Carbohydrate Research 368 (2013) 2934

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Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres

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Research progresses on the key enzymes involved in sucrose

metabolism in maize
Xiaodong Ren , Junjie Zhang ,
College of Life Science, Sichuan Agriculture University, Yaan 625014, China

a r t i c l e i n f o a b s t r a c t

Article history: Sucrose, as the major product of photosynthesis, is a vital metabolite and signaling molecule in higher
Received 8 September 2012 plants. Three enzymes are responsible for the synthesis, transport, and degradation of sucrose. In this
Received in revised form 19 October 2012 article, the gene structure, expression and regulation, and the physiological functions of the key enzymes
Accepted 20 October 2012
involved in sucrose metabolism in maize are reviewed, moreover, the existing problems of the sucrose
Available online 27 October 2012
metabolism research were discussed in detail, and we present our ideas for future research.
2012 Elsevier Ltd. All rights reserved.
Keywords:
Maize
Sucrose metabolism
Key enzyme
Gene expression
Gene regulation

1. Introduction enzymatic properties.20,21 In this paper, we focus on summarizing

Sucrose, the predominant product of photosynthesis, is the
main form of long-distance transport and distribution in higher
plants in vivo. After synthesis in autotrophic organs (source), su-
crose is transported into heterotrophic organs (sink) via the
phloem, primarily for synthesis of storage molecules or to provide
energy for the plant itself. At the same time, sucrose is the source
of carbon skeletons involved in the synthesis of starch,17 cellu-
lose,8,9 lipids, proteins, and nucleic acids.10 Sucrose is also an
important signal molecule in plants that controls the transcription
and translation of a number of genes.1114 In addition, sucrose par-
ticipates in the abiotic stress tolerance of plants.1517 In conclusion,
sucrose metabolism (including synthesis and degradation) plays
signicant roles in plant growth and development.
The main metabolic pathways of sucrose are well known. Three
key enzymes participate in sucrose synthesis and degradation,
invertase (b-fructofuranosidase, EC 3.2.1.26, Inv), sucrose synthase
(EC 2.4.1.13, SuSy), and sucrose-phosphate synthase (EC 2.3.1.14,
SPS). Invertase and sucrose synthase are primarily involved in su-
crose degradation, while sucrose-phosphate synthase is mainly
responsible for sucrose synthesis.18,19 In recent years, the genes
encoding these enzymes have been cloned from many plants,
and important advances have been made in the study of their
expression and regulation, structure and function, and related
Corresponding authors. Tel.: +86 08352886126.
E-mail address: junjiezh@163.com (J. Zhang).
These authors contributed to this work equally.
the signicant research progress on these three key enzymes and
their genes in maize, which are very important for maize improve-
ment using molecular techniques.
2. Invertase in maize
2.1. Gene structure and enzymatic characteristics of invertase
Invertase has always been considered a key enzyme in carbohy-
drate metabolism because it catalyzes sucrose cleavage to its
monosaccharide constituents, glucose and fructose, in an irrevers-
ible manner.22 In maize, invertase is separated into soluble (vacu-
olar form, mainly distributed in plant vacuoles) and insoluble
forms (cell wall form, bound to the cell wall), which are both gly-
coproteins and have an optimum pH of 5.0.23 To date, there have
been six genes encoding invertases identied in maize, Ivr1,24
Ivr2,25 Incw1,26 Incw2 (Mn1),27 Incw3, and Incw4.28 Ivr1 and Ivr2 en-
code vacuolar forms, while Incw1, Incw2, Incw3, and Incw4 encode
cell wall forms. Ivr1 consists of seven exons/six introns, and its sec-
ond exon is only 9 bp, one of the smallest functional exons cur-
rently known in a plant genome.24 The putative genomic
sequence of Ivr2 includes three exons/two introns, and the rst
exon, rst intron, and second exon regions include polymorphic
sites.29 Incw1 and Incw2 both contain seven exons/six introns,
which is typical of most other acid invertase genes.30 The Incw1
gene encodes two distinct transcripts: Incw1-S (small) and Incw1-
L (large), whose size variation is attributable to different lengths
in the 30 untranslated region.26,31 The Incw3 and Incw4 genes in-

0008-6215/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.carres.2012.10.016
30 X. Ren, J. Zhang / Carbohydrate Research 368 (2013) 2934
clude six exons/ve introns and ve exons/four introns, respec-
tively.28 Alignment of the deduced amino acid sequences of these
genes shows that the two soluble invertases only have 3644%
similarity to the four insoluble invertases, but IVR1 shows 64% se-
quence similarity to IVR2. Among the four insoluble invertases,
INCW1 has the highest sequence similarity to INCW2 (70%), while
both INCW2 and INCW3 show only 46% sequence similarity to
INCW4. To date, there have been many studies on the gene expres-
sion, regulation and physiological functions of invertases in maize.
