trends in plant science Reviews

Sugar transporters in higher plants a diversity of roles and complex regulation

Lorraine E. Williams, Remi Lemoine and Norbert Sauer
Sugar-transport proteins play a crucial role in the cell-to-cell and long-distance distribution of sugars throughout the plant. In the past decade, genes encoding sugar transporters (or carriers) have been identified, functionally expressed in heterologous systems, and studied with respect to their spatial and temporal expression. Higher plants possess two distinct families of sugar carriers: the disaccharide transporters that primarily catalyse sucrose transport and the monosaccharide transporters that mediate the transport of a variable range of monosaccharides. The tissue and cellular expression pattern of the respective genes indicates their specific and sometimes unique physiological tasks. Some play a purely nutritional role and supply sugars to cells for growth and development, whereas others are involved in generating osmotic gradients required to drive mass flow or movement. Intriguingly, some carriers might be involved in signalling. Various levels of control regulate these sugar transporters during plant development and when the normal environment is perturbed. This article focuses on members of the monosaccharide transporter and disaccharide transporter families, providing details about their structure, function and regulation. The tissue and cellular distribution of these sugar transporters suggests that they have interesting physiological roles.

n higher plants, CO2 fixation occurs predominantly in mesophyll cells of mature leaves. These are net exporters of sugars and are known as carbon sources. Heterotrophic cells in roots, reproductive structures, storage and developing organs rely on a supply of sugars for their nutrition; these are known as carbon sinks (net importers). Mechanisms must exist to ensure that all sink tissues receive an adequate supply of sugars for growth and development, and sugar transporters play a pivotal role in the membrane transport of sugars and their distribution throughout the plant (Fig. 1). Long-distance transport of carbohydrates between sources and sinks occurs in specific cells of the vascular system, the phloem sieve elements (Fig. 1). During their development, sieve elements extend longitudinally, degrade most of their organelles, including vacuoles, ribosomes and nuclei, and form large, plasma membrane-lined pores in their terminal cell walls, the sieve plates. The loss of organellar structures in sieve elements is paralleled by an extreme increase in organelle density in the phloem companion cells. Sieve elements and companion cells are intimately connected by numerous branched plasmodesmata and form the socalled sieve elementcompanion cell complex (SECCC). An important function of companion cells is the supply of energy and proteins to the sieve elements. Phloem sap in sieve elements moves by bulk flow with rates of up to 1 m per h. At the source, accumulation of sugar inside the SECCC (at a concentration of several hundred mM to 1 M) results in the osmotic uptake of water, forcing sap to flow along the sieve tube. Unloading of sugar at the sink results in loss of water from the sieve tube and thus the gradient of pressure is maintained. Sucrose represents the main form of reduced carbon transported in sieve elements, although some plants can also transport raffinose, stachyose and polyols. These solutes are less subject to metabolism than glucose. After its synthesis, sucrose can pass the entire route from the mesophyll cells to the sieve

elementcompanion cell in the symplast, moving from cell to cell via plasmodesmata (symplastic loading; Fig. 1). However, frequently, sucrose is released from the mesophyll cells and actively loaded from the apoplast into the SECCC (apoplastic loading). Plasma membrane sucroseH symporters are fundamental to this process. Unloading of sucrose at the sinks might also occur either symplastically or apoplastically. The route depends on species (both pathways might operate in the same organism), organ or tissue, and developmental stage. Sink cells either import sucrose from the apoplast directly via sucrose transporters or, alternatively, sucrose can be hydrolysed to glucose and fructose by cell-wallbound invertases and taken up via monosaccharide transporters. Plants appear to have several monosaccharide and disaccharide transporters to coordinate sugar transport in diverse tissues, at different developmental stages and under varying environmental conditions. The nomenclature in the literature for plant sugar transporters has become rather confusing with SUT and SUC being used for disaccharide transporters, and STP, MST, HEX and ST being used for monosaccharide transporters. Here we will refer to disaccharide transporters as DSTs and refer to monosaccharide transporters as MSTs. Some of the physiological functions of these sugar transporters, as suggested from work at the molecular level, will be discussed. Other proteins that have been implicated in the transport of sugars [e.g. sugar-binding protein, SBP (Ref. 1)] will not be considered here.
Multi-gene families for sugar transporters

In Arabidopsis, 14 putative MST genes have been identified to date2 and at least seven DST genes can be found in Arabidopsis data libraries. Heterologous expression in yeast or Xenopus oocytes has been carried out for some of the family members, and evidence suggests that they function as proton symporters38. Generally the DSTs are highly specific for sucrose, although some have been shown to transport maltose9. Their Km values are in the
July 2000, Vol. 5, No. 7

1360 - 1385/00/$ see front matter 2000 Elsevier Science Ltd. All rights reserved. PII: S1360-1385(00)01681-2

283

trends in plant science Reviews

(Fig. 4). Only a few DSTs and MSTs have been localized precisely at the membrane level12,13. Many authors have simply assumed that these proteins are localized at the plasma membrane, because they are targeted to this membrane in transgenic yeast. Detailed studies are required to confirm this for each family member. Some of the more interesting physiological functions of a range of sugar transporters, as suggested from work at the molecular level, are discussed below.
Phloem-associated sucrose transporters

P P

Sieve elements

Water

Autotrophic source cell S ?

