Changes in Stomatal Conductance in Intact Ageing Wheat Leaves in Response to Abscisic Acid

The characteristics of ABA-induced changes in the fluxes ofCO₂ and water vapour from whole leaves of spring wheat (Triticumaestivum cv. Wembley) were examined. Aqueous solutions of ABAwere supplied via the transpiration stream to intact leavesof different ages mounted within a gas exchange cuvette. ABA caused a reduction in stomatal conductance (g) that wasproportional to the concentration in the solution fed to theleaf. For the maintenance of a reduction in g there was a requirementfor a continual supply of ABA. At concentrations greater than10^–2 mol m^–3 ABA reduced g by at least 50% of thecontrol value, while 1.0 mol m^–3 closed stomata within2 h. Concentrations as low as 10^–3 mol m^–3 produceda 20% reduction in g. As leaves aged they became less responsiveto applied ABA. The possibility that the stomatal response may change aftera leaf has previously experienced a pulse of ABA was exploredby repeating the exposure of a leaf to 10^–2 mol m^–3ABA. The first pulse of ABA produced a greater reduction ing than a subsequent exposure the following day. This declinein response of g to ABA on repeated exposure was maintainedwith leaves of different ages. The characteristics of the stomatal response to ABA are discussedin the context of what is known about the location of receptorsfor the hormone. It seems likely that a failure to respond toABA that has previously accumulated in the guard cells shouldbe viewed by means of maximizing the sensitivity to the currentsupply of ABA. It is suggested that the smaller response ofthe stomata of older leaves to ABA makes them more susceptibleto water stress, so that they can act as sensors for decliningwater potentials to give early protection to younger, metabolicallyactive leaves. Key words: Abscisic acid, leaf age, stomatal conductance, Triticum aestivum     

完整叶片内玉米黄质的分光光度分析

以载玻片作参比材料,成功地解决了较厚叶片不能直接测定505nm光吸收来反映玉米黄质水平的难题。并通过对银杏叶片505nm光吸收的不同方法测定分析比较,表明该方法测定结果稳定可靠。     

完整叶片内玉米黄质的分光光度分析

以载玻片作参比材料,成功地解决了较厚叶片不能直接测定505 nm光吸收来反映玉米 黄质水平的难题。并通过对银杏叶片505 nm光吸收的不同方法测定分析比较,表明该方法测 定结果稳定可靠。     

The Effect of Benzyladenine on Changes in Nuclease and Protease Activities in Intact Bean Leaves during Ageing

Primary leaves of intact bean plants (Phaseolus vulgaris L.) were treated with benzyladenine (BA) at different stages during growth. Changes in DNase, RNase, and proteas activities in the leaves were followed. Unlike the case of various excised tissues, cytokinin raised the activities of these hydrolases in intact bean leaves. Because BA elevated the levels of DNA, RNA, and protein in intact leaves, it may stimulate both synthesis and decomposition of these cellular constituents. The hydrolase activities showed differential responses to BA according to the age at which the leaf received the hormone treatment.     

Thermal Tolerance of Zymomonas mobilis: Temperature-Induced Changes in Membrane Composition

The membrane composition of Zymomonas mobilis changed dramatically in response to growth temperature. With increasing temperature, the proportion of vaccenic acid declined with an increase in myristic acid, the proportion of phosphatidylcholine and cardiolipin increased with decreases in phosphatidylethanolamine and phosphatidylglycerol, and the phospholipid/protein ratio of the membrane declined. These changes in membrane composition were correlated with changes in thermal tolerance and with changes in membrane fluidity. Cells grown at 20°C were more sensitive to inactivation at 45°C than were cells grown at 30°C, as expected. However, cells grown at 41°C (near the maximal growth temperature for Z. mobilis) were hypersensitive to thermal inactivation, suggesting that cells may be damaged during growth at this temperature. When cells were held at 45°C, soluble proteins from cells grown at 41°C were rapidly lost into the surrounding buffer in contrast to cells grown at lower temperatures. The synthesis of phospholipid-deficient membranes during growth at 41°C was proposed as being responsible for this increased thermal sensitivity.     

Temperature-Induced Changes in the Sporicidal Activity and Chemical Properties of Glutaraldehyde

Freshly prepared 2% acid and alkaline glutaraldehyde solutions were stored at 4, 20, and 37 C. At intervals, samples were removed and changes in pH, ultraviolet spectrum, and sporicidal activity (against Bacillus pumilus spores) were recorded. Alkaline solutions stored at 4 C showed little changes in these properties, whereas such solutions stored at 37 C became turbid and showed a decrease in pH, marked changes in ultraviolet spectrum, and an almost complete loss of sporicidal activity. Intermediate results were obtained with alkaline solutions stored at 20 C. In contrast, acid 2% glutaraldehyde solutions (initial pH 3.5) showed comparatively few changes in their properties. Treatment of spores with freshly prepared glutaraldehyde solutions (0.5%) at temperature above 40 C reduced the effect of pH on sporicidal activity.     

