Senescence of rice leaves XXXI. Changes of chlorophyll,protein, and polyamine contents and ethylene production during senescence of a chlorophyll-deficient mutant

Leaf senescence of a chlorophylldeficient rice mutant (LT-8) was investigated. At 10 days after planting, the chlorophyll level in the third leaves of rice seedlings of the mutant was about one half that of normal leaves (Norin no. 8), whereas no difference in the protein level could be detected in the two genotypes. The protein level in leaves decreased with increasing age, and no significant difference could be detected during senescence in the two genotypes. Chlorophyll level in the normal leaves also decreased with increasing age. However, the chlorophyll level in the mutant leaves began to decrease only after more than 60% of the initial protein had been degraded. The pattern of ethylene production in the normal leaves was, in general, similar to that in the mutant leaves. Ethylene production first decreased with age, increased to a maximum at day 18, and decreased thereafter. Both spermidine and spermine levels in the leaves of the two genotypes decreased with increasing age. The pattern of the putrescine level in the normal leaves behaved somewhat similar to that in the mutant leaves. However, during the course of senescence, the putrescine level in the mutant leaves was always higher than that in the normal leaves. The possible relationship between endogenous polyamine levels and ethylene production is discussed.     

Senescence of rice leaves XXXI. Changes of chlorophyll, protein, and polyamine contents and ethylene production during senescence of a chlorophyll-deficient mutant

Leaf senescence of a chlorophylldeficient rice mutant (LT-8) was investigated. At 10 days after planting, the chlorophyll level in the third leaves of rice seedlings of the mutant was about one half that of normal leaves (Norin no. 8), whereas no difference in the protein level could be detected in the two genotypes. The protein level in leaves decreased with increasing age, and no significant difference could be detected during senescence in the two genotypes. Chlorophyll level in the normal leaves also decreased with increasing age. However, the chlorophyll level in the mutant leaves began to decrease only after more than 60% of the initial protein had been degraded. The pattern of ethylene production in the normal leaves was, in general, similar to that in the mutant leaves. Ethylene production first decreased with age, increased to a maximum at day 18, and decreased thereafter. Both spermidine and spermine levels in the leaves of the two genotypes decreased with increasing age. The pattern of the putrescine level in the normal leaves behaved somewhat similar to that in the mutant leaves. However, during the course of senescence, the putrescine level in the mutant leaves was always higher than that in the normal leaves. The possible relationship between endogenous polyamine levels and ethylene production is discussed.     

Relation of polyamine synthesis and titer to aging and senescence in oat leaves

Polyamine biosynthesis in senescing leaves of Avena sativa L. was measured by determining the activities of arginine decarboxylase (EC 4.1.1.19), ornithine decarboxylase (EC 4.1.1.17) and S-adenosyl-l-methionine decarboxylase (EC 4.1.1.50). Polyamine content was also estimated by thin layer chromatography and high performance liquid chromatography. Arginine decarboxylase activity decreases progressively in aging attached first leaves and in senescing excised leaves in the dark. Conversely, it increases during light exposure of excised leaves, which retards senescence. Ornithine decarboxylase activity is high and constant in the attached leaf, irrespective of age; it decreases in excised leaves kept in the dark and in the light, irrespective of senescence. S-Adenosyl-l-methionine decarboxylase shows no correlation with age or senescence. Levels of putrescine, diaminopropane, agmatine, and spermidine are high in young leaves and decline with age. The best single indicator of senescence is usually spermidine, which decreases in excised leaves incubated in the dark, but increases in such leaves with time of light exposure. Spermidine generally has a reciprocal relationship with putrescine, indicating that spermidine synthase, which converts putrescine to spermidine, may exert important physiological control. These data support the view that polyamines play an important role in the regulation of plant development.     

Endogenous gibberellin and abscisic Acid content as related to senescence of detached lettuce leaves

Levels of gibberillins (GAs) and of abscisic acid (ABA) in attached leaves of romaine lettuce (Lactuca sativa L.) declined as the leaf became older. The time course of changes in hormone levels, determined in detached lettuce leaves kept in darkness, revealed that a sharp decline in GAs accompanied by a moderate rise in ABA occurred before the onset of chlorophyll degradation. As senescence advanced, no GAs could be detected and a considerable rise of ABA was observed. A similar sequence of hormonal modifications, but more pronounced, was observed in the course of accelerated senescence induced by either Ethephon or water stress. When kinetin or GA₃ was applied to detached leaves, the loss of chlorophyll and the rise in ABA were reduced. Bound GAs were detected in senescent leaves. They were not found in the kinetin-treated leaves, which contained a relatively high level of free GAs. The results suggest that senescence in detached romaine lettuce leaves is connected with a depletion of free GAs and cytokinins, which is thereafter followed by a great surge in ABA.     

