Effects of Light on Degradation of Chlorophyll and Proteins during Senescence of Detached Rice Leaves

Regulatory effects of light on senescence of rice leaves wereinvestigated by measuring degradation of chlorophyll and proteinsin leaf segments which had been kept in the dark or under illuminationwith light of different intensities and colors. When leaveshad been left in total darkness for three days at 30°C,there was an initial long lag that lasted for one whole dayand then chlorophyll was rapidly degraded in the second andthird days. Breakdown of chlorophyll was strongly retarded bycontinuous illumination with white light of intensity as lowas 0.5 µmol photons m^–2 s^–1 but the effectof light decreased at intensities above 10 µmol photonsm^–2 s^–2. The initial lag and subsequent degradationof chlorophyll in the dark were little affected by illuminationwith red or far red light at the beginning of dark treatment.However, a brief illumination with red light at the end of thefirst and/or second day significantly suppressed degradationof chlorophyll during subsequent dark periods and the effectof red light was nullified by a short irradiation with far redlight. Thus, degradation of chlorophyll is regulated by phytochrome.Thylakoid membrane proteins and soluble proteins were also largelydegraded during three days in the dark. Degradation of membraneproteins such as the apoproteins of light-harvesting chlorophylla/b proteins of photosystem II and chlorophyll a-binding proteinsof reaction center complexes showed a long lag and was stronglysuppressed by illumination with weak white light. Thus, theloss of chlorophyll can be correlated with degradation of chlorophyll-carryingmembrane proteins. By contrast, light had only a weak protectingeffect on soluble proteins and ribulose-1,5-bisphosphate carboxylase/oxygenaserapidly disappeared under illumination with weak white light.Thus, breakdown of thylakoid membrane and soluble proteins aredifferently regulated by light. Artifacts which would be introducedby detachment of leaves were also discussed. ¹ Present address: Department of Applied Biology, Faculty ofScience and Technology, Science University of Tokyo, Yamazaki,Noda-shi, Chiba, 278 Japan. ² Present address: Department of Life Science, Faculty of Science,Himeji Institute of Technology, Harima Science Park City, Hyogo,678-12 Japan.     

Polyamines Promote the Biosynthesis of Ethylene in Detached Rice Leaves

The effects of polyamines (putrescine, spermidine, spermineand diaminopropane) on the production of ethylene in detachedrice leaves were investigated. Polyamines effectively promotedthe production of ethylene in detached rice leaves under bothlight and dark conditions. Putrescine stimulated the productionof ethylene within 4 hours of its application, a result suggeststhat putrescine enhances the production of ethylene directly.Putrescine also stimulated the production of ethylene in detachedleaves that had been aged for 2 and 4 days. The stimulatoryeffect of putrescine resulted from the enhancement of the synthesisof 1-aminocyclopropane-l-carboxylic acid (ACC) and the conversionof ACC to ethylene. The activity of S-adenosylmethio-nine decarboxylasein segments of rice leaves was inhibited by the applicationof putrescine. Thus, the enhancement of the synthesis of ACCby putrescine seems to be mediated by increases in the activityof ACC synthase and in the level of the substrate (S-adenosylmethionine)for ACC synthase. (Received February 27, 1991; Accepted June 5, 1991)     

Stimulation of Ethylene Production by Exogenous Spermidine in Detached Tobacco Leaves in the Light

Exogenous supply of Spd and Spin stimulated ethylone production in detached tobacco leaves kept in the light. Stimulation, that was first detected after 9 but not 6 h of treatment, linearly increased with concentration and was maintained for several h after returning treated leaves to deionized water. Stimulation of ethylene production was prevented by AVG and Co²⁺ and was accompanied by increased activity of ACC synthase and ethylene-forming enzyme. Put, and other diamines, did not give any stimulatory effect. Stimulation was not accompanied by apparent damage of plasmalomina nor was it reversed by Ca⁺ or Put, suggesting that the cationic properties of polyamines are probably not involved. Stimulation might be due to endogenous polyamine accumulation high enough to inhibit the activity of S-adenosylmethionine decarboxylase, so that all S-adenosyhnethionine might be committed to the ethylene pathway. The stimulatory effect of poly a nines acquires particular interest considering that they have so fur been doscribod to inhibit ethylone production in darkened plant tissues. This finding suggests that polyamines may play a regulatory role in plant development by modulating ethylene bio-synthesis under the control of light.     

