多胺与激动素对稀脉浮萍离体叶状体衰老的影响

多胺与ＫＴ 都可抑制暗诱导衰老的稀脉浮萍（Ｌｅｍ ｎａ ａｅｑｕｉｎｏｃｔｉａｌｉｓ）离体叶状体的叶绿素损失,且多胺的作用大于ＫＴ。ＫＴ 还显著抑制蛋白质的损失与蛋白酶活性的上升,而多胺对此却无大的影响。０．０５ ｍ ｍ ｏｌ／Ｌ的甲基乙二醛二脒基－腙（ＭＧＢＧ）轻微促进叶绿素和蛋白质的损失。０．０５ ｍ ｍ ｏｌ／Ｌ的ＫＴ 可抑制衰老过程中腐胺（Ｐｕｔ）的上升和亚精胺（Ｓｐｄ）的下降,而对精胺（Ｓｐｍ ）无明显影响。在稀脉浮萍中,精氨酸脱羧酶（ＡＤＣ）活性占优势。ＫＴ 可轻微促进ＡＤＣ 活性,而对鸟氨酸脱羧酶（ＯＤＣ）和Ｓ－腺苷甲硫氨酸脱羧酶（ＳＡＭＤＣ）活性无显著影响。讨论了多胺与细胞分裂素在抑制植物叶片衰老过程中作用途径的可能关系     

Control of development of amylolytic and proteolytic activities in cotyledons of germinating black gram seeds

The effect of polyamines and related metabolites on several parameters of leaf senescence was followed in detached radish ( Raphanus sativus L. var. radicular cv. "Giant Butter") leaves floated on test solutions in darkness. Leaf senescence was accompanied by a marked loss of chlorophyll, which started at 24–48 h of incubation. The polyamines, spermine and spermidine, and the diamines, putrescine and cadaverine, were highly effective in arresting chlorophyll loss over a period of at least 96 h. l -arginine, and especially l -ornithine, were less active. Polyaminens prevented the marked chlorophyll loss in dark-incubated leaves, but did not compensate for the moderate chlorophyll loss when the leaves were aged in light. Polyamines were also highly effective in retarding earlier events of leaf senescence, prior to chlorophyll loss: both protein degradation and ribonuclease activity were inhibited by spermidine. Chlorophyll and protein loss in dark-or light-incubated suspensions of either "intact" or disrupted chloroplasts was not affected by polyamines. – It is concluded that polyamines are highly effective in preventing chlorophyll loss from detached leaves, possibly by controlling early senescence-linked events which occur in darkness rather than by direct inhibition of chlorophyll degradation.     

The effects of abscisic acid,kinetin and 5-fluorouracil on ribonucleic acid and protein synthesis in senescing radish leaf disks

Summary The effects of abscisic acid and kinetin on RNA synthesis in senescing radish leaf disks were investigated using the improved resolution afforded by polyacrylamide gel electrophoresis. Kinetin stimulated and abscisic acid inhibited incorporation of radioactivity into cytoplasmic ribosomal RNA and soluble RNA. Chloroplast ribosomal RNA synthesis appeared to be confined to the period of leaf expansion and was not detected in fully mature leaves. The effects of kinetin in retarding and of abscisic acid in accelerating leaf senescence were not altered by the inhibition of cytoplasmic ribosomal RNA synthesis with 5-fluorouracil. Following inhibition of cytoplasmic ribosomal RNA synthesis with 5-fluorouracil, kinetin stimulated and abscisic acid inhibited incorporation of radioactivity into polydisperse RNA. These results are discussed in relation to the possible mode of action of kinetin and abscisic acid in senescing leaf tissue.     

Changes occurring during aging and senescence in a submerged aquatic angiosperm (Potamogeton pectinatus)

Changes occurring during aging and senescence of leaves of a submerged aquatic angiosperm ( Potamogeton pectinatus L.) were studied. Total chlorophyll and chlorophylls a and b were maximal in mature, and minimal in old leaves. The chlorophyll a to b ratio was highest in mature leaves. During senescence, the chlorophyll content and the ratio of chlorophyll a to b decreased. The content of DNA, RNA, protein and dry weight, and the activity of alkaline pyrophosphatase decreased while free amino acids, the activity of protease, RNase and acid pyrophosphatase, and the ratio of acid to alkaline pyrophosphatase activity increased during aging and senescence. Kinetin (0.23 m M ) deferred leaf senescence by delaying the loss of chlorophyll, protein, nucleic acids and dry weight, and reducing the rise in free amino acids, the activity of protease, RNase and acid pyrophosphatase and the ratio of acid to alkaline pyrophosphatase activity; while both 0.69 m M ethrel and 0.075 m M ABA hastened senescence. Kinetin pretreatment for an optimum period (12 h) followed by ethrel or ABA treatment partially erased the senescence-promoting effect of the latter. But treatments in a reverse order markedly reduced the delaying effect of kinetin on senescence.     