Companion cell

ATP H H+ S
+

Source ADP+Pi S

S H+

In apoplastic phloem loading, a DST protein at the plasma membrane of phloem cells accuS mulates sucrose in the SECCC to drive longdistance transport (Figs 1 and 4). In some Path plants these sucrose transporters have been S S detected in sieve elements (Ref. 12), whereas + H others have been observed in companion G+F cells14,15. This might be explained by species + + H G S H differences, but recent work in Plantago has shown that there are sucrose transporters speInvertase cific to either companion cells or sieve elS S ements (R. Stadler et al., unpublished). All of these transporters result from gene transcripP tion in the companion cell, and it will be Sink Heterotrophic important to determine the signals and mechsink cell anisms that regulate their final destination. In Solanaceae, SUT1 (DST) mRNA is localized in the sieve elements, suggesting that tranWater scription occurs in the companion cell and Trends in Plant Science might be followed by the transportation of mRNA through the plasmodesmata and transFig. 1. The contribution of sugar transporters to long-distance sugar transport between sources lation within the sieve elements12. and sinks. Long-distance transport of sucrose (S) between sources and sinks occurs in sieve In Solanaceous species, antisense studies elements of the phloem, which constitute part of the vascular system (path). Sucrose is transported in the phloem sap by bulk flow. In the source leaf, sucrose can pass the entire route from have shown that SUT1 (DST) is indeed the mesophyll cells (source cell) to the sieve elementcompanion cell in the symplast, moving essential for phloem loading and long-disfrom cell to cell via plasmodesmata (P). However, in many species, sucrose leaves the symplast tance transport16,17. Potato plants with drastiat some point, possibly via sucrose efflux carriers (red circle), and is actively accumulated from cally lowered expression of SUT1 (DST) the apoplast into sieve elements and/or companion cells by plasma membrane protonsucrose had a reduced tuber yield (size and number), symporters (purple and dark-blue circles, respectively). Energization is via the proton-motive and mature leaves accumulated sugars force generated by the plasma membrane H-ATPase (black square). Passive influx of water because of impaired export16,17. This was into the sieve tubes presumably occurs via water channels. Expression of sucrose carriers confirmed in tobacco, where strong anti(light-green circle) along the path has been related to a function for retrieving sucrose leaked sense plants had a dwarf phenotype, and from the phloem. Unloading of sucrose into sink cells might occur symplastically via plassugar export from leaves was also drastimodesmata or sucrose might be delivered to the apoplast via a sucrose efflux carrier (pink circle). Here, it is either taken up directly by plasma membrane sucrose transporters (yellow cally impaired18. In wild-type plants, circle), or first hydrolysed to glucose (G) and fructose (F) by the cell-wall invertase (dark-green lengthy dark exposure leads to starch degraelipse) then taken up via plasma membrane monosaccharide transporters (light-blue circle). dation in leaves and subsequent exportation of sucrose to sink organs. However, in antisense plants, starch remains in the leaves 0.31.0 mM range9. The MSTs have broader substrate specificity, and this is related to impaired sucrose export. Phloem-localized transporting a range of hexoses and pentoses with Km values for the SUT1 (DST) homologues might play an additional role in both the preferred substrate being typically 10100 M (Ref. 2). MSTs and retrieval of sucrose leaking from sieve elements along the transloDSTs share little homology at the amino acid level, although there cation pathway and possibly in catalysing the export of sucrose are structural similarities (Fig. 2). They are thought to be members of from the phloem in sinks19 (Fig. 1). Moreover, the same transporters the major facilitator superfamily10,11. Whereas DST genes appear to are not always confined to the phloem but are also present at lower be specific for plants, numerous genes that are homologous to the levels in other leaf cells where they might function in retrieval17,20. MSTs are found in bacteria, fungi and mammals (Fig. 3).
Sugar transporters in roots
Diverse locations for sugar transporters

Plant cells appear to possess sugar transporters tailored to meet their specific requirements at particular stages of development
284
July 2000, Vol. 5, No. 7

Roots represent heterotrophic carbon sinks and can be divided into zones of cell division, elongation and maturation. AtSTP4 (MST), a monosaccharide transporter from Arabidopsis, is found

trends in plant science Reviews

only at the root tip, probably in the root meristem21 (Fig. 4). This indicates that part of the post-phloem transport in Arabidopsis roots occurs apoplastically and that the role of AtSTP4 (MST) is to import hexoses generated from sucrose hydrolysis by cell-wall invertase following phloem unloading. In Medicago truncatula, the gene for the monosaccharide transporter, Mtst1 (MST), is only expressed in cells immediately behind the root apical meristem, in cells within the elongation zone and possibly in the zone of cell division22. Elongating cells have a high requirement for hexoses as precursors for macromolecular synthesis and to allow them to maintain osmotic pressure during elongation22. Mtst1 (MST) is also expressed in primary phloem fibres where it might provide hexoses for cell wall biosynthesis and deposition22. Sucrose transporters have also been detected in particular cells of the root: DcSUT2 (DST) is expressed in storage parenchyma tissues of carrot tap-roots where it seems to import sucrose for storage23. This and other DST homologues have also been detected in phloem cells of the root, where they might retrieve sucrose into the phloem or possibly catalyse the efflux of sucrose from the phloem19.
A role for sugar transporters in developing and germinating seeds

(a)

Disaccharide-H+-symporter (PmSUC2)

400 50 120 220 300

470

170 90 270

350

440

510

Cytoplasmic side

(b)
50

Monosaccharide-H+-symporter (NtMST1)

