Photosynthate Partitioning into Starch in Soybean Leaves: I. Effects of Photoperiod versus Photosynthetic Period Duration

Photosynthesis, photosynthate partitioning into foliar starch, and translocation were investigated in soybean plants (Glycine max (L.) Merr. cv. Amsoy 71), grown under different photoperiods and photosynthetic periods to determine the controls of leaf starch accumulation. Starch accumulation rates in soybean leaves were inversely related to the length of the daily photosynthetic period under which the plants were grown. Photosynthetic period and not photoperiod per se appears to be the important factor. Plants grown in a 14-hour photosynthetic period partitioned approximately 60% of the daily foliar accumulation into starch whereas 7-hour plants partitioned about 90% of their daily foliar accumulation into starch. The difference in starch accumulation resulted from a change in photosynthate partitioning between starch and leaf residual dry weight. Residual dry weight is defined as leaf dry weight minus the weight of total nonstructural carbohydrates. Differences in photosynthate partitioning into starch were also associated with changes in photosynthetic and translocation rates, as well as with leaf and whole plant morphology. It is concluded that leaf starch accumulation is a programmed process and not simply the result of a limitation in translocation.     

Control of the Acclimation of Photosynthesis to Light and Temperature in Relation to Partitioning of Photosynthate in Developing Soybean Leaves

Photosynthetic acclimation was examined by exposing third trifoliolateleaves of soybeans to air temperatures of 20 to 30°C andphotosynthetic photon flux densities (PPFD) of 150 to 950µmolphotons m^–2 s^–1 for the last 3 d before they reachedmaximum area. In some cases the environment of the third leafwas controlled separately from that of the rest of the plant.Photosynthesis, respiration and dry mass accumulation were determinedunder the treatment conditions, and photosynthetic capacity,and dry mass and protein content were determined at full expansion.Photosynthetic capacity, the light-saturated rate of net carbondioxide exchange at 25°C and 34 Pa external partial pressureof carbon dioxide, could be modified between 21 and 35 µmolCO₂ m^–2 s^–1 by environmental changes after leaveshad become exporters of photosynthate. Protein per unit leafmass did not differ between treatments, and photosynthetic capacityincreased with leaf mass per unit area. Photosynthetic capacityof third leaves was affected by the PPFD incident on those leaves,but not by the PPFD on other leaves on the plant. Photosyntheticcapacity of third leaves was affected by the temperature ofthe rest of the plant, but not by the temperature of the thirdleaves. Photosynthetic capacity was linearly related to carbondioxide exchange rate in the growth regimes, but not to daytimePPFD. At high PPFD, and at 25 and 30°C, mass accumulationwas about 28% of the mass of photosynthate produced. At lowerPPFD, and at 20°C, larger percentages of the photosynthateproduced accumulated as dry mass. The results suggest that photosynthatesupply is an important factor controlling leaf structural growthand, consequently, photosynthetic acclimation to light and temperature. Key words: Glycine max (L.) Merr., photosynthesis, temperature acclimation, light acclimation, photosynthate partitioning     

Biochemical Basis for Partitioning of Photosynthetically Fixed Carbon between Starch and Sucrose in Soybean (Glycine max Merr.) Leaves

The control of photosynthetic starch/sucrose formation in leaves of soybean (Glycine max L. Merr.) cultivars was studied in relation to stage of plant development, photosynthetic photoperiod, and nitrogen source. At each sampling, leaf tissue was analyzed for starch content, activities of sucrose-metabolizing enzymes, and labeling of starch and sucrose (by ¹⁴CO₂ assimilation) in isolated cells. In three of the four varieties tested, nodulated plants had lower leaf starch levels and higher activities of sucrose phosphate synthetase (SPS), and isolated mesophyll cells incorporated more carbon (percentage of total ¹⁴CO₂ fixed) into sucrose and less into starch as compared to nonnodulated (nitrate-dependent) plants. The variation among cultivars and nitrogen treatments observed in the activity of SPS in leaf extracts was positively correlated with labeling of sucrose in isolated cells (r = 0.81) and negatively correlated with whole leaf starch content (r = −0.66). The results suggested that increased demand for assimilates by nodulated roots may be accommodated by greater partitioning of carbon into sucrose in the mesophyll cells. We have also confirmed the earlier report (Chatterton, Silvius 1979 Plant Physiol 64: 749-753) that photoperiod affects partitioning of fixed carbon into starch. Within two days of transfer of nodulated soybean Ransom plants from a 14-hour to a 7-hour photoperiod, leaf starch accumulation rates doubled, and this effect was associated with increased labeling of starch and decreased labeling of sucrose in isolated cells. Concurrently, activities of SPS, sucrose synthase, and uridine diphosphatase in leaves were decreased.     