Effects of spermidine on chlorophyll content,photosynthetic activity and chloroplast ultrastructure in the dark and under light

Spermidine prevented the loss of chlorophyll from detached komatsuna (Brassica campestris cv. komatsuna) leaves in the dark but caused a striking reduction in light. It also inhibited photochemical activity in leaves incubated in the dark and light. Striking changes were seen in the chloroplast ultrastructure in light; the envelope was lacking, grana were dilated and thylakoids were distributed throughout the cytoplasm.     

Chlorophyll fluorescence induction,chlorophyll content,and chromaticity characteristics of leaves as indicators of photosynthetic apparatus senescence in arboreous plants

Parameters of chlorophyll fluorescence induction (CFI) are widely used for assessment of the physiological state of higher plant leaves in biochemical, physiological, and ecological studies and in agricultural applications. In this work we have analyzed data on variability of some CFI parameters — Φ _PSII ^max = F _v/F _m (relative value of variable fluorescence), q _NPQ (non-photochemical quenching coefficient), R _Fd (“vitality index”) — in autumnal leaves of ten arboreous plant species of the temperate climatic zone. The correlation between the chlorophyll content in the leaves and fluorescence parameters characterizing photosynthetic activity is shown for two representative species, the small-leaved linden Tilia cordata and the rowan tree Sorbus aucuparia. During the period of mass yellowing of the leaves, the Φ _PSII ^max value can be used as an adequate characteristic of their photochemical activity, while in summer the q _NPQ or R _Fd values are more informative. We have established a correlation between the Φ _PSII ^max value, which characterizes the maximal photochemical activity of the photosystem II, and “chromaticity coordinates” of a leaf characterizing its color features. The chromaticity coordinates determined from the optical reflection spectra of the leaves serve as a quantitative measure of their hues, and this creates certain prerequisites for a visual expert assessment of the physiological state of the leaves.     

Effect of lincomycin on the chlorophyll protein complex I content and Photosystem I activity of greening leaves

1. 1. Greening barley and pea leaves treated with lincomycin have a reduced chlorophyll content. Lincomycin does not alter the proportion of chlorophyll in chlorophyll-protein complex II (CPII) but greatly reduces that in chlorophyll-protein complex I (CPI).

2. 2. Difference spectra show that chloroplasts from lincomycin-treated leaves are deficient in at least two long wavelength forms of chlorophyll a. These have maxima at 77 K of 683 and 690 nm.

3. 3. The chemically determined P-700/chlorophyll ratio of chloroplasts is unaffected by lincomycin but the photochemical P-700/chlorophyll ratio is less than half of that of the control. It is less affected than the chlorophyll-protein complex I content.

4. 4. Photosystem I activity expressed on a chlorophyll basis is unaffected by lincomycin but the light intensity for half saturation is increased 8-fold.

5. 5. Chlorophyll-protein complex I apoprotein content is reduced by lincomycin. No evidence was found for an accumulation of its precursor(s). The relative abundance of major peptides of 18 000, 15 000 and 12 000 daltons in lincomycin-treated chloroplasts is attributed to a general inhibition of greening and associated membrane formation.

Light-stimulation of alanine aminotransferase activity in dark-grown leaves of Lolium temulentum L. as related to chlorophyll formation

Summary The activity of alanine aminotransferase (=glutamate-pyruvate transaminase, GPT) in dark-grown first leaves of Lolium temulentum L. was increased, after an initial lag-phase of 4–6 hr, by more than 130% during the first 24 hr of light-exposure. In comparison, aspartate aminotransferase (=glutamateoxalacetate transaminase, GOT) activity rose by only 18%. Red light treatments of up to 60 min duration produced subsequent increases in GPT activity but the effects were too small to indicate a phytochrome-mediated response. The amounts of enzyme formed were equivalent to those obtained with similar incident intensities of white light. Retuern to darkness after light exposure resulted in an arrestation of the light-stimulated GPT increase. Pre-treatment with cycloheximide caused either stimulatory or inhibitory effects depending upon the concentration applied but, in general, chlorophyll formation and GPT activity responded in a similar manner, whilst GOT showed virtually no response. Chloramphenicol at 6x10^-3 M depressed chlorophyll and Fraction 1 protein synthesis but stimulated GPT activity.The data are discussed in relation to the possible roles of GPT in the leaf. It is suggested that the enzyme, as determined, may be a complex of forms and that at least part of the activity may be involved in the early stages of chlorophyll biosynthesis.     