380 80 140 310 200 450

20

The developing embryo is symplastically iso410 lated from maternal tissues and is reliant on 480 230 the import of sugars from the surrounding apoplastic space. Sugars are required for embryo development and for the deposition 1 of storage compounds necessary for germi270 nation. DSTs and MSTs have been identified Cytoplasmic side in Vicia faba, Pisum sativum and Hordeum vulgare (Refs 2426). In Vicia, VfSTP1 523 (MST) is expressed in the embryo cotyledon epidermal cells. Expression is confined to epiTrends in Plant Science dermal regions covering the mitotically active Fig. 2. Topological models of the disaccharideHsymporter, (a) PmSUC2 [disaccharide parenchyma, indicating that this transporter 24 transporter (DST)] and (b) the monosaccharideH symporter, NtMST1 [monosaccharide serves to supply substrate for metabolism . transporter (MST)]. MSTs and DSTs have predicted molecular masses of ~55 kDa and During later stages of embryo development, hydrophobicity analysis indicates that they are highly hydrophobic integral membrane proVfSTP1 (MST) is replaced by the sucrose teins. They share little homology at the amino acid level, although there are structural simi24 transporter VfSUT1 (DST) . VfSUT1 (DST) larities and they are thought to be members of the major facilitator superfamily. In this family, expression is highest in epidermal cells with it is proposed that the twelve transmembrane -helical structure has evolved from an ancestransfer-cell morphology and with storage tral six-transmembrane-segment transporter gene following gene duplication and fusion10. activity in the underlying parenchyma, indiFamily members generally possess a large central loop between transmembrane domains six and seven (the length of this loop varies in members of the DST and MST families). cating that the encoded transporter is responsible for providing substrate for the synthesis of storage compounds24. Certain seeds accumulate storage reserves in the endosperm dur- is transferred symplastically to the phloem region. The high expresing development. Upon germination, these are hydrolysed and the sion of RcSUT1 (DST) in the phloem indicates that sugars are products, including sugars, are released into the apoplastic space released into the apoplast before active loading by this transporter. (Fig. 5). In germinating seeds of Ricinus communis, a sucrose carrier, RcSUT1 (DST), is expressed at high levels in both the coty- A variety of roles for sugar transporters in floral organs ledon epidermal cells that are situated adjacent to the endosperm Developing pollen grains are strong sinks that require carbohyand also in the phloem13. This suggests that the epidermal cells ini- drate import from the apoplast during maturation, germination tially accumulate sucrose from the apoplast and from here sucrose and growth. The Arabidopsis monosaccharide transporter gene,
July 2000, Vol. 5, No. 7

285

trends in plant science Reviews

(a)
EcXyIE SspGTR
100

(b)
EcAraE Prokaryotes EcGaIP AtERD6 HsGLUT1 Mammal ScHXT2 100 46 ScGAL2 98 ScHXT1 100 99 Fungi ScHXT3 AmMST1 96 ScSNF3 100 ScRGT2 CkHUP2 97 CkHUP1 Lower plants 100 CkHUP3 NtMST1 VvMST1 42 48 RcSTC 30 AtSTP1 100 18 AtSTP12 VfMST1 99 95 MtMST1 AtSTP4 76 PhPMT1 100 64 AtSTP11 78 99 AtSTP10 100 AtSTP9 RcHEX6 100 77 AtSTP3 SspSGT2 100 RcHEX1 90 AtSTP7 89 97 PaMST1 100 AtSTP14 81 AtSTP13 100 LeMST1 AtSTP2 75 AtSTP6 100 AtSTP8 EcMeIB AtSUC3
100 100 54 99 97