2.2. Roles of invertase
Cell wall invertase is spatially and temporally the rst enzyme
to metabolize incoming sucrose in the developing seeds of maize.32
It also controls the endosperm sink strength and the developmen-
tal stability of the maternal cells in the pedicel.33 INCW2 is pleio-
tropic in maize since its mutations, miniature1 (mnl), can result
in numerous changes, the most obvious being seeds that show a
loss of nearly 80% of their weight compared to the wild type.33
Additionally, INCW2 expression is temporally coincident with
endosperm cell division.32 LeClere et al. pointed out that invert-
ase-mediated sucrose cleavage directly or indirectly regulates the
levels of key plant hormones during maize seed development.34
One recent study showed that both the endosperm and embryo
mass in mn1 genotypes are reduced signicantly at the early stages
of development, and more importantly, the mn1 endosperm shows
reductions in gene expression, ranging from 70% to 99% of Mn1
plants, for both the sucrose-starch and sucrose-energy pathways.35
These studies suggest that INCW2 affects plant growth and devel-
opment in many respects in maize.
Several lines of evidence have suggested that invertase is asso-
ciated with the fertility of kernels in maize. At high temperatures,
it was found that the aborting kernels had much lower pedicel sol-
uble acid invertase activities than non-aborting kernels, and the
aborting kernels failed to synthesize starch in the endosperm and
elevated the ratio of sucrose to hexoses in the pedicel during early
growth.36 During the critical, abortion-sensitive phase of young
ovary development in maize, drought repression of Ivr2 also pre-
ceded changes in Incw2, and the drought-induced decrease and
developmental changes of ovary hexose to sucrose ratio were cor-
related with the activity of soluble but not insoluble invertase.37
Later, some studies demonstrated that all ovary genes for sucrose
metabolic enzymes were inhibited at low water potential (ww),
but a gene for an invertase inhibitor that appeared to be constitu-
tively expressed, and some ovary genes (RIP2 and PLD1) for senes-
cence that were induced.38,39 Nevertheless, in some genes, these
regulatory changes were reversed by feeding sucrose to the stems,
which partially prevented abortion.38
2.3. Gene expression and regulation of invertase
During plant growth and development, gene expression occurs
in a specic order and is adjusted in response to alterations of
internal and external environmental conditions. In maize, Incw1
is selectively expressed in etiolated shoots, roots, and developing
endosperm, whereas Incw2 is only expressed in shoots and the ba-
sal endosperm transfer cells, but not in roots or the mn1 mutant
endosperm.30 Chourey et al. observed that Incw2 mRNA expressed
maximumly in the basal region (the sugar unloading zone) during
the early phase of cell division and elongation in the endosperm.40
In contrast with Incw2, the highest levels of Incw1 transcripts were
found during the storage phase in both the upper (storage cells)
and lower parts of the endosperm.40 Incw3 expression also has
temporal and spatial characteristics, but Incw4 on the contrary is
constitutively expressed in all vegetative and reproductive tissues
that have been tested.28 Both Ivr1 and Ivr2 mRNAs for the vacuolar
invertase isoforms were abundant in the root tip, very young ker-
nel, silk, anther, and pollen.25
Incw1 expression is regulated by sugars at both the transcrip-
tional and post-transcriptional levels; Sucrose, D-glucose, 3-O-
methylglucose, and 2-deoxyglucose can all induce Incw1-L RNA,
however, only the sucrose and D-glucose can promote the increase
of Incw1-S mRNA, which gives rise to an increase of INCW1 activ-
ity.31 Instead, 3-O-methylglucose and 2-deoxyglucose induce a
greater steady-state level of the Incw1-L mRNA, but this increase
of Incw1-L mRNA does not result in increased INCW1 activity.31
Additionally, the two transcripts, Incw1-S and Incw1-L, are identical
in the coding region and in the positions of their transcription ini-
tiation sites but divergent in the 30 untranslated region.31 These re-
sults suggest that the 30 untranslated region of the Incw1 gene
senses the sugars.31 Similarly, Ivr1 and Ivr2 expressions are also
regulated by sugars. The expression of Ivr1 can be inhibited by su-
gar and up-regulated by the depletion of it, whereas Ivr2 is induced
by increased sugar supply.25 Additionally, studies have shown that
rapid repression of the Ivr1 and Ivr2 genes is induced at the root tip
by low oxygen at both the transcript and enzyme activity levels.41
It has been demonstrated that Ivr2 is specically induced by
water stress.16,42 In situ hybridization studies of water stress
showed that glucose accumulation, invertase protein, and the
Ivr2 transcript were co-localized specically in the bundle sheath
and vascular tissue cells of mature stressed leaves and the primary
root tip.42 However, at the early post-pollination stage (4 days after
pollination), Ivr2 mRNA and the corresponding vacuolar invertase
activity were decreased sharply by water stress in the perianth.43
These results suggest that Ivr2 responds differently to water stress
in different tissues or development stages. As in maize, the activity
of soluble acid invertase in wheat anthers was also shown to de-
crease under water stress and thereafter never recovered.44 In
addition, it was reported that the abscisic acid (ABA) can increase
the transcription level of the Ivr2 gene and vacuolar invertase
