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Afoufa-Bastien et al. BMC Plant Biology 2010, 10:245
http://www.biomedcentral.com/1471-2229/10/245

RESEARCH ARTICLE

Open Access

The Vitis vinifera sugar transporter gene family:
phylogenetic overview and macroarray
expression profiling
Damien Afoufa-Bastien1†, Anna Medici1†, Julien Jeauffre1,2, Pierre Coutos-Thévenot1, Rémi Lemoine1,
Rossitza Atanassova1, Maryse Laloi1*

Abstract
Background: In higher plants, sugars are not only nutrients but also important signal molecules. They are
distributed through the plant via sugar transporters, which are involved not only in sugar long-distance transport
via the loading and the unloading of the conducting complex, but also in sugar allocation into source and sink
cells. The availability of the recently released grapevine genome sequence offers the opportunity to identify
sucrose and monosaccharide transporter gene families in a woody species and to compare them with those of the
herbaceous Arabidopsis thaliana using a phylogenetic analysis.
Results: In grapevine, one of the most economically important fruit crop in the world, it appeared that sucrose
and monosaccharide transporter genes are present in 4 and 59 loci, respectively and that the monosaccharide
transporter family can be divided into 7 subfamilies. Phylogenetic analysis of protein sequences has indicated that
orthologs exist between Vitis and Arabidospis. A search for cis-regulatory elements in the promoter sequences of
the most characterized transporter gene families (sucrose, hexoses and polyols transporters), has revealed that
some of them might probably be regulated by sugars. To profile several genes simultaneously, we created a
macroarray bearing cDNA fragments specific to 20 sugar transporter genes. This macroarray analysis has revealed
that two hexose (VvHT1, VvHT3), one polyol (VvPMT5) and one sucrose (VvSUC27) transporter genes, are highly
expressed in most vegetative organs. The expression of one hexose transporter (VvHT2) and two tonoplastic
monosaccharide transporter (VvTMT1, VvTMT2) genes are regulated during berry development. Finally, three
putative hexose transporter genes show a preferential organ specificity being highly expressed in seeds (VvHT3,
VvHT5), in roots (VvHT2) or in mature leaves (VvHT5).
Conclusions: This study provides an exhaustive survey of sugar transporter genes in Vitis vinifera and revealed that
sugar transporter gene families in this woody plant are strongly comparable to those of herbaceous species.
Dedicated macroarrays have provided a Vitis sugar transporter genes expression profiling, which will likely
contribute to understand their physiological functions in plant and berry development. The present results might
also have a significant impact on our knowledge on plant sugar transporters.

* Correspondence: maryse.laloi@univ-poitiers.fr
† Contributed equally
1
UMR-CNRS-UP 6503 - LACCO - Laboratoire de Catalyse en Chimie
Organique - Equipe Physiologie Moléculaire du Transport de Sucres Université de Poitiers - Bâtiment Botanique - 40 Avenue du Recteur Pineau,
86022 Poitiers cedex, France
Full list of author information is available at the end of the article
© 2010 Afoufa-Bastien et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative
Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited.

Afoufa-Bastien et al. BMC Plant Biology 2010, 10:245
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Background
In plants, sugars (sucrose, monosaccharides, polyols) are
important molecules that constitute not only metabolites
but also nutrients, osmotic and signal molecules. In
numerous species, sucrose is the most prevalent sugar
produced in photosynthetic organs (source) and transported via the phloem over long distances to heterotrophic organs (sink), which depend on a constant supply
of carbohydrates [1]. In sink organs, sucrose is either
directly imported or cleaved by cell wall-bound invertases
into monosaccharides (glucose and fructose), that can be
taken up by the sink cells [2]. In some species, sugar alcohols (polyols), such as mannitol, sorbitol and galactinol
can also be transported on top of sucrose for long-distance
carbon partitioning [3]. In addition to this long-distance
transport, sugars can also be allocated in the different
organelles of source and sink cells, and more and more
biochemical and molecular studies argue for the transport
of hexoses into the chloroplast [4] the vacuoles [5], and
the Golgi apparatus [6]. Therefore, it is now clearly established that not only the loading and the unloading of the
conducting complex, but also the allocation of sugars into
source and sink cells is controlled by sugar transporters
mediating the transport of sucrose [7-9], reducing monosaccharides [10], or polyols [11-13]. Since the cloning of
the first monosaccharide transporter in Chlorella [14], the
first sucrose transporter in Spinacia oleracea [15], and the
first polyol transporter in Apium graveolens [11], many
genes belonging to these families have been isolated from
various species. The complete Arabidopsis genome has
been described to contain 9 sucrose transporter-like
sequences [8] and a monosaccharide transporter(-like)
gene family, including 53 members grouped into 7 subfamilies [10]. Furthermore, the evolutionary analysis of plant
monosaccharide transporters revealed that these seven
subfamilies are ancient in land plants [16].
Despite the progress made in identifying genes encoding
sugar transporters, little is known about the transcriptional
regulation of these genes. Arabidopsis microarray data
(Genevestigator: https://www.genevestigator.com; The
BAR: http://bbc.botany.utoronto.ca) and some plant transporter gene expression patterns have indicated that developmental and environmental factors could regulate the
expression of sugar transporters. Furthermore, evidence is
provided that the expression of some sugar transporter
genes is regulated by sugars as described for sugar transporter genes in yeast [17], for VvHT1, a grapevine hexose
transporter [18-20] and for sucrose transporter genes from
rice, OsSUT1 [21] and sugar beet, BvSUT1 [22-24]. All
these data suggest that the expression of sugar transporters might be regulated at the transcriptional level by distinct but usually converging signalling pathways,
depending on either developmental and environmental

Page 2 of 22

cues or metabolic and hormonal signals. In spite of the
evidence for the role of sugar signalling in the transcriptional control of some transporter genes, the in silico analysis of promoter regions of different genes involved in
carbon metabolism, sugar storage, mobilization and transport clearly demonstrates the absence of common sugar
specific cis-elements [25-27]. This analysis is consistent
with the fact that in plants, several types of transcription
factors (bZIP, WRKY, AP2, MYB, B3, EIN3) are required
for sugar signalling and are involved in sugar-regulation of
gene expression [27,28]. Considering that the analysis of
sugar transporter orthologs in different species might help
to better understand their biological function, we analyzed
the recently sequenced Vitis vinifera genotype PN40024
[29] in order to identify sugar transporter gene families in
this species. This work will represent the first exhaustive
analysis for sugar transporters in ligneous plant as most of
the already known sugar transporters have been characterized from herbaceous species. In woody plants, only 4
sucrose transporters have been already described in Vitis
[30-32] (GenBank: AF439321), 2 in Citrus sinensis (GenBank: AY098891, AY098894), 2 in Hevea brasiliensis
(GenBank: ABJ51934, ABK60189) and one in Juglans regia
[33,34]. Seven hexose transporters in Vitis [35-37], 2 in
Juglans regia [34] and few polyol transporters in Prunus
cerasus [38], in Malus domestica [39] and in Olea europea
[40] were also reported. Furthermore during the last decade, Vitis vinifera has become an interesting model to
study fruit maturation. It is now clearly established that
the onset of ripening (veraison) is characterized by an
important accumulation of glucose and fructose in
vacuoles of the mesocarp cells [41]. In grapevine, sucrose
is the main carbohydrate used for long distance transport
and after reaching the phloem of the berry, it is unloaded
into the apoplast, possibly cleaved by apoplastic invertases,
and sucrose or hexoses can than be transported into the
mesocarp. In the cytoplasm of the mesocarp cells, sucrose
and hexoses must be transported into the vacuole via
tonoplastic transporters. The identification and the characterization of sugar transporter genes in Vitis vinifera are
therefore important steps in understanding the roles of
these proteins in grapevine development as well as in
grape ripening process and may further highlight our
knowledge on plant sugar transporters.
The present study reports on the identification of
sucrose and monosaccharide-like transporter genes in
the Vitis vinifera genome, on their phylogenetic analysis
in comparison with Arabidopsis transporters, on their
promoter sequences analysis. The construction of specialized cDNA macroarrays used to determine the
expression pattern for 20 of these genes in grapevine
vegetative organs and during berry ripening is also
described.

Afoufa-Bastien et al. BMC Plant Biology 2010, 10:245
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Figure 1 Maximum likelihood phylogeny of Vitis vinifera sugar transporter proteins. The tree was produced using MUSCLE and PhyML
with the JTT amino acid substitution model, a discrete gamma model with 4 categories and an estimated shape parameter of 1.213.
Bootstrapping was performed with 100 replicates. For accession numbers of Vitis sugar transporter sequences see Additional file 1. Vitis ORFs
names were simplified, Vv indicating GSVIVT000.

Afoufa-Bastien et al. BMC Plant Biology 2010, 10:245
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Results
Identification of sugar transporters from Vitis vinifera

Blastp searches of the grapevine genome proteome 8×
database, using the amino acid sequences of the 9
sucrose transporters and the 53 monosaccharide transporters from A. thaliana as query, allowed the identification of 65 ORFs encoding putative sugar transporters
in V. vinifera (Additional file 1). Among these ORFs,
only 4 encode previously described sucrose transporters
[30-32] (GenBank: AF439321) and no additional one
could be identified. The 61 other ORFs seem to encode
putative monosaccharide transporters (MST). Phylogenetic analysis of the 65 V. vinifera identified protein
sequences using the maximum likelihood (ML) method
(Figure 1) reveals that sucrose and monosaccharide
transporters form two separate groups. Furthermore in
agreement with the phylogeny observed for A. thaliana
MST [10,16], 7 distinct subfamilies (I-VII) could be
clearly identified in the Vitis monosaccharide transporter group (Additional file 1 and Figure 1).
Vitis vinifera Sucrose Transporters (VvSUC, VvSUT)

The 4 amino acids sequences encoding the already
described sucrose transporters named VvSUC11/
VvSUT1, VvSUC12, VvSUC27 and VvSUT2 [30-32]
share 40 to 59% similarity between each other and fall
into three sucrose transporter subgroups already
described (Figure 2) [7-9]. VvSUT2 and VvSUC27
shows 59% similarity and belong to the dicots specific
SUT1 subfamily including high affinity sucrose transporters exhibiting apparent K m value between 0.07 and

Figure 2 Maximum likelihood phylogeny of Vitis vinifera and
Arabidopsis thaliana sucrose transporter proteins. The tree was
produced using MUSCLE and PhyML with the JTT amino acid
substitution model, a discrete gamma model with 4 categories and
an estimated shape parameter of 0.872. Bootstrapping was
performed with 100 replicates. Accession numbers for Arabidopsis
thaliana transporters are: At1g71880 (AtSUC1), At1g22710 (AtSUC2),
At2g02860 (AtSUC3), At1g09960 (AtSUC4), At1g71890 (AtSUC5),
At5g43610 (AtSUC6), At1g66570 (AtSUC7), At2g14670 (AtSUC8),
At5g06170 (AtSUC9); for Vitis ones see Additional file 1.

