Reduced Apparent Photorespiration by the C(3)-C(4) Intermediate Species, Moricandia arvensis and Panicum milioides

The CO₂/O₂ specificity factor of sucrose gradient purified ribulose 1,5-bisphosphate carboxylase/oxygenase from the C₃-C₄ intermediate plants Moricandia arvensis (79 ± 1) and Panicum milioides (89 ± 2) was similar to the respective values of the enzyme from the closely related C₃ species, Moricandia foetida (80 ± 5) and Panicum laxum (86 ± 2). Thus, the kinetic properties of this bifunctional enzyme do not explain the reduced rates of photorespiration exhibited by either of these intermediate species.     

Differential Oxygen Response of Photosynthesis in Soybean and Panicum milioides

The carbon dioxide compensation concentration of Panicum milioides was less than that of soybean over the range of 15 to 35 C. In soybean (Glycine max [L.] Merr. cv. Wayne), the compensation concentration was directly proportional to O₂ concentration. In P. milioides, the compensation concentration was near zero up to 10% O₂ and then increased linearly with higher O₂, although the slope of the response was less than that in soybean. Leaf extracts of P. milioides contained 3-fold higher phosphoenolpyruvate carboxylase activity than soybean leaf extracts. Oxygen inhibition of photosynthesis and carboxy-lation efficiency was less in P. milioides than that observed in soybean. The affinity of P. millioides ribulose-1,5-di-P carboxylase for CO₂ appeared to be slightly greater than that of soybean. The affinity of both enzymes for O₂ was similar. The reduced response of the compensation concentration and photosynthesis to O₂ in P. milioides may be explained by photosynthetic phosphoenolpyruvate carboxylase fixation and by an apparent increased affinity of ribulose-1,5-di-P carboxylase for CO₂.     

Callus induction and plant regeneration in Panicum bisulcatum and Panicum milioides

Surface sterilized seeds and mesocotyls from sterile seedlings from Panicum bisulcatum Thumb., as well as basal parts of leaves and mesocotyls from sterile seedlings, and seeds from Panicum milioides Nees ex. Trin were used as explants to induce callus on a Murashige and Skoog medium supplemented with 2.5 to 10 mg/l of 2,4-D. Subculturing of the white callus from P. milioides and of the brown callus from P. bisulcatum on a medium containing 0.1 mg/l 2,4-D and 10 g/l sucrose led in both species to the appearance of green structures from which plants could be regenerated. Plants were regenerated by an organogenetic process in P. milioides, while P. bisulcatum plants were regenerated both via organogenesis and somatic embryogenesis. 1032 and 94 plants, from P. bisulcatum and P. milioides, respectively, were transferred into soil, and about 90% of them were grown to maturity and set seeds.Abbreviations MS Murashige and Skoog medium (15) - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indoleacetic acid     

Panicum milioides, a Gramineae plant having Kranz leaf anatomy without C4-photosynthesis

Light and electron microscopic observations of the leaf tissueof Panicum milioides showed that the bundle sheath cells containeda substantial number of chloroplasts and other organelles. Theradial arrangement of chlorenchymatous bundle sheath cells,designated as Kranz leaf anatomy, has been considered to bespecific to C₄ plants. However, photosynthetic ¹⁴CO₂ fixationand ¹⁴CO₂ pulse-and-chase experiments revealed that the reductivepentosephosphate pathway was the main route operating in leavesof P. milioides. The interveinal distance of the leaves wasintermediate between C₃and C₄Gramineae species. These resultsindicate that P. milioides is a natural plant species havingchracteristics intermediate between C₃ and C₄ types. (Received March 6, 1975; )     

Phosphoenolpyruvate carboxylase reduces photorespiration in Panicum milioides, a C3-C4 intermediate species.

