉脐橙果实品质的影响

以生长一致的红肉脐橙成年植株为试材,研究了果实转色前外源GA₃和ABA处理对果实品质的影响。结果表明,外源GA₃处理提高了果皮亮度,但降低了果皮红色度、黄色度和单果重;ABA处理提高了果皮红色度,但降低了果皮亮度;GA₃100mg/L处理虽然极显著降低了果皮厚度,极显著提高了果皮亮度、可溶性固形物和VitC含量,但同时极显著降低了果皮红色度、糖酸比,并极显著提高了果实含酸量;外源ABA处理还极显著降低了果实可食率、出汁率和VitC含量。因此,在果实转色前用外源GA₃和ABA处理红肉脐橙果实均不利于其综合品质的提高。     

外源脱落酸和赤霉素对红肉脐橙果肉糖含量的影响

在红肉脐橙幼果期和着色前分2次喷施不同浓度的外源ABA和GA3,研究其对红肉脐橙果肉糖含量的影响.结果表明:10mg.L-1ABA处理显著或极显著提高了果实成熟时的葡萄糖、果糖和总糖含量,50mg.L-1ABA处理极显著提高了果实蔗糖含量,而100mg.L-1ABA处理极显著降低了果实葡萄糖含量;中低浓度的GA3(10、50和250mg.L-1)极显著提高了果实蔗糖含量,10mg.L-1GA3处理对果实葡萄糖和果糖含量无明显影响,但极显著提高了果实总糖含量,50、250和500mg.L-1GA3处理极显著降低了果实葡萄糖、果糖和总糖含量.表明着色前较低浓度的外源ABA处理(10和50mg.L-1)可提高果实中一种或几种糖的含量,而较高浓度的GA3处理(250和500mg.L-1)则严重阻碍了果肉中糖的积累.     

Observations on the Germ Aleurone of Barley. Morphology and Histochemistry

The germ aleurone over the embryonic axis of barley was examinedin strips of tissue peeled off harvest-ripe grains. The germaleurone is only one cell thick but resembles 'normal' aleuronein being composed of living cells with dense, lipid-rich cytoplasmand thick walls containing phenolic material. In contrast tothe cells of the 'normal' aleurone, germ aleurone cells containvery few phytin or protein deposits. When the germ aleuroneis ruptured during germination, the walls at the torn edge becomethickened with shiny golden-brown material, and 'sealed' tothe testa. Two days after germination, lignin can be detectedin the walls of a single row of germ aleurone cells adjoiningthe scutellum. The role of the germ aleurone in defence againstmicroorganisms is discussed. It is suggested that the metabolicactivities in the germ aleurone in imbibed grains compete withthe embryo for oxygen, and thus may be one of the factors whichdetermine whether a grain germinates or remains dormant.Copyright1994, 1999 Academic Press Barley, Hordeum vulgare L., germ aleurone, histochemistry, defence mechanism, lignin, dormancy, microorganisms, pre-mature germination     

Measurement of the Effects of Acetic Acid and Extracellular pH on Intracellular pH of Nonfermenting, Individual Saccharomyces cerevisiae Cells by Fluorescence Microscopy

The effects of acetic acid and extracellular pH (pH_ex) on the intracellular pH (pH_i) of nonfermenting, individual Saccharomyces cerevisiae cells were studied by using a new experimental setup comprising a fluorescence microscope and a perfusion system. S. cerevisiae cells grown in brewer’s wort to the stationary phase were stained with fluorescein diacetate and transferred to a perfusion chamber. The extracellular concentration of undissociated acetic acid at various pH_ex values was controlled by perfusion with 2 g of total acetic acid per liter at pH_ex 3.5, 4.5, 5.6, and 6.5 through the chamber by using a high-precision pump. The pH_i of individual S. cerevisiae cells during perfusion was measured by fluorescence microscopy and ratio imaging. Potential artifacts, such as fading and efflux of fluorescein, could be neglected within the experimental time used. At pH_ex 6.5, the pH_i of individual S. cerevisiae cells decreased as the extracellular concentration of undissociated acetic acid increased from 0 to 0.035 g/liter, whereas at pH_ex 3.5, 4.5, and 5.6, the pH_i of individual S. cerevisiae cells decreased as the extracellular concentration of undissociated acetic acid increased from 0 to 0.10 g/liter. At concentrations of undissociated acetic acid of more than 0.10 g/liter, the pH_i remained constant. The decreases in pH_i were dependent on the pH_ex; i.e., the decreases in pH_i at pH_ex 5.6 and 6.5 were significantly smaller than the decreases in pH_i at pH_ex 3.5 and 4.5.     

