2013 Ba PDF

JBA-06696; No of Pages 20
Biotechnology Advances xxx (2013) xxxxxx
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Biotechnology Advances
journal homepage: www.elsevier.com/locate/biotechadv
Starch biosynthesis, its regulation and biotechnological approaches to improve

crop yields
Abdellatif Bahaji a,1, Jun Li a,1, ngela Mara Snchez-Lpez a, Edurne Baroja-Fernndez a,
Francisco Jos Muoz a, Miroslav Ovecka a,b, Goizeder Almagro a, Manuel Montero a, Ignacio Ezquer a,
Ed Etxeberria c, Javier Pozueta-Romero a,
a
Instituto de Agrobiotecnologa (CSIC/UPNA/Gobierno de Navarra), Mutiloako etorbidea z/g, 31192 Mutiloabeti, Nafarroa, Spain
b
Centre of the Region Han for Biotechnological and Agricultural Research, Department of Cell Biology, Faculty of Science, Palacky University, lechtitel 11, CZ-783 71 Olomouc, Czech Republic
c
University of Florida, Institute of Food and Agricultural Sciences, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850-2299, USA
a r t i c l e i n f o a b s t r a c t
Available online xxxx Structurally composed of the glucose homopolymers amylose and amylopectin, starch is the main storage carbo-
hydrate in vascular plants, and is synthesized in the plastids of both photosynthetic and non-photosynthetic cells.
Keywords: Its abundance as a naturally occurring organic compound is surpassed only by cellulose, and represents both a
ADPglucose cornerstone for human and animal nutrition and a feedstock for many non-food industrial applications including
CalvinBenson cycle production of adhesives, biodegradable materials, and rst-generation bioethanol. This review provides an up-
Carbohydrate metabolism
date on the different proposed pathways of starch biosynthesis occurring in both autotrophic and heterotrophic
Genetic engineering
Microbial volatiles
organs, and provides emerging information about the networks regulating them and their interactions with the
MIVOISAP environment. Special emphasis is given to recent ndings showing that volatile compounds emitted by microor-
Plantmicrobe interaction ganisms promote both growth and the accumulation of exceptionally high levels of starch in mono- and dicoty-
Endocytosis ledonous plants. We also review how plant biotechnologists have attempted to use basic knowledge on starch
Starch futile cycling metabolism for the rational design of genetic engineering traits aimed at increasing starch in annual crop species.
Sucrose synthase Finally we present some potential biotechnological strategies for enhancing starch content.
2013 Elsevier Inc. All rights reserved.
Contents
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
2. Proposed starch biosynthetic pathways . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
2.1. Starch biosynthesis in leaves . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
2.1.1. A classic model of starch biosynthesis according to which the starch biosynthetic pathway is linked to the CalvinBenson cycle by means of
plastidial phosphoglucose isomerase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
2.1.2. Models of starch biosynthesis according to which the starch biosynthetic pathway is not linked to the CalvinBenson cycle by
means of plastidial phosphoglucose isomerase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
2.2. Starch biosynthesis in heterotrophic organs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
2.2.1. Mechanisms of sucrose unloading in heterotrophic organs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
2.2.2. Proposed mechanisms of sucrosestarch conversion in heterotrophic cells . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
Abbreviations: ADPG, ADPglucose; AGP, ADPG pyrophosphorylase; AGPP, ADPG pyrophosphatase; A/N-inv, alkaline/neutral invertase; BT1, Brittle 1; F6P, fructose-6-phosphate; G1P,
glucose-1-phosphate; G6P, glucose-6-phosphate; GBSS, granule bound starch synthase; GPT, glucose-6-phosphate translocator; GWD, glucan, water dikinase; HvBT1, Hordeum vulgare
BT1; MCF, mitochondrial carrier family; MVs, microbial volatiles; NPP, nucleotide pyrophosphatase/phosphodiesterase; MIVOISAP, MIcrobial VOlatiles Induced Starch Accumulation Pro-
cess; NTRC, NADP-thioredoxin reductase C; OPPP, oxidative pentose phosphate pathway; Pi, orthophosphate; 3PGA, 3-phosphoglycerate; pHK, plastidial hexokinase; pPGI, plastidial
phosphoglucoseisomerase; pPGM, plastidial phosphoglucomutase; PPi, inorganic pyrophosphate; pSP, plastidial starch phosphorylase; RSR1, Rice Starch Regulator1; SuSy, sucrose
synthase; SS, starch synthase; T6P, trehalose-6-phosphate; Trx, thioredoxin; UDPG, UDPglucose; UGP, UDPG pyrophosphorylase; WT, wild type; ZmBT1, Zea mays BT1.
Corresponding author. Tel.: +34 948168009; fax: +34 948232191.
E-mail address: javier.pozueta@unavarra.es (J. Pozueta-Romero).
1
These authors contributed equally to this work.
0734-9750/$ see front matter 2013 Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/j.biotechadv.2013.06.006
Please cite this article as: Bahaji A, et al, Starch biosynthesis, its regulation and biotechnological approaches to improve crop yields, Biotechnol
Adv (2013), http://dx.doi.org/10.1016/j.biotechadv.2013.06.006
2 A. Bahaji et al. / Biotechnology Advances xxx (2013) xxxxxx
3. Microbial volatiles promote both growth and accumulation of exceptionally high levels of starch in mono- and dicotyledonous plants . . . . . . . . 0
4. Biotechnological approaches for increasing starch content in heterotrophic organs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.1. Enhancement of AGP activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.2. Increasing supply of starch precursors to the amyloplast . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.3. Increasing supply of sucrose to heterotrophic cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.4. Enhancement of sucrose breakdown within the heterotrophic cell . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.5. Enhancement of expression of starch synthase class IV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.6. Blocking starch breakdown . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.7. Altering the expression of global regulators . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.8. Enhancing trehalose-6-phosphate content . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.9. Potential strategies for increasing starch content in heterotrophic organs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.9.1. Over-expression of BT1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.9.2. Blocking the activity of ADPG breakdown enzymes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.9.3. Ectopic expression of SuSy in the plastid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
4.9.4. Blocking the synthesis of storage proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
5. Conclusions and main challenges . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
1. Introduction industrial needs and social demands. However, while much is known
about starch metabolism, there are still major gaps in our knowledge
Starch is the main storage carbohydrate in vascular plants. Its abun- that prevent breeders and biotechnologists from advancing in their at-
dance as a naturally occurring compound of living terrestrial biomass is tempts to increase starch content in a predictable way (Chen et al.,
surpassed only by cellulose, and represents the primary source of calo- 2012).
ries in the human diet. Because starch is the principal constituent of In this paper, we rst review recent advances in the biochemistry of
the harvestable organ of many agronomic plants, its synthesis and accu- starch biosynthesis and its regulation, and provide an update on different
mulation also inuences crop yields. proposed pathways of starch biosynthesis occurring in both autotrophic
Synthesized in the plastids of both photosynthetic and non- and heterotrophic organs. Special emphasis is given to recent ndings
photosynthetic cells, starch is an insoluble polyglucan produced by showing that volatile compounds emitted by microorganisms promote
starch synthase (SS) using ADP-glucose (ADPG) as the sugar donor mol- both growth and the accumulation of exceptionally high levels of starch.
ecule. Starch has two major components, amylopectin and amylose, both Finally, we assess the progress in molecular strategies for increasing
of which are polymers of -D-glucose units. Amylose is a linear polymer starch in annual crop species by genetic engineering approaches.
of up to several thousand glucose residues, whereas amylopectin is a
larger polymer regularly branched with -1,6-branch points. These
2. Proposed starch biosynthetic pathways
two molecules are assembled together to form a semi-crystalline starch
granule. The exact proportions of these molecules and the size and
Starch is found in the plastids of photosynthetic and non-
shape of the granule vary between species and between organs of the
photosynthetic tissues. Mature chloroplasts occurring in photosynthet-
same plant. The diversity of both composition and physical parameters
ically active cells possess the capacity of providing energy (ATP) and
of starches from different botanical sources gives rise to their diverse
xed carbon for the synthesis of starch during illumination. By contrast,
processing properties and applications in both non-food sectors such as
production of long-term storage of starch taking place in amyloplasts of
sizing agents in textile and paper industry, adhesive gums, and biode-
reserve organs such as tubers, roots and seed endosperms depends
gradable materials, and in food industries, particularly in bakery, thick-
upon the incoming supply of carbon precursors and energy from the
ening, confectionary and emulsication (Burrell, 2003; Mooney, 2009;
cytosol. This difference between the metabolic capacities of chloroplasts
Slattery et al., 2000). Starch is also used as a feedstock for rst generation
and amyloplasts has led to the generally accepted view that the path-
bioethanol production (Goldemberg, 2007). From an industrial perspec-
way(s) involved in starch production are different in photosynthetic
tive, the utilization of starch as a cheap and renewable polymer and clean
and non-photosynthetic cells.
energy source is becoming increasingly attractive as a consequence of so-
cial concerns about industrial wastes generated from fossil fuels.
In general, the harvested parts of our staple crop plants are hetero- 2.1. Starch biosynthesis in leaves
trophic starch-storing organs. Worldwide annual starch production of
cereal seeds (rice, maize, wheat, barley, etc.) is approximately In leaves, a portion of the photosynthetically xed carbon is retained
2 billion tons, whereas worldwide annual starch production of roots within the chloroplasts during the day to synthesize starch, which is
(e.g. cassava and taro) and tubers (e.g. potato and sweet potato) ex- then remobilized during the subsequent night to support non-
ceeds 700 million tons (Food and Agriculture Organization of the Unit- photosynthetic metabolism and growth by continued export of carbon
ed Nations, http://faostat.fao.org), much of which is destined to non- to the rest of the plant. Due to the diurnal rise and fall cycle of its levels,
food purposes. In 2012 for instance, 75 million tons of starch were foliar starch is termed transitory starch.
extracted and used in many industrial applications (http://www. Transitory starch is a major determinant of plant growth (Sulpice
starch.dk/). The increasing demand of starch from non-food industries, et al., 2009), and its metabolism is regulated to avoid a shortfall of car-
the continuing rapid increase in the world's population, predicted to bon at the end of the dark period. It is typically degraded in a near-linear
reach 9 billion by 2050, and the growing loss of arable land to urbaniza- manner during darkness, with only a small amount remaining at the
tion have spurred on efforts to synthesize plants with higher starch end of the night. When changes in the day length occur that leads to a
content. Therefore, a thorough understanding of the mechanisms in- temporary period of carbon starvation, the carbon budget is rebalanced
volved in starch metabolism and regulation is critically important for by increasing the rate of starch synthesis, decreasing the rate of starch
the rational design of experimental traits aimed at improving yields in breakdown and, consequently, decreasing the rate of growth during
agriculture, and producing more and better polymers that t both darkness. The importance of starch turnover in plant growth and
A. Bahaji et al. / Biotechnology Advances xxx (2013) xxxxxx 3
biomass production is demonstrated by studies of mutants that are de- irreversible upon hydrolytic breakdown of PPi by plastidial alkaline
fective in starch synthesis and mobilization. These mutants grow at the pyrophosphatase.