Panicum milioides represents the first well-documented example of a higher plant species with reduced photorespiration and O₂ inhibition of photosynthesis. We have investigated the biochemical mechanism(s) involved in reducing O₂ sensitivity of photosynthesis in this species by parallel enzyme inhibitor experiments with thin leaf slices of P. milioides and C₃ and C₄Panicum species. The reduced O₂ sensitivity of net photosynthesis in P. milioides gradually increased with increasing concentrations of the phosphoenolpyruvate carboxylase (EC 4.1.1.31) inhibitors, maleate and malonate. At saturating levels of inhibitor, photosynthesis in 2% O₂ was decreased by about 18%, and the inhibitory effects of both 21% O₂ and 49% O₂ were identical to those observed with a C₃Panicum species in the absence or presence of inhibitor. A significant potential for C₄ photosynthesis in P. milioides, compared to its complete absence in a C₃Panicum species, was demonstrated on the basis of: (a) a coupling of leaf slice CO₂ fixation by phosphoenolpyruvate carboxylase with the C₃ cycle; (b) NAD-malic enzyme (EC 1.1.1.39)-dependent aspartate and malate decarboxylation in leaf slices; (c) a full complement of C₄ cycle enzymes in leaf extracts, including pyruvate, P_i dikinase (EC 2.7.9.1) and NAD-malic enzyme; and (d) Kranz-like leaf anatomy with numerous plasmodesmata traversing the mesophyll-bundle sheath interfacial cell wall. These data indicate that the reduced photorespiration and O₂ inhibition of photosynthesis in P. milioides is due to phosphoenolpyruvate carboxylase participation, possibly by creating a limited C₄-like CO₂ pump, rather than an altered ribulose 1,5-bisphosphate carboxylase-oxygenase (EC 4.1.1.39).     

Photosynthesis and Photorespiration in Typha latifolia

Photosynthetic rates of Typha latifolia, the broad-leaved cattail, are the equivalent of rates reported in tropical grasses and other plants which assimilate carbon by the phosphopyruvate carboxylase reaction, but photosynthesis in T. latifolia proceeds by a typical Calvin cycle. Glycolate oxidase, the photorespiratory enzyme, is present in high concentration in this species, but only minor quantities of the assimilated carbon pass through the photorespiratory pathway. However, continued operation of the pathway is apparently essential in the maintenance of assimilatory capacity. Glycolate oxidase function is not closely coupled to stomatal operation in T. latifolia.     

Photosynthetic efficiency of Panicum hians and Panicum milioides in relation to C3 and C4 plants

Panicum hians and Panicum milioides were found to have characteristicsintermediate to those of C₃ and C₄ species with respect to CO₂compensation point, percentage inhibition of photosynthesisby O₂ at various O₂/CO₂ solubility ratios, and water use efficiency.C₄ species have a higher carboxylation efficiency than eitherthe intermediate or C₃ species. During photosynthesis, evenunder 2.5% O₂, C₄ species have a higher affinity for intercellularCO₂ (Km 1.6 µM) apparently due to the initial carboxylationthrough PEP carboxylase. Under low O₂ the intermediate and C₃species had a similar affinity for intercellular CO₂ duringphotosynthesis (Km 5–7 µM) consistent with carboxylationof atmospheric CO₂ through RuDP carboxylase. There were considerablevariation in photosynthesis/unit leaf area at saturating CO₂levels in the species examined which in part is due to differencesin RuDP carboxylase /unit leaf area. The highest rates of photosynthesis/unitleaf area under CO₂-saturating conditions were with the C₃ specieswhich had a correspondingly high level of RuDP carboxylase/unitleaf area. Possibilities for the greater efficiency of P. hiansand P. milioides in comparison to C₃ species in utilizing lowlevels of CO₂ in the presence of atmospheric O₂ are discussed. ¹ This research was supported by the College of Agriculturaland Life Sciences, University of Wisconsin, Madison; and theUniversity of Wisconsin Research Committee with funds from theWisconsin Alumni Research Foundation. (Received June 25, 1977; )     

Photosynthetic/Photorespiratory Carbon Metabolism in the C(3)-C(4) Intermediate Species, Moricandia arvensis and Panicum milioides

