Effect of cinnamate on nitrate reductase activity in isolated cucumber cotyledons

The effect of cinnamic acid on in vivo nitrate reductase activity and protein content in cucumber cotyledons was studied. Cinnamate increased in vivo nitrate reductase activity and also the total protein content at lower concentrations (0.01–0.1 mM). Higher concentration, however, proved inhibitory. The effect of cinnamate on nitrate reductase activity has been discussed.     

Effect of manganese ions on benzyladenine-induced growth and chlorophyll synthesis in excised cucumber cotyledons

The behaviour of endogenous Mn²⁺ was studied by electron spin resonance spectro-scopy during benzyladenine-induced growth of excised cucumber ( Cucumis sativis L. cv. Long green) cotyledons. The level of endogenous Mn²⁺ was decreased by ben-zyladenine treatment, most pronounced after 96 h. MnCl₂ applied alone promoted chlorophyll synthesis at relatively low concentrations but in the presence of ben-zyladenine higher concentrations of MnCl₂ were required for stimulation of chlorophyll synthesis. A pronounced increase in growth was observed when Mn²⁺ was applied with benzyladenine at 96 h, when the decline in the endogenous level of paramagnetic Mn²⁺ was maximal.     

Phytochrome-mediated development of glycine oxidation by mitochondria in cucumber cotyledons

The development of glycine oxidation activity in mitochondria in etiolated cucumber ( Cucumis sativus L., cv. Shinfushinari) cotyledons is regulated by phytochrome. This conclusion is based on two lines of evidence. 1. The oxidation activity was increased by continuous illumination of far-red light. 2. It was also increased by brief red light pulses, the effect of which was reversed by brief far-red light pulses. The light-induced increase in glycine oxidation and in glycine decarboxylase (EC 2.1.2.10) activity in the cotyledons was inhibited by cycloheximide, but not by chloramphenicol. While glycine oxidation activity continued to increase during light-illumination for 20 h, malate oxidation activity increased for 6 to 8 h after illumination and decreased thereafter. This transient increase in the activity of malate oxidation was also induced by red light pulses and the effect of the red light was reversed by far-red light pulses.     

A double enhancement of greening in the cotyledons of cucumber seedlings by excision and red light

Cotyledons excised without the hypocotyl hook from 6-day-old etiolated cucumber ( Cucumis sativus L. var. Elem) seedlings accumulated a significantly higher amount of chlorophyll than cotyledons excised with hooks or intact cotyledons. It was found that maximum ehancement of greening was achieved after 2 h of dark incubation following excision. Pretreatments with red light effected an additive rise in chlorophyll level in subsequent white light after a dark incubation, suggesting that the effects of excision and phytochrome on greening act independently. Etiolated seedlings were variously dissected before greening and it was found that enhancement occurred only when cotyledons were excised at the level of the hypocotyl hook or above it. Similar results were obtained when the dissected plants were pre-treated with red light.     

三唑酮对黄瓜子叶抗氧化酶活力的影响

黄瓜子叶衰老过程中超氧物歧化酶（ＳＯＤ）、过氧化氢酶（ＣＡＴ）和抗坏血酸－过氧化物酶（ＡＳＡ－ＰＯＤ）活性降低,而过氧化物酶（ＰＯＤ）活性升高。２０ｍｇ／Ｌ三唑酮右明显提高ＳＯＤ,ＡＳＡ－ＰＯＤ,ＣＡＴ活性,抑制ＰＯＤ活性升高。膜脂过氧化产物丙二醛（ＭＤＡ）含量在叶片衰老过程中提高,三唑酮可降低ＭＤＡ含量。表明三唑酮减轻脂氧化程度,延缓了叶片的衰老。     

离体黄瓜子叶不定根发生的研究

离体黄瓜子叶在仅加有双蒸水的培养皿中能高频率发生不定根的现象,并对不定根发生过程中过氧化物酶（ＰＯＤ）活性、可溶性蛋白和核酸含量的变化进行了测定。：６ｄ苗子叶不定根发生率最高,不定根发生高峰是在培养后３ｄ～５ｄ,在不定根发生过程中,过氧化物酶生呈上升趋势,可溶性蛋白和核酸含蜈则降低。     

Nitrate reductase in cotyledons of cucumber seedlings as affected by nitrate, phytochrome and calcium