外源Spd对高温胁迫下黄瓜幼苗叶片膜脂过氧化及质子泵活性的影响

以较为耐热的黄瓜品种‘津春4号’为试材,在人工气候箱中,采用石英砂培养加营养液浇灌的栽培方式,研究了叶面喷施外源亚精胺(Spd)对高温胁迫下黄瓜幼苗叶片膜脂过氧化程度、质子泵活性及其基因表达的影响.结果表明:高温胁迫下,外源Spd促进黄瓜幼苗株高、茎粗、干、鲜质量和叶面积显著增加,有效抑制叶片相对电导率、丙二醛(MDA)含量和脂氧合酶(LOX)活性的升高,有助于提高叶片细胞质膜和液泡膜H+ -ATPase活性,但在基因表达水平上无显著差异.外源Spd可显著降低黄瓜幼苗叶片膜脂过氧化程度,提高质子泵活性,从而稳定膜的结构和功能,缓解高温胁迫对黄瓜幼苗造成的伤害,提高幼苗对高温胁迫的耐性.     

Polyamines and root formation in mung bean hypocotyl cuttings : I. Effects of exogenous compounds and changes in endogenous polyamine content

The effect of several polyamines (putrescine, spermidine, and spermine), their precursors (l-arginine and l-ornithine), and some analogs and metabolic inhibitors (l-canavanine, l-canaline, and methylglyoxal-bis [guanylhydrazone]) on root formation have been studied in mung bean (Vigna radiata [L.] Wilczek) hypocotyl cuttings.     

遮阴对闽楠叶绿素含量和光合特性的影响

为探讨闽楠对不同光环境的光合适应机制,以2年生闽楠幼苗为材料,设置3个光照处理(全光照、遮光率50%和遮光率78%),适应6个月后,测定其叶绿素含量、气体交换和叶绿素荧光同步数据,研究不同光环境处理对闽楠叶片叶绿素含量、叶绿素荧光参数和光合特性的影响.结果表明: 3种光照处理下,闽楠叶片叶绿素a、叶绿素b、叶绿素(a+b)和类胡萝卜素含量大小次序为78%遮光率>50%遮光率>全光照,但不同光照处理对闽楠叶绿素a/b值没有显著影响.遮阴条件下,闽楠叶片光补偿点(LCP)降低,光饱和点(LSP)和表观量子效率(AQY)升高,说明遮阴条件下闽楠叶片对弱光和强光的利用能力均有所提高;最大净光合速率(P_{n max})、光下暗呼吸速率(R_d)和最大电子传递速率(J_max)均增大.在不同处理间,闽楠叶片净光合速率(P_n)、CO₂气孔导度(g_sc)、胞间CO₂浓度(C_i)和叶肉导度(g_m)均存在显著差异.P_n和g_m的大小顺序为: 78%遮光率>50%遮光率>全光照.78%遮光率处理下g_sc显著大于全光照.50%遮光率条件和78%遮光率条件下C_i均显著小于全光照.78%遮光率条件下PSⅡ实际光量子产量(F_v′/F_m′)、PSⅡ光化学效率(Φ_PSⅡ)和电子传递速率(J)均显著大于50%遮光率条件和全光照.由此可知,在遮阴条件下闽楠可以通过增加叶绿素含量、AQY、J、g_sc和g_m来增大光合能力.     

Regulation of tRNA methyltransferase activities by spermidine and putrescine. Inhibition of polyamine synthesis and tRNA methylation by alpha-methylornithine or 1,3-diaminopropan-2-ol in Dictyostelium.

