The cellular pathway of photosynthate transfer in the developing wheat grain. I. Delineation of a potential transfer pathway using fluorescent dyes

A potential cellular pathway for photosynthate transfer between the crease phloem and the starchy endosperm of the developing wheat grain has been delineated using fluorescent dyes. Membrane permeable and impermeable dyes have been introduced into the grain through the crease phloem, the endosperm cavity or the dorsal surface of the starchy endosperm. The movement of the symplastic tracer 5-(6)-6-carboxyfluorescein (CF) derived from 5-(6)-6-carboxyfluorescein diacetate (CFDA), from either direction between the crease phloem and the endosperm cavity, indicated that the symplastic pathway was operative from the crease phloem to the nucellar projection. Furthermore, the inward movement of apoplastic tracer trisodium, 3-hydroxy-5,8,10-pyrentrisulphonate (PTS) from the endosperm cavity and that of CF following plasmolysis showed that there was a high resistance to solute transfer within the apoplast of the pigment strand. All dyes entered the modified aleurone and adjacent sub-aleurone bordering the endosperm cavity. Subsequent movement of the symplastic tracers CF and sulphorhodamine G (SRG) into and through the endosperm was rapid. However, the movement of apoplastic tracers PTS and Calcofluor White (CFW) was relatively slow and with tissue plasmolysis, CF was confined to the cytoplasm of the modified aleurone and subaleurone cells. Together, these results demonstrate that there is a high resistance to solute movement within the apoplast of the cells bordering the endosperm cavity. We propose that photosynthate transfer is via the symplast to the nucellar projection where membrane exchange to the endosperm cavity occurs. Uptake from the cavity is by the modified aleurone and small endosperm cells prior to transfer through the symplast to and through the starchy endosperm.     

Phloem loading. A reevaluation of the relationship between plasmodesmatal frequencies and loading strategies

The incidence of plasmodesmata in the minor vein phloem of leaves varies widely between species. On this basis, two pathways of phloem loading have been proposed: symplastic where frequencies are high, and apoplastic where they are low. However, putative symplastic-loading species fall into at least two categories. In one, the plants translocate raffinose-family oligosaccharides (RFOs). In the other, the primary sugar in the phloem sap is sucrose (Suc). While a thermodynamically feasible mechanism of symplastic loading has been postulated for species that transport RFOs, no such mechanism is known for Suc transporters. We used p-chloromercuribenzenesulfonic acid inhibition of apoplastic loading to distinguish between the two pathways in three species that have abundant minor vein plasmodesmata and are therefore putative symplastic loaders. Clethra barbinervis and Liquidambar styraciflua transport Suc, while Catalpa speciosa transports RFOs. The results indicate that, contrary to the hypothesis that all species with abundant minor vein plasmodesmata load symplastically, C. barbinervis and L. styraciflua load from the apoplast. C. speciosa, being an RFO transporter, loads from the symplast, as expected. Data from these three species, and from the literature, also indicate that plants with abundant plasmodesmata in the minor vein phloem have abundant plasmodesmata between mesophyll cells. Thus, plasmodesmatal frequencies in the minor veins may be a reflection of overall frequencies in the lamina and may have limited relevance to phloem loading. We suggest that symplastic loading is restricted to plants that translocate oligosaccharides larger than Suc, such as RFOs, and that other plants, no matter how many plasmodesmata they have in the minor vein phloem, load via the apoplast.     

Sink anatomy in relation to solute movement in rice (Oryza sativa L.): A summary of findings

The results of studies on assimilate and water transport in the developing caryopsis of rice are summarised. Evidence is presented for a symplastic movement of solutes as far as the aleurone layer. However, transport into the apoplast at the nucellus/aleurone interface appears to be a necessary step due to the absence of plasmodesmata at this site. It is suggested that water leaves the caryopsis during grain filling by the isolated cell walls of the pigment strand, the suberised walls of these cells functioning to isolate the apoplast from the symplast and thereby allowing opposing fluxes of water and assimilates to occur in the dorsal region of the grain.     