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2 mM. However, VvSUC27 has been described to be a
low-affinity/high-capacity sucrose transporter showing a
Km value for sucrose ranged between 8.0 and 10.5 mM
[42]. The structure of VvSUT2 and VvSUC27 genes is
quite similar, both being around 2380 bp long and containing 4 exons separated by 3 introns (Additional file
1). VvSUC12 shows 66.6% similarity with AtSUC3 and
presents an extended domain at the N terminus and an
elongated central cytoplasmic loop; two structural characteristics specific to the SUT2/SUC3 subfamily [43].
Furthermore, VvSUC12 gene is a very long gene (more
than 10 kb) containing 14 exons interrupted by 13
introns (Additional file 1); such exon/intron organization is also described for AtSUC3. The K m value for
sucrose (1.36 mM) reported for this transporter [32]
seems, however, higher than that described for other
members of this subfamily, showing either a low affinity
(AtSUC3: Km = 11.7 mM) or no sucrose transport function. Finally, although VvSUC11 has a high affinity for
sucrose (Km = 0.88 mM; [32]) it shows 67.9% similarity
with AtSUC4 and falls into the SUT4 subfamily including all low-affinity plant sucrose transporters with Km
value ranging between 5 mM and 6 mM.
Vitis vinifera putative Hexose Transporters (VvHT;
subfamily I)

Among the identified ORFs, 22 showed high similarity (40
to 82%) with the AtSTP (Sugar Transport Protein) subfamily members. Among these, 5 correspond to the already
well known V. vinifera hexose transporters named VvHT
(Vitis vinifera hexose transporter) such as VvHT1, VvHT2,
VvHT3 (also named VvHT7), VvHT4 and VvHT5 [35-37],
(Additional file 1). Therefore, the 17 newly identified ORFs
were named VvHT8 to VvHT24. VvHT8 amino acid
sequence shows 99.4% similarity with VvHT1 and the
main differences between the two nucleotide sequences
reside in some single nucleotide polymorphism and in the
length of a microsatellite sequence in the 3’UTR region.
Considering that the chromosomal location of VvHT1 and
VvHT8 is not determined, it is difficult to conclude if these
sequences represent two independent genes, two alleles of
the same gene or possibly one single gene. VvHT9 and
VvHT10 share 98.5% similarity between each other and
around 73% with VvHT11. Interestingly, the three corresponding genes are located in a tandem repeat region, on
chromosome 14. In a similar way, VvHT14 to VvHT24
form a cluster on chromosome 13 and the 11 corresponding amino acid sequences show very high similarity (more
than 90%). VvHT20 and VvHT21, which are located
nearby on chromosome 13, contain the two first exons and
the last exon of a monosaccharide transporter, respectively.
A detailed amino acid sequence analysis revealed that these
two partial ORFs are wrongly annotated and might constitute a single monosaccharide transporter, in the third exon

Afoufa-Bastien et al. BMC Plant Biology 2010, 10:245
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of which a stop codon (TAG) replaces a tryptophan residue (TGG). It is therefore tempting to suggest that this
point mutation at the origin of the false annotation, might
be due to a sequencing error, but we can not exclude that
it could be real. Finally VvHT22, VvHT23 and VvHT24
are partial MST, whose sequences do not seem to be fully
sequenced, missing either the N-ter or the C-ter region, or
both. Therefore, considering that VvHT8 might be identical to VvHT1 and that VvHT20 and VvHT21 are probably
a single protein, we can estimate that the grape genome
might contain 20 putative hexose transporters. In this
VvHT subfamily, the exon-intron organization seems to be
conserved as all completely sequenced genes contain 4
exons separated by 3 introns with the exception of VvHT4
and VvHT5. Phylogenetic analysis (Figure 1) reveals that
the VvHT subfamily seems to be divided into two subclades, at the basis of which is located VvHT5 and
VvHT12 both present on chromosome 5. VvHT12 is
located at the basis of a subclade having a bootstrap value
of 100 and containing the 10 closely related transporters
from chromosome 13 and 9 (VvHT14 to VvHT24).
Furthermore, if we exclude VvHT1 and VvHT8, 3 sisterpairs with a strong bootstrap support (≥ 84%) could be
identified, VvHT3/VvHT13 (56.8% similarity); VvHT9/
VvHT10 (98.5% similarity) and VvHT2/VvHT4 (52.4%
similarity). Finally, phylogenetic analysis of A. thaliana and
V. vinifera sugar transporter proteins allowed us to identify
six ortholog pairs between both species (Figure 3) such
as VvHT1/AtSTP1 (81.9%), VvHT2/AtSTP5 (65.4%),
VvHT3/AtSTP7 (77.2%), VvHT4/AtSTP3 (60.8%), VvHT5/
AtSTP13 (82%), VvHT13/AtSTP14 (75.6%). Five of these
pairs are supported by bootstrap value of 100%.
Vitis vinifera putative Tonoplast Monosaccharide
Transporters (VvTMT; subfamily II)

We have also identified three ORFs, which show the
strongest similarity (58.3 to 72%) to the 3 A. thaliana
Tonoplast Monosaccharide Transporters (AtTMT;
[44]). All three Vitis ORFs show an extended middle
loop between the putative trans-membrane helices
six and seven in a similar way as the AtTMT. GSVIVT
00002919001 is identical to a V. vinifera sugar transporter already mentioned in the literature and called
VvHT6 [36]. Considering that it shows a higher similarity with AtTMT (58.8 to 70.9%) than with the
VvHT proteins (15 to 26.3%), we renamed it VvTMT1.
Similarly, GSVIVT00036283001 and GSVIVT000
19321001 were called VvTMT2 and VvTMT3, respectively. The exon-intron organization seems to be conserved in the three genes as they contain all 5 exons
separated by 4 introns (Additional file 1). Phylogenetic
tree performed with Vitis sugar transporter amino acid
sequences (Figure 1) reveals that the 3 VvTMT form a
clade, which is closely related to putative myo-inositol

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transporters (VvINT) and vacuolar glucose transporters
(VvVGT) (described below). Furthermore, phylogenetic
analysis using Vitis and Arabidopsis sequences
(Figure 4) confirms that TMT sequences from both
species form a single clade, with a strong bootstrap
support (100%), but within which low bootstrap values
(≤ 45%) indicate unresolved nodes and fail to detect
sister-pairs between both species.
Vitis vinifera putative Polyol/Monosaccharide Transporters
(VvPMT; subfamily III)

Five ORFs show highest similarity (41.4 to 72.1%) with
the 6 A. thaliana polyol transporters and have been
therefore named VvPMT1 to VvPMT5. V. vinifera putative polyol transporter amino acids sequences share 40%
to 76.8% similarity between themselves and the corresponding genes present all the same structure with 2
exons separated by a single intron. Phylogenetic analysis
performed with the A. thaliana and V. vinifera polyol
transporters (Figure 4) reveals that VvPMT1 and
VvPMT4 form with AtPMT4 a separated clade.
VvPMT2 is at the basis of a second clade, which can be
divided into two groups, one including VvPMT3,
AtPMT3 and AtPMT6 and the second AtPMT1,
AtPMT2, AtPMT5 and VvPMT5. Only AtPMT4 and
VvPMT4 could be identified as putative orthologs.
Vitis vinifera putative ERD6-like Transporters (subfamily
IV)

Twenty-two ORFs showing strongest similarity with the
19 AtERD6-like proteins were identified (Additional file
1) and share 36.2 to 93.2% similarity with each other.
Among them, 6 ORFs correspond to partial sequences
in which either the beginning or the end of the protein
are not clearly identified. However after a more precise
sequence analysis, we were able to realize the full annotation for GSVIVT00006084001 and GSVIVT0000
6097001. Fourteen ORFs are located on chromosome
14, in a region of tandem gene duplications, three other
ORFs are carried by chromosome 5 and two partial
ORFs by chromosome 12. The 22 ERD6-like proteins
fall into the same subfamily supported by a strong bootstrap value (99%) and 14 loci formed 7 sister pairs
(Figure 1). The phylogenetic analysis of the amino acid
sequences of ERD6-like transporters from A. thaliana
and V. vinifera (Figure 5) reveals that these transporters
can be classified into 4 major groups. A first group
includes 7 AtERD6-like located on the 5 chromosomes
of A. thaliana and 9 VvERD6-like located on 3 chromosomes (5, 7, 14) of V. vinifera. A second small group
including transporters from both species (At1g19450,
At1g75220, Vv14605001 and Vv249200001) was also
identified. Inversely, the two last groups include protein
sequences from only one species. The Arabidopsis

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Figure 3 Maximum likelihood phylogeny of Vitis vinifera and Arabidopsis thaliana hexose transporter proteins. The tree was produced
using MUSCLE and PhyML with the JTT amino acid substitution model, a discrete gamma model with 4 categories and an estimated shape
parameter of 1.025. Bootstrapping was performed with 100 replicates. Accession numbers for Arabidopsis thaliana transporters are: At1g11260
(AtSPT1), At1g07340 (AtSTP2), At5g61520 (AtSTP3), At3g19930 (AtSTP4), At1g34580 (AtSTP5), At3g05960 (AtSTP6), At4g02050 (AtSTP7), At5g26250
(AtSTP8), At1g50310 (AtSTP9), At3g19940 (AtSTP10), At5g23270 (AtSTP11), At4g21480 (AtSTP12), At5g26340 (AtSTP13), At1g77210 (AtSTP14); for
Vitis ones see Additional file 1.

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Figure 4 Maximum likelihood phylogeny of Vitis vinifera and Arabidopsis thaliana monosaccharide transporter proteins. The tree was
produced using MUSCLE and PhyML with the JTT amino acid substitution model, a discrete gamma model with 4 categories and an estimated
shape parameter of 1.448. Bootstrapping was performed with 100 replicates. Accession numbers for Arabidopsis thaliana transporters are:
At1g20840 (AtTMT1), At4g35300 (AtTMT2), At3g51490 (AtTMT3), At2g43330 (AtINT1), At1g30220 (AtINT2), At2g35740 (AtINT3), At4g16480 (AtINT4),
At3g03090 (AtVGT1), At5g17010 (AtVGT2), At5g59250 (AtVGT3), At2g16120 (AtPMT1), At2g16130 (AtPMT2), At2g18480 (AtPMT3), At2g20780
(AtPMT4), At3g18830 (AtPMT5), At4g36670 (AtPMT6); for Vitis ones see Additional file 1. Vitis ORFs names were simplified, Vv indicating
GSVIVT000.

