Ammonia emission from rice leaves in relation to photorespiration and genotypic differences in glutamine synthetase activity

Background and Aims

Rice (Oryza sativa) plants lose significant amounts of volatile NH₃ from their leaves, but it has not been shown that this is a consequence of photorespiration. Involvement of photorespiration in NH₃ emission and the role of glutamine synthetase (GS) on NH₃ recycling were investigated using two rice cultivars with different GS activities.

Methods

NH₃ emission (AER), and gross photosynthesis (P_G), transpiration (Tr) and stomatal conductance (g_S) were measured on leaves of ‘Akenohoshi’, a cultivar with high GS activity, and ‘Kasalath’, a cultivar with low GS activity, under different light intensities (200, 500 and 1000 µmol m⁻² s⁻¹), leaf temperatures (27·5, 32·5 and 37·5 °C) and atmospheric O₂ concentrations ([O₂]: 2, 21 and 40 %, corresponding to 20, 210 and 400 mmol mol⁻¹).

Key Results

An increase in [O₂] increased AER in the two cultivars, accompanied by a decrease in P_G due to enhanced photorespiration, but did not greatly influence Tr and g_S. There were significant positive correlations between AER and photorespiration in both cultivars. Increasing light intensity increased AER, P_G, Tr and g_S in both cultivars, whereas increasing leaf temperature increased AER and Tr but slightly decreased P_G and g_S. ‘Kasalath’ (low GS activity) showed higher AER than ‘Akenohoshi’ (high GS activity) at high light intensity, leaf temperature and [O₂].

Conclusions

Our results demonstrate that photorespiration is strongly involved in NH₃ emission by rice leaves and suggest that differences in AER between cultivars result from their different GS activities, which would result in different capacities for reassimilation of photorespiratory NH₃. The results also suggest that NH₃ emission in rice leaves is not directly controlled by transpiration and stomatal conductance.     

Ozone-induced changes in photosynthesis and photorespiration of hybrid poplar in relation to the developmental stage of the leaves

Young poplar trees (Populus tremula Michx. x Populus alba L. clone INRA 717-1B4) were subjected to 120 ppb of ozone for 35 days in phytotronic chambers. Treated trees displayed precocious leaf senescence and visible symptoms of injury (dark brown/black upper surface stippling) exclusively observed on fully expanded leaves. In these leaves, ozone reduced parameters related to photochemistry (Chl content and maximum rate of photosynthetic electron transport) and photosynthetic CO(2) fixation [net CO(2) assimilation, Rubisco (ribulose-1,5-bisphosphate carboxylase oxygenase) activity and maximum velocity of Rubisco for carboxylation]. In fully expanded leaves, the rate of photorespiration as estimated from Chl fluorescence was markedly impaired by the ozone treatment together with the activity of photorespiratory enzymes (Rubisco and glycolate oxidase). Immunoblot analysis revealed a decrease in the content of serine hydroxymethyltransferase in treated mature leaves, while the content of the H subunit of the glycine decarboxylase complex was not modified. Leaves in the early period of expansion were exempt from visible symptoms of injury and remained unaffected as regards all measured parameters. Leaves reaching full expansion under ozone exposure showed potential responses of protection (stimulation of mitochondrial respiration and transitory stomatal closure). Our data underline the major role of leaf phenology in ozone sensitivity of photosynthetic processes and reveal a marked ozone-induced inhibition of photorespiration.     

Isoprene synthase activity and its relation to isoprene emission in Quercus robur L. leaves

Pedunculate oak ( Quercus robur L.) is known as a strong isoprene (2-methyl-1,3-butadiene) emitter. Diurnal changes in isoprene emission were determined by branch enclosure measurements. In contrast to the diurnal cycle in emission rates, specific isoprene synthase activity in the leaves remained unchanged. Based on in vitro enzyme activity and its temperature dependency, an isoprene synthesis capacity at specific leaf temperatures was calculated. The comparison of these 'leaf temperature-dependent enzyme capacities' and the measured emission rates revealed that the enzyme activity of isoprene synthase is comparable to the observed isoprene emission rates. In addition, variation in the isoprene synthase activity of the leaves due to changes in light intensity during leaf development was investigated. A 50% reduction of light intensity by shading of single branches reduced isoprene synthase activity by ≈ 60% compared with full sunlight. The calculation of isoprene synthesis capacities based on enzymatic data obtained under optimum reaction conditions, corrected for actual leaf temperature and related to leaf surface area, provides a sound basis for predicting the isoprene emission potential of plants.     