Nucellar projection transfer cells in the developing wheat grain

Summary Transfer cells in the nucellar projection of wheat grains at 25 ±3 days after anthesis have been examined using light and electron microscopy. Within the nucellar tissue, a sequential increase in non-polarized wall ingrowth differentiation and cytoplasmic density was evident. Cells located near the pigment strand were the least differentiated. The degree of differentiation increased progressively in cells further removed from the pigment strand and the cells bordering the endosperm cavity had degenerated. Four stages of transfer cell development were identified at the light microscope level. Wall ingrowth differentiation followed a sequence from a papillate form through increased branching (antler-shaped ingrowths) which ultimately anastomosed to form a complex labyrinth. The final stage of wall ingrowth differentiation was compression which resulted in massive ingrowths. In parallel with wall ingrowth deposition cytoplasmic density increased. During wall deposition, paramural and multivesicular bodies were prominent and were in close association with the wall ingrowths. The degeneration phase involved infilling of cytoplasmic islets within the wall ingrowths. This was accompanied by complete loss of the protoplast. The significance of this transfer cell development for sucrose efflux to the endosperm cavity was assessed by computing potential sucrose fluxes across the plasma membrane surface areas of the nucellar projection cells. Transfer cell development amplified the total plasma membrane surface area by 22 fold. The potential sucrose flux, when compared with maximal rates of facilitated membrane transport of sugars, indicated spare capacity for sucrose efflux to the endosperm cavity. Indeed, when the total flux was partitioned between the nucellar projection cells at the three stages of transfer cell development, the fully differentiated stage III cells located proximally to the endosperm cavity alone exhibited spare transport capacity. Stage II cells could accommodate the total rate of sucrose transfer, but stage I cells could not. It is concluded that the nucellar projection tissue of wheat provides a unique opportunity to study transfer cell development and the functional role of these cells in supporting sucrose transport.Abbreviations CSPMSA cross sectional plasma membrane surface area - LPMSA longitudinal plasma membrane surface area - PTS tri-sodium 3-hydroxy-5,8,10-pyrenetrisulfonate     

The Use of Fluorescent Tracers to Characterize the Post-Phloem Transport Pathway in Maternal Tissues of Developing Wheat Grains

Various polar fluorescent tracers were used to characterize the pathways for apoplastic and symplastic transport in the "crease tissues" (i.e. the vascular strand, chalaza, nucellus, and adjacent pericarp) of developing wheat (Triticum aestivum L.) grains. With mostly minor exceptions, the results strongly support existing views of phloem unloading and post-phloem transport pathways in the crease. Apoplastic movement of Lucifer yellow CH (LYCH) from the endosperm cavity into the crease was virtually blocked in the chalazal cell walls before reaching the vascular tissue. However, LYCH could move slowly along the cell wall pathway from the chalaza into the vascular parenchyma. Slow uptake of LYCH into nucellar cell cytoplasm was observed, but no subsequent symplastic movement occurred. Carboxyfluorescein (CF) imported into attached grains moved symplastically from the phloem across the chalaza and into the nucellus, but was not released from the nucellus. In addition, CF moved in the opposite direction (nucellus to vascular parenchyma) in attached grains. Thus, the post-phloem symplastic pathway can accommodate bidirectional transport even when there is an intense net assimilate flux in one direction. When fresh sections of the crease were placed in fluorochrome solutions (e.g. LYCH or pyrene trisulfonate), dye was rapidly absorbed into intact cells, apparently via unsealed plasmodesmata. Uptake was not visibly reduced by cold or by respiratory inhibitors, but was greatly reduced by plasmolysis. Once absorbed, the dye moved intercellularly via the symplast. Based on this finding, a size-graded series of fluorescein-labeled dextrans was used to estimate the size-exclusion limits (SEL) for the post-phloem symplastic pathway. In most, and perhaps all, cells of the crease tissues except for the pericarp, the molecular diameter for the SEL was about 6.2 nm. The SEL in much of the vascular parenchyma may be smaller, but it is still at least 3.6 nm. Channel diameters would likely be about 1 nm larger, or about 4.5 to 7.0 nm in the vascular parenchyma and 7.0 nm elsewhere. These dimensions are substantially larger than those for "conventional" symplastic connections (about 3 nm), and would have a greater than proportionate effect on the per channel diffusive and hydraulic conductivities of the pathway. Thus, relatively small and probably ultrastructurally undetectable adjustments in plasmodesmatal structure may be sufficient to account for assimilate flux through the crease symplast.     

