Senescence of Rice Leaves VI. Comparative Study of the Metabolic Changes of Senescing Turgid and Water-Stressed Excised leaves

Some metabolic changes of senescing turgid and water-stressedexcised rice leaves were compared under incubation in the dark.The decrease of the chlorophyll and protein level and the increaseof the -amino nitrogen were faster in the water-stressed leavesthan in the turgid leaves during the first two or three daysof incubation. However, the changes in levels of chlorophyll,protein and a-amino nitrogen were later retarded by water stress.The rate of decline in soluble sugar was more rapid in the turgidthan in the water-stressed leaves. In turgid leaves, there wasa slight but significant increase in the proline content inthe first day of incubation; subsequently, proline accumulatedrelatively rapidly, yet at a slower rate than in the stressedleaves. The activity of acid inorganic pyrophosphatase increased,but that of alkaline inorganic pyrophosphatase decreased duringthe senescence of both turgid and water-stressed leaves. Theenzyme activities are, therefore, due to two separate enzymeproteins. Water stress enhanced the increase of acid inorganicpyrophosphatase activity only during the first two days; itretarded the decrease of alkaline inorganic pyrophosphataseactivity at later stage of incubation. It is concluded that water stress does not accelerate all themetabolic changes associated with the senescence of excisedleaves. (Received January 24, 1981; Accepted March 27, 1981)     

Nitrogen-deficiency-induced loss in photosynthesis and modulation of β-galactosidase activity during senescence of <Emphasis Type="Italic">Arabidopsis</Emphasis> leaves

Senescence-induced loss in the content of chlorophyll and the rate of oxygen evolution is remarkably enhanced when the leaves of Arabidopsis thaliana experience nitrogen-deficiency stress. On the other hand, the decline in the level of total soluble sugar during senescence is very slow and nitrogen deficiency does not exhibit any further change. The relative stability in the level of the sugar in the background of severe decline of photosynthesis may suggest the contribution of sugars from other sources to sustain its homeostasis to execute and complete energy-dependent senescence process and stress response. The possible participation of cell wall polysaccharides contributing to sugar homeostasis is predicted. Senescence-induced increase in the activity of β-galactosidase (EC 3.2.1.23) and its further enhancement in senescing leaves experiencing nitrogen stress support the proposition of participation of the enzyme for breakdown of the wall polysaccharides to sugars. The loss of photosynthesis as a possible signal for enhancement in the activity of β-galactosidase has been further examined in the excised leaves incubated in Okada and Shimura (OS) nutrient medium with and without nitrogen. Nitrogen limitation experienced by excised leaves causes rapid loss in photosynthesis with concomitant increase in the activity of the enzyme extracted both from soluble and cell wall fractions. The differential activity of the enzyme from soluble and cell wall fractions during development-dependent leaf senescence and premature senescence in excised leaves induced by nitrogen deficiency appears to be complex and needs to be resolved in the future.     

Comparative Study of the Metabolism of 1-Aminocyclopropane-1-carboxylic Acid and Senescence of Water-Stressed and ABA-Treated Excised Rice Leaves

Changes in the metabolism of 1-aminocyclopropane-l-carboxylicacid (ACC) during senescence in the light in turgid, water-stressed,and ABA-treated, excised rice leaves were examined. The decreasesin levels of Chl and protein were more rapid in the water-stressedand in the ABA-treated leaves than in the turgid leaves. Inturgid leaves, levels of proline remained very low, but theyincreased considerably as a result of water stress or treatmentwith ABA. The production of ethylene was strongly inhibitedby water stress and by ABA through the inhibition of the synthesisof ACC and/or the conversion of ACC to ethylene. In turgid leaves,the level of 1-(malonylamino)cyclopropane-l-carboxylic acid(MACC) increased with time during incubation in the light. Waterstress resulted in a pattern of accumulation of MACC similarto that in the turgid control. However, ABA blocked the malonylationof ACC. (Received July 27, 1989; Accepted March 12, 1990)     

Uniconazole-induced tolerance of soybean to water deficit stress in relation to changes in photosynthesis, hormones and antioxidant system