Regulation by the active form of phytochrome (P_FR) and the effect of Ca²⁺ was examined with nitrate reductase (NR) in etiolated cucumber ( Cucumis sativus cv. Beilpuig). Nitrate reductase activity (NRA) was studied in excised cotyledons of cucumber seedlings grown in distilled water and in darkness for seven days at 24 ± 0.5°C. All experiments were performed in the dark and a dim green safelight was used during analyses. In etiolated cucumber cotyledons NRA was induced by nitrate and a brief irradiation (15 min) with red light (R) resulted in 62% increase in NRA. This effect was nullified when R was followed immediately by a brief (5 min) far-red light (FR). NRA also showed a semidian (12 h) rhythmicity. Both P_FR, and nitrate effects were age dependent. Calcium seemed to be involved since the phytochrome effect was only observed when calcium was supplied in the external solution. The effect of R on NRA depended on the period of calcium nitrate incubation. An external supply of calcium ionophore mimicked the effect of R and, if supplied to R-irradiated cotyledons, produced a higher NR level than that caused by R alone. This suggested that intracellular free calcium was involved.     

Structural-functional changes in detached cucumber leaves, and modelling these by hormone-treated leaf discs

Senescence and rejuvenation were investigated in detached cucumber (Cucumis sativus L.) leaves after cultivation in nutrient solution for one week or four weeks. Rooting of the petiole (visible generally from the 7th day) elicited a combination of different morphological, anatomical, and physiological changes in the lamina. Extensive growth in area and thickness, extreme regreening, changes of chloroplast structure and activity, as well as the pattern of Chl-protein complexes were observed and compared either to the corresponding parameters of young detached leaves or mature attached leaves. These responses could be provoked separately by treating excised leaf discs with kinetin, benzyladenine, or indolylacetic acid. The hormones showed mutuality in their effects, benzyladenine being responsible for the growth of cells, while indolylacetic acid and kinetin promoted an increase in chlorophyll content. However, none of the treatments resulted in the growth of the chloroplasts in the leaf discs, which was only prominent in the rooting leaves.     

三唑酮对离体黄瓜子叶膜系统的影响

研究了三唑酮处理后离体黄瓜子叶衰老过程中活性氧的产生及膜系统有变化。结果表明：20mg/L三唑酮能有效抑制O^-2.、H2O3的累积,减轻质膜相对透性和膜脂过氧化程度及组织自动氧化速率。这表明三唑酮减轻了活性氧在植物体内的累积,可以作为一种活性氧的直接或间接清除剂。     

Benzyladenine induces the appearance of LHCP-mRNA and of the relevant protein in dark-grown excised watermelon cotyledons

Cotyledons were excised from imbibed watermelon seeds, grown for 4 days in darkness on water or 10 M benzyladenine (BA) and then tested for the presence of the light-harvesting chlorophyll a/b protein (LHCP) and its mRNA. LHCP was assayed immunologically by western blotting of SDS gels: the protein was present in plastids, but it was not recovered with the thylakoid fraction. Antibodies directed against LHCP precipitated a 32 kDa polypeptide from translation products of poly(A) RNA of cotyledons only if these had been grown on BA. Taken together the data suggest that in absence of light cytokinins are necessary for the maintenance of a detectable level of LHCP-mRNA as well as for synthesis of the protein.     

Impaired chlorophyll formation in etiolated cucumber cotyledons following prolonged dark incubation after red light pretreatment

Red light (R) pretreatment of etiolated cucumber seedlings ( Cucumis sativus L. var. Elem) followed by prolonged dark incubation prior to white light (WL) exposure, had an adverse effect on the greening of the cotyledons. The effect was photoreversible by far-red (FR) light. Cotyledons which were dark incubated for 24 h following the R pulse greened more rapidly when exposed to WL than did the controls, while total chlorophyll (Chl) accumulation after 24 h in the light was about the same in both. However, after 48 h post-R dark incubation greening of the treated cotyledons was delayed, and their amount of Chl which accumulated after 24 h WL was about one half of that in non-treated seedlings. As the length of the post-R dark incubation period was extended Chl production became slower, so that after 96 h post-R dark incubation the Chl level in the treated cotyledons after 24 h WL was approximately 20% of the controls. No significant differences in amounts of protochlorophyll could be detected between seedlings preilluminated with R or R followed by FR. Seedlings 4-, 5- and 6-days-old at the time of R treatment showed similar degrees of impaired Chl synthesis following prolonged post-R dark incubation.     

Effect of phytochrome activation followed by prolonged dark incubation on chloroplast development in etiolated cucumber cotyledons

The inhibitory effect of a short red light pulse followed by prolonged dark incubation on chlorophyll accumulation in etiolated cucumber cotyledons ( Cucumis sativus L. cv. Elem) was reflected in the development of the internal membrane system of the mesophyll plastids. Dark incubation for 24 h after phytochrome activation produced the characteristic accelerating effect OB chlorophyll synthesis and chloroplast development. However, longer intervening dark periods (48, 72 and 96 h) before white light exposure resulted not only in a diminished capacity to concentrate chlorophyll, but also in an impaired ability to form grana. The absence of stacking was consistent with a high chlorophyll a to chlorophyll b ratio.     