Effect of the age of tobacco leaves on photosynthesis and photorespiration

Relationships among the activities of enzymes related to photosynthesisand photorespiration, and ¹⁴CO₂ photosynthetic products wereinvestigated with individual tobacco leaves attached to thestalk from the bottom to the top. P-glycolate phosphatase ofthe chloroplasts and glycolate oxidase of the peroxisomes hadtheir maximum activities in the 25th leaf from the dicotyledons.Maximum photorespiration was similarly distributed. The highestratio of serine-¹⁴C to glycine-¹⁴C in the photosynthesates andmaximum glycolate formation were also observed in the 25th leaf.Glutamateglyoxylate aminotransferase, serine hydroxymethyltransferaseand glycine decarboxylase were more active in the upper leaves.RuDP carboxylase had nearly constant activity in all leaves,except for the youngest in which activity decreased. MaximumCO₂ photosynthesis and enzyme activity for the C₄ dicarboxylicacid cycle occurred in the upper, youngest leaf. Distributionof photosynthetic CO² fixation among the leaves did not coincidewith RuDP carboxylase activity. The photosynthetic capacityappeared to be better related to the distribution pattern forenzymes of the C₄ dicarboxylic acid pathway, i.e. PEP carboxylase,pyruvate Pi dikinase and 3-PGA phosphatase in the upper leaves.The results suggest that the C₄ dicarboxylic acid pathway participates,to some extent, in photosynthesis in young leaves of tobacco,a dicotyledonous plant. ¹This work was reported at the Annual Meeting (1970) of theJapanese Plant Physiologists in Kobe. ²The Central Research Institute, Japan Monopoly Corporation1-28-3, Nishishinagawa, Shinagawaku, Tokyo, 141 Japan. (Received November 2, 1972; )     

The influence of light environment on photosynthesis and basal methylbutenol emission from Pinus ponderosa

Methylbutenol is a 5-carbon alcohol that is produced and emitted by several species of pine in western North America, and may have important impacts on the tropospheric chemistry of this region. In the present study the response of methylbutenol basal emission rate (measured at a constant light intensity of 1500 µmol m⁻² s⁻¹ and temperature of 30 °C) to the light and temperature conditions of the growth environment was examined, using field-grown plants shielded with shade cloth of various densities. Methylbutenol basal emission rates increased linearly with the temperature of the growth environment but did not respond to the shading of foliage during growth and development. Both photosynthesis and basal methylbutenol emission rate declined in older needles; however, these declines appear to result from parallel but independent processes and not from basal MBO emission rate directly tracking photosynthetic rates. Older needles did not occupy cooler microenvironments within the canopy; and thus differing thermal microenvironment could not explain the reduced MBO emission in older needles.     

Isoprene emission from plants: why and how

BACKGROUND: Some, but not all, plants emit isoprene. Emission of the related monoterpenes is more universal among plants, but the amount of isoprene emitted from plants dominates the biosphere-atmosphere hydrocarbon exchange. SCOPE: The emission of isoprene from plants affects atmospheric chemistry. Isoprene reacts very rapidly with hydroxyl radicals in the atmosphere making hydroperoxides that can enhance ozone formation. Aerosol formation in the atmosphere may also be influenced by biogenic isoprene. Plants that emit isoprene are better able to tolerate sunlight-induced rapid heating of leaves (heat flecks). They also tolerate ozone and other reactive oxygen species better than non-emitting plants. Expression of the isoprene synthase gene can account for control of isoprene emission capacity as leaves expand. The emission capacity of fully expanded leaves varies through the season but the biochemical control of capacity of mature leaves appears to be at several different points in isoprene metabolism. CONCLUSIONS: The capacity for isoprene emission evolved many times in plants, probably as a mechanism for coping with heat flecks. It also confers tolerance of reactive oxygen species. It is an example of isoprenoids enhancing membrane function, although the mechanism is likely to be different from that of sterols. Understanding the regulation of isoprene emission is advancing rapidly now that the pathway that provides the substrate is known.     