activity in excised leaves, but has no effect on the expression of
other invertase genes.16 Similar results were observed in growing
maize plantlets cultured in rooting media with added ABA, where
vacuolar invertase activity and Ivr2 gene expression in the roots
and leaves increased sharply.45
3. Sucrose synthase in maize
3.1. Gene structure and enzymatic properties of sucrose
synthase
Sucrose synthase is a glycosyl transferase that not only converts
sucrose into UDPG and fructose in the presence of UDP, but also
catalyzes sucrose synthesis in a reversible manner, although it is
usually believed that it is primarily involved in sucrose break-
down.19,22 In maize, depending on the ionic species present and
the ionic strength of the solution, sucrose synthase may form tet-
rameric, octameric, and other higher polymeric forms, and all iso-
forms of the enzyme have catalytic activity but show differences in
their specic activity.46 The tetramer is the major form and has the
highest specic activity, so the tetramer could be the native form of
the enzyme.46 There are three genes encoding maize sucrose syn-
thase, Sh1,47 Sus1, and Sus2 (Sus3),48,49 among which the paralo-
gous genes Sh1 and Sus1 encode two biochemically similar
isozymes, SH1 and SUS1 respectively, and Sus2 encodes the SUS2
(SUS3) protein. The Sh1 gene contains 16 exons/14 introns and a
TATA box is located 30 bp upstream of the transcription start site.47
The Sus1 gene consists of 15 exons/14 introns, and its structure is
virtually identical to Sh1only the last intron of Sh1 is missing in
Sus1.50 Alignment of the deduced amino acid sequences of these
genes shows that SH1 has 79% and 70% sequence identity with
SUS1 and SUS2 respectively, and the sequence identity between
 X. Ren, J. Zhang / Carbohydrate Research 368 (2013) 2934
SUS1 and SUS2 is 69%. This suggests that the three sucrose syn-
thase genes have high homology. Several analyses of the SH1 and
SUS1 protein phosphorylation sites show that SH1is phosphory-
lated at Ser10, and SUS1 at Ser15 and Ser170.5153 Research on
the intracellular localization of the sucrose synthase isoforms in
maize has demonstrated that SUS1 and SH1 are associated with
membranes and form SUS1-SH1 homo-oligomers, but SUS1 also
forms a hetero-oligomer with SUS2.53 Subbaiah et al. found that
both SH1 and SUS1 were present in the mitochondria where they
were not involved in sucrose metabolism, while the SUS2 isoform
was not detectable in mitochondria.54 Co-immunoprecipitation
studies indicate that SUS interacts with a voltage-dependent anion
channel in an isoform-specic and anoxia-enhanced manner.54
3.2. Roles of sucrose synthase
Sucrose synthase plays a critical role in starch synthesis because
it can convert sucrose into UDPG and fructose in the presence of
UDP, which is the rst step in the conversion of sucrose to
starch.4,5,7 In transgenic potato plants overexpressing the sucrose
synthase gene, the starch content per plant and the total yield were
signicantly increased compared with control plants.6 In maize,
comparative analyses of the normal and mutant genotypes
Sh1Sus1, sh1Sus1, and sh1sus1 showed that endosperm sucrose syn-
thase activity in the double recessive genotype sh1sus1 reduced by
about 99.5% compared with the Sh1Sus1 genotype and the starch
content in the endosperm of the three genotypes (Sh1Sus1, sh1Sus1,
and sh1sus1) was 100%, 78%, and 53%, respectively.2 The accumula-
tion of SH1 protein always coincides with starch deposition in the
endosperm cells, but in the sh1 mutant endosperm the centrally lo-
cated cells were lost at or during the most critical phase of starch
biosynthesis, and the Sus1-specic cells, including the aleurone
layer and the basal endosperm transfer cells, were not associated
with starch biosynthesis.1 Histochemical assays have shown that
the endosperm sucrose synthase moves gradually from the apical
part of the endosperm to the basal endosperm, and this change is
correlated with the localization of starch synthesis during kernel
development.3
Chourey et al. found that the SH1 isozyme plays a dominant role
in providing the substrate for cellulose biosynthesis.2 In cotton and
wheat, research results also show that sucrose synthase activity is
associated with cellulose biosynthesis.8,9 However, a study by
Alonsoa et al. pointed out that the ux of cell wall synthesis was
increased in maize root tips of a double mutant genotype for su-
crose synthase genes, sh1sus1.55 This result indicates that in maize
root tips SH1 and SUS1 are not specic providers for cellulose bio-
synthesis.55 Additionally, research has found that there is a high
amount of SUS1 protein in maize root cap endopolyploid cells,
which suggests an association between endoreduplication and
the abundance of SUS1 activity.56 There is a lot of evidence demon-
strating that sucrose synthase activity is involved in phloem load-
ing and unloading in many plants.5759 In maize young roots, the
expression of both Sh1 and Sus1 is predominantly in the vascular
cylinder region.1 In transgenic tobacco plants expressing the glucu-
ronidase (GUS) reporter gene driven by the maize Sh1 promoter,
the expression of Sh-GUS was detected only in the phloem cells
but not in any other cell types of the vegetative tissues.71 However,
Hnggi et al. found that there was no appreciable accumulation of
sucrose synthase in the phloem of young maize leaves.60 Thus, the
actual function of sucrose synthase in phloem loading and unload-
ing in maize remains to be claried.
