Phloem water relations and translocation

Satisfactory measurements of phloem water potential of trees can be obtained with the Richards and Ogata psychrometer and the vapor equilibration techniques, although corrections for loss of dry weight and for heating by respiration are required for the vapor equilibrium values. The psychrometer technique is the more satisfactory of the 2 because it requires less time for equilibration, less tissue, and less handling of tissue. Phloem water potential of a yellow-poplar tree followed a diurnal pattern quite similar to that of leaves, except that the values were higher (less negative) and changed less than in the leaves.

The psychrometer technique permits a different approach to the study of translocation in trees. Measurements of water potential of phloem discs followed by freezing of samples and determination of osmotic potential allows estimation of turgor pressure in various parts of trees as the difference between osmotic potential and total water potential. This technique was used in evaluating gradients in water potential, osmotic potential, and turgor pressure in red maple trees. The expected gradients in osmotic potential were observed in the phloem, osmotic potential of the cell sap increasing (sap becoming more dilute) down the trunk. However, values of water potential were such that a gradient in turgor pressure apparently did not exist at a time when rate of translocation was expected to be high. These results do not support the mass flow theory of translocation favored by many workers.

     

Mechanism of cyanide inhibition of Phloem translocation

Petiolar application of potassium cyanide inhibited ¹⁴C-assimilate translocation without affecting source leaf photosynthesis or phloem loading of sucrose in Phaseolus vulgaris. The inhibition of transport was correlated with disruption of the structural integrity of the sieve tubes (sieve pore blockage) rather than impairment of a metabolic process in the translocation path driving translocation.     

Phloem translocation of gibberellins in three species of higher plants

Phloem sap was collected from white lupin (Lupinus albus L.), cowpea (Vigna unguiculata L.) and castor bean (Ricinus communis L.) and analysed for gibberellins (GAs) using gas chromatography-mass spectrometry (GC-MS). A large number of GAs were found in the phloem exudate of all three species, particularly where the sap was collected from pods (white lupin and cowpea) and in these legumes GAs representing both the early C-13-hydroxylation and non-hydroxylation pathways of biosynthesis were identified. In the sap collected from the vegetative tissues of castor bean the number of GAs identified was fewer than that in the other species, representing mainly the non-hydroxylation pathway. Data from sap collected from the pedicel and stylar ends of pods and by making feeds of radiolabelled GAs to seeds in situ in white lupin indicate that the GAs present in the phloem are derived mainly from the vegetative tissues of the plant. No evidence for metabolism of GAs in the phloem could be found.     

Phloem translocation in regenerating in vitro- heterografts of different compatibility

Phloem translocation of [¹⁴C]-sucrose and 5/6-carboxyfluorescein(CF) from scion into the stock was studied in in vitro-heterograftsof Lycopersicon on Solanum (L/S) and Vicia on Helianthus (V/H)at various stages of regeneration. Autografts of all partnersserved as controls. Corresponding with the translocation experimentsnewly formed sieve-tube connections between the graft partnerswere counted. ¹⁴C-translocation experiments with [¹⁴C]-sucrose revealed anage-dependent increase of radioactivity in the stock of allcombinations. In L/S and all autografts the major increase of¹⁴C-label in the stock occurred 5–10 d after grafting.In V/H, however, import of label into the stock remained lowthroughout the regeneration period. In L/S grafts, increasesin the numbers of sieve-tube connections parallel the increasingrate of ¹⁴C-transport, indicating functioning sieve-tube connectionsin the graft union. In contrast, V/H grafts did not show thisstrong correlation between structure and function of wound repairphloem. This suggested the existence of non-transporting sieve-tubesbetween the graft partners. Similar results were obtained withCF-transport, showing that effective phloem translocation acrossthe graft interface occurred in L/S, but not in V/H grafts.The observed differences in phloem translocation are discussedwith regard to compatibility/incompatibility phenomena in heterografts. Key words: Compatibility/incompatibility, in-vitro-heterografts, phloem transport ([¹⁴C]-sucrose, carboxyfluorescein), wound phloem     

Phloem translocation of a reduced oligogalacturonide inRicinus communis L

We have re-examined the evidence against the phloem mobility of oligogalacturonide elicitors using a reduced oligogalacturonide in the phloem translocation system ofRicinus communis var. Gibsonii. A tritium-labelled end-reduced oligogalacturonide of degree of polymerisation 6 was injected into the hollow centre of the petiole of four- to five-week-old plants. Two experimental procedures were followed. In the first, the whole plant was harvested and dissected after 5 h incubation. In the second, phloem sap was collected from an incision in the main stem below the injected petiole; collection started 2 h after incubation and continued for a further 3 h. Determination of the total radiolabel present in the dissected plant showed that at least 8% of the applied activity was exported from the injected leaf, most of this being recovered from the main stem below the injected petiole and the roots. The activity in the phloem exudate showed that the rate of export of radiolabel was already at its maximum by the end of the 2-h incubation period. Radiolabelled material recovered from the main stem was found to be highly comparable to starting material when subjected to thin-layer chromatography. These results demonstrate the phloem mobility of reduced oligogalacturonides of low degree of polymerisation and therefore re-establish the potential for oligogalacturonides to act as systemic signals.     