Euploidy in Ricinus: EUPLOIDY EFFECTS ON PHOTOSYNTHETIC ACTIVITY AND CONTENT OF CHLOROPHYLL-PROTEINS

The effects of nuclear genome duplication on the chlorophyll-protein content and photochemical activity of chloroplasts, and photosynthetic rates in leaf tissue, have been evaluated in haploid, diploid, and tetraploid individuals of the castor bean, Ricinus communis L. Analysis of this euploid series revealed that both photosystem II (2,6-dichlorophenolindophenol reduction) and photosystem I oxygen uptake (N,N,N′,N′-tetramethyl-p-phenylenediamine to methyl viologen) decrease in plastids isolated from cells with increasingly larger nuclear complement sizes. Photosynthetic O₂-evolution and ¹⁴CO₂-fixation rates in leaf tissue from haploid, diploid, and tetraploid individuals were also found to decrease with the increase in size of the nuclear genome. Six chlorophyll-protein complexes, in addition to a zone of detergent complexed free pigment, were resolved from sodium dodecyl sulfate-solubilized thylakoid membranes from cells of all three ploidy levels. In addition to the P700-chlorophyll a-protein complex and the light-harvesting chlorophyll a/b-protein complex, four minor complexes were revealed, two containing only chlorophyll a and two containing both chlorophyll a and b. The relative distribution of chlorophyll among the resolved chlorophyll-protein complexes and free pigment was found to be similar for all three ploidy levels.     

Effect of Photoperiod on Photosynthate Partitioning and Diurnal Rhythms in Sucrose Phosphate Synthase Activity in Leaves of Soybean (Glycine max L. [Merr.]) and Tobacco (Nicotiana tabacum L.)

Studies were conducted to identify the existence of diurnal rhythms in sucrose phosphate synthase (SPS) activity in leaves of three soybean (Glycine max L. [Merr.]) and two tobacco (Nicotiana tabacum L.) cultivars and the effect of photoperiod (15 versus 7 hours) on carbohydrate partitioning and the rhythm in enzyme activity. Acclimation of all the genotypes tested to a short day (7 hours) photoperiod resulted in increased rates of starch accumulation, whereas rates of translocation, foliar sucrose concentrations, and activities of SPS were decreased relative to plants acclimated to long days (15 hours). Under the long day photoperiod, two of the three soybean cultivars (`Ransom' and `Jupiter') and one of the two tobacco cultivars (`22NF') studied exhibited a significant diurnal rhythm in SPS activity. With the soybean cultivars, acclimation to short days reduced the activity of SPS (leaf fresh weight basis) and tended to dampen the amplitude of the rhythm. With the tobacco cultivars, photoperiod affected the shape of the SPS-activity rhythm. The mean values for SPS activity (calculated from observations made during the light period) were correlated positively with translocation rates and were correlated negatively with starch accumulation rates. Overall, the results support the postulate that SPS activity is closely associated with starch/sucrose levels in leaves, and that acclimation to changes in photoperiod may be associated with changes in the activity of SPS.     

Photosystem II Photosynthetic Unit Sizes from Fluorescence Induction in Leaves : CORRELATION TO PHOTOSYNTHETIC CAPACITY

The use of fluorescence induction measurements in leaves infiltrated with 3-(3,4-dichlorophenyl)-1,1-dimethylurea has been evaluated as a routine method for estimation of the concentration of the reaction centers of photosystem II relative to total chlorophyll in a wide variety of plant species. The procedure is based on a simple theory that takes into account the attenuation of light in passing through the leaf and the linear dependence of the fluorescence induction time from different parts of the leaf on the inverse of the local light intensity. A formula to calculate the reaction center concentration of photosystem II was obtained. The effect of the light attenuation is accounted for by a correction factor which could become practically insignificant by an optimal choice of the excitation and emission wavelengths and the geometry of the photodetector with respect to the sample. Estimation of quantum yields for primary photochemistry and influence of light scattering were considered. The results demonstrate the effect of the above factors under various circumstances and are in agreement, to a first approximation, with the theory.     