same rate as wild-type (WT) plants under continuous light or during pPGI is strongly inhibited by light (Heuer et al., 1982) and by
very long days, but show a large inhibition of growth in short days CalvinBenson cycle intermediates such as erythrose-4-P and 3-
(Caspar et al., 1985; Gibon et al., 2004; Lin et al., 1988), which is due phosphoglycerate (3PGA) (the latter being an indicator of photosyn-
to the transient depletion of carbon during the night. thetic carbon assimilation) that accumulate during the day at stro-
Leaf starch mainly accumulates in the photosynthetic palisade and mal concentrations higher than the Ki values for pPGI (Backhausen
spongy mesophyll cells, although it can also be synthesized in epider- et al., 1997; Dietz, 1985; Kelly and Latzko, 1980; Zhou and Cheng,
mal cells, stomatal guard cells and bundle sheath cells surrounding 2008). Although both these characteristics of pPGI, and the low stro-
the vasculature (Tsai et al., 2009). Various mechanisms have been pro- mal G6P/F6P ratio occurring during illumination (Dietz, 1985) would
posed to describe starch synthesis and degradation in leaves. Given that indicate that this enzyme is inactive during illumination (and thus
starch breakdown has been thoroughly discussed in recent reviews not involved in transitory starch biosynthesis), genetic evidence
(Streb and Zeeman, 2012), in the following lines we will focus on de- showing that transitory starch biosynthesis occurs solely by the
scribing, analyzing and comparing the proposed mechanisms of starch pPGI pathway has been obtained from characterization of starch-
synthesis in leaves. decient pPGI mutants from various species (Jones et al., 1986;
Kruckeberg et al., 1989; Kunz et al., 2010; Niewiadomski et al.,
2.1.1. A classic model of starch biosynthesis according to which the starch 2005; Yu et al., 2000).
biosynthetic pathway is linked to the CalvinBenson cycle by means of The classic view of leaf starch biosynthesis illustrated in Fig. 1 also
plastidial phosphoglucose isomerase implies that AGP is the sole source of ADPG, and functions as the
It is widely assumed that the whole starch biosynthetic process re- major regulatory step in the starch biosynthetic process (Neuhaus
sides exclusively in the chloroplast and segregated from the sucrose bio- et al., 2005; Stitt and Zeeman, 2012; Streb and Zeeman, 2012; Streb
synthetic process that takes place in the cytosol (Neuhaus et al., 2005; et al., 2009). This enzyme is a heterotetramer comprising two types of
Stitt and Zeeman, 2012; Streb et al., 2009). According to this classical homologous but distinct subunits, the small (APS) and the large (APL)
view of starch biosynthesis (which is schematically illustrated in subunits (Crevilln et al., 2003, 2005). In Arabidopsis, six genes encode
Fig. 1), starch is considered the end-product of a metabolic pathway proteins with homology to AGP. Two of these genes code for small sub-
that is linked to the CalvinBenson cycle by means of the plastidial units (APS1 and APS2, the latter being in a process of pseudogenization)
phosphoglucose isomerase (pPGI). This enzyme catalyzes the conver- and four (APL1APL4) encode large subunits. Whereas APS1, APL1 and
sion of fructose-6-phosphate (F6P) from the CalvinBenson cycle into APL2 are catalytically active, APL3 and APL4 have lost their catalytic
glucose-6-phosphate (G6P), which is then converted into glucose-1- properties during evolution (Ventriglia et al., 2008). In Arabidopsis, the
phosphate (G1P) by the plastidial phosphoglucomutase (pPGM). large subunits are highly unstable in the absence of small subunits
ADPG pyrophosphorylase (AGP) then converts G1P and ATP into inor- (Wang et al., 1998). Therefore, APS1 T-DNA null mutants contain neither
ganic pyrophosphate (PPi) and ADPG necessary for starch biosynthesis. the large nor the small subunit proteins, which result in a total lack of
These three enzymatic steps are reversible, but the last step is rendered AGP activity (Bahaji et al., 2011a; Ventriglia et al., 2008).
CO2
Triose-P Triose-P
1 1
Calvin-Benson
cycle FBP FBP
2 3 2
F6P
4 F6P
4
G6P
5 G6P
5 Sucrose
G1P
Starch 9 G1P
10 6 8
ADPG 7
UDPG Sucrose-P
Chloroplast Cytosol
Fig. 1. Classic interpretation of the mechanisms of starch and sucrose syntheses in leaves. The enzymes are numbered as follows: 1, 1, fructose-1,6-bisphosphate aldolase; 2, 2, fructose
1,6-bisphosphatase; 3, PPi: fructose-6-P 1-phosphotransferase; 4, 4, PGI; 5, 5, PGM; 6, UGP; 7, sucrose-phosphate-synthase; 8, sucrose phosphate phosphatase; 9, AGP; and 10, SS.
According to this view, the starch biosynthetic process resides exclusively in the chloroplast and is segregated from the sucrose biosynthetic pathway that takes place in the cytosol.
AGP is allosterically activated by 3PGA, and inhibited by Pi APS1 and pPGM antisensed potato plants accumulate low levels of starch
(Kleczkowski, 1999). This property makes the production of ADPG high- when compared with WT leaves (Lytovchenko et al., 2002; Mller-
ly sensitive to changes in photoassimilate (e.g. 3PGA) availability in the Rber et al., 1992; Muoz et al., 2005). Furthermore, Arabidopsis pgm
chloroplast, and helps to coordinate photosynthetic CO2 xation with null mutants (Caspar et al., 1985; Koer et al., 2000), adg1-1 mutants
starch synthesis in leaves. AGP activity is also subjected to APS redox with less than 3% of the WT AGP activity (Lin et al., 1988; Wang
regulation (Hendriks et al., 2003; Kolbe et al., 2005; Li et al., 2012; et al., 1998), and APS1 T-DNA null mutants completely lacking AGP
Michalska et al., 2009). Under oxidizing conditions the two APS subunits activity (Bahaji et al., 2011a; Ventriglia et al., 2008) accumulate
are covalently linked via an intermolecular disulde bridge, thus less than 2% of the WT starch. Further evidence supporting that
forming a stable dimer within the heterotetrameric AGP enzyme (Fu AGP plays a crucial role in starch biosynthesis in leaves comes from
et al., 1998; Hendriks et al., 2003). Conversely, under reductive condi- alkaline pyrophosphatase-silenced plants of Nicotiana benthamiana,
tions APS monomerization is accompanied by activation of AGP activity. since their leaves accumulate ca. 60% of the WT starch (George et al.,
APS dimerization markedly decreases the activity of AGP and alters its 2010).
kinetic properties, making it less sensitive to activation by 3PGA and in-
creasing its Km for ATP (Hendriks et al., 2003). In turn, APS 2.1.2. Models of starch biosynthesis according to which the starch
monomerization activates AGP, which is then sensitive to ne control biosynthetic pathway is not linked to the CalvinBenson cycle by means
through allosteric activation by 3PGA. of plastidial phosphoglucose isomerase
By examining the correlation between APS redox status, AGP activity A vast amount of biochemical and genetic data appear to support the
and starch levels in leaves, different works have provided evidence that starch biosynthetic pathway illustrated in Fig. 1 according to which (a)
transitory starch accumulation is nely regulated by APS post- the whole starch biosynthetic process resides exclusively in the chloro-
translational redox modication in response to environmental inputs plast, (b) pPGI links the CalvinBenson cycle with starch biosynthesis,
such as light, sugars and biotic stress (Gibon et al., 2004; Hendriks and (c) AGP is the sole source of ADPG linked to starch biosynthesis.
et al., 2003; Kolbe et al., 2005; J. Li et al., 2011). However, using aps1 However, in recent years an increasing volume of evidence has been
Arabidopsis plants ectopically expressing a redox-insensitive, mutated provided that supports the occurrence of additional/alternative starch
APS1 form, Li et al. (2012) and Hdrich et al. (2012) have independently biosynthetic pathways involving the cytosolic and plastidial compart-
shown that the rates of transitory starch accumulation in these plants ments wherein the supply of ADPG is not directly linked to the
are comparable to that of WT leaves. In addition, Li et al. (2012) report- CalvinBenson cycle by means of pPGI. Thus, Fettke et al. (2011)
ed WT rates of starch accumulation in aps1 plants expressing in the found that the envelope membranes of chloroplasts in mesophyll cells
chloroplast a redox-insensitive AGP from Escherichia coli, and concluded possess a yet to be identied G1P transport machinery enabling the in-
that post-translational redox modication of APS1 in response to light is corporation of cytosolic G1P into the stroma, which is subsequently
not a major determinant of ne regulation of transitory starch accumu- converted into starch by the stepwise AGP and SS reactions. According
lation in Arabidopsis leaves. to these authors, such mechanism would only allow the accumulation
Plastidial NADP-thioredoxin reductase C (NTRC) plays an im- of 1% of the WT starch, and would explain the occurrence of some
portant role in protecting plants against oxidative stress (Pulido trace amounts of starch in the pPGM mutants (Caspar et al., 1985;
et al., 2010; Serrato et al., 2004). Serrato et al. (2004) and Michalska Muoz et al., 2005; Streb et al., 2009).
et al. (2009) reported that Arabidopsis ntrc mutants grown at Kunz et al. (2010) found that leaves of the pgi1-2 null mutant im-
180 mol photons s 1 m 2 are small and their leaves accumulate paired in pPGI activity accumulate 10% of the WT starch content,
low levels of starch when compared with WT leaves. Furthermore, which is restricted to bundle sheath cells adjacent to the mesophyll
Michalska et al. (2009) reported that ntrc leaves exhibit a decrease in and stomatal guard cells. This mutant exhibits high G6P transport activ-
the extent of APS1 activation (reduction) by light. These authors ity as a consequence of the elevated expression of GPT2 (Kunz et al.,
thus concluded that NTRC is a major determinant of both light- 2010), a gene that codes for a G6P/Pi translocator (GPT) mainly operat-
dependent APS1 redox status and transitory starch accumulation. ing in heterotrophic tissues where the imported G6P can be used for the
However, Li et al. (2012) have recently found that the size of synthesis of starch and fatty acids or to drive the oxidative pentose
Arabidopsis ntrc mutants is similar to that of WT plants when phosphate pathway (Bowsher et al., 2007; Kammerer et al., 1998;
cultured at 90 mol photons s1 m2. Under these conditions, APS1 Kang and Rawsthorne, 1996; L. Zhang et al., 2008). According to Kunz
redox status and starch accumulation rates in leaves of ntrc mutants et al. (2010) the unexpected high levels of starch occurring in leaves
are similar to those of WT leaves, strongly indicating that NTRC plays of pPGI mutants can be ascribed to high GPT2-mediated incorporation
a minor role, if any, in the control of both light-dependent APS1 redox of G6P into the chloroplast of bundle sheath cells adjacent to the meso-
status and transitory starch accumulation in Arabidopsis cultured under phyll and stomatal guard cells, where this hexose-phosphate can be
non-stressing conditions. then converted to starch by means of pPGM, AGP and SS as schematical-
In vitro assays using heterologously expressed potato and pea AGP ly illustrated in Fig. 2. This interpretation, however, is hardly reconcil-
have shown that APS can be activated by plastidial thioredoxins (Trxs) f able with previous studies carried out by the same group showing
and m (Ballicora et al., 2000; Geigenberger et al., 2005), indicating that that the starch-decient phenotype of pPGI mutants can be totally
these proteins could act as major determinants of APS redox status and reverted to WT starch phenotype by the ectopic expression of GPT2
of ne regulation of transitory starch accumulation. However, Li et al. (Niewiadomski et al., 2005), and with the fact that leaves of the pgi1-
(2012) found no differences in the rate of starch accumulation between 2/gpt2 double mutant accumulate as much as 60% of the starch occur-
WT leaves and leaves of homozygous trxf1, trxf2, trxm2 and trxm3 mu- ring in pgi1-2 leaves (Kunz et al., 2010). Furthermore, considering that
tants. Furthermore, Thormhlen et al. (2013) reported that starch levels (a) most of leaf starch accumulates in the mesophyll cells, (b) micro-
in trxf1 leaves were only slightly reduced when compared with WT scopic analyses revealed that stomatal guard cells and bundle sheath
leaves. In addition, Sanz-Barrio et al. (2013) have shown that APS redox cells adjacent to the mesophyll of pPGI mutants accumulate nearly
status in high-starch tobacco leaves overexpressing Trxf and Trxm genes WT starch content (Kunz et al., 2010; Tsai et al., 2009), and (c) guard
from the plastid genome is similar to that of WT leaves. The overall cells of WT leaves possess a G6P transport machinery (Overlach et al.,
data thus indicate that, individually, plastidial Trxs are not major determi- 1993), it is difcult to understand how guard cells and bundle sheath
nants of APS1 redox status-controlled transitory starch content. cells adjacent to the mesophyll of pPGI mutants can accumulate as
Genetic evidence showing that transitory starch biosynthesis occurs much as 10% of the WT starch as a consequence of high GPT2 activity.
solely by the AGP pathway has been obtained from the characterization Needless to say, further investigations will be necessary to understand
of leaves with reduced pPGM and AGP activities. For example, leaves of how and where pgi mutants accumulate ca. 10% of the WT starch.