3.3. Gene expression and regulation of sucrose synthase
In maize kernels, the three sucrose synthase genes show differ-
ent expression patterns after pollination, but Sh1 expression is the
31
highest.49 The expression of Sus2 appears to be very constant in
many tissues, including endosperm, embryo, roots, shoots, and in
cell suspensions, with no evidence for it having unique tissue spec-
icity.49 Sus1 mRNA accumulates primarily in the endosperm tis-
sue, excluding the aleurone layer, while Sus2 mRNA accumulates
in the aleurone layer and the embryo.61 During maize kernel ger-
mination, the transcription rate of Sh1 is very high in the etiolated
shoot and the primary root, but if the etiolated seedlings are illu-
minated, the transcript level drops by about 95% in the greening
plant parts.62 In developing maize kernels, at the protein level,
SH1 was found only in the endosperm, while SUS1 was found in al-
most all tissues of the kernel with expression levels specic for cell
type and developmental stage.63
A study by Koch et al. found that the Sh1 mRNA was expressed
under low sugar supply (0.2% glucose), while the Sus1 transcript
level was low or non-detectable under low sugar conditions, and
peaked at greater glucose concentrations (2.0%).64 In intact roots,
sucrose synthase presented in the stele and apex, and while sugar
depletion enhanced sucrose synthase accumulation in peripheral
cells, high sugar levels resulted in its accumulation in all cell
types.64 These results indicate that sugar not only affects related
gene expression but also affects the distribution of sucrose syn-
thase. In Arabidopsis, sucrose and mannitol can increase the tran-
scription of AtSUS1.65 In rice, besides sucrose, ABA can also increase
both sucrose synthase activity and sucrose synthase protein
expression in kernels.66 Additionally, research shows that in rice
sugars can stimulate the accumulation of the RSus1 transcript,
and this accumulation required de novo synthesized proteins.67
Many studies have shown that Sh1 and Sus1 are regulated by
oxygen conditions at the transcriptional and translational levels,
and they display different expression patterns under anoxia and
hypoxia. At the transcriptional level, Sus1 is rapidly up-regulated
by hypoxia, but anoxia has less effect.68 The opposite is true for
Sh1; mRNA abundance is increased strongly under anoxia, but less
so under hypoxia.68 At the protein level, total sucrose synthase
activities rise rapidly under hypoxia but show little signicant
change during anoxia.68 McElfresh and Chourey found that anoxia
induced signicant accumulation of Sh1 mRNA, but there was no
detectable increase in the amount of SUS1 protein.69 Anoxia can
not only affect the expression levels of Sh1 and Sus1, but also in-
duce their cell specic expression. Anoxia induces Sh1 mRNA accu-
mulation in the vascular elements, pith, and epidermis, and the
corresponding SH1 protein is increased in these same tissues and
the root cap, but the increased level of SH1 protein is restricted
to cells within approximately 1 cm of the root apex.70 However,
the SUS1 RNA and protein levels display a slight reduction, except
in the root cap where the SUS1 protein, but not the SUS1 RNA, is
induced.70 In transgenic tobacco plants expressing the GUS repor-
ter gene driven by the maize Sh1 promoter, the Sh1 promoter can
direct cell-type-specic expression of the GUS gene, and GUS
expression was regulated by oxygen conditions in tobacco roots.71
4. Sucrose-phosphate synthase in maize
4.1. Gene structure and enzymatic properties of sucrose-
phosphate synthase
Sucrose-phosphate synthase is primarily responsible for the
synthesis of sucrose. It uses fructose-6-phosphate (F-6-P) and
UDPG as substrates to produce sucrose-phosphate and UDP, and
then the sucrose-phosphate product is degraded by sucrose phos-
phatase (SPP) to produce sucrose; this process is irreversible.18
Maize sucrose-phosphate synthase can be activated by glucose-
6-phosphate (G-6-P), and inhibited by inorganic phosphate (Pi),
but this inhibition occurs only in the presence of G-6-P.72 The en-
zyme is also modulated by light; when harvested from maize
32 X. Ren, J. Zhang / Carbohydrate Research 368 (2013) 2934
leaves in the light it has a higher specic activity than when har-
vested from dark leaves, and the apparent molecular weight of
the light enzyme is higher than that of the dark enzyme.72,73 Light
and dark sucrose-phosphate synthases differ in their afnities for
UDPG in the absence of G-6-P, and both can be activated by G-6-
P; the Km value for UDPG of the light enzyme is lowered by G-6-
P, while the Km for UDPG for the dark enzyme remains un-
changed.72 Huber and Huber pointed out that light/dark signals
modulate sucrose-phosphate synthase activity through protein
phosphorylation, and the major regulatory phosphorylation site
is Ser162 in sucrose-phosphate synthase from maize leaves.18 A
study by Vassey showed that phytochrome also increases the reg-
ulation of sucrose-phosphate synthase activity in maize.74 In addi-
tion, several salts, such as NaCl and KCl, can also improve sucrose-
phosphate synthase activity in maize.75,76 There have been at least
seven sucrose-phosphate synthase identied in the maize genome
to date, named ZmSPS1ZmSPS7, among which ZmSPS1ZmSPS5
have full-length sequences, while ZmSPS6 and ZmSPS7 have only
partial sequences.77 However, at present there are only three maize
sucrose-phosphate synthase genes published in NCBI, and only one
of which is named: sucrose-phosphate synthase1 (Genbank No.:
M97550). The deduced amino acid sequences of the two unnamed
sucrose-phosphate synthase genes (Genbank Nos.: EU967840 and
EU973633) have 62% sequence similarity, but sucrose-phosphate
synthase1 has less than 10% sequence similarity with each of them.
