Characterization and kinetics of soybean maturation and monocarpic senescence

Daring monocarpic senescence in potted soybeans ( Glycine maxi (L.) Merrill cv. Anoka) grown in controlled-environment chambers, foliar chlorophyll, soluble protein nitrogen, total nitrogen, and starch decline (roughly in that order). All of these precede visible yellowing and, of course, abscission. The pattern of yellowing within a leaf is not uniform and is closely paralleled by starch loss. Unexpectedly, acid-soluble nitrogen rises slightly before the total foliar nitrogen declines. Foliar fresh weight and total dry matter/cm2 of leaf surface decline little if at all before shedding. Preceding and even during the foliar yellowing, the seeds rapidly accumulate dry matter and nitrogen. Yellowing appears first in the radicle tip, then in the rest of the axis and the leaves and finally in the carpels. Ability to germinate is acquired at about the time the radicle + hypocotyl turns yellow. The relationship between these changes and their role in senescence is discussed.     

Effects of moisture stress and senescence on the synthesis of abscisic acid in the primary leaves of bean

The capacity for ABA synthesis during moisture stress of primary leaves of bean ( Phaseolus vulgaris cv. Kinghorn) was defined in terms of leaf age and associated changes in several physiological parameters. The leaves were fully expanded within 9 days after emergence. Fresh and dry weights per unit of leaf area fell during all 5 weeks of the study, from leaf expansion through advanced senescence. The most significant losses in weight occurred during the third and fourth weeks and coincided with a sharp drop in protein content that began immediately after full-leaf. Chlorophyll concentrations declined rapidly during leaf expansion and then more slowly through the end of the fifth week when the leaves were ready to abscise. The ratio of chlorophyll a to b rose steeply over the first 4 weeks of the study.
Although a rapid loss of protein provided the most definitive indication of the early stages of leaf senescence, a marked decline in the ability to synthesize ABA was more closely associated with the termination of rapid leaf growth. This relationship between leaf expansion and the capacity for ABA synthesis during moisture stress remained unchanged when ABA content was expressed on a per unit chlorophyll, protein or dry weight basis.
A water deficit between 5 and 10% of fresh weight, representing a drop in water potential of less than 150 kPa, was sufficient to initiate accumulation of ABA in young leaves. Slightly more intensive levels of stress were required to stimulate ABA synthesis in senescent leaves, but total accumulation was less than one-tenth of the amount recorded in the younger tissue.     

Effects of cotton rootstock on endogenous cytokinins and abscisic acid in xylem sap and leaves in relation to leaf senescence

Leaf senescence varies greatly among cotton cultivars, possiblydue to their root characteristics, particularly the root-sourcedcytokinins and abscisic acid (ABA). Early-senescence (K1) andlate-senescence (K2) lines, were reciprocally or self-graftedto examine the effects of rootstock on leaf senescence and endogenoushormones in both leaves and xylem sap. The results indicatethat the graft of K1 scion onto K2 rootstock (K1/K2) alleviatedleaf senescence with enhanced photosynthetic (Pn) rate, increasedlevels of chlorophyll (Chl) and total soluble protein (TSP),concurrently with reduced malondialdehyde (MDA) contents inthe fourth leaf on the main-stem. The graft of K2 scion ontoK1 rootstock enhanced leaf senescence with reduced Pn, Chl,and TSP, and increased MDA, compared with their respective self-graftedcontrol plants (K1/K1 and K2/K2). Reciprocally grafted plantsdiffered significantly from their self-grafted control plantsin levels of zeatin and its riboside (Z+ZR), isopentenyl andits adenine (iP+iPA), and ABA, but not in those of dihydrozeatinand its riboside (DHZ+DHZR) in leaves in late season, whichwas consistent with variations in leaf senescence between reciprocallyand self-grafted plants. The results suggest that leaf senescenceis closely associated with reduced accumulation of Z+ZR, andiP+iPA rather than DHZ+DHZR, or enhanced ABA in leaves of cotton.Genotypic variation in leaf senescence may result from the differencein root characteristics, particularly in Z+ZR, iP+iPA, and ABAwhich are regulated by the root system directly or indirectly. Key words: Abscisic acid, cotton, cytokinins, grafting, leaf senescence Received 23 October 2007; Revised 17 January 2008 Accepted 23 January 2008     

