Physiological Responses of Zea mays Seedlings to Interactions Between Cadmium and Salinity

The effects of 0, 2.5, 5.0, and 10.0 pmol/L Cd^2＋（Cd（NO3）2.4H2O） and 0, 10, 25, 50, and 100 mmol/L NaCl on growth, photosynthesis and the content of some ions in maize （Zea mays L.） were investigated in the present study. With Increasing concentrations of Cd^2＋ or NaCI alone in Hoagland nutrient solution, the chlorophylls and starch content decreased. Combination treatment with salinity and cadmium increased the negative effects observed following the two stresses alone. Plants exhibiting growth retardation in response to one mild stress factor （25-50 mmol/L NaCl） became more tolerant to the other stress factor （Cd）. The exposure of plants to cadmium caused a partial reversal of the effects of salinity. Root and shoot growth, ion accumulation and levels of photosynthetic pigments were improved at moderate concentrations of the two stress factors Imposed jointly.     

Different Effects of Homocysteine and Oxidized Low Density Lipoprotein on Methylation Status in the Promoter Region of the Estrogen Receptor α Gene

We investigated the effects of homocysteine (Hcy) and oxidized low density lipoprotein (ox-LDL) on DNA methylation in the promoter region of the estrogen receptor α (ERos) gene,and its potentialmechanism in the pathogenesis of atherosclerosis.Cultured smooth muscle cells (SMCs) of humans weretreated by Hcy and ox-LDL with different concentrations for different periods of time.The DNA methylationstatus was assayed by nested methylation-specific polymerase chain reaction,the lipids that accumulated inthe SMCs and foam cell formations were examined with Oil red O staining.The proliferation of SMCs wasassayed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.The results showedthat ox-LDL in moderate concentrations (10-40 mg/L) induced de novo methylation in the promoter regionof the ERα gene of SMCs.However,high concentrations (50 mg/L) of ox-LDL,resulted in demethylation ofERα.The Hcy treatment resulted in de novo methylation in the promoter region of the ERα gene with aconcentration- and treating time-dependent manner,and a dose-dependent promoting effect on SMCproliferation.These data indicated that the two risk factors for atherosclerosis had the function of inducingde novo methylation in the promoter region of the ERα gene of SMCs. However,high concentrations (50rag/L) of ox-LDL induced demethylation,indicating that different risk factors of atherosclerosis with differentpotency might cause different aberrant methylation patterns in the promoter region of the ERα gene.Theatherogenic mechanism of Hcy might involve the hypermethylation of the ERα gene,leading to the proliferationof SMCs in atherosclerotic lesions.     

Changes in Photosystem Ⅱ Activity and Leaf Reflectance Features of Several Subtropical Woody Plants Under Simulated SO2 Treatment

The effects of simulated SO2 treatment on the photosynthetic apparatus were investigated in five subtropical forest plants, namely Plnus massonlana Lamb., Schlma superba Gardn. et Champ., Castanopsls flssa （Champ. ex Benth.） Rehd. et Wils., Acmena acuminatissima （BI.） Merr et Perry, and Cryptocarya concinna Hance. After leaf sections had been immersed in 0, 20, 50, and 100 mmol/L NaHSO3 for 20 h, total chlorophyll （Chl） content, Chl a/b, maximal photochemical efficiency, and the photochemical quantum yields of photosystem Ⅱ of all five woody plants were reduced to different degrees, whereas lutein content （Chl base） was increased. Two protective mechanisms, namely the xanthophyll cycle （de-epoxidation） and an anti-oxidant system （1,1-diphenyl-2-picrylhydrazyl radical-scavenging capacity）, showed differences in the degree of modulation under simulated SO2 treatment. Compared with control （distilled water treatment）, the revised normalized difference vegetation index, a leaf reflectance index, was lowered with Increasing concentrations of NaHSO3. Cryptocarya conclnna, a dominant species in the late succession stage of subtropical forests in South China, exhibited less sensitivity to NaHSO3. Conversely, Pinus massonlana, the pioneer hellophyte species, was most susceptible to NaHSO3 treatment. It Is suggested that SO2 pollution may accelerate the succession of subtropical forest.     

