Sucrose Flux Research Articles

Cell Wall Invertases from Maternal Tissues Modulate Sucrose Flux in Apoplastic Pathways During Rice Anther and Seed Development

Efficient sucrose transport and metabolism are vital for seed and pollen development in plants. Cell wall invertases (CINs) hydrolyze sucrose into glucose and fructose, maintaining a sucrose gradient in the apoplast of sink tissues. In rice, two CIN isoforms, OsCIN1 and OsCIN2, were identified as being specifically expressed in the anthers but not in pollen. Functional analyses through genetic crosses and mutant characterization showed that oscin1/2 double mutants exhibit a sporophytic male-sterile phenotype and produce shrunken seeds. This suggests that CIN activity is essential for proper pollen development and seed formation in rice. Observation of the progeny genotypes and phenotypes from various genetic crosses revealed that the phenotype of oscin1/2 seeds is determined by the genotype of the maternal tissue, indicating the critical role of CIN function in the apoplast between maternal and filial tissues for sucrose transport and metabolism. The CIN activity in the anthers and seeds of wild-type rice was found to be significantly higher—over 500-fold in the anthers and 5-fold in the seeds—than in the leaves, highlighting the importance of CIN in facilitating the efficient unloading of sucrose. These findings suggest that the fine-tuning of CIN activity in the apoplast, achieved through tissue-specific expression and CIN isoform regulation, plays a key role in determining the carbohydrate distribution across different tissues. Understanding this regulatory mechanism could provide opportunities to manipulate carbohydrate allocation to sink organs, potentially enhancing crop yields.

Transcriptomic analysis of mesocarp tissue during fruit development of the oil palm revealed specific isozymes related to starch metabolism that control oil yield.

The oil palm (Elaeis guineensis Jacq.) produces a large amount of oil from the fruit. However, increasing the oil production in this fruit is still challenging. A recent study has shown that starch metabolism is essential for oil synthesis in fruit-producing species. Therefore, the transcriptomic analysis by RNA-seq was performed to observe gene expression alteration related to starch metabolism genes throughout the maturity stages of oil palm fruit with different oil yields. Gene expression profiles were examined with three different oil yields group (low, medium, and high) at six fruit development phases (4, 8, 12, 16, 20, and 22 weeks after pollination). We successfully identified and analyzed differentially expressed genes in oil palm mesocarps during development. The results showed that the transcriptome profile for each developmental phase was unique. Sucrose flux to the mesocarp tissue, rapid starch turnover, and high glycolytic activity have been identified as critical factors for oil production in oil palms. For starch metabolism and the glycolytic pathway, we identified specific gene expressions of enzyme isoforms (isozymes) that correlated with oil production, which may determine the oil content. This study provides valuable information for creating new high-oil-yielding palm varieties via breeding programs or genome editing approaches.

Auxin boosts energy generation pathways to fuel pollen maturation in barley

Pollen grains become increasingly independent of the mother plant as they reach maturity through poorly understood developmental programs. We report that the hormone auxin is essential during barley pollen maturation to boost the expression of genes encoding almost every step of heterotrophic energy production pathways. Accordingly, auxin is necessary for the flux of sucrose and hexoses into glycolysis and to increase the levels of pyruvate and two tricarboxylic (TCA) cycle metabolites (citrate and succinate). Moreover, bioactive auxin is synthesized by the pollen-localized enzyme HvYUCCA4, supporting that pollen grains autonomously produce auxin to stimulate a specific cellular output, energy generation, that fuels maturation processes such as starch accumulation. Our results demonstrate that auxin can shift central carbon metabolism to drive plant cell development, which suggests a direct mechanism for auxin's ability to promote growth and differentiation.

Rubisco regulation in response to altered carbon status in the cyanobacterium Synechococcus elongatus PCC 7942.

