脱落酸及其类似物对玉米线粒体异柠檬酸脱氢酶活性的影响

用（＋）ＡＢＡ及其两种ＲＣＡ系列类似物（ＲＣＡ．７ａ,ＲＣＡ．７ｂ）处理主米黄化芽提取的离体线粒体,三者均能促进线粒体上异柠檬酸脱氢酶（ＩＣＤＨ）的活性,另外,（＋）ＡＢＡ、ＲＣＡ．７Ａ及ＲＣＡ．７ｂ分子都有一个环己烯酮环（ｃｙｃｌｏｈｅｘｅｎｏｎｅ ｒｉｎｇ）,差别仅在侧链的不同,这提示,此环己烯酮环是ＡＢＡ表现此种促进作用所必需的。（＋）ＡＢＡ处理离体线粒体之前,先经抗ＡＢＡ结合蛋白的抗体（ａ     

一氧化氮是脱落酸诱导杨树叶片气孔关闭的信号分子

研究了外源NO和ABA对杨树气孔运动调节作用．结果表明,外源NO和ABA都能诱导杨树离体叶片气孔关闭,且具有剂量效应,NO可加强ABA诱导气孔关闭的作用．NO清除剂(c—PTIO)可大大减弱NO和ABA对气孔关闭的诱导作用．证实了NO参与ABA调控气孔开闭运动过程,不同浓度NO供体SNP和ABA处理杨树离体叶片,SOD活性变化不明显,POD活性受到显著抑制．杨树叶片粗酶液的体外实验表明,不同浓度SNP对POD活性的抑制呈明显的浓度及时间效应;而ABA对POD活性则几乎没有影响．本研究证明,NO调节ABA诱导的树木气孔关闭作用,是ABA诱导树木气孔关闭的一种重要信号分子．     

ABA和NaCl对碱蓬多胺和脯氨酸代谢的影响

比较了 5 0 μmol/L的ABA和 4 0 0mmol/L的NaCl对碱蓬植株内脯氨酸 (Pro)和多胺 (PAs)含量和代谢关键酶活性的影响。结果表明 :ABA处理能诱导碱蓬植株吡咯啉 5 羧酸合成酶 (P5CS)、鸟氨酸脱羧酶(ODC)、多胺氧化酶 (PAO)和脯氨酸脱氢酶 (ProDH)活性上升 ,Pro和游离多胺含量增加。ABA NaCl处理促进碱蓬植株体内Pro和PA的积累。与NaCl处理相比 ,ABA NaCl处理后碱蓬植株P5CS活性显著上升 ,ProDH活性变化不明显 ,ODC活性极显著上升 ,PAO和转谷酰胺酶 (TGase)活性上升 ,从而使Pro和游离多胺含量均显著上升 ,游离态多胺 (Spd PAx) /Put提高 ,结合态多胺总量上升。ABA NaCl处理对碱蓬植株体内Pro和PAs合成的促进与其对生长速率的促进效应相一致     

茉莉酸甲酯和脱落酸对绿豆下胚轴质膜H+-ATPase水解活性的影响

茉莉酸甲酯(MeJA)促进绿豆下胚轴质膜H+-ATPase水解活性.活体条件下,50μmol·L-1 MeJA处理7 h的酶活性提高30%;离体条件下,10 μmol·L-1 MeJA处理2 h的酶活性最大,即提高30%.壳梭孢素(FC)和MeJA在离体条件下对H+-ATPase活性的促进效应相同,均提高30%左右,无协同效应;活体条件下,FC促进质膜H+-ATPase水解活性可达70%,而MeJA仅为30%.离体条件下,脱落酸(ABA)对H+-ATPase水解活性无明显促进;而活体条件下则有一定的抑制.     