This study investigated whether uniconazole confers drought tolerance to soybean and if such tolerance is correlated with changes in photosynthesis, hormones and antioxidant system of leaves. Soybean plants were foliar treated with uniconazole at 50 mg L-1 at the beginning of bloom and then exposed to water deficit stress at pod initiation for 7 d. Uniconazole promoted biomass accumulation and seed yield under both water conditions. Plants treated with uniconazole showed higher leaf water potential only in water-stressed condition. Water stress decreased the chlorophyll content and photosynthetic rate, but those of uniconazole-treated plants were higher than the stressed control. Uniconazole increased the maximum quantum yield of photosystemand ribulose-1,5-bisphosphate carboxylase/oxygenase activity of water-stressed plants. Water stress decreased partitioning of assimilated 14C from labeled leaf to the other parts of the plant. In contrast, uniconazole enhanced translocation of assimilated 14C from labeled leaves to the other parts, except stems, regardless of water treatment. Uniconazole-treated plants contained less GA3, GA4 and ABA under well-watered condition than untreated plants, while the IAA and zeatin levels were increased substantially under both water conditions, and ABA concentration was also increased under water stressed condition. Under water-stressed conditions, uniconazole increased the content of proline and soluble sugars, and the activities of superoxide dismutase and peroxidase in soybean leaves but not the malondialdehyde content or electrical conductivity. These results suggest that uniconazole-induced tolerance to water deficit stress in soybean was related to the changes of photosynthesis, hormones and antioxidant system of leaves.     

Drought-induced male sterility in rice: Changes in carbohydrate levels and enzyme activities associated with the inhibition of starch accumulation in pollen

Male reproductive development of rice (Oryza sativa L.) is very sensitive to drought. A brief, transitory episode of water stress during meiosis in pollen mother cells of rice grown under controlled environmental conditions induced pollen sterility. Anthers containing sterile pollen were smaller, thinner, and often deformed compared to normal anthers of well-watered plants. Only about 20% of the fully developed florets in stressed plants produced grains, compared to 90% in well-watered controls. Water stress treatments after meiosis were progressively less damaging. Levels of starch and sugars and activities of key enzymes involved in sucrose cleavage and starch synthesis were analyzed in anthers collected at various developmental stages from plants briefly stressed during meiosis and then re-watered. Normal starch accumulation during pollen development was strongly inhibited in stress-affected anthers. During the period of stress, both reducing and non-reducing sugars accumulated in anthers. After the relief of stress, reducing sugar levels fell somewhat below those in controls, but levels of non-reducing sugars remained higher than in controls. Activities of acid invertase and soluble starch synthase in stressed anthers were lower than in controls at comparable stages throughout development, during as well as after stress. Stress had no immediate effect on ADP-glucose pyrophosphorylase activity, but had an inhibitory aftereffect throughout post-stress development. Sucrose synthase activity, which was, relatively speaking, much lower than acid invertase activity, was only slightly suppressed by stress. The results show that it is unlikely that pollen sterility, or the attendant inhibition of starch accumulation, in water-stressed rice plants are caused by carbohydrate starvation per se. Instead, an impairment of enzymes of sugar metabolism and starch synthesis may be among the potential causes of this failure.     

Expression of E. coli inorganic pyrophosphatase in transgenic plants alters photoassimilate partitioning

Transgenic plants were constructed expressing a novel cytosolic inorganic pyrophosphatase in order to reduce the cytosolic pyrophosphate content. To this end the Escherichia coli gene ppa encoding inorganic pyrophosphatase was cloned between the 35S CaMV promoter and the poly(A) site of the octopine synthase gene and transferred into tobacco and potato plants by Agrobacterium-mediated gene transfer. Regenerated plants were tested for the expression of the ppa gene by Northern blots and activity gels. Plants expressing active inorganic pyrophosphatase showed a dramatic change in photoassimilate partitioning. In both transgenic tobacco and potato plants the ratio between soluble sugars and starch was increased by about 3-4-fold in source leaves as compared with the wild-type. However, whereas source leaves of transgenic tobacco plants accumulated much higher levels of glucose (up to 68-fold), fructose (up to 24-fold), sucrose (up to 12-fold) and starch (up to 8-fold) this was not observed in potato plants where the change in assimilate partitioning in source leaves was due to an increase of about 2-fold in sucrose and a reduction in starch content. Expression of the cytosolic inorganic pyrophosphatase in tobacco results in stunted growth of vegetatively growing plants due to a reduced internode distance. Upon flowering the transgenic plants increase their growth rate, reaching almost the same height as control plants at the end of the growth period. Old source leaves accumulate up to 100-fold more soluble sugars than control leaves. This increase in soluble sugars is accompanied by a reduction in chlorophyll content (up to 85%). Transgenic potato plants showed a less dramatic change in their growth behaviour. Plants were slightly reduced in size, with stems more highly branched. Tuber number increased 2-3-fold, but tuber weight was lower resulting in no net increase in fresh weight.     

