Dissimilar phloem loading in leaves with symplasmic or apoplasmic minor-vein configurations

Plant species were selected on the basis of abundant or no symplasmic continuity between sieveelement-companion-cell (SE-CC) complexes and adjacent cells in the minor veins. Symplasmic continuity and discontinuity are denoted, respectively, as symplasmic and apoplasmic minor-vein configurations. Discs of predarkened leaves from which the lower epidermis had been removed, were exposed to ¹⁴CO₂. After 2 h of subsequent incubation, phloem loading in control discs and discs treated with p-chloromercuribenzenesulfonic acid (PCMBS) was recorded by autoradiography. Phloem loading was strongly suppressed by PCMBS in minor veins with symplasmically isolated SE-CC complexes (Centaurea, Impatiens, Ligularia, Pelargonium, Pisum, Symphytum). No significant inhibition of phloem loading by PCMBS was observed in minor veins containing sieve elements with abundant symplasmic connections (Epilobium, Fuchsia, Hydrangea, Oenothera, Origanum, Stachys). Phloem loading in minor veins with both types of SE-CC complex (Acanthus) had apoplasmic features. The results provide strong evidence for coincidence between the mode of phloem loading and the minor-vein configuration. The widespread occurrence of a symplasmic mode of phloem loading is postulated.Abbreviations PCMBS p-chloromercuribenzenesulfonic acid - SE-CC complex sieve-element-companion-cell complex     

Phloem loading and its development related to plant evolution from trees to herbs

Summary Minor vein structure in various taxonomic groups was described in a previous paper (Gamalei 1989). Here, these results are used to correlate minor vein structure with plant evolutionary, ecological and growth form schemes. The following pattern emerges: reductive evolution from evergreen trees to annual herbs is accompanied by gradually increasing symplastic isolation of the mesophyll and the phloem. This evolutionary tendency is confirmed by the ecological spreading and life-form distribution of modern plants with different types of minor vein structure. The meaning of this phenomenon is discussed. Chilling sensitivity of plasmodesmal translocation is considered to be the main reason. It is suggested that phloem loading for assimilate transport is double-routed. The symplastic route is more ancient and more economical for loading. The apoplastic pathway becomes the main or the only route under unfavorable conditions. The existence of a symplast/apoplast regulatory loading mechanism is suggested. The two loading routes differ in their selectivity for products of photosynthesis which changes their symplast/apoplast ratio which, in turn, determines the composition of the sieve tube exudate. The latter will influence growth and morphogenesis. Correlated changes of structure and function related to photosynthesis, loading, translocation and growth, are analysed with respect to life-form evolution. The influence of the pathway of loading on other processes is discussed.     

The Role of Plastids and Assimilate Transport System in the Control of Plant Development

Phylogenetic and ontogenetic relationships between the plastids, cell endoplasmic reticulum, and plant transport communication have been reviewed. The initiating role of plastids (endosymbionts) in the origin of endoplasmic reticulum (buffer zone of endosymbiogenesis) has been shown, as well as a similar role of endoplasmic reticulum in the development of transport communication of xylem and phloem. Plastids, sugars and transport system for their distribution can be interpreted as leading sections in the mechanism of developmental control: gene expression of nuclear genome and genome of organelles, cell and tissue differentiation, and plant morphogenesis. The conflict between the bulk of plant genome and low percentage of its realization is explained as a result of limitation of the nuclear genome realization by plastid genome. The concept of development as applied to plant ontogenesis has been critically analyzed. The possibilities of the concept correction by with the help of symbiogenetic hypothesis are discussed.__________Translated from Ontogenez, Vol. 36, No. 3, 2005, pp. 165–181.Original Russian Text Copyright © 2005 by Gamalei.     

Energized Uptake of Sugars from the Apoplast of Leaves: A Study of Some Plants Possessing Different Minor Vein Anatomy

Solutions of sucrose, glucose, raffinose, and stachyose were fed via the petiole to detached leaves of plant species known to transfer sugars during photosynthesis into the phloem using either the apoplastic or the symplastic pathway of phloem loading. Symplastic phloem loaders, which translocate raffinose-type oligosaccharides and sucrose in the phloem, and apoplastic plants, translocating exclusively sucrose, were selected for this study. As the sugars arrived with the transpiration stream in the leaf blade within little more than a minute, dark respiration increased. Almost simultaneously, fluorescence of a potential-indicating dye, which had been infiltrated into the leaves, indicated membrane depolarization. Another fluorescent dye used to record the apoplastic pH revealed apoplastic alkalinization that occurred with a slight lag phase after respiration and membrane depolarization responses. Occasionally, alkalinization was preceded by transient apoplastic acidification. Whereas membrane depolarization and apoplastic acidification are interpreted as initial responses of the proton motive force across the plasma membrane to the advent of sugars in the leaf apoplast, the following apoplastic alkalinization showed that sugars were taken up from the apoplast into the symplast in cotransport with protons. This was true not only for glucose and sucrose, but also for raffinose and stachyose. Similar observations were made for sugar uptake not only in leaves of plants known to export sugars by symplastic phloem loading but also of plants using the apoplastic pathway. Increased respiration during sugar uptake revealed tight coupling between respiratory ATP production and ATP consumption by proton-translocating ATPase of the plasma membrane, which exports protons into the apoplast, thereby compensating for the proton loss in the apoplast when protons are transported together with sugars into the symplast. The extent of stimulation of respiration by sugars indicated that sugar uptake was not limited to phloem tissue. Ratios of the extra CO₂ released during sugar uptake to the amounts of sugars taken up were variable, but lowest values were lower than 0.2. When a ratio of 0.2 is taken as a basis to calculate rates of sugar uptake from observed maxima of sugar-dependent increases in respiration, rates of sugar uptake approached 350 nmol/(m² leaf surface s). Sugar uptake rates were half-saturated at sugar concentrations in the feeding solutions of about 10–25 mM indicating a low in vivo affinity of sugar uptake systems for sugars.     