Photosynthetic organisms possess a variety of mechanisms to achieve balance between absorbed light (source) and the capacity to metabolically utilize or dissipate this energy (sink). While regulatory processes that detect changes in metabolic status/balance are relatively well studied in plants, analogous pathways remain poorly characterized in photosynthetic microbes. Here, we explored systemic changes that result from alterations in carbon availability in the model cyanobacterium Synechococcus elongatus PCC 7942 by taking advantage of an engineered strain where influx/efflux of a central carbon metabolite, sucrose, can be regulated experimentally. We observed that induction of a high-flux sucrose export pathway leads to depletion of internal carbon storage pools (glycogen) and concurrent increases in estimates of photosynthetic activity. Further, a proteome-wide analysis and fluorescence reporter-based analysis revealed that upregulated factors following the activation of the metabolic sink are concentrated on ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) and auxiliary modules involved in Rubisco maturation. Carboxysome number and Rubisco activity also increased following engagement of sucrose secretion. Conversely, reversing the flux of sucrose by feeding exogenous sucrose through the heterologous transporter resulted in increased glycogen pools, decreased Rubisco abundance, and carboxysome reorganization. Our data suggest that Rubisco activity and organization are key variables connected to regulatory pathways involved in metabolic balancing in cyanobacteria.

Concurrent overexpression of amino acid permease

Using pea as our model crop, we sought to understand the regulatory control over the import of sugars and amino acids into the developing seeds and its importance for seed yield and quality. Transgenic peas simultaneously overexpressing a sucrose transporter and an amino acid transporter were developed. Pod walls, seed coats, and cotyledons were analysed separately, as well as leaves subtending developing pods. Sucrose, starch, protein, free amino acids, and endogenous cytokinins were measured during development. Temporal gene expression analyses (RT-qPCR) of amino acid (AAP), sucrose (SUT), and SWEET transporter family members, and those from cell wall invertase, cytokinin biosynthetic (IPT) and degradation (CKX) gene families indicated a strong effect of the transgenes on gene expression. In seed coats of the double transgenics, increased content and prolonged presence of cytokinin was particularly noticeable. The transgenes effectively promoted transition of young sink leaves into source leaves. We suggest the increased flux of sucrose and amino acids from source to sink, along with increased interaction between cytokinin and cell wall invertase in developing seed coats led to enhanced sink activity, resulting in higher cotyledon sucrose at process pea harvest, and increased seed number and protein content at maturity.

The Role of Rice Vacuolar Invertase2 in Seed Size Control.

Sink strength optimizes sucrose import, which is fundamental to support developing seed grains and increase crop yields, including those of rice (Oryza sativa). In this regard, little is known about the function of vacuolar invertase (VIN) in controlling sink strength and thereby seed size. Here, in rice we analyzed mutants of two VINs, OsVIN1 and OsVIN2, to examine their role during seed development. In a phenotypic analysis of the T-DNA insertion mutants, only the OsVIN2 mutant osvin2-1 exhibited reduced seed size and grain weight. Scanning electron microscopy analysis revealed that the small seed grains of osvin2-1 can be attributed to a reduction in spikelet size. A significant decrease in VIN activity and hexose level in the osvin2-1 spikelets interfered with spikelet growth. In addition, significant reduction in starch and increase in sucrose, which are characteristic features of reduced turnover and flux of sucrose due to impaired sink strength, were evident in the pre-storage stage of osvin2-1 developing grains. In situ hybridization analysis found that expression of OsVIN2 was predominant in the endocarp of developing grains. A genetically complemented line with a native genomic clone of OsVIN2 rescued reduced VIN activity and seed size. Two additional mutants, osvin2-2 and osvin2-3 generated by the CRISPR/Cas9 method, exhibited phenotypes similar to those of osvin2-1 in spikelet and seed size, VIN activity, and sugar metabolites. These results clearly demonstrate an important role of OsVIN2 as sink strength modulator that is critical for the maintenance of sucrose flux into developing seed grains.

Diurnal dynamics of phloem loading: theoretical consequences for transport efficiency and flow characteristics.