The distribution of ¹⁴C in photosynthetic metabolites of two naturally occurring higher plants with reduced photorespiration, Moricandia arvensis and Panicum milioides, in pulse and pulse-chase ¹⁴CO₂ incorporation experiments was similar to that for the C₃ species, M. foetida and Glycine max. After 6 seconds of ¹⁴CO₂ incorporation, only about 6% of the total ¹⁴C fixed was in malate and aspartate in both M. arvensis and P. milioides. The apparent turnover of the C₄ acids was very slow, and malate accumulated during the day in M. arvensis. Thus, C₄ acid metabolism by M. arvensis and P. milioides had no significant role in photosynthetic carbon assimilation under the conditions of our experiments (310 microliters CO₂ per liter, 21% O₂, 1100 or 1900 micromoles photon per square meter per second, 27°C).

After a 36-second chase period in air containing 270 microliters CO₂ per liter, about 20% of the total ¹⁴C fixed was in glycine with M. arvensis, as compared to 15% with M. foetida, 14% with P. milioides, and 9% with G. max. After a 36-second chase period in 100 microliters CO₂ per liter, the percentage in glycine was about twice that at 270 microliters CO₂ per liter in the C₃ species and P. milioides, but only 20% more ¹⁴C was in glycine in M. arvensis. These data suggest that either the photorespiratory glycine pool in M. arvensis is larger than in the other species examined or the apparent turnover rate of glycine and the flow of carbon into glycine during photorespiration are less in M. arvensis. An unusual glycine metabolism in M. arvensis may be linked to the mechanism of photorespiratory reduction in this crucifer.

     

Photosynthesis, Photorespiration and Respiration of Chloroplasts From Acetabularia mediterrania

A chloroplast fraction isolated from Acetabularia mediterrania carries on photosynthesis at rates essentially equal to those of whole cells. Electron and phase contrast microscopy reveals that the chloroplasts are intact and well preserved. Preparations contain no identifiable peroxisomes, but some cytoplasmic and mitochondrial contamination is present. Photosynthesis and CO₂ production in light by chloroplast preparations are in many respects similar to that of bean leaves, although the measured rates are somewhat lower. Respiration and photosynthesis of chloroplast preparations and whole cells of Acetabularia is essentially similar except that cells have a strong dark-type respiration which continues in light and is CO₂ dependent, the substrate being mainly recent photosynthate. The data suggest that chloroplasts are the site of photorespiration.     

Effect of Low Relative Humidity on {delta}13C Value in Two C3 Grasses and in Panicum milioides, a C3-C4 Intermediate Species

When grown under conditions of low relative humidity, the C₃–C₄intermediate Panicum milioides, as well as the C₃ grasses Triticumaestivum and Poa pratense, exhibited ¹³C values which were upto 2–7%o less negative than the ¹³C values of the correspondingplants grown at high relative humidity. At both humidity levels,there was no evidence of a substantial contribution of phosphoenolpyruvatecarboxylase to carbon gain in Panicum milioides     

Photorespiration and Oxygen Inhibition of Photosynthesis in Chlorella pyrenoidosa

The inhibition of photosynthesis by O₂ in air-grown Chlorella pyrenoidosa was investigated using three experimental techniques (artificial leaf, aqueous method, and O₂ electrode) to measure carbon assimilation. CO₂ response curves were determined under different O₂, pH, and temperature conditions. Regardless of the experimental technique and condition, O₂ inhibition was not evident until a concentration of 50% was reached; V_max values were reduced whereas K_m (CO₂) values were unaffected by the increasing O₂ concentration. The response of photosynthesis to O₂ was independent of CO₂ and HCO₃⁻ concentrations as well as temperature. Relative rates of photosynthesis showed a 4 to 5% stimulation in 2% O₂, a 12% inhibition in 50% O₂, and a 24% inhibition in 100% O₂. The inhibition by 50% O₂ was still reversible after 20 minutes exposure whereas 100% O₂ caused irreversible inhibition after only 4 minutes.     