100

Arabidopsis
Monocots

20

HvSUT1 OsSUT1

71

HvSUT2 DcSUT1 AtSUC4

100

Arabidopsis

PmSUC1
100

40

21 84 95 56 29 100

BvSUT1 SoSUT1 AmSUT1

89

AbSUT1 StSUT1 NtSUT1

PmSUC2

Plants with symplastic phloem loading

VfSUT1

Higher plants

35 71 100

AtSUC2 AtSUC13
100 84 66

AtSUC8 AtSUC1 AtSUC5

Arabidopsis

100

DcSUT2 AgSUT1

Trends in Plant Science

Fig. 3. Phylogenetic analysis for (a) monosaccharide transporters (MSTs) and (b) disaccharide transporters (DST). (a) Sugar-transport proteins homologous to lower and higher plant monosaccharide transporters are found in prokaryotes, fungi and mammals, indicating that the corresponding genes form an ancient gene family. Monosaccharide transporters from one species, such as the Arabidopsis AtSTPs, do not form a single subcluster within the higher plant subfamily, suggesting that several independent genes already existed at the beginning of or early in higher plant evolution. The less closely related Arabidopsis AtERD6 protein (Accession no. D89051), described as a putative sugar transporter, clusters with the human glucose facilitator. (b) Proteins closely related to higher plant sucrose transporters are not found in bacteria, fungi or mammals, indicating that plant sucrose-transporter genes evolved comparatively late. Sucrose transporters from Arabidopsis cluster together with sucrose transporters from monocots and from plants with symplastic phloem loading. A functionally uncharacterized transporter from Schizosaccharomyces pombe (Accession no. Z99165) shows some similarity, but only in the N-terminal half of the protein. EcMelB (the outgroup) is extremely distantly related to plant sucrose transporters45. Bootstrap values given at internal nodes indicate the percentage of the occurence of these nodes in 100 replicates (maximum parsimony) of the data set. The sequence of the E. coli xylose permease (EcXylE; black) and of the E. coli melibiose transporter (EcMelB; black) were used as outgroups in (a) and (b), respectively. Accession numbers for nonplant monosaccharide transporter sequences are AmMST1 (Amanita muscaria monosaccharide transporter), Z83828; EcAraE (E. coli arabinose transporter), J03732; EcGalP (E. coli galactose permease), AE000377; EcMelB (E. coli melibiose transporter), K01991; EcXylE (E. coli xylose transporter), P09098; HsGlut1 (Homo sapiens glucose transporter), 87529; ScHXT1 (Saccharomyces cerevisiae hexose transporter), M82963; ScHXT2 (S. cerevisiae hexose transporter), M33270; ScHXT3 (S. cerevisiae hexose transporter), L07080; ScRGT2 (S. cerevisiae hexose transporter-like glucose sensor), Z74186; ScSNF3 (S. cerevisiae hexose transporter-like glucose sensor), J03246; SspGTR (Synechocystis sp. glucose transporter), X16472. Maximum parsimony (MP) bootstrap analyses of aligned sequences were conducted with the PHYLIP package 3.572c. Addition of taxa was jumbled and repeated three times for each individual bootstrap replication in the MP analysis. Output treefiles were combined with the CONSENSE program and converted into a graphical representation by the TREEVIEW program. Accession numbers of plant sucrose transporters used in this tree are AbSUT1 (Asarina barclaiana, AF191024); AgSUT1 (Apium graveolens, AF063400); AmSUT1 (Alonsoa meridionalis, AF191025); AtSUC1 (Arabidopsis thaliana, X75365); AtSUC2 (A. thaliana, X75382); AtSUC3 (A. thaliana, AC004138); AtSUC4 (A. thaliana, AC000132); AtSUC5 (A. thaliana, AJ252133); AtSUC8 (A. thaliana, AAC69375); AtSUC13 (A. thaliana, AB016875); BvSUT1 (Beta vulgaris, X83850); DcSUT1 (Daucus carota, Y16766); DcSUT2 (D. carota, AB036758); HvSUT1 (Hordeum vulgare, Accession no. not yet available); HvSUT2 (H. vulgare, Accession no. not yet available); NtSUT1 (Nicotiana tabacum, X82276); OsSUT1 (Oryza sativa, D87819); PmSUC1 (Plantago major, X75764); PmSUC2 (P. major, X75764); SoSUT1 (Spinacea oleracea, X67125); StSUT1 (Solanum tuberosum, X69165); VfSUT1 (Vicia faba, Z93774). Accession nos of plant monosaccharide transporters used in this tree are AtSTP1 (A. thaliana, X55350); AtSTP2 (A. thaliana, AJ001362); AtSTP3 (A. thaliana, AJ002399); AtSTP4 (A. thaliana, X66857); AtSTP6 (A. thaliana, AC013454); AtSTP7 (A. thaliana, AF001308); AtSTP8 (A. thaliana, AF077407); AtSTP9 (A. thaliana, AC007980); AtSTP10 (A. thaliana, AB025631); AtSTP11 (A. thaliana, AB007648); AtSTP12 (A. thaliana, AL022603); AtSTP13 (A. thaliana, AF077407); AtSTP14 (A. thaliana, AC004260); CkHUP1 (Chlorella kessleri, Y07520); CkHUP2 (C. kessleri, X66855); CkHUP3 (C. kessleri, X75440); LeMST1 (Lycopersicum esculentum, AJ010942); MtMST1 (Medicago truncatula, U38651); PaMST1 (Picea abies, Z83829); PhPMT1 (Petunia hybrid, AF061106); RcHEX1 (Ricinus communis, L08197); RcHEX6 (R. communis, L08188); RcSTC (R. communis, L08196); SspSGT2 (Saccharum hybrid, L21753); VfMST1 (V. faba, Z93775); VvMST1 (Vitis vinifera, AJ001061).