Factors Influencing Alanine Aminotransferase Activity in Leaves of Lolium temulentumL.: II. EFFECTS OF GROWTH REGULATORS AND PROTEIN BIOSYNTHESIS INHIBITORS

Effects of kinetin (K), gibberellin A₃ (GA₃), and 2-(chloroethyl)-trimethylammoniumchloride (CCC) on levels of alanine aminotransferase (GPT) andrates of protein synthesis were studied with both intact plantsand isolated leaf segments of Lolium temulentum L. In intactplants CCC stimulated and CA₃ reduced GPT activity, the effectsbsing much greater in 8.h than in 16-h photoporiods. CCC showedmaximum stimulatory effects at 10^–2 M and K at 5 x 10⁵M. No effect of GA₃ could be demonstrated with concentrationsup to 10^–4M. Both K and CCC retarded GPT decline in leafsections, the latter without associated effects upon pigmentbreakdown. Cycloheximide was highly effective in reducing proteinsynthesis in leaf sections. A close correlation between rateof protein synthesis and GPT activity was found over an inhibitorconcentration range from 10^–6 to 10^–4 M. The resultsare discussed in terms of possible methods of in vivo regulationof GPT activity.     

Control of Photosynthetic Sucrose Synthesis by Fructose 2,6-Bisphosphate : II. Partitioning between Sucrose and Starch

The role of fructose 2,6 bisphosphate in partitioning of photosynthate between sucrose and starch has been studied in spinach (Spinacia oleracea U.S. hybrid 424). Spinach leaf material was pretreated to alter the sucrose content, so that the rate of starch synthesis could be varied. The level of fructose 2,6-bisphosphate and other metabolites was then related to the accumulation of sucrose and the rate of starch synthesis. The results show that fructose 2,6-bisphosphate is involved in a sequence of events which provide a fine control of sucrose synthesis so that more photosynthate is diverted into starch in conditions when sucrose has accumulated to high levels in the leaf tissue. (a) As sucrose levels in the leaf rise, there is an accumulation of triose phosphates and hexose phosphates, implying an inhibition of sucrose phosphate synthase and cytosolic fructose 1,6-bisphosphatase. (b) In these conditions, fructose 2,6-bisphosphate increases. (c) The increased fructose 2,6-bisphosphate can be accounted for by the increased fructose 6-phosphate in the leaf. (d) Fructose 2,6-bisphosphate inhibits the cytosolic fructose 1,6-bisphosphatase so more photosynthate is retained in the chloroplast, and converted to starch.     

Partitioning and Utilization of Photosynthate Produced at Different Growth Stages after Anthesis in Soybean (Glycine max L. Merr.): Analysis by Long-term 13C-Labelling Experiments

Yamagata, M., Kouchi, H. and Yoneyama, T. 1987. Partitioningand utilization of photosynthate produced at different growthstages after anthesis in soybean (Glycine max L. Merr.): Analysisby long term ¹³C-labelling experiments.—J. exp. Bot. 38:1247–1259. Soybean (Glycine max L. Merr. var. Akishirome) plants were allowedto assimilate ¹³CO₂ with a constant specific activity for 10h at different growth stages (a total of seven times at aboutone week intervals) after anthesis. The plants were harvestedperiodically until the time of full maturity and the partitioningof ¹³C into individual plant parts was investigated with anemphasis on the contribution of carbon assimilated at differentgrowth stages to the seed formation. Carbon assimilated at the middle to late seed-filling stagecontributed most to the seed production; one day contributionaccounted for 3–4% in total carbon of the seed at fullmaturity. Integrated contribution of carbon assimilated afteranthesis was estimated as 96% of the final seed carbon. An approximationbased on the temporal data of the incorporation of labelledcarbon into the seeds indicates that 77% of the final seed carboncame from direct transfer of current photosynthate from sourceleaves, which occurred within a few days after the photosyntheticfixation, while the rest originated from remobilization of carbonreserved mainly in leaves and stems plus petioles. In comparison with the total carbon accumulation in the seeds,protein carbon in the seeds was relatively more dependent onphotosynthate produced during the early period of reproductivegrowth stage, whereas lipid carbon was more dependent on photosynthateproduced during the later reproductive stage. Key words: Photosynthate partitioning, soybean (Glycine max L. Merr.), ¹³CO₂ assimilation, seed formation     