Phloem transport is the process by which plants internally distribute assimilates. The loading of assimilates near the photosynthetic source is responsible for generating enough osmotic pressure to drive sap flow towards the sink tissues where assimilates are consumed. Phloem loading is variable and subject to a diurnal cycle. It is dominated by photosynthesis during the day and by degradation of leaf starch to sugars at night. Most studies ignore the effect of the loading cycle on transport and assume that sugar flow operates at equilibrium. In this study, phloem transport was simulated for three successive days using a finite element model of time-dependent Münch-Horwitz equations. The spatial and temporal distributions of phloem pressure, sucrose concentration, sap velocity and sucrose flux were predicted for five different time variations in sucrose loading. Results showed that periodic loading induces an alternance of two distinct transport phases: one with high pressure, concentration and sucrose flux magnitudes and another with low magnitudes. In contrast, phloem water velocity remained remarkably stable. The alternating phases persisted over time and, under source-driven variation, transport did not reach steady-state conditions for the tested configuration. However, the impact of loading dynamics on transport was mitigated by pathway effects. Oscillations were not only delayed as one travelled away from the source, their amplitude was also reduced over distance. That behaviour stabilized the supply of sucrose to the sink, which continued at moderate levels during the dark cycles. This finding suggests that transport would assist night conversion of starch to sugars in the leaf to prevent carbon starvation at distant sinks in the early morning. The propagation velocity of pressure/concentration waves in phloem was predicted to vary by a factor up to 2.5 depending on the time series chosen to describe the dynamics of loading. Finally, the model predicted that up to 87% of the amount of sucrose loaded over 48 h would be unloaded under time-dependent loading, whereas only 76% would under constant-rate loading. This additional efficiency was periodic. It did not increase significantly the overall efficiency of the system but could be responsible for inducing rhythms in sink activity.

Sugars en route to the roots. Transport, metabolism and storage within plant roots and towards microorganisms of the rhizosphere.

In plants, the root is a typical sink organ that relies exclusively on the import of sugar from the aerial parts. Sucrose is delivered by the phloem to the most distant root tips and, en route to the tip, is used by the different root tissues for metabolism and storage. Besides, a certain portion of this carbon is exuded in the rhizosphere, supplied to beneficial microorganisms and diverted by parasitic microbes. The transport of sugars toward these numerous sinks either occurs symplastically through cell connections (plasmodesmata) or is apoplastically mediated through membrane transporters (MST, mononsaccharide tranporters, SUT/SUC, H+/sucrose transporters and SWEET, Sugar will eventually be exported transporters) that control monosaccharide and sucrose fluxes. Here, we review recent progresses on carbon partitioning within and outside roots, discussing membrane transporters involved in plant responses to biotic and abiotic factors.

Transport in a coordinated soil-root-xylem-phloem leaf system

Abstract Links between the carbon and water economies of plants are coupled by combining the biochemical demand for atmospheric CO2 with gas transfer through stomates, liquid water transport in the soil-xylem hydraulic system and sucrose export in the phloem. We formulated a model to predict stomatal conductance (gs), consistent with the maximum energy circulation concept of Lotka and Odum, by maximizing the sucrose flux out of photosynthesizing leaves. The proposed modeling approach recovers all prior results derived from stomatal optimization theories and profit-maximization arguments for the xylem hydraulic system aimed at predicting gs. The novel features of this approach are its ability to 1) predict the price of losing water in carbon units using xylem and phloem properties (i.e., the marginal water use efficiency) and 2) explain why water molecules become more expensive to exchange for CO2 molecules when soil moisture becomes limiting or when plants acclimate to new elevated atmospheric CO2 concentration. On short time-scales (sub-daily), predicted gs under many environmental stimuli were consistent with measurements reported in the literature, including a general sensitivity of gs to vapor pressure deficit and leaf water potential. During progressive droughts, differences in the coordination among the leaf, xylem, and phloem functioning determine the isohydric-to-anisohydric behavior among plants.

A Gene-Oriented Haplotype Comparison Reveals Recently Selected Genomic Regions in Temperate and Tropical Maize Germplasm.

The extensive genetic variation present in maize (Zea mays) germplasm makes it possible to detect signatures of positive artificial selection that occurred during temperate and tropical maize improvement. Here we report an analysis of 532,815 polymorphisms from a maize association panel consisting of 368 diverse temperate and tropical inbred lines. We developed a gene-oriented approach adapting exonic polymorphisms to identify recently selected alleles by comparing haplotypes across the maize genome. This analysis revealed evidence of selection for more than 1100 genomic regions during recent improvement, and included regulatory genes and key genes with visible mutant phenotypes. We find that selected candidate target genes in temperate maize are enriched in biosynthetic processes, and further examination of these candidates highlights two cases, sucrose flux and oil storage, in which multiple genes in a common pathway can be cooperatively selected. Finally, based on available parallel gene expression data, we hypothesize that some genes were selected for regulatory variations, resulting in altered gene expression.