Environmental and developmental controls over the seasonal pattern of isoprene emission from aspen leaves

Isoprene emission from plants represents one of the principal biospheric controls over the oxidative capacity of the continental troposphere. In the study reported here, the seasonal pattern of isoprene emission, and its underlying determinants, were studied for aspen trees growing in the Rocky Mountains of Colorado. The springtime onset of isoprene emission was delayed for up to 4 weeks following leaf emergence, despite the presence of positive net photosynthesis rates. Maximum isoprene emission rates were reached approximately 6 weeks following leaf emergence. During this initial developmental phase, isoprene emission rates were negatively correlated with leaf nitrogen concentrations. During the autumnal decline in isoprene emission, rates were positively correlated with leaf nitrogen concentration. Given past studies that demonstrate a correlation between leaf nitrogen concentration and isoprene emission rate, we conclude that factors other than the amount of leaf nitrogen determine the early-season initiation of isoprene emission. The late-season decline in isoprene emission rate is interpreted as due to the autumnal breakdown of metabolic machinery and loss of leaf nitrogen. In potted aspen trees, leaves that emerged in February and developed under cool, springtime temperatures did not emit isoprene until 23 days after leaf emergence. Leaves that emrged in July and developed in hot, midsummer temperatures emitted isoprene within 6 days. Leaves that had emerged during the cool spring, and had grown for several weeks without emitting isoprene, could be induced to emit isoprene within 2 h of exposure to 32°C. Continued exposure to warm temperatures resulted in a progressive increase in the isoprene emission rate. Thus, temperature appears to be an important determinant of the early season induction of isoprene emission. The seasonal pattern of isoprene emission was examined in trees growing along an elevational gradient in the Colorado Front Range (1829–2896 m). Trees at different elevations exhibited staggered patterns of bud-break and initiation of photosynthesis and isoprene emission in concert with the staggered onset of warm, springtime temperatures. The springtime induction of isoprene emission could be predicted at each of the three sites as the time after bud break required for cumulative temperatures above 0°C to reach approximately 400 degree days. Seasonal temperature acclimation of isoprene emission rate and photosynthesis rate was not observed. The temperature dependence of isoprene emission rate between 20 and 35°C could be accurately predicted during spring and summer using a single algorithm that describes the Arrhenius relationship of enzyme activity. From these results, it is concluded that the early season pattern of isoprene emission is controlled by prevailing temperature and its interaction with developmental processes. The late-season pattern is determined by controls over leaf nitrogen concentration, especially the depletion of leaf nitrogen during senescence. Following early-season induction, isoprene emission rates correlate with photosynthesis rates. During the season there is little acclimation to temperature, so that seasonal modeling simplifies to a single temperature-response algorithm.     

Acclimation of isoprene emission and photosynthesis to growth temperature in hybrid aspen: resolving structural and physiological controls

Acclimation of foliage to growth temperature involves both structural and physiological modifications, but the relative importance of these two mechanisms of acclimation is poorly known, especially for isoprene emission responses. We grew hybrid aspen (Populus tremula x P. tremuloides) under control (day/night temperature of 25/20 °C) and high temperature conditions (35/27 °C) to gain insight into the structural and physiological acclimation controls. Growth at high temperature resulted in larger and thinner leaves with smaller and more densely packed chloroplasts and with lower leaf dry mass per area (M_A). High growth temperature also led to lower photosynthetic and respiration rates, isoprene emission rate and leaf pigment content and isoprene substrate dimethylallyl diphosphate pool size per unit area, but to greater stomatal conductance. However, all physiological characteristics were similar when expressed per unit dry mass, indicating that the area‐based differences were primarily driven by M_A. Acclimation to high temperature further increased heat stability of photosynthesis and increased activation energies for isoprene emission and isoprene synthase rate constant. This study demonstrates that temperature acclimation of photosynthetic and isoprene emission characteristics per unit leaf area were primarily driven by structural modifications, and we argue that future studies investigating acclimation to growth temperature must consider structural modifications.     

Alternate pathways of glycolate synthesis in tobacco and maize leaves in relation to rates of photorespiration

After a preliminary period in light, leaf disks floated on 10 mm α-hydroxy-2-pyridinemethanesulfonic acid to inhibit glycolate oxidase accumulate glycolate at average initial rates of 67 micromoles in tobacco and 8 micromoles per gram fresh weight per hour in maize under optimal conditions in air. In the presence of ¹⁴CO₂, the glycolate synthesized has a high specific radioactivity in illuminated tobacco and a low one in maize. Isonicotinic acid hydrazide also inhibits glycolate oxidation and causes a slow accumulation of glycolate in maize but not in tobacco, while it inhibits glycolate synthesis in tobacco but not in maize. Radioactive carbon in acetate-2-¹⁴C and especially pyruvate-3-¹⁴C is incorporated predominantly into the C-2 of glycolate in both species, but the specific radioactivity is much greater in maize. Glyoxylate-2-¹⁴C is readily converted to glycolate-2-¹⁴C in both species. The addition of phosphoenolpyruvate stimulated glycolate formation in maize and inhibited its synthesis in tobacco, and in the presence of ¹⁴CO₂ the specific radioactivity in glycolate-¹⁴C was decreased greatly by the added phosphoenolpyruvate only in maize.     