This shows that the sucrose-phosphate synthase1 gene has very
low homology with the other two sucrose-phosphate synthase
genes. At present, there have been many studies on sucrose-phos-
phate synthase1, but none on the other two sucrose-phosphate
synthase genes.
4.2. Roles of sucrose-phosphate synthase
To date, there have been plenty of studies on the biological
function of maize sucrose-phosphate synthase through the transfer
of the maize sucrose-phosphate synthase1 gene cloned by Worrell
et al. to other plants.78 When the maize sucrose-phosphate syn-
thase1 gene driven by a ribulose bisphosphate carboxylase small
subunit promoter was expressed in tomato, total sucrose-phos-
phate synthase activity increased up to six times in the leaves of
transgenic plants compared with wild type plants, and the overex-
pressed sucrose-phosphate synthase caused a reduction of starch
and increase of sucrose in the leaves.78 Another study on trans-
genic tomato expressing the sucrose-phosphate synthase1 gene
from maize also showed that the sucrose-phosphate synthase
activity increased obviously, which resulted in a change of carbon
allocation within the plant, and there was a strong positive corre-
lation between the ratio of sucrose to starch and sucrose-phos-
phate synthase activity in the leaves.79 Research results in
transgenic rice have also proved that sucrose-phosphate synthase
can increase the ratio of sucrose to starch.80 Similarly, in transgenic
potato plants overexpressing the maize sucrose-phosphate syn-
thase1 gene, the sucrose-phosphate synthase activity also signi-
cantly increased in the leaves, which not only increased the ratio
of sucrose to starch but also delayed leaf senescence and increased
the average tuber weight and total yield of the transgenics by at
least 20% compared to the control plants.81 Transgenic tobacco
overexpressing the maize sucrose-phosphate synthase gene1 also
owers earlier and has more owers than the wild type plant.82
In transgenic rice overexpressing the maize sucrose-phosphate
synthase1 gene, the carbon export rate was improved and the
photosynthetic activity increased under elevated [CO2] condi-
tions.83 Additionally, several other studies have shown that su-
crose-phosphate synthase activity is correlated with cellulose
synthesis.8486
4.3. Gene expression and regulation of sucrose-phosphate
synthase
The gene expression proles of maize sucrose-phosphate syn-
thases in different organs or tissues have been investigated, and
the results show that they are expressed in the leaves, ears, and
stems, except for ZmSPS6, which is found almost exclusively in
the ear.77 Lutyya et al. observed that ZmSPS1, 6 and 7 were highly
expressed in the leaves, and ZmSPS2 was expressed highly in pollen
and leaves.77 Gao et al. and Yang et al. found that sucrose-phos-
phate synthase activity shows dynamic variation during maize
growth and development.87,88
The expression of the sucrose-phosphate synthase genes is also
regulated by many chemical substances and environmental sig-
nals.8991 In maize, cold stress can induce ZmSPS6 transcription,
and inhibit ZmSPS2, 4 and 7 transcription, but has no effect on
the expression of ZmSPS1 and ZmSPS3; Drought can induce ZmSPS3
expression, and inhibit ZmSPS1, 4 and 6 expression, but has no ef-
fect on ZmSPS2 and 7 expression; In low nitrogen conditions,
ZmSPS2 expression is inhibited, but ZmSPS3 and 6 are induced,
while the expression of ZmSPS1 and 6 remain unchanged.77 Three
methods were used to test the expression of maize sucrose-phos-
phate synthase genes by Cheng et al.:92 First, immunohistological
analyses showed that in young leaves sucrose-phosphate synthase
protein was primarily present in the bundle-sheath cells, whereas
in mature leaves an equivalent amount of sucrose-phosphate syn-
thase protein was found in both bundle-sheath and mesophyll
cells; Western blotting showed contrary results; no signicant
change in the sucrose-phosphate synthase protein was found in
either young or old leaves in a similar dark treatment; Finally,
northern blotting analyses indicated that sucrose-phosphate syn-
thase gene transcription were obviously reduced after dark treat-
ment.92 Together, their results indicated that the expression of
the maize sucrose-phosphate synthase genes can be regulated at
multiple levels.92
5. Conclusions
Invertase, sucrose synthase, and sucrose-phosphate synthase
are all critical enzymes for sucrose metabolism. Generally speak-
ing, invertase and sucrose synthase are mainly involved in sucrose
degradation, while sucrose-phosphate synthase is responsible for
sucrose synthesis. During maize growth and development, these
enzymes play signicant roles directly or indirectly in organ for-
mation, source/sink activity, the fertility of the kernel, starch and
cellulose synthesis, phloem loading and unloading, carbon alloca-
tion, stress tolerance and so on. The physiological functions of indi-
vidual genes are typically studied by genetic transformation.
However, multiple-gene mutations offer the most direct approach
to study their physiological function. It has been reported that the
three genes Incw2, Sh1 and Sus1 have mutants in maize. However,
so far there has been no report of maize sucrose-phosphate syn-
thase gene mutations. Thus, it remains unknown whether the
physiological functions of the seven ZmSPS genes are equal or
not, and what roles each ZmSPS gene plays in different organs/tis-
sues and developmental stages.