Effect of Benzyladenine on Diurnal Changes in DNA Content per Chloroplast and Chloroplast Replication in Intact Bean Leaves

The effect of benzyladenine (BA) on the diurnal changes in DNAand Chl contents per chloroplast and chloroplast replicationin primary leaves of bean plants (Phaseolus vulgaris L.) grownunder a 16 h light/8 h dark cycle was studied. Experiments weremade on primary leaves in the early expansion phase, where celldivision had been completed but chloroplasts were replicating.In untreated controls, chloroplast number, Chl content and freshweight per leaf showed daily periodic changes. Chl content perchloroplast increased in the light period every day, and freshweight per leaf increased most rapidly in the early dark period.Chloroplast number per leaf increased rapidly in the early darkperiod on day 9, though the increase began a little earlierand was less sharp on days 8 and 10. During these periods, DNAcontent per chloroplast was decreasing due to chloroplast divisionas chloroplast DNA (ctDNA) per leaf remained unchanged throughoutthe experimental period. BA induced increases in Chi contentper chloroplast, ctDNA content and fresh weight per leaf within6 h of its application, regardless of whether it was appliedat or 10 h after the beginning of the light period. Applicationof BA at 10 h in the light period shifted the start of chloroplastreplication by 6 h compared to that in untreated controls. However,when BA was applied at the beginning of illumination, the startof chloroplast replication showed the same relative change intime as above. 5-Fluorodeoxyuridine (5-FdU) promptly preventedBA-induced increase in Chl content and chloroplast number perleaf as well as ctDNA content per leaf.     

Regulation of Purine Metabolism in Intact Leaves of Coffea arabica

The capacity of Coffea arabica leaves (5- x 5-mm pieces) to synthesize de novo and catabolize purine nucleotides to provide precursors for caffeine (1,3,7-trimethylxanthine) was investigated. Consistent with de novo synthesis, glycine, bicarbonate, and formate were incorporated into the purine ring of inosine 5[prime]-monophosphate (IMP) and adenine nucleotides ([sigma]Ade); azaserine, a known inhibitor of purine de novo synthesis, inhibited incorporation. Activity of the de novo pathway in C. arabica per g fresh weight of leaf tissue during a 3-h incubation period was 8 [plus or minus] 4 nmol of formate incorporated into IMP, 61 [plus or minus] 7 nmol into [sigma]Ade, and 150 nmol into caffeine (the latter during a 7-h incubation). Coffee leaves exhibited classical purine catabolism. Radiolabeled formate, inosine, adenosine, and adenine were incorporated into hypoxanthine and xanthine, which were catabolized to allantoin and urea. Urease activity was demonstrated. Per g fresh weight, coffee leaf squares incorporated 90 [plus or minus] 22 nmol of xanthine into caffeine in 7 h but degraded 102 [plus or minus] 1 nmol of xanthine to allantoin in 3 h. Feedback control of de novo purine biosynthesis was contrasted in C. arabica and Cucurbita pepo, a species that does not synthesize purine alkaloids. End-product inhibition was demonstrated to occur in both species but at different enzyme reactions.     

Temperature-Induced Changes in Lipid Fluidity Alter the Conformation of Proteins in Senescing Plant Membranes

Complementary biophysical techniques have provided evidencefor an effect of temperature-induced changes in lipid fluidityon the conformation of proteins in senescing plant membranes.Smooth microsomal membranes were isolated from bean cotyledons(Phaseolus vulgaris L. (cv Kinghorn)) at various stages of senescence.Lipid fluidity was measured by fluorescence polarization afterlabelling the membranes with diphenyl hexatriene (DPH). Alterationsin protein conformation were determined by labelling the membraneswith the paramagnetic sulfhydryl reagent, 3-maleimido proxyl(3-MP), and following changes in the ratio of weakly immobilizedto strongly immobilized (w/s) spin label. Plots of w/s as afunction of temperature featured characteristic break pointsfor each age of membrane. Corresponding break points at virtuallyidentical temperatures were also observed in plots of DPH polarizationversus temperature for membranes as well as for liposomes preparedfrom lipid extracts of membranes. In at least one instance thesebreak points in DPH polarization did not correspond to liquid-crystallineto gel phase transitions in the lipid. The fluidity of microsomalmembranes decreased with advancing senescence of the cotyledons,and there were also changes in the parameter w/s for membraneproteins with advancing age. The results indicate that subtlechanges in the molecular ordering of lipid bilayers alter therelative proportions of weakly and strongly immobilized sulfhydrylgroups in the membrane proteins, which can be interpreted asreflecting changes in protein conformation. ¹ Present address: Agronomy Department, Cornell University,Ithaca, N.Y. 14853, U.S.A. (Received January 19, 1987; Accepted April 14, 1987)     