Effect of 2,4-D on Seedling Physiology and Cytogenetical Studies in Triticum aestivum and Phalaris minor (Gramineae)

Effects of 2, 4 -D on seedling growth and chromosomal abnormalities were studied in Triticum aestivum and Phalaris minor. Seeds were soaked at different concentrations of 2,4 -D （0.01%, 0.1%, 1.0% ） for 4, 8, 12 and 16 hours. 2,4-D suppressed the germination more severely in P. minor than in T. aestivum. Shoot and root length was retarded with the increase of concentration and time of treatment in both species. Generally radical was more negatively affected than coleoptile and emergence of radical was not observed at 1.0% concentration at 8, 12, and 16 hours of treatment in T. aestivum while in P. minor there was a total lack of radical emergence at 1.0% concentration for all durations of treatment. Stiff and curled roots and undifferentiated callus like scutellar tissues were observed in T. aestivum, while in P. minor the coleoptile obtained was lean, pale green in colour and was lying flat on filter paper. Mitotic index decreased, while chromosomal abnormalities, bridges and laggards were increased with the increase of concentration and soaking time however, laggards were not observed in T. aestivum. Clumping and chain formation of chromosomes at metaphase was also noticed in P. minor.     

Preparation and analysis of spermatocyte meiotic pachytene bivalents of pigs for gene mapping

Well-spread meiotic pachytene bivalents were obtained by using the prolonged hypotonic treatment com-bined with high chloroform Carnory’s fixative solution from cells of the testes of domestic pigs. Comparison in the division index and length of pachytene bivalents with metaphase chromosomes showed that those of the former are 5 times higher and 3.42(1.87-5.98) times longer than those of the latter. Comparative studies on chromomere maps of bivalents and mitotic chromosomal G-bands were conducted by using the chromo-some 12 as a example. Sex vesicle and various shapes of synaptic sex chromosomes have been observed.Two-color PRimed IN Situ (PRINS) labeling has been conducted successfully on pachytene bivalents of pigs.     

Response of transgenic potato seedlings to allelopathic pressure and the effect of nutrients in the culture medium

Chinese farmers frequently use a wheat–potato cropping system. The land area planted to transgenic potatoes is increasing because transgenic potatoes have greater resistance to pests and diseases. However, little is known about the bio-compatibility of transgenic potatoes with wheat straw. The objective of this tissue culture study was to determine the allelopathic effects of wheat straw on transgenic potato seedlings. Seedlings were cultured on normal MS medium (normal treatment) and nutrient-deficient MS medium (acclimated treatment) and then transferred to MS medium, which contained wheat straw powder. Wheat straw powder inhibited potato seedling growth in both treatments. Among the parameters analyzed in this study, inhibition was greatest for plant fresh weight and least for plant height. The inhibitive effects of wheat straw were greater for seedling roots compared to shoots. Resistance to allelopathic pressure from wheat straw was greater in acclimated seedlings compared to normal seedlings. This suggested that previous pressure may have induced tolerance in the transgenic potato seedlings. Furthermore, growth inhibition of potato seedlings from the normal treatment increased as the amount of wheat straw powder in the culture medium increased. Calculations indicated that the presence of wheat straw would lead to a 55% reduction in the total biomass of normal potato seedlings compared to a 39% reduction for acclimated seedlings. Parameters such as net photosynthesis rate (Pn) and quantum yield (Y(II)) changed as the nutrient content of the culture medium increased or decreased, but the changes in the parameters were smaller for acclimated seedlings compared to normal seedlings. This suggests that nutrient status during the culture period could help transgenic potato seedlings adapt and compensate for energy loss from seedlings in defending against allelopathic pressure. In summary, the results show that previous exposure to pressures such as nutrient deficiency may increase the allelopathic pressure resistance of transgenic potato seedlings.     

Response of transgenic potato seedlings to allelopathic pressure and the effect of nutrients in the culture medium