Effect of oxygen on photosynthesis, photorespiration and respiration in detached leaves. I. Soybean

The effect of O₂ on the CO₂ exchange of detached soybean leaves was measured with a Clark oxygen electrode and infrared carbon dioxide analysers in both open and closed systems.

The rate of apparent photosynthesis was inhibited by O₂ while the steady rate of respiration after a few minutes in the dark was not affected. Part of the inhibition of apparent photosynthesis was shown to be a result of increased photorespiration. This stimulation of photorespiration by O₂ was manifested by an increase in the CO₂ compensation point.

The differential effects of O₂ on dark respiration (no effect) and photorespiration (stimulation) indicated that these were 2 different processes.

Moreover the extrapolation of the CO₂ compensation point to zero at zero O₂ indicated that dark respiration was suppressed in the light at least at zero O₂ concentration.

     

High carbon dioxide and sun/shade effects on isoprene emission from oak and aspen tree leaves

Abstract. Isoprene (2-methyl 1, 3-butadiene) is emitted from many plants, especially trees. We tested the effect of growth at high CO₂ partial pressure and sun versus shade conditions on the capacity of Quercus rubra L. (red oak) and Populus tremuloides Michx. (quaking aspen) leaves to make isoprene. Oak leaves grown at high CO₂ partial pressure (65 Pa) had twice the rate of isoprene emission as leaves grown at 40Pa CO₂. However, aspen leaves behaved oppositely, with high CO₂-grown leaves having just 60-70% the rate of isoprene emission as leaves grown in 40 Pa CO₂. Similar responses were observed from 25 to 35 °C leaf temperature during assay. The stimulation of isoprene emission by growth at high CO₂ and the stimulation in high temperature resulted in isoprene emission consuming over 15% of the carbon fixed during photosynthesis in high-CO₂ grown oak leaves assayed at 35 °C. Leaves from the south (sunny) sides of trees growing in natural conditions had rates of isoprene emission double those of leaves growing in shaded locations on the same trees. This effect was similar in both aspen and oak. The leaves used for these experiments had significantly different chlorophyll a/b ratios indicating they were functionally sun (from the sunny locations) or shade leaves (from the protected locations). Because the metabolic pathway of isoprene synthesis is unknown, we are unable to speculate about how or why these effects occur. However, these effects are more consistent with metabolic control of isoprene release rather than a metabolic leak of isoprene from metabolism. The results are also important for large scale modelling of isoprene emission and for predicting the effect of future increases in atmospheric CO₂ level on isoprene emission from vegetation.     

Isoprene emissions and photosynthesis in three ferns – The influence of light and temperature

A study was designed to determine the rates of isoprene emission and photosynthesis in three fern species [ Dicksonia antarctica Labill., Thelypteris decursive-pinnata (Van Hall) Ching and Thelypteris kunthii (Desv.) Morton] and the independent influence of light and temperature on these processes. The plants were conditioned in a growth chamber and then transferred to a controlled environment gas-exchange chamber. Samples of the chamber atmosphere were collected; isoprene was concentrated cryo-genically and measured by gas chromatography. Only small amounts of isoprene were detected around the ferns in the dark. Isoprene emissions increased with increasing levels of photosynthetic photon flux density (PPFD) in all three species; 50% of the maximum emission occurred at PPFD levels of 130 to 500 μmol m−² s−¹. Maximum isoprene emissions occurred between 35 and 39°C which is a lower temperature maximum than reported for angiosperms and gymnosperms. The increased emissions with temperature were primarily associated with increased biosynthetic rates for isoprene. Carbon lost through isoprene accounted for 0.02 to 2.6% of the carbon fixed during photosynthesis, depending on the PPFD level, temperature and fern species.     

Carbon dioxide compensation points of leaves and stems and their relation to net photosynthesis

The interactions between CO₂ and H₂O vapor exchange of the leaf and respirant organs like stems were studied in tobacco plants. The results were analyzed according to a suggested model. Good agreement between open and closed system measurements supported the validity of the model.     

