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1.
Crassulacean acid metabolism (CAM) is a specialized mode of photosynthesis that improves water use efficiency by shifting part or all of net atmospheric CO2 uptake to the night. Genetic dissection of regulatory and metabolic attributes of CAM has been limited by the difficulty of identifying a reliable phenotype for mutant screening. We developed a novel and simple colorimetric assay to measure leaf pH to screen fast neutron-mutagenized populations of common ice plant (Mesembryanthemum crystallinum), a facultative CAM species, to detect CAM-deficient mutants with limited nocturnal acidification. The isolated CAM-deficient mutants showed negligible net dark CO2 uptake compared with wild-type plants following the imposition of salinity stress. The mutants and wild-type plants accumulated nearly comparable levels of sodium in leaves, but the mutants grew more slowly than the wild-type plants. The mutants also had substantially reduced seed set and seed weight relative to wild type under salinity stress. Carbon-isotope ratios of seed collected from 4-month-old plants indicated that C3 photosynthesis made a greater contribution to seed production in mutants compared to wild type. The CAM-deficient mutants were deficient in leaf starch and lacked plastidic phosphoglucomutase, an enzyme critical for gluconeogenesis and starch formation, resulting in substrate limitation of nocturnal C4 acid formation. The restoration of nocturnal acidification by feeding detached leaves of salt-stressed mutants with glucose or sucrose supported this defect and served to illustrate the flexibility of CAM. The CAM-deficient mutants described here constitute important models for exploring regulatory features and metabolic consequences of CAM. 相似文献
2.
Summary In crassulacean acid metabolism (CAM) large amounts of malic acid are redistributed between vacuole and cytoplasm in the course
of night-to-day transitions. The corresponding changes of the cytoplasmic pH (pHcyt) were monitored in mesophyll protoplasts from the CAM plantKalanchoe daigremontiana Hamet et Perrier by ratiometric fluorimetry with the fluorescent dye 2′,7′-bis-(2-carboxyethyl)-5-(and-6-)carboxyfluorescein
as a pHcyt indicator. At the beginning of the light phase, pHcyt was slightly alkaline (about 7.5). It dropped during midday by about 0.3 pH units before recovering again in the late-day-to-early-dark
phase. In the physiological context the variation in pHcyt may be a component of CAM regulation. Due to its pH sensitivity, phosphoenolpyruvate carboxylase appears as a likely target
enzyme. From monitoring ΔpHcyt in response to loading the cytoplasm with the weak acid salt K-acetate a cytoplasmic H+-buffer capacity in the order of 65 mM H+ per pH unit was estimated at a pHcyt of about 7.5. With this value, an acid load of the cytoplasm by about 10 mM malic acid can be estimated as the cause of the
observed drop in pHcyt. A diurnal oscillation in pHcyt and a quantitatively similar cytoplasmic malic acid is predicted from an established mathematical model which allows simulation
of the CAM dynamics. The similarity of model predictions and experimental data supports the view put forward in this model
that a phase transition of the tonoplast is an essential functional element in CAM dynamics. 相似文献
3.
4.
Martin H. Spalding Geoffrey P. Arron Gerald E. Edwards 《Archives of biochemistry and biophysics》1980,199(2):448-456
Mitochondria isolated from the Crassulacean acid metabolism plant Sedum praealtum were demonstrated to decarboxylate added malate at basal rates of 30–50 μmol mg?1 original chlorophyll h?1. The basal rate could be stimulated markedly by the addition of ADP, oxaloacetic acid, an uncoupler of oxidative phosphorylation, or NAD, with maximum rates of 70–100 μmol mg?1 original chlorophyll h?1 observed. These observed rates were high enough to account for a large proportion of the estimated rate of malate decarboxylation in vivo. The major products of malate oxidation by the mitochondria in most cases were found to be pyruvate and CO2, indicating that malate oxidation in these mitochondria proceeds mainly through NAD malic enzyme rather than NAD malate dehydrogenase. Under conditions employed little of the pyruvate formed was further oxidized, suggesting a fate other than oxidation (conversion to starch) for this pyruvate. Malate decarboxylation by mitochondria and by partially purified NAD malic enzyme was markedly inhibited by NaHCO3. A possible physiological role is suggested for this inhibition as a feedback control on the enzyme. 相似文献
5.