Photooxidative Damage in Young Leaves of Declining Grapevine: Does It Result from a New and Possibly Viroid-Related Disease?

Leaves of chlorotic plants of Vitis vinifera were investigatedfor physiological and ultrastructural disorders by comparingthem with leaves of apparently healthy plants from a vineyard,where infection with Hop Stunt Viroid F (HSVdg) and GrapevineYellow Speckle Viroid I (GYSVdl) was widespread. In affectedplants, chlorosis was much stronger in young and developingthan in old and fully expanded leaves. Chemical analyses failedto reveal mineral deficiencies. The quantum yield of photosynthesiswas decreased in chlorotic leaves. A decrease in the numberof PSI reaction centers was also observed. Persisting photoinhibitionoccurred only in leaves of affected plants. The redox stateof cellular and extracellular ascorbate and increased levelsof glutathione indicated oxidative stress in affected plants.Ultrastructural analysis revealed both swelling and loss ofthylakoids even in young chlorotic leaves and other pathologicalchanges. Symptoms were similar to those normally observed onlyin old senescing leaves. However, chlorotic leaves showed unexpectedlyhigh protein levels, though aging is known to lead to proteindegradation. (Received April 30, 1996; Accepted October 14, 1996)     

Plant cell systems

Our present view of the structure of higher plant cell systems arising on the basis of electron, fluorescent, and confocal microscopy and also video movies of living cells under confocal microscope is presented. The role of the partition (conducting) system in the arrangement of the whole plant cell systems, their development, breakdown, and renewal is demonstrated. The phylogenetic origination of the partition net and its primary role in the ontogenetic development of the cell systems are considered in the context of the theory of endosymbiogenesis. The data concerning growth limits and a diversity of the higher plant cell system organization are compared. The specific features of the cell systems of prokaryotes, autotrophic and heterotrophic eukaryotes, and also of their evolution are discussed.     

Effects of temperature on the conformation of the endoplasmic reticulum and on starch accumulation in leaves with the symplasmic minor-vein configuration

The phloem-loading-related effects of temperature on leaf ultrastructure were studied in seven species having numerous plasmodesmatal connections between the mesophyll and phloem (symplasmic minor-vein configuration). The response to temperature (between 5 and 30 °C) was characterized by drastic changes in the endoplasmic-reticulum labyrinth (ER labyrinth) of intermediary cells, in the position of the vacuole in bundle-sheath cells, and in the starch content in the chloroplasts of bundle-sheath cells and mesophyll cells. At temperatures above 20 °C, the ER system in the intermediary cells reached its maximal volume, while the vacuole in bundlesheath cells was positioned centripetally (proximal to the intermediary cell). With decreasing temperature, the ER labyrinth in intermediary cells gradually contracted till the ER was fully collapsed at 10 °C and the vacuole in bundle-sheath cells moved to a more centrifugal position. The apparent elimination of photosynthate transport via the ER and plasmodesmata at temperatures lower than 10 °C led to starch accumulation in the chloroplasts of bundle-sheath cells and mesophyll cells. All of these changes were fully temperature-reversible and probably reflect changes in the balance between photosynthate transport and storage. The ultrastructural shifts appear to be correlated with the passage of photosynthate through the intermediary cells and, as a consequence, with the rate of phloem loading at various temperatures. A contraction of the ER/plasmodesmata system imposed by cytoskeletal reorganisation is discussed as the reason for the blockage of phloem loading at low temperatures in association with the general chilling sensitivity of these species.Abbreviations BSC bundle-sheath cell - IC intermediary cell - MC mesophyll cell - PD plasmodesmata - PFD photon flux density - SE/CC-complex sieve element/companion cell complex The authors gratefully acknowledge the financial support by NWO (Dutch Organization for Scientific Research).     

Phloem loading in two Scrophulariaceae species. What can drive symplastic flow via plasmodesmata?

To determine the driving forces for symplastic sugar flux between mesophyll and phloem, gradients of sugar concentrations and osmotic pressure were studied in leaf tissues of two Scrophulariaceae species, Alonsoa meridionalis and Asarina barclaiana. A. meridionalis has a typical symplastic configuration of minor-vein phloem, i.e. intermediary companion cells with highly developed plasmodesmal connections to bundle-sheath cells. In A. barclaiana, two types of companion cells, modified intermediary cells and transfer cells, were found in minor-vein phloem, giving this species the potential to have a complex phloem-loading mode. We identified all phloem-transported carbohydrates in both species and analyzed the levels of carbohydrates in chloroplasts, vacuoles, and cytoplasm of mesophyll cells by nonaqueous fractionation. Osmotic pressure was measured in single epidermal and mesophyll cells and in whole leaves and compared with calculated values for phloem sap. In A. meridionalis, a 2-fold concentration gradient for sucrose between mesophyll and phloem was found. In A. barclaiana, the major transported carbohydrates, sucrose and antirrhinoside, were present in the phloem in 22- and 6-fold higher concentrations, respectively, than in the cytoplasm of mesophyll cells. The data show that diffusion of sugars along their concentration gradients is unlikely to be the major mechanism for symplastic phloem loading if this were to occur in these species. We conclude that in both A. meridionalis and A. barclaiana, apoplastic phloem loading is an indispensable mechanism and that symplastic entrance of solutes into the phloem may occur by mass flow. The conditions favoring symplastic mass flow into the phloem are discussed.