Modulation of Calmodulin mRNA and Protein Levels in Barley Aleurone

Changes in calmodulin (CaM) mRNA and protein were investigated in aleurone layers of barley (Hordeum vulgare L. cv Himalaya) incubated in the presence and absence of calcium, gibberellic acid (GA3), and abscisic acid (ABA). CaM mRNA levels increased rapidly and transiently following incubation of aleurone layers in H2O, CaCl2, or GA3. The increase in CaM mRNA was prevented by ABA. This increase in CaM mRNA was brought about by physical stimulation during removal of the starchy endosperm from the aleurone layer. CaM protein levels did not increase in response to physical stimulation. Only incubation in GA3 plus CaCl2 brought about a rapid increase in CaM protein levels in the aleurone cell. ABA reduced the level of CaM protein below that found at the beginning of the incubation period. The rise in CaM protein preceded increases in the synthesis and secretion of [alpha]-amylase. Immunocytochemistry with monoclonal antibodies to carrot and mung bean CaM was used to localize CaM in aleurone protoplasts. Monoclonal antibodies to tubulin and polyclonal antibodies to tonoplast intrinsic protein and malate synthase were used as controls. CaM was localized to the nucleus, the vacuolar membrane, and the cytosol, but was not associated with microtubules.     

Comparison of IAA-Induced and Low Temperature-Induced GA(3) Responsiveness and alpha-Amylase Production by GA(3) Insensitive Dwarf Wheat Aleurone

Rht3-containing gibberellic acid (GA₃) insensitive deembryonated wheat (Triticum aestivum L. var Cappelle Desprez × Minister Dwarf) aleurone, that can be made responsive to GA₃ by low temperature, can also be rendered GA₃ sensitive by preincubation with indoleacetic acid (IAA). The IAA-induced response of the dwarf selection is concentration-dependent, relatively sensitive, and similar in magnitude to that induced by low temperature. Other auxins also induce GA₃ responsiveness to a greater or lesser degree. IAA has no apparent effect on the wild type (rht, tall) selection.     

Growth-Regulating Substances and the Growth of Tiller Buds in Barley; Effects of IAA and GA3

Application of cytokinin to barley seedlings grown without mineralnutrients leads to rapid growth of coleoptile and first leaftiller buds. IAA and GA₃ cannot substitute for cytokinin inthis effect and applications of TIBA and CCC are also ineffectiveon bud growth. However, when bud growth was promoted, eitherby application of cytokinins or by supplying plants with mineralnutrients, IAA and GA₃ applications caused enhanced tiller growthindicating that these compounds can stimulate growth of activelygrowing buds. The results are compatible with an interpretationwhich stresses the importance of cytokinin availability in determiningtiller bud growth.     

Effects of Cl- on and Malate Fluxes and CO2 Fixation in Carrot and Barley Root Cells

Cl^– and ions interact apparently competitively during influx across the plasmalemma of carrot root cells. Cl^–,however, reduces influx much less than predicted from the effect of on Cl^– influx.Cl^– and plasmalemma influxes both increase with time after excision of carrot tissue. Cl^–and may therefore be transported by a common mechanism. The effect of pH changes on the influx of malate across theplasmalemma in barley roots shows that malate crosses the plasmalemmaas the singly charged anion. Stimulations of influx by bothK₂SO₄ and KCl suggest that the malate anion crosses in associationwith. K⁺. If malate entry is passive, P_mal- is about 2?10^–8cm s^–1, but it is thought that malate entry is partlyan active process. A slight, apparently competitive inhibition by Cl^– ofmalate flux into the vacuole of barley root cells suggests thatthe two anions may be transported by a common process at thetonoplast, but this is not thought to be physiologically significant. The accumulation of ¹⁴C from 1 mM is drastically reduced by 10 mM Cl^–. A quantitative analysis of the kineticsof ¹⁴C exchange shows that Cl^– directly inhibits the formationof malate from . The decreased influx of endogenously produced malate to the vacuole in the presenceof Cl^– is probably a secondary consequence of the fallin the cytoplasmic concentration. The nature of the Cl^–inhibition of malate formation is discussed. In KCl-loaded tissue the influx of external malate and the accumulationof ¹⁴C from external are reduced. The location of these effects is not certain, but the effects suggest thatregulation of malate synthesis and accumulation may be relatedto the negative-feedback regulation of Cl^– and transport.     

