Probing Monocarpic Senescence and Pod Development Through Manipulation of Cytokinin and Mineral Supplies in Soybean Explants

Defined solutions containing cytokinin and/or mineral nutrientswere supplied in lieu of the roots through the cut stem baseof soybean explants (a leaf with associated pod and subtendingstem segment) in order to analyze the roles of cytokinin andmineral nutrients from the roots in pod development and foliarmaintenance. In explants cut at early-mid podfill, supplyingonly H₂O accelerated leaf senescence and pod maturation anddecreased seed d. wt relative to comparable parts of intactplants. Zeatin (Z) and/or minerals not only delayed leaf yellowingand the decline in foliar chlorophyll levels and photosyntheticrates but also inhibited leaflet and petiole abscission relativeto H₂O controls. Even large declines in foliar assimilatoryprocesses did not necessarily lead to abscission. Z and/or mineralsalso increased stomatal conductivity throughout podfill. Z showedsome positive synergistic effects with minerals on leaf maintenance.Pod wall, cotyledon and radicle yellowing were delayed by Zand/or minerals but not as much as leaf senescence. Mineralsonly or Z +minerals prolonged seed d. wt accumulation and increasedfinal dry seed wt to a level similar to that for intact plants.Seed growth showed a complex interrelation with pod wall andleaf f. wt and d. wt changes. A decline in cytokinin and mineralflux from the roots appears to be important for pod-inducedleaf senescence; however, pod development, foliar senescenceand their component processes may be affected differently. Thus,even though the explant is a physiological/nutritional moduleof the whole plant, it is influenced by cytokinin and mineralsfrom the roots and therefore only semiautonomous. Glycine max L. Merr. cv. Anoka, soybean, abscission, cytokinin, chlorophyll, mineral nutrients, seed development, semiautonomous physiological modules, senescence, stomatal resistance     

Effects of Nuclear and Cytoplasmic Genes Altering Chlorophyll Loss on Gas Exchange during Monocarpic Senescence in Soybean

Several genes can alter the yellowing processes that normallyoccur during pod development and monocarpic senescence in soybean.CytG and d₁ + d₂ cause the leaves and seeds to stay green atmaturity. G blocks yellowing of the seed coat but not the leavesor embryos. By contrast, another gene, y₃, causes earlier yellowingof the leaves. This paper examines the effects of these geneson photosynthesis and some related parameters of the senescencesyndrome in near-isogenic backgrounds (cv. Clark). Neither cytGnor d₁ d₂ delays the decline in photosynthetic rate during monocarpicsenescence relative to Clark; while Gd₁ d₂ does. Again, y₃ causesan earlier decrease. Similarly, neither cytG nor d₁ d₂ altersthe decline in stomatal conductance and transpiration, whichoccurs at the end of podfill; however, Gd₁ d₂ delays it andy₃ advances it. Neither cytG nor d₁ d₂ prevents the rise inintercellular CO₂ during podfill, while Gd₁ d₂ does, and y₃promotesit. These changes in intercellular CO₂ may reciprocally reflectthe status (maintenance for Gd₁ d₂ and early loss for y₃) ofthe photosynthetic enzymes. Gd₁ d₂, d₁ d₂ and cytG do not appreciablyaffect leaf blade abscission; however, y₃ prevents it. Gd₁ d₂increases the dry weight seed yield, and y₃ decreases it. Gd₁d₂ inhibits several components in the senescence process, whilecytG seems to affect mainly chlorophyll loss and y₃ may actmore broadly than just accelerating senescence. Clearly, thestay-green trait can be caused by more than one genetic locus,and it may or may not maintain photosynthesis. ³Present address: Instituto de Fisologia Vegetal, Facultad deAgronomia, c.c. 31, 1900-La Plata, Argentina (Received May 18, 1990; Accepted September 4, 1990)     

Studies on Genetic Male-Sterile Soybeans : III. The Initiation of Monocarpic Senescence

Soybean (Glycine max [L.] Merr.) germplasm, isogenic except for loci controlling male-sterility (ms₁) and nodulation (rj₁) was utilized to investigate the effects of reproductive tissue development and nitrogen source on the initiation of monocarpic senescence. The experimental genotypes (Ms₁Rj₁, Ms₁rj₁, ms₁Rj₁, and ms₁rj₁, were selected from a cross between N69-2774 and N59-5259, and were inbred to the F₅ generation. Green-house-grown plants were collected during the period of flowering (77 days after transplanting) until maturity (147 days after transplanting). Leaf tissues from the respective genotypes were analyzed at the various harvest dates for RNA, phenolic, and chlorophyll concentrations; acid protease activity; polypeptide banding patterns of chloroplast thylakoids; and chloroplastic ultrastructure.