Although the genes for invertase, sucrose synthase, and su-
crose-phosphate synthase have been cloned in maize, some of their
enzymatic properties have been identied preliminarily and the
expression proles of most of the genes are known, the regulatory
mechanisms of the expression of these genes are still poorly under-
stood. To date, only the promoters of Sh1 and Sus1 have been
cloned, and their functions have been studied preliminarily
through linking of the reporter genes b-glucuronidase (GUS) or
neomycin phosphotransferase (NPT) II. In a transient expression
 X. Ren, J. Zhang / Carbohydrate Research 368 (2013) 2934
experiment with the Sh1 promoter, the expression of the reporter
gene NPTII was inhibited shortly after transfection by high extra-
cellular sucrose concentrations, but 35 days after transfection
the NPTII activity raised to 405-fold.93 In an analysis of promoter
deletions, the 20 bp fragment upstream of the Sh1 transcription
start site was sufcient to reproduce the expression prole and
the activity of the full promoter.93 Additionally, the expression of
the GUS reporter gene driven by the Sh1 or Sus1 promoter was
found to be not only tissue-specic but also cell type-specic.71,94
Although, the activities of the Sh1 and Sus1 promoters have been
identied, the cis-acting elements of the Sh1 and Sus1 promoters
have not been studied, and there are no reports about the cis-act-
ing elements of the promoters of the invertase, sucrose-phosphate
synthase, and SUS2 genes in maize.
Sucrose, as the major product of photosynthesis, is also the
main form of long-distance transport and distribution in higher
plants. Sucrose is also an important metabolite and signaling
molecule. Therefore, further clarifying the metabolic network of
sucrose would provide the theoretical support for maize breed-
ing and quality improvement. We believe that the following
points should be the focus of future research emphasis and
trends: rst, further cloning of new genes involved in maize su-
crose metabolism; second, continuing to identify the cis-acting
elements of key gene promoters and clarifying their regulation
mechanisms; third, researching interaction models of key genes;
fourth, identifying and nding transcription factors and regula-
tory elements controlling the transcription and expression of
key genes; fth, identifying some excellent single-nucleotide
polymorphism (SNPs) markers of key genes using association
analysis. Achieving the above research goals would be very use-
ful to facilitate the manipulation of sucrose synthesis and degra-
dation by molecular methods to breed desirable maize
genotypes.
Acknowledgements
This work was supported by the Preferentially Financing
projects of scientic and technological activities of overseas
students in Sichuan province.
References
1. Chen, Y. C.; Chourey, P. S. Theor. Appl. Genet. 1989, 78, 553559.
2. Chourey, P. S.; Taliercio, E. W.; Carlson, S. J.; Ruan, Y. L. Mol. Gen. Genet. 1998,
259, 8896.
3. Wittich, P. E.; Vreugdenhil, D. J. Exp. Bot. 1998, 49, 11631171.
4. Yang, J.; Zhang, J.; Wang, Z.; Xu, G.; Zhu, Q. Plant Physiol. 2004, 135, 16211629.
5. Muoz, F. J.; Baroja-Fernndez, E.; Morn-Zorzano, M. T.; Viale, A. M.;
Etxeberria, E.; Alonso-Casajs, N.; Pozueta-Romero J. Plant Cell Physiol. 2005,
46, 13661376.
6. Baroja-Fernndez, E.; Muoz, F. J.; Montero, M.; Etxeberria, E.; Sesma, M. T.;
Ovecka, M., et al Plant Cell Physiol. 2009, 50, 16511662.
7. Geigenberger, P.; Stitt, M. Planta 1993, 189, 329339.
8. Albrecht, G.; Mustroph, A. Russ J. Plant Physiol. 2003, 50, 813820.
9. Xu, S. M.; Brilla, E.; Llewellyna, D. J.; Furbanka, R. T.; Ruan, Y. L. Mol. Plant 2012,
5, 430441.
10. Huang, D. B.; Tang, C. R. Biotechnol. Bull. 2010, 4, 15 (in chinese).
11. Chiou, T. J.; Bush, D. R. Proc. Natl. Acad. Sci. U.S.A. 1998, 95, 47844788.
12. Takashi, A.; Kouichi, M.; Tatsuhito, F. Plant Cell Physiol. 2005, 46, 937946.
13. Chen, J.; Huang, B. Q.; Li, Y. P.; Du, H.; Gu, Y.; Liu, H. M., et al Carbohydr. Res.
2011, 346, 16841691.
14. Hu, Y. F.; Li, Y. P.; Zhang, J. J.; Liu, H. M.; Chen, Z. Y.; Huang, Y. B. Biotechnol. Lett.
2011, 33, 14651471.
15. Spyropoulos, C. G. J. Exp. Bot. 1982, 33, 12101219.
16. Trouverie, J.; Thvenot, C.; Rocher, J. P.; Sotta, B.; Prioul, J. L. J. Exp. Bot. 2003, 54,
21772186.
17. Gupta, A. K.; Kaur, N. J. Biosci. 2005, 30, 761776.
18. Huber, S. C.; Huber, J. L. Annu. Rev. Plant Biol. 1996, 47, 431445.
19. Sturm, A.; Tang, G. Q. Trends Plant Sci. 1999, 4, 401407.
20. Koch, K. Curr. Opin. Plant Biol. 2004, 7, 235246.
21. Hu, R. F.; Jiang, H.; Li, Y. Y. Northern Horticulture 2012, 01, 167170 (in chinese).
22. Sturm, A. Plant Physiol. 1999, 121, 17.
23. Doehlert, D. C.; Felker, F. C. Physiol. Plant. 1987, 70, 5157.
33
24. Xu, J.; Pemberton, G. H.; Almira, E. C.; McCarty, D. R.; Koch, K. E. Plant Physiol.
1995, 108, 12931294.