Chinese farmers frequently use a wheat–potato cropping system. The land area planted to transgenic potatoes is increasing because transgenic potatoes have greater resistance to pests and diseases. However, little is known about the bio-compatibility of transgenic potatoes with wheat straw. The objective of this tissue culture study was to determine the allelopathic effects of wheat straw on transgenic potato seedlings. Seedlings were cultured on normal MS medium (normal treatment) and nutrient-deficient MS medium (acclimated treatment) and then transferred to MS medium, which contained wheat straw powder. Wheat straw powder inhibited potato seedling growth in both treatments. Among the parameters analyzed in this study, inhibition was greatest for plant fresh weight and least for plant height. The inhibitive effects of wheat straw were greater for seedling roots compared to shoots. Resistance to allelopathic pressure from wheat straw was greater in acclimated seedlings compared to normal seedlings. This suggested that previous pressure may have induced tolerance in the transgenic potato seedlings. Furthermore, growth inhibition of potato seedlings from the normal treatment increased as the amount of wheat straw powder in the culture medium increased. Calculations indicated that the presence of wheat straw would lead to a 55% reduction in the total biomass of normal potato seedlings compared to a 39% reduction for acclimated seedlings. Parameters such as net photosynthesis rate (Pn) and quantum yield (Y(II)) changed as the nutrient content of the culture medium increased or decreased, but the changes in the parameters were smaller for acclimated seedlings compared to normal seedlings. This suggests that nutrient status during the culture period could help transgenic potato seedlings adapt and compensate for energy loss from seedlings in defending against allelopathic pressure. In summary, the results show that previous exposure to pressures such as nutrient deficiency may increase the allelopathic pressure resistance of transgenic potato seedlings.     

Root Morphology and Zn^2＋ Uptake Kinetics of the Zn Hyperaccumulator of Sedum alfredii Hance

Root morphology and Zn^2＋ uptake kinetics of the hyperaccumulating ecotype （HE） and nonhyperaccumulating ecotype （NHE） of Sedum alfredii Hance were investigated using hydroponic methods and the radiotracer flux technique. The results indicate that root length, root surface area, and root volume of NHE decreased significantly with increasing Zn^2＋ concentration in growth media, whereas the root growth of HE was not adversely affected, and was even promoted, by 500μmol/L Zn^2＋. The concentrations of Zn^2＋ in both ecotypes of S. alfredii were positively correlated with root length, root surface area and root volumes, but no such correlation was found for root diameter. The uptake kinetics for ^65Zn^2＋ in roots of both ecotypes of S. alfredii were characterized by a rapid linear phase during the first 6 h and a slower linear phase during the subsequent period of investigation. The concentration-dependent uptake kinetics of the two ecotypes of S. alfredii could be characterized by the Michaelis-Menten equation, with the Vmax for ^65Zn^2＋ influx being threefold greater in HE compared with NHE, indicating that enhanced absorption into the root was one of the mechanisms involved in Zn hyperaccumulation. A significantly larger Vmax value suggested that there was a higher density of Zn transporters per unit membrane area in HE roots.     

Joint effects of acetochlor and urea on germinating characteristics of crop seeds

In order to evaluate ecological risk of agrochemicals in common use, joint toxic effects of acetochlor and urea on germinating characteristics of Chinese cabbage (Brassica Pe-kinensis Rupr) seeds were investigated using the water-culture method and the soil-culture method. The results indicated(that excessive application of acetochlor and urea, when the concentrations were higher than 31.3 mg ·kg-1 for acetochlor and 500 mg ·kg-1 for urea, had strong inhibitory effects on the rate of seed germination, root elongation and hypocotyl length of Chinese cabbage. The inhibitory rate of the germinating characteristics of Chinese cabbage seeds was significantly increased with an increase in the concentration of acetochlor or urea. The two agrochemicals in water had a stronger toxicity than these in the soil at the same concentration. Among the three indexes, hypocotyl length was the most sensitive to the toxicity of acetochlor and urea.     

Concentrations of major elements and mercury in unstimulated human saliva

The aim of this study was a preliminary assessment of a possible role of human saliva in the diagnosis of some physiological and pathological changes in oral and body functions. Reliable procedures for collection and analysis of samples were established in order to assess total concentrations of Ca, K, Mg, Na, P, and Hg in whole unstimulated saliva. Possible relationships between element concentrations and sex, age, smoking, illness conditions, or side effects resulting from the use of drugs were investigated. The effects of stimulated or unstimulated collection procedures, dental prosthesis, and amalgam fillings were also evaluated. Total concentrations of major cations and Hg in whole saliva from 33 healthy adults living in the Siena district showed a coefficient of variation ranging from 11% (P) to 53% (Na) and average values were in the same range of those previously reported for unstimulated saliva. Healthy males had significantly higher concentrations of K, Na, P, and Na/K, Na/Ca, Na/Mg, and Na/P values than females. Age, smoking, dental prosthesis, and amalgam fillings had no significant effects on the concentrations of major elements. On the contrary, concentrations of Hg were positively correlated to the number of amalgam fillings and increased at a rate of about 1.9 μg/L for each filling. No correlations were found between Hg concentrations and those of major elements. Comparisons with literature data showed a different composition (particularly for Na and Hg concentrations) between unstimulated and stimulated saliva. Samples from patients affected by idiopathic pulmonary fibrosis had significantly higher concentrations of K and the maximum value was measured in a patient affected by acute pulmonary edema. This increase was likely the result of pharmacological treatments with tricyclic antidepressants and/or saline solutions. Data reported in this study, although preliminary, contribute to the assessment of levels of major cations (some of them very little investigated) and Hg in whole unstimulated human saliva and provides consistent support for further research on the possible use of this easy accessible matrix as a diagnostic tool of body function changes.     

