Kinetics of C-photosynthate translocation in morning glory vines

The movement of ¹⁴C-photosynthate in morning glory (Ipomea nil Roth, cu. Scarlet O'Hara) vines 2 to 5 meters long was followed by labeling a lone mature leaf with ¹⁴CO₂ and monitoring the arrival rate of tracer at expanding sink leaves on branches along the stem. To a first approximation, the kinetic behavior of the translocation profiles resembled that which would be expected from movement at a single velocity (“plug flow”) without tracer loss from the translocation stream. There was no consistent indication of a velocity gradient along the vine length. The profile moved along the vine as a distinct asymmetrical peak which changes shape only slowly. The spatial distribution of tracer along the vine reasonably matched that predicted on the basis of the arrival kinetics at a sink, assuming plug flow with no tracer loss. These observations are in marked contrast to the kinetic behavior of any mechanism describable by diffusion equations.

However, a progressive change in profile shape (a symmetrical widening) was observed, indicating a range of translocation velocities. A minimum of at least two factors must have contributed to the observed velocity gradient: the exchange of ¹⁴C between sieve elements and companion cells (demonstrated by microautoradiography) and the range of velocities in the several hundred sieve tubes which carried the translocation stream. Possible effects of these two factors on profile spreading were investigated by means of numerical models. The models are necessarily incomplete, due principally to uncertainties about the exchange rate between sieve elements and companion cells and the degree of functional connectivity between sieve tubes of different conductivities. However, most of the observed profile spreading may be reasonably attributed to the combined effects of those two factors.

The mass average velocity of translocation (calculated from the mean times of ¹⁴C arrival at successive sink leaves) was about 75% of the maximum velocity (calculated from the times of initial detection at the same sink leaves), which was usually between 0.6 and 1 cm min⁻¹. Owing to tracer exchange between sieve elements and companion cells, the mass average velocity of tracer in the sieve tubes was probably closer to 86% of the maximum velocity, a figure which agreed with a predicted velocity distribution based on calculated sieve tube conductivities and the size distribution of functional sieve tubes.

     

Rapid Changes in Translocation Patterns in Soybeans following Source-Sink Alterations

The effects of source-sink alterations on the translocation patterns to soybean (“Fiskeby V”) pods were studied using a pod leakage technique. The distribution of assimilates from a source leaf using double pulse experiments was followed at the pods at the source node and the node immediately below. Alterations were made by shading, illuminating, or excising two-thirds of the leaf area of the leaf at the node below. In control experiments both pulses exhibited identical time-course patterns at both nodes. Shading the lower leaf during the first half of the experiment and illuminating during the second reduced the distribution of ¹⁴C-assimilate to the lower node's pods from the source leaf by approximately 30 to 50% while having no effect at the source node. Illuminating the lower leaf during the first half of the experiment followed by excision of two-thirds of that leaf's area and shading increased the import from the source leaf by 4- to 33-fold relative to the control while reducing the distribution to the source node by up to 40%. The change in distribution pattern took place in less than 30 minutes with no apparent change in the source leaf net photosynthesis or in the rate of movement to the pods. The results indicate that any alterations in the source-sink balance will quickly produce a change in the distribution patterns to the pods.     

Source pool kinetics for C-photosynthate translocation in morning glory and soybean

The kinetic behavior of translocation profiles indicates that their shape is determined largely by the rate at which tracer enters the sieve tubes in the source leaf. Confirmation of this relationship was sought by investigating the kinetics of ¹⁴C in the immediate source pool for translocated sucrose in soybean (Glycine max L., cv. Bragg) and morning glory (Ipomea nil Roth, cv. Scarlet O'Hara) leaves. Quantitative microautoradiography was used to follow the water-soluble ¹⁴C contents of the companion cells in minor veins after pulse-labeling with ¹⁴CO₂. In both morning glory and soybean, the observed kinetics in the companion cells matched reasonably well those expected from the shape of the translocation profiles.