Effect of ABA and GA3 on tuberization and some chemical constituents of potato

The effects of abscisic acid (ABA) and gibberellic acid (GA₃)on the chemical composition of and tuberization in the potatowere studied. GA₃ at 10^–6 M inhibited and ABA at an equimolarconcentration promoted tuberization. The inhibitory effect ofGA₃ was relieved by the addition of ABA. The fresh weight ofthe shoot, chlorophyll content and the total glycoalkaloidslevel were increased by GA₃ treatment, but decreased by ABAtreatment. In contrast, total proteins were increased by ABA,but decreased by GA₃ treatment. ¹This work was supported by a PL-480 research grant (FG-Pa-262)from the United States Department of Agriculture widi co-ordinationby the Agriculture Research Council, Pakistan. (Received August 29, 1980; )     

Gibberellin Signaling Pathway:Role of Phospholipids in Barley Aleurone

The present report describes intracellular signaling events elicited by gibberellin (GA) in barley aleurone, where the phytohormone induces an increase on phosphoinositides turnover in a time-dependent manner. A biphasic behavior for PtdInsP was demonstrated by changes in [³²P]PtdInsP levels, the earlier phase peaking at 5 min and the second phase at 25 min while PtdInsP ₂ turnover was rapid but transient, peaking only at 5 min. In contrast, the structural phospholipid PtdCho turnover was only affected after 40 min. In addition, GA stimulated the synthesis of all phospholipids. Incorporation of [₃₂P]Pi in unstimulated aleurone showed that both synthesis and degradation of polyphosphoinositides were relatively faster than those of structural phospholipids. We discuss the possible reasons why the changes in synthesis and turnover of phosphoinositide are required for GA signaling pathway and why it may be a critical event in the control of secretion in aleurone.     

Effect of ABA and GA3 on tuberization and some chemical constituents of potato

The effects of abscisic acid (ABA) and gibberellic acid (GA₃)on the chemical composition of and tuberization in the potatowere studied. GA₃ at 10^–6 M inhibited and ABA at an equimolarconcentration promoted tuberization. The inhibitory effect ofGA₃ was relieved by the addition of ABA. The fresh weight ofthe shoot, chlorophyll content and the total glycoalkaloidslevel were increased by GA₃ treatment, but decreased by ABAtreatment. In contrast, total proteins were increased by ABA,but decreased by GA₃ treatment. ¹This work was supported by a PL-480 research grant (FG-Pa-262)from the United States Department of Agriculture widi co-ordinationby the Agriculture Research Council, Pakistan. (Received August 29, 1980; )     

内源脱落酸和赤霉素对番茄发育果实和种子水分关系的影响

利用热偶湿度计（ｔｈｅｒｍｏｃｏｕｐｌｅｐｓｙｃｈｒｏｍｅｔｅｒ）研究了野生型、ＧＡ－缺陷型和ＡＢＡ－缺陷型番茄发育过程中果实种子的水分关系,发现除ＡＢＡ－缺陷型种子胶囊和果肉水势变化特殊外,３种类型果实水分状况变化基本一致;在整个发育时期内．前期种子胶囊和果肉水分流向种子,中期种子水分流向种子胶囊和果肉,后期种子和果实间的水势达到平衡。鉴于种胚脱水是一种主动过程,种胚水势一直低于整个种子、种子胶囊和果肉。内源赤霉素可明显增加果实和种子的重量,但对增加种胚溶质的作用不大。由于内源脱落酸可以促使果实成熟和衰老,促进果实细胞解体,大大降低种子胶囊和果肉水势,因而抑制成熟种子在果实内萌发。     

Observations on the Germ Aleurone of Barley. Phenol Oxidase and Peroxidase Activity

Strips of tissue containing the germ aleurone layer were removedfrom dry, harvest-ripe grains of barley (Hordeum vulgare L.)and incubated in buffered solutions of phenolic compounds, withand without the addition of hydrogen peroxide. Peroxidase ando-diphenol oxidase activity were found in the material releasedinto the incubation medium, and in the cytoplasm of the germaleurone cells. Peroxidase activity was located in the cellwalls and appeared to be high in the region where the germ aleuronecovering the embryonic axis merges into that which adheres tothe scutellum i.e. the region in which a row of germ aleuronecells becomes lignified following germination. Monophenol oxidaseactivity was not detected in the released enzymes or in theintact tissue. Although hydroquinone was oxidized in the cytoplasmof the germ aleurone tissue, unequivocal evidence of the presenceof laccase was not obtained. The oxidation of endogenous phenolicsubstances by phenol oxidases and peroxidases is discussed inrelation to anti-microbial defence mechanisms which appear tooperate in the germ aleurone during germination.Copyright 1994,1999 Academic Press Barley, Hordeum vulgare L., germ aleurone, catechol oxidase, laccase, peroxidase, defence mechanisms, germination