Photosynthesis and Photorespiration in Lolium multiflorum and Lolium perenne

The response of net photosynthesis to changing light-flux densityby leaves of Lolium multiflorum (S. 22) and L. perenne (S. 321)is more adequately described by current models when a term allowingfor photorespiration is included. The magnitude of this termwas determined from the changes in the slope of the light-responsecurves for net photosynthesis. A pseudo first-order rate-constantfor photorespiration, and a pseudo second-order rate-constantfor photosynthesis calculated by this technique for L. multiflorumwere found to be similar to corresponding parameters calculatedfrom light-compensation-point measurements using a simple modeldescribed by Brown (1969). The relative magnitudes of respirationand photosynthesis at light saturation for both Lolium specieswere similar to reported values for other temperate species(Lake, 1967). Two selection lines of L. perenne (S. 321) with contrastingdry-matter yields were found to have the same parameters forrespiration and photosynthesis.     

Effects of Sodium on Photosynthesis in Panicum coloratum

Foliar application of NaCl to sodium-deficient Panicum coloratumstimulated photosynthesis, as did application via roots. Effectsof sodium on photosynthetic responses to internal concentrationsof CO₂ under different light intensities and initial productsof ¹⁴CO₂ fixation suggested that CO₂ fixation and aminationof oxalacetate were limited by sodium deficiency. ² Present address: Institute for Life Science Research, NihonNohyaku Co., Ltd., Kawachi-Nagano, Osaka, 586 Japan.     

A Mutant of Chlamydomonas reinhardtii with Reduced Rate of Photorespiration

Photorespiration rates under air-equilibrated conditions (0.04%CO₂ and 21% O₂) were measured in Chlamydomonas reinhardtii wild-type2137, a phosphoglycolate-phosphatase-deficient (pgp1) mutantand a suppressor double mutant (7FR2N) derived from the pgp1mutant. In both cells grown under 5% CO₂ and adapted air for24 h in the suppressor double mutant, the maximal rate of photorespiration(phosphoglycolate synthesis) was only about half of that ineither the wild type or the pgp1 mutant (18-7F) cells. In theprogeny, the reduced rate of photorespiration was accompaniedby increased photosynthetic affinity for inorganic carbon andthe capacity for growth under air whether accompanied by thepgp1 background or not. Tetrad analyses suggested that thesethree characteristics all resulted from a nuclear single-genemutation at a site unlinked to the pgp1 mutation. The decreasein photorespiration was, however, not due to an increase inthe CO₂/O₂ relative specificity of ribulose-1,5-bisphosphatecarboxylase/oxygenase of 7FR2N or of any other suppressor doublemutants tested. The relationship between the decrease in therate of photorespiration and the CO₂-concentrating mechanismis discussed. ³ Current address: Institute of Botany, Academy of Sciences,Patamdar Shosse, 40, Baku, 370073, Azerbaijan. ⁴ Current address: Department of Management and InformationScience, Jobu University, 270-1, Shinmachi, Tano, Gunma, 370-1393Japan.     

pH Dependence of Photosynthesis and Photorespiration in Soybean Leaf Cells

The effect of pH on the kinetics of photosynthesis, O(2) inhibition of photosynthesis, and photorespiration was examined with mesophyll cells isolated from soybean (Glycine max [L.] Merr.) leaves. At constant, subsaturating bicarbonate concentration (0.5 mm), O(2) inhibition of photosynthesis increased with increasing pH because high pH shifts the CO(2)-bicarbonate equilibrium toward bicarbonate, thereby reducing the CO(2) concentration. At constant, substrating CO(2) concentrations, cell photorespiration decreased with increasing pH. This was indicated by decreases in the CO(2) compensation concentration, O(2) inhibition of photosynthesis, and glycine synthesis. Km(CO(2)) values for isolated cell photosynthesis and in vitro ribulose-1, 5-diphosphate carboxylase activity decreased with increasing pH, while the Ki(O(2)) for both systems was similar at all pH values. The responses to pH of the corresponding kinetic constants of cell photosynthesis and in vitro RuDP carboxylase with respect to CO(2) and O(2) were identical. This provides additional evidence that the relative rates of photosynthesis and photorespiration in C(3) plants are determined by the kinetic properties of RuDP carboxylase.     