Changes in chloroplast peroxidase activities in relation to chlorophyll loss in barley leaf segments

Total peroxidase activity increased during senescence of excised barley ( Hordeum vulgare L. cv. Kashimamugi) leaves. Kinetin treatment furter increased total peroxidase activity but repressed chlorophyll degradation in excised barley leaves. When isoperoxidases were extracted from barley leaf segments. 4 cationic and 4 anionic isozymes were found in polyacrylamide gel electrophorests during leaf senescence. The chloroplasts contained only two cationic isoperoxidase activities. One (designated C4) was repressed by kinetin. and the other (C3) was increased by kinetin. Glucosamine, which also repressed the degradation of chlorophyll, completely repressed C4 activity but did not affect C3 activity. The induction with senescence, and the repression with kinetin and glucosamine, suggest chat chloroplast isoperoxidase C4 may function as a chlorophyll-degrading enzyme during barley leaf senescence.     

Interrelationship between ethylene and growth regulators in the senescence of lettuce leaf discs

The interrelationship between ethylene and growth regulators in the senescence of romaine lettuce (Lactuca sativa L.) leaves was studied. Gibberellic acid (GA₃), kinetin, and 3-indoleacetic acid (IAA) retarded chlorophyll loss from leaf discs which were floated on hormone solutions. Abscisic acid (ABA) and ethephon enhanced chlorophyll loss and antagonized the senescence-retarding effect of GA₃ and kinetin. A high concentration of IAA (10^–4 M) caused accelerated chlorophyll loss, whereas a similar concentration of kinetin neither retarded nor promoted chlorophyll loss. The ineffectiveness of IAA and kinetin at their supraoptimal concentrations in retarding leaf senescence was related to increased production of ethylene induced in the treated leaf discs. GA₃ was the most effective in retarding chlorophyll loss and did not stimulate ethylene production at all. The senescence-enhancing effect of ABA was not mediated by ethylene. However, the moderately increased production of ethylene, induced by relatively high concentrations of ABA, could act synergistically with the latter to accelerate chlorophyll loss. It is proposed that the effectiveness of exogenously applied hormones, both in enhancing and retarding senescence, is greatly affected by the endogenous ethylene concentration of the treated plant tissue.Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel, No. 2571-E, 1988 series.     

Effect of the Kinetin-Naphthaleneacetic Acid Interaction Upon Total RNA and Protein in Senescing Detached Leaves

The interaction between kinetin and naphthaleneacetic acid in the regulation of senescence of excised tissue of mature broccoli leaves has been used to examine the extent of synchrony between changes in chlorophyll, RNA, and protein. Kinetin increased the net uptake of (14)C-labeled orotic acid and leucine. Naphthaleneacetic acid decreased the effect of kinetin on net uptake after long treatment, but in short-time treatments the auxin increased the effect of kinetin on net uptake. Results of long (24 hr) treatments indicated a general synchrony between the loss of RNA, protein, and chlorophyll. Naphthaleneacetic acid reduced the stabilizing effect of kinetin upon chlorophyll content and upon the content and synthesis of RNA. In short-time experiments, however, RNA content and synthesis were transiently increased by kinetin, and further increased by kinetin plus naphthaleneacetic acid, while chlorophyll content decreased in the presence of kinetin and decreased further in the presence of kinetin plus naphthaleneacetic acid. Actinomycin-D accelerated the loss of chlorophyll, RNA and protein and strongly depressed the rate of RNA synthesis. In the presence of actinomycin-D the stabilizing effect of kinetin upon RNA was substantially reduced. In contrast, the chlorophyll and protein contents remained higher than in the control. Actinomycin-D did not nullify the basal incorporation of orotic acid into RNA, nor did it negate the effect of kinetin upon incorporation. The failure of synchrony between changes in chlorophyll and RNA does not substantiate the proposal that kinetin regulates senescence by a direct effect upon DNA-dependent RNA synthesis.     

Recovery of loss in chlorophyll and 2,6-dichlorophenol indophenol Hill reaction of isolated chloroplasts during dark induced aging of intact maize seedlings

Loss in the content of pigments and decline in the efficiency of thylakoid membranes to reduce 2,6-dichlorophenol indophenol (DCPIP) have been investigated during dark induced senescence of attached leaves of maize seedlings. The chlorophyll degradation during senescence is differentially inhibited by indoleacetic acid (IAA), gibberellic acid (GA) and kinetin. IAA and GA behave as mild senescence inhibitors in comparison to kinetin. However, in comparison to light, kinetin is relatively less efficient in counteracting senescence. Dark-induced loss in chlorophyll content is fully recovered by light when the dark incubation period is relatively short. The pattern of light recovery of loss in photoelectron transport during dark-aging is similar to the recovery kinetics of chlorophyll. Continuous kinetin treatment of dark-incubated seedlings inhibits the chlorophyll degradation but with decreased duration of kinetin treatment, the efficiency of the hormone to inhibit chlorophyll loss is reduced. The kinetin-induced inhibition of pigment loss is small in comparison with the effect of light.     

