Reduced Cytosolic Fructose-1,6-Bisphosphatase Activity Leads to Loss of O(2) Sensitivity in a Flaveria linearis Mutant

The mutant plant of Flaveria linearis characterized by Brown et al. (Plant Physiol. 81: 212-215) was studied to determine the cause of the reduced sensitivity to O₂. Analysis of CO₂ assimilation metabolites of freeze clamped leaves revealed that both 3-phosphoglycerate and ribulose 1,5-bisphosphate were high in the mutant plant relative to F. linearis with normal O₂ sensitivity. The k_cat of ribulose-1,5-bisphosphate carboxylase (RuBPCase) was equal in all plant material tested (range 18-22 s⁻¹) indicating that no tight binding inhibitor was present. The degree of RuBPCase carbamylation was reduced in the mutant plant relative to the wild-type plant. Since 3-phosphoglycerate was high in the mutant plant and photosynthesis did not exhibit properties associated with RuBPCase limitations, we believe that the decarbamylation of RuBPCase was a consequence of another lesion in photosynthesis. Fructose 1,6-bisphosphate and its precursors, such as the triose phosphates, were in high concentration in the mutant plant relative to the wild type. The concentrations of the product of the fructose 1,6-bisphosphatase reaction, fructose 6-phosphate, and its isomer, glucose 6-phosphate, were the same in both plants. We found that the mutant plant had up to 75% less cytosolic fructose 1,6-bisphosphatase activity than the wild type but comparable levels of stromal fructose 1,6-bisphosphatase. We conclude that the reduced fructose-1,6-bisphosphatase activity restricts the mutant plant's capacity for sucrose synthesis and this leads to reduced or reversed O₂ sensitivity.     

Oxygen Stimulation of Apparent Photosynthesis in Flaveria linearis

A plant was found in the C₃-C₄ intermediate species, Flaveria linearis, in which apparent photosynthesis is stimulated by atmospheric O₂ concentrations. A survey of 44 selfed progeny of the plant showed that the O₂ stimulation of apparent photosynthesis was passed on to the progeny. When leaves equilibrated at 210 milliliters per liter O₂ were transferred to 20 milliliters per liter O₂ apparent photosynthesis was initially stimulated, but gradually declined so that at 30 to 40 minutes the rate was only about 80 to 85% of that at 210 milliliters per liter O₂. Switching from 20 to 210 milliliters per liter caused the opposite transition in apparent photosynthesis. All other plants of F. linearis reached steady rates within 5 minutes after switching O₂ that were 20 to 24% lower in 210 than in 20 milliliters per liter O₂. At low intercellular CO₂ concentrations and low irradiances, O₂ inhibition of apparent photosynthesis of the aberrant plant was similar to that in normal plants, but at an irradiance of 2 millimoles quanta per square meter per second and near 300 microliters per liter CO₂ apparent photosynthesis was consistently higher at 210 than at 20 milliliters per liter O₂. In morphology and leaf anatomy, the aberrant plant is like the normal plants in F. linearis. The stimulation of apparent photosynthesis at air levels of O₂ in the aberrant plant is similar to other literature reports on observations with C₃ plants at high CO₂ concentrations, high irradiance and/or low temperatures, and may be related to limitation of photosynthesis by triose phosphate utilization.     

Carbon Partitioning in a Flaveria linearis Mutant with Reduced Cytosolic Fructose Bisphosphatase

Oxygen sensitivity and partitioning of carbon was measured in a mutant line of Flaveria linearis that lacks most of the cytosolic fructose-1,6-bisphosphatase found in wild-type lines. Photosynthesis of leaves of the mutant line was nearly insensitive to O₂, as found before. The mutant plants partitioned 2.5 times less carbon into sucrose than the wild type in a pulse chase experiment, with the extra carbon going mainly to starch but also to amino acids. From 10 to 50 min postlabeling, radioactivity chased out of the amino acid fraction to starch in both lines. In the middle of the light period, starch grains were larger in the mutant than in the wild type and covered 30% of the chloroplast area as seen with an electron microscope. Starch grains were found in both mesophyll and bundle sheath chloroplasts in both lines in these C₃-C₄ intermediate plants. At the end of the dark period, the starch levels were considerably reduced from what they were in the middle of the light in both lines. The concentration of sucrose was higher in the mutant line despite the lack of cytosolic fructose-1,6-bisphosphatase. The amino acid fraction accounted for about 30% of all label following a 10-min chase period. In the mutant line, most of the label was in the glycine + serine fraction, with 10% in the alanine fraction. In wild-type leaves, 35% of the label in amino acids was in alanine. These results indicate that this mutant survives the reduced cytosolic fructose-1,6-bisphosphatase activity by partitioning more carbon to starch and less to sucrose during the day and remobilizing the excess starch at night. However, these results raise two other questions about this mutant. First, why is the sucrose concentration high in a plant that partitions less carbon to sucrose, and second, why is alanine heavily labeled in the wild-type plants but not in the mutant plants?     