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Figure 5 Maximum likelihood phylogeny of Vitis vinifera and Arabidopsis thaliana ERD6-like transporter proteins. The tree was
produced using MUSCLE and PhyML with the JTT amino acid substitution model, a discrete gamma model with 4 categories and an estimated
shape parameter of 1.404. Bootstrapping was performed with 100 replicates. Arabidopsis transporters are indicated with complete ORFs names,
for Vitis ones ORFs names were simplified, Vv indicating GSVIVT000.

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Table 1 Common putative cis-acting elements identified in the VvSUC/VvSUT, VvHT, VvTMT and VvPMT promoter
sequences
Cis-element name

Sequence

Response

ARR1AT

NGATT

Cytokinines

Maximum number of copies/promoter
36

CIACADIANLELHC

CAANNNNATC

Circadian expression

4**

DOFCOREZM
EBOXBNNAPA

AAAG
CANNTG

C-metabolism, leaf
Light, ABA, seeds

40
36

EECCRCAH1

GANTTNC

CO2-responsive

7

GATABOX

GATA

Light, leaf, shoot

29

GT1CONSENSUS

GRWAAW

Light, leaf, shoot

39

GTGANTG10

GTGA

Pollen

19

IBOXCORE

GATAA

Light, leaf, shoot

13

MYBST1

GGATA

Myb trans activator

4

MYCCONSENSUSAT
PYRIMIDINEBOXOSRAMY1A

CANNTG
CCTTTT

ABA, abiotic stress
Sugar repression, seeds

36
9*

POLLEN1LELAT52

AGAAA

Pollen

28

RAV1AAT

CAACA

Root, rosette leaves

9

ROOTMOTIFTAPOX1

ATATT

Root

45

SEF4MOTIFGM7S

RTTTTTR

Seed, storage protein

23

WBOXATNPR1

TTGAC

Desease resistance

8*

WBOXHVISO1

TGACT

Sugar, SUSIBA2

9

WBOXNTERF3
WRKY71OS

TGACY
TGAC

Wounding, ERF3
GA repressor, ABA

16
25

Promoter sequence analysis was performed via PLACE. Cis-element name, sequence motifs and signalling pathway are presented. Up to copies/promoter
indicates the highest number of cis-acting element found in one promoter. * indicates that the motif is found in all promoters except VvPMT2. ** indicates that
the motif is found in all promoters except VvPMT2, VvTMT3 and VvPMT2.

group contains 10 proteins located on three different
chromosomes (1, 3, 5) including ERD6 [45,46], SFP1
and SFP2 [47]. The Vitis group includes 11 proteins,
9 of which are encoded by genes located on chromosome 14.
Vitis vinifera putative Vacuolar Glucose Transporters
(VvVGT; subfamily V)

Two Vitis ORFs, named VvVGT1 and VvVGT2, show
the highest similarity with the 3 AtVGT (Vacuolar
Glucose Transporter)-like transporters. In Arabidopsis
AtVGT1 and AtVGT2 have been shown to be localized
in the tonoplast and glucose transport activity has been

demonstrated for AtVGT1 [48]. On the contrary
AtVGT3 is postulated to be localized in chloroplast
membrane as this protein presents a N-terminal extension carrying a potential signal for plastid targeting. Phylogenetic tree (Figure 4) indicates clearly that VvVGT1
is the closest to AtVGT1 and AtVGT2 and that
VvVGT2, which presents a N-terminal extension, is
more closely related to AtVGT3.
Vitis vinifera putative Inositol Transporters (VvINT;
subfamily VI)

We identified 3 ORFs showing the strongest similarity
with the 4 AtINT (Inositol transporter) already described

Table 2 Unique cis-acting elements identified only in the promoter sequence of a single sugar transporter gene
Cis-element name

Sequence

Response

Copy number

Gene

ABREZMRAB28

CCACGTGG

CRTDREHVCBF2

GTCGAC

Drought, ABA

2

HT5

Cold, drought

2

GARE2OSREP1

TAACGTA

GA, germination

HT5

1

TMT3

GBOX10NT

GCCACGTGCC

Leaf, root, flower, pollen

1

HT2

GBOXLERNCS
LREBOXIIPCCHS1

MCACGTGGC
TCCACGTGGC

Light, overlap ABA
Cold, drought, ABA

1
1

HT5
HT5

1 to 3

VvSUC/VvSUT

MYBCOREATCYCB1

AACGG

cell cycle, cyclin

NONAMERMOTIFATH3H4

CATCCAACG

meristem,

1

TMT3

ZDNAFORMINGATCAB1

ATACGTGT

Light, leaf, shoot

1

SUC11

Promoter sequence analysis was performed via PLACE. Cis-element names, sequence motifs, signalling pathways and the number of copies for each element are
presented. Gene indicates the corresponding gene in which the cis-elements are found.

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in Arabidopsis. To our knowledge, only two AtINT have
been already characterized. AtINT4 is described as a
high-affinity, plasma membrane-localized H+/symporter
specific for myo-inositol [49]. AtINT1 is a tonoplast-localized H+/inositol symporter that mediates the efflux of
inositol that is generated during the degradation of inositol-containing compounds in the vacuolar lumen [50].
The three Vitis ORFs were named VvINT1-3 according
to their highest similarity with AtINT (Figure 4).
Other Vitis vinifera putative monosaccharide Transporters
(VvpGlcT/VvSGB1; subfamily VII)

Finally, 4 ORFs show high similarity with the members
of the Arabidopsis AtpGlcT/AtSGB1 subfamily, which
includes proteins showing homology with a putative glucose transporter (pGlcT) of the chloroplast inner envelope membrane from spinach [4] and with a Golgilocalized hexose transporter homolog (suppressor of G
protein beta1:SGB1; [6]). The ORF GSVIVT000
38247001 is identical to a V. vinifera sugar transporter
already mentioned in the literature and called VvpGlT
[20,36]. Phylogenetic tree (Figure 4) reveals that inside
this subfamily, the proteins separate into 3 groups having strong bootstrap support (100%). VvpGlT and
AtpGlcT fall into the same group, which includes also
SopGlcT from spinach (not shown). This observation
can argue in favor of a chloroplastic localization of
VvpGlT even if the precise localization of this transporter is not demonstrated. In a similar way, the fact that
Vv16716001 and Vv34389001 form a second group with
At1g67300 and SGB1 indicates that these two Vitis
putative transporters could be localized in Golgi apparatus. Finally Vv25939001 forms a third group with
At1g05030.
Search for cis-elements putatively involved in the
transcriptional regulation of sugar transporter genes

We have identified a 2 kb promoter region for each of
the 29 fully sequenced genes from the four mostly studied sugar transporter families: VvSUC/SUT, VvHT,
VvTMT and VvPMT (Additional file 2). For only three
genes VvHT14 (1455 bp), VvTMT3 (1619 bp) and
VvPMT2 (623 bp), the identified sequence is shorter
due to the presence of an other ORF located less than
2kb upstream of these transporter genes. A PLACE analysis has been applied to these promoter sequences and
the 216 identified cis-acting elements have been classified per sugar transporter subfamily, for comparison.
Cis-elements common to all promoters

In a first approach, 20 common cis-regulatory elements
conserved in the promoter regions of the 29 analyzed
sequences have been identified (Table 1). Only the shortest promoter VvPMT2 (623pb) is missing three of these

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elements, namely CIACADIANLELHC, PYRIMIDINEBOXOSRAMY1A and WBOXATNPR1. Moreover, these
common consensus sequences are highly repetitive
displaying up to 45 copies into a 2 kb promoter. This
might be due to the their limited size (4 to 7 bases), and
to their high variability (1, 2 and 3 degenerated nucleotides per motif of 5, 6 and 7 bases, respectively). These
common cis-acting elements are able to confer expression in distinct plant organs, such as leaves, shoots,
roots, seeds, and flowers (pollen). They are also responsive to different plant hormones (abscisic acid, gibberellins, ethylen, cytokinins), as well as to several
environmental factors (light, CO 2 , biotic and abiotic
stresses). At least a quarter of these common consensus
sequences (EBOXBNNAPA, GATABOX, GT1CONSENSUS, GTGANTG10, IBOXCORE) are required for
the transcriptional regulation by light, and this mainly
in leaves and shoots. This is in agreement with the roles
of the studied transporters in sugar allocation between
source- and sink-organs. Finally, the presence of the box
CIACADIANLELHC, absent only in the VvPMT2,
VvTMT3 and VvHT12 promoters, strongly suggests the
importance of circadian regulation for sugar transporter
gene expression.
Cis-elements present only in a single promoter

A second complementary approach was targeted to
unique consensus sequences present in the promoter of
only one sugar transporter gene, thereby implying some
expression specificity. The few unique identified cis-elements (Table 2) are characterized by longer sequences
(5 to 10 bases), and usually lacking any nucleotide
variability. Interestingly, among the 9 gene specific
motifs identified, 4 are present only in the VvHT5 promoter, 2 in VvTMT3, one in VvHT2 and another one in
VvSUC11. This means that a limited number of gene
specific cis-acting elements is concentrated in the promoter regions of few sugar transporter genes. For example the hexose transporter gene VvHT5 is the only one
among the 29 genes studied, displaying 4 unique motifs
(ABREZMRAB28, CRTDREHVCBF2, GBOXLERNCS,
LREBOXIIPCCHS1) in its promoter. Finally, another
specific cis-element strongly restricted to VvSUC/
VvSUT genes is the motif MYBCOREATCYCB1 (Table
2), required for transcriptional regulation of cyclin B1 at
two different phases of the cell cycle, G1/S and G2/M
transitions [51].
Cis-elements involved in sugar regulated transcription

We have studied the transcriptional regulation of sugar
transporter genes through the repertory of the main
promoter motifs potentially involved in sugar-regulated
transcription, and this in combination with other metabolic and hormonal signalling. Additional file 3 summarizes the careful comparison of the following

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Figure 6 Macroarray analysis of VvSUC/VvSUT (A), VvHT (B), VvTMT (C) and VvPMT (D) genes expression in grapevine vegetative
organs. RNA was isolated from vegetative organs collected from 20 independent plants. Gene transcript levels were normalized against four
reference genes (GAPDH, EF1a, EF1g, actin). Each value represents the mean of six replicates obtained with two independent experiments. Red
point indicates an expression value higher than the mean of the expression value for all genes in the tested organ (mean value of relative
expression for young leaf: 0.03; mature leaf: 0.05; petiole: 0.07; stem: 0.05; root: 0.05; tendril: 0.07).