The cellular pathway of photosynthate transfer in the developing wheat grain. III. A structural analysis and physiological studies of the pathway from the endosperm cavity to the starchy endosperm

The cellular pathway of sucrose transfer from the endosperm cavity to the starchy endosperm of developing grains of wheat (Triticum turgidum) has been elucidated. The modified aleurone and sub-aleurone cells exhibit a dense cytoplasm enriched in mitochondria and endoplasmic relicilium. Significantly, the sub-aleurone cells are characterized by secondary wall ingrowths. Numerous plasmodesmata interconnect all cells between the modified aleurone and starchy endosperm. The pro-tonophore carbonylcyanide-m-chlorophenyl hydrazone (CCCP) slowed [¹⁴C]sucrose uptake by grain tissue slices enriched in modified aleurone and sub-aleurone cells but had no effect on uptake by the starchy endosperm. The fluorescent weak acid sulphorhodamine G (SRG) was preferentially accumulated by the modified aleurone and sub-aleurone cells, and this uptake was sensitive to CCCP. The combined plasma membrane surface areas of the modified aleurone and sub-aleurone cells appeared to be sufficient to support the in vivo rates of sucrose transfer to the starchy endosperm. Plasmolysis of intact excised grain inhibited [¹⁴C]sucrose transfer from the endosperm cavity to the starchy endosperm. The sulphydryl group modifier p-chloromercuribenzenesulphonie acid (PCMBS) decreased [¹⁴C]sucrose uptake by the modified aleurone and sub-aleurone cells but had little effect on uptake by the starchy endosperm. In contrast, when PCMBS and [¹⁴C]sucrose were supplied to the endosperm cavity of intact excised grain, PCMBS slowed accumulation by all tissues equally. Estimates of potential sucrose fluxes through the interconnecting plasmodesmata were found to be within the published range. It is concluded that the bulk of sucrose is accumulated from the endosperm cavity by the modified aleurone and sub-aleurone cells and subsequently transferred through the symplast to the starchy endosperm.     

Pathway of Photosynthate Transfer in the Developing Seed of Vicia faba L: A Structural Assessment of the Role of Transfer Cells in Unloading from the Seed Coat

Photosynthate movement within the coat of the developing seedof Vicia faba occurs radially inward from the restricted vascularsystem and laterally through the non-vascularized region ofthe seed coat prior to exchange to the seed apoplast. Thin-walledparenchyma/transfer cells line the entire inner surface of theseed coat and thus are located at the terminus of the photosynthatetransfer pathway. The principal cellular route of transfer withinthe seed coat and the role of the thin-walled parenchyma/transfercells in membrane exchange to the seed apoplast has been investigated.Sucrose fluxes, computed from estimates of the plasma membranesurface areas of the cell types of the pathway, the plasmodesmatalcross-sectional areas interconnecting contiguous cells and theobserved rate of sucrose delivery to the embryo indicate thatsieve element unloading and subsequent transfer to the thin-walledparenchyma/transfer cells is through the symplast. For the cellsof the ground tissue, plasmodesmatal density is consistentlyhigher on their anticlinal walls. This observation supportsthe reported pattern of lateral transfer through these tissuesin the non-vascular regions of the seed coat. Wall ingrowthsare initiated sequentially in the thin-walled parenchyma cellsto maintain 1–3 rows of thin-walled parenchyma/transfercells. The development of these wall ingrowths results in a58% increase in the plasma membrane surface area of these cellsand provides them with the capacity to act as the principalcellular site for membrane exchange of sucrose to the seed apoplast.This cellular route of symplastic transfer from the sieve elementsto the ground tissues where membrane exchange to the seed apoplastoccurs is consistent with that reported for Phaseolus vulgaris Key words: Cellular pathway, photosynthate transfer, transfer cell, Vicia seed coat     