Effect of water and resource availability on ant feeding preferences: a field experiment on the Mediterranean ant Crematogaster scutellaris

Ants’ feeding habits are affected by individual and collective needs, which may vary among seasons. In this study, we tested the food preferences of the Mediterranean ant Crematogaster scutellaris toward sucrose and amino acid solutions at increasing concentrations, during spring and summer, by analyzing individual choices and mass recruitment in the field. Given that water may be limiting in summer, we also analyzed the role of water as a resource itself. Finally, to investigate how previous short-term availability of resources affects feeding choices, we over-supplied colonies with a continuous flux of amino acids, sucrose and water, before conducting individual tests. As for sucrose, only the most concentrated solutions were largely accepted during the spring, whereas all the solutions were equally taken during the summer. On the contrary, the average acceptance of all amino acid solutions was high only during summer and low in spring. Similar results emerged for recruitments on both nutrients. Amino acid supplementation had no effect on resource acceptance, whereas both sucrose and water supplementation affected the acceptance of all other resources. This study provides hints on the factors affecting seasonal variations in the uptake of carbohydrates and amino acids, and more importantly, clearly shows how food choice is affected by water availability, a factor frequently overlooked in the study of nutritional ecology of ants.

The Role of Temperature on the Diurnal Sucrose Source to Sink Balance

The diurnal fluctuations of carbon from its assimilation within the leaves, its export into the phloem, and its use in sink tissues has been studied in its individual parts for quite some time. In cotton (Gossypium hirsutum L.), the majority of the research concerning diurnal sucrose flux in response to temperature is for the most part greater than a quarter century in age, well before the introduction of modern cultivars. Additionally, diurnal research is lacking of any significant examination of any particular temperature profiles, making it difficult to compare responses from one treatment to another. This summary of research focuses on the diurnal flux occurring from both the source and the sink with special attention on four specific temperature profiles; low night temperatures, high night temperatures, low day temperatures, and high day temperatures. Due to the lack of suitable information inherent in some areas, information from other species was used as needed. It is the authors’ intent that temperature dependent diurnal flux should be reexamined as modern cultivars continue to be developed.

Mathematical modelling of diurnal regulation of carbohydrate allocation by osmo-related processes in plants.

Plants synthesize sucrose in source tissues (mainly mature leafs) and supply it for growth of sink tissues (young leafs). Sucrose is derived from photosynthesis during daytime and from starch at night. Because the diurnal regulation of sucrose fluxes is not completely understood, we built a mathematical model designed to reproduce all key experimental observations. For this, assumptions were made about the molecular mechanisms underlying the regulations, which are all motivated by experimental facts. The key regulators in our model are two kinases (SnRK1 and osmo-sensitive kinase OsmK) under the control of the circadian clock. SnRK1 is activated in the night to prepare for regularly occurring carbon-limiting conditions, whereas OsmK is activated during the day to prepare for water deficit, which often occurs in the afternoon. Decrease of SnRK1 and increase of OsmK result in partitioning of carbon towards sucrose to supply growing sink tissues. Concomitantly, increasing levels of the growth regulator trehalose-6-phosphate stimulates the development of new sink tissues and thus sink demand, which further activates sucrose supply in a positive feedback loop. We propose that OsmK acts as a timer to measure the length of the photoperiod and suggest experiments how this hypothesis can be validated.

Flux control analysis of a lactate and sucrose metabolic network at different storage temperatures for Hami melon (Cucumis melo var. saccharinus)

Cold storage of fruits and vegetables is essentially a respiring and metabolic network flow and control process for which the further study of metabolic control rules are needed in order to improve the shelf-life of these living food products. This study used Hami melon as a test subject to study the respiratory metabolic network and flux control of fruits and vegetables during their storage. Hami melon was stored at different temperatures (0, 5, 10, 15, 20, 25, 30, 35, 40, 45°C), in order to obtain the metabolic flux of the following pathways: Embden–Meyerh of pathway (EMP), pentose phosphate pathway (PPP), sucrose synthesis pathway (SSP) and TCA cycle (TCA). The quantity and activities of primary enzymes (enzymatic expression variate) were measured during storage. In addition, we got the control coefficients of synthesis flux to sucrose and metabolic flux to lactate. The results showed that sucrose had a higher synthesis flux at the temperatures of 5°C and 45°C, and lactate had a higher metabolic flux at the temperature of 5°C. The study on flux control coefficients indicated that sucrose phosphate synthase (SPS, FCC=0.424) produced a larger impact on sucrose flux control in the sucrose synthesis pathway. Lactic dehydrogenase (LDH, FCC=0.143), triosephosphate isomerase (TPI, FCC=0.136) and pyruvate kinase (PK, FCC=0.136) had a large control effect on lactate flux, while glucose-6-phosphate dehydrogenase (G6PDH, FCC=−0.246) had a negative control function. These findings will enable future detailed investigation of metabolic mechanism of fruits and research of new preservatives.