Regardless of nitrogen source, total chlorophyll concentrations declined between 77 and 119 days after transplanting, resulting in a 40% loss of chlorophyll per square centimeter in all genotypes. Leaf chlorophyll levels continued to decline at a constant rate in male-fertile genotypes, but remained at a constant level (26 micrograms chlorophyll per square centimeter) in male-sterile genotypes, for the remainder of the study. With increased leaf age, a gradual disruption of thylakoid structures was observed, particularly in chloroplasts from the male-fertile genotypes. Chloroplasts from the male-sterile genotypes appeared to lose starch grains but increased their number of chloroplastic lipid bodies with leaf aging. These data suggest that monocarpic senescence in soybeans was initiated at or before flowering. Although reproductive tissue development probably augmented the process, the response attributed to seed formation was not apparent until the mid-pod fill stage (119 days after transplanting). All genotypes had similar changes in other cellular components that are recognized as indicators of plant senescence regardless of whether the plants produced seed.

     

Dysfunctional Telomeres at Senescence Signal Cell Cycle Arrest via Chk2

Loss of telomere integrity can have two outcomes with opposite predicted effects on tumorigenesis. On the one hand, shortened telomeres in normal cells may trigger cell cycle arrest, leading to tumour suppression. On the other hand, in a tumour cell in which neither the p53 nor pRb pathway is intact, shortened telomeres could initiate chromosome instability and promote tumorigenesis A major issue in telomere research is to understand how shortened dysfunctional telomeres can regulate the onset of cellular senescence. Recent studies have revealed that critically shortened or acutely uncapped telomeres share molecular features with damaged DNA. We have recently linked the phosphorylation and activation of one major DNA damage effector checkpoint kinase, Chk2, to telomere erosion in signalling cell cycle arrest in normal fibroblasts. Here, we discuss several hypotheses to explain the molecular events occurring at shortened telomeres that ultimately lead to cell cycle arrest or increased genomic instability.     

Leaf Senescence Induced by Mild Water Deficit Follows the Same Sequence of Macroscopic, Biochemical, and Molecular Events as Monocarpic Senescence in Pea

We have compared the time course of leaf senescence in pea (Pisum sativum L. cv Messire) plants subjected to a mild water deficit to that of monocarpic senescence in leaves of three different ages in well-watered plants and to that of plants in which leaf senescence was delayed by flower excision. The mild water deficit (with photosynthesis rate maintained at appreciable levels) sped up senescence by 15 d (200 degrees Cd), whereas flower excision delayed it by 17 d (270 degrees Cd) compared with leaves of the same age in well-watered plants. The range of life spans in leaves of different ages in control plants was 25 d (340 degrees Cd). In all cases, the first detected event was an increase in the mRNA encoding a cysteine-proteinase homologous to Arabidopsis SAG2. This happened while the photosynthesis rate and the chlorophyll and protein contents were still high. The 2-fold variability in life span of the studied leaves was closely linked to the duration from leaf unfolding to the beginning of accumulation of this mRNA. In contrast, the duration of the subsequent phases was essentially conserved in all studied cases, except in plants with excised flowers, where the degradation processes were slower. These results suggest that senescence in water-deficient plants was triggered by an early signal occurring while leaf photosynthesis was still active, followed by a program similar to that of monocarpic senescence. They also suggest that reproductive development plays a crucial role in the triggering of senescence.     

Cytotoxic Signal Transmission Pathways via TNF Family Receptors

Studies indicating an important role of the TNF-receptor family in control of cell proliferation, differentiation, and death have drastically increased in number in recent years. The main function of many members of this family is cell death triggering, and this is apparently the only function for some of them. Studies on the molecular mechanisms of cell death activated by members of the TNF-receptor family revealed and identified proteins directly or indirectly associated with TNF-receptors. Pathways of cytotoxic signal transduction by some members of the TNF-Rs family based on currently proven protein–protein interactions and the role of distinct proteins in these processes are summarized in this review.     