Inhibitors of polyamine synthesis (alpha-methylornithine and 1,3-diaminopropan-2-ol) were used to study the relationship between polyamine synthesis and specific methylations of tRNA in Dictyostelium discoideum during vegetative growth. Polyamine concentrations were found to be 10 mM for putrescine, 1.6 mM for spermidine and 7 mM for 1,3-diaminopropane throughout the growth stage. On treatment of growing amoebae with alpha-methylornithine or with 1,3-diaminopropan-2-ol (each at 5 mM), the syntheses of putrescine, spermidine and 1,3-diaminopropane were arrested within 4h. After polyamine synthesis had ceased, the incorporation of methyl groups into tRNA was considerably decreased under conditions that had no effect on the incorporation of uridine into tRNA, or on net syntheses of protein and of DNA. The following nucleosides in tRNA were concerned: 1 methyladenosine, 5-methylcytidine, 7-methylguanosine, 2-methylguanosine, N2N2-dimethylguanosine and 5-methyluridine (ribosylthymine). The corresponding tRNA methyltransferases, determined in Mg2+-free enzyme extracts, proved to be inactive unless polyamines were added. Putrescine and/or spermidine at concentrations of 10 mM or 1-2 mM respectively stimulate the transmethylation reaction in vitro to a maximal rate and to an optimal extent at exactly the same concentrations as found in vegetative cells. In contrast, 1,3-diaminopropane, which is formed from spermidine, does not affect the methylation of tRNA in vitro at physiological concentrations. Putrescine and/or spermidine stabilize the tRNA methyltransferases in crude extracts in the presence but not in the absence of the substrate tRNA. The results support the view that S-adenosylmethionine-dependent transmethylation reactions can be regulated by alterations of polyamine concentrations in vivo.     

Intensity of photosynthesis and chlorophyll content as related to leaf age inNicotiana Sanderae hort

U r?zně starých list? v listové r??ici 90 a? 110 denních rostlin Nicotiana sanderae hort. byly sledovány rozdály v intensitě ?isté fotosynthesy a v obsahu chlorofylu (a + b). Ke stanovení intensity fotosynthesy bylo pou?ito dvou odli?ných metod, a to váhového stanovení p?ír?stku su?iny podle Barto?e, KubÍna a ?et-lÍka (1960) a gazometrického stanovení infra?erveným analyzátorem CO₂. Nejvy??í intensitu fotosynthesy i nejvy??í obsah chlorofylu (vzhledem k plo?e listové) mají mladé, ale ji? dob?e rozvinuté listy, tj. t?etí a? ?tvrté od vrcholu (prvním listem se rozumí list o plo?e asi 20 cm²). Tyto listy nazýváme ?fotosyntheticky dospělými“. Listy nejmlad?í a zejména pak listy star?í mají intensitu fotosynthesy i obsah chlorofylu ni??í; u nejstar?ích list? je intensita fotosynthesy prakticky nulová. Intensita fotosynthesy i obsah chlorofylu se během vývoje mění: jejich momentální rozdíly u list? v genetické spirále jsou z?ejmě shodné s jejich změnami v ontogenesi listu. Pokles intensity fotosynthesy p?i stárnutí list? je rychlej?í ne? pokles obsahu chlorofylu. P?i ur?itém obsahu chlorofylu (tj. asi 2,25 a? 2,45 mg/dm²) klesá intensita ?isté fotosynthesy k nule. Intensita fotosynthesy je v lineárním vztahu k mno?ství chlorofylu (p?i p?epo?tu na plo?nou jednotku), a to nezávisle na poloze listu v genetické spirále. Obě pou?ité metody ke stanovení intensity fotosynthesy poskytly obdobné výsledky.     

Effects of cadmium on growth, photosynthetic rate and chlorophyll content in leaves of soybean seedlings

Cadmium is a widely spread pollutant and the objective of this study was to study the effects of different concentrations of Cd (0, 50, and 100 μM) on the soybean seedlings treated for 10 d. The growth was inhibited and chlorophyll content, net photosynthetic rate, stomatal conductance, intercellular CO₂ concentration, probability that a trapped exciton moves an electron into the electron transport chain beyond QA — (ET_O/TR_O), performance indexes (PI_ABS), and amount of active reaction centres per excited cross section (RC/CS_O) were decreased with the increasing Cd concentration. The leaf reflectance increased in visual range (500–670 nm) and decreased in near-infrared range (750–1 000 nm). The accumulation of Cd in the roots was much higher than that in the stems and leaves.     