Anthocyanin Accumulation Mediated by Blue Light and Cytokinin in Arabidopsis Seedlings

It has been reported that pigmentation In plants Is stimulated by light and cytoklnln （CTK）; however, the signaling pathways and the relationship between light and CTK Involved In the regulation of anthocyanln accumulation remain to be elucidated. We Investigated （i） the role of blue light （BL） and CTK In anthocyanln accumulation ; and （ii） the relationship between BL and CTK In wild type （WT） and by4 mutants of Arabidopsis thaiiana. Two-d-old seedlings grown on medium with or without klnetln （KT） or zeatln （ZT） In darkness were Irradiated using BL at different fluence rates for 3 d before the anthocyanln content was determined using a spectrophotometrlc method. Anthocyanln accumulation was strongly Induced by BL In WT seedlings but not In hy4 seedlings, which demonstrated that CRY1 Is the main photoreceptor for BL. Both KT and ZT enhanced the response of the WT seedlings to BL In a dose-dependent manner, whereas they were not sufficient to promote anthocyanln eccumulatlon In darkness. In addition, data from experiments using the hy4 mutant showed that the CTK effect of BL was also CRYl-dependent. The results from experiments with three different treatment programs showed that the relationship between BL and KT In anthocyanln accumulation of Arabidopsis seedlings seems neither muItlpllcatlve nor additive coactlon, but rather Interaction. BL Is necessary for anthocyanln accumulation, and KT might be Involved In the BL signaling pathway.     

Involvement of Lipid Peroxidation in Water Stress-Promoted Senescence of Detached Rice Leaves

Role of lipid peroxidation and antioxidative enzymes (catalase, peroxidase, superoxide dismutase, ascorbate peroxidase and glutathione reductase) in water stress-promoted senescence of detached rice leaves was investigated. The senescence was followed by measuring the decrease in protein content. Increased lipid peroxidation was closely correlated with senescence in water stressed leaves. Decrease in superoxide dismutase activity was evident 8 h after beginning of water stress. However, decreased catalase, peroxidase, and ascorbate peroxidase activity was observed only when senescence was observed. Glutathione reductase was not affected by water stress. Free radical scavengers retarded water stress-enhanced senescence.     

Ethylene and the Responses to Light of Rice Seedlings

The growth of rice seedlings (Oryza satira L.) in the presence of ethylene caused a change in the response to light of coleoptile elongation. In plants grown in air without added ethylene coleoptile elongation was promoted by red, far-red and yellow-green light only in very young seedlings; in older plants irradiation inhibited the growth of the coleoptile. The effect of growing plants in the presence of ethylene was to prolong the period during which light promoted coleoptile growth. Elongation of the first internode was inhibited by light whether or not the seedlings were grown in the presence of ethylene. A correlation existed between the growth effect of an irradiation and the initial decay rate of phytochrome which was established by the treatment. Regardless of wave length, light sources whose intensities were adjusted to produce a decay rate of about 10% per hour or less induced a moderate rate of coleoptile elongation which persisted for a relatively long period. Irradiation with red or yellow-green light of higher intensity which produced a higher rate of phytochrome decay induced a higher rate of coleoptile elongation, but growth stopped after several hours. Other observations, however, showed that one cannot establish a general simple correlation between the rate of elongation of rice coleoptiles under light and the status of measurable phytochrome in the plant.     