离体培养黄瓜子叶诱导雌花的研究

采用添加Spd和IAA的MS培养基培养离体黄瓜子叶,研究了Spd和IAA对雌花诱导的协同作用,及昼夜温差、培养基中N素和pH值对雌花诱导的影响。结果表明,分别添加Spd、IAA时的雌花诱导率和单株雌花数偏低或为0,12 mg·L^-1 Spd与0.01mg·L^-1 IAA 配合时的诱导效果明显高于单独处理的,而对照组未见雌花,说明Spd和IAA对雌花诱导的协同作用显著。在0、2、6、10℃昼夜温差,60、70、80、90 mmol·L^-1的N素含量和pH 5.4、5.8、6.2、6.6的培养条件下,70 mmol·L^-1 N、6℃温差和pH 6.2时的雌花诱导效果较好,表明适当提高昼夜温差、培养基中N素和pH值有利于黄瓜子叶的雌花诱导。     

Stimulation of chlorophyll synthesis by benzyladenine and potassium in excised and intact cucumber cotyledons

Both benzyladenine (BA) and potassium (K) stimulated chlorophyll synthesis in cucumber ( Cucumus sativus L. cv. National Pickling) cotyledons. However, differences existed between the effects of BA and K. Stimulation of chlorophyll synthesis by BA (1 mg l⁻¹, 4.4 μ M ) was observed in excised cotyledons after 4 and 8 h of illumination but not after 24 h, whereas the stimulation caused by K (40 m M ) continued. In contrast to BA, K was unable to eliminate the lag phase of chlorophyll production, and it also required light for its stimulation of cotyledon expansion. Both BA and K were required to maximize cotyledon expansion and chlorophyll production. In intact plants, K was not limiting for chlorophyll synthesis since foliar or soil pretreatments with K did not markedly stimulate greening. Foliar pretreatment with BA stimulated chlorophyll levels in intact plants, whereas soil pretreatment with BA inhibited chlorophyll production, probably because BA was not readily transported from the roots to the shoot and created a "sink" effect. Inhibitor studies showed that stimulation by K of greening did not depend on RNA or chloroplastic protein synthesis to the extent that has been reported for BA. Thus it appears that BA and K stimulate chlorophyll synthesis via different mechanisms, although both cytokinins and K are essential for maximum rates of greening.     

Phytochrome-mediated control of respiratory activities of mitochondria in cotyledons of dark-grown cucumber seedlings

Mitochondria isolated from cotyledons of dark-grown cucumber ( Cucumber sativus L., cv. Shimotsuki-Aonaga) seedlings after illumination with continuous far-red light showed an increased capacity for oxidation of malate or α-ketoglutarate, as compared with those from cotyledons of non-illuminated seedlings. This increase is supposed to be caused by phytochrome action (high irradiance response). Exogenous NAD⁺ had no effect on the rate of the oxidation of α-ketoglutarate or malate by mitochondria isolated from far-red light-treated cotyledons, but it enhanced the oxidation rate of mitochondria from control cotyledons to the level of mitochondria from light-treated ones. The NAD (NAD⁺+ NADH) content was higher in mitochondria isolated from continuously far-red light-treated cotyledons than in mitochondria from controls. The NAD content was also increased by the treatment with a red light pulse and this response was reversed by a subsequent far-red light pulse. It is proposed that phytochrome controls respiratory activities of cucumber mitochondria by changing the size of the NAD pool in the mitochondria.     

Role of polyamines in adventitious shoot morphogenesis from cotyledons of cucumber <Emphasis Type="Italic">in vitro</Emphasis>