Isoprene emission rates under elevated CO2 and O3 in two field-grown aspen clones differing in their sensitivity to O3

Isoprene is the most important nonmethane hydrocarbon emitted by plants. The role of isoprene in the plant is not entirely understood but there is evidence that it might have a protective role against different oxidative stresses originating from heat shock and/or exposure to ozone (O(3)). Thus, plants under stress conditions might benefit by constitutively high or by higher stress-induced isoprene emission rates. In this study, measurements are presented of isoprene emission from aspen (Populus tremuloides) trees grown in the field for several years under elevated CO(2) and O(3). Two aspen clones were investigated: the O(3)-tolerant 271 and the O(3)-sensitive 42E. Isoprene emission decreased significantly both under elevated CO(2) and under elevated O(3) in the O(3)-sensitive clone, but only slightly in the O(3)-tolerant clone. This study demonstrates that long-term-adapted plants are not able to respond to O(3) stress by increasing their isoprene emission rates. However, O(3)-tolerant clones have the capacity to maintain higher amounts of isoprene emission. It is suggested that tolerance to O(3) is explained by a combination of different factors; while the reduction of O(3) uptake is likely to be the most important, the capacity to maintain higher amounts of isoprene is an important factor in strengthening this character.     

Intracellular localization of enzymes related to photorespiration in green leaves

Localization in green leaves of glycine decarboxylase and serinehydroxymethyltransferase (EC 2.1.2.1 [EC] ) was investigated. Subcellularpreparations of green leaves were fractionated by non-linearsucrose isopicnic centrifugation of 2.5 to 1.3 M at 74,700 xor 1.8 to 0.6M at 11,800xg and by centrifugation in non-aqueousmedia. Glycine decarboxylase was located in mitochondria andserine hydroxymethyltransferase was principally located in mitochondriaand partly in chloroplasts. Chloroplastic serine hydroxymethyltransferaseis thought to be responsible for glycine formation from serinederived from photosynthesized 3-phosphoglycerate. A scheme forglycolate metabolism and photorespiration is presented. ¹ A part of this work was presented at the Annual Meeding (April,1969) of the Japanese Society of Plant Physiologists, Kanazawa. ² Department of Biochemistry, Michigan State University, EastLansing, Michigan 48823, U.S.A. (Received August 3, 1970; )     

Benzylaminopurine induced changes in photosynthesis and photorespiration of barley plants

Benzylaminopurine (BA) caused an enhancement of chlorophyll and protein content and a reduced elongation of primary barley leaves. BA did not change the rhythmic pattern of¹⁴CO₂ fixation and activities of RuBP carboxylase, RuBP oxygenase, glycolate oxidase and phosphoglycolate phosphatase, but the enzyme activities were enhanced and the level of¹⁴CO₂ fixation was reduced. Light/dark¹⁴CO₂ evolution ratio was affeoted by BA only in older leaves. BA acts sequentially on the activities of photosynthetic and photorespiratory enzymes.     

Light-enhanced dark respiration in leaves and mesophyll protoplasts of pea in relation to photorespiration,respiration and some metabolites content

The respiration rate of leaves and mesophyll protoplasts of pea (Pisum sativum L.), from plants which were previously kept in darkness for 24 h was doubled following a period of photosynthesis at ambient level of O₂ (21 %), whereas the low level of O₂ (1 % and 4 % for leaves and protoplasts, respectively) reduced this light-enhanced dark respiration (LEDR) to the rate as noted before the illumination. Similarly to respiration rate, the oxygen at used concentrations had no effect on the ATP/ADP ratio in the dark-treated leaves. However, the ATP/ADP ratio in leaves photosynthesizing at 21 % O₂ was higher (up to 40 %, dependence on CO₂ concentration in the range 40–1600 1 dm⁻³) than in those photosynthesizing at 1 % O₂ or darkened at air (21 % O₂). Also, at 1 % O₂ the accumulation of malate was suppressed (by about 40 %), to a value noted for leaves darkened at 21 % O₂. The dark-treatment of leaves reduced the ability of isolated mitochondria to oxidize glycine (by about twofold) and succinate, but not malate. Mitochondria from both the light- and dark-treated leaves did not differ in qualitative composition of free amino acids, however, there were significant quantitative differences especially with respect to aspartate, alanine, glutamate and major intermediates of the photorespiratory pathway (glycine, serine). Our results suggest that accumulation of photorespiratory and respiratory metabolites in pea leaves during photosynthesis at 1 % O₂ is reduced, hence the suppression of postillumination respiration rate.