Effects of ABA on Senescence and Cellulase Activity of Detached Rice Leaves

The relationship of cellulase to detached leaf senescence of rice seedlings was investigated by examining the effect of ABA and 6-BA on changes in the level of cellulase of leaf segments during senescence. It was shown that the rise in cellulase activity increased with declining chlorophyll content, which was used as the senescence indicator during the senescence of detached rice leaves caused by ABA. The action of ABA took place only after a 48h lag period. ABA enhanced the cellulase secretion and increased the permeability of plasma membrane. A high level of cellulase activity in cell wall closely related to membrane permeability changes. The action of cellulase in the cell wall may cause depolymerization of β-1, 4-glucan in situ, thus speeding senescence. The 6-BA reverses completely or partly the increase in cellulase activity and tile permeability caused by ABA during the first two day, }) ut it antagonized hardly any of the ABA effect from the third day on, suggesting the onset of an irreversible stage in the senescence of detached rice leaves.     

Effects of Abscisic Acid and Water Stress on the Senescence of Detached Rice Leaves

The effects of abscisic acid (ABA) and water stress on senescence and enzyme activities of oxygen scavenging enzymes of detached rice leaves were compared. Exogenously applied ABA exhibited water stress-like effects by promoting senescence, by decreasing the activities of catalase, peroxidase, ascorbate peroxidase and superoxide dismutase. It seems that the effects of water stress on senescence and enzyme activities are possibly mediated through increased content of endogenous ABA. This revised version was published online in July 2006 with corrections to the Cover Date.     

Senescence of Detached Fern Leaves

Detached fern leaves show a loss in the content of pigments,protein and a decline in 2,6-dichlorophenol indophenol Hillreaction of isolated chloroplasts during aging. Light treatmentof leaves could considerably retard aging induced loss in 2,6-dichlorophenolindophenol photoreduction but it exhibits less effect on pigmentand protein loss. (Received February 7, 1983; Accepted July 14, 1983)     

The Relation between Accumulation of Abscisic Acid and Proline in Detached Rice Leaves

The relation between abscisic acid (ABA) and proline accumulation was investigated in detached rice (Oryza sativa L.) leaves. In darkness, proline content increased about 2-, 2,5- and 6-fold after 24, 48 and 72 h. ABA content reached maximum after 48 h. In the light, proline content remained almost unchanged until 48 h and subsequently increased slightly. ABA content in the light was lower than in darkness, but the maximum was also after 48 h. During 12-h exposure to decreased air humidity, proline content gradually increased, but ABA content increased about 25-fold after 4 h and declined thereafter. Exogenous application of ABA resulted in an increase in proline content in detached rice leaves under both light and darkness.     

Effects of Irradiance and Copper on the Activity of Ascorbate Oxidase in Detached Rice Leaves

The effects of copper on the activity of ascorbic acid oxidasc (AAO) in detached rice leaves under both light and dark conditions and in etiolated rice seedlings were investigated. CuSO₄ increased AAO activity in detached rice leaves in both light and darkness, however, the induction in darkness was higher than in the light. In the absence of CuSO₄, irradiance (40 μmol m^-2 s^-1) resulted in a higher activity of AAO in detached rice leaves than dark treatment. Both CuSO₄ and CuCl₂ increased AAO activity in detached rice leaves, indicating that AAO is activated by Cu. Sulfate salts of Mg, Mn, Zn and Fe were ineffective in activating AAO in detached leaves. CuSO₄ was also observed to increase AAO activity in the roots but not in shoots of etiolated rice seedlings. This revised version was published online in July 2006 with corrections to the Cover Date.     