脱落酸对甜椒幼苗抗寒性的诱导效应及其机理研究

以砂培甜椒幼苗为材料,先用不同浓度脱落酸(ABA)进行灌根处理,再分两组分别于常温(25℃～30℃)生长7 d(常温苗)或低温驯化(10℃～15℃)30 d(驯化苗)之后进行低温胁迫(5℃)处理,分析幼苗的生长状况及其细胞质膜伤害、渗透调节物质、活性氧清除系统、内源激素的变化特征,以探讨脱落酸对甜椒抗寒性的诱导效应及机理.结果显示,(1)各浓度ABA对甜椒常温苗和驯化苗的株高、茎重均无显著影响,但显著促进驯化苗的侧根生长,在低温胁迫下,显著降低驯化苗叶片的呼吸速率;(2)常温苗的相对电导率仅在10 mg/L ABA处理中比对照显著降低,而驯化苗在各处理浓度下均显著降低,MDA含量表现出与此类似的变化;(3)叶片脯氨酸、可溶性糖、可溶性蛋白、钾离子等渗透调节物质含量在各浓度ABA处理常温苗和驯化苗中均有所增加,但仅脯氨酸含量的增加达到显著水平.(4)各浓度ABA处理都一定程度上提高了常温苗的SOD活性,显著降低了常温苗的CAT和POD活性,H2O2积累量均显著降低,但对于驯化苗,1.0 mg/L和10 mg/L ABA处理却显著降低其SOD活性,POD和CAT活性无显著差异,H2O2积累量也无显著变化.(5)茎尖的ZR、JA-Me含量及IAA/ZR、均与对照无显著差异,但IAA、ABA含量、ABA/ZR比值明显提高,其中1.0和10 mg/L处理的内源ABA含量以及10 mg/L处理的ABA/ZR比值显著高于对照.研究表明,脱落酸能通过降低甜椒幼苗呼吸速率,提高叶片脯氨酸、可溶性糖、钾离子等渗透调节物质以及ABA的积累来诱导增强甜椒幼苗的抗寒性,减少活性氧自由基的产生和累积量,从而减轻低温胁迫造成的伤害,且低温驯化条件下比常温下效果更加明显.     

脱落酸对玉米胚芽和大豆子叶线粒体呼吸的影响

在提取玉米和大豆线粒体后,以ABA处理线粒体,发现在不同底物存在下,ABA均可增加线粒体的耗氧速率,呼吸速率随ABA浓度变化的结合常数（Kd）值为1．43μmol／L。ABA对4态呼吸的促进作用更为显著,因而导致呼吸控制下降,P／O比降低。蛋白质合成抑制剂环己酰亚胺不影响ABA的效应。以ABA预处理大豆子叶,虽然也促进了呼吸作用,但不改变呼吸控制和P／O.     

低温胁迫下脱落酸对线粒体膜磷脂酶D活性的影响

高山离子芥(Choraspora bungeana)是一种稀有高山冰缘植物,其生活环境具有低温、强紫外线等胁迫因子。PLD在膜磷脂降解及磷脂信号转导过程中发挥着重要作用,但其活性往往受到多种因素的影响。该研究以高山离子芥试管苗为材料,研究了4℃、0℃和-4℃胁迫下,ABA对高山离子芥试管苗叶中线粒体膜结合态PLD活性的影响。结果表明:10,50和100μmol·L~(-1)脱落酸(ABA)处理高山离子芥后,线粒体膜结合态PLD活性均较未添加ABA的处理组线粒体膜结合态PLD活性高,其中以50μmol·L~(-1) ABA对离子芥叶中线粒体膜结合态PLD活性的促进作用最为显著;外施0.3 mmol·L~(-1)的ABA合成抑制剂钨酸钠处理高山离子芥后,线粒体膜结合态PLD活性较对照组线粒体膜结合态PLD活性降低;在50μmol·L~(-1) ABA+5 mmol·L~(-1) EGTA处理组中,高山离子芥叶中线粒体膜结合态PLD活性低于未添加EGTA处理组线粒体膜结合态PLD活性;在0.3 mmol·L~(-1)钨酸钠+10 mmol·L~(-1)CaCl_2处理组中,高山离子芥叶中线粒体膜结合态PLD活性高于未添加CaCl_2处理组线粒体膜结合态PLD活性。由此推测,低温胁迫下ABA可能通过Ca~(2+)介导影响高山离子芥叶中线粒体膜结合态PLD的活性。     