Transport of assimilates in the developing caryopsis of rice (Oryza sativa L.)

Assimilates entering the developing rice caryopsis traverse a short-distance pathway between the terminal sieve elements of the pericarp vascular bundle and the aleurone layer. The ultrastructure of this pathway has been studied. Sieve elements in the pericarp vascular bundle are smaller than their companion cells.The sieve elements show few connections with surrounding vascular parenchyma elements but are connected to companion cells by compound plasmodesmata. Companion cells, in turn, are connected to vascular parenchyma elements by numerous compound plasmodesmata present in wall thickenings. Assimilates leaving the sieve element — companion cell complex must laterally traverse cells of the pigment strand before they come into contact with the aleurone layer. The pigment strand cells have modified inner walls made up of a suberin-like material. This material may act as a permeability barrier isolating the apoplast from the symplast of the pigment strand. The walls of the pigment strand cells are traversed by numerous plasmodesmata. Water may be conducted to the endosperm through the isolated cell-wall system of the pigment strand while assimilates possibly move via plasmodesmata. High frequencies of plasmodesmata occur at the junction between the pigment strand and the nucellus and also between adjacent cells of the nucellus. By contrast, plasmodesmata are absent between the nucellus and the aleurone layer and also between the nucellus and the seed coat. A predominantly circumferential and symplastic transport pathway is likely between the pigment strand and nucellus. In view of the total absence of plasmodesmata between the nucellus and the aleurone layer assimilates entering the endosperm may have to cross the plasmalemma of the nucellus. It is possible that constraints to the flow of assimilates may occur in the short-distance pathway between the terminal sieve element — companion cell complexes and the endosperm, and this is discussed.     

Sieve element unloading: cellular pathway, mechanism and control

The transport and distribution of phloem – mobile solutes is predominantly determined by transport processes located at the sink end of the source – transport – sink system. Transport across the sieve element boundary, sieve element unloading, is the first of a series of sink transport processes. Unloading of solutes from the sieve elements may follow an apo- or symplastic route. It is speculated that the unloading pathway is integrated with sink function and that apoplastic unloading is restricted to situations in which movement through the symplast is not compatible with sink function. These situations include axial transport and storage of osmotically active solutes against concentration and turgor gradients between the sieve elements and sink cells. Coupled with alteration in sink function, the cellular pathway of unloading can switch in stems and possibly other sinks. Experimental systems and approaches used to elucidate the mechanism of sieve element unloading are reviewed. Unloading fluxes to the apoplast can largely be accounted for by membrane diffusion in axial sinks. However, the higher fluxes in storage sinks suggests dependence on some form of facilitated transport. Proton sucrose symport is assessed to be a possible mechanism for facilitated efflux of solutes across the sieve element plasma membrane to the sink apoplast. Unloading through the symplast may occur by diffusion or mass flow. The latter mechanism serves to dissipate phloem water and hence prevent the potential elevation of sieve element turgor that would otherwise slow phloem import into the sink. The possibility of energised plasmodesmatal transport is raised. Sieve element unloading must be integrated with subsequent compartmentation and metabolism of the unloaded solute. Solute levels are an obvious basis for control of sieve element unloading, but are found to offer limited scope for a mass action mechanism. Apoplastic, cellular pathway, sieve element, solute transport, symplastic. Translated into a turgor signal, solute levels could regulate the rate of unloading, metabolism and compartmentation forming part of a turgor homeostat irrespective of the pathway of unloading.     