286

July 2000, Vol. 5, No. 7

trends in plant science Reviews

AtSTP2 (MST), is expressed after meiosis of the pollen mother cell at the time when the uninucleate microspores are develop(e) ing27. This correlates with the time when callose surrounding the microspores is Papilla (d) degraded. After this, AtSTP2 (MST) Pollen tube 27 mRNA and protein are rapidly degraded . Thus, AtSTP2 (MST) could serve in the Transmitting uptake of glucose (resulting from callose tissue degradation) and also other monosaccharides such as xylose, mannose or galactose (from degradation of other cell-wall components) into the symplastically isolated, Pollen Pollen developing male gametophyte27. sac Carbohydrates are also required to supConnective ply energy for pollen germination and (a) (b) tissue pollen-tube growth. PhPMT1 (MST), a putative hexose transporter gene in Petunia, is expressed specifically in mature Xylem anthers and pollen, and not in any other flovessel ral or vegetative tissues28. Thus, in this Companion species, sucrose might be hydrolysed to cells glucose and fructose by a cell-wall invertase before uptake by PhPMT1 (MST) in the growing pollen tube28. A similar local(c) ization has been shown for the Arabidopsis AtSTP4 (MST) monosaccharide transporter21 (Fig. 4); however, its function is unclear because in this species (and in many others) sucrose is the preferred carbon source during pollen germination and growth in vitro. Recent studies describing the expression of the sucrose transporter genes, AtSUC1 (DST) in germinating Arabidopsis pollen (Fig. 4) and NtSUT3 (DST) in tobacco pollen, are consistent with this29,30. Interestingly, AtSUC1 (DST) mRNA was detected in developing pollen whereas the protein was only found after pollen germination, indicating that expresFig. 4. Tissue and cellular distribution of various sugar transporters. The tissue or cell-specific sion might be translationally regulated and expression of several sugar transporters indicates their specific physiological tasks. For example, induced during germination29. sugar transporters have been implicated in processes as diverse as phloem loading and unloading, AtSUC1 (DST) is also expressed in a pollen development and germination, and pathogen interaction. (a) Expression of the sucrose ring of parenchymatic cells surrounding transporter gene AtSUC2 (disaccharide transporter, DST) in the veins of Arabidopsis source leaves the transmitting tissue of the style and in indicates a role in phloem loading. The expression is demonstrated by the activity of the reporter 29 the epidermal cell layers of the funiculi . gene for -glucuronidase (GUS) driven by the AtSUC2 promoter19. In (b), the AtSUC2 (DST) This expression pattern is unlikely to protein was immunolocalized with fluorescent secondary antibodies in companion cells of leaf reflect a higher requirement for carbohyminor veins15. Xylem vessels are identified by the yellow autofluorescence of lignin. (c) Expression of the monosaccharide transporter gene, AtSTP4, from Arabidopsis in root tips is shown drates by these cells; instead it might indiby GUS expression under the control of the AtSTP4 promoter21. The expression of this carrier is cate that AtSUC1 (DST) modulates the also induced by wounding and pathogen attack. (d) In situ hybridization of AtSUC1 (DST) mRNA availability of water in this region, which on an anther cross-section29. At this stage, the AtSUC1 (DST) protein is detected in the connective could establish a cue for pollen-tube guidtissue but not in the pollen, suggesting translational regulation. The sucrose carrier AtSUC1 (DST) ance towards the ovule29. mRNA from Arabidopis has been identified in the connective tissue of mature anthers and also In anthers, AtSUC1 (DST) mRNA and in pollen. In the connective tissue, AtSUC1 (DST) could be involved in the control of anther protein were detected in parenchymatic dehiscence. (e) The monosaccharide transporter AtSTP4 (MST) is detected in pollen tubes cells surrounding the central vascular bungrowing inside the transmitting tissue of the style (longitudinal section through a style). Specific dle of anther connective tissue just before antibodies were used and the fluorescence of the secondary antibodies appears in green. 29 anther opening by dehiscence (Fig. 4). Interestingly, the sucrose transporter AtSUC1 (DST) is also present in germinating pollen29. Accumulation of sucrose by AtSUC1 (DST) could decrease the water potential and hence result in water uptake from the adjacent cells of the Sugar transporters in plantmicroorganism interactions anther walls29. This would result in dehydration of the endothecium Some sugar transporters seem to play a role in certain and an increased tension of its wall thickenings, finally causing plantmicroorganism interactions. The sink-specific, woundanther opening29. induced monosaccharide transporter, AtSTP4 (MST) is induced
July 2000, Vol. 5, No. 7

287

trends in plant science Reviews

cope with the increased demand related to defence activities, or serve to recover sugars from the apoplast (particularly if hostcell permeability is increased) and thus reduce the loss of carbohydrate to the pathogen. Sugar transporters are also important in carbon transfer to mycorrhizal fungi22. Colonization of Medicago truncatula roots with the mycorrhizal fungus Glomus versiforme, induces expression of the monosaccharide transporter gene, MtST1 (MST), in cortical cells of the root that correlates with regions that are heavily colonized by the fungus22. The effect is localized to cells within the immediate vicinity of the fungus, either in cells containing an arbuscule, or cells that are frequently in contact with intercellular hyphae. The nature of the signal for the induction of plant sugar-carriers during microorganism interactions is not yet clear.
Role in sugar sensing

(a)