CO2
Calvin- Triose-P Triose-P

Benson
cycle 1
FBP
3 2
F6P
4
G6P G6P
GPT
5
G1P
Starch 9
10
ADPG
Chloroplast Cytosol
Fig. 2. Suggested model of starch biosynthesis in bundle sheath cells adjacent to the mesophyll and stomatal guard cells of pgi-2 Arabidopsis mutants (from Kunz et al., 2010). Numbering of
enzyme activities is the same as in Fig. 1.
Gerrits et al. (2001) have provided evidence that a sizable pool of analyses (Baroja-Fernndez et al., 2013). Third, HPLC:MS analyses of
sucrose in the plant cell has a plastidial localization. Consistent with the triple ss3/ss4/aps1 mutant impaired in AGP and SS class III and IV re-
the occurrence of a plastidial pool of sucrose, Murayama and Handa vealed that leaves from this mutant accumulate ca. 40-fold more ADPG
(2007) and Vargas et al. (2008) reported the presence of a functional than WT leaves (Ragel, 2012).
alkaline/neutral invertase (A/N-inv) inside plastids. Noteworthy, SuSy catalyzes the reversible conversion of sucrose and NDP into
Vargas et al. (2008) reported that leaves of Arabidopsis mutants impaired fructose and the corresponding NDP-glucose, where N stands for uridine,
in A/N-inv accumulate ca. 70% of the WT starch content. The same au- adenosine, guanosine, cytidine, thymidine or inosine. Although UDP is
thors hypothesized that chloroplastic sucrose plays a sensing role in a generally considered to be the preferred nucleoside diphosphate for
mechanism of control of carbon partitioning and sucrose/starch ratio SuSy, ADP also serves as an effective substrate to produce ADPG
wherein glucose and fructose generated by A/N-inv are sensed through (Baroja-Fernndez et al., 2003, 2012; Cumino et al., 2007; Delmer,
plastidial hexokinase (pHK) (Giese et al., 2005; Olsson et al., 2003). How- 1972; Nakai et al., 1998; Porchia et al., 1999; Silvius and Snyder, 1979;
ever, a possibility cannot be ruled out that, as schematically illustrated in Zervosen et al., 1998). SuSy is highly regulated at both transcriptional
Fig. 3, plastidial sucrose acts as precursor for starch biosynthesis, its and post-transcriptional levels (Asano et al., 2002; Ciereszko and
A/N-inv breakdown products being converted into starch by means Klezkowski, 2002; Djardin et al., 1999; Fu and Park, 1995; Hardin
of pHK, pPGM and AGP. et al., 2004; Koch et al., 1992; Pontis et al., 1981; Purcell et al., 1998; for
Sufcient evidence exists to support the view that a sizable pool of a review see Kleczkowski et al., 2010). Although this sucrolytic enzyme
ADPG in leaves accumulates in the cytosol of photosynthetically compe- is very active in reserve organs, it also expresses in the mesophyll cells
tent cells. This hypothesis is compatible with the occurrence of cytosolic of source leaves (Fu et al., 1995; Wang et al., 1999), its activity in potato
ADPG metabolizing enzymes such as ADPG phosphorylase (McCoy et al., and Arabidopsis leaves greatly exceeding the minimum needed to sup-
2006), glucan synthases (Tacke et al., 1991), sucrose synthase (SuSy) and port normal rate of starch accumulation during illumination (Baroja-
ADPG hydrolases (Olejnik and Kraszewska, 2005; Rodrguez-Lpez et al., Fernndez et al., 2012; Muoz et al., 2005). Accordingly, a metabolic
2000), and with the occurrence in the chloroplast envelope membranes model of transitory starch biosynthesis has been proposed wherein (a)
of yet to be molecularly identied ADPG transport machineries whose both sucrose and starch metabolic pathways are tightly interconnected
activities can account for rates of starch accumulation occurring in leaves by means of cytosolic ADPG producing enzymes such as SuSy (acting
(Pozueta-Romero et al., 1991a). Genetic evidence supporting the occur- when cytosolic sucrose transiently accumulates during illumina-
rence of important ADPG source(s), other than AGP, has been obtained tion), and by the action of an ADPG translocator located at the chlo-
from different sources. First, leaves of transgenic potato and Arabidopsis roplast envelope membranes, and (b) both AGP and pPGM play an
plants ectopically expressing in the cytosol a bacterial ADPG hydrolase important role in the scavenging of glucose units derived from starch
(Moreno-Bruna et al., 2001) accumulate lower ADPG levels than WT breakdown occurring during starch biosynthesis and during the bio-
leaves, as determined by HPLC analyses (Bahaji et al., 2011a; Baroja- genesis of the starch granule (Bahaji et al., 2011a; Baroja-Fernndez
Fernndez et al., 2004). Second, ADPG content in the leaves of AGP and et al., 2005; Muoz et al., 2005, 2006) (Fig. 4). Thus, the net rate of
pPGM mutants is comparable to that of WT leaves, as conrmed by starch accumulation is determined by the balance between the
both HPLC (Bahaji et al., 2011a; Muoz et al., 2005) and HPLC:MS rates of SuSy-mediated ADPG synthesis in the cytosol, import of
CO2
Calvin- Triose-P Triose-P

Benson
cycle 1-8
Sucrose Sucrose
11
Fructose
Glucose
12
12
4
F6P G6P
5
Starch 10 9
ADPG G1P
Chloroplast Cytosol
Fig. 3. Suggested model of starch biosynthesis where sucrose entering the plastid is broken down by A/N-inv to produce fructose and glucose, the latter being converted to G6P by means of
pHK. G6P is subsequently metabolized to starch by the stepwise reactions of pPGM, AGP and SS. Enzyme activities 110 and 4 and 5 are the same as in legend of Fig. 1. Other enzymes
involved are numbered as follows: 11, A/N-inv; 12, pHK.
cytosolic ADPG to the chloroplast, starch synthesis, starch break- warrant starch production and rapid connection of starch metabolism
down and by the efciency with which starch breakdown products with other metabolic pathways.
can be recycled back to starch via the coupled reactions of pPGM The postulated starch biosynthetic pathways involving the plastidial
and AGP. Accordingly, this view predicts that the recovery toward and cytosolic compartments are consistent with still enigmatic results
starch biosynthesis of the glucose units derived from the starch obtained from experiments carried out more than 50 years ago showing
breakdown will be decient in pPGM and AGP mutants, resulting in that 14C in the glucose moiety of sucrose, starch, UDP-glucose and
a parallel decline of starch accumulation. hexose-Ps is asymmetrically distributed in green leaves exposed to
14
The occurrence of a substrate or futile cycle as a result of the simul- CO2 for a short period of time (Gibbs and Kandler, 1957; Havir and
taneous synthesis and breakdown of starch in leaves is not surprising Gibbs, 1963; Kandler and Gibbs, 1956). Unlike the classic view on tran-
since pulse-chase and starch-preloading experiments using isolated sitory starch biosynthesis (Fig. 1) predicting that green leaves exposed to
14
chloroplasts (Fox and Geiger, 1984; Stitt and Heldt, 1981), intact leaves CO2 for a short period of time should synthesize starch with 14C sym-
(Scott and Kruger, 1995; Walters et al., 2004), or cultured photosyn- metrically distributed in the glucose moiety, the proposed mechanisms
thetic cells (Lozovaya et al., 1996) have shown that the illuminated of starch synthesis illustrated in Figs. 24 provide a possible explanation
chloroplasts can synthesize and mobilize starch simultaneously. Fur- for solving the above enigma. As shown in Fig. 4, triose-Ps produced in
thermore, enzymes involved in starch breakdown such as SEX1 and the CalvinBenson cycle are exported to the cytosol where they can be
BAM1 are redox-activated during the light under different environmen- channeled into the oxidative pentose-P pathway (OPPP), thereby leading
tal conditions (Mikkelsen et al., 2005; Sparla et al., 2006; Valerio et al., to a randomization of the carbons giving rise to the asymmetric 14C dis-
2010). Futile cycles lead to a net consumption of ATP, which in some in- tribution observed in hexose-Ps, nucleotide-sugars and sucrose upon ex-
stances can be estimated at about 60% of the total ATP produced by the posure to 14CO2 for a short period of time. Cytosolic G1P, G6P, sucrose
cell (Alonso et al., 2005; Hill and ap Rees, 1994). Although these cycles and/or ADPG will be then transported into the chloroplast and utilized
appear to waste energy without any apparent physiological reason, as precursors for the synthesis of starch molecules possessing glucose
they form part of the organization of the plant central metabolism, molecules with asymmetric 14C distribution.
and contribute to its exibility (Rontein et al., 2002). As presented Genetic evidence consistent with the occurrence of a link between su-
above, starch acts as a major integrator in the plant growth that accu- crose and transitory starch metabolism can be obtained from sucrose-
mulates to cope with temporary starvation imposed by the environ- phosphate-synthase and cytosolic PGM decient plants (Fernie et al.,
ment (Sulpice et al., 2009). It is thus conceivable that highly regulated 2002; Strand et al., 2000), since their leaves contain low levels of both su-
starch futile cycling may entail advantages such as sensitive regulation crose and starch as compared with WT leaves. Further genetic evidence
and rapid metabolic channeling toward various pathways in response can be obtained from sedoheptulose-1,7-bisphosphate overexpressing
to physiological and biochemical needs of the plant. Therefore, due to plants (Lefebvre et al., 2005; Miyagawa et al., 2001), since their leaves
the importance of starch in the overall plant metabolism and growth, are characterized by having high levels of both sucrose and starch when
it is tempting to speculate that both redundancy of ADPG sources and compared with WT leaves. Finally, genetic evidence indicating that leaf
starch futile cycling have been selected during plant evolution to sucrose and starch metabolic pathways are linked by SuSy has been
CO2
Calvin- Triose-P Triose-P OPPP

Benson
cycle
Sulfolipids
Fatty acids, Hexose-Ps
OPPP, etc. 1 - 4 1-8
UDPG
GPT 13 Sucrose
G6P 5 G1P
12 ADP
Glucose 15 16
9 14
Fructose
Starch 10
ADPG ADPG
?
Chloroplast Cytosol
Fig. 4. Suggested interpretation of the mechanism of starch biosynthesis in leaves according to which (a) ADPG is produced in the cytosol and transported to the chloroplast by the action
of a yet to be identied ADPG translocator, (b) AGP and pPGM play an important role in the scavenging of glucose units derived from starch breakdown, and (c) starch metabolism is
connected with other metabolic pathways through the metabolites generated by the starch futile cycle. Enzyme activities 110 and 15 are the same as in legend of Fig. 1. Other enzymes
involved are numbered as follows: 12, pHK; 13, plastidial UGP (Okazaki et al., 2009); 14, plastidial AGPP; 15, pSP (Zeeman et al., 2004); and 16, SuSy. Starch to glucose conversion would
involve the coordinated actions of amylases, isoamylases and disproportionating enzymes (Asatsuma et al., 2005; Fulton et al., 2008; Kaplan and Guy, 2005). Note that starch futile cycling
(indicated with red arrows) may entail advantages such as rapid metabolic channeling toward various pathways (such as fatty acid (Periappuram et al., 2000), OPPP under stress condi-
tions (Zeeman et al., 2004) and sulfolipid biosynthesis (Okazaki et al., 2009)) in response to physiological and biochemical needs. This view predicts that the recovery toward starch bio-
synthesis of the glucose units derived from the starch breakdown will be decient in pPGM and AGP mutants, resulting in a parallel decline of starch accumulation and enhancement of
soluble sugars content since starch breakdown derived products (especially glucose and G6P) will leak out the chloroplast through very active glucose translocator (Cho et al., 2011) and
GPT2 (which is highly expressed in pgi, pgm and agp mutants, Kunz et al., 2010). (For interpretation of the references to color in this gure legend, the reader is referred to the web version
of this article.)