Marked compartmentation of sucrose was evident in soybean leaves in that the specific radioactivity of total leaf sucrose was greatest immediately after labeling and quickly declined, whereas labeling in the companion cells was low at first and did not reach a maximum for about 35 minutes. In morning glory leaves, the kinetics of sucrose specific radioactivity and of companion cell-labeling more closely paralleled one another.

     

Apetiolar photosynthate translocation

Apetiolar transport of photosynthate ⁻¹⁴C has been studied by feeding of ¹⁴CO₂ to soybean petioles. Translocation occurs in the absence of leaves, but both the rate and velocity are diminished. The effect of root excision is not as profound as that of leaves. It appears, in some instances, to inhibit transport partially, so that accumulation of photosynthate develops, giving a steeper isotopic gradient. The effect of leaf darkening is to diminish its uptake of photosynthate from the petiole, possibly as a result of decreased transpiration in the lowered temperature of the darkened leaf. The data suggest that neither mass flow nor active transport provide an adequate basis for normal photosynthate transport but that the leaves provide a direct force requiring structural continuity, or a translocation carrier.     

Kinetics of C-14 Translocation in Soybean: II. Kinetics in the Leaf

The kinetics of ¹⁴C-assimilates in the soybean leaf were studied in pulse labeling and steady state labeling experiments. ¹⁴C-Sucrose apparently served as the ultimate source, at least, of translocated ¹⁴C-sucrose. However, since the specific activity of leaf sucrose reached a maximum within 5 minutes after pulse labeling, whereas that of exported sucrose did not reach a maximum until at least 20 minutes, it appeared that there were two sucrose compartments in the leaf. A possible physical basis for the two compartments may be the mesophyll (a photosynthetic compartment) and a specialized “paraveinal mesophyll” (a nonphotosynthetic compartment), through which photosynthate must pass on its way to the veins.     

Kinetics of C-14 translocation in soybean: I. Kinetics in the stem

A kinetic study was made of the translocation of ¹⁴C-photosynthate through soybean stems following pulse labeling and during steady state labeling of the first trifoliolate leaf. The translocation profile proceeded down the stem with little or no change in shape. Following pulse labeling, sucrose accounted for 90 to 95% of the radioactivity in the stem at all times up to 2 hours, at which time less than 3% of the activity was in an insoluble form. Kinetic data on the relative specific activities of sucrose in the leaf and petiole indicated that two-thirds of the petiolar sucrose was in the translocation stream and the remaining one-third was in a stationary pool which slowly accumulated sucrose from the translocation stream. With this assumption, the rate of sucrose efflux from the leaf was calculated to be 22 micrograms per minute, which was equivalent to a sucrose mass flux in the sieve tubes of 20 grams per square centimeter per hour.     

Influence of assimilate demand on photosynthesis, diffusive resistances, translocation, and carbohydrate levels of soybean leaves

Rates of net photosynthesis and translocation, CO₂ diffusive resistances, levels of carbohydrates, total protein, chlorophyll, and inorganic phosphate, and ribulose 1,5-diphosphate carboxylase activity were measured in soybean (Glycine max L. Merrill) leaves to ascertain the effect of altered assimilate demand. To increase assimilate demand, the pods, stems, and all but one leaf (the “source leaf”) of potted plants were completely shaded for 6 or 8 days and the responses of the illuminated source leaf were monitored. Rate of net photosynthesis in the source leaf of the shaded plants was found to increase curvilinearly to a maximum on the 8th day. The source leaf of the control plants (no sink shading) maintained a constant photosynthetic rate during this period. Vapor-phase resistance to CO₂ diffusion did not vary with treatment, but mesophyll (liquid phase) resistance was significantly lower in the source leaf of the shaded plants.     