蓝光对水稻幼苗碳水化合物和蛋白质代谢的调节

分别以白光和黑暗处理为对照,蓝光处理１０ｄ的水稻幼苗碳水化合物含量下降,蛋白质含量增加,亚胺环已酮可抑制蓝光引起的蛋白质含量增加。蓝光处理的水稻幼苗总氨基酸含量比白光和黑暗处理分别提高１２％和３６％,游离态Ｇｌｕ、Ａｓｐ含量也比对照增加。蓝光处理促进水稻幼苗的呼吸作用,对丙酮酸激酶、硝酸还原酶、谷氨酰胺合成酶、谷氨酸合酶和转氨酶的活性表现出一致的促进效应。     

Characteristics of the Induction of the Accumulation of Proline by Abscisic Acid and Isobutyric Acid in Detached Rice Leaves

The effects of abscisic acid and isobutyric acid on levels ofproline in detached rice leaves were compared. The lowest concentrationof abscisic acid that induces accumulation of proline in detachedrice leaves was much lower than that of isobutyric acid. Theaccumulation of porline induced by isobutyric acid was associatedwith acidification of the cell sap, whereas abscisic acid increasedlevels of proline without decreasing the pH of the cell sap.Potassium chloride enhanced the accumulation of proline thatwas induced by abscisic acid, but it did not stimulate thatinduced by isobutyric acid. Of particular interest is the findingthat detached rice leaves treated with fusicoccin showed anincrease in levels of proline and a decrease in the pH of thecell sap. A synergistic increase in levels of proline was observedwhen fusicoccin and abscisic acid applied simultaneously. However,fusicoccin had no effect on the induction of the accumulationof pro-line by isobutyric acid. Benzyladenine inhibited theinduction by abscisic acid of the accumulation of proline, butit did not inhibit the induction by isobutyric acid. It is concludedthat the mode of action of abscisic acid in inducing the accumulationof proline differs from that of isobutyric acid. Our resultsalso indicate that factors other than acidification of the cellsap are involved in regulation of the accumulation of proline. (Received September 25, 1990; Accepted December 20, 1990)     

拟南芥连体和离体叶片光合作用的光响应

测定出 2 5 0 μmol·m-2 ·s-1光强下培养的整株拟南芥连体叶片的光合作用在光强为 80 0 μmol·m-2 ·s-1左右达到光饱和 ,其离体叶片光合作用对光强响应的结果与此类似。自然条件下生长的珊瑚树连体与离体叶片光合作用均在光强约为 10 0 0 μmol·m-2 ·s-1下达到饱和 ,这验证所测得的拟南芥连体和离体叶片光合作用光响应的结果是正确的     

Exogenous Ca2+ Inhibits Senescence-Retarding Effect of Cytokinins in Detached Rice Leaves

Cytokinins and Ca2+ singly retarded senescence of detached rice leaves. When Ca2+ was applied together with cytokinins, the effectiveness of cytokinins was significantly reduced. Ca2+ and cytokinins did not stimulate ethylene production synergistically, ruling out the possibility that ethylene was involved in the inhibition of cytokinin-induced senescence- retarding effect by Ca2+. The experiment with specific compounds known to increase (Ca ionophore A23187), or decrease (EGTA, LaCl3, Verapamil, chlorpromazine) cytosolic Ca2+ level indicated that the elevated cytosolic Ca2+ retards senescence.     

蓝光对水稻幼苗生长及内源激素水平的影响

研究表明：与白光对照相比,蓝光明显抑制水稻幼苗的生长,并使幼苗体内的自由态IAA、GA1、玉米素和二氢玉米素含量下降,ABA含量和乙烯释放量则明显增加。说明蓝光对水稻幼苗生长的影响与其体内激素状况有关。     

Proline, Ornithine, Arginine and Glutamic Acid Contents in Detached Rice Leaves

The effects of water stress on the contents of proline, ornithine, arginine and glutamic acid in detached rice leaves were examined. In water stressed leaves, the content of proline was elevated to a content approximately 8-, 14- and 17-fold higher than in control leaves after 4, 8 and 12 h, respectively. We also observed that omithine and arginine contents were much higher under water stress than in control leaves. However, the content of glutamic acid in water stressed leaves was higher after 4 and 8 h and lower after 12 h than that in control leaves.