ABA信号转运调节的基因表达与源库动力学分析

通过对拟南芥NCED3、AA03及SDR1蛋白亚细胞定位分析及根系和叶片ABA池的动态库变化研究,结果表明气孔运动的有效ABA信号来自于保卫细胞之外,SDR与ABA前体加工和运输有关。胁迫处理后根系合成酶基因转录水平显著高于叶片,但叶片ABA水平是根系的10倍以上,离体叶片和附体叶片ABA含量测定表明,叶片ABA池的形成主要决定于根源ABA的输入。氟啶酮药剂阻断和遮荫实验说明根系ABA池受叶源类胡萝素前体供应影响。叶片ABA水平受根源ABA和叶源类胡萝素前体库双向转运调节,维管束组织系统可能协同和整合了这一复杂调节机制。该结论为逆境ABA信号转递机制研究和操纵内源ABA含量增强植物抗逆性的应用提供相关资料。     

玉米根质膜ABA结合蛋白的增溶及特性研究

以玉米(Zea mays L.)根为材料,采用分级离心和两相分配法制备高纯度的质膜。实验比较了Tri-ton X-100和丙酮两种增溶剂对膜上ABA结合蛋白(ABA-BPm)的增溶效果。结果表明0.2％(W/V) TritonX-100的增溶效率超过85％,而丙酮法增溶效率仅达65％。放射配基(~3H-ABA)结合分析表明,增溶的膜蛋白与ABA的结合反应具有竞争性、饱和性、专一性和高亲和性等特点,而相同反应条件下的BSA则没有这些特征,证明了质膜上存在着ABA结合蛋白。实验还比较了ABA-BPm与增溶的ABA结合蛋白(ABA-BPs)与ABA的特异结合活性,结果显示ABA-BPs对反应温度和介质pH更为敏感,保持最大结合活性的时间也较短(<30min),暗示着增溶膜蛋白离开膜脂环境后更不稳定、易失活。     

水稻幼叶中与ABA亲和力强的结合蛋白

在水稻幼苗叶片中存在膜结合的 ABA 专一结合位点。专一结合位点对 ABA 具有强的亲和力,其与 ABA 反应的平衡解离常数为2.69×10~(-7)mol/L,总浓度为4 nmol mg~(-1)蛋白质。专一结合位点与 ABA 结合活力在0℃时比25℃时高115%。专一结合位点与 ABA 结合的最适 pH 为4.5。ABA 与其专一结合位点的结合量随反应时间延长而增加,1小时达最大值,以后又逐渐降低。这种降低可能是由于专一结合使点活性逐渐减弱所致。这类结合位点可能是 ABA 进入细胞的载体,也可能是 ABA 作用的受体。     

用ANS研究脱落酸提高植物线粒体膜的流动性

用荧光标记法,以ＡＮＳ作荧光探剂研究了脱落酸（ＡＢＡ）对植被线粒体脱流动性的影响。（＋）ＡＢＡ浓度效应实验结果和不同ＰＨ及不同温度条件下的实验结果均表明（＋）ＡＢＡ能降低植物线粒体膜的荧光强度,即（＋）ＡＢＡ能提高线粒体原流动性,且此种效应具有浓度饱和性,并导致线粒体异柠檬酸脱氢酶（ＩＣＨ）活性升高。ＡＢＡ的类似物,ＲＣＡ．７Ａ和ＲＣＡ．７ｂ,也具有提高ＩＣＤＨ活性的效应,抗ＡＢＡ结合蛋白的抗体（     

Effect of the minor ABA metabolite 7'-hydroxy-ABA on Arabidopsis ABA 8'-hydroxylase CYP707A3

To examine the effect of the minor abscisic acid (ABA) metabolite 7'-hydroxy-ABA on Arabidopsis ABA 8'-hydroxylase (CYP707A3), we developed a novel and facile, four-step synthesis of 7'-hydroxy-ABA from alpha-ionone. Structural analogues of 7'-hydroxy-ABA, 1'-deoxy-7'-hydroxy-ABA, and 7'-oxo-ABA were also synthesized to evaluate the role of the 7'-hydroxyl group on binding to the enzyme. The result of enzyme inhibition assay suggests that the local polarity at C-7', neither steric bulkiness nor overall molecular hydrophilicity, would be the major reason why (+)-7'-hydroxy-ABA is not a potent inhibitor of CYP707A3.     