Photosynthetic Characteristics of Segregates from Hybrids between Flaveria brownii (C4 Like) and Flaveria linearis (C3-C4)

Characteristics related to C4 photosynthesis were studied in reciprocal F1 hybrids and F2 plants from Flaveria brownii (C4 like) and Flaveria linearis (C3-C4). The reciprocal F1 plants differed in 13C/12C ratios of leaves and the percentage of 14C initially incorporated into C4 acids, being more like the pollen parents in these traits. They did not differ in apparent photosynthesis or in O2 inhibition of apparent photosynthesis and differed only slightly in CO2 compensation concentration at 175 [mu]mol quanta m-2 s-1 and 400 mL L-1 O2. The 13C/12C ratios of 78 F2 progeny from the two F1 plants exhibited a normal distribution centered between those of the parents, with a few values slightly higher and lower than the parents. Apparent photosynthesis at 130 [mu]L L-1 CO2 and inhibition of photosynthesis by O2 was nearly normally distributed in the F2 population, but no values for F2 plants approached those for F. brownii (15.4 [mu]mol m-2 s-1 and 7.8%, respectively). Distribution of the CO2 compensation concentration measured at 1000 [mu]mol quanta m-2 s-1 and 400 mL L-1 of O2 in the F2 population was skewed toward F. brownii with 72% of the progeny having values <9 [mu]L of CO2 L-1 compared to 1.5 and 27.2 [mu]L L-1 for F. brownii and F. linearis, respectively. Correlations among traits of F2 plants were low (coefficients of 0.30 to -0.49), indicating that the C4- related traits are not closely linked in segregating populations. Plants in the F2 population selected for high or low apparent photosynthesis at 130 [mu]L of CO2 L-1 (six each) did not rank consistently high or low for 13C/12C ratios, O2 inhibition of apparent photosynthesis, CO2 compensation concentration, or activities of phosphoenolpyruvate carboxylase or NADP-malic enzyme. This study confirms results of earlier work that indicates independent segregation of C4 traits and also shows that the C4-like parental type can be recovered, at least for some characteristics (13C/12C ratio), in segregating populations. Recovery of fully functional C4 plants awaits further experimentation with C4 x C3 or C4 x C3-C4 hybrid plants that produce fertile progeny.     

Photosynthetic Characteristics of C(3)-C(4) Intermediate Flaveria Species : III. Reduction of Photorespiration by a Limited C(4) Pathway of Photosynthesis in Flaveria ramosissima

The initial products of photosynthesis by the C₃ species Flaveria cronquistii, the C₄ species F. trinervia, and the C₃-C₄ intermediate species F. ramosissima were determined using a pulse-chase technique with ¹⁴CO₂-¹²CO₂. The intermediate species F. ramosissima incorporated at least 42% of the total soluble ¹⁴C fixed into malate and aspartate after 10 seconds of photosynthesis in ¹⁴CO₂, as compared with 90% for the C₄ species F. trinervia and 5% for the C₃ species F. cronquistii. In both F. ramosissima and F. trinervia, turnover of labeled malate and aspartate occurred during a chase period in ¹²CO₂, although the rate of turnover was slower in the intermediate species. Relative to F. cronquistii, F. ramosissima showed a reduced incorporation of radioactivity into serine and glycine during the pulse period. These results indicate that a functional C₄ pathway of photosynthesis is operating in F. ramosissima which can account for its reduced level of photorespiration, and that this species is a true biochemical intermediate between C₃ and C₄ plants.     