The Cellular Pathway of Short-distance Transfer of Photosynthates and Potassium in the Elongating Stem of Phaseolus vulgaris L. A Structural Assessment

The potential cellular pathway of radial transfer of photosynthateand potassium delivered in the phloem to the elongation zone(apical 0.5–2.5 cm) of internode 2 ofPhaseolus vulgarisL. seedlings was elucidated. This was achieved using ultrastructuralobservations of the cell types that constitute the radial pathwayand estimates of potential sucrose and potassium fluxes throughthe cross-sectional area of interconnecting plasmodesmata andacross the plasma membrane surface areas of selected cell types.The investigation relied on predicting the relative roles ofthe mature and developing sieve elements as conduits for theaxial delivery of solutes to the elongation zone. In turn, thesepredictions led to formulation of two transport models whichwere subsequently evaluated. It was found that unloading ofsucrose and potassium from the protophloem sieve elements cannotbe through the symplast due to the absence of plasmodesmata.On the other hand, mature metaphloem sieve element-companioncell complexes have the potential capacity to unload eitherthrough the stem symplast or apoplast. The potential symplasticroute is proposed to be via the companion cells to the adjacentlarge phloem parenchyma cells. Continued radial transfer couldoccur either by exchange to the stem apoplast from the largephloem parenchyma cells or continue in the symplast to the groundtissues. It was further predicted that sucrose utilized forthe development of the procambial/small phloem parenchyma cellscould be delivered axially by them and not by the mature sieveelements. Phaseolus vulgaris ; apoplast; elongating stem; photosynthates; potassium; transport; symplast     

Cellular Structures, Plasma Membrane Surface Areas and Plasmodesmatal Frequencies of the Stem of Phaseolus vulgaris L. in Relation to Radial Photosynthate Transfer

At an early stage of secondary development, the metaphloem sieveelements appeared to be the only functional axial transportconduit in fully elongated stems of P. vulgaris plants. Thereis no apparent barrier to the radial transfer of solutes inthe stem apoplast. However, radial transfer through the stemsymplast could be limited by discontinuities resulting fromprotoplast degeneration of the protophloem fibres and developingsecondary xylem fibres. Estimates of possible sucrose fluxesthrough the apoplastic and symplastic routes indicated thatradial photosynthate transfer from the sieve element-companioncell (se-cc) complexes of the stem metaphloem could follow eithercellular route. In the case of apoplastic transfer, the plasmamembrane surface area of the se-cc complexes is only sufficientto support some form of facilitated movement of sucrose. Incontrast, the plasma membrane surface area of the phloem parenchymais sufficient to permit passive diffusion of sucrose to theapoplast. Plasmodesmatal frequencies suggest that any symplastictransfer to the phloem parenchyma from the sieve elements wouldbe via the companion cells. Phaseolus vulgaris, french bean, stem, photosynthate, radial transfer (photosynthates), cellular pathway     

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.     

Water Flow Across the Sieve-Tube Boundary: Estimating Turgor and Some Implications For Phloem Loading and Unloading. I. Theory

A mathematical model of water and sucrose transport across thesieve tube boundary is presented, based on conservation of matterand the phenomenological equations for plasmodesmatal transportbetween the sieve elements and their associated cells. Plasmodesmataltransport coefficients are discussed. In parts II–IV,the equations developed here are used to assess: (i) the estimationof phloem turgor gradients from osmotic pressure gradients;(ii) plasmodesmatal transport of water and sucrose between thesieve elements and adjacent cells; and (iii) the plausibilityof symplastic and apoplastic phloem loading and unloading insome primary sources and sinks. A list of symbols is given inAppendix 1 of this paper Phloem, turgor, osmotic pressure, loading, unloading, plasmodesmata, Munch hypothesis     