25. Xu, J. M.; Avigne, W. T.; McCarty, D.; Koch, K. E. Plant Cell 1996, 8, 12091220.
26. Shanker, S.; Salazar, R. W.; Taliercio, E. W.; Chourey, P. S. Plant Physiol. 1995,
108, 873874.
27. Cheng, W. H.; Taliercio, E. W.; Chourey, P. S. Plant Cell 1996, 8, 971983.
28. Kim, J. Y.; Mahe, A.; Guy, S.; Brangeon, J.; Roche, O.; Chourey, P. S., et al Gene
2000, 245, 89102.
29. Li, L.; Hao, Z. F.; Li, X. H.; Xie, C. X.; Li, M. S.; Zhang, D. G., et al Genetica 2011,
139, 479487.
30. Taliercio, E. W.; Kim, J. Y.; Mah, A.; Shanker, S.; Choi, J.; Cheng, W. H., et al
Plant Physiol. 1999, 155, 197204.
31. Cheng, W. H.; Taliercio, E. W.; Chourey, P. S.; Cheng, W. H. Proc. Natl. Acad. Sci.
U.S.A. 1999, 96, 1051210517.
32. Cheng, W. H.; Chourey, P. S. Theor. Appl. Genet. 1999, 98, 485495.
33. Chourey, P. S. Plant Physiol. 1999, 39, 711.
34. LeClere, S.; Schmelz, E. A.; Chourey, P. S. Phytochemistry 2008, 69, 692699.
35. Chourey, P. S.; Lia, Q. B.; Cevallos-Cevallosc, J. Plant Sci. 2012, 184, 4553.
36. Hanft, J. M.; Jones, R. J. Plant Physiol. 1986, 81, 503510.
37. Andersen, M. N.; Asch, F.; Wu, Y.; Jensen, C. R.; Nsted, H. Plant Physiol. 2002,
130, 591604.
38. Mclaughlin, J.; Boyer, J. S. Ann. Bot. 2004, 94, 675689.
39. Boyer, J. S.; McLaughlin, J. E. J. Exp. Bot. 2007, 58, 267277.
40. Chourey, P. S.; Jain, M.; Li, Q. B.; Carlson, S. J. Planta 2006, 223, 159167.
41. Zeng, Y.; Wu, Y.; Avigne, W. T.; Koch, K. E. Plant Physiol. 1999, 121, 599608.
42. Kim, J. Y.; Mah, A.; Brangeon, J.; Prioul, J. L. Plant Physiol. 2000, 124, 7184.
43. Qin, L.; Trouverie, J.; Chateau-Joubert, S.; Simond-Cte, E.; Thvenot, C.; Prioul,
J. L. Plant Sci. 2004, 166, 371379.
44. Dorion, S.; Lalonde, S.; Saini, H. S. Plant Physiol. 1996, 111, 137145.
45. Trouverie, J.; Chateau-Joubert, S.; Thvenot, C.; Jacquemot, M. P.; Prioul, J. L.
Planta 2004, 219, 894905.
46. Su, J. C.; Preiss, J. Plant Physiol. 1978, 61, 389393.
47. Werr, W.; Frommer, W. B.; Maas, C.; Starlinger, P. EMBO J. 1985, 4, 13731380.
48. Gupta, M.; Chourey, P. S.; Burr, B.; Still, P. E. Plant Mol. Biol. 1988, 10, 215224.
49. Carlson, S. J.; Chourey, P. S.; Helentjaris, T.; Datta, R. Plant Mol. Biol. 2002, 49,
1529.
50. Shaw, J. R.; Ferl, R. J.; Baier, J.; Clairz, D. S.; Carson, C.; McCarty, D. R., et al Plant
Physiol. 1994, 106, 16591665.
51. Huber, S. C.; Huber, J. L.; Liao, P. C.; Gage, D. A.; McMichael, R. J.; Chourey, P. S.,
et al Plant Physiol. 1996, 112, 793802.
52. Hardin, S. C.; Winter, H.; Huber, S. C. Plant Physiol. 2004, 134, 14271438.
53. Duncan, K. A.; Hardin, S. C.; Huber, S. C. Plant Cell Physiol. 2006, 47, 959971.
54. Subbaiah, C. C.; Palaniappan, A.; Duncan, K.; Rhoads, D. M.; Huber, S. C.; Sachs,
M. M. J. Biol. Chem. 2006, 281, 1562515635.
55. Alonsoa, A. P.; Raymonda, P.; Hernoulda, M.; Rondeau-Mourob, C.; Graaf, A.;
Choureyd, P., et al Metab. Eng. 2007, 9, 419432.