Evolution of C4 Photosynthesis in the Genus Flaveria: Establishment of a Photorespiratory CO2 Pump

C₄ photosynthesis is nature’s most efficient answer to the dual activity of ribulose-1,5-bisphosphate carboxylase/oxygenase and the resulting loss of CO₂ by photorespiration. Gly decarboxylase (GDC) is the key component of photorespiratory CO₂ release in plants and is active in all photosynthetic tissues of C₃ plants, but only in the bundle sheath cells of C₄ plants. The restriction of GDC to the bundle sheath is assumed to be an essential and early step in the evolution of C₄ photosynthesis, leading to a photorespiratory CO₂ concentrating mechanism. In this study, we analyzed how the P-protein of GDC (GLDP) became restricted to the bundle sheath during the transition from C₃ to C₄ photosynthesis in the genus Flaveria. We found that C₃ Flaveria species already contain a bundle sheath–expressed GLDP gene in addition to a ubiquitously expressed second gene, which became a pseudogene in C₄ Flaveria species. Analyses of C₃-C₄ intermediate Flaveria species revealed that the photorespiratory CO₂ pump was not established in one single step, but gradually. The knowledge gained by this study sheds light on the early steps in C₄ evolution.     

Photosynthetic Plasticity in Flaveria brownii: Growth Irradiance and the Expression of C(4) Photosynthesis

Photosynthesis was examined in leaves of Flaveria brownii A. M. Powell, grown under either 14% or 100% full sunlight. In leaves of high light grown plants, the CO₂ compensation point and the inhibition of photosynthesis by 21% O₂ were significantly lower, while activities of ribulose 1,5-bisphosphate carboxylase/oxygenase and various C₄ cycle enzymes were considerably higher than those in leaves grown in low light. Both the CO₂ compensation point and the degree of O₂ inhibition of apparent photosynthesis were relatively insensitive to the light intensity used during measurements with plants from either growth conditions. Partitioning of atmospheric CO₂ between Rubisco of the C₃ pathway and phosphoenolpyruvate carboxylase of the C₄ cycle was determined by exposing leaves to ¹⁴CO₂ for 3 to 16 seconds, and extrapolating the labeling curves of initial products to zero time. Results indicated that ~94% of the CO₂ was fixed by the C₄ cycle in high light grown plants, versus ~78% in low light grown plants. Thus, growth of F. brownii in high light increased the expressed level of C₄ photosynthesis. Consistent with the carbon partitioning patterns, photosynthetic enzyme activities (on a chlorophyll basis) in protoplasts from leaves of high light grown plants showed a more C₄-like pattern of compartmentation. Pyruvate, Pi dikinase and phosphoenolpyruvate carboxylase were more enriched in the mesophyll cells, while NADP-malic enzyme and ribulose 1,5-bisphosphate carboxylase/oxygenase were relatively more abundant in the bundle sheath cells of high light than of low light grown plants. Thus, these results indicate that F. brownii has plasticity in its utilization of different pathways of carbon assimilation, depending on the light conditions during growth.     

Photosynthesis of F(1) Hybrids between C(4) and C(3)-C(4) Species of Flaveria

Photosynthetic characteristics were studied in several F₁ hybrids between C₄ and C₃-C₄ species of Flaveria. Stable carbon isotope ratios, O₂ inhibition of apparent photosynthesis, and phosphoenolpyruvate carboxylase activities in the hybrids were similar to the means for the parents. Values of CO₂ compensation concentrations were nearer to those of the C₄ parent and apparent photosynthesis was below that of both parents, being only 60 and 74% of that of the lowest (C₃-C₄) parent in two experiments. Reductions of CO₂ compensation concentration and O₂ inhibition of apparent photosynthesis as well as increases in carbon isotope ratios and phosphoenolpyruvate carboxylase activities compared to values in C₃-C₄ species suggest transfer of a limited degree of C₄ photosynthesis to the F₁ hybrids. However, the lower apparent photosynthesis of the hybrids suggests that transfer of C₄ characteristics to non-C₄ species is detrimental unless characteristics associated with C₄ photosynthesis are fully developed. There was a highly significant negative correlation (r = −0.90) between CO₂ compensation concentration and the logarithm of phosphoenolpyruvate carboxylase activity in the parents and hybrids, suggesting involvement of this enzyme in controlling the CO₂ compensation concentration. Although bundle-sheath cells were more developed in leaves of hybrids than in C₃-C₄ parents, they appeared to contain lower quantities of organelles than those of the C₄ parent. Reduced quantities of organelles in bundle-sheath cells could indicate incomplete compartmentation of partial pathways of the C₄ cycle in the hybrids. This may mean that the reduction of CO₂ compensation and O₂ inhibition of apparent photosynthesis relative to the C₃-C₄ parents is less dependent on fully developed Kranz anatomy than is increased apparent photosynthesis.     