An investigation of the contractile protein hypothesis of phloem translocation

Summary Experiments are reported which were designed to test the hypothesis that the movement of the translocation stream is driven by the contractile activity of P-protein filaments. The different types of filament found after negative staining of phloem exudates from Ricinus communis and Cucurbita pepo are described. An approximate model is proposed for the quaternary structure of a 20 nm component in the R. communis exudate. None of the filaments showed any ability to bind heavy meromyosin subfragment one. In experiments with cytochalasin B, no evidence of effects on the movement of ¹⁴C-assimilates or on the ultrastructure of the sieve elements of Lepidium sativum was found. It is concluded that the available evidence is unfavourable to the view that P-protein resembles known contractile proteins elsewhere.     

Microautoradiographic investigations on bidirectional translocation in the phloem of Vicia faba

Summary Using radioactive phenylalanine as a tracer, bidirectional translocation in the phloem of a single stem bundle of Vicia faba L. was detected by analyses of microautoradiographs. Bidirectional translocation occurred in lateral leaf traces subtending a leaf of a certain developmental stage. Before and after this stage, translocation was unidirectional, either acropetal or basipetal, respectively.     

Solute distribution in sugar beet leaves in relation to Phloem loading and translocation

The distribution of solutes in the various cells of sugar beet (Beta vulgaris L.) source leaves, petioles, and sink leaves was studied in tissue prepared by freeze-substitution. The differences in degree of cryoprotection indicated that sieve elements and companion cells of the source leaf, petiole, and sink leaf contain a high concentration of solute. The osmotic pressure of various types of cells was measured by observing incipient plasmolysis in freeze-substituted tissues equilibrated with a series of mannitol solutions prior to rapid freezing. Analysis of source leaf tissue revealed osmotic pressure values of 13 bars for the mesophyll and 30 bars for the sieve elements and companion cells. The osmotic pressure of the mesophyll of sink leaves was somewhat higher.     

Phloem ultrastructure and pressure flow: Sieve-Element-Occlusion-Related agglomerations do not affect translocation

Since the first ultrastructural investigations of sieve tubes in the early 1960s, their structure has been a matter of debate. Because sieve tube structure defines frictional interactions in the tube system, the presence of P protein obstructions shown in many transmission electron micrographs led to a discussion about the mode of phloem transport. At present, it is generally agreed that P protein agglomerations are preparation artifacts due to injury, the lumen of sieve tubes is free of obstructions, and phloem flow is driven by an osmotically generated pressure differential according to Münch's classical hypothesis. Here, we show that the phloem contains a distinctive network of protein filaments. Stable transgenic lines expressing Arabidopsis thaliana Sieve-Element-Occlusion-Related1 (SEOR1)-yellow fluorescent protein fusions show that At SEOR1 meshworks at the margins and clots in the lumen are a general feature of living sieve tubes. Live imaging of phloem flow and flow velocity measurements in individual tubes indicate that At SEOR1 agglomerations do not markedly affect or alter flow. A transmission electron microscopy preparation protocol has been generated showing sieve tube ultrastructure of unprecedented quality. A reconstruction of sieve tube ultrastructure served as basis for tube resistance calculations. The impact of agglomerations on phloem flow is discussed.     

Source and sink leaf metabolism in relation to Phloem translocation: carbon partitioning and enzymology

The import-export transition in sugar beet leaves (Beta vulgaris) occurred at 40 to 50% leaf expansion and was characterized by loss in assimilate import and increase in photosynthesis. The metabolism and partitioning of assimilated and translocated C were determined during leaf development and related to the translocation status of the leaf. The import stage was characterized by C derived from either ¹⁴C-translocate or ¹⁴C-photosynthate being incorporated into protein and structural carbohydrates. Marked changes in the C partitioning were temporally correlated with the import-export conversion. Exporting leaves did not hydrolyze accumulated sucrose and the C derived from CO₂ fixation was preferentially incorporated into sucrose. Both source and sink leaves contained similar levels of acid invertase and sucrose synthetase activities (sucrose hydrolysis) while sucrose phosphate synthetase (sucrose synthesis) was detected only in exporting leaves. The results are discussed in terms of intracellular compartmentation of sucrose and sucrose-metabolizing enzymes in source and sink leaves.     

An Aphid-Secreted Salivary Protease Activates Plant Defense in Phloem

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Solution-Flow in the Phloem: II. Phloem Transport of THO in Beta vulgaris

Translocation profiles along the path were studied using a modified flap-feeding technique for the simultaneous application of THO and ¹⁴C-sucrose. A re-evaluation of a mathematical model for phloem transport with reversible lateral exchange of tracer along the path indicates that lower apparent velocities for THO as compared to labeled carbohydrate are primarily due to extensive lateral exchange of THO along the conduction path. Path-chilling experiments support the concept that THO and ¹⁴C-sucrose exhibit different lateral exchange characteristics. The data presented are consistent with a solutionflow mechanism.     

Tension in the Phloem?