Endogenous free-running regular circadian oscillations of net CO2 exchange in the crassulacean-acidmetabolism (CAM) plant Kalanchoë daigremontiana Hamet et Perrier de la Bâthie under constant external conditions in continuous light have been shown to change to irregular non-predictable (chaotic) time behaviour as irradiance or temperature are raised above a critical level. A model of CAM has been constructed with pools of major metabolites of varying concentrations, flows of metabolites leading to exchange between pools, metabolite transformations determined by chemical reactions, and feedback regulations. The model is described by a system of coupled non-linear differential equations. It shows stable rhythmicity in normal dark-light cycles and in continuous light and, like the K. daigremontiana leaves in the experiments, a change to chaos as irradiance is increased. The maintenance of endogenous oscillations in the model is brought about by a hysteresis switch or beat oscillator between two stable oscillation modes. In CAM these stable modes are vacuolar malate accumulation and remobilization. The model shows that the physical nature of the beat oscillator in the leaves can be explained by the balance between active and passive transport at the tonoplast.Abbreviations CAM
crassulacean acid metabolism
- D
dark period
- DL
12:12 h dark-light rhythm
- L
light period
- LL
constant illumination
- PPFD
photosynthetic photon flux density
- TL
leaf temperature
It is a great pleasure to thank Dr. G.-H. Vieweg, (Roßdorf, FRG) for his long-lasting efforts to have the phytotron in Darmstadt erected and for his persistent involvement during the various phases of planning and building. This made the present experiments possible. Dr. D. Kramer is thanked for all the time he spends to maintain functioning of the facility. Dr. P. Keller and Ms. Erika Ball assisted with the gas-exchange technology and helped with the surveillance of the long-running experiments, and Ms. Erika Ball performed all the integrations. Ms. Doris Schäfer is thanked for drawing the gas-exchange curves for publication. We are also most grateful to Professor Chr. Giersch and Professor M. Kluge (both Institut für Botanik, Technische Hochschule Darmstadt, FRG) for valuable discussions. 相似文献
6.
Abiotic drivers of environmental stress have been found to induce CAM expression (nocturnal carboxylation) in facultative CAM species such as Mesembryanthemum crystallinum. The role played by biotic factors such as competition with non-CAM species in affecting CAM expression, however, remains largely understudied. This research investigated the effects of salt and water conditions on the competition between M. crystallinum and the C3 grass Bromus mollis with which it is found to coexist in California’s coastal grasslands. We also investigated the extent to which CAM expression in M. crystallinum was affected by the intensity of the competition with B. mollis. We found that M. crystallinum had a competitive advantage over B. mollis in drought and saline conditions, while B. mollis exerted strong competitive effects on M. crystallinum in access to light and soil nutrients in high water conditions. This strong competitive effect even outweighed the favorable effects of salt or water additions in increasing the biomass and productivity of M. crystallinum in mixture. Regardless of salt conditions, M. crystallinum did not switch to CAM photosynthesis in response to this strong competitive effect from B. mollis. Disturbance (i.e., grass cutting) reduced the competitive pressure by B. mollis and allowed for CAM expression in M. crystallinum when it was grown mixed with B. mollis. We suggest that moderate competition with other functional groups can enhance CAM expression in M. crystallinum, thereby affecting its plasticity and ability to cope with biological stress. 相似文献
7.
The optimal conditions for opening of stomata in detached epidermis of the Crassulacean Acid Metabolism (CAM) plant Kalanchoe daigremontiana were determined. Stomatal opening in CO2–free air was unaffected by light so subsequently all epidermal strips were incubated in the dark and in CO2–free air. Apertures were maximal after 3 h incubation and were significantly greater at 15° C than 25° C. Thus stomata in isolated epidermis of this species can respond directly to temperature. Stomatal opening was greatest when the incubating buffer contained 17.6 mol m–3 K+, but decreased linearly with increasing K+ concentrations between 17.6 and 300 mol m–3; the decrease in aperture was shown to be associated with increasing osmotic potentials of the solutions. Reasons for this behaviour, which differs from that of many C3 and C4 species, are discussed. Stomatal apertures declined linearly upon incubation of epidermis on buffer solutions containing between 10–11 and 10–5 mol m–3 abscisic acid (ABA). Hence stomata on isolated epidermis of K. daigremontiana respond to lower concentrations of ABA than those of any species reported previously. 相似文献
8.