Effects of the Application of Different Concentrations of NaN3 for Different Times on the Morphological and Cytogenetic Characteristics of Barley (Hordeum vulgare L.) Seedlings

Sodium azide (NaN3) has been used in many biological studies as a mutagen. In the present study, the morphological and cytogenetic effects of NaN3 on barley (Hordeum vulgare L.) seedlings were investigated. Seeds of barley were treated with different concentrations of NaN3 (0.5, 1.0, 1.5, 2.0, 2.5, and3.0 mmol/L) and applied for different periods of time (3 and 4 h). Parameters investigated, except for the mitotic index, were determined on Days 7 and 14. Increasing concentrations of NaN3 affected germination rates on Days 7 and 14 following application for 3 and 4 h. Although the length of the roots and leaves was affected by treatment with NaN3 on Day 14 of the germination period, coleoptile length was affected by NaN3 treatment on Day 7. Increasing concentrations of NaN3 and increased treatment period decreased the mitotic index compared with the untreated control. The inhibitory effects of NaN3 on the mitotic index indicate that NaN3 can have genotoxic and mutagenic effects on barley seedlings.     

Warming and elevated CO2 affect the relationship between seed mass, germinability and seedling growth in Austrodanthonia caespitosa, a dominant Australian grass

While the influence of elevated CO₂ on the production, mass and quality of plant seeds has been well studied, the effect of warming on these characters is largely unknown; and there is practically no information on possible interactions between warming and elevated CO₂, despite the importance of these characters in population maintenance and recovery. Here, we present the impacts of elevated CO₂ and warming, both in isolation and combination, on seed production, mass, quality, germination success and subsequent seedling growth of Austrodanthonia caespitosa , a dominant temperate C₃ grass from Australia, using seeds collected from the TasFACE experiment. Mean seed production and mass were not significantly affected by either elevated CO₂ or warming, but elevated CO₂ more than doubled the proportion of very light, inviable seeds ( P < 0.05) and halved mean seed N concentration ( P < 0.04) and N content ( P < 0.03). The dependence of seed germination success on seed mass was affected by an elevated CO₂× warming interaction ( P < 0.004), such that maternal exposure to elevated CO₂ or warming reduced germination if applied in isolation, but not when applied in combination. Maternal effects were retained when seedlings were grown in a common environment for 6 weeks, with seedlings descended from warmed plants 20% smaller ( P < 0.008) with a higher root : shoot ratio ( P < 0.001) than those from unwarmed plants. Given that both elevated CO₂ and warming reduced seed mass, quality, germinability or seedling growth, it is likely that global change will reduce population growth or distribution of this dominant species.     

The effect of nitrogen source on photosynthesis of carob at high CO2 concentrations

Carob seedlings ( Ceratonia siliqua L. cv. Mulata), fed with nitrate or ammonium, were grown in growth chambers containing two levels of CO₂ (360 or 800 μl l⁻¹), three root temperatures (15, 20 or 25°C), and the same shoot temperature (20/24°C, night/day temperature). The response of the plants to CO₂ enrichment was affected by environmental factors such as the type of inorganic nitrogen in the medium and root temperature. Increasing root temperature enhanced photosynthesis rate more in the presence of nitrate than in the presence of ammonium. Differences in photosynthetic products were also observed between nitrate- and ammonium-fed carob seedlings. Nitrate-grown plants showed an enhanced content of sucrose, while ammonium led to enhanced storage of starch. Increase in root temperature caused an increase in dry mass of the plants of similar proportions in both nitrogen sources. The enhancement of the rates of photosynthesis by CO₂ enrichment was proportionally much larger than the resulting increases in dry mass production when nitrate was the nitrogen source. Ammonium was the preferred nitrogen source for carob at both ambient and high CO₂ concentrations. The level of photosynthesis of a plant is limited not only by atmospheric CO₂ concentration but also by the nutritional and environmental conditions of the root.     