The hydrostatic pressure in sieve tubes may probably be estimatedreasonably accurately from experimental measurements of theosmolarity of the sieve tube sap and of the tension in the xylem.The possibility of the existence of phloem tension is advanced.     

Anatomy of the Secondary Phloem and the Crystalliferous Phloem Fibers in the Stem of Torreya grandis

The structure of the secondary phloem and the development of the crystaleiferous phloem fibers in the stem of Torrey grandis were observed under the ligth microscope and SEM. The secondary phloem is composed of sieve cells, phloem parenchyma cells, crystalliferous phloem fibers and stone cells in the longitudinal system, and the uniserite homogeneous phloem rays consisting of parenchyma cells only in the radial system. In the cross section, there are 3–9 sieve cells in radial rows forming discontinuous tangential layers, the crystalliferous phloem fibers often in a single discontinuous tangential layer and the stone cells dispersed in rangential layer of phloem parenchyma. The developmental process of crystalliferous phloem fibers is as follows: initial cells appeared in the end of April and were well differentiated in the first week of May. Some crystals were deposited in the primary wall, while others were free in the cell. At the end of May, the secondary wall of most crysalliferous phloem fibers started to be thickened. With the thickening of the secondary wall, all the crystals were embedded in the wall from June to August From the end of September to the early days of October, the crystalliferous phloem fibers reached their full maturation. It is shown by microchemical identification and EDAX analysis that the crystals embedded in the wail of crystalliferous phloem fibers are calcium oxalate crystals.     

An Investigation of the Relay Hypothesis of Phloem Transport in Ricinus communis L.

To test for the existence of an apoplastic unloading/reloadingstep in phloem translocation, as envisaged in the relay hypothesisof phloem transport, isotopic trapping experiments were performedon Ricinus comunis L. var. Gibsonii. A CO₂ buffer system wasused to supply ¹⁴CO₂ at constant partial pressure and constantspecific activity to a photosynthesizing leaf. The subtendinginternode was perfused with solutions of sugars or of mannitoland transolation of ¹⁴C past the perfused zone was monitoredby the collection of phloem exudate. Trapping of activity inthe perfusate was enhanced by the presence of sugars as wouldbe expected with an unloading/reloading process. However, therewas no evidence that introduction of the unlabelled sugars tothe apoplast also reduced activity in the phloem exudate. Moreover,the rate of loss of activity to the perfusing solutions representedonly 1% of the rate of appearance in the exudation. It is suggestedthat the trapping results may reflect an unloading of tracerfrom the phloem associated with a subsequent reloading by adjacenttissues rather than by the sieve tubes. To investigate the length of sieve tube continuity in Ricinus,a horizontal incision was made to the bark and the rate of exudationof phloem sap was monitored. Successive circumferential cutswere made above the exuding incision and progressively closerto it. In general, a girdling incision produced a transientdecrease in the rate of change of exudation rate (i.e. the firstderivative became more negative/less positive). The magnitudeof this response rose with exudation rate and fell with thedistance at which a girdling cut was made. Fitting an appropriatemodel yielded an estimate for contributory length of 69 ±6 cm. This was comparable with the distance of the initial tangentialincision from the stem apex, suggesting a continuous sieve tubesystem in Ricinus. A similar investigation on the petiole yieldedan estimate of around 7.0 cm. This lower estimate for contributorylength is believed to reflect a rapid sealing process that limitsthe distance of propagation of turgor-release rather than alimited length of sieve tube continuity. The results of this investigation do not support the relay hypothesisof phloem transport. Rather they suggest a continuous sievetube system which has a distributed capacity to load and unloadsolutes, and which may exhibit a sealing response when injured. Key words: Ricinus communis L, phloem transport, phleom unloading     

The Electro-osmotic Theory of Phloem Transport in the Light of Recent Measurements on Heracleum Phloem

The bearing, on the electro-osmotic theory, of work on the Onsagercoefficients for Heracleum phloem, currently reported by Tyreeand Fensom (1970), is briefly discussed. As an aid to the discussionthe Onsager resistance coefficients are evaluated in terms ofSpiegler's frictional model. It is concluded that the experimentalevidence, while it does not support the theory, does not invalidateit. Some further suggestions regarding the electro-osmotic theoryare put forward.     

Phloem loading in squash

Squash (Cucurbita pepo L. var. melopepo torticalis, Bailey) leaves were supplied with ¹⁴C-sucrose, then specific radioactivities of the glucose and galactose moieties of translocated stachyose were determined. In every case, the specific radioactivity of the galactose moiety was greater than that of the glucose moiety. It is concluded that the stachyose was not synthesized at either the phloem-loading site or subsequent to phloem loading, but rather in cells that were not a part of the translocation system, possibly the mesophyll cells.     

Phloem in carrot calluses

Summary Translocation of ¹⁴C-labelled photosynthate across 2 cm long carrot calluses was not detectable even 6 hours after a 30 min ¹⁴CO₂ pulse. This is consistent with the discontinuous nature of the phloem as a whole, although small strands of contiguous, well differentiated sieve cells with companion cells were readily seen in electron micrographs.With the technical collaboration of H. J. W. Edge. Botany Dept. Kings College, London.