ABSTRACT
The succulent, cylindrical leaves of the C(4) dicot Portulaca grandiflora possess three distinct green cell types: bundle sheath cells (BSC) in radial arrangement around the vascular bundles; mesophyll cells (MC) in an outer layer adjacent to the BSC; and water storage cells (WSC) in the leaf center. Unlike typical Kranz leaf anatomy, the MC do not surround the bundle sheath tissue but occur only in the area between the bundle sheath and the epidermis. Intercellular localization of photosynthetic enzymes was characterized using protoplasts isolated enzymatically from all three green cell types.Like other C(4) plants, P. grandiflora has ribulose 1,5-bisphosphate carboxylase and the decarboxylating enzyme, NADP(+)-malic enzyme, in the BSC. Unlike other C(4) plants, however, phosphoenolpyruvate carboxylase, pyruvate, Pi dikinase, and NADP(+)-malate dehydrogenase of the C(4) pathway were present in all three green cell types, indicating that all are capable of fixing CO(2) via phosphoenolpyruvate carboxylase and regenerating phosphoenolpyruvate. Other enzymes were about equally distributed between MC and BSC similar to other C(4) plants. The enzyme profile of the WSC was similar to that of the MC but with reduced activity in most enzymes, except mitochondrion-associated enzymes.Intracellular localization of enzymes was studied in organelles partitioned by differential centrifugation using mechanically ruptured mesophyll and bundle sheath protoplasts. Phosphoenolpyruvate carboxylase was a cytosolic enzyme in both cells; whereas, ribulose 1,5-bisphosphate carboxylase and NADP(+)-malic enzyme were exclusively compartmentalized in the bundle sheath chloroplasts. NADP(+)-malate dehydrogenase, pyruvate, Pi dikinase, aspartate aminotransferase, 3-phosphoglycerate kinase, and NADP(+)-triose-P dehydrogenase were predominantly localized in the chloroplasts while alanine aminotransferase and NAD(+)-malate dehydrogenase were mainly present in the cytosol of both cell types. Based on enzyme localization, a scheme of C(4) photosynthesis in P. grandiflora is proposed.Well-watered plants of P. grandiflora exhibit a diurnal fluctuation of total titratable acidity, with an amplitude of 61 and 54 microequivalent per gram fresh weight for the leaves and stems, respectively. These changes were in parallel with changes in malic acid concentration in these tissues. Under severe drought conditions, diurnal changes in both titratable acidity and malic acid concentration in both leaves and stems were much reduced. However, another C(4) dicot Amaranthus graecizans (nonsucculent) did not show any diurnal acid fluctuation under the same conditions. These results confirm the suggestion made by Koch and Kennedy (Plant Physiol. 65: 193-197, 1980) that succulent C(4) dicots can exhibit an acid metabolism similar to Crassulacean acid metabolism plants in certain environments.
ABSTRACT
In leaves of plants with C(4) photosynthesis, sulfur assimilation is initiated in bundle sheath cells whereas carbon and nitrogen assimilation are initiated in mesophyll cells. The activation of sulfate by adenosine triphosphate sulfurylase in leaves of C(4) plants occurs in chloroplasts of bundle sheath cells and is effected by two isozymes of approximately equal activities that accounted for 95 to 100 percent of the total leaf activity.
ABSTRACT
In the C4 plant, Amaranthus graecizans, increasing [O2] from 2% up to 100% inhibited photosynthesis, quantum yield, and the carboxylation efficiency, and increased the CO2 compensation point (Γ) from 2 to about 12 µl/l. The O2 inhibition of photosynthesis was fully reversible. When changing from 2.5 to 40% O2 and vice versa, about 1 h was required for full equilibration with an O2 inhibition of 18%; whereas in wheat, a C3 species, inhibition of photosynthesis and its reversal occurs within minutes after changing [O2], resulting in 63% inhibition of photosynthesis by 45% O2. These differences in O2 inhibition between a C4 and C3 species can be explained by high diffusive resistance across bundle-sheath cells of C4 plants and the increased CO2/O2 ratio in bundle-sheath cells which is the consequence of the C4 cycle. In A. graecizans, Γ increased with increasing [O2] but tended to reach a maximum at relatively high O2 levels. The lack of a linear increase in Γ as previously observed for C3 species indicates that a considerable amount of photorespired CO2 may be re-fixed with increasing levels of O2. In comparison to previous reports with other C4 species, photosynthesis of A. graecizans shows greater sensitivity to O2, with a noticeable inhibition occurring with shifts from 2 to 21% O2. A. graecizans has characteristics of other C4 species with respect to Kranz anatomy, localization of PEP carboxylase in mesophyll cells and RuBP carboxylase in bundle-sheath cells, and little fractionation among carbon isotopes during CO2 fixation. The basis for the higher sensitivity of photosynthesis of A. graecizans to O2 may be based upon a lower diffusive resistance of gases across bundle-sheath cells than in some other C4 species.
