The occurrence of microbodies and peroxisomal enzymes in achlorophyllous leaves

Summary Several types of leaves of leaf parts lacking chlorophyll were fixed and embedded according to conventional procedures and examined electron-microscopically for microbodies. Comparisons of relative abundance of microbodies, plastids and mitochondria were made by computing the average numbers of organelle profiles per cell section. Similar leaves were homogenized and assayed for three enzymes characteristic of leaf peroxisomes. The localization of these enzymes in microbodies was indicated for the achlorophyllous tissues by the positive result obtained when 3,3-diaminobenzidine was used as an electron cytochemical stain for catalase activity.Microbodies were present in all non-photosynthetic leaves or leaf parts examined, including yellowish-white segments of variegated leaves, albino leaves, and etiolated leaves of two species. In several cases, the numbers of microbody profiles per cell section were as great in the achlorophyllous leaves as in the chlorophyllous. The levels of peroxisomal enzyme activity in the yellowish-white leaves were substantial, although often not as high as in the green leaves. It was concluded that enzymatically these microbodies are probably similar to the peroxisomes characterized from chlorophyllous leaves. In the absence of the photosynthetic product, glycolate, however, it seems unlikely that the organelle is performing the same functions as in green leaves. It is also apparent that the initial formation of peroxisomes in leaves can occur when neither light nor a photosynthate such as glycolate is present as an inducer.     

CO2 fixation in leaves of a CAM plant without lower epidermis and the effect of CO2 on their deacidification

The effects of peeling the epidermis off Bryophyllum daigremontianumleaves on CO₂ uptake in light and darkness were investigated.Light-induced CO₂ uptake in the daytime was markedly enhancedin the peeled leaves, but dark fixation of CO₂ carried out atmidnight was not. The difference in promotion of CO₂ uptakein light and darkness was due to stomatal closing in the dayand opening at night. Also, deacidification was strikingly inhibitedby CO₂ in peeled leaves. (Received February 3, 1977; )     

Ultrastructure of the vegetative cell ofBrassica napus pollen with particular reference to microbodies

Summary The ultrastructure of the vegetative cell ofBrassica napus tricellular pollen grains, just before anthesis with standard chemical fixation, is reported. The vegetative cell may be regarded as a highly differentiated and metabolically active fat-storage cell. It contains many mitochondria with a well developed internal membrane system, starchless plastids, microbodies, lipid bodies, dictyosomes and numerous vesicles thought to originate from the dictysomes. Rough endoplasmic reticulum organized in stacks of cisternae is also spatially associated with certain organelles, mainly lipid bodies, microbodies and plastids. There are also randomly distributed polyribosome areas. The microbodies are mainly polymorphic in shape and are often observed in contact with lipid bodies. The above spatial relationship implies that the microbodies may have a glyoxysomal function. In the late period of vegetative cell maturation, the microbodies are probably involved in the process of glyconeogenesis in which the conversion of lipid reserves to sugar takes place.Abbreviations VC vegetative cell - VN vegetative nucleus - SC sperm cell - M mitochondria - MB microbodies - L lipid body - P plastid - D dictyosomes     

Oligocene CO2 decline promoted C4 photosynthesis in grasses

C4 photosynthesis is an adaptation derived from the more common C3 photosynthetic pathway that confers a higher productivity under warm temperature and low atmospheric CO2 concentration [1, 2]. C4 evolution has been seen as a consequence of past atmospheric CO2 decline, such as the abrupt CO2 fall 32-25 million years ago (Mya) [3-6]. This relationship has never been tested rigorously, mainly because of a lack of accurate estimates of divergence times for the different C4 lineages [3]. In this study, we inferred a large phylogenetic tree for the grass family and estimated, through Bayesian molecular dating, the ages of the 17 to 18 independent grass C4 lineages. The first transition from C3 to C4 photosynthesis occurred in the Chloridoideae subfamily, 32.0-25.0 Mya. The link between CO2 decrease and transition to C4 photosynthesis was tested by a novel maximum likelihood approach. We showed that the model incorporating the atmospheric CO2 levels was significantly better than the null model, supporting the importance of CO2 decline on C4 photosynthesis evolvability. This finding is relevant for understanding the origin of C4 photosynthesis in grasses, which is one of the most successful ecological and evolutionary innovations in plant history.     