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Figure 7 RNA gel blot analysis of V. vinifera sugar transporter genes transcript levels in vegetative organs. RNA was isolated from
vegetative organs collected from 20 independent plants. Gene transcript levels were quantified using Image Quant 5.2 software and normalized
against GAPDH gene expression. Red point indicates an expression value higher than the mean of the expression value for all genes in the
tested organ (mean value of relative expression for young leaf: 1.65; mature leaf: 5.62; petiole: 6.01; stem: 3.55; root: 3.37; tendril: 4.10).

consensus sequences: i) elements for sugar responsiveness as the SURE boxes [52], the bipartite sucrose box 3
[53], the CGACGOSAMY3 [54], the CMSRE [55], the
SP8 and WBOXHVISO1 sequences enabling the binding
of some WRKY-type proteins at the example of SPF and
SUSIBA2 [56-58]; ii) sequences common for hormonal
and metabolic (sugar) signals perception as the S-box
for sugar and ABA [59], the MYBGAHV for gibberellins
(GAs) induction and sugar repression [60]; the GARC
complex consisting of the AMYBOX1 and 2 [61], and
PYRIMIDINE boxes for GAs, ABA and sugar regulation.
There is at least one gene for each subfamily displaying the majority of chosen sugar responsive motifs
(VvHT1 and VvHT8 - 9 motifs, VvSUC11 and VvSUT2 8 motifs, VvHT5, VvTMT3 and VvPMT5 - 7 motifs),
thus suggesting a possible transcriptional control dependent on sugars as metabolic signals (Additional file 3).
The sucrose box 3, is the most frequently found cisacting element, present at 1 to 4 copies in all studied
promoters except VvHT15, VvTMT1 and VvPMT2. On
the contrary, the CMSRE1IBSPOA element, involved in
sucrose positive regulation is only found in promoter

regions of VvHT2, VvHT5 and VvTMT3. The sucrose
transporter gene family, is the only one displaying the
SURE2 motif in the promoter regions of VvSUC11 and
VvSUC27. The sugar responsiveness CGACGOSAMY3
box is carried only by VvHT genes (VvHT1, VvHT3,
VvHT5, VvHT8, VvHT11), and not by the other subfamilies. Similarly, the S-box (CACCTCCA) usually closely associated to the light-responsive G-box, is carried
also only by VvHT genes, namely VvHT1, VvHT8 and
VvHT11. Inversely, the motif MYBGAHV involved in
sugar and GA signalling pathways, is displayed by
VvSUC/SUT, VvPMT and VvTMT genes, but is lacking
in VvHT ones. Finally it appears that VvHT1 and
VvHT8 promoter sequences are the only one to contain
a putative GARC complex. A more detailed comparison
into the VvHT subfamily reveals that VvHT1 and
VvHT8 promoters are carrying the same cis-elements
(with the exception of one more copy of the PYRIMIDINEBOXHVEPB1 for VvHT1). This indicates that as the
coding sequences, the promoter regions for these two
putative genes present also a very high similarity
(96.7%). This argues in favor of the assumption that

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Figure 8 Macroarray analysis of VvSUC/VvSUT (A), VvHT (B), VvPMT (C), VvTMT (D) genes expression during grapevine berry and seed
development. For each developmental stages, RNA was isolated from all berries collected from 5 independent grapes. Gene transcript levels
were normalized against four reference genes (GAPDH, EF1a, EF1g, actin). Each value represents the mean of six replicates obtained with two
independent experiments. Red point indicates an expression value higher than the mean of the expression value for all genes in the tested
organ (mean value of relative expression for berry 2 WAF: 0.056; berry 10 WAF: 0.046; berry 11 WAF: 0.033; berry 13 WAF: 0.029; seed 10 WAF:
0.032; seed 11 WAF: 0.058).

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these are either alleles of the same gene or represent the
same gene, as already suggested in this study. Such consideration is valuable for another couple of genes mentioned above, VvHT9 and VvHT10, sharing a strong
sequence similarity, displaying the same cis-acting elements in their promoter region, and carried on the
same chromosome. A third gene VvHT11 is present in
tandem repeat with both VvHT9 and VvHT10, by the
same chromosome, thus suggesting that they may be
products of successive duplications.
V. vinifera sugar transporter genes expression in
vegetative organs

In order to study the expression pattern of grapevine
sugar transporter genes identified above and belonging
to the VvSUC/VvSUT, VvHT, VvTMT, and VvPMT subfamilies, we have developed sugar transporter macroarray membranes. Specific regions for each sugar
transporter (Additional file 4) have been identified in
the 3’UTR of the corresponding nucleotide sequences,
amplified by PCR using Chardonnay genomic DNA and
spotted on nylon membrane. VvHT8, VvHT9, VvHT10
and VvHT14 to VvHT24, could not be considered in
this expression analysis, as it was not possible to found
specific DNA region for these transporters, due to their
high sequence similarity either with VvHT1 or between
each other. To determine the gene expression patterns
in vegetative organs, macroarrays were hybridized with
33
P-labelled first-strand cDNA synthesized from total
RNA isolated from young leaves, mature leaves, petioles,
stems, roots and tendrils from 10 weeks-old grapevine
plants grown under aeroponic conditions. These culture
conditions have been used in order to collect all main
vegetative organs at the same development stage and in
the same conditions and allowed an easy access to the
root system without damage. Among the sucrose transporters only VvSUC27 is detected at a high level in
petioles, stems and tendrils, its transcripts being less
abundant in young leaves, mature leaves and roots (Figure 6A). VvSUC11 and VvSUC12 are detected in all
organs but at a weaker level and VvSUT2 is the less
expressed sucrose transporter being only weakly
detected in young leaves and roots. Concerning the
VvHT family (Figure 6B), 3 genes (VvHT1, VvHT3 and
VvHT11) are expressed in all the tested organs at a relatively high level (expression value higher than the mean
of expression value for all genes) with VvHT1 and
VvHT11 being less expressed in young leaves and tendrils, respectively. VvHT2 and VvHT5 seem to present a
more specific expression. VvHT2 is expressed at a
higher level in roots than in the other organs and
VvHT5 is highly expressed in mature leaves and presents a weaker expression in roots and young leaves.
The three other hexose transporters (VvHT4, VvHT12,

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VvHT13) are weakly detected, indicating a low expression in the tested organs. The three VvTMT are also
detected at a very low level in all organs (Figure 6C).
Among the polyol/monosaccharide transporters (Figure
6D), only VvPMT5 could be significantly detected in the
six organs. It shows a strong expression in mature
leaves, petioles and tendrils, and a weaker expression in
stems, roots and young leaves. The other VvPMT are
weakly expressed in all the tested organs.
In order to validate the results obtained with macroarray
hybridizations and to confirm the expression pattern of
the sugar transporter genes, we performed a Northern
blot analysis for few genes. The results presented in
Figure 7 clearly show similar expression patterns for
most of the tested genes. VvHT1 shows the weakest
expression in young leaves. VvHT2 seems to be specifically expressed in roots. VvHT5 presents a weak expression in all organs, except in mature leaves. VvTMT1
shows a global weak expression in all organs. VvPMT5
is highly expressed in mature leaves, petioles and tendrils. Finally, VvSUC27 shows high amount of transcripts in petioles, stems and tendrils. Few discrepancies
were however observed. First, the expression level
detected for VvHT3 is the highest in mature leaves
when detected by Northern blot which is not the case
using macroarray. Second, VvHT3 and VvHT11 show a
higher expression in macroarray than in Northern blot
analysis. For VvHT11 signals obtained with Northern
blot were too low to be correctly quantified. Furthermore, we could confirm using both methods that
VvHT3 is expressed at a higher level than VvHT2,
VvHT4, VvHT5, and VvHT11 in almost all organs.
Taken together, all these results indicate that few transporter genes (VvHT1, VvHT3, VvPMT5, VvSUC27) are
the most expressed in almost all vegetative organs and
that VvHT2 and VvHT5 are more specifically expressed
in roots and mature leaves, respectively.
Sugar transporter genes expression during grape berry
development

In order to study the expression of sugar transporter
genes during berry development, we further hybridized
the sugar transporters macroarray membranes with
33
P-labelled first-strand cDNA synthesized from total
RNA isolated from berries and seeds. Four developmental
stages for berries - fruit set (2WAF), veraison (10WAF),
ripening (11WAF), ripe berries (13WAF) - and two for
seeds (10 and 11 WAF) were used. Among the sucrose
transporter genes, VvSUC11 and VvSUC12 are both
expressed during berry development at a similar level to
that detected in vegetative organs (Figure 8A). The weakest expression for these two genes is observed in berries
at the stage of fruit set. On the contrary, VvSUC27, the
most expressed sucrose transporter gene in vegetative

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organs is less expressed than VvSUC11 and VvSUC12 in
berries while VvSUT2 is weakly or not detected. Three
hexose transporters are expressed during berries development (Figure 8B). VvHT2 is expressed during the 4 tested
stages and presents a maximum at the veraison and during ripening. Inversely, VvHT3 and VvHT11 are
expressed at an equivalent level during the 4 developmental stages. VvHT1, VvHT4, VvHT5, VvHT12 and
VvHT13 are poorly or not detected at any stage. The
expression of two putative tonoplast monosaccharide
transporters (VvTMT1 and VvTMT2) which is weak at
the fruit set increases significantly at the veraison (figure
8C). Furthermore, the expression level of these two genes
is higher in berries at the veraison and during ripening
than in vegetative organs. On the contrary, VvTMT3
does not seem to be expressed in berries at any stage of
development. Polyols transporters are not highly
expressed in berries, only VvPMT1 is weakly detected
during the 4 stages and VvPMT5 is expressed mainly at
the fruit set stage (Figure 8D). Macroarray hybridization
performed with first-strand cDNA synthesized from total
RNA isolated from seeds reveals clearly that the expression of VvHT3 and VvHT5 is very high in seeds and
increases during seed development (Figure 8B). Two
other transporter genes VvSUC12 and VvSUC27 were
also expressed in seeds but at a lower level, and their
expression stays stable during the two tested developmental stages (Figure 8A).