Alterations in source-sink patterns by modifications of source strength

Bean plants, trimmed to a simplified “double source, double sink” translocation system (the paired primary leaves serving as the double source and the paired lateral leaflets of the immature first trifoliate leaf as the double sink) were used to study the magnitude and short-term time course of change in the allocation ratio (partition ratio) of assimilates translocated from the labeled primary leaf to its respective “near” and “far leaflet” sinks in response to an increase or decrease in the source strength of the opposite primary leaf (the “control” leaf). If the rates of net photosynthesis in the two primary leaves were similar, assimilates from the labeled source leaf partitioned to the leaflet sinks in the ratio of 5:1 or higher, the dominant sink being the leaflet “nearer” to the labeled source leaf. If the rate of net photosynthesis in the control leaf was increased substantially above that of the labeled source leaf, the rate of translocation from the labeled source to either the near leaflet sink or far leaflet sink remained unaffected, despite, presumably, a higher translocation rate from the control leaf, and hence a higher phloem pressure gradient (or increased cross-sectional area) in the transport pathway from the control leaf to the leaflet sinks. If the control leaf was excised, thus reducing the source leaf area by about a half, the translocation rate from the remaining source leaf rapidly doubled, the partition ratio becoming equal to unity. If the control leaf was darkened, the partition ratio adjusted to an intermediate value. Although export rates from the labeled source leaf were increased either by excising or darkening the control leaf, the rate of net photosynthesis in the labeled leaf remained constant.     

Observations of Hydrogen and Oxygen Isotopes in Leaf Water Confirm the Craig-Gordon Model under Wide-Ranging Environmental Conditions

The Craig-Gordon evaporative enrichment model of the hydrogen (δD) and oxygen (δ¹⁸O) isotopes of water was tested in a controlled-environment gas exchange cuvette over a wide range (400‰ δD and 40‰ δ¹⁸O) of leaf waters. (Throughout this paper we use the term “leaf water” to describe the site of evaporation, which should not be confused with “bulk leaf water” a term used exclusively for uncorrected measurements obtained from whole leaf water extractions.) Regardless of how the isotopic composition of leaf water was achieved (i.e. by changes in source water, atmospheric vapor δD or δ¹⁸O, vapor pressure gradients, or combinations of all three), a modified version of the Craig-Gordon model was shown to be sound in its ability to predict the δD and δ¹⁸O values of water at the site of evaporation. The isotopic composition of atmospheric vapor was shown to have profound effects on the δD and δ¹⁸O of leaf water and its influence was dependent on vapor pressure gradients. These results have implications for conditions in which the isotopic composition of atmospheric vapor is not in equilibrium with source water, such as experimental systems that grow plants under isotopically enriched water regimes. The assumptions of steady state were also tested and found not to be a major limitation for the utilization of the leaf water model under relatively stable environmental conditions. After a major perturbation in the δD and δ¹⁸O of atmospheric vapor, the leaf reached steady state in approximately 2 h, depending on vapor pressure gradients. Following a step change in source water, the leaf achieved steady state in 24 h, with the vast majority of changes occurring in the first 3 h. Therefore, the Craig-Gordon model is a useful tool for understanding the environmental factors that influence the hydrogen and oxygen isotopic composition of leaf water as well as the organic matter derived from leaf water.     

Kinetic characteristics of photoassimilate translocation in Alaria esculenta (Laminariales,Phaeophyceae)

The kinetics of translocation of ¹⁴C-labeled photoassimilate were studied in the kelp, Alaria esculenta (L.) Grev., using a Geiger-Müller detector-probe to measure radioactivity in the source and sink regions of dumbbell-shaped explants cut from blades. Rapid tracer efflux from the source occurred for 4 days following a pulse of [¹⁴C]bicarbonate, with 40–60% of the initial activity remaining in the source after 10–14 days. Portions of source and sink tissue were analysed for distribution of radioactivity in mannitol, amino-acid, organic-acid and insoluble fractions. About 75% of the radioactivity in both source and sink at the end of the experiments was in soluble organic matter. The translocation velocity of the moving solute front (1.0-1.6 cm·h^-1) was derived from time-course profiles of tracer arriving in Alaria sinks. Relative rates of translocation, calculated from these profiles, yielded skewed curves, with maximum rates of import by the sink occurring 72–96 h after the source was pulsed.     