Mechanisms underlying CO2 diffusion in leaves

Plants provide an excellent system to study CO(2) diffusion because, under light saturated conditions, photosynthesis is limited by CO(2) availability. Recent findings indicate that CO(2) diffusion in leaves can be variable in a short time range. Mesophyll CO(2) conductance could change independently from stomata movement or CO(2) fixing reactions and it was suggested that, beside others, the membranes are mesophyll CO(2) conductance limiting components. Specific aquaporins as membrane intrinsic pore proteins are considered to have a function in the modification of membrane CO(2) conductivity. Because of conflicting data, the mechanism of membrane CO(2) diffusion in plants and animals is a matter of a controversy vivid debate in the scientific community. On one hand, data from biophysics are in favor of CO(2) diffusion limiting mechanisms completely independent from membrane structure and membrane components. On the other, there is increasing evidence from physiology that a change in membrane composition has an effect on CO(2) diffusion.     

Responses of C4 grasses to atmospheric CO2 enrichment

Summary The growth and photosynethetic responses to atmospheric CO₂ enrichment of 4 species of C₄ grasses grown at two levels of irradiance were studied. We sought to determine whether CO₂ enrichment would yield proportionally greater growth enhancement in the C₄ grasses when they were grown at low irradiance than when grown at high irradiance. The species studied were Echinochloa crusgalli, Digitaria sanguinalis, Eleusine indica, and Setaria faberi. Plants were grown in controlled environment chambers at 350, 675 and 1,000 l 1^-1 CO₂ and 1,000 or 150 mol m^-2 s^-1 photosynthetic photon flux density (PPFD). An increase in CO₂ concentration and PPFD significantly affected net photosynthesis and total biomass production of all plants. Plants grown at low PPFD had significantly lower rates of photosynthesis, produced less biomass, and had reduced responses to increases in CO₂. Plants grown in CO₂-enriched atmosphere had lower photosynthetic capacity relative to the low CO₂ grown plants when exposed to lower CO₂ concentration at the time of measurement, but had greater rate of photosynthesis when exposed to increasing PPFD. The light level under which the plants were growing did not influence the CO₂ compensation point for photosynthesis.     

Effects of elevated CO2 on competition between native and invasive grasses

Oecologia - Elevated atmospheric CO2 concentration increases the performance of invasive plants relative to natives when grown in monoculture, but it is unclear how that will affect the relative...     

Ultrastructure and isolation of glyoxysomes (microbodies) in zoospores of the fungusentophlyctis sp.

Summary A correlative approach, involving light and electron microscopic, cytochemical, and biochemical techniques, was used to study the structure and function of microbodies in zoospores ofEntophlyctis sp. The same population of microbodies already existing in the zoosporangium appeared to be segregated into zoospore initials during cytoplasmic cleavage. Microbodies laid at the anterior end of zoospores and were part of an organized assemblage of organelles, the microbody-lipid globule complex. In the microbody-lipid globule complex, endoplasmic reticulum occurred on the surface of the lipid globules toward the zoospore's exterior, and the microbody, subtended by mitochondria, was appressed to the opposite surface of the lipid globule. The organization of the microbody-lipid globule complex changed as the zoospore swam and encysted. As lipid globules coalesced, the microbody-lipid globule complex became disorganized. After lipid globule coalescence was completed, the microbody-lipid globule complex regained its order, and several microbodies were clustered adjacent to a single lipid globule. The microbodies persisted even in the encysted zoospore, but they were found on all sides of the lipid globule.Microbodies isolated from zoospores contained catalase as well as malate synthase and isocitrate lyase, two enzymes of the glyoxylate cycle. When zoospores encysted greater activities of these glyoxylate cycle enzymes could be detected. The presence of glyoxylate cycle enzymes and the close association between the microbody and lipid globule suggest that microbodies function as glyoxysomes in zoospores and encysted zoospores. The functional significance of the morphological organization of the microbody-lipid complex is discussed in terms of energy production and the conversion of storage lipid into structural components of the cell.     

A eukaryote without catalase-containing microbodies: Neurospora crassa exhibits a unique cellular distribution of its four catalases

Microbodies usually house catalase to decompose hydrogen peroxide generated within the organelle by the action of various oxidases. Here we have analyzed whether peroxisomes (i.e., catalase-containing microbodies) exist in Neurospora crassa. Three distinct catalase isoforms were identified by native catalase activity gels under various peroxisome-inducing conditions. Subcellular fractionation by density gradient centrifugation revealed that most of the spectrophotometrically measured activity was present in the light upper fractions, with an additional small peak coinciding with the peak fractions of HEX-1, the marker protein for Woronin bodies, a compartment related to the microbody family. However, neither in-gel assays nor monospecific antibodies generated against the three purified catalases detected the enzymes in any dense organellar fraction. Furthermore, staining of an N. crassa wild-type strain with 3,3'-diaminobenzidine and H(2)O(2) did not lead to catalase-dependent reaction products within microbodies. Nonetheless, N. crassa does possess a gene (cat-4) whose product is most similar to the peroxisomal type of monofunctional catalases. This novel protein indeed exhibited catalase activity, but was not localized to microbodies either. We conclude that N. crassa lacks catalase-containing peroxisomes, a characteristic that is probably restricted to a few filamentous fungi that produce little hydrogen peroxide within microbodies.     