Vacuole Cysteine Proteases and Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Degradation during Monocarpic Senescence in Cowpea Leaves

Characterisation of proteases degrading ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO, EC: 4.1.1.39) was studied in the cowpea leaf during monocarpic senescence 3 and 9 d after flowering (DAF), representing early and mid pod fill. The stage at 3 DAF coincided with decrease in the metabolic parameters characterising senescence, i.e., contents of total soluble proteins, RuBPCO, and leaf nitrogen. At 9 DAF, there was a decline in total soluble proteins and an appearance of a 48 kDa cysteine protease. Characterisation of the proteases was done using specific inhibitors. Subcellular localisation at 3 DAF was studied by following the degradation of RuBPCO large subunit (LSU) in the vacuole lysates using immunoblot analyses. Cysteine proteases played a predominant role in the degradation of RuBPCO LSU at the crude extract level. At 9 DAF, expression of cysteine protease isoforms was monitored using polyclonal antibodies against papain and two polypeptides of molecular masses 48 and 35 kDa were observed in the vacuole lysates. We confirmed thus the predominance of cysteine proteases in the vacuoles during different stages of pod development in cowpea leaf.     

大豆子叶衰老的细胞学观察

以萌发后不同时期的大豆子叶为材料,通过普通光镜和荧光显微镜观察,分析了不同时期子叶细胞的结构变化及子叶细胞内蛋白质和淀粉含量的组织化学变化.结果表明,随着种子萌发时期的延长,子叶细胞内的蛋白质和淀粉含量逐渐减少,在子叶衰老过程中,细胞内蛋白质首先消耗殆尽,淀粉的消耗速度较蛋白质慢;大豆子叶细胞在萌发后第18天时出现典型的植物编程性死亡的形态学特征,子叶细胞内营养物质的消耗诱发子叶细胞发生细胞凋亡.     

The Importance of Roots in Regulating the Senescence of Soybean Primary Leaves

The role of roots in regulating primary leaf senescence of 14-day-old soybean seedlings was investigated. Compared with intact seedlings, the senescence of primary leaves is accelerated by removal of the root system but delayed if apical bud and the first trifoliate leaf are removed. No difference in senescence was found between intact seedlings and seedlings without roots, apical bud, and first trifoliate leaf. Lateral roots seem to play a predominant role in regulating primary leaf senescence. However, neither root nodules nor primary root play any function in senescence. Results indicate that benzyladenine (BA) at optimal concentration (2 mg/1) completely replaces the roots to prevent the senescence of primary leaves, whereas gibberellic acid (GA) and abscisic acid (ABA) accelerate. The effect of indole-3-acetic acid (IAA) to replace roots in preventing senescence depends on the season the young seedlings are grown. Additional, though indirect, information of acropetal transport of ABA is provided. In conclusion, it seems that cytokinins in lateral roots play a predominant role in leaf senescence and the normal supply of root cytokinins is important in leaf metabolism.     

Leaf Senescence and Abscisic Acid in Leaves of Field-grown Soybean

Leaf senescence in field-grown soybean (Merrill) as defined by the period after full expansion, was studied by measuring abscisic acid (ABA), total soluble protein, and chlorophyll in leaves through the later part of the growing season. ABA concentrations increased significantly at the end of the season when leaves had started to turn yellow, well after total soluble protein and chlorophyll had started to decline. The results indicate that events occurring before leaf yellowing are more significant in evaluating leaf senescence since the yellowing condition and rise in ABA are effects of changes in physiological activity beginning when leaves are still green.     

Ribulose-1,5-bisphosphate Carboxylase/Oxygenase Content and Degradation in Diploid,Tetraploid, and Hexaploid Wheat Species during Monocarpic Senescence

Wheat provides a unique genetic system in which variable sink size is available across the ploidies. We characterized monocarpic senescence in diploid, tetraploid, and hexaploid wheat species in flag leaf from anthesis up to full grain maturity at regular intervals. Triticum tauschii Acc. cv. EC-331751 showed the fastest rate of senescence among the species studied and the rate of loss per day was highest in terms of photosynthesis rate, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) content, and flag leaf N content coupled with a higher rate of gain in grain N content. Cultivars Kundan and HD 4530 maintained high flag leaf N content throughout grain filling as compared to the diploids and showed a slower rate of senescence. RuBPCO content was higher in the diploids as compared to Kundan and HD 4530 at anthesis. However, the rate of decline in RuBPCO content per day was also higher in the diploids. This degradation in RuBPCO was mediated by high endoproteolytic activities in the diploids which in turn supported its higher rate of N mobilization as compared to the tetraploid and hexaploid wheat. Acidic endopeptidases were responsible for the mobilization of flag leaf nitrogen in wheat across ploidy levels (r=–0.582, p<0.01).     