Compartmentation of Assimilate Fluxes in Leaves

Abstract: Sugar levels in the apoplast of assimilate exporting leaves were studied in two groups of plant species with contrasting structures of companion cells in minor veins. These species are termed either "symplastic" (with intermediary cells) or "apoplastic" (with transfer or ordinary cells). Sugars were measured in intercellular washing fluid after extracting the apoplast by an infiltration-centrifugation technique. During the course of a day, sugar contents in the apoplast were, in general, lower in species with intermediary cells than in species with transfer or ordinary cells. In "symplastic" species, apoplastic sucrose concentrations were between 0.3 and 1 mM. In "apoplastic" species with transfer cells, they ranged between 2 and 6 mM. Apoplastic hexose contents were between 0.3 and 1 mM irrespective of presumed transport mode. "Symplastic" and "apoplastic" plants differed markedly in their response to a'translocation block. In "symplastic" plants, inhibition of assimilate export left apoplastic concentrations of sucrose and hexoses unchanged, whereas in "apoplastic" plants sugar levels increased, the maximal increase being observed with sucrose. In these plants, concentrations of sucrose were two to six times higher in the apoplast under export inhibition than in control leaves. The data suggest a different role of the leaf apoplast in the compartmentation and export of assimilates in the two plant groups under study.     

灵武长枣果实ATPase超微细胞化学定位和功能研究

采用ATPase超微细胞化学定位技术,研究灵武长枣果实不同发育阶段韧皮部和果肉库薄壁细胞ATPase分布特征,以明确灵武长枣果实ATPase超微细胞化学定位特征和功能。结果显示:(1)第一次快速生长期SE/CC复合体与周围的薄壁细胞有丰富的胞间连丝,形成共质体连续,韧皮部薄壁细胞之间有丰富的胞间连丝,ATPase反应物在韧皮部各细胞分布较少。(2)缓慢生长期ATPase反应物在韧皮部各细胞分布逐渐增加。(3)第二次快速生长期SE/CC复合体与周围的薄壁细胞缺乏胞间连丝,形成共质体隔离,韧皮薄壁细胞及果肉库薄壁细胞的胞间连丝较少,囊泡和膜泡在筛管、韧皮薄壁细胞和库薄壁细胞中很丰富,质膜、液泡膜、囊泡膜、细胞壁和胞间隙的ATPase活性较高。研究表明,果实在第一次快速生长期同化物从筛分子的卸出主要采取共质体途径,缓慢生长期同化物卸出时可能为共质体和质外体途径共存,第二次快速生长期则主要以质外体途径为主,证明果实不同发育阶段韧皮部同化物卸出路径存在差异。     

Movement of fluorescein into isolated caryopses of wheat and barley

Abstract. The movement of fluorescein, a symplastic fluorescent tracer, into isolated caryopses of wheat and barley is described. The dye followed the pathway to the endosperm which has been proposed previously from anatomical studies, namely a movement from the phloem, through cells of the pigment strand and nucellar projection, followed by a radial spread of the dye from the endosperm cavity into the starchy endosperm. By contrast, the fluorochromes calcofluor white M2R and ANS remained confined to the apoplast and failed to cross the 'xylem discontinuity' at the base of the caryopses.     

<Emphasis Type="Italic">Amborella trichopoda</Emphasis>, plasmodesmata,and the evolution of phloem loading

Phloem loading is the process by which photoassimilates synthesized in the mesophyll cells of leaves enter the sieve elements and companion cells of minor veins in preparation for long distance transport to sink organs. Three loading strategies have been described: active loading from the apoplast, passive loading via the symplast, and passive symplastic transfer followed by polymer trapping of raffinose and stachyose. We studied phloem loading in Amborella trichopoda, a premontane shrub that may be sister to all other flowering plants. The minor veins of A. trichopoda contain intermediary cells, indicative of the polymer trap mechanism, forming an arc on the abaxial side and subtending a cluster of ordinary companion cells in the interior of the veins. Intermediary cells are linked to bundle sheath cells by highly abundant plasmodesmata whereas ordinary companion cells have few plasmodesmata, characteristic of phloem that loads from the apoplast. Intermediary cells, ordinary companion cells, and sieve elements form symplastically connected complexes. Leaves provided with ¹⁴CO₂ translocate radiolabeled sucrose, raffinose, and stachyose. Therefore, structural and physiological evidence suggests that both apoplastic and polymer trapping mechanisms of phloem loading operate in A. trichopoda. The evolution of phloem loading strategies is complex and may be difficult to resolve.     