Regulation of monocarpic senescence of Brassica campestris by the developing pods

Senescence of Brassica campestris L. cv. B-9 was studied with regard to seed maturation and source-sink relationships. In normal control plants leaf senescence (as determined by the change in chlorophyll level) started and proceeded in a progressive manner from base to apex during the period of early pod setting. Complete yellowing of the leaves occurred well before the seed maturation and pod wall senescence. The pod wall always senesced before the attainment of final seed weight. In two different sets of acrocarpous plants containing 65 pods and 10 pods, respectively, leaf senescence was delayed during the pod filling period. It started non-sequentially after complete yellowing and senescence of the pod wall. The degree of leaf senescence at the post-pod filling stage was almost proportional to the number of pods present. When peduncles of the acrocarpous 10-podded plants were removed after the pod filling stage of the plant, leaf senescence was delayed compared to plants whose pedicels were removed, although the senescence pattern of the upper three leaves was nonsequential in both cases. Defruiting at an early stage of development delayed leaf senescence, although the pattern of such senescence remained unaltered (i.e. nonsequential). Defoliation hastened the seed-filling process and pod wall senescence. Plants containing fewer pods had higher average seed weight, although yield per plant was reduced.
These results suggest that the pod wall serves as a temporary as well as intermediary storage organ and that foliar senescence is not directly related to seed maturation. The possible cause of uncoupling between foliar senescence and seed development is discussed.     

Incorporation of C-Leucine into Apple Leaf Protein and Its Inhibition by Protein Synthesis Inhibitors during Growth and Senescence

Of the total ¹⁴C-leucine taken up by intact apple (Pyrus malus L., Golden Delicious) leaf discs, 44 to 62% is incorporated into protein from June to early October. Of this amount, an average of 35% is released by mild, room temperature acid hydrolysis. Prior to mid-August when leaf protein begins to decline, 15 to 20% of the ¹⁴C-leucine incorporated into protein occurs in water-(buffer) soluble protein, of which only 3% is released by mild acid hydrolysis. After mid-August, 40% of the label in protein occurs in soluble protein. The specific radio-activity of the soluble protein increases by 4- to 5-fold after mid-August, while that of total protein increases by less than 2-fold. In presenescent leaves (before the decline of protein in August) 20 micrograms per milliliter cycloheximide inhibits the incorporation of ¹⁴C-leucine into protein by 71%, and 20 micrograms per milliliter chloramphenicol inhibits it by 30%. In senescing leaves, cycloheximide inhibits ¹⁴C-leucine by 85% or more, while chloramphenicol inhibits it by less than 15%. Coincident to the initial decline of leaf protein, chloramphenicol greatly loses its ability to inhibit the incorporation of ¹⁴C-leucine into apple leaf protein. At all leaf ages, chloramphenicol increases the loss of chlorophyll from apple leaf discs. The effect of cycloheximide on leaf disc senescence changes with leaf age: in early season samples, it increases the loss of chlorophyll; in mid-season samples, it has no effect; and in late season samples, it retards the loss of chlorophyll.     

Observations on the Senescence of a Mutant Non-yellowing Genotype of Phaseolus vulgaris L.

A non-yellowing mutant of Phaseolus vulgaris L. was used toinvestigate factors involved in chlorophyll breakdown duringfoliar senescence. The mutant showed physiological changes similarto those of the normal yellowing type during senescence exceptthat leaf chlorophyll did not decline. Transmission electronmicroscope studies did not reveal appreciable differences inchloroplast ultrastructure between the two genotypes, suggestingthat chloroplast membrane integrity was not the factor preventingchlorophyll degradation in the mutant. However, the lack ofplastoglobuli in senescent mutant chloroplasts suggested thatthe lipid environment may be different from that of senescentnormal chloroplasts. Banding patterns of total soluble protein,resolved by sodium dodecyl sulphate-poly aery lamide gel electrophoresisshowed few, if any, differences between mature non-senescentnormal and mutant leaves; however, bands at 14 kD and 58 kDdiminished in senescent normal leaves, but remained in senescentmutant non-yellowing leaves. Key words: Non-yellowing mutant, Phaseolus vulgaris, senescence, chlorophyll degradation     