A Method for Calculating Sucrose Synthesis Rates throughout a Light Period in Sugar Beet Leaves

Sucrose synthesis rate in an exporting sugar beet (Beta vulgaris L.) leaf was calculated from simultaneous measurements of export and changes in leaf sucrose level. The amount of recently fixed carbon exported was determined from net carbon assimilated minus the tracer carbon accumulated in the leaf. The relative amount of ¹⁴C accumulated in the leaf supplied with ¹⁴CO₂ throughout an entire light period was recorded continuously with a Geiger-Mueller detector. To produce a continuous time course for tracer carbon accumulated in the leaf during the light period, the latter curve was superimposed on values for tracer carbon accumulated in leaves sampled at hourly intervals. Validity of the method requires that nearly all of the carbon that is exported be sucrose and that nearly all of the sucrose that is synthesized be either exported or accumulated as sucrose in the exporting leaves. These conditions appeared to be fulfilled in the situations where the method was applied. The method was used to study the effect of increasing atmospheric CO₂ concentration on the rate of sucrose synthesis. Further, the method can be used in conjunction with the gathering of other data such as gas exchange, metabolite levels, and enzyme activities in a set of leaves of a similar age on the same plant. This assemblage of data was found to be useful for understanding how rates of photosynthesis, sucrose synthesis, and translocation are regulated in relation to each other in an intact plant.     

The influence of externally supplied sucrose on phloem transport in the maize leaf strip

Sucrose (2,5–1000 mmol l^–1), labeled with [¹⁴C]sucrose, was taken up by the xylem when supplied to one end of a 30-cm-long leaf strip of Zea mays L. cv. Prior. The sugar was loaded into the phloem and transported to the opposite end, which was immersed in diluted Hoagland's nutrient solution. When the Hoagland's solution at the opposite end was replaced by unlabeled sucrose solution of the same molarity as the labeled one, the two solutions met near the middle of the leaf strip, as indicated by radioautographs. In the dark, translocation of ¹⁴C-labeled assimilates was always directed away from the site of sucrose application, its distance depending on sugar concentration and translocation time. When sucrose was applied to both ends of the leaf strip, translocation of ¹⁴C-labeled assimilates was directed toward the lower sugar concentration. In the light, transport of ¹⁴-C-labeled assimilates can be directed (1) toward the morphological base of the leaf strip only (light effect), (2) toward the base and away from the site of sucrose application (light and sucrose effect), or (3) away from the site of sucrose application independent of the (basipetal or acropetal) direction (sucrose effect). The strength of a sink, represented by the darkened half of a leaf strip, can be reduced by applying sucrose (at least 25 mmol l^–1) to the darkened end of the leaf strip. However, equimolar sucrose solutions applied to both ends do not affect the strength of the dark sink. Only above 75 mmol l^–1 sucrose was the sink effect of the darnened part of the leaf strip reduced. Presumably, increasing the sucrose concentration replenishes the leaf tissue more rapidly, and photosynthates from the illuminated part of the leaf strip are imported to a lesser extent by the dark sink.Supported by Deutsche Forschungsgemeinschaft     

Independent translocation of 14C-labelled assimilates and of the floral stimulus in Lolium temulentum

Summary It is widely accepted that the floral stimulus produced in leaves is carried to the shoot apex passively in the phloem with the assimilate stream. Three kinds of evidence presented here suggest that the floral stimulus moves independently of the assimilates.Simultaneous determination of the velocities of translocation out of the seventh leaf blade, in comparable plants under the same conditions, yielded estimates of 1–2.4 cm/hr for the floral stimulus, and 77–105 cm/hr for ¹⁴C-labelled assimilates.The effect of the size of the seventh leaf on its ability to export assimilates or to initiate flowering was quite different. Leaves with only 14–26% of their final blade area emerged exported little assimilate, yet were highly active in inducing flowering.The effect of DCMU applications at a range of concentrations on the translocation of assimilates was quite different from their effect on the flowering response.     

Influence of cobalt on soybean hypocotyl growth and its ethylene evolution

Development of dark-grown “Clark” soybean (Glycine max [L.] Merr.) seedlings is abnormal at 25 C but normal at 20 and 30 C. At 25 C, hypocotyls swell and fail to elongate normally; lateral root formation and seedling ethylene evolution are enhanced.