Graft Transmission of Longday-induced Leaf Senescence in Kleinia articulata

Kulkarni, V. J. and Schwabe, W. W. 1985 Graft transmission oflongday-induced leaf senescence in Kleinia articulata.—J.exp. Bot. 36: 1620–1633. Senescence of attached and detached rooted leaves in Kleiniaarticulata can be regulated by daylength, accelerated by longphotoperiods LD (> 16 h) and retarded by short days SD (8h). Using detached, rooted leaves as stocks in leaf to leafgrafts, senescence was readily transmitted from a LD donor leafto a SD receptor leaf even if the receptor was retained in SD(D8).However, no transfer could be detected where it had to passthrough any stem tissue. Senescence was reversible up to a certain stage, beyond whichgrafts senesced, pointing to an accumulation of the graft transmissiblefactor to a threshold level that causes irreversible death. Key words: Kleinia, leaf senescence, senescence factor, daylength, graft transmission     

大豆下胚轴线粒体的衰老与膜脂的过氧化作用

离体的大豆下胚轴线粒体,在人工衰老条件下,产生了结构膨胀和细胞色素氧化酶活性的下降。衰老的线粒体也发生膜脂的过氧化作用——丙二醛、脂质的氢过氧化物和荧光脂褐色素明显增加。而且,线粒体衰老时产生的膜脂过氧化产物雨二醛,可能是膜脂的磷脂酰胆碱和磷脂酰乙醇胺中的亚麻酸发生过氧化反应的结果。     

Bacteroids Are Stable during Dark-Induced Senescence of Soybean Root Nodules

Physiological and biochemical markers of metabolic competence were assayed in bacteroids isolated from root nodules of control, dark-stressed, and recovered plants of Glycine max Merr. cv `Woodworth.' Nitrogenase-dependent acetylene reduction by the whole plant decreased to 8% of control rates after 4 days of dark stress and could not be detected in plants dark stressed for 8 days. However, in bacteroids isolated anaerobically, almost 50% of initial acetylene reduction activity remained after 4 days of dark stress but was totally lost after 8 days of dark stress. Bacteroid acetylene reduction activity recovered faster than whole plant acetylene reduction activity when plants were dark stressed for 8 days and returned to a normal light regimen. Significant changes were not measured in bacteroid respiration, protein content, sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles, or in bacteroid proteolytic activity throughout the experiment. Immunoblots of bacteroid extracts revealed the presence of nitrogenase component II in control, 4-day dark-stressed, and 8-day dark-stressed plants that were allowed to recover under a normal light regimen, but not in 8-day dark-stressed plants. Our data indicate that dark stress does not greatly affect bacteroid metabolism or induce bacteroid senescence.     

木质部液流的碱化：一个综合的干旱胁迫响应信号

土壤干旱会改变植物木质部液流的离子组成,引起木质部碱化,提高液流的pH值。因而认为木质部液流碱化是诱使叶上气孔关闭的最可能的根源土壤干旱信号。本文章介绍了这一领域的研究进展。     

Long-Range Signal Transmission in Autocrine Relays

Intracellular signaling induced by peptide growth factors can stimulate secretion of these molecules into the extracellular medium. In autocrine and paracrine networks, this can establish a positive feedback loop between ligand binding and ligand release. When coupled to intercellular communication by autocrine ligands, this positive feedback can generate constant-speed traveling waves. To demonstrate that, we propose a mechanistic model of autocrine relay systems. The model is relevant to the physiology of epithelial layers and to a number of in vitro experimental formats. Using asymptotic and numerical tools, we find that traveling waves in autocrine relays exist and have a number of unusual properties, such as an optimal ligand binding strength necessary for the maximal speed of propagation. We compare our results to recent observations of autocrine and paracrine systems and discuss the steps toward experimental tests of our predictions.     