Discussion
Phylogenetic analysis of Vitis vinifera sugar transporter
genes

The search for sugar transporters in the Vitis vinifera
translated genome has identified 4 sucrose and 59 putative
monosaccharide transporters including 20 VvHT (Hexose
Transporters), 3 VvTMT (Tonoplastic Monosaccharide
Transporters), 5 VvPMT (Polyol/Monosaccharide Transporters), 3 VvINT (INositol Transporter), 2 VvVGT
(Vacuolar Glucose Transporters), 4 pGlT/SGB1 and 22
ERD6-like transporters. As expected, phylogenetic analysis
performed with these sugar transporter proteins revealed
that sucrose and monosaccharide transporters form two
distinct groups (Figure 1). This analysis allowed us to identify only 4 Vitis sucrose transporters, which confirms that,
as all other analyzed plants, Vitis possesses a small sucrose
transporter gene family, in which one gene (VvSUC12)
belongs to the SUT2 subfamily. Interestingly, in Vitis as in
Arabidopsis, the VvHT and the ERD6-like form the largest
multigenic subfamilies. In Vitis, this may be due to the
presence of 4 repeated regions, encompassing VvHT and
ERD6-like genes. Two duplicated regions located on chromosomes 13 and 14 contain 9 and 3 VvHT, respectively.
The 2 other regions carried by chromosomes 5 and 14 display respectively 3 and 14 ERD6-like genes. Similarly, in

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Arabidopsis, the large expansion of AtSTP subfamily has
been correlated with 3 segmental duplications and one
tandem duplication as well as the expansion of the
AtERD6-like subfamily by 2 segmental duplications and 6
tandem duplications [16]. Furthermore, based on significant differences in size observed in the STP and ERD6like subfamilies, between the non-vascular (moss) and the
vascular (gymnosperm and angiosperm) lineages, it has
been suggested that the expansion of these two subfamilies
could be related to the evolution of vascular plants. This is
reflecting the increased importance of the sugar transport
and sugar transporters in vascular plants [16]. In agreement with this hypothesis, the AtERD6/VvERD6-like phylogenetic tree (Figure 5) clearly shows that ERD6
transporters from both species fall into four different
groups, two of them containing either AtERD6 or
VvERD6 transporters only. This indicates that in both species, the expansion of the ERD6-like subfamilies has
occurred quite recently, after the separation of these two
species.
Sugar-responsive elements in sugar transporter gene
promoters suggest their regulation by sugars

The in silico search for cis-acting elements reveals several common and highly repetitive motifs in sugar transporter gene promoters. These cis-acting elements such
as DOF (DNA-binging with one finger) proteins, may
play a role not only in the regulation of sugar transporter gene expression in terms of activity level, but also
plausibly in terms of response specificity via a combinatory control. Such a control has already been suggested
for AtSUC2 [62] the expression of which in the companion cell is regulated by the close cooperation of binding sites for a DOF and a putative HD-Zip transcription
factors. Several transporter gene promoters display an
important concentration of sugar-responsive elements
suggesting their possible transcriptional regulation by
sugars. To our knowledge, the transcriptional regulation
of VvHT1 by glucose is the only one to be clearly
demonstrated [18-20] and this is confirmed by the fact
that VvHT1 promoter contains the highest number of
sugar responsive motifs. This highlights the power of
the in silico analysis as a first step toward the functional
characterization of promoter regions. Finally, the MYBCOREATCYCB1 sequence exclusively found in SUC/
SUT promoters is not surprising in regard to the
sucrose-dependent induction of Cyclin D3 gene expression [63], thus suggesting a possible concomitant regulation of some sucrose transporter genes in the cell cycle.
Sucrose transporter genes expression in Vitis vinifera

The expression patterns detected for the sucrose transporter genes, using macroarrays, are in good agreement
with those described in the literature with two main

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exceptions: the absence of high expression of VvSUC11
in seeds and of VvSUC27 in roots, as reported by [30].
On the contrary, we confirmed that, in berries,
VvSUC11 and VvSUC12 transcripts are present at all
developmental stages and accumulate slightly at the
onset of ripening [30,31]. VvSUC11 is closely related to
AtSUC4, which has been localized in the tonoplast of A.
thaliana mesophyll cells [64]. Furthermore, VvSUC12
falls into the SUT2/SUC3 group which contains very
low affinity sucrose transporters for which different
putative physiological functions have been proposed
including their putative involvement in sucrose import
into several sink tissues [43,65,66]. Therefore, it is
tempting to suggest that VvSUC12 is probably involved
either in phloem unloading or in sucrose import into
berry tissues and that VvSUC11 might be responsible
for sucrose accumulation in berry vacuoles. This
hypothesis will have to be verified with the precise localization of these two transporters. We noticed that
VvSUC27 is the most expressed sucrose transporter
gene in vegetative organs and that its expression is relatively low in berries. Considering its high expression in
petioles, stems and tendrils and the fact that it is closely
related to members of the SUT1 subfamily, VvSUC27 is
probably responsible for phloem loading and sugar
retrieval during long-distance transport. Finally, the
weak expression level observed for the less characterized
Vitis transporter gene VvSUT2 makes it difficult to
assign a specific role for this transporter.
Hexose transporter genes expression in Vitis vinifera

The present phylogenetic analysis indicates that VvHT1
shows highest similarity with AtSTP1. Both are high
affinity glucose transporters showing Km value of 70 μM
[36] and 20 μM [67], respectively. During the last decade, different authors have reported various expression
patterns for VvHT1 such as a strong expression in berries and young leaves [35], a preferential expression in
sink organs [18], an expression in conducting bundle of
leaves, petioles and berries [36] or an expression
increasing with leaf development [37]. During berry
development, VvHT1 expression was described to show
two peaks (one at the time of anthesis, the other after
veraison) [35] or to decline rapidly during ripening
[37,68]. This second expression pattern was supported
by the detection of VvHT1 protein only in young green
berries [20]. Our results (Figures 6 and 8) clearly confirm that VvHT1 belongs to the hexose transporters that
are poorly expressed in berries, but is one of the mostly
expressed VvHT in vegetative organs including leaves,
petioles, stems, roots and tendrils. Furthermore, its
expression increases during leaf development.
VvHT2 shows the highest similarity with AtSTP5
which has not been yet characterized. Different reports

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describing the expression of VvHT2 have shown that
VvHT2 is weakly expressed in leaves whatever the stage
of development [35] and that the transcript level is high
in young berries and declines around veraison
[35,37,68]. Our data (Figures 6 and 8) confirm not only
VvHT2 expression in leaves and during berry development, but indicate also its weak expression in almost all
vegetative organs except for roots in which it seems to
be strongly expressed.
VvHT3 has been described to be one of the mostly
expressed VvHT in leaves, with increasing expression
during leaf development. In young berries, its expression
is high, decreases around veraison and increases again
around the phase of sugar storage [37]. The expression
pattern determined in our experiment (Figures 6 and 7)
correlates with that described previously. However, we
found that this transporter, which is expressed in all
vegetative organs is also highly expressed in seeds (Figure 8), in which its expression seems to increase during
development. Interestingly, VvHT3 shows the highest
similarity with AtSTP7 for which a strong seed expression is also suggested by microarray hybridization data
(genevestigator, BAR). Although the localisation and the
functionality of these transporters are unknown, they
might have a determinant function in sugar storage in
seeds and/or in embryo development.
VvHT4 is poorly expressed in all the tested organs and
hardly detectable in berries (Figures 6 and 8), in accordance with previous report describing a very weak
expression in berry and leaf development [37]. VvHT4
has been characterized as a glucose transporter showing
a high affinity for glucose (Km : 137 μM), higher than
that reported for AtSTP3 (K m = 2 mM) the closest
related Arabidopsis transporter. A physiological role,
either to support wounded tissue or in the retrieval of
monosaccharides released during cell damage and cell
wall degradation, has been proposed for AtSTP3, based
on its induction after wounding [2]. A more precise
characterization of VvHT4 is therefore required to verify
if the expression of this gene is also regulated after
wounding or in response to other stresses.
VvHT5 was found to be less expressed than VvHT1
and VvHT3 in developing leaves and during berry development [37]. Our experiments confirmed that VvHT5
transcripts are hardly detected in berries, however they
are predominant in seeds, at least for the two tested
developmental stages (Figure 8). VvHT5 shows the highest similarity with AtSTP13 and both have similar high
affinity for glucose (K m = 89 μM and K m = 74 μM,
respectively) [37,69]. Furthermore, the expression of
these two transporters is described to be induced in
response to pathogen attack [69,70]. This indicates that
these genes could be involved in pathogen starvation
and/or in a sugar signalling pathway in plant defense.

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Figure 9 Schematic representation of preferential expression of Vitis vinifera sugar transporter genes in the different vegetative
organs and during berry development. This summary is based on the expression data described in the present report. For each organ, only
genes with an expression value higher than the mean of the expression value for all genes are presented. Genes indicated in bold are the most
expressed in the indicated organ. Underlined genes are those showing a preferential expression or being induced during the development of
the considered organ.

The presence of four unique cis-acting elements
(Table 2) and of a cluster of three ABRE motifs [70] in
the promoter region of VvHT5 gene is in agreement
with the regulation of its expression by ABA and biotic
stress but also suggests its involvement in abiotic stress
responses.
Three putative hexose transporters named VvHT11,
VvHT12 and VvHT13, that have never been described
earlier, have been identified. Our phylogenetic analysis
(Figure 3) revealed that VvHT11 and VvHT12 are each
located at the basis of one of the two duplicated regions
involved in the expansion of the VvHT subfamily.
Furthermore, this analysis allowed us to identify
AtSTP14 and VvHT13 as orthologs, but no orthologs for
VvHT11 and VvHT12 could be found. The expression
pattern of VvHT11, the weak expression of VvHT12 and
VvHT13 in all vegetative organs and in berries are not

sufficient to suggest putative physiological functions for
these three transporters. However recent data indicates
that VvHT13 is induced by necrotrophic fungi and could
be involved as VvHT5 in biotic stress response (AfoufaBastien et al., personal communication).
Putative tonoplastic monosaccharide transporter genes
expression in Vitis vinifera

Three putative tonoplastic monosaccharide transporters were identified and named VvTMT1-3. Expression
data indicate that, even if the expression of VvTMT1
and VvTMT2 is present in all the tested vegetative
organs, it is highest in developing berries. The expression of VvTMT1 is described to increase during berry
development [71] with a maximum near the start of
veraison [68]. Our data not only confirm this expression pattern but suggest also that at least two

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transporters, VvTMT1 and VvTMT2, might probably
play a significant role during ripening. Although their
cellular localisation and their transport activity have
not been determined so far, they could be involved in
hexose accumulation into vacuoles of berry cells.
Furthermore, considering that the expression of
AtTMT1 and AtTMT2 has been reported to be
induced by drought, salt, and cold treatments [44], it
would be interesting to verify, if the expression of the
VvTMT genes is also regulated under stress conditions,
particularly in vegetative organs, where they are weakly
expressed in normal conditions.
Polyols transporter genes expression in Vitis vinifera

Since 2001, many polyol transporters have been identified and characterized in sorbitol or mannitol-translocating plants, where they are described to be responsible
for the loading of polyols into the phloem and their
transfer to sink organs [11,12,38-40]. More recently,
polyol transporters have been identified and characterized in non-polyol-translocating species such as A. thaliana [72-74] which contains 6 polyol transporters, the
physiological role of which is still unknown. In Vitis, the
expression of only one EST encoding a putative PMT
has been already briefly mentioned in the literature and
was shown to be weakly expressed during berry development [68]. Our in silico analysis, indicates that the Vitis
genome contains 5 putative polyol transporter genes.
Among them only one named VvPMT5 was highly
expressed in vegetative organs and only at the fruit set.
However as grapevine has not been described as a species transporting polyols in the phloem, the role of this
transporter is far from being clear.