ABSTRACT
The intracellular locations of six key enzymes of Crassulacean acid metabolism were determined using enzymically isolated mesophyll protoplasts of Sedum praealtum D.C. Data from isopycnic sucrose density gradient centrifugation established the chloroplastic location of pyruvate Pi dikinase, the mitochondrial location of NAD-linked malic enzyme, and exclusively nonparticulate (not associated with chloroplasts, peroxisomes, or mitochondria) locations of phosphoenolpyruvate carboxylase, NADP-linked malic enzyme, enolase, and phosphoglycerate mutase. The consequences of this enzyme distribution with respect to compartmentalization of the pathway and the transport of metabolites in Crassulacean acid metabolism are discussed.
ABSTRACT
The effect of leaf temperature, O2 and calculated O2/CO2 solubility ratio in the leaf on the quantum yield of photosynthesis was studied for the C4 species, Zea mays L., and the C3 species, Triticum aestivum L. Over a range of leaf temperatures of 16 to 35° C, the quantum yield of Z. mays was relatively constant and was similar under 1.5 and 21% O2, being ca. 0.059 mol CO2 mol(-1) quanta absorbed. Under 1.5% O2 and atmospheric levels of CO2, the quantum yield of T. aestivum was relatively constant (0.083 mol CO2 mol(-1) quanta absorbed) at leaf temperatures from 15 to 35° C. Atmospheric levels of O2 (21%) reduced the quantum yield of photosynthesis in T. aestivum and as leaf temperature increased, the quantum yield decreased from 0.062 at 15°C to 0.046 mol CO2 mol(-1) quanta absorbed at 35°C. Increasing temperature decreases the solubility of CO2 relatively more than the solubility of O2, resulting in an increased solubility ratio of O2/CO2. Experimentally manipulating the atmospheric levels of O2 or CO2 to maintain a near-constant solubility ratio of O2/CO2 at varying leaf temperatures largely prevented the temperature-dependent decrease in quantum yield in t. aestivum. Thus, the decrease in quantum yield with increasing leaf temperature in C3 species may be largely caused by a temperaturedependent change in the solubility ratio of O2/CO2.
ABSTRACT
Individual leaves of potato (Solanum tuberosum L. W729R), a C(3) plant, were subjected to various irradiances (400-700 nm), CO(2) levels, and temperatures in a controlled-environment chamber. As irradiance increased, stomatal and mesophyll resistance exerted a strong and some-what paralleled regulation of photosynthesis as both showed a similar decrease reaching a minimum at about 85 neinsteins.cm(-2).sec(-1) (about (1/2) of full sunlight). Also, there was a proportional hyperbolic increase in transpiration and photosynthesis with increasing irradiance up to 85 neinsteins.cm(-2).sec(-1). These results contrast with many C(3) plants that have a near full opening of stomata at much less light than is required for saturation of photosynthesis.Inhibition of photosynthesis by 21% O(2) was nearly overcome by a 2-fold increase in atmospheric levels of CO(2) (about 1,200 ng.cm(-3)). Photosynthesis at 25 C, high irradiance, 2.5% O(2) and atmospheric levels of CO(2) was about 80% of the CO(2)-saturated rate, suggesting that CO(2) can be rate-limiting even without O(2) inhibition of photosynthesis. With increasing CO(2) concentration, mesophyll resistance decreased slightly while stomatal resistance increased markedly above 550 ng.cm(-3) which resulted in a significant reduction in transpiration.Although potato is a very productive C(3) crop, there is substantial O(2) inhibition of photosynthesis. The level of O(2) inhibition was maximum around 25 C but the percentage inhibition of photosynthesis by O(2) increased steadily from 38% at 16 C to 56% at 36 C. Photosynthesis and transpiration showed broad temperature optima (16-25 C). At higher temperatures, both the increased percentage inhibition of photosynthesis by O(2) and the increased stomatal resistance limit photosynthesis, while increased stomatal resistance limits transpiration. Water use efficiency, when considered at a constant vapor pressure gradient, increased with increasing irradiance, CO(2) concentration, and temperature.