Effects of CO(2) and O(2) on Photosynthesis and Growth of Autotrophic Tobacco Callus

Gas exchange measurements were made on plants from two natural populations differing in salt tolerance of Andropogon glomeratus, a C₄ nonhalophyte, to examine the effect of salinity on components responsible for differences in photosynthetic capacity. Net CO₂ uptake and stomatal conductance decreased with increasing salinity in both populations, but to a greater extent in the inland (nontolerant) population. The intercellular CO₂ concentrations increased with increasing salinity in the inland population, but decreased in the marsh (tolerant) population. Water use efficiency decreased as salinity increased in the inland population, and remained unchanged in the marsh population. Carboxylation efficiency decreased and CO₂ compensation points increased with increasing salinity in both populations, but to a lesser extent in the marsh population. Carboxylation efficiencies were higher with 2% relative to 21% atmospheric O₂ in salt stressed plants, suggesting that a decrease in the carboxylation:oxygenation ratio of ribulose 1,5-bisphosphate carboxylase/oxygenase was partly responsible for the decrease in photosynthetic capacity. Populational differences in photosynthetic capacity were the result of greater salinity-induced changes in carboxylation efficiency in the inland population, and not due to differences in the stomatal limitation to CO₂ diffusion.     

Degree of C(4) Photosynthesis in C(4) and C(3)-C(4)Flaveria Species and Their Hybrids : II. Inhibition of Apparent Photosynthesis by a Phosphoenolpyruvate Carboxylase Inhibitor

Hybrids have been made between species of Flaveria exhibiting varying levels of C₄ photosynthesis. The degree of C₄ photosynthesis expressed in four interspecific hybrids (Flaveria trinervia [C₄] × F. linearis [C₃-C₄], F. brownii [C₄-like] × F. linearis, and two three-species hybrids from F. trinervia × [F. brownii × F. linearis]) was estimated by inhibiting phosphoenolpyruvate carboxylase in vivo with 3,3-dichloro-2-dihydroxyphosphinoylmethyl-2-propenoate (DCDP). The inhibitor was fed to detached leaves at a concentration of 4 mm, and apparent photosynthesis was measured at atmospheric levels of CO₂ and at 20 and 210 mL L⁻¹ of O₂. Photosynthesis at 210 mL L⁻¹ of O₂ was inhibited 32% by DCDP in F. linearis, by 60% in F. brownii, and by 87% in F. trinervia. Inhibition in the hybrids ranged from 38 to 52%. The inhibition of photosynthesis by 210 mL L⁻¹ of O₂ was increased when DCDP was used, except in the C₄ species, F. trinervia, in which photosynthesis was insensitive to O₂. Except for F. trinervia, control plants with less O₂ sensitivity (more C₄-like) exhibited a progressively greater change in O₂ inhibition of photosynthesis when treated with DCDP. This increased O₂ inhibition probably resulted from decreased CO₂ concentrations in bundle sheath cells due to inhibition of phosphoenolpyruvate carboxylase. The inhibition of photosynthesis by DCDP is concluded to underestimate the degree of C₄ photosynthesis in the interspecific hybrids because increased direct assimilation of atmospheric CO₂ by ribulose bisphosphate carboxylase may compensate for inhibition of phosphoenolpyruvate carboxylase.     