obtained from SuSy-overexpressing potato plants whose leaves accumu- uptake of extracellular sucrose and subsequent transport to the vacuole
late higher ADPG and starch contents than WT leaves (Muoz et al., is not in conict with transport through membrane-bound carriers
2005). Further endeavors based on the characterization of mutants totally given that cell homeostasis can be better maintained if both mechanisms
lacking SuSy will be necessary to conrm (or refute) the involvement of operate in parallel (Etxeberria et al., 2005a). Consistent with this view,
SuSy in starch biosynthesis in leaves. Baroja-Fernndez et al. (2006) reported that carrier-mediated sucrose
import in starved cells of sycamore (Acer pseudoplatanus L.) plays an im-
2.2. Starch biosynthesis in heterotrophic organs portant role in the overall sucrosestarch conversion process during the
initial period of sucrose unloading, whereas endocytosis may play an im-
Sucrose produced in leaves is imported by the heterotrophic organs portant role in transporting the bulk of sucrose to the vacuole and its
and used as carbon source for energy production and starch synthesis in subsequent conversion into starch after prolonged period of sucrose
the amyloplast. As in the case of leaves, various mechanisms have been unloading.
proposed to describe starch biosynthesis in the heterotrophic organs,
which together with the mechanisms of sucrose unloading, will be 2.2.2. Proposed mechanisms of sucrosestarch conversion in heterotrophic
presented and discussed below. cells
Sucrose entering the heterotrophic cell by any of the mechanisms
2.2.1. Mechanisms of sucrose unloading in heterotrophic organs described above must be metabolized to molecules that can be
Unloading of sucrose arriving from aerial parts of the plant into sink transported to the amyloplast for subsequent conversion into starch.
organs occurs either symplastically (moving via plasmodesmata) or Because both chloroplasts and amyloplasts are ontogenically related,
apoplastically. The route depends on the species, organ or tissue. In and because chloroplasts possess a very active triose-P translocator
the apoplastic delivery of sucrose to rapidly growing sinks, apoplastic connecting the plastidial and cytosolic compartments, it was originally
sucrose that has been released into the extracellular space can be split assumed that cytosolic triose-Ps entering the amyloplast by means of
by cell wall-bound invertases into glucose and fructose for subsequent a triose-P translocator act as precursors for starch biosynthesis (Boyer,
uptake into the cell by hexose transporters. Alternatively, sucrose can 1985). However, NMR spectroscopy experiments of starch biogenesis
be taken up by plasma membrane-bound sink specic sucrose trans- in wheat and maize grains using 13C NMR (Hatzfeld and Stitt, 1990;
porters of non-dividing storage sinks (Sauer, 2007). Sucrose is then Keeling et al., 1988) demonstrated that a C-6 molecule, not a triose-P
transported via yet-to-be-identied tonoplast sucrose transporters to (C-3), is the precursor of starch biosynthesis in amyloplasts. Further in-
the vacuole, which serves as temporary reservoir. Apoplastic sucrose vestigations of the enzyme capacities of amyloplasts (Entwistle and ap
can also be taken up by a sucrose-induced endocytic process, and Rees, 1988; Frehner et al., 1990) supported the view that C-6 molecules
transported to the central vacuole of heterotrophic cells (Etxeberria entering amyloplasts are the precursors of starch biosynthesis.
et al., 2005a, 2005b; for a review see Etxeberria et al., 2012). Endocytic Depending on the nature of the C-6 molecule entering the amyloplast
(G6P or ADPG), and depending on the enzymes involved in ADPG regulation (Gmez-Casati and Iglesias, 2002; Hylton and Smith, 1992;
synthesis, various mechanisms have been proposed to explain the Kleczkowski et al., 1993; Weber et al., 1995).
sucrosestarch conversion process in heterotrophic cells. In heterotrophic organs of dicotyledonous plants, sucrose entering the
cytosolic compartment of the heterotrophic cell is broken down by SuSy
2.2.2.1. Models of sucrosestarch conversion in heterotrophic cells to produce fructose and UDPglucose (UDPG), the latter being converted
according to which AGP is the sole source of ADPG linked to starch to G1P and PPi by UDPG pyrophosphorylase (UGP). G1P is subsequently
biosynthesis. It is widely assumed that AGP is the sole source of ADPG metabolized to G6P by means of the cytosolic phosphoglucomutase.
in heterotrophic organs of mono- and dicotyledonous plants. Maximal Cytosolic G6P then enters the amyloplast, where it is converted to starch
in vitro AGP activity greatly exceeds the minimum required to support by the sequential activities of pPGM, AGP and SS (Fig. 5A). Unlike chloro-
the normal rate of starch accumulation in starch storing organs plasts, amyloplasts are unable to photosynthetically generate ATP and
(Denyer et al., 1995; Li et al., 2013; Weber et al., 2000). This would indi- therefore, cytosolic ATP must enter the amyloplast to produce ADPG by
cate that AGP is not a rate-limiting step in the sucrosestarch conver- means of AGP. The presence of an ATP/ADP translocator in the envelope
sion process in many heterotrophic organs. In fact, the ux control membranes of amyloplasts has been rmly established by different labo-
coefcient of AGP has been estimated to be as low as 0.08 in some het- ratories (Pozueta-Romero et al., 1991b; Tjaden et al., 1998), and evidence
erotrophic organs (Denyer et al., 1995; Rolletschek et al., 2002; Weber about its relevance in the starch biosynthetic process has been provided
et al., 2000). As presented above, AGP is a highly regulated enzyme. by Tjaden et al. (1998) and Geigenberger et al. (2001) who reported
However, whereas evidence has been provided that starch syn- that potato tubers with decreased plastidic ATP/ADP transporter activities
thesis is regulated by post-translational redox modication of AGP exhibit reduced starch content.
and 3PGA/Pi balance in potato tubers (Tiessen et al., 2003), AGP in devel- Genetic evidence demonstrating the importance of SuSy in the
oping barley, wheat, pea and bean seeds is insensitive to 3PGA and Pi sucrosestarch conversion process in heterotrophic organs of
10
Starch ADPG
PPi Carriers/Endocytosis
9 Sucrose Sucrose
ATP
UDP
ADP G1P 16
ADP
ATP ATP Fructose
5
5 6
G6P G6P G1P UDPG
GPT
UTP PPi
Amyloplast
Cytosol
Carriers/Endocytosis
Starch Sucrose Sucrose
UDP
16
Fructose
10 9 6
ADPG ADPG G1P UDPG
PPi ATP UTP PPi
Amyloplast
Cytosol
Fig. 5. Classic interpretation of the mechanisms of sucrosestarch conversion in (A) heterotrophic cells of dicotyledonous plants and (B) cereal endosperm cells according to which AGP is
the sole source of ADPG linked to starch biosynthesis. Note that in heterotrophic cells of dicotyledonous plants (a) AGP is exclusively localized in the plastid, (b) the cytosolic carbon sub-
strate compound entering the amyloplast for subsequent conversion into starch is G6P, and (c) cytosolic UGP is involved in the sucroseG6P conversion process. Plastidial and cytosolic
G1P pools are likely connected by means of a yet to be identied G1P translocator occurring in amyloplasts (Fettke et al., 2010). Note also that in cereal endosperms cells (a) most of AGP
has a cytosolic localization, and (b) the cytosolic carbon substrate compound entering the amyloplast for subsequent conversion into starch is ADPG. Numbering of enzyme activities is the
same as in legends of Figs. 14.
dicotyledonous plants comes from the reduced levels of starch in potato sources, other than AGP, of ADPG linked to starch biosynthesis. This hy-
tubers and carrot roots exhibiting low SuSy activity (Tang and Sturm, pothesis is compatible with the occurrence in the amyloplast envelope
1999; Zrenner et al., 1995). In addition, genetic evidence showing that membranes of ADPG transporters (Naeem et al., 1997; Pozueta-
starch biosynthesis in heterotrophic organs involves the incorporation Romero et al., 1991b; Shannon et al., 1998), and with the occurrence
of G6P and its subsequent conversion into ADPG by means of pPGM in heterotrophic organs of cytosolic ADPG metabolizing enzymes such
and AGP has been obtained from the characterization of Vicia as ADPG phosphorylase (Dankert et al., 1964; McCoy et al., 2006;
narbonensis embryos expressing antisense GPT (Rolletschek et al., Murata, 1977) and SuSy. As presented in Section 2.1.2, SuSy is a highly
2007) (these embryos, however, expressed lower levels of starch- regulated enzyme that can produce ADPG from sucrose and ADP. It
related genes such as AGP and SuSy encoding genes), genetically plays a predominant role in determining both sink strength and
engineered pea seeds and potato tubers with reduced pPGM phloem loading, and in the entry of carbon into metabolism in
(Harrison et al., 1998; Tauberger et al., 2000) and from V. narbonensis nonphotosynthetic cells. Maximum in vitro ADPG producing SuSy activ-
seeds and potato tubers with reduced AGP activity (Mller-Rber ity is particularly high in starch storing organs, being comparable to that
et al., 1992; Weber et al., 2000). These organs all exhibit reduced starch of AGP in potato tubers (Baroja-Fernndez et al., 2009) and 34 fold
content when compared with their corresponding WT organs. higher than that of AGP in developing barley and maize endosperms
Using disks from WT and pPGM antisensed potato tubers incubated (Baroja-Fernndez et al., 2003; Li et al., 2013). Furthermore, maximal
with radiolabeled G1P, Fettke et al. (2010) reported that G1P can be ef- ADPG producing SuSy activity in heterotrophic organs such as develop-
ciently taken up by potato tuber parenchyma cells and converted to ing maize endosperm greatly exceeds the minimum required to support
starch. Moreover, these authors reported that exogenously added the normal rate of starch accumulation in this organ (Li et al., 2013),
G1P-dependent starch synthesis was diminished in tuber disks of pota- which would indicate that ADPG producing SuSy activity is not a rate
to plants with low plastidial starch phosphorylase (pSP) activity. Fettke limiting step in the sucrosestarch conversion process in heterotrophic
et al. (2010) thus proposed that G1P can be taken up by amyloplast by a organs.
yet to be identied transporter to be subsequently used as substrate for Essentially in line with investigations describing the occurrence of
pSP- and/or AGP-mediated starch biosynthesis. However, although this simultaneous synthesis and breakdown of glycogen in heterotrophic
hypothesis can explain the results obtained using potato tuber disks, it bacteria and animals (Bollen et al., 1998; David et al., 1990; Gaudet
conicts with previous reports using pSP antisensed potato tubers et al., 1992; Guedon et al., 2000; Massillon et al., 1995; Montero et al.,
showing that, in planta, the lack of pSP did not exert any effect on starch 2011), pulse chase as well as starch pre-loading experiments using iso-
accumulation (Sonnewald et al., 1995). Furthermore, this hypothesis is lated amyloplasts and heterotrophic organs have provided evidence
hardly reconcilable with the strong reduction of starch content in tubers about the occurrence of simultaneous synthesis and breakdown of
of pPGM antisensed potato tubers, indicating that plastidial G6P is starch in non-photosynthetic cells of plants under conditions of active
linked to starch biosynthesis (Tauberger et al., 2000). starch accumulation (Neuhaus et al., 1995; Pozueta-Romero and
Unlike dicotyledonous plants where AGP is exclusively localized in Akazawa, 1993; Sweetlove et al., 1996). The occurrence of starch futile
the plastidial compartment, most of AGP in cereal endosperm cells has cycling in heterotrophic organs has been further fortied by recent
a cytosolic localization (Beckles et al., 2001; Denyer et al., 1996; studies showing that global regulators of storage substance accumula-
Johnson et al., 2003; Kleczkowski, 1996; Thornbjrnsen et al., 1996a, tion such as FLOURY ENDOSPERM2 (FLO2) positively co-regulate the
1996b; Villand and Kleczkowski, 1994). Consistently, most of ADPG has expression of starch synthesis and breakdown genes during starch
a cytosolic localization in cereal endosperm cells (Liu and Shannon, accumulation in developing rice seeds (She et al., 2010) (see
1981; Tiessen et al., 2012). Import studies using amyloplasts isolated Section 4.7), and by the fact that down-regulation of starch breakdown
from maize and barley endosperms of WT plants and starch-decient/ functions in genetically engineered wheat and rice plants results in
ADPG-excess maize Zmbt1 and barley Hvbt1 mutants provided genetic increased grain yield (Hakata et al., 2012; Ral et al., 2012) (see
evidence that Zea mays brittle1 (BT1) (ZmBT1) and Hordeum vulgare Section 4.6). Furthermore, starch breakdown enzymatic activities are
BT1 (HvBT1) are membrane proteins that can incorporate cytosolic high in heterotrophic organs (Li et al., 2013). Consistently, alterna-
ADPG into the amyloplast (Liu et al., 1992; Mhlmann et al., 1997; tive/additional models for the sucrosestarch conversion process in
Patron et al., 2004; Shannon et al., 1996, 1998). Furthermore, cell frac- heterotrophic organs of dicotyledonous and monocotyledonous plants
tionation studies revealed that most of ADPG accumulating in high- have been proposed according to which (a) cytosolic enzymes such as
ADPG developing endosperms of the Riso 13 Hvbt1 mutant has a cyto- SuSy catalyze directly the de novo production from sucrose of ADPG,
solic localization (Tiessen et al., 2012). The overall data thus indicate which is subsequently imported into the amyloplast by the action of
that, in cereal endosperm cells, BT1 proteins facilitate the transfer of an ADPG translocator, and (b) pPGM and AGP play an important role
cytosolic ADPG into the amyloplast for starch biosynthesis and are in the scavenging of the glucose units derived from the starch break-
rate-limiting steps in this process. Based on this evidence, a starch bio- down (Fig. 6). Accordingly, these models preview that the net rate of
synthesis model has been proposed for cereal endosperms wherein the starch accumulation in heterotrophic cells is determined by the balance
stepwise reactions of SuSy, UGP and AGP take place in the cytosol to con- between the rates of ADPG synthesis in the cytosol, import of cytosolic
vert sucrose into ADPG, which enters the amyloplast by means of BT1 to ADPG to the amyloplast, starch synthesis and starch breakdown, and
be utilized as glucosyl donor for starch biosynthesis (Fig. 5B) (Denyer by the efciency with which starch breakdown products can be recycled
et al., 1996; Villand and Kleczkowski, 1994). Genetic evidence demon- back to starch via the coupled reactions of pPGM and AGP. Also, these
strating the importance of SuSy in the sucrosestarch conversion process models predict that the recovery toward starch biosynthesis of starch
in cereal endosperms comes from QTL analyses in maize endosperms breakdown products will be decient in pPGM and AGP mutants,
(Thvenot et al., 2005) and from the reduced levels of starch in maize resulting in a parallel decline of starch accumulation.