Interaction between Pfr and growth substances in the germination of light-requiring Kalanchoë seeds

Seeds of Kalanchoë blossfeldiana Poelln. cv. Feucrblüte, incubated on gibberellic acid, become very light-sensitive through a synergism between the far-red absorbing form of phytochrome and the growth substance, which results in high physiological activity of short far-red (FR) exposures. On 2 × 10^-3 M gibberellic acid (GA₃), one saturating FR pulse is somewhat more effective than one saturating red light (R) irradiation. Fluence-response curves for R and FR confirm this observation. At lower GA₃ concentrations, this difference disappears and the effects of one saturating R and FR pulse decrease in an identical way with the GA₃ concentration. When two saturating irradiations, separated by 24 h are given, the effect of FR falls off faster than that of R at low GA₃ concentrations. Consequently, the second irradiation must have a different impact in comparison with the first one. Of the other growth substances tested, only a mixture of gibberellins A4 and A₇ had an analogous, still more pronounced effect than GA₃. Abscisic acid (ABA) inhibits the phytochrome-mediated germination of Kalanchoë , both in the absence and presence of GA₃. An antagonism between ABA and GA₃ was demonstrated.     

Effects of elevated ozone on photosynthesis and stomatal conductance of two soybean varieties: a case study to assess impacts of one component of predicted global climate change

Global climatic change scenarios predict a significant increase in future tropospheric ozone (O₃) concentrations. The present investigation was done to assess the effects of elevated O₃ (70 and 100 ppb) on electron transport, carbon fixation, stomatal conductance and pigment concentrations in two tropical soybean ( Glycine max L.) varieties, PK 472 and Bragg. Plants were exposed to O₃ for 4 h·day⁻¹ from 10:00 to 14:00 from germination to maturity. Photosynthesis of both varieties were adversely affected, but the reduction was higher in PK 472 than Bragg. A comparison of chlorophyll a fluorescence kinetics with carbon fixation suggested greater sensitivity of dark reactions than light reactions of photosynthesis to O₃ stress. The O₃-induced uncoupling between photosynthesis and stomatal conductance in PK 472 suggests the reduction in photosynthesis may be attributed to a factor other than reduced stomatal conductance. An increase in internal CO₂ concentration in both O₃-treated soybean varieties compared suggests that the reduction in photosynthesis was due to damage to the photosynthetic apparatus, leading to accumulation of internal CO₂ and stomatal closure. The adverse impact of O₃ stress increased at higher O₃ concentrations in both soybean varieties leading to large reductions in photosynthesis. This study suggests that O₃-induced reductions in photosynthesis in tropical and temperate varieties are similar.     

Cadmium and copper change root growth and morphology of Pinus pinea and Pinus pinaster seedlings

Heavy metal loads in forest soils have been increasing over time due to atmospheric inputs. Accumulation in the upper soil layers could affect establishment of seedlings and forest regeneration. Mediterranean species show a high initial root development, allowing seedlings to reach the moisture of deeper soil layers. In the present work seedlings of stone pine ( Pinus pinea L.) and maritime pine ( Pinus pinaster Ait.), were grown in culture solution supplied with 0.0, 0.1, 1 or 5 μ M CdSO₄ or with 1 μ M CdSO₄ and 1 μ M CuSO₄ combined. In both species tap-root elongation was drastically reduced in the 5 μ M Cd²⁺ and in the (Cd²⁺+ Cu²⁺) treatments. A supply of 0.1 or 1 μ M Cd²⁺, however, enhanced root elongation in Pinus pinea without significantly influencing root elongation in Pinus pinaster . In both species the root density (weight per unit length) and the width of the cortex increased in response to Cd²⁺ exposure. In Pinus pinaster the mitotic index decreased at the higher Cd²⁺ concentrations and when Cd²⁺ and Cu²⁺ were combined. The data suggest that cell elongation is more sensitive to Cd²⁺ than cell division. The number and length of the lateral roots were also affected by Cd²⁺ treatment to a higher degree in Pinus pinaster than in Pinus pinea, reflecting the different Cd- tolerance of the two species.     