Studies on Genetic Male-Sterile Soybeans : II. Effect of Nodulation on Photosynthesis and Carbon Partitioning in Leaves

Soybean (Glycine max L. Merr.) germplasm, essentially isogenic except for loci controlling male sterility (ms₁) and nodulation (rj₁), were developed to study the effects of reproductive development and nitrogen source on certain aspects of photosynthesis. Plants were sampled from flowering (77 days after transplanting) until maturity (150 days after transplanting). With all four genotypes, net carbon exchange rates were highest at flowering and declined thereafter. Photosynthetic rates of the sterile genotypes (nodulated and non-nodulated) declined more rapidly than the fertile genotypes, and after 105 days, both sterile genotypes maintained low but relatively constant carbon exchange rates (<3 milligrams CO₂/gram fresh weight per hour). Photosynthetic rates and starch accumulation (difference between afternoon and morning levels) declined with time. The sterile genotypes attained the highest morning starch levels, which reflected reduced starch mobilization. After 92 days, the proportion of photosynthetically fixed carbon that was partitioning into starch (relative leaf starch accumulation) in the sterile genotypes increased dramatically. In contrast, relative leaf starch accumulation in the fertile genotypes remained relatively constant with time. Throughout the test period, all four genotypes maintained leaf sucrose levels between 5 and 15 micromoles glucose equivalents per gram fresh weight.

The activities of sucrose phosphate synthase (SPS) in leaf extracts of the four genotypes declined from 77 to 147 days. Nodulated genotypes tended to maintain higher activities (leaf fresh weight basis) than did the non-nodulated genotypes. In general, relative leaf starch accumulation was correlated negatively with the activity of SPS (normalized with leaf net carbon exchange rate) in leaf extracts for all four genotypes during early reproductive development, and for the fertile genotypes at all sampling dates. In contrast, leaf sucrose content was correlated positively with SPS activity during early reproductive development. These results suggested that a direct relation existed between the activity of SPS and starch/sucrose levels in soybean leaves. However, the interaction between these processes also may be influenced by other factors, particularly when leaf photosynthetic rates and plant demand for assimilates is low, as in the sterile genotypes.

     

RESPONSE OF ANDROPOGON GERARDII (POACEAE) TO FIRE-INDUCED HIGH VS. LOW IRRADIANCE ENVIRONMENTS IN TALLGRASS PRAIRIE: LEAF STRUCTURE AND PHOTOSYNTHETIC PIGMENTS

Photosynthetic pigments and several structural characteristics were measured in leaves of Andropogon gerardii from tallgrass prairie populations in an unburned, low-irradiance site and a burned, high-irradiance site to determine if these species displayed sun/shade differences similar to those documented in forest species. Early in the growing season, leaves of A. gerardii in the low-irradiance, unburned site had significantly lower stomatal density, pore length, and conductance, as well as specific leaf mass and thickness than leaves from the high-irradiance, burned site. Moreover, the chlorophyll a:b ratio, carotenoid content, and bundle sheath-vascular complex area were significantly lower, but chlorophyll content (mass/mass) was greater in leaves in unburned vs. burned sites. These differences are consistent with sun/shade adaptations reported for forest understory plants and may contribute to the low productivity of A. gerardii in unburned tallgrass prairie.     

