Phloem loading strategies and water relations in trees and herbaceous plants

Most herbaceous plants employ thermodynamically active mechanisms of phloem loading, whereas in many trees, the mechanism is passive, by diffusion. Considering the different water transport characteristics of herbs and trees, we hypothesized that water relations play a role in the adoption of phloem loading strategies. We measured whole-plant hydraulic conductance (K(p)), osmolality, concentrations of polar metabolites, and key inorganic ions in recently mature leaves of 45 dicotyledonous species at midafternoon. Trees, and the few herbs that load passively, have low K(p), high osmolality, and high concentrations of transport sugars and total polar metabolites. In contrast, herbs that actively load sucrose alone have high K(p), low osmolality, and low concentrations of sugars and total polar metabolites. Solute levels are higher in sugar alcohol-transporting species, both herbs and trees, allowing them to operate at lower leaf water potentials. Polar metabolites are largely responsible for leaf osmolality above a baseline level (approximately 300 mm) contributed by ions. The results suggest that trees must offset low K(p) with high concentrations of foliar transport sugars, providing the motivating force for sugar diffusion and rendering active phloem loading unnecessary. In contrast, the high K(p) of most herbaceous plants allows them to lower sugar concentrations in leaves. This reduces inventory costs and significantly increases growth potential but necessitates active phloem loading. Viewed from this perspective, the elevation of hydraulic conductance marks a major milestone in the evolution of the herbaceous habit, not only by facilitating water transport but also by maximizing carbon use efficiency and growth.     

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.     

An investigation of bidirectional translocation in the Phloem

Patterns of (14) CO(2) , assimilate movement in Vicia jaba plants having 7 nodes were studied. Bidirectional translocation occurred throughout most of the stem length when tracer was applied to leaves of various ages. To determine whether this bidirectional translocation occurs within single sieve tubes, a O.1 % solution of the fluorescent dye K-fluorescein was applied to a lightly scraped area on the stem in the middle of a young internode. After one hour the dye was present short distances above and below the treated area. Free-hand sections of the internode showed the dye to be localized in the traces of the larger leaves below tbe treated area and in the traces of the younger leaves above the treated area. The dye was never present in the same bundle both above and below the treated area, indicating that each bundle and sieve tube translocated the dye in only one direction. These results were confirmed using Phaseolus vulgaris, Vinca rosea, and Pelargonium hortum. A similar study in which petioles of young Ecballium elaterium leaves were treated showed that usually the phloem of one bundle translocated the dye in only one direction but in some cases the external phloem of the bicollateral bundles carried the dye toward the stem while the internal phloem carried the dye toward the blade. When longer time intervals were used in all these experiments, the dye sometimes appeared in the same phloem areas both above and below the treated area. This is explained by a lateral transfer of tracer within the phloem, either through secondary phloem or through bundle anastomoses at the nodes.     

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.     

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 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.     

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.     

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.     

Phyllotaxy and water relations in tobacco

Summary The relative effectiveness of vascular connections between adjacent leaves of tobacco is demonstrated. It is shown that water movement between adjacent leaves is more difficult than between phyllotactically related leaves. Total and specific resistance ratios between adjacent and phyllotactically related leaves are calculated. These figures indicate that vertical water potential profiles need to be interpreted with a full knowledge of the vascular structure and phyllotaxy as well as gross structural and environmental parameters.     

Seed vigour and water relations in wheat

The laboratory germination (criterion radicle emergence) of seven seed lots of winter wheat cv. Slejpner was similar. However, they differed in vigour as demonstrated by differences in germination after controlled deterioration carried out at a range of seed moisture contents, at two temperatures and for different times. A vigour assessment for each lot was quantified by calculating a value for the seed lot constant K_i, of the viability equation. Germination in lower water potentials reduced the uptake of water, radicle and coleoptile emergence and radicle and coleoptile extension. There was no difference in the water uptake of seed lots differing in vigour. However, seed lots of lower vigour showed less radicle emergence, less coleoptile emergence and shorter radicles than higher vigour seed lots in low water potentials. Similarly, controlled deterioration resulted in reduced radicle and coleoptile emergence and growth compared to unaged seed, and also to a greater sensitivity to low water potentials. The implications for field establishment are discussed.     

Linking water relations and hydraulics with photosynthesis

For land plants, water is the principal governor of growth. Photosynthetic performance is highly dependent on the stable and suitable water status of leaves, which is balanced by the water transport capacity, the water loss rate as well as the water capacitance of the plant. This review discusses the links between leaf water status and photosynthesis, specifically focussing on the coordination of CO₂ and water transport within leaves, and the potential role of leaf capacitance and elasticity on CO₂ and water transport.     

Cytoplasmic gel and water relations of axon

Summary A previous method of measuring the swelling pressure ( _g ) of the cytoplasmic gel of the giant axon ofLoligo vulgaris was refined. The estimates of _g made with the improved method were consistent with those made with the earlier method. In these methods the activity of the solvent in the gel is measured by increasing the activity of the solvent in the internal phase of the gel by application of hydrostatic pressure to the gel directly. Comparable values for the activity of the solvent in the gel were obtained also by an alternate method, in which the deswelling of the gel is measured upon decreasing the activity of the solvent in the external phase by addition of a nonpenetrating high mol wt polymer (i.e., Ficoll).Additional support was obtained for the earlier suggestion that _g contributes to the swelling and shrinkage pattern of the whole axon. In part, the new evidence involved two consecutivedirect measurements of intraxonal pressure. The first measurement was that of a mixed pressure composed of _g and _m ( _m being the effective osmotic pressure due to the intra-extraxonal gradient in the activity of mobile solutes). The subsequent measurement was that of _g alone. The latter measurement was made feasible by destroying the axolemma, thereby eliminating the contribution of _m . An estimate of _m was obtained by subtracting _g from the total pressure measured initially. The _m determined by the above method was two orders of magnitude smaller than the theoretical osmotic pressure. This is consistent with the _m determined previously, where osmotic intra-extraxonal filtration coefficients were compared to the hydrostatic. The mixed pressure experiments lend credence to the idea that the substantial contribution of _g to the water relations of the whole axon is due to _g being of the same order of magnitude as _m .The degree of free swelling of axoplasmic gels was studied as a function of pH, salt concentration, and hydration radius of the anion of the salt used. The swelling increased with an increase in the reciprocal of the hydration radius, a decrease in salt concentration, and at pH below or above 4.5.The nature of the constraints to the free swelling of axoplasm in axons immersed in seawater was studied. With the seawater employed, these constraints appeared to be due more to the retractive forces of the sheath than to _m .     

Temperature and water relations in desert bees

1. 1. Desert bees do not show significant differences in most thermal parameters; mean endothermic warm-up rates are similar to those of temperate species, with no special cooling mechanisms, and normal upper critical temperatures (unlike desert ants and beetles). Thermoregulatory abilities may however be improved.

2. 2. They show the whole range of possible water balance problems; small species are acutely water-stressed when foraging, but large bees suffer from excessive generation of metabolic water in flight.

3. 3. Activity patterns are therefore either matinal, crepuscular or bimodal; essentially desert bees avoid heat and adapt to cold desert dawns and dusks. Desert plants must be coevolved to offer appropriate rewards and match the physiological constraints on their pollinators.

4. 4. Endothermy in bees may have evolved primarily in arid zones, and served as a pre-adaptation for subsequent invasion of cool temperate biomes.