Diffusion and bulk flow in phloem loading: a theoretical analysis of the polymer trap mechanism for sugar transport in plants.

Plants create sugar in the mesophyll cells of their leaves by photosynthesis. This sugar, mostly sucrose, has to be loaded via the bundle sheath into the phloem vascular system (the sieve elements), where it is distributed to growing parts of the plant. We analyze the feasibility of a particular loading mechanism, active symplasmic loading, also called the polymer trap mechanism, where sucrose is transformed into heavier sugars, such as raffinose and stachyose, in the intermediary-type companion cells bordering the sieve elements in the minor veins of the phloem. Keeping the heavier sugars from diffusing back requires that the plasmodesmata connecting the bundle sheath with the intermediary cell act as extremely precise filters, which are able to distinguish between molecules that differ by less than 20% in size. In our modeling, we take into account the coupled water and sugar movement across the relevant interfaces, without explicitly considering the chemical reactions transforming the sucrose into the heavier sugars. Based on the available data for plasmodesmata geometry, sugar concentrations, and flux rates, we conclude that this mechanism can in principle function, but that it requires pores of molecular sizes. Comparing with the somewhat uncertain experimental values for sugar export rates, we expect the pores to be only 5%-10% larger than the hydraulic radius of the sucrose molecules. We find that the water flow through the plasmodesmata, which has not been quantified before, contributes only 10%-20% to the sucrose flux into the intermediary cells, while the main part is transported by diffusion. On the other hand, the subsequent sugar translocation into the sieve elements would very likely be carried predominantly by bulk water flow through the plasmodesmata. Thus, in contrast to apoplasmic loaders, all the necessary water for phloem translocation would be supplied in this way with no need for additional water uptake across the plasma membranes of the phloem.

Spatio-temporal dynamics of fructan metabolism in developing barley grains.

Barley (Hordeum vulgare) grain development follows a series of defined morphological and physiological stages and depends on the supply of assimilates (mainly sucrose) from the mother plant. Here, spatio-temporal patterns of sugar distributions were investigated by mass spectrometric imaging, targeted metabolite analyses, and transcript profiling of microdissected grain tissues. Distinct spatio-temporal sugar balances were observed, which may relate to differentiation and grain filling processes. Notably, various types of oligofructans showed specific distribution patterns. Levan- and graminan-type oligofructans were synthesized in the cellularized endosperm prior to the commencement of starch biosynthesis, while during the storage phase, inulin-type oligofructans accumulated to a high concentration in and around the nascent endosperm cavity. In the shrunken endosperm mutant seg8, with a decreased sucrose flux toward the endosperm, fructan accumulation was impaired. The tight partitioning of oligofructan biosynthesis hints at distinct functions of the various fructan types in the young endosperm prior to starch accumulation and in the endosperm transfer cells that accomplish the assimilate supply toward the endosperm at the storage phase.

Expression profiling of sucrose metabolizing genes in Saccharum, Sorghum and their hybrids.

Sucrose phosphate synthase (SPS; EC 2.4.1.14), sucrose synthase (SuSy; EC 2.4.1.13) and soluble acid invertase (SAI; EC 3.2.1.26) are key enzymes that regulate sucrose fluxes in sink tissues for sucrose accumulation in sugarcane and sorghum. In this study, the expression profiling of sucrose-related genes, i.e. SPS, SuSy and SAI in two sets of hybrids viz., one from a Sorghum × Saccharum cross and the other from a Saccharum × Sorghum cross, high- and low-sucrose varieties, sweet and grain sorghum lines was carried out using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) at monthly intervals. The results indicated differential expression of the three genes in high- and low-sucrose forms. Expression of SPS and SuSy genes was high in high-sucrose varieties, Saccharum × Sorghum hybrids and sweet sorghum and lower in low-sucrose varieties, Sorghum × Saccharum hybrids and grain sorghum. SAI showed a lower expression in high-sucrose varieties, Saccharum × Sorghum hybrids and sweet sorghum and higher expression in low-sucrose varieties, Sorghum × Saccharum hybrids and the grain sorghum. This study describes the positive association of SPS and SuSy and negative association of SAI on sucrose accumulation. This is the first report of differential expression profiling of SPS, SuSy and SAI in intergeneric hybrids involving sugarcane and sorghum, which opens the possibility for production of novel hybrids with improved sucrose content and with early maturity.