Pyrophosphatase, Peroxidase and Polyphenoloxidase Activities during Leaf Development and Senescence

Inorganic pyrophosphatase, peroxidase, and polyphenoloxidase activities were studied as the function of leaf insertion level in eight monocotyledonous and eight dicotyledonous species. Alkaline inorganic pyrophosphatase shows a declining activity toward the end of senescence whereas no regular drift in either peroxidase or polyphenoloxidase activities was noticed during senescence of attached leaves. In the primary leaves of rice, peroxidase and polyphenoloxidase activities were high in the senescent leaves and there exists a correlation between chlorophyll content and peroxidase activity though not with polyphenoloxidase activity. Upon detachment leaves exhibit increasing peroxidase and polyphenoloxidase activities with time. The distribution of the enzyme activities during senescence of attached leaves is suggested to be species-specific, and an increase in peroxidase and polyphenoloxidase activities cannot be taken as an indicator of leaf senescence.     

Induction of leaf senescence by low nitrogen nutrition in sunflower (Helianthus annuus) plants

Different parameters which vary during the leaf development in sunflower plants grown with nitrate (2 or 20 mM) for a 42‐day period have been determined. The plants grown with 20 mM nitrate (N+) showed greater leaf area and specific leaf mass than the plants grown with 2 mM nitrate (N?). The total chlorophyll content decreased with leaf senescence, like the photosynthetic rate. This decline of photosynthetic activity was greater in plants grown with low nitrogen level (N?), showing more pronounced senescence symptoms than with high nitrogen (N+). In both treatments, soluble sugars increased with aging, while starch content decreased. A significant increase of hexose to sucrose ratio was observed at the beginning of senescence, and this raise was higher in N? plants than in N+ plants. These results show that sugar senescence regulation is dependent on nitrogen, supporting the hypothesis that leaf senescence is regulated by the C/N balance. In N+ and N? plants, ammonium and free amino acid concentrations were high in young leaves and decreased progressively in the senescent leaves. In both treatments, asparagine, and in a lower extent glutamine, increased after senescence start. The drop in the (Glu+Asp)/(Gln+Asn) ratio associated with the leaf development level suggests a greater nitrogen mobilization. Besides, the decline in this ratio occurred earlier and more rapidly in N? plants than in N+ plants, suggesting that the N? remobilization rate correlates with leaf senescence severity. In both N+ and N? plants, an important oxidative stress was generated in vivo during sunflower leaf senescence, as revealed by lipid peroxidation and hydrogen peroxide accumulation. In senescent leaves, the increase in hydrogen peroxide levels occurred in parallel with a decline in the activity of antioxidant enzymes. In N+ plants, the activities of catalase and ascorbate peroxidase (APX) increased to reach their highest values at 28 days, and later decreased during senescence, whereas in N? plants these activities started to decrease earlier, APX after 16 days and catalase after 22 days, suggesting that senescence is accelerated in N‐leaves. It is probable that systemic signals, such as a deficit in amino acids or other metabolites associated with the nitrogen metabolism produced in plants grown with low nitrogen, lead to an early senescence and a higher oxidation state of the cells of these plant leaves.     

Ecophysiological adaptations of Asphodelus aestivus to Mediterranean climate periodicity: water relations and energetic status

During the course of a year, we studied the water and metabolic status of Asphodelus aestivus , a native geophyte of East Mediterranean, dominant in areas degraded by overgrazing and fire The species proved to be very efficient m storing water during the long summer drought At soil water content values around zero, in the upper part of the soil profile (10–20 cm in depth), the roots remained hydrated and turgid, their relative water content was > 60% and water potential > - 1 6 MPa Accumulation of proline during winter in leaves (ca 5 mg g^–1 dry weight) and tubers (though at significantly lower levels, ca 1.5 mg g^–1) might be taken as evidence of a winter cold stress response Proline accumulation in tubers, under summer drought, was similar to that in winter Maximal values of caloric content were recorded in expanding leaves (ca 5600 cal g^–1 dry weight) and minimal before leaf senescence In contrast, root caloric content remained fairly constant for most of the year (ca 4550 cal g^–1 dry weight) in spite of drastic changes in the concentrations of soluble sugars, starch and lipids Long before senescence, photosynthetic products were translocated to the below ground system, where they were stored Drastic changes of the storage compounds were observed before emergence of the flowering stalk, far greater in magnitude than those before leaf emergence The below ground part of the species was found to be less susceptible to climatic stress and to constitute an energetically rather stable system The physiological processes of the species were well synchronised to the fluctuations of the Mediterranean climate     

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.     