衰老及相关疾病与细胞信号转导的关系

概述了衰老及相关病变与细胞信号转导障碍或异常的密切关系。细胞与细胞间或细胞与环境间无时无刻不在进行着信息联系 ,信号转导途径的任何一个环节出现问题 ,都可引起细胞生长失控 ,进而引起病变。因而了解细胞衰老机理和信号转导途径 ,从分子水平上认识、预防和治疗疾病有着深远的意义。     

The Effect of Irradiance on the Recovery of Soybean Nodules from Sodium Chloride-Induced Senescence

Soybean (Glycine max L. Merr) cv. Clarke plants inoculated withBradyrhizobium japonicum strain RCR3407 were grown either ina greenhouse with a low irradiance (200–400)µmolm^–2 s^–1) or in a controlled-environment growth cabinetwith a higher irradiance (600 µimol m^–2 s^–1).At 42 d plants were given a nitrogen-free nutrient solutioncontaining 50 mol m^–3 sodium chloride for 2 weeks andthen allowed to recover from salt-stress for a further 2 weeks. Salt treatment reduced plant growth by at least half in bothgrowth regimes, however, the controlled environment-grown (CEG)plants were five times larger than the greenhouse-grown (GG)plants in terms of dry weight and number/weight of nodules perplant, regardless of treatment. The structure of nodules, from both growth regimes, harvestedat the end of the 2 week salt-stress was similar to unstressedcontrol nodules. However, nodules harvested 1 week later fromboth CEG and GG plants had structural changes including degradationof bacteria in vacuoles around host cell nuclei, particularlyin the outer cell layers of the infected tissue. In addition,meristematic activity was seen in the cortex of some nodulesfrom GG plants. Young cells here contained infection threadsand newly-released bacteria. Nodules harvested 2 weeks after removal of the salt-stress fromCEG plants showed an apparent recovery from the stress. However,there was a very marked increase in the amount of starch inthe cortex which was not seen in equivalent GG nodules. In contrast,nodules from GG plants contained many vacuolate infected cellsand, consequently, a lowered bacteroid population. Further,meristematic activity was seen in a zone concentric to the infectedzone, newly-formed cells contained many large infection threadsand were interspersed with intercellular bacteria. The meristematicactivity increased the relative volume of cortical to infectedcells in these nodules. Growth conditions did not affect control nodule specific nitrogenaseactivity or oxygen diffusion resistance (R) and these parameterswere also not altered in CEG nodules exposed to salt plus the14 d recovery period. However, nitrogenase activity was greatlyreduced, and R increased by more than eight times in equivalentGG nodules exposed to salt plus recovery. It is hypothesized that the gross morphological changes werean attempt to counter salt toxicity and/or oxygen damage underconditions of reduced photosynthate supply to the nodules dueto the poor light levels in the greenhouse. However, soybeannodules supplied with adequate photosynthate were able to withstandand recover from long-term salt-stress with little alterationto their structural integrity. Key words: Soybean, sodium chloride, nitrogen fixation, light intensity, oxygen diffusion resistance     

Photosynthesis and Other Traits in Relation to Chloroplast Number during Soybean Leaf Senescence

Soon after attaining full expansion, soybean (Glycine max [L.] Merr.) leaves enter a senescence phase marked by decline in photosynthetic rate and the progressive loss of chloroplast activity and composition. Our primary goal was to determine if this loss could be accounted for by sequential degradation of whole chloroplasts or by simultaneous degeneration of all chloroplasts. Total photosynthesis (TPs) measured as ¹⁴CO₂ uptake, chloroplast number, ribulose 1,5-bisphosphate carboxylase activity, uncoupled photosynthetic electron transport activity, soluble protein content, and chlorophyll content declined progressively during the 37 days after full leaf expansion. During this period, chloroplast number per unit leaf area was constant for all genotypes studied. We conclude that leaf senescence may be a two-stage process wherein the first stage chloroplast activity and composition declines, but chloroplast numbers do not change. During a brief terminal stage (11 days in our experiment), whole chloroplasts may be lost as well. As a second objective we wished to determine if variation in single-leaf total photosynthetic rate among soybean cultivars is related to corresponding variation in chloroplast number and/or chloroplast activity/composition. By comparing the means for three cultivars known to have rapid leaf TPs and for the three known to have slow TPs, we found the former group to be superior to the latter for all the previously mentioned leaf physiological traits. This superiority was related primarily to differences in chloroplast number and only secondarily to differences in activity and composition per chloroplast.     

Physiological and Biochemical Changes during Seed Filling in Relation to Leaf Senescence in Soybean

Field experiments with Glycine max (L.) Merr. cv. Ludou 11 and Ludou 4 were conducted to evaluate changes in photosynthetic rate, antioxidative enzyme activity, soluble protein, chlorophyll (Chl) and carotenoid (Car) contents in relation to leaf senescence during seed filling period. Photosynthetic rate, soluble protein content, catalase and peroxidase activities were the highest at 25 days after flowering (DAF). Chl a, Chl b and Car contents reached the maximum at 15 DAF and rapidly decreased after 33 DAF.