An enzyme-immunoassay of abscisic acid in potato (Solanum commersonii) cultured cells

Estimation of abscisic acid (ABA) content in potato (Solanum commersonii) suspension-cultured cells with an enzyme-immunoassay (EIA) was investigated. In crude extracts of potato cultured cells or even after simple clean-up using C₁₈ cartridge, EIA based on commercial monoclonal antibodies (Idetek, Inc) failed to detect any ABA content. An interference could be removed by partitioning against ethyl acetate after the C₁₈ cartridge so that the EIA yielded an estimate of ABA similar to that determined by high pressure liquid chromatography and gas chromatography-mass spectrometry analysis. These results demonstrate the presence of metabolites in potato cultured cell extract that prevent the binding of ABA to its binding site but not the binding of tracer.Abbreviations ABA abscisic acid - EIA enzyme-immunoassay - HPLC high performance liquid chromatography - GC-MS gas chromatography-mass spectrometry     

Synthesis and biological activity of 4'-methoxy derivatives of abscisic acid

Replacing the 4'-carbonyl group of abscisic acid with a methoxy group does not affect the abscisic acid (ABA)-like activities of the product in barley aleurone protoplasts, although the reduction of ABA to 4'-hydroxyl derivatives significantly reduces the ABA-like activity of the products. This suggests that methoxy derivatives of abscisic acid might be used to produce probes for ABA binding proteins.     

Effects of Abscisic Acid Metabolites and Analogs on Freezing Tolerance and Gene Expression in Bromegrass (Bromus inermis Leyss) Cell Cultures

Optical isomers and racemic mixtures of abscisic acid (ABA) and the ABA metabolites abscisyl alcohol (ABA alc), abscisyl aldehyde (ABA ald), phaseic acid (PA), and 7[prime]hydroxyABA (7[prime]OHABA) were studied to determine their effects on freezing tolerance and gene expression in bromegrass (Bromus inermis Leyss) cell-suspension cultures. A dihydroABA analog (DHABA) series that cannot be converted to PA was also investigated. Racemic ABA, (+)-ABA, ([plus or minus])-DHABA, and (+)-DHABA were the most active in inducing freezing tolerance, (-)-ABA, ([plus or minus])-7[prime]OHBA, (-)-DHABA, ([plus or minus])-ABA ald, and ([plus or minus])-ABA alc had a moderate effect, and PA was inactive. If the relative cellular water content decreased below 82%, dehydrin gene expression increased. Except for (-)-ABA, increased expression of dehydrin genes and increased accumulation of responsive to ABA (RAB) proteins were linked to increased levels of frost tolerance. PA had no effect on the induction of RAB proteins; however, ([plus or minus])- and (+)-DHABA were both active, which suggests that PA is not involved in freezing tolerance. Both (+)-ABA and (-)-ABA induced dehydrin genes and the accumulation of RAB proteins to similar levels, but (-)-ABA was less effective than (+)-ABA at increasing freezing tolerance. The (-)-DHABA analog was inactive, implying that the ring double bond is necessary in the (-) isomers for activating an ABA response.     

The Effect of Abscisic Acid on the Metabolism of Kinetin in Detached Leaves of Rumex pulcher

The effect of abscisic acid (ABA) and of gibberellic acid (GA₂)on the metabolism of kinetin in detached leaves was investigated.Kinetin labelled in either the purine ring or the N-6 substitutedside chain was fed into leaves of Rumex pulcher. The leaveswere extracted 4 or 20 h later and the extract chromatographedon paper which was subsequently scanned for radioactivity. Fourmajor radioactive peaks were revealed. Two peaks were tentativelyidentified, one kinetin and its riboside, and the other adenine.ABA effected a significant decrease in the transformation ofkinetin to adenine while GA₃ had no consistent effect. The possiblemeaning of this finding is discussed.     