Physiological and Molecular Changes of Detached Wheat Leaves in Responding to Various Treatments

Leaf senescence is induced or accelerated when leaves are detached. However, the senescence process and expression pattern of senescence-associated genes (SAGs) when leaves are detached are not clearly understood. To detect senescence-associated physiological changes and SAG expression, wheat (Triticum aestivum L.) leaves were detached and treated with light, darkness, low temperature (4 C), jasmonic acid (JA), abscisic acid (ABA), and salicylic acid (SA). The leaf phenotypes, chlorophyll content, delayed fluorescence (DF), and expression levels of two SAGs, namely, TaSAG3 and TaSAG5, were analyzed. Under these different treatments, the detached leaves turned yellow with different patterns and varying chlorophyll content. DF significantly decreased after the dark, ABA, JA and SA treatments. TaSAG3 and TaSAG5, which are expressed in natural senescent leaves, showed different expression patterns under various treatments. However, both TaSAG3 and TaSAG5 were upregulated after leaf detachment. Our results revealed senescence-associated physiological changes and molecular differences in leaves, which induced leaf senescence during different stress treatments.     

Identification of a novel E3 ubiquitin ligase that is required for suppression of premature senescence in Arabidopsis

During leaf senescence, resources are recycled by redistribution to younger leaves and reproductive organs. Candidate pathways for the regulation of onset and progression of leaf senescence include ubiquitin‐dependent turnover of key proteins. Here, we identified a novel plant U‐box E3 ubiquitin ligase that prevents premature senescence in Arabidopsis plants, and named it SENESCENCE‐ASSOCIATED E3 UBIQUITIN LIGASE 1 (SAUL1). Using in vitro ubiquitination assays, we show that SAUL1 has E3 ubiquitin ligase activity. We isolated two alleles of saul1 mutants that show premature senescence under low light conditions. The visible yellowing of leaves is accompanied by reduced chlorophyll content, decreased photochemical efficiency of photosystem II and increased expression of senescence genes. In addition, saul1 mutants exhibit enhanced abscisic acid (ABA) biosynthesis. We show that application of ABA to Arabidopsis is sufficient to trigger leaf senescence, and that this response is abolished in the ABA‐insensitive mutants abi1‐1 and abi2‐1, but enhanced in the ABA‐hypersensitive mutant era1‐3. We found that increased ABA levels coincide with enhanced activity of Arabidopsis aldehyde oxidase 3 (AAO3) and accumulation of AAO3 protein in saul1 mutants. Using label transfer experiments, we showed that interactions between SAUL1 and AAO3 occur. This suggests that SAUL1 participates in targeting AAO3 for ubiquitin‐dependent degradation via the 26S proteasome to prevent premature senescence.     

Plant Growth Regulators and Induction of Leaf Senescence in Nitrogen-Deprived Wheat Plants

The sequence of events and the signals that regulate the remobilization of nitrogen (N) reserves during senescence induced by N starvation were studied in leaf 3, the last fully expanded leaf, in 17-day-old wheat (Triticum aestivum L.) plants. The first event observed was a rapid decrease in the isopentenyl adenosine (iPA) concentration during the first 24 h of N starvation. No differences in t-zeatin riboside and dihydrozeatin riboside concentrations were observed until the end of the assay. During the following 6 days, a decrease in soluble amino acids, chlorophyll, and protein, as well as an increase in soluble sugar concentration and endoproteolytic activity, could be observed. At day 3 of the experiment, the abscisic acid (ABA) concentration in the leaves of N-deprived plants started to increase. After 6 days of N deprivation there was a rise in oxidative stress, as indicated by the increase in malondialdehyde concentration, as well as a decrease in the activities of antioxidant enzymes catalase and ascorbate peroxidase. To analyze interactions with leaf development, the first, second, third, and fourth leaves were studied. iPA concentration decreased in all the leaf stages, including leaf 4, which was not fully expanded. A linear correlation between iPA and protein concentration was determined. These results suggest that the sharp fall in iPA could be the earliest event that induces protein degradation during the development of senescence induced by N deficiency, and that only later is ABA accumulated and oxidative stress developed.     