The Influence of Leaf Age on C(4) Photosynthesis and the Accumulation of Inorganic Carbon in Flaveria trinervia, a C(4) Dicot

Characteristics of C₄ photosynthesis were examined in young, mid-age, and mature leaves of Flaveria trinervia (an NADP-malic enzyme-type C₄ dicot). The turnover of [4-¹⁴C] (malate plus aspartate) following a pulse with ¹⁴CO₂ was similar in leaves of different ages (apparent half-time of 18-25 seconds). However, the rate of ¹⁴CO₂ incorporation in mid-age leaves was about 1.5-fold higher than in young leaves, and about 2.5-fold higher than in mature leaves. The rate of ¹⁴CO₂ fixation was proportional to the total active pool of malate plus aspartate but was not correlated with the total photosynthetically derived inorganic carbon pool. The leaf's ability to concentrate inorganic carbon photosynthetically declined during leaf expansion, from 29 down to 7 nanomoles per milligram chlorophyll. Similarly, the active aspartate pool also declined during leaf expansion, from about 123 down to 20 nanomoles per milligram chlorophyll. Enhanced metabolism of aspartate to CO₂ and pyruvate in young leaves is suggested to facilitate the maintenance of high CO₂ levels in bundle sheath cells which are thought to have a higher conductance to CO₂.     

Photosynthesis in Flaveria brownii, a C(4)-Like Species: Leaf Anatomy, Characteristics of CO(2) Exchange, Compartmentation of Photosynthetic Enzymes, and Metabolism of CO(2)

Light microscopic examination of leaf cross-sections showed that Flaveria brownii A. M. Powell exhibits Kranz anatomy, in which distinct, chloroplast-containing bundle sheath cells are surrounded by two types of mesophyll cells. Smaller mesophyll cells containing many chloroplasts are arranged around the bundle sheath cells. Larger, spongy mesophyll cells, having fewer chloroplasts, are located between the smaller mesophyll cells and the epidermis. F. brownii has very low CO₂ compensation points at different O₂ levels, which is typical of C₄ plants, yet it does show about 4% inhibition of net photosynthesis by 21% O₂ at 30°C. Protoplasts of the three photosynthetic leaf cell types were isolated according to relative differences in their buoyant densities. On a chlorophyll basis, the activities of phosphoenolpyruvate carboxylase and pyruvate, Pi dikinase (carboxylation phase of C₄ pathway) were highest in the larger mesophyll protoplasts, intermediate in the smaller mesophyll protoplasts, and lowest, but still present, in the bundle sheath protoplasts. In contrast, activities of ribulose 1,5-bisphosphate carboxylase, other C₃ cycle enzymes, and NADP-malic enzyme showed a reverse gradation, although there were significant activities of these enzymes in mesophyll cells. As indicated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the banding pattern of certain polypeptides of the total soluble proteins from the three cell types also supported the distribution pattern obtained by activity assays of these enzymes. Analysis of initial ¹⁴C products in whole leaves and extrapolation of pulse-labeling curves to zero time indicated that about 80% of the CO₂ is fixed into C₄ acids (malate and aspartate), whereas about 20% of the CO₂ directly enters the C₃ cycle. This is consistent with the high activity of enzymes for CO₂ fixation by the C₄ pathway and the substantial activity of enzymes of the C₃ cycle in the mesophyll cells. Therefore, F. brownii appears to have some capacity for C₃ photosynthesis in the mesophyll cells and should be considered a C₄-like species.     

The Effect of Temperature on the Occurrence of O(2) and CO(2) Insensitive Photosynthesis in Field Grown Plants

The sensitivity of photosynthesis to O₂ and CO₂ was measured in leaves from field grown plants of six species (Phaseolus vulgaris, Capsicum annuum, Lycopersicon esculentum, Scrophularia desertorum, Cardaria draba, and Populus fremontii) from 5°C to 35°C using gas-exchange techniques. In all species but Phaseolus, photosynthesis was insensitive to O₂ in normal air below a species dependent temperature. CO₂ insensitivity occurred under the same conditions that resulted in O₂ insensitivity. A complete loss of O₂ sensitivity occurred up to 22°C in Lycopersicon but only up to 6°C in Scrophularia. In Lycopersicon and Populus, O₂ and CO₂ insensitivity occurred under conditions regularly encountered during the cooler portions of the day. Because O₂ insensitivity is an indicator of feedback limited photosynthesis, these results indicate that feedback limitations can play a role in determining the diurnal carbon gain in the field. At higher partial pressures of CO₂ the temperature at which O₂ insensitivity occurred was higher, indicating that feedback limitations in the field will become more important as the CO₂ concentration in the atmosphere increases.     