Photoinhibition of respiratory CO2 release in the green alga Scenedesmus

¹⁴CO₂ evolution of prelabeled Scenedesmus obliquus Kütz, has been followed in the dark and in the light. In the light, no carbon dioxide is evolved. Addition of unlabeled NaHCO, leads to ¹⁴CO₂ release attaining 20 to 30% of the dark rate. Double-reciprocal plots of NaHCO₃ concentrations vs ¹⁴CO₂ release results in a straight line, indicative of competition between exogenously supplied bicarbonate and endogenously evolved carbon dioxide. With this method, it is possible to measure CO₂ evolved by respiration in the light and to show that true photoinhibition of respiration occurs in Scenedesmus . In the light. DCMU substantially increases ¹⁴CO₂ evolution; in the presence of the uncoupler carbonyl cyanide- m -chlorophenylhydrazone. ¹⁴CO₂ evolution is comparable to that in the dark. ¹⁴CO₂ release and oxygen uptake in the dark are only slightly affected by cyanide, indicative of a cyanide-resistant respiration and/or fermentation as the essential CO₂-yielding processes in the presence of cyanide. These results, compared with concurrent ATP levels, lead us to assume that energy charge is not the only factor responsible for photoinhibition of respiration.     

Monocytic differentiation of HL60 cells induced by potent analogs of vitamin D3 precedes the G1/G0 phase cell cycle block

Abstract. Differentiation of mammalian cells is accompanied by reduced rates of proliferation and an exit from the cell cycle. Human leukemic cells HL60 present a widely used model of neoplastic cell differentiation, and acquire the monocytic phenotype when exposed to analogs of vitamin D₃ (VD₃). The maturation process is accompanied by two blocks in the cell cycle: an arrest in the G₁/G₀ phase, and a recently described G₂+ M block. In this study we have analyzed the traverse of the cell cycle phases of the well-differentiating HL60-G cells exposed to one of ten analogs of VD₃, and compared the cell cycle effects of each compound with its potency as a differentiation-inducing agent. We found that in general there was a good correlation between the effects of these compounds on the cell cycle and on differentiation, but the best cell cycle predictor of differentiation potency was the extent of accumulation of the cells in the G₂ compartment. All analogs induced a marked decrease in the mitotic index, and polynucleation of HL60 cells was produced, especially by compounds which were effective as inducers of differentiation. Time course studies showed that induction of differentiation was accompanied by a transient increase of the proportion of cells in the G₂+ M compartment, but preceded the G₁ to S, and the G₂ compartment blocks. These studies indicate that complex changes in the cell cycle traverse accompany, but do not precede, the acquisition of the monocytic phenotype by HL60 cells.     

Induction of nitrate reductase in needles of Scots pine seedlings by NOX and NO3−

The possibility to induce nitrate reductase (NR; EC 1.6.6.2) in needles of Scots pine ( Pinus sylvestris L.) seedlings was studied. The NR activity was measured by an in vivo assay. Although increased NR activities were found in the roots after application of NO₃⁻, no such increase could be detected in the needles. Detached seedlings placed in NO₃⁻ solution showed increasing NR activities with increasing NO₃⁻ concentrations. Exposure of seedlings to NO_x (70–80 ppb NO₂ and 8–12ppb NO) resulted in an increase of the NR activity from 10–20 nmol NO₂⁻ (g fresh weight)⁻¹ h⁻¹ to about 400 nmol NO₂⁻ (g fresh weight)⁻¹ h⁻¹. This level was reached after 2–4 days of exposure, thereafter the NR activity decreased to about 200 nmol NO₂⁻ (g fresh weight)⁻¹ h⁻¹. Analyses of free amino acids showed low concentrations of arginine and glutamine in NO_x-fumigated seedlings compared to corresponding controls.     

ATPase activity associated with vacuoles and tonoplast vesicles isolated from the CAM plant, Kalanchoë daigremontiana

Intact vacuoles were isolated from leaves of the CAM plant, Kalanchoë daigremontiana Hamet et Perr. Both ATPase and acid phosphatase activities were found in the vacuoles. Purified tonoplast vesicles showed only ATPase activity with a pH optimum of 8.0. This activity was Mg²⁺-dependent and KCI or NaCI caused a further stimulation. N,N'-dicyclohexylcarbodiimide, diethylstilbestrol and quercetin inhibited the ATPase almost completely at concentrations well below 1 m M. NaVo₃, 1-ethyl-3(3-dimethylaminopropyl)carbodiimide, oligomycin and NaN₃ had little or no effect. Carbonyl cyanide m -chlorophenylhydrazone stimulated the ATPase about 40% at 5 × 10⁻⁴ M. The K_m for ATP was found to be 0.55 m M. These results indicate that the ATPase found in the tonoplast membrane of Kalanchoë daigremontiana is qualitatively similar to that of other plant species.