The role of acid phosphatases in plant phosphorus metabolism 总被引:18,自引:0,他引:18
Hydrolysis of phosphate esters is a critical process in the energy metabolism and metabolic regulation of plant cells. This review summarizes the characteristics and putative roles of plant acid phosphatase (APase). Although immunologically closely related, plant APases display remarkable heterogeneity with regards to their kinetic and molecular properties, and subcellular location. The secreted APases of roots and cell cultures are relatively non-specific enzymes that appear to be important in the hydrolysis and mobilization of Pi from extracellular phosphomonoesters for plant nutrition. Intracellular APases are undoubtedly involved in the routine utilization of Pi reserves or other Pi -containing compounds. A special class of intracellular APase exists that demonstrate a clear-cut (but generally nonabsolute) substrate selectivity. These APases are hypothesized to have distinct metabolic functions and include: phytase, phosphoglycolate phosphatase, 3-phosphoglycerate phosphatase, phosphoenolpyruvate phosphatase, and phosphotyrosyl-protein phosphatase. APase expression is regulated by a variety of developmental and environmental factors. Pi starvation induces de novo synthesis of extra- and intracellular APases in cell cultures as well as in whole plants. Recommendations are made to achieve uniformity in the analyses of the different APase isoforms normally encountered within and between different plant tissues. 相似文献
9.
The effects of root anoxia on a tobacco (Nicotiana tabacum) wild type (WT) and a transformant (LNR-H) lacking root nitrate reductase were compared. LNR-H plants were visibly more sensitive to oxygen deprivation than WT, showing rapid and heavy wilting symptoms. LNR-H roots also produced substantially more ethanol and lactate than WT roots under anoxia, and their sugar and sugar-P content, as well as their ATP levels, remained higher. The fermentation rates of WT and LNR-H roots were unaffected by sugar feeding and the higher fermentation rate in the LNR-H roots was associated with a greater acidification of the cytoplasm under anoxia. From these observations it is concluded: (i) that the absence of NR activity in the LNR-H roots does not necessarily limit NADH recycling; and (ii) that nitrate reduction in the WT roots results in a more acidifying metabolism. It is the higher metabolic rate in the LNR-H roots that leads to the greater cytoplasmic acidification under anoxia despite the absence of a contribution from the metabolism of nitrate. Competition for NADH cannot explain this difference in metabolic rate, and it remains unclear why the NR-free LNR-H, and tungstate-treated WT roots, had much higher fermentation rates than WT roots. The difference in anaerobic metabolism could still be due to the presence or absence of nitrate reductase and the possibility that this could occur through the production of nitric oxide is discussed. 相似文献
10.
The combination of a chamber for CO2 gas exchange with a potometric measuring arrangement allowed concomitant investigations into CO2 gas exchange, transpiration and water uptake by the roots of whole plants of Senecio medley-woodii, a species which exhibits Crassulacean acid metabolism. The water-uptake rate showed the same daily pattern as malate concentration and osmotic potential. The accumulation of organic acids resulting from nocturnal CO2 fixation enhanced the water-uptake rate from dusk to dawn. During the day the water-uptake rates decreased with decreasing organic-acid concentration. With gradually increasing water stress, CO2 dark fixation of S. medley-woodii was increased as long as water could be taken up by the roots. It was also shown that a reestablished water supply after drought caused a similar increase which in both cases ameliorated the water uptake in order to conserve a positive water balance for as long as possible. This water-uptake pattern shows that Crassulacean acid metabolism is not only a water-saving adaptation but also enhances water uptake and is directly correlated with the amelioration of the plant water status.Abbreviation CAM
Crassulacean acid metabolism 相似文献
11.