Genetic and physiological characterization of Flaveria linearis plants having a reduced activity of cytosolic fructose-1.6-bisphosphatase

We investigated the genetic control of cytosolic fructose-1,6-bisphosphatase (cytFBPase) activity, and the relationships between sucrose synthesis capacity and photosynthesis, growth, flowering and whole-plant carbon partitioning in Flaveria linearis Lag. F₁; F₂, and selfed lines generated from plants with low or high cytFBPase activity were used. CytFBPase activity was controlled by one gene and inherited co-dominantly, giving three classes of activity (low, intermediate and high). Reversed O₂ sensitivity of photosynthesis, which indicates an end-product limitation on photosynthesis, was controlled by one gene and co-segregated with low cytFBPase activity. A low activity of cytFBPase decreased the growth rate. A recessive day-neutral flowering trait in Flaveria linearis did not co-segregate with cytFBPase activity. Plants with low cytFBPase activity had an increased shoot-to-root ratio, and flowering caused an additional shift in carbon partitioning to shoots only in plants with low cytFBPase activity. These data indicate that altering sucrose synthesis can affect photosynthesis and plant growth and development.     

Contribution of Metabolites of Photosynthesis to Postillumination CO(2) Assimilation in Response to Lightflects

In the shade plant Alocasia macrorrhiza grown in low light, photosynthetic CO₂ assimilation during a 5 second lightfleck plus postillumination CO₂ assimilation can allow up to 60% more photosynthesis than that which occurs during 5 seconds of steady state light of the same intensity (RL Chazdon, RW Pearcy 1986 Oecologia. 69: 524-531). Metabolites of photosynthesis were measured to determine if the pool of ribulose 1,5-bisphosphate (RuBP) could account for all of the postillumination CO₂ assimilation following a lightfleck in Alocasia. It was found that the pool of triose-P was much larger than that of RuBP and could account for five times more postillumination CO₂ assimilation than could RuBP. The same trend was seen in the sun plant Phaseolus vulgaris when it was grown in the shade. In contrast, sun-grown Alocasia and Phasiolus did not have a large pool of triose-P relative to RuBP following a lightfleck. In sun plants, carbon may rapidly be converted to RuBP in the light whereas in shade plants there may be a restriction in the path between the triose-P and RuBP pools. It is hypothesized that in shade plants the buildup of triose-P rather than RuBP during the lightfleck prevents inhibition of electron transport which may otherwise occur because of competition for ATP between the two kinases of the photosynthetic carbon reduction cycle. Utilization of the triose-P for postillumination CO₂ fixation would require the capacity for significant postillumination ATP synthesis. The extensive grana stacking and large intrathylakoid space which accompanies the high level of chlorophyll in low-light-grown Alocasia could be an important contributing factor to postillumination ATP formation.     

Photosynthesis in Flaveria brownii A.M. Powell : A C(4)-Like C(3)-C(4) Intermediate

Leaves of Flaveria brownii exhibited slightly higher amounts of oxygen inhibition of photosynthesis than the C₄ species, Flaveria trinervia, but considerably less than the C₃ species, Flaveria cronquistii. The photosynthetic responses to intercellular CO₂, light and leaf temperature were much more C₄-like than C₃-like, although 21% oxygen inhibited the photosynthetic rate, depending on conditions, up to 17% of the photosynthesis rate observed in 2% O₂. The quantum yield for CO₂ uptake in F. brownii was slightly higher than that for the C₄ species F. trinervia in 2% O₂, but not significantly different in 21% O₂. The quantum yield was inhibited 10% in the presence of 21% O₂ in F. brownii, yet no significant inhibition was observed in F. trinervia. An inhibition of 27% was observed for the quantum yield of F. cronquistii in the presence of 21% O₂. The photosynthetic response to very low intercellular CO₂ partial pressures exhibited a unique pattern in F. brownii, with a break in the linear slope observed at intercellular CO₂ partial pressure values between 15 and 20 μbar when analyzed in 21% O₂. No significant break was observed when analyzed in 2% O₂. When taken collectively, the gas-exchange results reported here are consistent with previous biochemical studies that report incomplete intercellular compartmentation of the C₃ and C₄ enzymes in this species, and suggest that F. brownii is an advanced, C₄-like C₃-C₄ intermediate.     