Postillumination CO2 fixation by wheat leaves

Postillumination CO₂ fixation by wheat leaves was studied following light-limited photosynthetic conditions. Dark CO₂ fixation showed two phases differing by their rates of CO₂ uptake and carbon metabolism. These two phases are related to preillumination light flux density. During the first 30s of darkness, assimilated CO₂ was found in PGA, alanine, malate and aspartate. After 5 min of darkness, it was additionally found in phosphorylated sugars.The lack of labelling of glycolate pathway intermediates shows that the Calvin cycle cannot run in the dark.The synthesized compounds indicate that reducing power but not ATP is available after turning the light off. This observation suggests that during pre-illumination, when light strictly limits photosynthesis, ATP supply would be the first limiting factor.

---

Résumé La fixation post-illuminatoire de CO₂ par des feuilles de blé est étudiée, après une période de photosynthèse en lumière limitante.Les cinétiques de la vitesse de fixation du CO₂ après extinction présentent deux phases, se différenciant par la vitesse de fixation du CO₂ et par les voies métaboliques suivies par le carbone.Pendant les premières 30s d'obscurité, le CO₂ fixé est retrouvé principalement dans le PGA, l'alanine, le malate et l'aspartate. Après 5 min d'obscurité le carbone est retrouvé également dans les esters phosphorylés des oses.L'absence de radioactivité dans les intermédiaires de la voie du glycolate indique que le cycle de Calvin ne peut pas fonctionner á l'obscurité.Les composés synthétisés à l'obscurité suggèrent que du pouvoir réducteur est disponible. Par contre l'ATP ne l'est pas. Ainsi, durant la période de pré-illumination oú la lumière était strictement limitante la disponibilité en ATP serait le premier facteur limitant l'assimilation du CO₂.

     

The quantum yield for CO2 uptake in C3 and C4 grasses

The quantum yield for CO₂ uptake was measured in C₃ and C₄ monocot species from several different grassland habitats. When the quantum yield was measured in the presence of 21% O₂ and 340 cm³ m^-3 CO₂, values were very similar in C₃ monocots, C₃ dicots, and C₄ monocots (0.045–0.056 mole CO₂ · mole^-1 quanta absorbed). In the presence of 2% O₂ and 800 cm³ m^-3 CO₂, enhancements of the quantum yield values occurred for the C₃ plants (both monocots and dicots), but not for C₄ monocots. A dependence of the quantum yield on leaf temperature was observed in the C₃ grass, Agropyron smithii, but not in the C₄ grass, Bouteloua gracilis, in 21% O₂ and 340 cm³ m^-3 CO₂. At leaf temperatures between 22–25°C the quantum yield values were approximately equal in the two species.     

Ultrastructure of methanol-utilizing yeast cells: appearance of microbodies in relation to high catalase activity.

Nine strains of methanol-utilizing yeasts belonging to the genera Candida, Hansenula, Kloeckera, Pichia, and Torulopsis were examined with respect to the interrelationship between their catalase content and ultrastructure. Methanol-grown cells of all the yeasts tested showed higher catalase activities than the respective ethanol- and glucose-grown cells. In connection with this, occurrence of a specific organelle surrounded by a single-unit membrane ("microbodies") was observed only in the methanol-grown cells. Several morphological differences were observed between the microbodies of methanol-utilizing yeasts and those of hydrocarbon-utilizing yeasts such as Candida tropicalis. That is, microbodies of methanol utilizers were large in size, existed in closely associated forms, and had crystalloid structures. Localization of catalase activity in these microbodies was demonstrated cytochemically by use of 3,3'-diaminobenzidene. Especially, 3,3'-diaminobenzidine reaction product accumulated heavily in crystalloids of yeast microbodies.     

Effect of carbon dioxide and temperature on photosynthetic CO2 uptake and photorespiratory CO2 evolution in sunflower leaves