Retardation of Leaf Senescence by Ascorbic Acid

Leaf discs of Solatium melongena were floated on various concentrationsof ascorbic acid (AA), gibberellic acid (GA₃), and kinetin inorder to study their effect on senescence. AA was highly effectivein retarding senescence as shown by the arrest of the fall inlevels of chlorophyll, DNA, RNA, and proteins. AA was effectiveat a lower concentration than that of GA₃ or kinetin.     

Production and action of ethylene in senescing leaf discs: effect of indoleacetic Acid, kinetin, silver ion, and carbon dioxide

Supraoptimal concentrations of indoleacetic acid (IAA) stimulated ethylene production, which in turn appeared to oppose the senescence-retarding effect of IAA in tobacco leaf discs. Kinetin acted synergistically with IAA in stimulating ethylene production, but it inhibited senescence. Silver ion and CO(2), which are believed to block ethylene binding to its receptor sites, delayed senescence in terms of chlorophyll loss and stimulated ethylene production. Both effects of Ag(+) were considerably greater than those of CO(2). IAA, kinetin, CO(2), and Ag(+), combined, acted to increase ethylene production further. Although this combination increased ethylene production about 160-fold over that of the control, it inhibited senescence. Treatment with 25 mul/l of ethylene in the presence of IAA enhanced chlorophyll loss in leaf discs and inhibited by about 90% the conversion of l-[3,4-(14)C] methionine to (14)C(2)H(4) suggesting autoinhibition of ethylene production.The results suggest that ethylene biosynthesis in leaves is controlled by hormones, especially auxin, and possibly the rate of ethylene production depends, via a feedback control system, on the rates of ethylene binding at its receptor sites.     

Studies on senescing tobacco leaf disks with special reference to peroxidase

Summary Senescing tobacco leaf disks were treated with hydroxyproline (hypro). Chlorophyll breakdown and -amino-nitrogen increase were partially inhibited, but the decrease in soluble protein was stimulated. The normal increase in absolute acid-phosphatase activity was reversed, but the increase in peroxidase activity occurred although after a lag phase. Kinetin inhibition of senescence was reversed by hypro and kinetin actually reinforced the effects of hypro. Proline only partially overcame the effects of hypro plus kinetin. In the presence of kinetin, proline considerably stimulated the increase in peroxidase activity.Discussed are the way in which hypro may be acting, and the possibility of peroxidase being involved in proline hydroxylation.     

Regulation of arginine decarboxylase by spermine in osmotically-stressed oat leaves

Biosynthesis of polyamines in plants is controlled primarily by the enzymes ornithine decarboxylase (EC 4.1.1.17) and arginine decarboxylase (ADC: EC 4.1.1.19), which are responsible for the production of putrescine, and S -adenosyl-L-methionine (SAM) decarboxylase (EC 4.1.1.50) that is necessary for the formation of spermidine and spermine (Spm). Little is known about the metabolic or molecular mechanisms regulating the synthesis of these enzymes. We have studied the regulation of ADC synthesis by Spm in osmotically-stressed oat ( Avena sativa L. ev. Victory) leaves, using a polyclonal antibody to oat ADC and a cDNA clone encoding oat ADC. Treatment with Spm in combination with osmotic stress resulted in increased steady-state levels of ADC mRNA, yet the levels of ADC activity decreased. This absence of correlation is explained by the fact that Spm inhibits processing of the ADC proenzyme, which results in increased levels of this inactive ADC form and a consequent decrease in the ADC-processed form. Spermine treatment leads to delayed loss of chlorophyll in dark-incubated and osmotically-treated oat leaves. Thus, post-translational regulation of ADC synthesis by Spm may be important in explaining its anti-senescence properties.     

Changes in polyamines and related enzymes with loss of viability in rice seeds

Putrescine, spermidine and spermine of high vigour, low vigour and non-viable (classes 1, 2 and 3 respectively) seeds of Oryza sativa increased with loss of viability. The largest concentration of spermine was found in non-viable embryos. Spermine was absent in the husks of all the three categories of seeds. Arginine decarboxylase was greatest in high vigoured seeds and its activity gradually declined with loss of viability. However, diamine oxidase and polyamine oxidase activities gradually increased with the loss of viability of the seeds while DNA, RNA and protein contents decreased. The total content of polyamines increased on kinetin treatment but declined on ABA treatment. DNA, RNA and protein followed the same trend as polyamines. The polyamine contents increased by ca 3- and 4-fold, respectively, in high vigoured and low vigoured seeds on 10^?4 M kinetin treatment. The activity of ADC followed the same change as that of the polyamines in both cases, but the reverse was observed for the activities of diamine and polyamine oxidases.     