Conclusions
The present work represents the first exhaustive analysis
of sugar transporter genes in a woody plant. The identification of grapevine sugar transporter genes and their comparative analysis with the Arabidopsis ones has indicated a
strong conservation of these genes between herbaceous
and woody plants as well as some expansions of particular
functional subfamilies such as hexoses and ERD6-like
transporters. In this paper, we developed macroarrays to
profile the expression of 20 of these transporters simultaneously in different organs. Our results not only confirmed some expression data already described in the
literature but also demonstrated that four sugar transporter genes are expressed in almost all vegetative organs
(VvHT3, VvHT11, VvPMT5, VvSUC27), few transporters
are more specifically expressed in roots (VvHT2), mature
leaves (VvHT5) and/or in seeds (VvHT3, VvHT5) and
three others are regulated during berry development
(VvHT2, VvTMT1, VvTMT2) (Figure 9). The present
results might help to elucidate the biological function of

Page 18 of 22

sugar transporters in V. vinifera development particularly
during berry ripening and would also have a significant
impact on our knowledge on plant sugar transporters in
general. The in silico analysis of promoter sequences
revealed the presence of cis-regulatory elements involved
in sugar signalling, and represents a first step towards the
understanding of the regulation of sugar transporter gene
expression via metabolic, hormonal and environmental
signals. More and more evidences suggest that sugar
transporter genes are regulated under various conditions.
Thus, the macroarray analysis described in this paper
could constitute a powerful approach to investigate the
sugar transporter response to environmental factors in
grapevine.

Methods
Identification of sugar transporter genes in V. vinifera
genome

V. vinifera sugar transporter genes were identified performing a Blastp analysis [75] against the V. vinifera
proteome 8X database, on Genoscope website http://
www.genoscope.cns.fr/externe/GenomeBrowser/Vitis
using each A. thaliana monosaccharide and sucrose
transporter amino acid sequences as query, and an
E-value of 1,00E-04 as threshold. Furthermore, the 2 kb
region upstream of the start codon for each gene was
considered as the promoter sequence.
Sequence similarities, phylogeny and promoter sequence
analysis

Sequence similarities were determined performing Clustal V multiple alignments using Lasergene software
(DNASTAR, USA). Phylogenetic analysis of V. vinifera
and A. thaliana sugar transporter protein sequences
was performed using maximum likelihood and the
http://www.phylogeny.fr website[76]. For this, protein
sequences alignment was performed using the MUSCLE
program [77], and maximum likelihood trees with 100
bootstrap replicates were constructed with the PHYML
program [78,79] and the JTT amino acid substitution
model. Phylogenic tree was visualized using Treedyn
program [80]. Search for cis-regulatory elements in promoter sequences was performed using the PLAnt Cis
acting regulatory DNA Elements database (PLACE:
http://www.dna.affrc.go.jp/PLACE/index.html)
Plant material and growth conditions

Vitis vinifera cv. Chardonnay plants were cultivated
in vitro, for 7 weeks (46 days), on McCown Woody Plant
Medium (Duchefa, The Netherlands), pH 5.8, supplemented with 20 g.l-1 sucrose, with 16 h photoperiod at
24°C. Plants were then transferred to an aeroponic culture system and grown with Gibeaut solution [81] under
controlled conditions (16 h photoperiod, 23°C; 70% RH

Afoufa-Bastien et al. BMC Plant Biology 2010, 10:245
http://www.biomedcentral.com/1471-2229/10/245

day/18°C; 65% RH night). After 24 days, young and
mature leaves, stems, roots, petioles and tendrils were
sampled, immediately frozen in liquid nitrogen and
stored at -80°C. V. vinifera cv. Chardonnay berries were
harvested in the 2007 growing season (between 25th June
and 10 th September) from grapevines grown in SRPV
Poitou-Charentes fields (Biard, Poitiers, France). Berries
were sampled at 2, 10, 11 and 13 weeks after flowering
(WAF) corresponding to fruit set, veraison, and two
maturation developmental stages, the last one being 10
days before harvest. After freezing in liquid nitrogen,
seedless berries and seeds were stored at -80°C.
RNA isolation

Total RNA was isolated from grapevine tissues as previously described by Valtaud and coworkers (2009) [82].
For macroarray analysis, RNA was treated with RNasefree DNaseI (QIAGEN, Germany) in order to eliminate
contaminant DNA and purified using the RNeasy Mini
Kit (QIAGEN, Germany), according to the RNA clean
up protocol.
Cloning of specific cDNA fragments and macroarray
membrane spotting

Specific DNA regions for each sugar transporter and
reference genes (actin, EF1a, EF1g and GAPDH) have
been identified in the 3’UTR of the nucleotide sequence
and amplified by PCR using Chardonnay genomic DNA
and specific primers (Additional file 4). PCR products
were purified with Wizard®SV Gel and Clean-Up System
(Promega, USA) according to manufacturer’s protocol,
cloned into the pGEM®-T Easy Vector (Promega, USA)
and sequenced using the ABI PRISM® BigDye® Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems,
USA).
Specific cDNA fragments have been amplified from
the obtained plasmids by PCR using one specific primer
and T7 primer. For each reaction, 1 μl of plasmid DNA
solution was used as template in a 50 μl PCR reaction,
containing 1× Green GoTaq® Flexi Buffer, 2 mM
MgCl2, 0.4 μM of each primer, 0.2 mM of each deoxynucleotide and 1.25 U of GoTaq® DNA Polymerase
(Promega, USA). Amplification reactions included an
initial denaturation step at 94°C for 5 min, followed by
30 cycles of 1 min at 94°C, 1 min at 52°C, 1 min at 72°
C and a final extension of 5 min at 72°C. All PCR products were purified using the Wizard®SV Gel and
Clean-Up System (Promega, USA) according to manufacturer’s protocol.
Each cDNA fragment was dotted in triplicate on a 6×
SSC-soaked nylon Hybond™-N+ membrane (GE Healthcare, UK), using a 96-well Bio-Dot® Microfiltration
Apparatus (BIO-RAD, Canada). The amount of cDNA

Page 19 of 22

per spot was 50 ng for sugar transporter and 100 ng of
each reference gene. Three dots of 50 ng of salmon
sperm DNA were used as internal negative control.
Membranes were then incubated in a denaturing solution (1.5 M NaCl, 0.5 M NaOH), in a neutralizing solution (1.5 M NaCl, 0.5 M TRIS-HCl pH 8), and finally
washed in 2× SSC solution. DNA was cross-linked to
the membrane by exposure to UV light (120 mJ/cm 2)
using a crosslinker (Bio-Link-BLX-E254).
Macroarray membrane hybridization

For the synthesis of 33P-labeled cDNA, 30 μg of DNase
treated total RNA were retro-transcribed using 2 μM
oligo d(T) 16 , 0.5 mM dATP, dTTP, dGTP, 2.26 μM
dCTP, 0.33 μM [a-33P]-dCTP (10mCi ml-1) and 800 U
of M-MLV Reverse Transcriptase (Promega, USA).
Labeled products were then treated with 10 U of Ribonuclease H (Promega, USA) and purified on illustra™
Probe Quant™ Micro Columns (GE Healthcare, UK).
Prehybridization and hybridization were carried out at
65°C using Church solution (1% BSA, 1 mM EDTA,
0.25 M Na2HPO4-NaH2PO4 and 7% SDS). After 16-20 h
of hybridization, membranes were washed twice in 2×
SSC; 0.1% SDS for 15 min, twice in 1× SSC; 0.1% SDS
for 15 min at 65°C and exposed in a Storage Phosphor
Cassette for 48 h and images were acquired using a
Typhoon TRIO Imager (GE Healthcare, UK). Spot finding, quantification and background subtraction were
done with ImageQuant TL 7.0 program (GE Healthcare,
UK). Spots were considered as present only if higher
than the mean of salmon sperm negative control and
then normalized using the mean of 4 reference genes
(actin, EF1a, EF1g and GAPDH).
Northern blotting

Total RNA (20 μg) isolated from different organs of
grapevine plants were separated by electrophoresis in
denaturing formaldehyde 1.2% agarose gel and then
transferred to Hybond™-N Nylon membrane (GE
Healthcare, UK). DNA probes designed on 3’UTR
regions of genes of interest were produced by PCR reaction and labeled with [a- 32 P]-dCTP using Prime-aGene® Labelling System (Promega, USA) according to
manufacturer’s protocol. Prehybridization and hybridization were performed as described for macroarray. Membranes were washed in 2× SSC, 0.1% SDS for 15 min, in
1× SSC, 0.1% SDS for 15 min and in 0.5× SSC, 0.1%
SDS for 5 min at 65°C. Membranes were exposed for 48
h in a Storage Phosphor Cassette and scanned as performed for macroarray analysis. Quantification and
background correction were done using Image Quant
5.2 software. Reported signals were then normalized to
GAPDH expression value.

Afoufa-Bastien et al. BMC Plant Biology 2010, 10:245
http://www.biomedcentral.com/1471-2229/10/245

Additional material
Additional file 1: Sucrose and Monosaccharide transporter genes
identified in Vitis vinifera genome. Vitis proteome 8× ID, attributed
name, chromosomal position, gene length, number of introns and exons,
ORF length, protein length, estimated protein molecular weight and pI,
GenBank ID and reference are indicates when available. Genes written in
italics are partial.
Additional file 2: Sucrose and Monosaccharide transporter genes
promoter sequences identified in Vitis vinifera genome. Vitis
proteome 8× ID, attributed name, chromosomal position and promoter
length are indicated.
Additional file 3: Cis-acting elements potentially involved in sugarregulated transcription identified in the VvSUC/VvSUT, VvHT, VvTMT
and VvPMT promoter sequences. Promoter sequence analysis was
performed via PLACE. Cis-element names, sequence motifs, signalling
pathways and the number of copies for each element are presented for
each promoter. x(xx): number of motif types (total number of identified
motifs). i) Elements for sugar responsiveness. ii) Elements for sugar and
hormonal signals perception.
Additional file 4: Oligonucleotide primers used to amplify specific
3’UTR of sugar transporter and reference genes. Gene name,
oligonucleotide sequences and length of the amplified cDNA fragments
are indicated.

Page 20 of 22

5.

6.

7.
8.
9.
10.
11.

12.

13.

14.
Acknowledgements
The authors thank Mathieu Célérier and Emilie Sohier for their contribution
in retrieving gene and promoter sequences. We are grateful to Freddy
MANCEAU from SRPV (service regional de la protection des végétaux)
Poitou-Charentes in BIARD (FRANCE) for Chardonnay berries gift.
Author details
1
UMR-CNRS-UP 6503 - LACCO - Laboratoire de Catalyse en Chimie
Organique - Equipe Physiologie Moléculaire du Transport de Sucres Université de Poitiers - Bâtiment Botanique - 40 Avenue du Recteur Pineau,
86022 Poitiers cedex, France. 2UMR Génétique et Horticulture (GenHort) INRA/INH/UA - BP 60057 - 49071 Beaucouzé cedex, France.