Intact Leaves Exhibit a Climacteric-Like Rise in Ethylene Production before Abscission

The rate of ethylene production by intact, attached leaves of cotton plants (Gossypium hirsutum L.) during aging and senescence was studied using a continuous flow system that allowed air around enclosed leaves to be scrubbed to collect and assay ethylene. Senescence of lower leaves began around 150 d after planting in a controlled environment room. A progressive decline in the ethylene production rate was observed when comparing the 3rd, 6th, and 10th leaves from the base with each other. Ethylene production rates of individual leaves also declined over a 50-d period. However, as leaves began to appear chlorotic, a peak of ethylene production occurred that lasted for about 4 d followed by abscission. This peak involved a 3-fold or greater increase in the rate of ethylene production. The data indicate that intact leaves experience a climacteric-like surge in ethylene production after visible symptoms of senescence appear. This “ethylene climacteric” is apparently the signal that initiates hydrolysis of cell walls in the abscission zone.     

Limitations of Gas Exchanges in Intact Leaves during Ontogeny of Field-grown Chickpeas

The ontogenic changes in several component processes of photosynthesiswere measured in chickpeas. Gas exchange characteristics ofintact leaves were studied to analyse the effects of ambientconditions under which chickpeas are usually grown. The CO₂assimilation rate per unit leaf area remained fairly high duringthe vegetative stage, reaching a peak at early pod-fill anddeclining subsequently throughout pod development. The intercellularCO₂ partial pressure (C₁) remained more or less constant (195µbar) during vegetative growth and the early stages ofseed-filling. With falling RWC and PAR interception, the stomatalconductance declined more rapidly than the CO₂ assimilationrate resulting in a value of C₁ less than that normally existingunder ambient conditions. From the A/C₁-analysis, CO₂ assimilationduring pod-filling appears to be limited by the RuBP-regenerationcapacity because the carboxylation efficiency and in vitro RuBPCaseactivity were initially unaffected. However, as leaves aged,the carboxylation efficiency and in vitro RuBPCase activitydecreased abruptly with increasing leaf temperatures above 30°C, and the C₁ was greater than normally existing values(195 µbar), suggesting an increased mesophyll limitationof photosynthesis. It is suggested that a decline in the CO₂assimilation rate of leaves during pod development and an acceleratedsenescence are induced by adverse ambient conditions, particularlyplant water stress and high leaf temperature. Key words: Cicer arietinum L., gas exchange, photosynthesis, ribulose-1,5-bisphosphate carboxylase     

Amplification of Extrachromosomal Circular DNA in Intact Bean Leaves Treated with Benzyladenine

Bean (Phaseolus vulgaris L.) leaves were treated with benzyladenine(BA) beginning 7 days after sowing, and were harvested at 21days. There were less than 10 molecules per cell of small polydispersecircular (spc) DNA in mesophyll cells from untreated leaves,and more than 30 molecules in the BA-treated cells. The spcDNAranged in size from 0.2 to 7.5 µm, with a mean lengthof 1.7 µm. (Received May 29, 1985; Accepted December 24, 1985)     

Temperature-Induced Changes in the Fatty Acid Composition of the Cyanobacterium, Synechocystis PCC6803

Changes in glycerolipid and fatty acid composition with a change in growth temperature were studied in the cyanobacterium, Synechocystis PCC6803. Under isothermal growth conditions, temperature did not significantly affect the composition of the various classes of lipids, but a decrease in temperature altered the degree of unsaturation of C₁₈ acids at the sn-1 position, but not that of C₁₆ acids at the sn-2 position of the glycerol moiety in each class of lipids. When the growth temperature was shifted from 38°C to 22°C, the desaturation of C₁₈ acids, but not that of C₁₆ acids, was stimulated. The desaturation of fatty acids occurred only in the light and was inhibited by chloramphenicol, rifampicin and 3-(3,4-dichlorophenyl)-1, 1-dimethylurea, but not by cerulenin, an inhibitor for fatty acid synthesis. These findings suggest that desaturase activities are induced after a shift from a higher to a lower temperature, and that the desaturation of fatty acids is connected with the reactions involved in photosynthetic electron transport.     