Interaction of naphthaleneacetic Acid and kinetin in the senescence of detached leaves

Kinetin and naphthaleneacetic acid were applied separately and in combination to excised leaf disks, localized areas of laminae, and petioles of detached leaves of broccoli (Brassica oleracea L., var. italica) and Xanthium (Xanthium pensylvanicum Wallr.). Senscence (measured as loss of chlorophyll) was strongly retarded by kinetin, but very slightly influenced by naphthaleneacetic acid. When the 2 substances were applied concurrently, the effect of kinetin was markedly reduced by naphthalene acetic acid. Neither interference with uptake nor transport of kinetin appeared to cause the reduction.     

Localized Effect of Polyamines on Chlorophyll Loss

The effect of polyamines, spermidine and spermine, on senescenceof detached leaves of rice, wheat and soybean was investigated.The decline of chlorophyll in rice leaf segments was promotedby polyamines during the first three days and in soybean leafdiscs during the first day of incubation. However, the declinein chlorophyll was later retarded by polyamines. Polyamineswere more effective in retarding senescence of detached wheatleaves at a higher concentration (10 mM) or in narrower segments.The retardation of chlorophyll loss by polyamines was mainlylocalized in those areas around the cut edges of detached leavesor near large veins of soybean primary leaves. This suggeststhat polyamines are not readily transported in leaf cells. ¹ Supported by research grant to C. H. K. from the NationalScience Council of the Republic of China (NSC 72-0409-B002-18). (Received June 4, 1983; Accepted September 12, 1983)     

Control of senescence in rumex leaf discs by gibberellic Acid

The kinetics of chlorophyll and protein decomposition and the effect of gibberellic acid (GA) were examined in senescing leaf discs of Rumex crispus and R. obtusifolius. Loss of Rumex total chlorophyll proceeds at a slow rate for about 2 days followed by a period of rapid logarithmic decline. Chlorophyll b is lost at a slightly faster rate than chlorophyll a during senescence in discs as well as in situ. GA causes a complete cessation of net chlorophyll and protein degradation for several days in Rumex, in contrast to the incomplete senescence inhibition generally observed with cytokinins. GA is fully effective even when added at the middle of the logarithmic phase of chlorophyll loss. Senescence inhibition by GA is apparently gradually reversed upon GA removal. The cytokinins, kinetin and 6-benzylaminopurine, were also effective in Rumex leaf discs, indicating that the senescence retarding effect was not restricted to the gibberellins.     

L-精氨酸与精胺延缓大麦离体叶片衰老效应的比较

L-精氨酸和精胺均具有延缓大麦离体叶片叶绿素,蛋白质和核酸含量下降的作用。L-精氨酸具有良好的渗透性和移动性。但其最适浓度高于精胺100倍,作用时间也滞后于精胺。L-精氨酸可使叶片内多胺含量增加,多胺合成抑制剂ABA则削弱L-精氨酸延缓叶片衰老的作用。L-精氨酸转变成具有强烈生理活性的多胺可能是其作用机理之一。     

The Changes in Soluble Protein of Excised Barley Leaves during Senescence and Kinetin Treatment

Sterile detached barley leaves were floated on water or kinetin (10 mg/1)and supernatant extracts (30,000 x g for 30 min) were prepared from these leaves over an 8 day incubation period. Changes in selected total enzyme levels and in individual soluble protein components wore compared. Ribonuclease, deoxyribonuclease and peptidase activities rose in senescing leaves, even as total protein levels fell. Kinetin to some extent depressed these activities. Evidence of considerable loss of ribonuclease and to a lesser extent of deoxyribonuclease into the surrounding medium was obtained. Soluble supernatant ant extracts were resolved on DEAE-Sephadex (A-50) columns into about 15 components. While most components were degraded during senescence they did so at different rates. Kinetin lowered the rate of degradation of all components. Since no conclusive evidence of a new protein(s) was obtained in water and kinetin treated leaves, it was considered that any protein changes may have been of a quantitative rather than qualitative nature.     

L-精氨酸与精胺延缓大麦离体叶片衰老效应的比较

L-精氨酸和精胺均具有延缓大麦离体叶片叶绿素、蛋白质和核酸含量下降的作用。L-精氨酸具有良好的渗透性和移动性。但其最适浓度高于精胺100倍,作用时间也滞后于精胺。L-精氨酸可使叶片内多胺含量增加,多胺合成抑制剂ABA则削弱L-精氨酸延缓叶片衰老的作用。L-精氨酸转变成具有强烈生理活性的多胺可能是其作用机理之一。