seeds exhibiting low SuSy activity (Chourey and Nelson, 1976). Evidence The occurrence of the sucrosestarch conversion pathway illustrat-
showing the importance of AGP in starch biosynthesis in cereal endo- ed in Fig. 6A implies that neither UDPG produced by SuSy, nor cytosolic
sperms comes from AGP mutants exhibiting reduced starch content hexose-Ps derived from the action of UGP on UDPG, are involved in
(Johnson et al., 2003; Tsai and Nelson, 1966). starch biosynthesis in heterotrophic organs of dicotyledonous plants.
This view is consistent with the unexpected WT starch content pheno-
2.2.2.2. Models of sucrosestarch conversion in heterotrophic cells type of UGP antisensed potato tubers (Zrenner et al., 1993), and with
according to which both SuSy and AGP are involved in the synthesis of the WT G6P and G1P contents of starch-decient SuSy antisensed pota-
ADPG linked to starch biosynthesis. Sufcient evidence exists to support to tubers (Baroja-Fernndez et al., 2003). Also, this view is coherent
the view that heterotrophic organs possess in the cytosol important with the unexpected starch-decient phenotype of transgenic potato
Starch 10 16 Carriers/Endocytosis
ADPG ADPG Sucrose Sucrose
Fructose ADP
9
15
Glucose
12
GPT 5
G6P G1P
Amyloplast
Cytosol
Fructose ADP Carriers/Endocytosis

Starch 10
ADPG ADPG Sucrose Sucrose
16
PPi PPi UDP

9 9 16
ATP ATP Fructose
Glucose 15
12
GPT 5 6
G6P G1P G1P UDPG
Amyloplast UTP PPi
Cytosol
Fig. 6. Suggested interpretation of the mechanisms of sucrosestarch conversion in (A) heterotrophic cells of dicotyledonous plants and (B) cereal endosperms according to which (a) both
SuSy and AGP are involved in the synthesis of ADPG linked to starch biosynthesis, (b) the cytosolic carbon substrate compound entering the amyloplast for subsequent conversion into
starch is ADPG, (c) pPGM and AGP are involved in the scavenging of starch breakdown products, and (d) the rate of starch accumulation would be the result of the net balance between
starch synthesis and breakdown. Note that in heterotrophic cells of dicotyledonous plants neither cytosolic hexose-phosphates, nor cytosolic UGP is involved in the sucrosestarch con-
version process. Note also that in cereal endosperm cells SuSy catalyzes the direct conversion of sucrose into both ADPG and UDPG, the latter being converted to ADPG in the cytosol by the
stepwise reactions of UGP and AGP. This view predicts that in pPGM and AGP mutants the recovery toward starch biosynthesis of the glucose units derived from the starch breakdown will
be decient, resulting in a parallel decline of starch accumulation since starch breakdown products (especially glucose and G6P) will leak out the amyloplast. Numbering of enzyme ac-
tivities is the same as in legends of Figs. 14.
tubers heterologously expressing in the cytosol either yeast invertase spp., and from some fungal species inhibit growth in Arabidopsis plants
plus glucokinase (Trethewey et al., 1998) or bacterial sucrose phos- (Splivallo et al., 2007; Tarkka and Piechulla, 2007; Vespermann et al.,
phorylase (Trethewey et al., 2001), since the high sucrolytic activity 2007). Given the lack of knowledge on how microbial volatiles (MVs) af-
occurring in these tubers leads to drastic reduction of cytosolic sucrose fect reprogramming of carbohydrate metabolism in plants, Ezquer et al.
levels, thus limiting ADPG-producing SuSy activity. (2010a) explored the effect on starch metabolism of volatiles released
Genetic evidences demonstrating the signicance of ADPG- from different microbial species ranging from Gram-negative and
producing SuSy in the sucrosestarch conversion process in heterotro- Gram-positive bacteria to different fungi. Toward this end, the authors
phic cells of both mono- and dicotyledonous plants are the same as measured the starch and soluble sugar contents in leaves of plants
those presented in Section 2.2.2.1. cultured in the presence or in the absence of adjacent microbial cultures.
Noteworthy, Ezquer et al. (2010a) reported that, in the absence of phys-
3. Microbial volatiles promote both growth and accumulation of ical contact between plant and microbe, all microbial species tested
exceptionally high levels of starch in mono- and (including plant pathogens and microbes that normally do not interact
dicotyledonous plants with plants) emitted volatiles that promoted enhancement of the intra-
cellular ADPG levels and a rapid accumulation of exceptionally high
Microbes synthesize and emit many volatile compounds. Volatile levels of starch in leaves of both mono- and dicotyledonous plants. Levels
emissions from rhizobacterial isolates of Bacillus spp. promote growth of starch reached by MV-treated leaves were comparable to those occur-
in Arabidopsis plants by facilitating nutrient uptake, photosynthesis and ring in heterotrophic organs such as tubers and cereal seeds. This phe-
defense responses, and by decreasing glucose sensing and abscisic acid nomenon, designated as MIVOISAP (for MIcrobial VOlatiles Induced
levels (Ryu et al., 2003, 2004; H. Zhang et al., 2008, 2009). In contrast, Starch Accumulation Process), was also accompanied by strong promo-
volatiles from Pseudomonas spp., Serratia spp. and Stenotrophomonas tion of growth. Therefore, MIVOISAP cannot be ascribed to utilization
of surplus photosynthates and/or ATP for starch biosynthesis that would unsuccessful, for improving starch accumulation in heterotrophic organs
occur under growth arrest conditions. of plants of agronomic interest. Finally we will present some potential
Transcriptome, metabolite content and enzyme activity analyses of strategies for enhancing starch content.
potato leaves exposed to volatiles emitted by the plant pathogen
Alternaria alternata revealed that starch over-accumulation was accom- 4.1. Enhancement of AGP activity
panied by enhanced 3PGA/Pi ratio and up-regulation of SuSy, acid in-
vertase inhibitors, SSIII and SSIV, starch branching enzyme, GPT2, and Most attempts to increase starch accumulation have focused on en-
synthesis of phosphatidylinositol-phosphates implicated in processes hancing AGP activity. The focus on AGP arose originally from the belief
such as endocytosis and vesicle trafc (Ezquer et al., 2010a). that this enzymatic reaction catalyzes a rate-limiting step in the pro-
MIVOISAP was also accompanied by down-regulation of acid invertase, cess of starch synthesis and thus, increases in its activity would concom-
plastidial Trxs, plastidial -amylase and pSP, and proteins involved in itantly result in enhanced starch accumulation.
the conversion of plastidial triose-phosphates into cytosolic G6P. No One strategy for enhancing AGP activity in plants consisted of heter-
changes were found neither in the expression levels of AGP encoding ologously expressing glgC16 (a mutant variant of the E. coli glgC gene)
genes nor in the redox status of APS1. Furthermore AGP-antisensed po- that encodes an allosterically insensitive AGP form. Recent studies,
tato leaves accumulated exceptionally high levels of starch in the pres- however, have shown that the allosterically sensitive glgC equally
ence of MVs. The overall data thus indicated that potato MIVOISAP is the serves the same purpose, since ectopic glgC expression restores WT
consequence of transcriptionally and post-transcriptionally regulated starch content in aps1 Arabidopsis mutants (Li et al., 2012). First at-
metabolic reprogramming involving (a) up-regulation of ADPG- tempts to increase AGP activity in plants were made in potatoes
producing SuSy, SSIII and SSIV, and proteins involved in the endocytic through expression of glgC16 under the control of the constitutive 35S
uptake and trafc of sucrose, and (b) down-regulation of acid invertase promoter. Using this approach, Stark et al. (1992) reported that when
and starch breakdown enzymes. During potato MIVOISAP there also oc- the product of glgC16 was targeted to plastids, tubers from the trans-
curs a down-regulation of ATP-consuming functions involved in inter- genic lines accumulated more starch than control tubers, although
nal amino acid provision such as proteases and enzymes involved in there was no correlation between starch levels and AGP activity in dif-
de novo synthesis of amino acids. Ezquer et al. (2010a, 2010b) thus hy- ferent lines. Subsequent attempts to reproduce these results on a differ-
pothesized that, under conditions of limited ATP-consuming protein ent cultivar of potato expressing glgC16 under the control of a tuber-
breakdown occurring during exposure to MVs, the surplus ATP and car- specic promoter did not render tubers with increased levels of starch
bon are diverted from protein metabolism to starch biosynthesis. and ADPG, although they showed increased levels of AGP activity
Time-course analyses of starch and maltose contents in illuminated (Sweetlove et al., 1996). Ectopic expression of allosterically insensitive
Arabidopsis leaves of plants exposed to volatiles emitted by A. alternata E. coli AGP has also been used in other crops such as rice (Nagai et al.,
revealed that both starch synthesis and -amylase-dependent starch 2009; Sakulsingharoj et al., 2004), maize (Wang et al., 2007) or cassava
degradation were enhanced upon MV treatment, although the nal (Ihemere et al., 2006). In these cases enhanced AGP activity in seeds
balance was positive for synthesis over breakdown (J. Li et al., 2011). resulted in enhanced biomass, but there was no increase in the starch
The increase of starch content in illuminated leaves of MV-treated hy1/ content on a fresh weight basis.
cry1, hy1/cry2 and hy1/cry1/cry2 Arabidopsis mutants was many-fold Other attempts at achieving increased starch content in cereal endo-
lower than that of WT leaves, indicating that Arabidopsis MIVOISAP is sperms have used modied variants of the AGP large subunit, which
subjected to photoreceptor-mediated control. Transcriptomic analyses contributes little to the catalytic activity of the enzyme but is important
of Arabidopsis leaves exposed to A. alternata volatiles revealed changes in determining its regulatory properties. Giroux et al. (1996) obtained
in the expression of genes involved in multiple processes. However, un- maize lines expressing Sh2r6hs (a mutated variant of the AGP large
like potato, no changes could be observed in the expression of starch re- subunit gene), which encodes an enzyme that is allosterically insensi-
lated genes (J. Li et al., 2011). Also, unlike potato plants, Arabidopsis tive. The starch content of these lines was higher than in control lines;
MIVOISAP was accompanied by 2-3-fold increase of the levels of reduced however there was no increase in starch as a percentage of seed weight
(active) form of APS1. Using different Arabidopsis knockout mutants J. Li and, instead, these lines had signicantly heavier seeds (Giroux et al.,
et al. (2011) observed that the magnitude of the MV-induced starch ac- 1996). Ectopic expression of Sh2r6hs has been conducted in maize,
cumulation was low in mutants impaired in SSIII, SSIV and NTRC. Unlike rice and wheat (Meyer et al., 2004; Smidansky et al., 2002, 2003). In
WT leaves, no changes in the redox status of AGP occurred in ntrc these cases enhanced AGP activity in seeds resulted in increased indi-
mutants, providing evidence that MV-promoted monomerization vidual seed weight and seed yield per plant.