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     

Abscisic acid regulation of heterophylly in Marsilea quadrifolia L.: effects of R-(-) and S-(+) isomers

The plant hormone abscisic acid (ABA) induces a developmental switch in the aquatic fern Marsilea quadrifolia, causing the formation of aerial type characteristics, including the elongation of petioles and roots, a change in leaf morphology, the expansion of leaf surface area, and the shortening of the internodes. A number of ABA-responsive heterophylly (ABRH) genes are induced early during the transition. Using optically pure isomers of ABA, it was found that both the natural S-(+)-ABA and the unnatural R-(-)-ABA are capable of inducing a heterophyllous switch and regulating ABRH gene expression. When dose responses are compared, the unnatural ABA gives stronger morphogenic effects than the natural ABA at the same concentration, it is effective at lower concentrations, and its optimal concentration is also lower compared with the natural ABA. Deuterium-labelled ABA enantiomers were used to trace the fate of applied ABA and to distinguish the applied compound and its metabolites from the endogenous counterparts. In tissues, the supplied (+)-ABA was metabolized principally to dihydrophaseic acid, while the supplied (-)-ABA was converted at a slower rate to 7'-hydroxy abscisic acid. Treatment with either enantiomer resulted in increased biosynthesis of ABA, as reflected in the accumulation of endogenous dihydrophaseic acid. Taken together, these results suggest two distinct mechanisms of action for (-)-ABA: either (-)-ABA is intrinsically active, or its activity is due to the stimulation of ABA biosynthesis.     

RopGEF2 is involved in ABA‐suppression of seed germination and post‐germination growth of Arabidopsis

The involvement of Rho of Plants (ROP) GTPases in abscisic acid (ABA) signalling in Arabidopsis has been demonstrated in many studies. However, the roles of RopGEFs (Rop guanine nucleotide exchange factors), which modulate ROP activities in ABA signalling, are poorly understood. Here, we demonstrate that RopGEF2 may play a negative role in ABA‐suppressed seed germination and post‐germination growth. We show that disruption of RopGEF2 enhances sensitivity to exogenous ABA in seed germination assays and that RopGEF2pro‐GUS is mainly expressed in developing embryos and germinating seeds. Interestingly, YFP‐RopGEF2 is located in both the cytoplasmic region and in mitochondria. Notably, the PRONE2 (plant‐specific ROP nucleotide exchanger 2) domain of RopGEF2 is detected in mitochondria, whereas the N‐terminus of RopGEF2 is shown to be in the cytosol. After ABA treatment, degradation of RopGEF2 is triggered in the cytosol through the ubiquitin‐26S proteasome system. The binding of RopGEF2 to ROP2, ROP6 or ROP10, which has been demonstrated to be involved in ABA signalling, not only alters the localization of RopGEF2 but also enables RopGEF2 to escape degradation in the cell. Thus, in this study, we deduce a sophisticated mechanism of ABA‐mediated RopGEF2‐ROP signalling, which potentially implicates the inactivation of ROPs in responsiveness to ABA.     

Interrelations between Gibberellic Acid, Cytokinins and Abscisic Acid in Retarding Leaf Senescence

The interrelation between the effects of abscisic acid (ABA) and the effects of cytokinins and gibberellic acid in retarding leaf senescence was investigated. Leaf discs from plants of Taraxacum megallorrhizon, Rumex pulcber and Tropaeolum majus were floated on solutions of cytokinin or GA to which given amounts of ABA were added. After five days, chlorophyll was extracted and the amount estimated spectrophoto-metrically. The interrelation between the effects of abscisic acid and cytokinins differed from that between the effects of ABA and gibberellic acid. Abscisic acid reduced the senescence retarding effect of GA more than that of cytokinins. A high concentration of cytokinins nullified the senescence enhancing effect of low concentrations of ABA. GA did not reverse the effects of ABA.