56. Kladnik, A.; Vilhar, B.; Chourey, P. S.; Dermastia, M. Can. J. Bot. 2004, 82, 96103.
57. Martin, T.; Frommer, W. B.; Salanoubat, M.; Willmitzer, L. Plant J. 1993, 4, 367377.
58. DAousta, M. A.; Yellea, S.; Nguyen-Quoc, B. Plant Cell 1999, 11, 24072418.
59. Fallahi, H.; Scoeld, G. N.; Badger, M. R.; Chow, W. S.; Furbank, R. T.; Ruan, Y. L.
J. Exp. Bot. 2008, 59, 32833295.
60. Hnggi, E.; Fleming, J. F. Planta 2001, 214, 326329.
61. Rowland, L. J.; Chourey, P. S. Maydica 1990, 35, 373382.
62. Springer, B.; Werr, W.; Starlinger, P.; Bennett, D. C.; Zokolica, M.; Freeling, M.
Mol. Gen. Genet. 1986, 205, 461468.
63. Heinlein, M.; Starlinger, P. Mol. Gen. Genet. 1989, 215, 441446.
64. Koch, K. E.; Nolte, K. D.; Duke, E. R.; McCarty, D. R.; Avigne, W. T. Plant Cell 1992,
4, 5969.
65. Baud, S.; Vaultier, M. N.; Rochat, C. J. Exp. Bot. 2004, 55, 397409.
66. Tang, T.; Xie, H.; Wang, Y.; L, B.; Liang, J. J. Exp. Bot. 2009, 60, 26412652.
67. Liao, Y. C.; Wang, A. Y. Physiol. Plant. 2003, 118, 319327.
68. Zeng, Y.; Wu, Y.; Avigne, W. T.; Koch, K. E. Plant Physiol. 1998, 116, 1573
1583.
69. Mcelfresh, K.; Chourey, P. S. Plant Physiol. 1988, 87, 542546.
70. Rowland, L. J.; Chen, Y. C.; Chourey, P. S. Mol. Gen. Genet. 1989, 218, 3340.
71. Yang, N. S.; Russell, D. Proc. Natl. Acad. Sci. U.S.A. 1990, 87, 41444148.
72. Kalt-Torres, W.; Kerr, P. S.; Huber, S. C. Physiol. Plant. 1987, 70, 653658.
73. Sicher, R. C.; Kremer, D. F. Plant Physiol. 1985, 79, 695698.
74. Vassey, T. L. Plant Physiol. 1988, 88, 540542.
75. Huber, S. C.; Huber, J. L. Plant Cell Physiol. 1991, 32, 327333.
76. Abdel-latif, A. Res. J. Agr. Biol. Sci. 2007, 3, 930933.
77. Lutyya, L. L.; Xu, N. D.; Ordine, R. L.; Morrell, J. A.; Miller, P. W.; Duff, S. M. J.
Plant Physiol. 2007, 164, 923933.
78. Worrell, A. C.; Bruneau, J. M.; Summerfelt, K.; Boersig, M.; Voelker, T. A. Plant
Cell 1991, 3, 11211130.
79. Caltier, N.; Foyer, C. H.; Huber, J.; Voelker, T. A.; Huber, S. C. Plant Physiol. 1993,
101, 535543.
80. Ono, K.; Ishimaru, K.; Aoki, N.; Takahashi, S.; Ozawa, K.; Ohkawa, Y., et al Plant
Prod. Sci. 1999, 2, 172177.
81. Ishimaru, K.; Hirotsu, N.; Kashiwagi, T.; Madoka, Y.; Nagasuga, K.; Ono, K., et al
Plant Prod. Sci. 2008, 11, 104107.
82. Baxter, C. J.; Foyer, C. H.; Turner, J.; Rolfe, S. A.; Quick, W. P. J. Exp. Bot. 2003, 54,
18131820.
83. Babb, V. M.; Haigler, C. H. Plant Physiol. 2001, 127, 12341242.
84. Ono, K.; Sasaki, H.; Hara, T.; Kobayashi, K.; Ishimaru, K. Plant Prod. Sci. 2003, 6,
281286.
34 X. Ren, J. Zhang / Carbohydrate Research 368 (2013) 2934

85. Haigler, C. H.; Singh, B.; Zhang, D.; Hwang, S.; Wu, C.; Cai, X., et al Plant Mol.
Biol. 2007, 63, 815832.
86. Park, J. Y.; Canam, T.; Kang, K. Y.; Ellis, D. D.; Manseld, S. D. Transgenic Res.
2008, 17, 181192.
87. Gao, X. Y.; Xu, F. C.; Zhao, Y. H.; Mei, Z. H. J. South China Agric. Univ. 2001, 22,
4648 (in chinese).
88. Yang, G. D.; Zhao, H. W.; Tan, F. Z.; Liu, X. Y.; Yang, G. B.; He, C. A. Heilongjiang
Agric. Sci. 2006, 5, 1416 (in chinese).
89. Santoiani, C. S.; Tognetti, J. A.; Pontis, H. G.; Salerno, G. L. Physiol. Plant. 1993,
87, 8488.
90. Yang, G. Z.; Zhang, M. F. Northern Horticulture 2006, 5, 4547 (in chinese).
91. Choudhury, S. R.; Roy, S.; Das, R.; Sengupta, D. N. Planta 2008, 229, 207
223.
92. Cheng, W. H.; Im, K. H.; Chourey, P. S. Plant Physiol. 1996, 111, 10211029.
93. Maas, C.; Schaal, S.; Werr, W. EMBO J. 1990, 9, 34473452.
94. Huang, X. F.; NguyenQuoc, B.; Sguin, A.; Yelle, S. Euphytica 1998, 103, 1721.