15.

16.

17.
18.

19.

Authors’ contributions
DAB and AM were equally involved in plant culture, RNA isolation, cDNA
cloning, macroarrays and Northern blot hybridizations, data analysis and
participated in genome search and manuscript writing. JJ participated in
genome search, cDNA cloning and macroarrays perfecting. PCT helped in
macroarrays perfecting and was involved in revising the manuscript critically
for intellectual content. RL was involved in revising the manuscript critically
for intellectual content and gave final approval of the version to be
published. RA performed cis-elements identification and wrote the
corresponding part of the manuscript. ML conceived the study, carried out
the genome search, the gene and promoter sequences analysis, the
phylogenetic analysis, participated in building macroarrays and wrote the
manuscript. All authors read and approved the final manuscript.

24.

Received: 16 June 2010 Accepted: 12 November 2010
Published: 12 November 2010

25.

References
1. Lemoine R: Sucrose transporters in plants: update on function and
structure. Biochim Biophys Acta 2000, 1465(1-2):246-262.
2. Buttner M, Sauer N: Monosaccharide transporters in plants: structure,
function and physiology. Biochim Biophys Acta 2000, 1465(1-2):263-274.
3. Zimmermann MH, Ziegler : List of sugars and sugar alcohols in sieve-tube
exudates. In Encyclopedia of Plant Physiology. Edited by: Zimmermann MH,
Milbum JA. Berlin: Springer Verlag; 1975:480-503.
4. Weber A, Servaites JC, Geiger DR, Kofler H, Hille D, Groner F, Hebbeker U,
Flugge UI: Identification, purification, and molecular cloning of a
putative plastidic glucose translocator. Plant Cell 2000, 12(5):787-802.

20.

21.

22.
23.

26.

27.
28.
29.

Martinoia E, Massonneau A, Frangne N: Transport processes of solutes
across the vacuolar membrane of higher plants. Plant Cell Physiol 2000,
41(11):1175-1186.
Wang HX, Weerasinghe RR, Perdue TD, Cakmakci NG, Taylor JP, Marzluff WF,
Jones AM: A Golgi-localized hexose transporter is involved in
heterotrimeric G protein-mediated early development in Arabidopsis.
Mol Biol Cell 2006, 17(10):4257-4269.
Kuhn C: A comparison of the sucrose transporter systems of different
plant species. Plant Biol 2003, 5:215-232.
Sauer N: Molecular physiology of higher plant sucrose transporters. FEBS
Lett 2007, 581(12):2309-2317.
Kuhn C, Grof CP: Sucrose transporters of higher plants. Curr Opin Plant
Biol 2010, 13:1-11.
Buttner M: The monosaccharide transporter(-like) gene family in
Arabidopsis. FEBS Lett 2007, 581(12):2318-2324.
Noiraud N, Maurousset L, Lemoine R: Identification of a mannitol
transporter, AgMaT1, in celery phloem. Plant Cell 2001,
13(3):695-705.
Ramsperger-Gleixner M, Geiger D, Hedrich R, Sauer N: Differential
expression of sucrose transporter and polyol transporter genes during
maturation of common plantain companion cells. Plant Physiol 2004,
134(1):147-160.
Juchaux-Cachau M, Landouar-Arsivaud L, Pichaut JP, Campion C,
Porcheron B, Jeauffre J, Noiraud-Romy N, Simoneau P, Maurousset L,
Lemoine R: Characterization of AgMaT2, a plasma membrane mannitol
transporter from celery, expressed in phloem cells, including phloem
parenchyma cells. Plant Physiol 2007, 145(1):62-74.
Sauer N, Tanner W: The hexose carrier from Chlorella. cDNA cloning of a
eucaryotic H+-cotransporter. FEBS Lett 1989, 259(1):43-46.
Riesmeier JW, Willmitzer L, Frommer WB: Isolation and characterization of
a sucrose carrier cDNA from spinach by functional expression in yeast.
Embo J 1992, 11(13):4705-4713.
Johnson DA, Hill JP, Thomas MA: The monosaccharide transporter gene
family in land plants is ancient and shows differential subfamily
expression and expansion across lineages. BMC Evol Biol 2006, 6:64.
Rolland F, Winderickx J, Thevelein JM: Glucose-sensing and -signalling
mechanisms in yeast. FEMS Yeast Res 2002, 2(2):183-201.
Atanassova R, Leterrier M, Gaillard C, Agasse A, Sagot E, Coutos-Thevenot P,
Delrot S: Sugar-regulated expression of a putative hexose transport gene
in grape. Plant Physiol 2003, 131(1):326-334.
Cakir B, Agasse A, Gaillard C, Saumonneau A, Delrot S, Atanassova R: A
grape ASR protein involved in sugar and abscisic acid signaling. Plant
Cell 2003, 15(9):2165-2180.
Conde C, Agasse A, Glissant D, Tavares R, Geros H, Delrot S: Pathways of
glucose regulation of monosaccharide transport in grape cells. Plant
Physiol 2006, 141(4):1563-1577.
Matsukura C, Saitoh T, Hirose T, Ohsugi R, Perata P, Yamaguchi J: Sugar
uptake and transport in rice embryo. Expression of companion cellspecific sucrose transporter (OsSUT1) induced by sugar and light. Plant
Physiol 2000, 124(1):85-93.
Chiou TJ, Bush DR: Sucrose is a signal molecule in assimilate partitioning.
Proc Natl Acad Sci USA 1998, 95(8):4784-4788.
Vaughn MW, Harrington GN, Bush DR: Sucrose-mediated transcriptional
regulation of sucrose symporter activity in the phloem. Proc Natl Acad
Sci USA 2002, 99(16):10876-10880.
Ransom-Hodgkins WD, Vaughn MW, Bush DR: Protein phosphorylation
plays a key role in sucrose-mediated transcriptional regulation of a
phloem-specific proton-sucrose symporter. Planta 2003, 217(3):483-489.
Sheen J, Zhou L, Jang JC: Sugars as signaling molecules. Curr Opin Plant
Biol 1999, 2(5):410-418.
Delrot S, Atanassova R, Maurousset L: Regulation of sugar, amino acid and
peptide plant membrane transporters. Biochim Biophys Acta 2000,
1465(1-2):281-306.
Rolland F, Baena-Gonzalez E, Sheen J: Sugar sensing and signaling in plants:
conserved and novel mechanisms. Annu Rev Plant Biol 2006, 57:675-709.
Ramon M, Rolland F, Sheen J: Sugar Sensing and Signaling. The
Arabidopsis Book Rockville: American Society of Plant Biologists; 2008.
Jaillon O, Aury JM, Noel B, Policriti A, Clepet C, Casagrande A, Choisne N,
Aubourg S, Vitulo N, Jubin C, et al: The grapevine genome sequence
suggests ancestral hexaploidization in major angiosperm phyla. Nature
2007, 449(7161):463-467.

Afoufa-Bastien et al. BMC Plant Biology 2010, 10:245
http://www.biomedcentral.com/1471-2229/10/245

30. Davies C, Wolf T, Robinson SP: Three putative sucrose transporters are
differentially expressed in grapevine tissues. Plant Sci 1999, 147:93-100.
31. Ageorges A, Issaly N, Picaud S, Delrot S, Romieu C: Identification and
functional expression in yeast of a grape berry sucrose carrier. Plant
Physiol Biochem 2000, 38(3):177-185.
32. Manning K, Davies C, Bowen HC, White PJ: Functional characterization of
two ripening-related sucrose transporters from grape berries. Ann
Botlondon 2001, 87:125-129.
33. Decourteix M, Alves G, Brunel N, Ameglio T, Guillio A, Lemoine R, Petel G,
Sakr S: JrSUT1, a putative xylem sucrose transporter, could mediate
sucrose influx into xylem parenchyma cells and be up-regulated by
freeze-thaw cycles over the autumn-winter period in walnut tree
(Juglans regia L.). Plant Cell Environ 2006, 29(1):36-47.
34. Decourteix M, Alves G, Bonhomme M, Peuch M, Ben Baaziz K, Brunel N,
Guilliot A, Rageau R, Ameglio T, Petel G, et al: Sucrose (JrSUT1) and hexose
(JrHT1 and JrHT2) transporters in walnut xylem parenchyma cells: their
potential role in early events of growth resumption. Tree Physiol 2008,
28(2):215-224.
35. Fillion L, Ageorges A, Picaud S, Coutos-Thevenot P, Lemoine R, Romieu C,
Delrot S: Cloning and expression of a hexose transporter gene expressed
during the ripening of grape berry. Plant Physiol 1999, 120(4):1083-1094.
36. Vignault C, Vachaud M, Cakir B, Glissant D, Dedaldechamp F, Buttner M,
Atanassova R, Fleurat-Lessard P, Lemoine R, Delrot S: VvHT1 encodes a
monosaccharide transporter expressed in the conducting complex of
the grape berry phloem. J Exp Bot 2005, 56(415):1409-1418.
37. Hayes MA, Davies C, Dry IB: Isolation, functional characterization, and
expression analysis of grapevine (Vitis vinifera L.) hexose transporters:
differential roles in sink and source tissues. J Exp Bot 2007,
58(8):1985-1997.
38. Gao Z, Maurousset L, Lemoine R, Yoo SD, van Nocker S, Loescher W:
Cloning, expression, and characterization of sorbitol transporters from
developing sour cherry fruit and leaf sink tissues. Plant Physiol 2003,
131(4):1566-1575.
39. Watari J, Kobae Y, Yamaki S, Yamada K, Toyofuku K, Tabuchi T, Shiratake K:
Identification of sorbitol transporters expressed in the phloem of apple
source leaves. Plant Cell Physiol 2004, 45(8):1032-1041.
40. Conde C, Silva P, Agasse A, Lemoine R, Delrot S, Tavares R, Geros H:
Utilization and transport of mannitol in Olea europaea and implications
for salt stress tolerance. Plant Cell Physiol 2007, 48(1):42-53.
41. Davies C, Robinson SP: Sugar accumulation in grape berries. Cloning of
two putative vacuolar invertase cDNAs and their expression in
grapevine tissues. Plant Physiol 1996, 111(1):275-283.
42. Zhang YL, Meng Q, Zhu HL, Guo Y, Gao HY, Luo YB, Jiang L: Functional
characterization of a LAHC sucrose transporter isolated from grape
berries in yeast. Plant Growth Regul 2008, 54:71-79.
43. Meyer S, Lauterbach C, Niedermeier M, Barth I, Sjolund RD, Sauer N:
Wounding enhances expression of AtSUC3, a sucrose transporter from
Arabidopsis sieve elements and sink tissues. Plant Physiol 2004,
134(2):684-693.
44. Wormit A, Trentmann O, Feifer I, Lohr C, Tjaden J, Meyer S, Schmidt U,
Martinoia E, Neuhaus HE: Molecular identification and physiological
characterization of a novel monosaccharide transporter from
Arabidopsis involved in vacuolar sugar transport. Plant Cell 2006,
18(12):3476-3490.
45. Kiyosue T, Yamaguchi-Shinozaki K, Shinozaki K: Cloning of cDNAs for
genes that are early-responsive to dehydration stress (ERDs) in
Arabidopsis thaliana L.: identification of three ERDs as HSP cognate
genes. Plant Mol Biol 1994, 25(5):791-798.
46. Kiyosue T, Abe H, Yamaguchi-Shinozaki K, Shinozaki K: ERD6, a cDNA clone
for an early dehydration-induced gene of Arabidopsis, encodes a
putative sugar transporter. Biochim Biophys Acta 1998, 1370(2):187-191.
47. Quirino BF, Reiter WD, Amasino RD: One of two tandem Arabidopsis
genes homologous to monosaccharide transporters is senescenceassociated. Plant Mol Biol 2001, 46(4):447-457.
48. Aluri S, Buttner M: Identification and functional expression of the
Arabidopsis thaliana vacuolar glucose transporter 1 and its role in seed
germination and flowering. Proc Natl Acad Sci USA 2007, 104(7):2537-2542.
49. Schneider S, Schneidereit A, Konrad KR, Hajirezaei MR, Gramann M,
Hedrich R, Sauer N: Arabidopsis INOSITOL TRANSPORTER4 mediates highaffinity H+ symport of myoinositol across the plasma membrane. Plant
Physiol 2006, 141(2):565-577.