(activation) of AGP is mediated by NTRC. The overall data thus strongly Most recently N. Li et al. (2011) have shown that ectopic expression
indicated that Arabidopsis MIVOISAP involves a photocontrolled, tran- of either the BT2 gene coding for the small subunit maize AGP, or SH2
scriptionally and post-transcriptionally regulated network wherein pho- gene coding for the maize AGP large subunit, results in enhanced seed
toreceptors and post-translational changes (e.g. changes in the redox weight and starch content.
status) of plastidial enzyme(s) such as AGP, SSIII and SSIV play important
roles. Consistent with the possible involvement of changes in redox sta- 4.2. Increasing supply of starch precursors to the amyloplast
tus of SSs in MIVOISAP, Glaring et al. (2012) have recently reported that
SSI and SSIII are reductively activated enzymes. As indicated above, synthesis of storage starch in the amyloplast re-
quires the incorporation of ATP and carbon skeletons from the cytosol.
4. Biotechnological approaches for increasing starch content in Tjaden et al. (1998) and Geigenberger et al. (2001) provided evidence
heterotrophic organs that increasing the supply of ATP to the plastid in potato tubers could
be a strategy to enhance tuber starch content, since transgenic potato
Despite the monumental progress made in understanding the genetic tubers ectopically expressing a plastidic ATP/ADP transporter from
and biochemical mechanisms of starch biosynthesis in plants, up to the Arabidopsis under the control of the cauliower mosaic virus 35S pro-
late 2000s relatively little had been achieved in terms of increasing starch moter accumulated 2-fold more ADPG and 16%36% more starch per
content and yield using biotechnological approaches, particularly in het- gram fresh weight than control tubers. Although these results would
erotrophic organs of cereal crops (Smith, 2008). However, within the last suggest that starch content is very sensitive to changes in the transport
5 years, a series of major developments has produced signicant results machinery that supplies ATP, more recent studies using transgenic po-
that indicate the likelihood of enhancing starch production. We next tato plants ectopically expressing the Arabidopsis ATP/ADP transporter
assess the progress made in molecular strategies, both successful and under the control of a tuber specic promoter did not result in increased
levels of starch (L. Zhang et al., 2008). Furthermore, these studies bacterial sucrose phosphorylase or fructokinase, that facilitate the con-
showed that increasing G6P import to the amyloplast by ectopic expres- version of sucrose or sucrose breakdown products (i.e. glucose and fruc-
sion of a pea GPT had no effect on tuber starch content or tuber yield. By tose) into cytosolic hexose-Ps. However, contrary to expectations,
contrast, double transformants simultaneously expressing the pea GPT transgenic tubers expressing in the cytosol bacterial sucrose phosphor-
and the Arabidopsis ATP/ADP transporter exhibited an enhanced tuber ylase, fructokinase or yeast invertase plus bacterial glucokinase all
yield and starch content, the overall data indicating that both ATP/ADP exhibited reduced starch content (Davies et al., 2005; Trethewey et al.,
and GPT share control of the starch content and tuber yield, most prob- 1998, 2001). Despite extensive research, the mechanisms leading to
ably by co-limiting substrate (ATP and hexose-P) supply to AGP. these unexpected results remain obscure (see Section 2.2.2.2).
Indirect evidence that increasing ATP supply to the amyloplast is a More successful attempts to promote sucrosestarch conversion
strategy for increasing starch content comes from the studies carried have centered on enhancing SuSy activity. The focus on SuSy arose
out by Regierer et al. (2002) who reported that constitutive from the belief that this sucrolytic enzyme is a major determinant of
downregulation of expression of plastidial adenylate kinase, an enzyme sink strength and starch accumulation in both mono- and dicotyledon-
that catalyzes the inter-conversion of ATP with AMP and ADP, increased ous plants. First attempts to increase SuSy activity were conducted in
the ADPG and starch contents of potato tubers (see also patent WO 02/ potatoes through expression of StSUS4 under the control of the constitu-
097101 A1). These changes were accompanied by increases in the total tive 35S promoter. Baroja-Fernndez et al. (2009) reported that UDPG,
adenylate pools (including ATP), although not by changes in the adenyl- ADPG and starch contents in SuSy overexpressing tubers were higher
ate energy charge or by changes in developmental phenotype. Regierer than in WT tubers. Furthermore, starch yield per plant was shown to
et al. (2002) thus hypothesized that adenylate kinase acts in the ATP- be higher in SuSy-overexpressing plants than in WT plants, under
consuming direction, competing for the same ATP pool with AGP. The both greenhouse and open eld conditions (Baroja-Fernndez et al.,
increase in starch content (2 fold) is the largest so far reported for any 2009). High starch SuSy-overexpressing tubers exhibited a trend-
targeted manipulation of starch synthesis. wise increase of AGP activity. Furthermore, they exhibited reduced
acid invertase activity, which together with previous studies showing
4.3. Increasing supply of sucrose to heterotrophic cells that down-regulation of SuSy is accompanied by a concomitant reduc-
tion of starch levels and enhancement of acid invertase activity
The occurrence of high levels of sucrose transporters in heterotrophic (Zrenner et al., 1995) indicates that (a) SuSy, but not acid invertase, is
organs where apoplastic sucrose unloading occurs (e.g. developing rice, involved in the sucrosestarch conversion process, (b) SuSy and acid in-
maize, barley, faba bean, and wheat seeds) has led to the conclusion vertase compete for the same substrate (sucrose), and (c) the balance
that sucrose transporters may play a role in sucrose uptake into endo- between SuSy- and acid invertase-mediated sucrolytic pathways is a
sperm cells for the synthesis of storage compounds during grain lling major determinant of starch accumulation in potato tubers. Interesting-
(Aoki et al., 1999, 2002, 2003; Harrington et al., 1997; Hirose et al., ly enough SuSy overexpressing tubers exhibited other characteristics of
1997; Sivitz et al., 2005; Weschke et al., 2000). Supporting this hypothe- agronomic interest such as substantial increase in antioxidant activity
sis, Scoeld et al. (2002) reported that antisense inhibition of the rice su- and resistance to enzymatic browning when compared to control tu-
crose transporter OsSUT1 leads to impaired grain lling, the nal grain bers. Protection against browning was ascribed to the involvement of
weight being reduced. Toward the aim of evaluating the potential of im- SuSy in the production of UDPG necessary for glycosylation and stabili-
proved sucrose uptake capacity on wheat grain quality and yield zation of polyphenols (patent WO 2010/055186 A1).
Weichert et al. (2010) produced transgenic wheat plants ectopically ex- Most recently Li et al. (2013) produced transgenic maize plants ectop-
pressing the barley sucrose transporter HvSUT1 under the control of an ically expressing StSUS4 under the control of the constitutive UBI promot-
endosperm-specic promoter. The authors reported that, although pro- er. The ve independent transgenic lines characterized produced seeds
tein content in transgenic grains was higher than in WT grains, starch exhibiting higher SuSy activity, higher ADPG content and ca. 10% more
concentration was unchanged. starch than WT seeds at the mature stage. Furthermore, StSUS4 express-
In very early stages of potato tuber development, i.e. during the elon- ing seeds contained a higher amylose/amylopectin balance that WT
gation phase of stolon growth, apoplasmic sucrose unloading predomi- seeds. SuSy (and AGP) activity in WT seeds greatly exceeded the
nates over symplastic unloading. Subsequent tuberization process minimum required to support normal rate of starch accumulation in de-
involves a switch from apoplastic to symplastic phloem unloading veloping maize endosperms (Li et al., 2013). Consequently, SuSy over-
(Viola et al., 2001). Although this would apparently imply that sucrose expression should not lead to enhancement of starch content. To explain
transporters play a minor role in sucrose unloading in developing tubers the observed positive effect of SuSy over-expression on starch content,
and subsequent uptake of sucrose in parenchymatic cells, Khn et al. the authors argued that the rate of starch accumulation is the result of
(2003) reported that reduction of StSUT1 expression in antisensed potato the balance between SuSyAGPADPG transporter-SS-mediated starch
plants leads to reduced yield during early stages of tuber development, synthesis, and amylase- and/or SP-mediated starch breakdown. There-
indicating that in this phase StSUT1 plays an important role for sugar fore, up-regulation of SuSy results in a concomitant increase of the
transport. To evaluate whether it is possible to improve potato plant per- balance between starch synthesis and breakdown, thereby leading to en-
formance by overexpressing a sucrose transporter Leggewie et al. (2003) hancement of starch accumulation. The enhancement of ADPG content in
produced transgenic potato plants heterologously expressing a spinach SuSy over-expressing lines (also occurring in AGP over-expressing rice
sucrose transporter under the control of the 35S promoter. These plants endosperms, Nagai et al., 2009) would indicate that one or more reactions
produced tubers with WT levels of starch content and yield. Thus, the constrain carbon ux from cytosolic ADPG into starch. Li et al. (2013) thus
overall data obtained using both mono- and dicotyledonous crops hypothesized that the transport of ADPG is perhaps the more important
would indicate that up-regulation of sucrose transport is not a satisfacto- limiting factor in starch synthesis in SuSy over-expressing maize
ry strategy for increasing starch content in heterotrophic organs. endosperms.
The overall data thus indicate that enhancement of SuSy activity
4.4. Enhancement of sucrose breakdown within the heterotrophic cell represents a useful strategy for increasing starch accumulation and
yield in heterotrophic organs of both mono- and dicotyledonous crops.