Metabolic regulation of leaf senescence: interactions of sugar signalling with biotic and abiotic stress responses

Sugars are important signals in the regulation of plant metabolism and development. During stress and in senescing leaves, sugars often accumulate. In addition, both sugar accumulation and stress can induce leaf senescence. Infection by bacterial and fungal pathogens and attack by herbivores and gall-forming insects may influence leaf senescence via modulation of the sugar status, either by directly affecting primary carbon metabolism or by regulating steady state levels of plant hormones. Many types of biotic interactions involve the induction of extracellular invertase as the key enzyme of an apoplasmic phloem unloading pathway, resulting in a sourcesink transition and an increased hexose/sucrose ratio. Induction of the levels of the phytohormones ethylene and jasmonate in biotic interactions results in accelerated senescence, whereas an increase in plant- or pathogen-derived cytokinins delays senescence and results in the formation of green islands within senescing leaves. Interactions between sugar and hormone signalling also play a role in response to abiotic stress. For example, interactions between sugar and abscisic acid (ABA) signalling may be responsible for the induction of senescence during drought stress. Cold treatment, on the other hand, can result in delayed senescence, despite sugar and ABA accumulation. Moreover, natural variation can be found in senescence regulation by sugars and in response to stress: in response to drought stress, both drought escape and dehydration avoidance strategies have been described in different Arabidopsis accessions. The regulation of senescence by sugars may be key to these different strategies in response to stress.     

Seasonal changes in carbohydrates of perennial root nodules of beach pea

Seasonal changes in amyloplast, starch, total soluble sugars, non-reducing sugars and reducing sugars of perennial root nodules of beach pea (Lathyrus maritimus) were studied. Ultrastructural changes in nodule cells of beach pea indicated an accumulation of large amounts of amyloplasts with multiple starch grains in summer months. As the winter season sets in, degradation of amyloplasts and starch grains was detected. The membranes of amyloplasts faded out in winter and the structure of the amyloplasts was lost. The degradation of starch grains showed some electron-dense fiber-like material and amorphous structures. Quantitative data revealed an increase in starch reserves during the summer and depletion during the winter. Total soluble sugar and non-reducing sugar concentrations peaked in the middle of the winter, whereas reducing sugar concentrations showed an increase in the fall. These results indicate that elevated levels of various sugars most likely help to maintain high osmolarity of cells so that the dormant nodules do not freeze during the prolonged periods of cold in the winter.     

Inhibition of proline oxidation by water stress

The conversion of proline to glutamic acid and hence to other soluble compounds (proline oxidation) proceeds readily in turgid barley (Hordeum vulgare) leaves and is stimulated by higher concentrations of proline. This suggests that proline oxidation could function as a control mechanism for maintaining low cellular levels of proline in turgid tissue. In water-stressed tissue, however, proline oxidation is reduced to negligible rates. These results are consistent with the idea that proline accumulation results from inactivation by water stress of normal control mechanisms. It seems likely that inhibition of proline oxidation is necessary in maintaining the high levels of proline found in stressed barley leaves.     

蒙古莸幼苗干旱致死过程中非结构性碳水化合物的变化

以一年生蒙古莸幼苗为对象,设置适宜水分、慢速干旱致死和快速干旱致死3个处理,研究不同干旱强度致死下蒙古莸幼苗各器官中非结构性碳水化合物(NSC,包括可溶性糖和淀粉)的含量变化及其分配规律.结果表明:慢速干旱致死胁迫下各器官可溶性糖含量与适宜水分组无显著差异.随时间的推移,茎可溶性糖含量先增加后减少,淀粉和NSC含量增加;粗根可溶性糖含量减少,淀粉和NSC含量增加;叶可溶性糖含量增加,淀粉和NSC含量减少.致死时(80 d),叶、茎、粗根和细根的NSC含量分别为6.2%、7.8%、8.3%和7.4%.快速干旱致死胁迫下,各器官可溶性糖含量均高于适宜水分处理组,而淀粉和NSC含量均低于适宜水分组.随时间的推移,根可溶性糖含量下降,淀粉和NSC含量上升;茎可溶性糖、淀粉和NSC含量均上升;叶可溶性糖含量上升,淀粉和NSC含量下降.致死时(30 d),叶、茎、粗根和细根的NSC含量分别为5.9%、6.6%、8.9%和7.7%.应对不同的干旱致死情况,蒙古莸幼苗各器官间非结构性碳水化合物呈现出不同的动态变化.在慢速干旱致死胁迫下,NSC优先为维持各器官生理代谢活动提供能量;而在快速干旱致死下,NSC主要以可溶性糖形式维持植物代谢,调节渗透势,促进吸水,应对急剧的干旱胁迫.     