12.
Proteases play important roles in plant innate immunity. In this mini-review, we describe the current view on the role of a plant protease, vacuolar processing enzyme (VPE), and the first identified plant caspase-1-like protein, in plant immunity. In the past several years, VPEs were determined to play important roles in various types of cell death in plants. Early studies demonstrated the identification of VPE as a vacuolar hydrolytic protein responsible for maturation of vacuolar proteins. Later, Nicotiana benthamiana VPE was reported to mediate virus-induced hypersensitive response by regulating membrane collapse. The ortholog of VPE in Arabidopsis is also suggested to be involved in both mycotoxin-induced cell death and developmental cell death. However, the role of VPE in elicitor-signaling is still unclear. Our recent studies demonstrated the involvement of VPE in elicitor signal transduction to induce stomatal closure and defense responses, including defense gene expression and hypersensitive cell death.Key words: hypersensitive cell death, elicitor, stomatal closure, pathogen-associated molecular patterns, plant innate immunity, programmed cell deathIn the course of their development, plants have had to face a wide range of potential pathogens, including viral, bacterial, fungal and oomycete pathogens. Plants, unlike animals, which have specialized defender cells and an adaptive immune system, have an innate immunity of each cell and produce systemic signals emanating from the infection site. The plant innate immunity (PTI) is induced by pathogen-associated molecular patterns (PAMPs)1 and elicitors.2,3 However, some pathogens deliver virulence proteins that target host protein to overcome the plant immunity response. Most plants have evolved the corresponding resistance (R) protein to recognize effector activity, which will trigger plant resistance through effector-triggered immunity (ETI).4 Natural selection drives evolution of new pathogen effector proteins and plant R proteins. This tug-of-war between plants and pathogens is represented as a zig-zag-zig model.5–7 Both PTI and ETI cause stomatal closure and hypersensitive response (HR), a programmed host cell death (PCD) to limit pathogen development.5,8 In plants, HR is caused by proteases with caspase activity. At least eight caspase activities have now been measured in plant extracts, which were found using caspase substrates, and various caspase inhibitors can block many forms of plant programmed cell death.9In the past several years, vacuolar-processing enzyme (VPE) has been determined to play important roles in plant immunity responses. In this review paper, I describe the current view on the role of VPE in plant immunity, based on our own research and recent developments in this field. 相似文献
13.
J T Brosnan 《Canadian journal of physiology and pharmacology》1987,65(12):2355-2362
Measurement of the arteriovenous differences for free amino acids across rat kidney reveals that glycine and citrulline are removed and serine and arginine are added to the circulation. In addition, glutamine is taken up in large quantities by kidneys of animals that need to excrete large quantities of acid (e.g., diabetic animals, NH4Cl-fed animals, and animals fed a high protein diet). Glutamine is the major precursor of urinary ammonia and thus renal glutamine metabolism plays a key role in acid-base homeostasis. This process occurs primarily in the cells of the convoluted proximal tubule. Glutamine carbon is converted to glucose in acidotic rats and is totally oxidized in dogs. Regulation of glutamine metabolism occurs at two levels: acute regulation and chronic regulation. Acute regulation is, in part, mediated through a fall in intracellular [H+]. This activates alpha-ketoglutarate dehydrogenase and, ultimately, glutaminase. Chronic regulation involves induction of key enzymes, including, in the rat, glutaminase, glutamate dehydrogenase, and phosphoenolpyruvate carboxykinase. During the acidosis of prolonged starvation, the kidneys' requirement for glutamine must be met from muscle proteolysis and thus becomes a drain on lean body mass. Serine synthesis occurs by two separate pathways: from glycine by the combined actions of the glycine cleavage enzyme and serine hydroxymethyltransferase and from gluconeogenic precursors using the phosphorylated-intermediate pathway. Both pathways are located in the cells of the proximal tubule. Conversion of glycine to serine is ammoniagenic and the activity of the glycine cleavage enzyme is increased in acidosis. The function of serine synthesis by the phosphorylated-intermediate pathway is not apparent.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
14.