棉花叶片衰老过程中激素和膜脂过氧化的关系

以陆地棉品种辽棉9号的去根幼苗为材料,对其进行暗诱导衰老培养.在培养液中分别加入6-BA、ABA、GSH、H2O2、CaCl2、A23187 和A23187 CaCl2,测定在不同培养条件下棉花去根幼苗叶片内源激素、SOD酶活性和MDA含量的变化.结果表明:棉花叶片衰老表现为细胞分裂素含量的下降和ABA含量的上升.6-BA、GSH和钙离子均延缓叶片的衰老,ABA和H2O2促进叶片的衰老.     

Interaction of Abscisic Acid and Light on Leaf Senescence in Rice

The senescence of excised leaves of Oryza saliva L. cv. BAM11 was studied by monitoring the breakdown of chlorophyll andprotein. ABA at 10^–6 M retarded senescence until the 3rdday and accelerated it in a normal way until the 9th day inlight. The effect of ABA was light-dependent, which is beingreported for the first time in rice. ABA in the presence oflight (26.3 Klux) delayed protein breakdown but could not preventthe yellowing effect. (Received June 18, 1982; Accepted January 28, 1983)     

Leaf senescence in a non-yellowing cultivar of chrysanthemum (Dendranthema grandiflora)

Chlorophyll (Chl) and total soluble protein decreased and proteolytic activity increased over a 12-day period during dark-induced senescence in detached leaves of Tara, a yellowing cultivar (Y) of Dendranthema grandiflora . In Boaldi, a non-yellowing cultivar (NY), Chl and soluble protein remained near initial levels and little change in proteolytic activity was observed. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of soluble proteins showed no major differences in banding patterns between the two cultivars at day 0; however, all of the resolved proteins were diminished in Tara by day 12. On the other hand, in NY Boaldi, the intensity of the protein bands did not change over the 12-day period. Attached and detached leaves exhibited similar senescence patterns for each cultivar. Ethylene (100 μl l⁻¹) accelerated the rate of Chl loss in detached leaves of Tara, but had no effect on Boaldi. These observations suggest that Boaldi is a stay-green genotype, possibly a functional type. The results are discussed in relation to the role of ethylene in chrysanthemum leaf senescence.     

Hormonal changes associated with leaf senescence in cotton (Gossypium barbadense)

To study the hormonal regulation of foliar senescence in cotton ( Gossypium barbadense L. cv. Giza 68, long staple), the sequential changes in gibberellins (GAs), indoleacetic acid (IAA) and abscisic acid (ABA) were examined in the cotyledons from the completion of expansion through senescence (days 12-24 after sowing). The onset of senescence could be detected on day 20, the stage of maximum accumulation of leaf metabolites. At this stage, free GAs quickly lost more than 40% of their initial activity. Further decrease of free GAs was then characteristically observed in the senescent leaves. A remarkable increase in free IAA and free ABA between days 18 and 20 immediately followed by nutrient depletion, suggests the contribution by both hormones to the senescence system. The definite drop in free IAA below its initial level occurred on day 24, when most of the leaf protein and chlorophyll were already broken down.     

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.     

Expression of photosynthesis- and senescence-related genes during leaf development and senescence in silver birch (Betula pendula) seedlings

Leaf development and senescence were studied in greenhouse-grown silver birch ( Betula pendula Roth) seedlings over a period of 7 weeks. Prior to the experiment, leaves from 100 seedlings were marked for five sampling dates. Timing of the developmental phases was studied with biochemical analyses of total soluble protein, Rubisco protein, chlorophyll concentration and at the level of gene expression related to photosynthesis, energy metabolism, ethylene synthesis and protein degradation. During the sampling period, an initial increase in photosynthetic capacity could be seen, when expression of the Rubisco small subunit gene ( RbcS ) and Rubisco protein (EC 4.1.1.39) were examined. Down-regulation of photosynthesis, visible as a decrease in Rubisco protein and RbcS mRNA, started soon after full expansion of the leaves and processes related to senescence followed. mRNA accumulation for the ethylene-forming enzyme 1-aminocyclopropane-1-carboxylic acid oxidase increased first during the onset of senescence. Protein degradation was observed as a loss of soluble proteins, with a simultaneous increase in the leucine aminopeptidase (EC 3.4.11.1) mRNA levels. The mRNA levels of ribonuclease-like pathogenesis-related protein 10 also increased clearly during senescence, but the mitochondrial phosphate translocator mRNA showed only a slight increase. Chlorophyll concentration of the leaves decreased after the mRNA levels of these senescence-related genes had become more abundant.     