Co²⁺ promoted hypocotyl elongation of etiolated “Clark” soybean seedlings by 28% when grown at 25 C. The same growth-promoting concentration reduced hypocotyl thickness and primary root elongation by 28 and 43%, respectively. Co²⁺ inhibited ethylene production both of intact seedlings and of apical 1-centimeter hypocotyl segments with attached epicotyls and cotyledons by 65 and 60%, respectively. These results suggest that Co²⁺ exerts its effects on the hypocotyl growth by inhibiting ethylene production, and also confirm our previous conclusion that abnormal ethylene production at 25 C is responsible for the inhibition of hypocotyl elongation and for its swelling.

     

Interactive effects of light,leaf temperature,CO2 and O2 on photosynthesis in soybean

A biochemical model of C ₃photosynthesis has been developed by G.D. Farquhar et al. (1980, Planta 149, 78–90) based on Michaelis-Menten kinetics of ribulose-1,5-bisphosphate (RuBP) carboxylase-oxygenase, with a potential RuBP limitation imposed via the Calvin cycle and rates of electron transport. The model presented here is slightly modified so that parameters may be estimated from whole-leaf gas-exchange measurements. Carbon-dioxide response curves of net photosynthesis obtained using soybean plants (Glycine max (L.) Merr.) at four partial pressures of oxygen and five leaf temperatures are presented, and a method for estimating the kinetic parameters of RuBP carboxylase-oxygenase, as manifested in vivo, is discussed. The kinetic parameters so obtained compare well with kinetic parameters obtained in vitro, and the model fits to the measured data give r ²values ranging from 0.87 to 0.98. In addition, equations developed by J.D. Tenhunen et al. (1976, Oecologia 26, 89–100, 101–109) to describe the light and temperature responses of measured CO₂-saturated photosynthetic rates are applied to data collected on soybean. Combining these equations with those describing the kinetics of RuBP carboxylase-oxygenase allows one to model successfully the interactive effects of incident irradiance, leaf temperature, CO₂ and O₂ on whole-leaf photosynthesis. This analytical model may become a useful tool for plant ecologists interested in comparing photosynthetic responses of different C₃ plants or of a single species grown in contrasting environments.Abbreviations PCO photorespiratory carbon oxidation - PCR photosynthetic carbon reduction - PPFD photosynthetic photon-flux density - RuBP ribulose bisphosphate     

Comparative photorespiration in Amaranthus,soybean and corn

Summary A tracer technique was used to measure photorespiration in Amaranthus lividus, soybean and corn. Under a light intensity of 40 Wm^-2 (400–700 nm) efflux of tracer carbon dioxide from Amaranthus into air was comparable to that from soybean over a 30-min period and 10 times that from corn. Initial rates of efflux of tracer into air from Amaranthus were higher than from soybean and 9 times that from corn. Efflux of CO₂ from Amaranthus over 30 min in 120 Wm^-2 was only 5 times that of corn and the initial rate was only one third that of soybean. Though total efflux from soybean was similar at the two light intensities, the initial rate was slightly higher under 120 Wm^-2. For Amaranthus and soybean, pure oxygen doubled total efflux of CO₂ and substantially increased the initial rate compared with CO₂-free air whereas there was no effect on corn. A comparison of the light and dark curves suggests that light and dark respiration had different substrates. The results are interpreted in terms of the recycling of photorespiratory CO₂.     

Use of positron-emitting tracer imaging system for measuring the effect of salinity on temporal and spatial distribution of C tracer and coupling between source and sink organs