FLOWER ABSCISSION IN PEPPER PLANTS GROWN UNDER DIFFERENT REGIMES OF NITROGEN FERTILIZATION AND PHOTOSYNTHETICALLY ACTIVE RADIATION

The effect of nitrogen fertilization on flower abscission in pepper was studied under different growth regimes. The pepper plants were irrigated with 4, 9, and 14 meq L−1of nitrate (N4, N9, and N14). The plants were grown in winter under a low level (639 mol m−2) of cumulative photosynthetically active radiation (PAR) (LPAR), and in spring under a high level (1074 mol m−2) of cumulative PAR (HPAR). The number of flowers and flower abscission were higher under HPAR than under LPAR. Flower abscission was higher in response to treatment N4, than in response to treatments N9 and N14, while the flower number was significantly lower. Flower abscission was strongly correlated with growth-related parameters as well as with the carbon or nitrogen contents in plants. Neither the sucrose fluxes nor the amino acid fluxes through the flower pedicels were affected by nitrogen supply. The sucrose fluxes were strongly correlated with air temperature.

Laser interferometric analysis of glucose and sucrose diffusion in agarose gel.

The paper presents the investigation results of glucose and sucrose diffusion in agarose gel studied with laser interferometry method and the results of fluorescence analysis of the macroscopic gel structure. The diffusion kinetics of these substances released from aqueous solutions of a molar concentration of 0.05 M into the agarose solutions of concentrations of 0.5% and 3% in two gravitational configurations of measuring system was analysed. In the first configuration the solute diffused according, whereas in the second one - opposite to the gravitational force. The diffusion was analysed in the time period between 120 and 2400 s with a time interval of Δt = 120 s. We observed that the convective instabilities were damped well by the agarose gel, which gives the possibility of the interferometric studies of the diffusive transport for other substances in different gravitational configurations of the system. The time characteristics of glucose and sucrose fluxes in both configurations of the system and the gravitational polarisation coefficient values were obtained. The substantial differences in fluxes of glucose and sucrose diffused according and opposite to the gravitational force were observed. Additionally, we observed the differences between the diffusive fluxes of these substances in both configurations in dependence on the gel solution concentration (which is associated with gel porosity dependent on its concentration) and the kind of diffused substance.

Modeling the parameters for plasmodesmal sugar filtering in active symplasmic phloem loaders.

Plasmodesmata (PD) play a key role in loading of sugars into the phloem. In plant species that employ the so-called active symplasmic loading strategy, sucrose that diffuses into their unique intermediary cells (ICs) is converted into sugar oligomers. According to the prevalent hypothesis, the oligomers are too large to pass back through PD on the bundle sheath side, but can pass on into the sieve element to be transported in the phloem. Here, we investigate if the PD at the bundle sheath-IC interface can indeed fulfill the function of blocking transport of sugar oligomers while still enabling efficient diffusion of sucrose. Hindrance factors are derived via theoretical modeling for different PD substructure configurations: sub-nano channels, slit, and hydrogel. The results suggest that a strong discrimination could only be realized when the PD opening is almost as small as the sugar oligomers. In order to find model parameters that match the in vivo situation, we measured the effective diffusion coefficient across the interface in question in Cucurbita pepo with 3D-photoactivation microscopy. Calculations indicate that a PD substructure of several sub-nano channels with a radius around 7 Å, a 10.4 Å-wide slit or a hydrogel with 49% polymer fraction would be compatible with the effective diffusion coefficient. If these configurations can accommodate sufficient flux of sucrose into the IC, while blocking raffinose and stachyose movement was assessed using literature data. While the slit-configuration would efficiently prevent the sugar oligomers from “leaking” from the IC, none of the configurations could enable a diffusion-driven sucrose flux that matches the reported rates at a physiologically relevant concentration potential. The presented data provides a first insight on how the substructure of PD could enable selective transport, but indicates that additional factors are involved in efficient phloem loading in active symplasmic loading species.