Photosynthesis and Ribulose 1,5-Bisphosphate Concentrations in Intact Leaves of Xanthium strumarium L

The interacting effects of the rate of ribulose 1,5-bisphosphate (RuBP) regeneration and the rate of RuBP utilization as influenced by the amount and activation of RuBP carboxylase on photosynthesis and RuBP concentrations were resolved in experiments which examined the kinetics of the response of photosynthesis and RuBP concentrations after step changes from a rate-saturating to a rate-limiting light intensity in Xanthium strumarium. Because RuBP carboxylase requires several minutes to deactivate in vivo, it was possible to observe the effect of reducing the rate of RuBP regeneration on the RuBP concentration at constant enzyme activation state by sampling very soon after reducing the light intensity. Samples taken over longer time periods showed the effect of changes in enzyme activation at constant RuBP regeneration rate on RuBP concentration and photosynthetic rate. Within 15 s of lowering the light intensity from 1500 to 600 microEinsteins per square meter per second the RuBP concentration in the leaves dropped below the enzyme active site concentration, indicating that RuBP regeneration rate was limiting for photosynthesis. After longer intervals of time, the RuBP concentration in the leaf increased as the RuBP carboxylase assumed a new steady state activation level. No change in the rate of photosynthesis was observed during the interval that RuBP concentration increased. It is concluded that the rate of photosynthesis at the lower light intensity was limited by the rate of RuBP regeneration and that parallel changes in the activation of RuBP carboxylase occurred such that concentrations of RuBP at steady state were not altered by changes in light intensity.     

拟南芥连体和离体叶片光合作用的光响应

测定出 2 5 0 μmol·m-2 ·s-1光强下培养的整株拟南芥连体叶片的光合作用在光强为 80 0 μmol·m-2 ·s-1左右达到光饱和 ,其离体叶片光合作用对光强响应的结果与此类似。自然条件下生长的珊瑚树连体与离体叶片光合作用均在光强约为 10 0 0 μmol·m-2 ·s-1下达到饱和 ,这验证所测得的拟南芥连体和离体叶片光合作用光响应的结果是正确的     

Low Night Temperature-Induced Changes in Photosynthesis and Rubber Accumulation in Guayule (Parthenium Argentatum Gray)

Photosynthetica - Three-year-old plants of Parthenium argentatum Gray cv. 11591 grown under natural photoperiod were exposed for 60 d to low night temperature (LNT) of 15 °C (daily from 18:00...     

Luminescence Induced in Intact Leaves and Isolated Chloroplasts by Alcohols and Other Substances

Elodea leaves and spinach chloroplasts emit red light when treated with alcohols or certain other solvents for chlorophyll. The intensity of the light, the lag phase and the threshold concentration vary considerably between different alcohols. Light emission from a leaf starts a few seconds to a few minutes after the addition of alcohol to the medium, reaches a maximum after 2–45 minutes (sometimes more) and then continues for many hours. Despite the faintness of the glow, the total number of photons given off from a leaf after addition of alcohol may exceed the number of photons given off from the same sample as long-lived afterglow after saturating irradiation with far-red light. The maximum yield of photons per chlorophyll molecule is a little more than 10^–5. The alcohol-induced luminescence is not influenced by a decrease in the oxygen tension to one fifth of the normal. Electron micrographs of treated leaves reveal that the thylakoid lipids contract to drops at the edges of the grana. Treatment of isolated chloroplasts with ethanol results in light emission when the concentration is high enough to dissolve the chlorophyll. It is estimated that the surface free energy in the thylakoid lipid-aqueous interface, due to ordinary interfacial tension, is large enough to account for the light emission observed when the interfaces contract or disappear.     

Method to Obtain a Chlorophyll-free Preparation of Intact Mitochondria from Spinach Leaves

Mitochondria from green leaves of spinach have been prepared using a three-step procedure involving differential centrifugation, partition in an aqueous dextran polyethylene glycol two-phase system and Percoll gradient centrifugation. The mitochondrial fractions after the different steps of purification were compared. The final mitochondrial preparation was totally free from chloroplast material measured as chlorophyll content. The enrichment of mitochondria in relation to peroxisomes and microsomes was approximately 12 and 33 times, respectively, based on NAD:isocitrate dehydrogenase activity, glycolate oxidase activity, and NADPH:cytochrome c oxidoreductase activity. The apparent intactness of the inner and the outer mitochondrial membranes was higher than 90% as measured by latency of enzyme activities. The mitochondria showed high respiratory rates with respiratory control and the ADP/O ratios approached the theoretical limits.