Salinity stress affects photosynthate partitioning between sources and sinks of plants, but how it affects these systems is less well understood. Because sources and sinks are closely tied, any adverse effect under suboptimal conditions on one of these is often misinterpreted for an effect on the other. Carbon partitioning is indispensable for stress resistance and good plant growth. In the present study, carbon partitioning in tomato plants (Lycopersicon esculentum L. cv. Momotarou) in a saline (NaCl) environment was studied by feeding radioactive ¹¹C and stable ¹³C isotopes. Pulse-chases were conducted to measure the spatial and temporal distribution of ¹³C. ¹³C was measured by a standard conventional technique, but ¹¹C distribution was monitored using a positron-emitting tracer imaging system (PETIS). Salt stress resulted in reduced carbon translocation toward roots. The majority of the photosynthate accumulated in the leaf. We also observed that the reduction in translocation of carbon occurred well before the salt stress symptoms of reduced photosynthesis and reduced plant growth in salt-exposed plants. The effect on sink activity was also shown by a decrease in stem diameter. In addition, PETIS analysis of ¹¹C translocation indicated that carbon translocation to roots was inhibited under salt conditions without a direct effect on leaf Na accumulation or osmotic stress. These results suggest that NaCl has direct effects on plants, inhibiting carbon partitioning within a few hours of salt exposure without inhibition of source activity.     

The influence of calcium deficiency on the translocation of photosynthetically fixed 14C in soybeans

Summary Soybean (Glycine max (L.) Merr.) plants were grown at two levels of Ca (0.05 and 2.5 mM Ca) with and without an inorganic source of N in a growth chamber and in the greenhouse. The fourth leaf from the top of the plant was labeled with ¹⁴C, and the distribution of ¹⁴C was measured 24 hours after labeling. The translocation of ¹⁴C out of the labeled leaf was decreased at the low Ca level in both experiments. This decrease occurred before there were any visual Ca deficiency symptoms and before the low Ca level had any effect on dry weight accumulation. The effect of the low Ca level on translocation was the same for plants exposed to an inorganic N source and those that were nodulated. The reduced rate of translocation from the labeled leaf had no measurable effect on the nodule activity as measured by the acetylene reduction method. The data demonstrate the importance of an adequate supply of Ca to the soybean plant for maximum translocation of carbohydrates from the leaves.This paper (76-3-110) is published with the approval of the Director of the Kentucky Agric. Exp. Stn.This paper (76-3-110) is published with the approval of the Director of the Kentucky Agric. Exp. Stn.     

Phosphorus absorption and utilization efficiency of pigeon pea (Cajanus cajan (L) Millsp.) in relation to dry matter production and dinitrogen fixation

The effect of P supply on absorption and utilization efficiency of P in relation to dry matter production and dinitrogen fixation was examined in 8 pigeon pea cultivars with different growth duration and a soybean cultivar under field conditions. In all the pigeon pea cultivars, the maximum whole plant dry weight was obtained in a P-deficient soil at 100 kg P ha⁻¹ application. The short duration cultivars had smaller whole plant dry weights at low P rates (5 and 25 kg P ha⁻¹) and poor response to P application compared with the medium and long duration cultivars. Increasing the P application rate significantly increased dinitrogen fixation in all the cultivars. At the low P rates, the total nodule activity (TNA) was lower in the short than in the medium and the long duration cultivars. However, at 200 kg P ha⁻¹ application, dinitrogen fixation did not vary among these cultivars except for one short duration cultivar whichregistered very low values. Dry matter production and dinitrogen fixation are strongly controlled by P absorption ability rather than P utilization efficiency. The low absorption ability of the short duration cultivars is mainly due to poor root development. The high P concentrations in the nodules of all the cultivars suggest that nodules have advantage over host plant interms of P distribution under P deficient conditions. Our results suggest that P plays an important role in dinitrogen fixation through an effective translocation of P to the leaf. Thus when P supply is limited, efficient cultivars obtained reasonably high yield through an effective translocation of the absorbed P to the leaf.     

The number of chlorophyll-less spots and the primary leaf area as criteria of radiation damage in soybean (Glycine max merill)

The number of chlorophyll-less spots occurring on the primary leaves as well as the primary leaf size were investigated in two soybean cultivars, differing genetically in radiosensitivity, after irradiation of seed with ⁶⁰Co γ-rays. A high correlation was found between increasing number of spots, decreasing leaf size and seedling growth inhibition.The number of spots can be used to monitor radiation effects over the small dose range where the growth inhibition is not pronounced. Primary leaf size can be used as a convenient criterion of seedling growth inhibition. Possible causes of leaf spotting are discussed.