The relationship between polyamines (PAs) metabolism and adventitious shoot morphogenesis from cotyledons of cucumber was investigated in vitro. The endogenous levels of free putrescine (Put) and spermidine (Spd) in the explants decreased sharply, whereas endogenous spermine (Spm) increased during adventitious shoot morphogenesis. The presence of 1–15 mM Put, 1–2 mM Spd, 0.05–1 mM Spm, 5–10 M aminoethoxyvinylglycine (AVG) or 5 M AVG together with 50 M 1-aminocyclopropane-1-carboxylic acid (ACC) in the regeneration medium could promote adventitious shoot formation. Conversely, 1–5 mM D-arginine (D-Arg) or 0.01–0.1 mM methylglyoxal bis-guganylhydrazone (MGBG) inhibited regeneration; and 0.005–0.05 mM ACC displayed little or no evident effects. The explants growing on medium containing 5 M AVG produced higher levels of free Put and Spm, and on medium containing 5 mM Put the explants responded similarly to the AVG-treated explants. However, the exogenous use of 1 mM D-Arg reduced the levels of Put, Spd and Spm, and 0.1 mM MGBG reduced the levels of free Spd and Spm. Moreover, although the explants cultured on medium containing Put and MGBG enhanced ethylene production, AVG and D-Arg inhibited ethylene biosynthesis. This study shows the PAs requirement for the formation of adventitious shoot from cotyledons of cucumber in vitro and the enhanced adventitious shoot morphogenesis may be associated with the elevated level of endogenous free Spm, albeit the promotive effect of PAs on adventitious shoot morphogenesis may not be related to ethylene metabolism.     

Appearance of alternative respiration in cucumber cotyledon mitochondria after treatment with cycloheximide

Treatment of cotyledons of 4-day-old cucumber (Cucumis sativus L. cv. Tokiwa-jibae) seedlings with cycloheximide (0.3 m M ) inhibited protein synthesis in the cotyledons by 80%. In spite of this inhibition, the cycloheximide treatment induced a marked increase in the capacity of the alternative respiration in mitochondria, accompanied by an increase in the contribution of the pathway to the total respiration. In contrast, the activity of the cytochrome pathway was reduced by cycloheximide treatment. As a result, the total respiration was almost the same in mitochondria from cycloheximide-treated cotyledons and untreated cotyledons. Activities of some mitochondrial enzymes examined were also similar. Mitochondrial proteins synthesized during the treatment were separated by two-dimensional gel-electrophoresis and examined by fluorography. No new spots appeared and no spots disappeared on the fluorograms, but the labeling intensity of some polypeptides showed a relative increase or decrease as the result of cycloheximide treatment.     

Uptake and distribution of calcium, magnesium and potassium in cucumber of different age

Uptake and distribution of Ca⁺, Mg²⁺ and K²⁺ were investigated in plants of cucumber ( Cucumis sativus L. var. Cila) which had been cultivated for 12, 19, 32, or 53 days in complete nutrient solution with 1.0 m M Ca²⁺, 2.0 m M Mg²⁺ and 2.0 m M K⁺. The ⁺ concentration was about the same in roots and shoots, while the Ca²⁺ and Mg²⁺ concentrations were low in roots compared to shoots. The K⁺ concentration decreased with increasing leaf age, while the Ca²⁺ and Mg²⁺ concentrations increased, except in older plants with flowers and fruits, where an increased concentration was found in the youngest leaves. This is discussed in connection with increased indoleacetic acid (IAA) synthesis in the shoot. Excision of leaves at different levels from 21-day-old plants, followed by uptake for 24 h from the nutrient solution on days 22 and 23, resulted in no immediate reduction in Ca²⁺ (⁴⁵Ca) uptake. Transport of Ca²⁺ increased to leaves above and below the excision point and total Ca²⁺ uptake remained at the same level as for the intact plant. It is suggested that regulation of Ca²⁺ uptake is primarily achieved in the root while the distribution in the shoot is regulated by the accessability of negative binding sites.     

不同水分处理对日光温室黄瓜多胺与激素的影响

研究了不同土壤含水量对日光温室黄瓜生长与物质代谢的影响 ,结果如下 :冬春茬各生育期随着土壤水分的减少 ,不同处理的叶片数、叶面积、株高、节间长均降低。水分亏缺初期 ,叶片脯氨酸含量随着土壤含水量的降低而增加 ,随着时间延长此趋势消失。盛瓜期 Pro含量以 T1最多 ,达 17.95 μg/ g DW,分别是 T2与 T3的 2 .0倍与 2 .4倍 ;末瓜期各处理的 Pro含量均达最高值 ,T1、T2、T3分别为 2 5 .30、15 .6 0、12 .33μg/ g DW,秋冬茬叶片脯氨酸含量变化规律同冬春茬 .无论冬春茬还是秋冬茬 ,游离蛋白质的变化规律同脯氨酸。叶片亚精胺 (Spd)与腐胺 (Put)含量随着土壤水分的减少而增加。不同水分处理对黄瓜叶片精胺(Spm)含量的影响不大。根系中 Spd、Put、Spm的变化趋势同叶片。冬春茬不同水分处理的黄瓜叶片 ABA含量均随着土壤含水量的减少而增加