Inspired on the widely accepted view that cytosolic hexose-Ps are
precursors for starch biosynthesis in heterotrophic organs of dicotyle- 4.5. Enhancement of expression of starch synthase class IV
donous plants (see Fig. 5A), attempts to direct more carbon into starch
production in potato tubers consisted of heterologously expressing in Five distinct classes of SSs are known in plants: granule-bound SS
the cytosol enzymes such as yeast invertase plus bacterial glucokinase, (GBSS), which is responsible for the synthesis of amylose, and soluble
SS classes I, II, III, and IV (SSI, SSII, SSIII, and SSIV, respectively), which are dikinases (GWD) involved in starch phosphorylation required for amy-
responsible for the synthesis of amylopectin. Using developing wheat lase mediated starch breakdown are expressed in starch storing organs.
endosperms exposed to SS inactivating temperatures, Keeling et al. Supporting the above hypothesis that reducing starch breakdown
(1993) reported that SSs have a starch biosynthesis ux control coef- would be a viable means of increasing starch content in storage organs
cient approaching to unity. This and the fact that maximum catalytic ac- (see above), Ral et al. (2012) recently reported that RNAi-mediated
tivity of SS is close to metabolic ux through starch biosynthesis in down-regulation of GWD in wheat endosperm under the control of an
several starch storing organs would indicate that SS is a major site of endosperm-specic promoter resulted in an increase in grain yield de-
regulation of starch synthesis, and therefore, a good target for the termined by increases in seed weight, tiller number, spikelets per
production of genetically engineered high-starch plants. However, head and seed number per spike (see also patent WO 2009/067751
ectopic expression of SS genes has hardly ever been used to increase A1). These changes were not accompanied by changes in the percentage
the accumulation of starch. This is mainly due to the presence of differ- of starch per total dried weight, which is a phenomenon comparable to
ent classes of SSs in plants interacting with each other in a protein com- that occurring in AGP overexpressing plants (see above). The work of
plex and displaying a specic role in the synthesis of the nal Ral et al. (2012) provided a promising mechanism for enhancing bio-
architecture of the starch granule (Hennen-Bierwagen et al., 2008). mass and grain yield in wheat, with potential application to other
Consequently, changes in the expression of a single SS may lead to pro- crops. Further evidence supporting that down-regulation of starch deg-
found and unpredictable effects not only on starch structure, but also on radation may contribute to enhance starch content in developing seeds
starch content, revealing that SS isoforms have specic but inter- has been provided by Hakata et al. (2012), who reported that suppres-
dependent roles in starch polymer synthesis. This problem is well sion of -amylase genes improves the quality of rice grain when plants
exemplied by the results obtained using transgenic potato plants ex- are cultured under conditions of high temperature.
pressing the E. coli glycogen synthase (an enzyme that catalyzes the
same reaction as SS) whose tubers accumulated less starch than WT tu- 4.7. Altering the expression of global regulators
bers. Furthermore, the starch from the transgenic tubers had altered
structural properties (Shewmaker et al., 1994). SnRK1 (for Sucrose non-fermenting-1-related kinase-1) is a serine/
Roldn et al. (2007) reported that SSIV T-DNA mutants accumulate threonine protein kinase that forms a complex with sucrose non-
only one large starch granule in Arabidopsis chloroplasts. In addition, fermenting (SNF1) and AMP-activated protein kinase (AMPK). It acts
Szydlowski et al. (2009) reported that Arabidopsis SSIII/SSIV double T- as a central integrator of transcription networks in plant stress and en-
DNA mutants accumulate very reduced starch levels, the overall data ergy signaling, and as such, plays a role in regulating metabolism in re-
indicating that SSIV plays a key role both in the starch granule initiation sponse to carbon availability (Baena-Gonzlez and Sheen, 2008; Baena-
process and in starch accumulation, being mandatory to render the regu- Gonzlez et al., 2007).
lar number of starch granule found in WT plants. That elimination of SSIV Upon sensing the energy decit associated with stress or nutrient
results in both reduction of number of starch granules and starch content deprivation, SnRK1 triggers extensive transcriptional changes that con-
in Arabidopsis leaves suggested that the number of starch granules could tribute to restoring homeostasis. In general, SnRK1 activates genes in-
represent a regulatory step in the control of starch accumulation volved in nutrient remobilization and represses those involved in
(D'Hulst and Mrida, 2010). Conrming this hypothesis Gmez-Arjona biosynthetic processes and storage, thus providing alternative sources
et al. (2011) reported that SSIV overexpression results in enhanced levels of energy through catabolism of amino acids and starch during nutrient
of transitory starch in Arabidopsis leaves. Noteworthy, high levels of starch starvation. Thus, SnRK1 positively regulates the expression of starch
accumulated in the end of the day were completely mobilized during the breakdown genes, since antisense expression of SnRK1 reduces -
night in SSIV overexpressing plants, allowing them to grow at a higher amylase gene expression in cultured wheat and rice embryos during
rate than WT plants. Gmez-Arjona et al. (2011) also reported that ectop- sugar starvation (Laurie et al., 2003; Lu et al., 2007). Intriguingly, al-
ic expression of SSIV results in increased levels of reserve starch and yield though the attributed role of SnRK1 is to repress functions involved in
in tubers of transgenic potato plants cultured both under greenhouse and biosynthetic processes and storage, SnRK1 has been shown to be neces-
open eld conditions. Interestingly enough, SSIV overexpression was sary for starch synthesis, since (a) it is required for SuSy gene expression
accompanied by pleiotropic increase in AGP and SuSy activities, reduction in potato tubers (Debast et al., 2011; Purcell et al., 1998), and (b)
of acid invertase activity and enhancement of ADPG content. Gmez- McKibbin et al. (2006) reported that tubers of SnRK1 overexpressing po-
Arjona et al. (2011) thus hypothesized that enhanced starch accumula- tato plants accumulate up to 30% more starch than control tubers. As
tion in SSIV overexpressing tubers would be the result of enhanced discussed by McKibbin et al. (2006), starch content enhancement
SSIV, AGP and SuSy activities and reduced acid invertase. Further investi- resulting from SnRK1 overexpression can be ascribed to up-regulation
gations will be necessary to determine whether ectopic expression of SSIV of SuSy and AGP activities occurring in the SnRK1 overexpressing
represents a useful strategy for increasing starch content and yield in lines. Although more research using different crop species is necessary
other crops of agronomic interest such as cereal crops. to investigate the role of SnRK1 in the control of starch metabolism,
the work of McKibbin et al. (2006) provided evidence that SnRK1
4.6. Blocking starch breakdown overexpression represents a useful strategy for increasing starch accu-
mulation and yield in heterotrophic organs.
As discussed above (see Section 2.2.2.2), substantial evidence has Genome-wide co-expression analyses conducted to identify candi-
been provided showing the occurrence of simultaneous synthesis and date regulators for starch biosynthesis in rice revealed that Rice Starch
breakdown of starch in some starch accumulating heterotrophic cells. Regulator1 (RSR1), an APETALA2/ethylene-responsive element binding
It is thus highly conceivable that starch accumulated by heterotrophic protein family transcription factor, negatively co-regulates the expres-
organs with active starch turnover is the result of the net balance be- sion of many starch synthesis genes including those coding for AGP,
tween starch synthesis and breakdown (Baroja-Fernndez et al., 2003, SSs, GBSS, branching enzymes and starch debranching enzymes (Fu
2009; Li et al., 2013; Muoz et al., 2006). Consequently, down- and Xue, 2010). RSR1 highly expresses in rice organs and tissues with
regulation of starch breakdown functions should be a viable means of reduced starch content such as roots, seedlings and leaves, and ex-
increasing starch content in heterotrophic organs. presses to much lesser extent in developing seed endosperms. Although
It is generally accepted that -amylase plays a central role in endo- starch content and amylose/amylopectin balance in RSR1 over-
sperm starch degradation. This activity is high in developing seed endo- expressing seeds were shown to be comparable to those of WT seeds,
sperms during the process of starch accumulation (Hakata et al., 2012; seeds of rsr1 mutants displayed increased amylose content as a conse-
Li et al., 2013). On the other hand, genes encoding glucan, water- quence of up-regulation of GBSS expression (Fu and Xue, 2010), had
WT starch content and were larger than WT seeds, a typical phenotype capacities of amyloplasts isolated from Zmbt1 and Hvbt1 seeds indicated
of AGP over-expression (see above). It thus appears that down- that, in cereal endosperms, BT1 proteins facilitate the transfer of
regulation of RSR1 is a good strategy for increasing starch yield in rice. cytosolic ADPG into the amyloplast for starch biosynthesis and are
She et al. (2010) identied FLO2 as a gene highly expressing in devel- rate-limiting steps in this process. This has led to the proposal that up-
oping rice seeds coincident with starch accumulation whose mutation regulation of BT1 function would be a strategy for increasing starch con-
results in both reduced rice grain size and reduced amylose/amylopectin tent in cereal seeds (Chen et al., 2012; Slattery et al., 2000).
balance. The same authors reported that FLO2 positively co-regulates the BT1 proteins belong to the mitochondrial carrier family (MCF) of
expression of genes coding for many starch synthesis enzymes such as proteins that are involved in the transport of metabolites across the mi-
AGP (AGPL14, AGPS1, AGPS2a and AGPS2b), SuSy (SuSy1 and SuSy2), tochondrial membrane (Haferkamp, 2007; Millar and Heazlewood,
SSs (SSIIb, SSIIIb, SSIVa), GBSS and branching enzymes (BE1 and BEIIb), 2003). Although MCF proteins are all presumed to be targeted to the mi-
and enzymes involved in starch breakdown such as -amylase tochondrial inner membrane (Millar and Heazlewood, 2003), some of
(Amy3C, Amy3D and Amy3E), isoamylases (ISA1 and ISA2) and them occur in other subcellular compartments. Kirchberger et al.
pullulanase. Importantly, FLO2 overexpressing rice lines produce larger (2007, 2008) for instance provided evidence that BT1 is exclusively
and heavier grains compared with WT plants, providing evidence that localized to the inner plastidial envelope membranes of plastids. How-
FLO2 overexpression is a good strategy for increasing starch content ever, more recent studies have shown that BT1 proteins are dually
and yield in rice (She et al., 2010). Noteworthy, FLO2 belongs to a family targeted to mitochondria and plastids (Bahaji et al., 2011c).
of rice genes whose orthologs widely exist in plants. Provided the func- Kirchberger et al. (2008) reported that homozygous Atbt1
tion of FLO2 orthologs encoded proteins is similar to that of FLO2, it is Arabidopsis mutants displayed an aberrant growth and sterility pheno-
likely that up-regulation of FLO2 orthologs in other plant species may re- type, which was ascribed to impairments in export of newly synthe-
sult in enhanced starch content and yield. Further research will be neces- sized adenylates from plastids to the cytosol. However, Bahaji et al.
sary to conrm or refute this hypothesis. (2011b) have recently shown that the aberrant growth and sterility
phenotype of Atbt1 mutants could be reverted to WT phenotype by spe-
4.8. Enhancing trehalose-6-phosphate content cic delivery of AtBT1 to mitochondria, indicating that AtBT1 may play
crucial roles, other than transport of newly synthesized adenylates
Trehalose-6-phosphate (T6P) is a sugar signal of emerging signi- from plastids to the cytosol, that are connected to mitochondrial metab-
cance that has been suggested to regulate starch biosynthesis via Trx- olism. Taking into account that (a) similar to AtBT1, ZmBT1 is dually
mediated posttranslational redox activation of AGP in response to sucrose targeted to plastids and mitochondria, (b) similar to homozygous
in leaves, reporting on metabolite status between cytosol and chloroplast Atbt1 Arabidopsis mutants, homozygous Zmbt1 mutant seeds germinate
(Kolbe et al., 2005; Lunn et al., 2006; Schluepmann et al., 2003). poorly and produce plants with low vigor (Mangelsdorf, 1926), (c) mi-
Leaves of transgenic plants with enhanced T6P have increased redox tochondria do not synthesize starch, and (d) there are many low
activation of AGP and increased starch levels, whereas leaves of plants starch maize mutant seeds that germinate and produce plants that
with reduced T6P show the opposite effect (Kolbe et al., 2005). The oc- grow normally, Bahaji et al. (2011a) hypothesized that (a) the aberrant
currence of a positive correlation between intracellular T6P levels, growth and sterility phenotype of homozygous Zmbt1 mutants would
redox AGP activation and rates of starch synthesis in Arabidopsis (Kolbe not be ascribed to starch deciency and/or to reduced ADPG transport
et al., 2005; Lunn et al., 2006), and the promotion of SnRK1-dependent activity in endosperm amyloplasts, and (b) similar to AtBT1, ZmBT1
redox activation of AGP by trehalose or sucrose feeding to potato tuber would be involved in yet to be identied processes occurring in mito-
disks (Tiessen et al., 2003) led to the hypothesis that T6P is a signal of su- chondria, other than ADPG transport, that are important for normal
crose status that promotes starch accumulation in a SnRK1-mediated growth and fertility that indirectly inuence starch accumulation. Need-
process involving redox AGP activation (Geigenberger et al., 2005; less to say further studies are necessary to conrm or refute the latter
Tiessen et al., 2003). However, this proposal conicts with recent nd- hypothesis and to know whether up-regulation of BT1 constitutes a
ings showing that T6P is a strong inhibitor of SnRK1 (Martnez-Barajas valuable strategy for increasing starch content and yields.