Page 21 of 22

50. Schneider S, Beyhl D, Hedrich R, Sauer N: Functional and physiological
characterization of Arabidopsis INOSITOL TRANSPORTER1, a novel
tonoplast-localized transporter for myo-inositol. Plant Cell 2008,
20(4):1073-1087.
51. Trehin C, Ahn IO, Perennes C, Couteau F, Lalanne E, Bergounioux C:
Cloning of upstream sequences responsible for cell cycle regulation
of the Nicotiana sylvestris CycB1;1 gene. Plant Mol Biol 1997,
35(5):667-672.
52. Grierson C, Du JS, de Torres Zabala M, Beggs K, Smith C, Holdsworth M,
Bevan M: Separate cis sequences and trans factors direct metabolic and
developmental regulation of a potato tuber storage protein gene. Plant
J 1994, 5(6):815-826.
53. Hattori T, Nakagawa S, Nakamura K: High-level expression of tuberous
root storage protein genes of sweet potato in stems of plantlets grown
in vitro on sucrose medium. Plant Mol Biol 1990, 14(4):595-604.
54. Hwang YS, Karrer EE, Thomas BR, Chen L, Rodriguez RL: Three cis-elements
required for rice alpha-amylase Amy3 D expression during sugar
starvation. Plant Mol Biol 1998, 36(3):331-341.
55. Morikami A, Matsunaga R, Tanaka Y, Suzuki S, Mano S, Nakamura K: Two
cis-acting regulatory elements are involved in the sucrose-inducible
expression of the sporamin gene promoter from sweet potato in
transgenic tobacco. Mol Genet Genomics 2005, 272(6):690-699.
56. Ishiguro S, Nakamura K: The nuclear factor SP8BF binds to the 5’upstream regions of three different genes coding for major proteins of
sweet potato tuberous roots. Plant Mol Biol 1992, 18(1):97-108.
57. Ishiguro S, Nakamura K: Characterization of a cDNA encoding a novel
DNA-binding protein, SPF1, that recognizes SP8 sequences in the 5’
upstream regions of genes coding for sporamin and beta-amylase from
sweet potato. Mol Gen Genet 1994, 244(6):563-571.
58. Sun C, Palmqvist S, Olsson H, Boren M, Ahlandsberg S, Jansson C: A novel
WRKY transcription factor, SUSIBA2, participates in sugar signaling in
barley by binding to the sugar-responsive elements of the iso1
promoter. Plant Cell 2003, 15(9):2076-2092.
59. Acevedo-Hernandez GJ, Leon P, Herrera-Estrella LR: Sugar and ABA
responsiveness of a minimal RBCS light-responsive unit is mediated by
direct binding of ABI4. Plant J 2005, 43(4):506-519.
60. Morita A, Umemura T, Kuroyanagi M, Futsuhara Y, Perata P, Yamaguchi J:
Functional dissection of a sugar-repressed alpha-amylase gene (RAmy1
A promoter in rice embryos. FEBS Lett 1998, 423(1):81-85.
61. Huang N, Sutliff TD, Litts JC, Rodriguez RL: Classification and
characterization of the rice alpha-amylase multigene family. Plant Mol
Biol 1990, 14(5):655-668.
62. Schneidereit A, Imlau A, Sauer N: Conserved cis-regulatory elements for
DNA-binding-with-one-finger and homeo-domain-leucine-zipper
transcription factors regulate companion cell-specific expression of the
Arabidopsis thaliana SUCROSE TRANSPORTER 2 gene. Planta 2008,
228(4):651-662.
63. Riou-Khamlichi C, Menges M, Healy JM, Murray JA: Sugar control of the
plant cell cycle: differential regulation of Arabidopsis D-type cyclin gene
expression. Mol Cell Biol 2000, 20(13):4513-4521.
64. Endler A, Meyer S, Schelbert S, Schneider T, Weschke W, Peters SW, Keller F,
Baginsky S, Martinoia E, Schmidt UG: Identification of a vacuolar sucrose
transporter in barley and Arabidopsis mesophyll cells by a tonoplast
proteomic approach. Plant Physiol 2006, 141(1):196-207.
65. Barth I, Meyer S, Sauer N: PmSUC3: characterization of a SUT2/SUC3-type
sucrose transporter from Plantago major. Plant Cell 2003, 15(6):1375-1385.
66. Hackel A, Schauer N, Carrari F, Fernie AR, Grimm B, Kuhn C: Sucrose
transporter LeSUT1 and LeSUT2 inhibition affects tomato fruit
development in different ways. Plant J 2006, 45(2):180-192.
67. Sauer N, Friedlander K, Graml-Wicke U: Primary structure, genomic
organization and heterologous expression of a glucose transporter from
Arabidopsis thaliana. Embo J 1990, 9(10):3045-3050.
68. Deluc LG, Grimplet J, Wheatley MD, Tillett RL, Quilici DR, Osborne C,
Schooley DA, Schlauch KA, Cushman JC, Cramer GR: Transcriptomic and
metabolite analyses of Cabernet Sauvignon grape berry development.
BMC Genomics 2007, 8:429.
69. Norholm MH, Nour-Eldin HH, Brodersen P, Mundy J, Halkier BA: Expression
of the Arabidopsis high-affinity hexose transporter STP13 correlates with
programmed cell death. FEBS Lett 2006, 580(9):2381-2387.
70. Hayes MA, Feechan A, Dry IB: Involvement of abscisic acid in the
coordinated regulation of a stress-inducible hexose transporter (VvHT5)

Afoufa-Bastien et al. BMC Plant Biology 2010, 10:245
http://www.biomedcentral.com/1471-2229/10/245

71.

72.

73.

74.

75.

76.

77.
78.
79.
80.

81.

82.

Page 22 of 22

and a cell wall invertase in grapevine in response to biotrophic fungal
infection. Plant Physiol 2010, 153(1):211-221.
Terrier N, Glissant D, Grimplet J, Barrieu F, Abbal P, Couture C, Ageorges A,
Atanassova R, Leon C, Renaudin JP, et al: Isogene specific oligo arrays
reveal multifaceted changes in gene expression during grape berry
(Vitis vinifera L.) development. Planta 2005, 222(5):832-847.
Reinders A, Panshyshyn JA, Ward JM: Analysis of transport activity of
Arabidopsis sugar alcohol permease homolog AtPLT5. J Biol Chem 2005,
280(2):1594-1602.
Klepek YS, Geiger D, Stadler R, Klebl F, Landouar-Arsivaud L, Lemoine R,
Hedrich R, Sauer N: Arabidopsis POLYOL TRANSPORTER5, a new member
of the monosaccharide transporter-like superfamily, mediates H
+-Symport of numerous substrates, including myo-inositol, glycerol, and
ribose. Plant Cell 2005, 17(1):204-218.
Klepek YS, Volke M, Konrad KR, Wippel K, Hoth S, Hedrich R, Sauer N:
Arabidopsis thaliana POLYOL/MONOSACCHARIDE TRANSPORTERS 1 and
2: fructose and xylitol/H+ symporters in pollen and young xylem cells. J
Exp Bot 2010, 61(2):537-550.
Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W,
Lipman DJ: Gapped BLAST and PSI-BLAST: a new generation of protein
database search programs. Nucleic Acids Res 1997, 25(17):3389-3402.
Dereeper A, Guignon V, Blanc G, Audic S, Buffet S, Chevenet F, Dufayard JF,
Guindon S, Lefort V, Lescot M, et al: Phylogeny.fr: robust phylogenetic
analysis for the non-specialist. Nucleic Acids Res 2008, , 36 Web Server:
W465-469.
Edgar RC: MUSCLE: multiple sequence alignment with high accuracy and
high throughput. Nucleic Acids Res 2004, 32(5):1792-1797.
Guindon S, Gascuel O: A simple, fast, and accurate algorithm to estimate
large phylogenies by maximum likelihood. Syst Biol 2003, 52(5):696-704.
Anisimova M, Gascuel O: Approximate likelihood-ratio test for branches:
A fast, accurate, and powerful alternative. Syst Biol 2006, 55(4):539-552.
Chevenet F, Brun C, Banuls AL, Jacq B, Christen R: TreeDyn: towards
dynamic graphics and annotations for analyses of trees. BMC
Bioinformatics 2006, 7:439.
Gibeaut DM, Hulett J, Cramer GR, Seemann JR: Maximal biomass of
Arabidopsis thaliana using a simple, low-maintenance hydroponic
method and favorable environmental conditions. Plant Physiol 1997,
115(2):317-319.
Valtaud C, Foyer CH, Fleurat-Lessard P, Bourbouloux A: Systemic effects on
leaf glutathione metabolism and defence protein expression caused by
esca infection in grapevines. Func Plant Biol 2009, 36:260-279.

doi:10.1186/1471-2229-10-245
Cite this article as: Afoufa-Bastien et al.: The Vitis vinifera sugar
transporter gene family: phylogenetic overview and macroarray
expression profiling. BMC Plant Biology 2010 10:245.

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