Photosynthesis of Grass Species Differing in Carbon Dioxide Fixation Pathways : VIII. Ultrastructural Characteristics of Panicum Species in the Laxa Group

Ultrastructural studies of leaves of seven Panicum species in or closely related to the Laxa group and classified as C₃, C₄ or C₃-C₄ intermediate were undertaken to examine features associated with C₃ and C₄ photosynthesis. The C₃ species Panicum rivulare Trin. had few organelles in bundle sheath cell profiles (2 chloroplasts, 1.1 mitochondria, and 0.3 peroxisomes per cell section) compared to an average of 10.6 chloroplasts, 17.7 mitochondria, and 3.2 peroxisomes per bundle sheath cell profile for three C₃-C₄ species, Panicum milioides Nees ex Trin., Panicum decipiens Nees ex Trin. and Panicum schenckii Hack. However, two other C₃ species, Panicum laxum Sw. and Panicum hylaeicum Mez, contained about 0.7, 0.5, and 0.3 as many chloroplasts, mitochondria, and peroxisomes, respectively, as in bundle sheath cell profiles of the C₃-C₄ species. Chloroplasts and mitochondria in bundle sheath cells were larger than those in mesophyll cells for the C₄ species Panicum prionitis Griseb. and the C₃-C₄ species, but in C₃ species the organelles were similar in size or were smaller in the bundle sheath cells. The C₃-C₄ species and P. laxum and P. hylaeicum exhibited an unusually close association of organelles in bundle sheath cells with mitochondria frequently surrounded in profile by chloroplasts. The high concentrations in bundle sheath cells of somewhat larger organelles than in mesophyll cells correlates with the reduced photorespiration of the C₃-C₄ species.     

不同磷营养水平对烟草叶片光合作用和光呼吸的影响(英文)

研究了不同磷营养水平(0,1/4,1/2,1,2 P)对烟草(Nicotiana rustica L.)叶片光合、光呼吸、乙醇酸合成和乙醇酸氧化酶活性的影响,结果如下; 光合强度在0～1P范围内随磷水平的提高而增高,但在2P水平中略为下降。光呼吸强度在1/4～2P范围内与光合强度有相同的变化趋势,但在磷水平为零时最高;光呼吸/光合比值亦在磷水平为零时最高,并随磷营养的增加而下降。 HPMS抑制乙醇酸氧化酶活性,乙醇酸的积累量随磷水平的变化与光呼吸有一致的趋势。 在加入FMN时,不同磷营养水平的烟草叶片中乙醇酸氧化酶的活性随磷水平的提高而下降;加入FMN对酶活性的促进作用亦随磷水平的提高而下降。 叶片无机磷、有机磷及总磷含量均随磷营养水平的提高而增加。 用不同浓度的磷酸盐溶液真空渗入在1P培养的烟草叶圆片,并在0.25,0.5,10mmol/LNaHCO_3溶液中测定光合和光呼吸,结果表明在0.25和 0.5mmol/L NaHCO_3中,光呼吸随磷浓度的增加而下降;在 10mmol/L中光呼吸完全受抑制。光合作用与磷浓度关系呈单峰曲线,随着NaHCO_3浓度的提高,其高峰位置向右移,即光合最适磷浓度增大。 根据试验结果及从化学计量学推算,认为磷营养有抑制光呼吸的作用,而光呼吸的运转则有补充叶绿体内进行光合作用所需的无机磷的功能。     

Photosynthetic carbon metabolism in Panicum milioides, a C3-C4 intermediate species: evidence for a limited C4 dicarboxylic acid pathway of photosynthesis