ABSTRACT
The magnitude of the percentage inhibition of photosynthesis by atmospheric levels of O(2) in the C(3) species Solanum tuberosum L., Medicago sativa L., Phaseolus vulgaris L., Glycine max L., and Triticum aestivum L. increases in a similar manner with an increase in the apparent solubility ratio of O(2)/CO(2) in the leaf over a range of solubility ratios from 25 to 45. The solubility ratio is based on calculated levels of O(2) and CO(2) in the intercellular spaces of leaves as derived from whole leaf measurements of photosynthesis and transpiration. The solubility ratio of O(2)/CO(2) can be increased by increased leaf temperature under constant atmospheric levels of O(2) and CO(2) (since O(2) is relatively more soluble than CO(2) with increasing temperature); by increasing the relative levels of O(2)/CO(2) in the atmosphere at a given leaf temperature, or by increased stomatal resistance. If the solubility ratio of O(2)/CO(2) is kept constant, as leaf temperature is increased, by varying the levels of O(2) or CO(2) in the atmosphere, then the percentage inhibition of photosynthesis by O(2) is similar. The decreased solubility of CO(2) relative to O(2) (decreased CO(2)/O(2) ratio) may be partly responsible for the increased percentage inhibition of photosynthesis by O(2) under atmospheric conditions with increasing temperature.
ABSTRACT
The response of whole leaf photosynthesis of wheat (Triticum aestivum L.) in relation to soluble CO(2) available to the mesophyll cells, under low (1.5%) O(2) at 25, 30, and 35 C, followed Michaelis-Menten kinetics up to saturating CO(2) but deviated at high CO(2) levels where the experimental V(max) is considerably less than the calculated V(max). The affinity of the leaves for CO(2) during photosynthesis was similar from 25 to 35 C with Km (CO(2)) values of approximately 3.5 to 5 muM.In considering the effect of O(2) on photosynthesis at 25, 30, and 35 C where O(2) and CO(2) are expressed on a solubility basis: (a) the effect of O(2) on carboxylation efficiency was similar at the three temperature; (b) increasing temperature caused only a slight increase in kinetic constants Ki(O(2)) and Km(CO(2)), while the ratio of Ki(O(2))/Km(CO(2)) was similar at the three temperatures; and (c) the reciprocal plots of apparent rate of photosynthesis versus (CO(2) - Gamma) at various O(2) levels showed O(2) to be a competitive inhibitor of photosynthesis.A model for separating O(2) inhibition of photosynthesis into two components, direct competitive inhibition and inhibition due to photorespiration, was presented from both simulated and experimental data of photosynthetic response curves to varying CO(2) concentrations at low O(2)versus 21% O(2). The photorespiratory part of O(2) inhibition is considered as a major component at Gamma and increases with increasing temperature and with increase in O(2)/CO(2) solubility ratio. The competitive component of O(2) inhibition is considered as a major component of O(2) inhibition under atmospheric CO(2) levels and is relatively independent of temperature at a given O(2)/CO(2) ratio.
ABSTRACT
Mesophyll protoplasts and bundle-sheath cells of Pennisetum purpureum Schum., a C4 plant with low phenol-oxidase activity, were enzymatically separated according to methods recently developed with sugarcane (Saccharum officinarum L.), maize (Zea mays L.), and sorghum (Sorghum bicolor L.). The phosphoenolpyruvate carboxylase and NADP-malic dehydrogenase of the C4 pathway were found to be localized in the mesophyll protoplasts while ribulose-1,5-diphosphate (RuDP) carboxylase, phosphoribulokinase and NADP-malic enzyme were localized in the bundle-sheath cells. The levels of these enzyme activities in the leaf extracts and in certain cellular preparations of P. purpureum are sufficient to account for the rate of photosynthesis in the leaf. These results on the activities and distribution of photosynthetic enzymes with P. purpureum preparations are consistent with our previous evidence for cellular separation of the C4 and the reductive pentose-phosphate pathways in C4 species.With chlorogenic acid as the substrate, P. purpureum, Setaria lutescens (Weigel) Hubb. and Panicum texanum Buckl. have relatively low phenol-oxidase activity, similar to that found in spinach (Spinacia oleracea L.); while sorghum, sugarcane, maize, Panicum capillare L. and P. miliaceum L. have relatively high phenoloxidase activity, similar to that in tobacco (Nicotiana tabacum L.). C4 species having high phenol-oxidase activity have substantial activity of the enzyme in both mesophyll and bundle-sheath extracts. Since phenol oxidase is found in both cell types it is not logical to expect preferential inhibition of RuDP carboxylase or other photosynthetic enzymes through phenol oxidation in mesophyll extracts, as has been previously suggested. When dithiothreitol and polyvinylpyrrolidone were included in the enzyme extraction medium, the activity of RuDP carboxylase increased 10% in P. purpureum and 59% in sugarcane leaf extracts.