Leaves of the Crassulacean acid metabolism (CAM) plant Kalancho? daigremontiana Hamet et Perrier de la Bathie show overt circadian rhythms in net CO2 uptake, leaf conductance to water and intercellular CO2 concentration, which are entrained by periodic temperature cycles. To probe their sensitivity to thermoperiodic perturbations, intact leaves were exposed to continuous light intensity and temperature cycles with a period of 16 h, applying a set of different baseline temperatures and thermodriver amplitudes. All three overt rhythms were analyzed with respect to their frequency spectra and their phase relations with the thermodriver. For most stimulation protocols, stomatal conductance and net CO2 change were fully or partially entrained by the temperature pulses, while the internal CO2 concentration remained dominated by oscillations in the circadian range. Prolonged time series recorded for up to 22 d in continuous light underline the robustness of these circadian oscillations. This suggests that the overt circadian rhythm of net CO2 uptake in CAM results from the interaction of two coupled original systems: (i) an endogenous cycle of CO2 fixation in the mesophyll, showing very robust periodic activity, and (ii) stomatal movements that respond to environmental stimuli independently of rhythmic processes in the mesophyll, and thus modulate the gas exchange amplitude. 相似文献
15.
Having gained some understanding of the consequences of the CO(2)-concentrating mechanisms in crassulacean acid metabolism (CAM) that internalize the photosynthetic environment of the Cretaceous on a daily basis, it may be time to consider potential long-term effects of the planetary CO(2)-concentrating mechanism on growth and ecology of these plants in the Anthropocene. This paper emphasizes our limited understanding of the carbohydrate economy of CAM in relation to growth processes and briefly reviews recent studies of the diel cycles of growth in these plants. An inadvertent long-term, regional-scale experiment from the past is revisited in which an Opuntia monoculture grew to occupy >25 million hectares of farmland in central eastern Australia, producing a total biomass of about 1.5 billion tonnes in about 80 years. Although at the time it does not seem to have been recognized that this invasion involved CAM, a botanist from the University of Melbourne, Jean White-Haney emerges as a heroic pioneer in the control of the invader by poison and pioneered its biological control. The Opuntia population was expanding at 10-100 ha h(-1) when it was brought to a halt within a decade by the voracious appetite of Cactoblastis cactorum larvae. It is now known that the female parent moth of this predator detects CAM in O. stricta prior to oviposition by deploying the most sensitive CO(2) detector system yet found in the Lepidoptera. The O. stricta invasion is a dramatic demonstration of the capacity of CAM plants to attain and sustain high biomass; to sequester and retain atmospheric CO(2). In conclusion, experiments are reviewed that show stimulation of CO(2) assimilation, growth, and biomass of CAM plants by elevated atmospheric [CO(2)], and the proposition that these plants may have a role in atmospheric CO(2) sequestration is re-examined. This role may be compromised by predators such as Cactoblastis. However the moth CO(2) sensors are adapted to pre-industrial atmospheric [CO(2)] and FACE (free-air CO(2) enrichment) experiments show this exquisite system of biological control is also compromised by rising global [CO(2)] in the Anthropocene. 相似文献
16.
17.
Two tobacco lines with (Nicotiana tabacum cv. Gatersleben, WT) or without (transformant LNR-H) nitrate reductase in roots were chosen as model systems to re-evaluate the role of root nitrate reduction for survival of anoxia. In this first paper, the two hydroponically grown lines were compared with respect to their root morphology, root respiration and the root content of inorganic cations, anions, and metabolites. Leaf transpiration in relation to root morphology was also determined. In comparison to WT roots containing NR, the NR-free LNR-H transformants had slightly shorter and thicker roots with a lower root surface area per g leaf FW. Consistent with that, LNR-H leaves had lower transpiration rates than WT. LNR-H-roots also showed consistently higher respiration and higher contents of ATP, starch and hexose monophosphates than WT roots. Concentrations of free sugars were only slightly higher in LNR-H roots. Total soluble protein content was identical in both lines, whereas amino acids were higher in LNR-H. Contents of major inorganic cations and anions were also almost identical in both lines. We conclude that WT versus LNR-H plants are a suitable tool to re-evaluate the role of nitrate reduction in flooding tolerance. 相似文献
18.