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 Effect of Light and Hormones on Leaf Senescence

With wheat leaves as material, the changes of superoxide dismutase (SOD), lipid peroxi-dation and membrane permeability during leaf senescence in light or dark, and treated withphytohormones (KT or ABA) have been studied. The changes of chlorophyll content, lipidperoxidation and fine structure of spinach chloroplasts senescing in light or dark have alsobeen studied. When leaves senesce in light, the activity of SOD increased at first then decreased. The increase of SOD activity was able to result from the synthesis of new protein. Lightwas found to delay the leaf senescence obviously but also accelerate leaf senescence by causinglipid peroxidation when prolonged the illumination time. The delay or acceleration of leafsenescence by exogenous hormones were observed, it may be due to the control of lipid peroxi-dation by adjusting the activity of SOD. O2-participated the chlorophyll decomposition andlipid peroxidation during chloroplasts senesce in light. A favourable role of light in mainta-lng the fine structure of isolated chloroplasts was clear.     

What stay-green mutants tell us about nitrogen remobilization in leaf senescence

Leaf senescence has an important role in the plant's nitrogen economy. Chlorophyll catabolism is a visible symptom of protein mobilization. Genetic and environmental factors that interfere with yellowing tend to modify protein degradation as well. The chlorophyll-protein relationship is much closer for membrane proteins than it is for soluble or total leaf proteins. In stay-greens, genotypes with a specific defect in the chlorophyll catabolism pathway, soluble protein degradation during senescence may be close to normal, but light-harvesting and reaction centre thylakoid membrane proteins are much more stable. Genes for the chlorophyll catabolism pathway and its control are important in the regulation of protein mobilization. Genes for three steps in the pathway are reported to have been isolated. The gene responsible for the stay-green phenotype in grasses and legumes has not yet been cloned but a fair amount is known about it. Pigment metabolism in senescing leaves of the Festuca-Lolium stay-green mutant is clearly disturbed and is consistent with a blockage at the ring-opening (PaO) step in chlorophyll breakdown. PaO is de novo synthesized in senescence and thought to be the key enzyme in the chlorophyll a catabolic pathway. The stay-green mutation is likely to be located in the PaO gene, or a specific regulator of it. These genes may well be in the various senescence-enhanced cDNA collections that have been generated, but functional handles on them are currently lacking. When the stay-green locus from Festuca pratensis was introgressed into Lolium temulentum, a gene encoding F. pratensis UDPG-pyrophosphorylase was shown to have been transferred on the same chromosome segment. A strategy is described for cloning the stay-green gene, based on subtractive PCR-based analyses of intergeneric introgressions and map-based cloning.     

Effect of head removal on leaf senescence of sunflower

Greenhouse and field studies examined the effect of flower or seedhead removal on leaf senescence and associated changes in sunflower (Helianthus annuus L.) plants. At intervals during seed development, selected leaves (leaves 6 through 8 from the top in the greenhouse and leaf 7 from the top in the field) were harvested and analyzed for chlorophyll, specific leaf weight, N, P, soluble protein, and electrophoretic gel profiles of soluble polypeptides. In both the greenhouse and the field, the leaves of headless plants retained or accumulated more N, P, soluble protein, and dry weight than leaves of plants with heads. Obviously, head removal affected the partitioning of these metabolites during seed development. None of the treatments resulted in the formation of new polypeptides (electrophoretic gel profiles). Comparisons of the rates and extent of loss of chlorophyll, soluble protein, and polypeptide bands (especially ribulose 1,5-bisphosphate carboxylase) from the leaves of headed and deheaded plants showed that head removal delayed the rate of development of leaf senescence for the greenhouse-grown but had much less effect on field-grown plants. These findings illustrate the variability in different parameters commonly associated with the leaf senescence processes of headed and deheaded sunflower plants grown under different environments.