(b)
Lipids Sucrose
Co

tyl co po y H

t yle

ns do Proteins

Endosperm

Amino acids

(c) (i)
LE

(ii) LE

(iii) LE

Sugars can act as regulatory signals that affect gene expression and hence plant development. In relation to resource allocation, the ability to sense altered sugar concentrations could have important advantages by allowing the plant to tailor its metabolism in source tissues to meet the demands in sinks. There is great interest in Fig. 5. Localization of the sucrose carrier in a germinating Ricinus seedling. Seeds accudissecting the signalling processes that are mulate storage reserves in the endosperm during development. Upon germination, these are involved in sugar sensing and response hydrolysed and the products, including sugars, are released into the apoplastic space. Sevenpathways. There are three possible sugarday-old dark-grown germinating Ricinus seedlings are shown in (a) with (left) and without sensing systems in plants32: (right) an endosperm. (b) Breakdown of lipids and protein in the endosperm and the release of A hexokinase-sensing system. sucrose and amino acids into the apoplast for uptake by the cotyledon. (c) Expression pattern A sucrose-transport-associated sensor. of the sucrose transporter, RcSUT1 (DST)13,46, in the germinating Ricinus seedling (three-day A glucose-transport-associated sensor. old). RcSUT1 transcripts were localized by in situ hybridization using an antisense RcSUT1 cRNA probe. Dark staining, indicating RcSUT1 expression, is seen in both the cotyledon Based on sugar signal transduction processes lower epidermal cells (LE) that are situated adjacent to the endosperm (parts iiii) and also in in Saccharomyces, it has been proposed that the phloem (red arrows in iii). No equivalent staining was observed in sections probed with members of the sugar transporter family in the sense probe (results not shown). The expression pattern indicates that the epidermal cells plants might play a role in sugar sensing33. In initially accumulate sucrose from the apoplast and from here sucrose is transferred symplastiyeast, it has been shown that membranecally to the phloem region. The high level of expression of RcSUT1 in the phloem indicates bound receptors closely related to sugar that sugars are released into the apoplast before active loading by this transporter. Scale bars transporters are used to sense external sugar 100 m in (i) and (ii); 50 m in (iii). (c) reproduced, with permission, from Ref. 13. concentrations and activate a signal transduction pathway leading to the regulation of approximately fourfold in Arabidopsis seedlings infected with transporter gene expression and insertion or degradation of proteins Alternaria brassicicola or Fusarium oxysporum, and also in sus- at the plasma membrane. This allows yeast to respond to a rapidly pension-cultured cells of Arabidopsis treated with bacterial or changing external environment. The yeast glucose sensors, ScSNF3 fungal elicitors21. It has been concluded that AtSTP4 (MST) plays (senses low glucose levels) and ScRGT2 (senses high glucose leva role in the transportation of monosaccharides into the sink els) are homologous to yeast and plant glucose transporters (Fig. 3) tissues and that it responds to an increased demand for carbohy- but do not mediate significant glucose transport34. The large Cdrate by cells under stress conditions21. Induction of AtSTP4 terminal extensions of these proteins are required for the trans(MST) expression is also observed during plant and fungal mission of the glucose signal. To date, there is no direct evidence for biotrophic interaction, for example, Arabidopsis infected with a role of plant monosaccharide and disaccharide transporters as the powdery mildew Erysiphe cichoracearum31. Expression of sensors. Nevertheless, the finding that several members of the plant AtSTP4 (MST) is induced dramatically in source leaves infected MST and DST families have unusual cytoplasmic extensions, which with mildew and, although the fungus is confined to epidermal are predicted to be cytosolic, has initiated studies in relation to the cells, AtSTP4 (MST) expression is induced in many cell types possible role of these proteins in sugar sensing33. within the leaf following infection (M. Gilbert et al., unpublished). The signalling pathway by which this occurs is unknown, and the Regulation of sugar transporters role of this transporter in this interaction is unclear. It might help As sugar transporters play such a key role in sourcesink interacto increase carbohydrate import into infected tissues, in order to tions, it is likely that they are tightly controlled. There are several
288
July 2000, Vol. 5, No. 7

trends in plant science Reviews

levels at which regulation might occur35,36. As for all H-symporters, regulation of activity by the proton-motive force is clearly important. Thus, modulations in the activity of the H-ATPase would immediately affect their sugar transport kinetics. There are numerous examples for transcriptional regulation of DST or MST gene expression, such as changes in DST expression pattern in leaves undergoing sink-to-source transition19,37 during seed development (MST and DST)24, or during pollen maturation and germination (MST and DST)27,28. There is also evidence that biotic and abiotic factors (e.g. mechanical treatments, light, water and salt stress, phytohormones, and pathogen attack) have effects on the expression of certain sugar transporters21,36,38. Post-transcriptional control mechanisms such as mRNA stability (turnover), mRNA translation and post-translational control have also been described for sugar transporters. Examples include the diurnal regulation of DSTs (Refs 12,39), phosphorylationdephosphorylation40, and possible modification of activity by the lipid environment36. There is evidence that changing sugar levels might have an effect on sugar transporter expression and activity. Evidence has been provided that a putative sucrose-sensing pathway could modulate the expression levels and activity of a protonsucrose transporter as a function of changing sucrose concentrations in the leaf (hexoses do not elicit the response)41. This would have a direct impact on assimilate partitioning at the level of phloem translocation. Changes in transcriptional activity or mRNA turnover mediate this repression by sucrose but additional regulation by alterations in protein turnover or direct modification cannot be ruled out. The response is reversible, highlighting the dynamic nature of this signalling pathway. Evidence for sugar regulation of transporters is variable. AtSUC2 (DST) expression (analysed using a SUC2 promoter GUS construct) appears unaffected by sugars19. However, downregulation might not be seen in these experiments because of the stability of the GUS protein. In addition, it is still possible that activity might be affected by other mechanisms, even if the promoter is insensitive to sugars. The expression of the Vicia faba sucrose carrier gene, VfSUT1 (DST), but not the glucose transporter, VfSTP1 (MST), decreased in cotyledons following exposure to high concentrations of sucrose or glucose (150 mM), whereas lower levels (10 mM) had no effect on transcript levels24. Sucrose-feeding experiments with detached maize leaves led to an increase in transcript leaves of ZmSUT1 (Ref. 42) (DST). By contrast, no evidence was found for sugar regulation of three monosaccharide-transporter genes (CST13) (MSTs) in suspensioncultured cells of Chenopodium rubrum43. Thus, for sugar transporters, it is now emerging that various levels of control are implemented that allow plant cells to regulate the fluxes of sugars across the plasma membrane during normal growth and development, and also when their natural environment is perturbed.
The future