THE EFFECTS OF RADIATIONS ON BIOLOGICAL SYSTEMS : I. INFLUENCE OF HIGH-FREQUENCY X-RAY RADIATION UPON THE DURATION OF THE PREPUPAL PERIOD OF DROSOPHILAE

The effect of high-frequency x-ray irradiation in prolongation of the larval stage of Drosophila melanogaster has been studied further, and evidence presented of the attainment of a maximum effect followed by a decrease to an almost level plateau in the course curve of average (median) prepupal period (φ) as a function of the period of irradiation (t) under otherwise fixed conditions. The variation of effects of the experimental treatment with age of the larvae at the time of irradiation has been demonstrated in both control and irradiated lots, and a strikingly decreased effect observed when ventilation was not supplied as usual. Means of employment of a living system of this type as an indicator of effectiveness of radiation as in phantom depth or other distributional experiments have been presented and their use illustrated.     

Hemoglobin-digesting Acid Proteinases in Soybean Leaves: CHARACTERISTICS AND CHANGES DURING LEAF MATURATION AND SENESCENCE

Three proteinases which digest hemoglobin rapidly at acid pH (3.5 to 4.5) were identified in crude extracts of soybean (Merr.) leaves and separated by chromatography on DEAE-cellulose. All three enzymes were endopeptidases as judged by the ratio of α-amino-nitrogen plus peptide nitrogen over α-amino-nitrogen in the trichloroacetic acid-soluble portion of hemoglobin digests. Proteinase I did not bind to diethylaminoethyl cellulose and was not inhibited by any of the proteinase inhibitors tested. Proteinase II was partially inhibited by phenylmethylsulfonyl fluoride, N-ethylmaleimide, and p-chloromercuribenzoate. The inhibition by phenyl-methylsulfonyl fluoride can probably be accounted for by the presence of contaminating carboxypeptidase. Proteinase III was the most anionic of the three and required the presence of sulfhydryl reagents to prevent the irreversible loss of activity. All the proteinase preparations digested soy-bean ribulose bisphosphate carboxylase as shown by the disappearance of the large subunit of that protein, when partially digested preparations were subjected to electrophoresis in sodium dodecyl sulfate-polyacrylamide gels. These experiments confirmed that the three proteinases were endopeptidases. All three proteinases were present throughout leaf development; proteinase I predominated in expanding leaves, whereas proteinase III became the predominant enzyme as the leaves matured. Senescence (yellowing) was associated with a decline in the activities of all three proteinases.     

An Endogenous Rhythm in the Rate of CO2 Output of Bryophyllum: II. THE EFFECTS OF LIGHT AND DARKNESS ON THE PHASE AND PERIOD OF THE RHYTHM

The effects of light, darkness, and changes in light intensityon the phase and period of the endogenous rhythm in the rateof CO₂ output of excised leaves of Bryophyllum fedtschenkoihave been examined. The duration, intensity, and wavelength of a short light treatment,and the point in the cycle at which it is administered, determinethe degree of phase shift induced in a rhythm persisting indarkness. When light treatments of 3 and 6 hours' duration,at an intensity of 3,000 lux, are applied between the peaksthe phase is completely reset, the first post-treatment peakoccurring 18–19 hours after the end of the treatment.The degree of phase shift is therefore determined not by theduration of the treatment but by the time at which the treatmentterminates. One hour's illumination has little or no effect.The phase is unaffected when light treatments of up to 5 hours'duration at an intensity of 3,000 lux are applied at the crestof a peak. Over the range 8-3,000 lux the intensity of lightduring a 6-hour treatment applied between the peaks does notaffect the efficiency with which that treatment completely resetsthe phase. At an intensity of 2 lux, however, the phase delayis equal to the duration of the treatment. A 6-hour red-light treatment (850 ergs/cm.²/sec.) applied betweenthe peaks completely resets the phase whereas blue light (10,860ergs/cm.²/sec.) has no effect on the phase but induces a slightprotraction of the period. Moreover, continuous red light inhibitsthe rhythm, which recommences in blue light. A rhythm is induced in illuminated leaves when the light intensityis either gradually or suddenly reduced by at least 80 per cent.Whether a given intensity of illumination inhibits or permitsthe persistence of a rhythm depends upon the light intensityby which it is immediately preceded. A rhythm will persist in illuminated leaves for approximatelyas long as in leaves in darkness and the phase shows no correlationwith time of day. The period is unaffected by the intensityof white light (from 0-500 lux) to which the leaves are subjected.The duration of a short dark treatment, and the point in thecycle at which it is applied, determine the degree of phaseshift induced in a rhythm in illuminated leaves. The phase isreset when 3-, 6-, and 9-hour dark treatments are applied atthe crest of a peak, the amount of phase shift increasing toa maximum with 9 hours' darkness. The phase shift is not equalto the duration of the treatment. The phase is unaffected when3- and 6-hour dark treatments are applied between the peaks. The variation in the sensitivity of the phase of a rhythm persistingin darkness to short light treatments is in the opposite senseto that of a rhythm persisting in light to short dark treatments.The phase of a rhythm in illuminated leaves is completely resetwhen the leaves are transferred to continuous darkness commencingeither at the crest of, or between, the peaks. The results are discussed and compared with those of other authors.     