Guralnick LJ Ku MS Edwards GE Strand D Hockema B Earnest J 《Plant & cell physiology》2001,42(2):236-239
The induction of Crassulacean acid metabolism in M:esembryanthemum crystallinum was investigated in response to foliar application of gibberellic acid (GA). After 5 weeks of treatment, GA-treated plants showed 1.7- to almost a 4-fold increase of phosphoenolpyruvate carboxylase (PEPcase) activity with a concomitant increase in acid metabolism when compared to control plants. Immunoblot analysis indicated an increase in the PEPcase protein similar to that of salt treatment while Rubisco did not show a similar rise. The results indicate that exogenously applied GA accelerates plant developmental expression of PEPcase and Crassulacean acid metabolism in M: crystallinum. 相似文献
19.
《Plant Physiology and Biochemistry》2003,41(2):97-105
An NAD-malate dehydrogenase (NAD-MDH, EC 1.1.1.37) was purified and characterized from leaves of Aptenia cordifolia L. f. (Schwant). This plant performs crassulacean acid metabolism (CAM), as indicated by: (a) elevated levels of phosphoenolpyruvate carboxylase (PEPC) and NAD(P) malic enzyme; (b) regulation of PEPC compatible with its function during the night; (c) characteristic day/night changes in titratable acidity; and (d) gas exchange profile consistent with that shown by CAM plants. These features remained unchanged by water availability or salt stress, suggesting constitutive CAM. The purified MDH showed a subunit molecular mass of 39.4 kDa, a native mass of 83 kDa (dimer) and a pI of 5.8. It cross-reacted with antibodies against cytosolic malate dehydrogenase (cMDH) from pineapple. Maximum activities for oxaloacetate (OAA) reduction or malate oxidation were observed at pH 7.0 and between pH 7.2 and 8.4, respectively. The enzyme was inhibited by excess OAA, in a pH-dependent manner. A discontinuity was observed in Arrhenius plots at 33 °C, with an activation energy twice as high below this temperature. Although immunologically related, some physical and kinetic dissimilarities between the A. cordifolia and pineapple enzymes suggest that diverse CAM metabolic subtypes may require different MDH isozymes to carry out OAA reduction. 相似文献
20.
M. Fernanda Ricalde José Luis Andrade Rafael Durán Juan Manuel Dupuy J. Luis Simá Roberth Us-Santamaría Louis S. Santiago 《Oecologia》2010,164(4):871-880
Expression of crassulacean acid metabolism (CAM) is characterized by extreme variability within and between taxa and its sensitivity
to environmental variation. In this study, we determined seasonal fluctuations in CAM photosynthesis with measurements of
nocturnal tissue acidification and carbon isotopic composition (δ13C) of bulk tissue and extracted sugars in three plant communities along a precipitation gradient (500, 700, and 1,000 mm year−1) on the Yucatan Peninsula. We also related the degree of CAM to light habitat and relative abundance of species in the three
sites. For all species, the greatest tissue acid accumulation occurred during the rainy season. In the 500 mm site, tissue
acidification was greater for the species growing at 30% of daily total photon flux density (PFD) than species growing at
80% PFD. Whereas in the two wetter sites, the species growing at 80% total PFD had greater tissue acidification. All species
had values of bulk tissue δ13C less negative than −20‰, indicating strong CAM activity. The bulk tissue δ13C values in plants from the 500 mm site were 2‰ less negative than in plants from the wetter sites, and the only species growing
in the three communities, Acanthocereus tetragonus (Cactaceae), showed a significant negative relationship between both bulk tissue and sugar δ13C values and annual rainfall, consistent with greater CO2 assimilation through the CAM pathway with decreasing water availability. Overall, variation in the use of CAM photosynthesis
was related to water and light availability and CAM appeared to be more ecologically important in the tropical dry forests
than in the coastal dune. 相似文献