affecting sourcesink relations (e.g. following application of phytohormones and elicitors, pathogen attack and wounding)44 but further work is required to investigate the coordination of events. In sink cells, it is important to understand why, in some cases, sucrose is cleaved by a cell-wall invertase and therefore hexose carriers are involved, whereas in other cases sucrose is taken up and cleaved inside the cells. In germinating pollen, both monosaccharide and disaccharide transporters are present2830, although sucrose is the preferred carbon source for in vitro germination and hexoses are inhibiting. Clearly, work is needed for a full comprehension of the respective role of each carrier type in such cells. We need to clarify which mechanisms exist for the efflux of sugars from cells, for example, for efflux from the mesophyll or from apoplastically unloading phloem cells. Under nonenergized conditions, influx carriers might be operating in efflux, allowing sugars to be transported down their concentration gradient; alternatively, different transporters might be involved. In addition, the identification of tonoplast sugar transporters is important for understanding storage and remobilization from the vacuole. We must also consider that entirely different mechanisms might exist for sugar transport, such as those involving ABC transporters. More evidence is required on the physiological function of sugar transporters to support the information from expression studies. In particular, no antisense studies have been reported for any of the monosaccharide transporters. We eagerly await studies on knockout mutants to dissect the transport processes in greater detail. The phenotype of such plants will provide clues concerning the physiological role and importance of particular transporters. In addition, plants with transporter genes under the control of cellspecific promoters will enable us to learn more about sourcesink interactions and pathways of sugar movement. Sensing functions are essential in allowing plants to adapt to changing environments. One of the major challenges will be to identify sugar-sensing mechanisms and the signal transduction pathways. It will be important to ascertain whether members of the sugar-transporter families are involved in this function.
Acknowlegements

Our work has been supported by a grant from the European Commission DG XII Biotechnology programme (ContractBIO4-CT960583). L.E.W. is grateful to the Royal Society for support. R.L.s work is supported by the CNRS, the French Ministry for Education and Research and by the Rgion Poitou Charentes. N.Ss work was supported by the Deutsche Forschungsgemeinschaft. The help of Volker Huss with Figure 3 and the help of Cyril Maingourd with Figure 4 is gratefully acknowledged.
References 1 Overvoorde, P.J. et al. (1996) A soybean sucrose binding protein independently mediates nonsaturable sucrose uptake in yeast. Plant Cell 8, 271280 2 Bttner, M. and Sauer, N. (2000) Monosaccharide transporters in plants: structure, function and physiology. Biochim. Biophys. Acta. 1465, 263272 3 Sauer, N. et al. (1990) Primary structure, genomic organisation and heterologous expression of a glucose transporter from Arabidopsis thaliana. EMBO J. 9, 30453050 4 Sauer, N. and Stolz, J. (1994) SUC1 and SUC2: two sucrose transporters from Arabidopsis thaliana; expression and characterization in bakers yeast and identification of the histidine-tagged protein. Plant J. 6, 6777 5 Boorer, K.J. et al. (1994) Steady-state and presteady-state kinetics of the Hhexose cotransporter (STP1) from Arabidopsis thaliana expressed in Xenopus oocytes. J. Biol. Chem. 269, 2041720424 6 Boorer, K.J. et al. (1996) Transport mechanism of the cloned H/sucrose cotransporter StSUT1. J. Biol. Chem. 271, 2513925144

To comprehend sourcesink regulation fully and to be in a position to manipulate the complex interactions for agricultural benefit, it is vital that the underlying membrane transport processes are thoroughly characterized. This includes determining precisely the range of sugar transporters that are involved, their structurefunction relationships, and defining their cellular and temporal expression pattern. A particular goal would be the ability to manipulate competition between sinks. For this we need to find out how sugar carriers are regulated in metabolic and storage sinks and to relate this to the information available on the activity of the sucrose metabolizing enzymes, invertase and sucrose synthase. Extracellular invertase is up-regulated in several situations