Transpiration and Photosynthesis in Soybean: EFFECTS OF TEMPERATURE AND VAPOUR PRESSURE DEFICIT

Continuous and simultaneous measurements of CO₂ exchange andtranspiration rates of whole soybean plants were made undercontrasting, controlled environmental conditions for periodsof up to 3 d. Daytime temperatures and vapour pressure deficits(VPD) were 27.5 °C/12 mb; 27.5 °C/5 mb; 22.5 °C/12mb, and 22.5 °C/5 mb. Night temperatures were 5°C lowerthan day temperatures and night VPD was 2.7 mb and 3.5 mb atthe higher and lower temperature respectively. The experimentalconditions were virtually the same as those under which theplants had been grown. Transpiration rates were higher at the higher VPD but were alsoinfluenced by temperature. At 12 mb VPD the rates were 16 percent lower at 22.5 °C than at 27.5 °C. Temperature hadno effect on the transpiration rate at 5 mb VPD. Photosynthesis rates were lower at 5 mb VPD than at 12 mb VPDat both temperatures: the difference was substantially greater(c. 70 per cent) at 22.5 °C. Under all treatments meso-phyllresistance (r'_m) appeared to have a major effect on the photosyntheticrate, and varied more than twofold between treatments. r'_m washighest in plants grown at 22.5 °C/5 mb VPD and lowest at27.5 °C/12 mb VPD.     

Starch Degradation in Spinach Leaves: ISOLATION AND CHARACTERIZATION OF THE AMYLASES AND R-ENZYME OF SPINACH LEAVES

The properties of two amylase activities which differ in their substrate specificity and subcellular location as well as a chloroplast-associated R-enzyme (debranching activity) are reported. An extrachloroplastic amylase is resolved by gel filtration chromatography into two activities of 80,000 and 40,000 daltons. Both extrachloroplastic activities hydrolyze amylopectin and shellfish glycogen and only slowly hydrolyze rabbit liver glycogen, β-limit amylopectin, and amylose. In contrast, the major chloroplastic amylase attacks all of these glucans at comparable rates. Glucan hydrolysis by both the extrachloroplastic and chloroplastic amylase generates not only maltose but appreciable amounts of other oligosaccharides, whereas maltotetraose hydrolysis produces glucose, maltose, and maltotriose. The action patterns displayed by the amylase activities indicate that both are endoamylases, although they lack the typical Ca²⁺ requirement or heat stability of seed endosperm α-amylases. Dithiothreitol, glutathione (oxidized or reduced), ascorbate, dehydroascorbate, and dithiothreitol plus thioredoxin have no effect on either the chloroplastic or extrachloroplastic amylase activities.     

Chilling Damage to Photosynthesis in Young Zea mays: I. EFFECTS OF LIGHT AND TEMPERATURE VARIATION ON PHOTOSYNTHETIC CO2 ASSIMILATION

Carbon dioxide and water vapour exchanges of the second leafof Zea mays in controlled environment cuvettes were measuredin an open gas-exchange system, during and following subjectionto low temperature stress. Photosynthetic CO2 assimilation (Fc)decreased markedly with decrease in leaf temperature so thatFc at 5 °C was c. 7% of Fc at 20 °C. Fc continued todecline if leaf temperature was maintained at 5 °C, andwhen returned to 20 °C the leaf could not regain its previousFc. This chill-induced reduction in the capacity of the leafto assimilate CO2 was proportional to the duration of the chilland increased with water vapour pressure deficit and photonflux density (I_n). Six hours at 5 °C decreased Fc on returnto 20 °C, relative to Fc prior to treatment, by c. 10% indarkness and by c. 50% in a photon flux density approachingfull-sunlight (I_p = 1.5 mmol m^–2 s^–1). The degreeof reduction in Fc following chill treatment showed an almostlinear dependence on both the length and temperature of thechill. Chill treatments resulted in a decrease in both stomataland mesophyll conductances. Examination of the responses ofF_c to light and CO₂ concentration suggested that chill damageto the capacity for CO₂ assimilation resulted from effects onboth the light and CO₂ limited processes within photosynthesis. Key words: Chilling, Photosynthesis, Zea mays, Light-temperature interaction     