Photosynthesis and Activation of Ribulose Bisphosphate Carboxylase in Wheat Seedlings : Regulation by CO(2) and O(2)

Photosynthetic carbon assimilation in plants is regulated by activity of the ribulose 1,5-bisphosphate (RuBP) carboxylase/oxygenase. Although the carboxylase requires CO₂ to activate the enzyme, changes in CO₂ between 100 and 1,400 microliters per liter did not cause changes in activation of the leaf carboxylase in light. With these CO₂ levels and 21% O₂ or 1% or less O₂, the levels of ribulose bisphosphate were high and not limiting for CO₂ fixation. With high leaf ribulose bisphosphate, the K_act(CO₂) of the carboxylase must be lower than in dark, where RuBP is quite low in leaves. When leaves were illuminated in the absence of CO₂ and O₂, activation of the carboxylase dropped to zero while RuBP levels approached the binding site concentration of the carboxylase, probably by forming the inactive enzyme-RuBP complex.

The mechanism for changing activation of the RuBP carboxylase in the light involves not only Mg²⁺ and pH changes in the chloroplast stroma, but also the effects of binding RuBP to the enzyme. In light when RuBP is greater than the binding site concentration of the carboxylase, Mg²⁺ and pH most likely determine the ratio of inactive enzyme-RuBP to active enzyme-CO₂-Mg²⁺-RuBP forms. Higher irradiances favor more optimal Mg²⁺ and pH, with greater activation of the carboxylase and increased photosynthesis.

     

Responses to elevated CO2 of Flaveria linearis plants having a reduced activity of cytosolic fructose-1,6-bisphosphatase

Wide variation exists in the growth responses of C₃ plants to elevated CO₂ levels. To investigate the role of photosynthetic feedback in this phenomenon, photosynthetic parameters and growth were measured for lines of Flaveria linearis with low, intermediate or high cytosolic fructose-1,6-bisphosphatase (cytFBPase) activity when grown at either 35 or 65 Pa CO₂. The effects of pot size on the responses of these lines to elevated CO₂ were also examined. Photosynthesis and growth of plants with low cytFBPase activity were less responsive to elevated CO₂, and these plants had a reduced maximum potential for photosynthesis and growth. Plants with intermediate cytFBPase activity also showed a lower relative growth enhancement when grown at 65 Pa CO₂. There was a significant pot size effect on photosynthesis and growth for line 85-1 (high cytFBPase). This effect was greatest for line 85-1 when grown at 35 Pa CO₂, since these plants showed the greatest downward acclimation of photosynthesis when grown in small pots. There was a minimal pot size effect for line 84-9 (low cytFBPase), and this could be partly attributed to the reduced CO₂ sensitivity of this line. It is proposed that the capacity for sucrose synthesis in C₃, plants is partly responsible for their wide variation in CO₂ responsiveness.     

Theoretical Considerations when Estimating the Mesophyll Conductance to CO(2) Flux by Analysis of the Response of Photosynthesis to CO(2)

The conductance for CO₂ diffusion in the mesophyll of leaves can limit photosynthesis. We have studied two methods for determining the mesophyll conductance to CO₂ diffusion in leaves. We generated an ideal set of photosynthesis rates over a range of partial pressures of CO₂ in the stroma and studied the effect of altering the mesophyll diffusion conductance on the measured response of photosynthesis to intercellular CO₂ partial pressure. We used the ideal data set to test the sensitivity of the two methods to small errors in the parameters used to determine mesophyll conductance. The two methods were also used to determine mesophyll conductance of several leaves using measured rather than ideal data sets. It is concluded that both methods can be used to determine mesophyll conductance and each method has particular strengths. We believe both methods will prove useful in the future.     

Chromosome evolution in the Gibasis linearis group (Commelinaceae)

Karyotypes, DNA amounts, and meiotic behaviour were examined in population samples of two closely related species, Gibasis venustula and G. heterophylla, and their F₁ hybrids. All samples were diploid (2n=12). DNA amount was similar in G. heterophylla, G. venustula ssp. robusta, and some populations of G. venustula ssp. venustula but in 13 samples of the latter, it showed as much as a 60% difference despite karyotypic uniformity. Although DNA levels were related to the altitude of the habitat, there was some variation within two morphological and ecological races of subspecies venustula. Analysis of hybrid meiosis and pollen fertility demonstrated the cytogenetical discreteness of populations, races, and subspecies. Populations and races were concluded still to be evolving following ecological isolation. The significance of quantitative and qualitative genome changes in evolution and their bearing on the classification of these species is discussed.