Panicum milioides, a naturally occurring species with C4-like Kranz leaf anatomy, is intermediate between C3 and C4 plants with respect to photo-respiration and the associated oxygen inhibition of photosynthesis. This paper presents direct evidence for a limited degree of C4 photosynthesis in this C3-C4 intermediate species based on: (a) the appearance of 24% of the total 14C fixed following 4 s photosynthesis in 14CO2-air by excised leaves in malate and aspartate and the complete transfer of label from the C4 acids to Calvin cycle intermediates within a 15 s chase in 12CO2-air; (b) pyruvate- or alanine-enhanced light-dependent CO2 fixation and pyruvate stimulation ote- or alanine-enhanced light-dependent CO2 fixation and pyruvate stimulation of oxaloacetate- or 3-phosphoglycerate-dependent O2 evolution by illuminated mesophyll protoplasts, but not bundle sheath strands; and (c) NAD-malic enzyme-dependent decarboxylation of C4 acids at the C-4 carboxyl position, C4 acid-dependent O2 evolution, and 14CO2 donation from (4-14C)C4 acids to Calvin cycle intermediates during photosynthesis by bundle sheath strands, but not mesophyll protoplasts. However, P. milloides differs from C4 plants in that the activity of the C4 cycle enzymes is only 15 to 30% of a C4 Panicum species and the Calvin cycle and phosphoenolpyruvate carboxylase are present in both cell types. From these and related studies (Rathnam, C.K.M. and Chollet, R. (1979) Arch. Biochem. Biophys. 193, 346-354; (1978) Biochem. Biophys. Res. Commun. 85, 801-808) we conclude that reduced photorespiration in P. milioides is due to a limited degree of NAD-malic enzyme-type C4 photosynthesis permitting an increase in pCO2 at the site of bundle sheath, but not mesophyll, ribulose-bisphosphate carboxylase-oxygenase.     

不同磷营养水平对烟草叶片光合作用和光呼吸的影响

随着磷营养水平的提高,烟草叶片的CO_2补偿点下降、光合速率上升。光呼吸在缺磷时最高。用光呼吸抑制剂处理烟草叶片后,光合的最适磷浓度提高。当CO_2浓度为560μl/L时,缺磷的烟草叶片在从21％O_2转入2％O_2时出现光合振荡,表明光呼吸与磷营养有密切关系。光呼吸在形成乙醇酸时所释放的磷,有回补叶绿体进行光合作用所需的磷的作用。     

Photosynthesis, Morphology, Leaf Anatomy, and Cytogenetics of Hybrids between C(3) and C(3)/C(4)Panicum Species

The Laxa group of the Panicum genus contains species which have CO₂ exchange and anatomical characteristics intermediate to C₃ and C₄ photosynthetic types (C₃/C₄), and also species characterized as C₃. Hybrids were made between two of the C₃/C₄ species and two C₃ species. Carbon dioxide exchange and morphological, leaf anatomical, and cytogenetic characteristics of F₁ hybrids between Panicum milioides Nees. ex Trin (C₃/C₄) and P. laxum Mez. (C₃), P. spathellosum Doell (C₃/C₄) and P. boliviense Hack. (C₃), and P. spathellosum and P. laxum were studied. There were no consistent differences in apparent photosynthesis, although two of the three hybrids had higher net CO₂ uptake than the C₃ parent. Values of inhibition of apparent photosynthesis by 21% O₂, CO₂ loss in the light, and CO₂ compensation concentration for the hybrids were between those of the parents. All three hybrids showed leaf anatomical traits, especially organelle quantities in the bundle sheath cells, between those of their respective parents. Linear regression of CO₂ compensation concentration on the percentage of mitochondria and chloroplasts in vascular bundle sheaths of the parents and hybrids gave correlation coefficients of −0.94. This suggests that the reduction in CO₂ loss in the C₃/C₄ species, and to a lesser degree in the F₁ hybrids, was due to development of organelles and perhaps a higher proportion of leaf photorespiration in bundle sheaths. The overall morphology of the hybrids was so different from the parents that they could be described as new taxonomic forms. The chromosomes in the hybrids were mainly unpaired or paired as bivalents indicating possible homology between some parental genomes.