The Effect of Benzimidazole and Red Light on the Senescence of Detached Leaves of Oryza sativa cv. Ratna

Excised rice leaves (Oryza sativa L. cv. Ratna) werefloated on a 10^–3M solution of benzirnidazole under dark or continuous red light. Compared to the water control a degradation of chlorophyll, protein, RNA, DNA and a decrease in the activity of alkaline inorganic pyrophosphatase was delayed at the same time as an increase of α-amino nitrogen and the activity of acid inorganic pyrophosphatase occurred, Benzimidazole was more effective under red light than in the dark in retarding senescence. The possible role of inorganic pyrophosphatases is discussed with respect to biosynthesis during leaf senescence.     

Alteration of the Amount of the Chloroplast Phosphate Translocator in Transgenic Tobacco Affects the Distribution of Assimilate between Starch and Sugar

Tobacco plants (Nicotiana tabacum L.) transformed with sense and antisense constructs of a cDNA encoding the tobacco phosphate-triose phosphate-3-phosphoglycerate translocator (phosphate translocator) were shown to contain altered amounts of phosphate translocator mRNA and protein. Phosphate translocator activity in intact chloroplasts isolated from transformed plants showed a 15-fold variation, from 20% of the wild-type activity in antisense transformants to 300% of the wild-type activity in sense transformants. However, the maximal rates of photosynthesis and the rates of photosynthetic carbon assimilation in ambient CO2 showed no consistent differences between transformants. Starch content was decreased by 20% and total soluble sugars were increased by 20% in leaves of antisense transformants compared to sense transformants. The 40% decrease in the ratio of starch to total soluble sugars in antisense transformants relative to sense transformants indicates that distribution of assimilate between starch and sugar had been altered. However, the amount of sucrose in the leaves was unchanged. The changes in total soluble sugars were accounted for completely by changes in glucose and fructose, suggesting the existence of a homeostatic mechanism for maintaining sucrose concentrations in the leaves at the expense of glucose and fructose.     

Effects of Kinetin on Transpiration Rate and Abscisic Acid Content of Water Stressed Young Wheat Plants

Effects of kinetin on transpiration rate and abscisic acid content were determined. Leaves from 9-day-old wheat plants (Triticum aestivum L. cv. Weibull's Starke II) were used. —Transpiration rate decreased in excised leaves put in water, but it was maintained at a higher rate when kinetin was supplied. When excised leaves were water stressed by air-drying for 1 h, addition of kinetin resulted in a considerable stimulation of transpiration rate. The effect reached its maximum after 15 h and this level remained relatively unchanged for at least 10 h. Intact seedlings which were stressed before leaf excision, showed only a slight stimulation of kinetin on transpiration rate. — Abscisic acid content slowly increased up to three-fold in 2 days in excised leaves put in water. In excised and water-stressed leaves the abscisic acid content was reduced during the first 24 h and then increased. As the leaves were fully turgid, the increase could not have been caused by water stress. However, both in stressed and unstressed leaves kinetin addition reduced the increase in abscisic acid content. — It is suggested that the stimulation by kinetin on transpiration rate in excised and water stressed leaves was mainly due to the combined effect of (1) a reduction in the activity of endogenous cytokinins, (2) kinetin acting as a ‘substitute’ for the inactivated cytokinins but exerting a stronger effect on transpiration than the endogenous cytokinins, and (3) the ‘extra’ reduction in abscisic acid content caused by the kinetin treatment. Furthermore, the results indicate that changes in cytokinins might be partly responsible for the aftereffect on transpiration.     

Leaf senescence in response to elevated atmospheric CO<Subscript>2</Subscript> concentration and low nitrogen supply

This review reports the physiological and metabolic changes in plants during development under elevated atmospheric carbon dioxide concentration and/or limited-nitrogen supply in order to establish their effects on leaf senescence induction. Elevated CO₂ concentration and nitrogen supply modify gene expression, protein content and composition, various aspects of photosynthesis, sugar metabolism, nitrogen metabolism, and redox state in plants. Elevated CO₂ usually causes sugar accumulation and decreased nitrogen content in plant leaves, leading to imbalanced C/N ratio in mature leaves, which is one of the main factors behind premature senescence in leaves. Elevated CO₂ and low nitrogen decrease activities of some antioxidant enzymes and thus increase H₂O₂ production. These changes lead to oxidative stress that results in the degradation of photosynthetic pigments and eventually induce senescence. However, this accelerated leaf senescence under conditions of elevated CO₂ and limited nitrogen can mobilize nutrients to growing organs and thus ensure their functionality.