What Is Phloem Unloading?

Several studies of phloem unloading have failed to distinguish between transport events occurring at the sieve element/companion cell boundary and subsequent short-distance transport through parenchyma cells. Indirect evidence has been obtained for symplastic unloading in storage and utilization sinks. In other sinks transfer to the apoplast may occur, but not necessarily at the sieve element/companion cell complex, and the evidence for apoplastic phloem unloading is equivocal, as is the role of apoplastic acid invertase in this process. The ability of several types of sink cells to accumulate sugars from the apoplast is discussed in the conflicting light of functional symplastic continuity between sink cells. Attention is drawn to the complexity of the postunloading pathway in many sinks and the difficulty of determining the exact sites of symplast/apoplast solute exchange. Potential future areas for study in the field are highlighted.     

灵武长枣果实维管束韧皮部及周围细胞的超微结构特征

为了探讨灵武长枣果实光合同化物韧皮部卸载和运输的途径,该研究采用透射电镜技术,对不同发育时期灵武长枣果实维管束韧皮部及其周围薄壁细胞的超微结构特征进行了分析。结果表明:筛管/伴胞复合体及其周围韧皮薄壁细胞间在果实膨大前期富含胞间连丝,而韧皮薄壁细胞与周围库细胞以及相邻库细胞间几乎不存在胞间连丝,形成共质体隔离;筛管/伴胞复合体及其与周围薄壁细胞间在果实快速膨大期也存在胞间连丝,但与果实膨大前期相比明显减少;果实着色期,SE/CC复合体及其与周围薄壁细胞间胞间连丝较少,并且出现阻塞现象;果实完熟期,筛管和伴胞之间几乎没有胞间连丝,有的筛管之间有少量胞间连丝,但却出现了阻塞现象,果肉库薄壁细胞与韧皮薄壁细胞间因胞间连丝阻塞现象而形成共质体隔离。综上结果认为,在果实发育的膨大前期阶段,光合同化物以共质体途径经筛分子卸出,卸出后可能以质外体途径进入液泡贮藏与利用;果实快速膨大期,光合同化物的卸出与运输采用共质体和质外体共存的途径;果实着色期和完熟期,光合同化物从筛分子卸出到贮藏薄壁细胞的运输均以质外体途径为主。     

Phloem unloading in the potato tuber. Pathways and sites of ATPase

Summary Potential pathways for sucrose unloading in the potato tuber were examined by light and electron microscopy. Abundant plasmodesmata connected sieve elements with surrounding parenchyma elements and also sieve elements with companion cells. Plasmodesmata were rarer, however, between companion cells and parenchyma elements. These observations suggest that sucrose may leave the sieve elements and enter the storage parenchyma cells directly via the symplast and that transport through the companion cell may not be a prerequisite for unloading. Plasmodesmata, grouped together in primary pit fields, were also abundant between storage cells, and isolated storage cells, separated enzymically, showed considerable variation in plasmodesmatal distribution between cells and also on different faces of a single cell. Deposition of starch was found to occur in the tuber cortex while an endodermis with Casparian strip was present external to the phloem, suggesting that assimilates initially enter the cortical storage cells by an entirely symplastic pathway. The possible involvement of ATPase in the unloading process was examined cytochemically, using a lead-salt precipitation method. By contrast with previous findings for phloem no evidence was found for ATPase activity that was unique to the sieve element-companion cell complex. The present observations favour the view that phloem unloading in the potato tuber is a symplastic and passive process.