Using an open gas-exchange system, apparent photosynthesis, true photosynthesis (TPS), photorespiration (PR) and dark respiration of sunflower (Helianthus annuus L.) leaves were determined at three temperatures and between 50 and 400 l/l external CO₂. The ratio of PR/TPS and the solubility ratio of O₂/CO₂ in the intercellular spaces both decreased with increasing CO₂. The rate of PR was not affected by the CO₂ concentration in the leaves and was independent of the solubility ratio of oxygen and CO₂ in the leaf cell. At photosynthesis-limiting concentrations of CO₂, the ratio of PR/TPS significantly increased from 18 to 30°C and the rate of PR increased from 4.3 mg CO₂ dm^-2 h^-1 at 18°C to 8.6 mg CO₂ dm^-2 h^-1 at 30°C. The specific activity of photorespired CO₂ was CO₂-dependent but temperature-independent, and the carbon traversing the glycolate pathway appeared to be derived both from recently fixed assimilate and from older reserve materials. It is concluded that PR as a percentage of TPS is affected by the concentrations of O₂ and CO₂ around the photosynthesizing cells, but the rate of PR may also be controlled by other factors.Abbreviations APS apparent photosynthesis (net CO₂ uptake) - PR photorespiration (CO₂ evolution in light) - RuBP ribulose-1,5-bisphosphate - TPS true photosynthesis (true CO₂ uptake)     

The relationship between CO2 assimilation and electron transport in leaves

The inter-relationships between the quantum efficiencies of photosystems I (_I) and II (_II) and the quantum yield of CO₂ fixation % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabaGaciaacaqabeaadaqaaqaaaOqaaiabeA8aMnaaBa% aaleaacaWGdbGaam4tamaaBaaameaacaaIYaaaleqaaaqabaaaaa!3BD3!\[\phi _{CO_2 } \] were investigated in pea (Pisum sativum (L)) leaves with differing rates of photosynthesis using both photorespiratory and non-photorespiratory conditions, and in a leaf of Hedera helix (L) under photorespiratory conditions. The results indicate that under photorespiratory conditions the relationship between % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabaGaciaacaqabeaadaqaaqaaaOqaaiabeA8aMnaaBa% aaleaacaWGdbGaam4tamaaBaaameaacaaIYaaaleqaaaqabaaaaa!3BD3!\[\phi _{CO_2 } \] and both _I and _II is non-linear and variable. The relationship between _I and _II under these circumstances remains predominantly linear. Under non-photorespiratory conditions, leaves with a low rate of photosynthesis due to sink limitation exhibit a non-linear relationship between _I and _II, though the relationship between _I and _II remains linear suggesting a close relationship between linear electron flow and CO₂ fixation. Leaves irradiated at the CO₂ compensation point also exhibit a non-linear relationship between _I and _II. These results suggest that for leaves in air linear electron flow is the predominant source of energy for metabolism. The role of cyclic electron transport is considered when the requirement for the products of linear electron transport is depressed.Abbreviations q_p the coefficient for photochemical quenching of chlorophyll fluorescence - _exe the quantum efficiency of excitation energy capture by open PS II traps - _II the quantum efficiency for electron transport by PS II - _I the quantum efficiency (for electron transport) by PS I - % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabaGaciaacaqabeaadaqaaqaaaOqaaiabeA8aMnaaBa% aaleaacaWGdbGaam4tamaaBaaameaacaaIYaaaleqaaaqabaaaaa!3BD3!\[\phi _{CO_2 } \] the quantum yield for CO₂ fixation (obtained as the gross rate of CO₂ fixation divided by the irradiance) - % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabaGaciaacaqabeaadaqaaqaaaOqaaiabgs5aenaaBa% aaleaacqaH8oqBdaWgaaadbaGaamisamaaCaaabeqaaiabgUcaRaaa% aeqaaaWcbeaaaaa!3CB0!\[\Delta _{\mu _{H^ + } } \] trans-thylakoid proton potential difference - PAQF photosynthetically active quantum flux     

Emissions of nitrous oxide from the leaves of grasses

Aims

Nitrous oxide (N₂O) emissions from pastoral agriculture are considered to originate from the soil as a consequence of microbial activity during soil nitrification and denitrification. However, recent studies have identified the plant canopy as a potentially significant source of N₂O emissions to the atmosphere. Understanding the extent and mechanisms of plant emissions may provide new mitigation opportunities as current options only target soil microbial processes.

Methods

We developed an experimental apparatus and protocol to partition N₂O emissions between the leaves of grasses and the soil and measured emissions from ten common grass species found in New Zealand pastures.

Results

The chamber design enabled us to identify measurable changes in N₂O concentration over a period of 1 h and to distinguish a range of emissions from 0.001 to 0.25 mg N₂O-N/m² leaf area/h. There was a 10-fold variation among species; Holcus lanataus, Lolium perenne and Paspalum dilatatum had the highest leaf N₂O emissions and Poa annua the lowest.

Conclusions

Grasses do emit N₂O from their leaves and the rate that this occurs varies among grass species. The emission does not appear to arise from formation of N₂O in plant leaves but more likely reflects transport of N₂O from the soil. Differences in emission rates appear to arise from a plant influence on the rate of formation of N₂O in the soil rather than the rate of transportation through the plant.