Starch Synthesis Studies in Zea mays: II. Molecular Distribution of Radioactivity in Starch

The amylose and amylopectin fractions from kernel starch synthesized shortly after exposure of intact Zea mays L. plants to ¹⁴CO₂ had similar specific radioactivities (counts per min per mg of carbohydrate). In both fractions the radioactivity was distributed throughout the molecules. These data are consistent with a model in which the polysaccharides are synthesized in the matrix of the amyloplast followed by crystallization of the completed molecules onto the starch granule.     

Arginine Metabolism in Developing Soybean Cotyledons : II. Biosynthesis

Tracer kinetic experiments were performed using [ureido-¹⁴C] citrulline, [1-¹⁴C]ornithine, and isotope trapping techniques to determine if arginine is synthesized via the urea cycle in developing cotyledons of Glycine max (L.) Merrill. Excised cotyledons were injected with the ¹⁴C-solution and incubated in sealed vials containing a CO₂ trap. The free and protein amino acids were analyzed using high performance liquid chromatography and arginine-specific enzyme-linked assays. In the ¹⁴C-citrulline feeding experiment argininosuccinate was the most highly labeled compound after 5 minutes and it was the first compound to lose ¹⁴C later in the time course. Carbon-14 was also recovered in free arginine, protein arginine, and CO₂ up to 4 hours after introduction of label. All of the ¹⁴C in free and protein arginine could be accounted for in the C-6 position. Metabolism of ¹⁴C-ornithine resulted in ¹⁴C-incorporation into citrulline and free and protein arginine and the evolution of ¹⁴CO₂. Citrulline was the most highly labeled compound after 15 minutes and was the first compound to reach a steady state level of ¹⁴C. With the addition of 800 nanomoles unlabeled citrulline to the ¹⁴C-ornithine feeding solution citrulline was the only compound labeled after 5 minutes and the steady state level of ¹⁴C-citrulline increased 12-fold. The appearance of ¹⁴C in free arginine and protein arginine was also delayed. In both ¹⁴C-ornithine feedings all of the ¹⁴C in free and protein arginine could be accounted for in the C-1 position. Together, the data support the reaction sequence: ornithine → citrulline → argininosuccinate → arginine → protein arginine.     

Rates of Photosynthesis in Characean Cells: II. PHOTOSYNTHETIC 14CO2 FIXATION AND 14C-BICARBONATE UPTAKE BY CHARACEAN CELLS

Photosynthetic ¹⁴C fixation by Characean cells in solutionsof high pH containing NaH¹⁴CO₃ gave a measure of the abilityof these cells to take up bicarbonate (H¹⁴CO₃^–). Whereascells of Nitella translucens from plants collected and thenstored in the laboratory absorbed bicarbonate at 1–1.5µµmoles cm^–2 sec^–1, rates of 3–8µµmoles cm^–2 sec^–1 were obtained withN. translucens cells from plants grown in the laboratory. Influxesof 5–6 µµmoles cm^–2 sec^–1 wereobtained with Chara australis, 3–8 µµmolescm^–2 sec^–1 with Nitellopsis obtusa, and 1–5µµmoles cm^–2 sec^–1 with Tolypella intricata.It is considered that these influxes represent the activityof a bicarbonate pump, which may be an electrogenic process. In solutions of lower pH, H¹⁴CO₃^– uptake would be maskedby rapid diffusion of ¹⁴CO₂ into the cells: the four Characeanspecies fixed ¹⁴CO₂ at maximum rates of 30–40 µµmolescm^–2 sec^–1 (at 21° C).