July 2000, Vol. 5, No. 7

289

trends in plant science Reviews

7 Stolz, J. et al. (1994) Functional reconstitution of the solubilized Arabidopsis thaliana STP1 monosaccharideH symporter in lipid vesicles and purification of the histidine tagged protein from transgenic Saccharomyces cerevisiae. Plant J. 6, 225233 8 Zhou, J.J. et al. (1997) A kinetic model with ordered cytoplasmic dissociation for SUC1, an Arabidopsis H/sucrose transporter expressed in Xenopus oocytes. J. Membr. Biol. 159, 113125 9 Lemoine, R. (2000) Sucrose transporters in plants: update on function and structure. Biochim. Biophys. Acta 1465, 246262 10 Marger, M.D. and Saier, M.H., Jr (1993) A major superfamily of transmembrane facilitators that catalyse uniport, symport and antiport. Trends Biochem. Sci. 18, 1320 11 Saier, M.H. et al. (1999) The major facilitator superfamily. J. Mol. Microbiol. Biotechnol. 1, 257279 12 Khn, C. et al. (1997) Macromolecular trafficking indicated by localization and turnover of sucrose transporters in enucleate sieve elements. Science 275, 12981300 13 Bick, J.A. et al. (1998) Expression analysis of a sucrose carrier in the germinating seedling of Ricinus communis. Plant Mol. Biol. 38, 425435 14 Stadler, R. et al. (1995) Phloem loading by the PmSUC2 sucrose carrier from Plantago major occurs into companion cells. Plant Cell 7, 15451554 15 Stadler, R. and Sauer, N. (1996) The Arabidopsis thaliana AtSUC2 gene is specifically expressed in companion cells. Bot. Acta 109, 299306 16 Riesmeier, J.W. et al. (1994) Evidence for an essential role of the sucrose transporter in phloem loading and assimilate partitioning. EMBO J. 13, 17 17 Khn, C. et al. (1996) Companion cell-specific inhibition of the potato sucrose transporter SUT1. Plant Cell Environ. 19, 11151123 18 Brkle, L. et al. (1998) The H-sucrose cotransporter NtSUT1 is essential for sucrose export from tobacco leaves. Plant Physiol. 118, 5968 19 Truernit, E. and Sauer, N. (1995) The promoter of the Arabidopsis thaliana SUC2 sucrose-H symporter gene directs expression of b-glucuronidase to the phloem: evidence for phloem loading and unloading by SUC2. Planta 196, 564570 20 Lemoine, R. et al. (1996) Antisense inhibition of the sucrose transporter in potato: effects on amount and activity. Plant Cell Environ. 19, 11241131 21 Truernit, E. et al. (1996) The sink-specific and stress-regulated Arabidopsis STP4 gene: enhanced expression of a gene encoding a monosaccharide transporter by wounding, elicitors, and pathogen challenge. Plant Cell 8, 21692182 22 Harrison, M.J. (1996) A sugar transporter from Medicago truncatula: altered expression pattern in roots during vesicular-arbuscular (VA) mycorrhizal associations. Plant J. 9, 491503 23 Shakya, R. and Sturm, A. (1998) Characterization of source- and sink-specific sucrose/H symporters from carrot. Plant Physiol. 118, 14731480 24 Weber, H. et al. (1997) A role for sucrose transporters during seed development: molecular characterization of a hexose and a sucrose carrier in fava bean seeds. Plant Cell 9, 895908 25 Tegeder, M. et al. (1999) Sucrose transport into developing seeds of Pisum sativum L. Plant J. 18, 151161 26 Weschke, W. et al. (2000) Sucrose transport into barley seeds: molecular characterization of two transporters and implications for seed development and starch accumulation. Plant J. 21, 455467 27 Truernit, E. et al. (1999) A male gametophyte-specific monosaccharide transporter in Arabidopsis. Plant J. 17, 191201 28 Ylstra, B. et al. (1998) Hexose transport in growing petunia pollen tubes and characterization of a pollen-specific, putative monosaccharide transporter. Plant Physiol. 118, 297304 29 Stadler, R. et al. (1999) The AtSUC1 sucrose carrier may represent the osmotic driving force for anther dehiscence and pollen tube growth in Arabidopsis. Plant J. 19, 269278 30 Lemoine, R. et al. (1999) Identification of a pollen-specific sucrose transporter-like protein NtSUT3 from tobacco. FEBS Lett. 454, 325330 31 Hall, J.L. and Williams, L.E. Assimilate transport and partitioning in fungal biotrophic interactions. Aust. J. Plant Physiol. (in press) 32 Smeekens, S. and Rook, F. (1997) Sugar sensing and sugar-mediated signal transduction in plants. Plant Physiol. 115, 713 33 Lalonde, S. et al. (1999) The dual function of sugar carriers: transport and sugar sensing. Plant Cell 11, 707726 34 zcan, S. et al. (1998) Glucose sensing and signaling by two glucose receptors in the yeast Saccharomyces cerevisiae. EMBO J. 17, 25662573 35 Bush, D.R. (1999) Sugar transporters in plant biology. Curr. Opin. Plant Biol. 2, 187191 36 Delrot, S. et al. (2000) Regulation of sugar, amino acid and peptide plant membrane transporters. Biochim. Biophys. Acta 1465, 281306 37 Riesmeier, J.W. et al. (1993) Expression of the sucrose transporter from potato correlates with the sink-to-source transition in leaves. Plant Cell 5, 15911598 38 Noiraud, N. et al. (2000) The sucrose transporter of celery. Identification and expression during salt stress. Plant Physiol. 122, 14471455 39 Hirose, T. et al. (1997) cDNA cloning and tissue-specific expression of a gene for sucrose transporter from rice (Oryza sativa, L.). Plant Cell Physiol. 38, 13891396 40 Roblin, G. et al. (1998) Regulation of a plant plasma membrane sucrose transporter by phosphorylation. FEBS Lett. 424, 165168 41 Chiou, T.J. and Bush, D.R. (1998) Sucrose is a signal molecule in assimilate partitioning. Proc. Natl. Acad. Sci. U. S. A. 95, 47844788 42 Aoki, N. et al. (1999) Molecular cloning and expression analysis of a gene for a sucrose transporter in maize (Zea mays L.). Plant Cell Physiol. 40, 10721078 43 Roitsch, T. and Tanner, W. (1994) Expression of a sugar-transporter gene family in a photoautotrophic suspension culture of Chenopodium rubrum L. Planta 193, 365371 44 Roitsch, T. (1999) Sourcesink regulation by sugar and stress. Curr. Opin. Plant Biol. 2, 198206 45 Naderi, S. and Saier, M.H. (1996) Plant sucrose:Hsymporters are homologous to the melibiose permease of Escherichia coli. Mol. Microbiol 22, 389391 46 Weig, A. and Komor, E. (1996) An active sucrose carrier (Scr1) that is predominantly expressed in the seedling of Ricinus communis L. J. Plant Physiol. 147, 685690

Lorraine Williams* is at the School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton, UK SO16 7PX; Remi Lemoine is at the Laboratoire de Biochimie et Physiologie Vgtales, ESA CNRS 6161, Btiment Botanique, 40 Avenue du Recteur Pineau, F-86022 Poitiers Cedex, France (tel 33 5 49454183; fax 33 5 49454186; e-mail remi.lemoine@campus.univ-poitiers.fr); Norbert Sauer is at the Dept of Botany II, Molecular Plant Physiology, University of Erlangen, Staudtstrasse, D-91058 Erlangen, Germany (tel 49 9131 85 28212; fax 49 9131 85 28751; e-mail nsauer@biologie.uni-erlangen.de). *Author for correspondence (tel 44 2380 594278; fax 44 2380 594319; e-mail lew@soton.ac.uk)

Students
Did you know that you are entitled to a 50% discount on a subscription to Trends in Plant Science? See the subscription order card in this issue for details.

290

July 2000, Vol. 5, No. 7