Effects of conditional overexpression of spermidine/spermine N1-acetyltransferase on polyamine pool dynamics, cell growth, and sensitivity to polyamine analogs

Acetylation of polyamines by spermidine/spermine N(1)-acetyltransferase (SSAT) has been implicated in their degradation and/or export out of the cell. The relationship of SSAT to polyamine pool dynamics and cell growth is not yet clearly understood. MCF-7 human breast carcinoma cells were transfected with tetracycline-regulated (Tet-off) SSAT human cDNA or murine gene. Doxycycline removal for >2 days caused a approximately 20-fold increase in SSAT RNA and a approximately 10-fold increase in enzyme activity. After 4 days, intracellular putrescine and spermidine pools were markedly lowered, and cell growth was inhibited. Growth inhibition could not be prevented with exogenous polyamines due to a previously unrecognized ability of SSAT to rapidly acetylate influxing polyamines and thereby prevent restoration of the endogenous pools. Instead, cells accumulated high levels of N(1)-acetylspermidine, N(1)-acetylspermine, and N(1), N(12)-diacetylspermine, a metabolite not previously reported in mammalian cells. Doxycycline deprivation before treatment with N(1), N(11)-diethylnorspermine markedly increased analog induction of SSAT mRNA and activity and enhanced growth sensitivity to the analog by approximately 100-fold. Overall, the findings demonstrate that conditional overexpression of SSAT lowers polyamine pools, inhibits cell growth, and markedly enhances growth sensitivity to certain analogs. The enzyme also plays a remarkably efficient role in maintaining polyamine pool homeostasis during challenges with exogenous polyamines.     

Effect of light and benzyladenine on dark-treated growing rice leaves (Oryza sativa). I. Changes in chlorophyll content and catalase activity

Light- and benzyladenine-induced reversal of the changes in chlorophyll content and catalase activity were studied in the attached first leaf of Oryza sativa L. cv. Bala, kept in darkness for different periods before maturation. Dark treatment caused a decrease in chlorophyll content and catalase activity at all times. Light treatment of dark-incubated seedlings at different periods before maturation reversed the dark-induced effect on chlorophyll content, catalase activity and dry weight and also caused a further rise in chlorophyll content compared to initial values. In darkness, the application of benzyladenine replaced the light effect in maintaining catalase activity. Chlorophyll content was also maintained by initially applied benzyladenine. Benzyladenine did not promote the photoinduced maintenance and increase in chlorophyll content and catalase activity at any time. Treatment with hydrogen peroxide, glycolate and amizol resulted in an accelerated chlorophyll breakdown and had varied effects on catalase levels. Chlorophyll decrease due to peroxide accumulation was to some extent reversible by benzyladenine, but the hormone had no effect on the peroxide-induced decrease in catalase activity. Development of catalase is light dependent. Benzyladenine stabilises the enzyme but has no effect on its synthesis.     

Uptake of diuron and concomitant loss of photosynthetic activity in leaves as visualized by imaging the red chlorophyll fluorescence

The principles of the chlorophyll (Chl) fluorescence induction kinetics (known as Kautsky effect) and their change by the photosystem II herbicide diuron are presented together with the Chl fluorescence emission spectra of a normal and diuron-inhibited leaf. By imaging the Chl fluorescence emission of green leaves the successive uptake of diuron and the concomitant loss of photosynthetic quantum conversion from the leaf base to the leaf tip are documented.     

Polyamines and ”scavenging system”: influence of exogenous spermidine on catalase and guaiacol peroxidase activities,and free polyamine level in barley leaves under water deficit

Barley (Hordeum vulgare) seedlings were treated with spermidine prior to water deficit treatment to determine whether this polyamine is able to influence the activity of catalase (EC 1.11.1.6) and guaiacol peroxidase (EC 1.11.1.7). The content of endogenous polyamines was determined as well. This stress caused a visible increase of catalase activity and a much higher increase of guaiacol peroxidase activity. Treatment with exogenous spermidine induced a significant decrease in the activity of the both enzymes, whereas the polyamine level was enhanced, suggesting that polyamines are able to influence the activity of hydrogen peroxide scavenging enzymes and to moderate in that way this signal molecule level.     

Effects of cysteine on growth, protease production, and catalase activity of Pseudomonas fluorescens.

Cysteine inhibits growth of and protease production by Pseudomonas fluorescens NC3. Catalase activity in P. fluorescens NC3 was increased by cysteine. The addition of exogenous hydrogen peroxide did not increase catalase activity, thus suggesting a role for the endogenous generation of hydrogen peroxide via the autoxidation of cysteine.