et al., 2011; Y. Zhang et al., 2009) and with the fact that genes normally
induced by SnRK1 are repressed by T6P and vice-versa (Y. Zhang et al., 4.9.2. Blocking the activity of ADPG breakdown enzymes
2009). To investigate the role of T6P signaling in starch biosynthesis in An alternative approach to enhancing starch content in crops would
heterotrophic organs Debast et al. (2011) generated transgenic potato be to reduce the rate of ADPG degradation. Plants possess a widely dis-
plants with high levels of T6P as a consequence of the heterologous ex- tributed enzymatic activity, designated as ADPG pyrophosphatase
pression of E. coli OtsA (which encodes a T6P synthase). They also charac- (AGPP), that catalyzes the hydrolytic breakdown of ADPG to G1P and
terized transgenic potato plants with low levels of T6P as a consequence AMP (Rodrguez-Lpez et al., 2000). Two different protein entities are
of the ectopic expression of E. coli OtsB (which encodes a T6P phospha- responsible for this activity: Nudix hydrolases and nucleotide
tase). These authors reported that tubers from transgenic lines with ele- pyrophosphatase/phosphodiesterases (NPP). Nudix hydrolases have
vated T6P levels displayed reduced starch content. On the other hand, been suggested to act as housecleaning enzymes that prevent accu-
tubers from lines with reduced T6P accumulated WT starch content, mulation of reactive nucleoside diphosphate derivates, cell signaling
although they showed a strongly reduced yield as a consequence of molecules or metabolic intermediates by diverting them to metabolic
down-regulation of cell proliferation and growth. Transcriptional prol- pathways in response to biochemical and physiological needs
ing of low T6P tubers revealed that SnRK1 target genes are strongly up- (McLennan, 2006). AGPPs belonging to this family have been found
regulated, which conrms the inhibitory effect of T6P on SnRK1 activity. both in the plastidial and cytosolic compartments (Muoz et al., 2008;
Although Debast et al. (2011) concluded that T6P plays an important role Olejnik and Kraszewska, 2005). Up-regulation of the plastidial AGPP
for potato tuber growth, their results indicated that altering T6P intracel- Nudix hydrolase in transgenic potato plants results in reduced levels
lular levels is not a good strategy for increasing starch content in hetero- of starch content in both leaves and tubers, indicating that this enzyme
trophic organs of plants. has access to the pool of ADPG linked to the biosynthesis of both transi-
tory and reserve starch in leaves and tubers, respectively (Muoz et al.,
4.9. Potential strategies for increasing starch content in heterotrophic organs 2008). The control of the intracellular levels of ADPG linked to starch
production likely represents an important, though still poorly investi-
4.9.1. Over-expression of BT1 gated point of control of starch metabolism. Continued molecular ge-
As pointed out in Section 2.2.2.1, data on ADPG content in seeds of netic analysis will be needed to determine whether AGPPs play a
Zmbt1 maize and Hvbt1 barley mutants, and on ADPG transport crucial role in preventing metabolic ux toward starch in heterotrophic
organs of plants, and whether down-regulation of the AGPP function that autophagy contributes to starch degradation in N. benthamiana
constitutes a good strategy for enhancing starch content and yield in leaves, cytosolic starch granule-like structures being sequestered
major crops. by autophagic bodies and delivered to vacuoles for their subsequent
breakdown. Needless to say, further studies will be necessary to
4.9.3. Ectopic expression of SuSy in the plastid evaluate the relevance of such links in the overall starch metabolism.
As presented in Section 2.1.2., a sizable pool of sucrose has a plastid- Although the study of endocytosis in higher plants remained dor-
ial localization (Gerrits et al., 2001; see Section 2.1.2). SuSys from mant because of the notion that this mechanism would not work
cyanobacteria are highly specic for ADP and have high afnity for against the high turgor pressure in plant cells, mounting evidence
this nucleotide (Curatti et al., 2000; Figueroa et al., 2013; Porchia et al., has been compiled during the last years showing that endocytosis
1999). Thus, a more radical possibility for increasing the starch content is essential for many processes in plants (Mller et al., 2007; Samaj
of crop plants would be to express SuSy (especially from cyanobacterial et al., 2005), including carbohydrate metabolism. We now know
origin) in the plastid to produce ADPG. that in heterotrophic organs where apoplastic sucrose unloading oc-
curs endocytosis may play an important role in transporting the bulk
4.9.4. Blocking the synthesis of storage proteins of sucrose from the apoplast to the vacuole and its subsequent con-
During development of some heterotrophic organs such as tubers version into starch (see Section 2.2.1). Thus, further investigations
and seed endosperms, cell division is followed by cell elongation, differ- will be necessary to understand the molecular and signaling mecha-
entiation and storage. When cell division ceases, a sizable pool of carbon nisms involved in this process.
and ATP is mainly utilized to produce massive amount of storage starch Starch biosynthesis is a highly regulated process, both at transcrip-
and proteins. We must emphasize that a similar phenomenon has been tional and post-transcriptional level that is interconnected with a wide
observed in bacterial cultures entering the stationary phase, where ATP variety of cellular processes and metabolic pathways. Its regulation in-
and carbon are diverted from processes required for cell division and volves a complex and an as yet not well dened assemblage of factors
growth to glycogen production (Montero et al., 2009; Rahimpour that are adjusted to the physiological status of the cell. However, to
et al., 2013). date most studies on genetic regulation of starch metabolism have fo-
Storage protein production is expensive in terms of carbon and ATP cused on the effects of single genes on starch biosynthesis, and analyses
costs and it is highly likely that both storage protein and starch biosyn- of the effect of global regulatory factors on starch metabolism, as well as
thetic processes compete for the same ATP and carbon pools. Therefore, systematic studies of the regulatory mechanisms of starch biosynthesis
blocking storage protein synthesis during the latter phase of develop- are still scanty. Evidence has been provided that, similar to the bacterial
ment could be a potential strategy for increasing starch content in het- system where glycogen synthesis and breakdown genes are co-
erotrophic organs. One way to do it would consist in altering the expressed and co-regulated with genes involved in diverse biological
expression of factors globally controlling the production of storage pro- processes (Montero et al., 2011), starch metabolism genes are co-
teins. This, however, is difcult since it has been reported that the ex- expressed and coordinated with other functions such as amino acid
pression of genes involved in storage starch and protein synthesis is and fatty acid biosynthesis (Fu and Xue, 2010; Giroux et al., 1994; She
highly coordinated by the same factors (Giroux et al., 1994; She et al., et al., 2010; Tsai et al., 2009). Systematic analyses-based characteriza-
2010). One alternative would consist in moderately down-regulating tion of the interactome of starch metabolism-related genes and proteins
the expression of genes directly or indirectly involved in the synthesis with genes and proteins involved in other biological processes, and the
of amino acids by using promoters that are specically expressed during identication of global factors controlling the expression of core starch
the latter part of the heterotrophic organ development. In this respect metabolism functions will be crucially important to rationally design
we must emphasize that accumulation of exceptionally high levels of molecular strategies aimed at enhancing storage starch content in het-
starch in leaves of plants exposed to MVs was accompanied by reduc- erotrophic organs of main crops. In this respect, we must emphasize
tion of the levels of amino acids as a consequence of down-regulation that genome-wide co-expression analyses, which are based on the as-
of genes directly or indirectly involved in amino acid biosynthesis (see sumption that genes with similar expression patterns are more likely
Section 3, Ezquer et al., 2010a). to be functionally related, have proven to be a powerful tool to identify
regulatory factors in transcriptional networks regulating starch metab-
5. Conclusions and main challenges olism in crop species such as rice and potato (Ferreira et al., 2010; Fu
and Xue, 2010). Similar type of comparative transcriptome and prote-
The discrepancies between the different proposed models of ome analyses between different plant organs of WT plants and mutants
starch biosynthesis in leaves (illustrated in Figs. 14) and heterotro- with altered starch content should be conducted using a wide range of
phic organs (Figs. 5 and 6), and the unexpected failures of some mo- species of agronomic interest to identify major determinants and global
lecular strategies aimed at improving storage starch content in regulators of starch biosynthesis.
plants denotes that our knowledge of the starch biosynthetic process MV-promoted accumulation of exceptionally high levels of starch is
is still at a rudimentary level. Better understanding of the regulation a recently discovered mechanism for the elicitation of plant carbohy-
of starch synthesis will require additional research, either at the bio- drate metabolism by microbes that still needs to be investigated in
chemical, cell biological and molecular levels, which in turn will more detail before it can be fully understood and used as a source of
allow plant breeders and biotechnologists to advance in their ideas and tools for strategies to complement breeding and biotechnolo-
attempts to increase starch in a predictable way. Additional knowl- gy programs aimed at improving crop yields. At short run, further efforts
edge on resource allocation within the plant, sucrose uptake in het- and research will be necessary to identify the volatiles and sensing
erotrophic cells, sources of ADPG, mechanisms of ADPG transport mechanisms involved in MIVOISAP.
in plastids, interaction of the plant with both biotic and abiotic envi-
ronmental factors, and metabolic interaction of the plastidial com- Acknowledgments
partment with other cellular compartments will be crucial. In this
last respect we must emphasize that recent studies have demon- This research was partially supported by the grants BIO2010-18239
strated the occurrence of strong, but unexplored metabolic links be- from the Comisin Interministerial de Ciencia y Tecnologa and Fondo
tween plastids and endomembrane organelles. Thus, Nanjo et al. Europeo de Desarrollo Regional (Spain) and IIM010491.RI1 from the
(2006) and Kitajima et al. (2009) have shown that -amylase and Government of Navarra, and by Iden Biotechnology. M.O. was partly
NPPs trafc between endoplasmic reticulum-Golgi and the plastidial supported by grant N. ED0007/01/01 Centre of the Region Han for Bio-
compartment. Furthermore, Wang et al. (2013) have recently shown technological and Agricultural Research.
References Ciereszko I, Klezkowski LA. Glucose and mannose regulate the expression of a major su-
crose synthase gene in Arabidopsis via hexokinase-dependent mechanisms. Plant
Alonso AP, Vigeolas H, Raymond P, Rolin D, Dieuaide-Noubhani M. A new substrate cycle Physiol Biochem 2002;40:90711.
in plants. Evidence for high glucose-phosphate-to-glucose turnover from in vivo Crevilln P, Ballicora MA, Mrida A, Preiss J, Romero JM. The different large subunit
steady-state and pulse-labeling experiments with [13C]glucose and [14C]glucose. isoforms of Arabidopsis thaliana ADP-glucose pyrophosphorylase confer distinct ki-
Plant Physiol 2005;138:222032. netic and regulatory properties to the heterotetrameric enzyme. J Biol Chem
Aoki N, Hirose T, Takahasi S, Ono K, Ishimaru K, Ohsugi R. Molecular cloning and expres- 2003;278:2850815.
sion analysis of a gene for a sucrose transporter in maize (Zea mays L.). Plant Cell Crevilln P, Ventriglia T, Pinto F, Orea A, Mrida A, Romero JM. Differential pattern
Physiol 1999;40:10728. of expression and sugar regulation of Arabidopsis thaliana ADP-glucose
Aoki N, Whitfeld P, Hoeren F, Scoeld G, Newell K, Patrick J, et al. The sucrose transporter pyrophosphorylase-encoding genes. J Biol Chem 2005;280:81439.
genes are expressed in the developing grain of hexaploid wheat. Plant Mol Biol Cumino AC, Marcozzi C, Barreiro R, Salerno GL. Carbon cycling in Anabaena sp. PCC7120. Su-
2002;50:45362. crose synthesis in the heterocysts and possible role in nitrogen xation. Plant Physiol
Aoki N, Hirose T, Scoeld GN, Whitfeld PR, Furbank RT. The sucrose transporter gene family 2007;143:138597.
in rice. Plant Cell Physiol 2003;44:22332. Curatti L, Porchia AC, Herrera-Estrella L, Salerno GL. A prokaryotic sucrose synthase gene
Asano T, Kunieda N, Omura Y, Ibe H, Kawasaki T, Takano M, et al. Rice SPK, a (susA) isolated from a lamentous nitrogen-xing cyanobacterium encodes a protein
calmodulin-like domain protein kinase, is required for storage product accumulation similar to those of plants. Planta 2000;211:72935.
during seed development: phosphorylation of sucrose synthase is a possible factor. D'Hulst C, Mrida A. The priming of storage glucan synthesis from bacteria to plants:
Plant Cell 2002;14:61928. current knowledge and new developments. New Phytol 2010;188:1321.
Asatsuma S, Sawada C, Itoh K, Okito M, Kitajima A, Mitsui T. Involvement of -amylase I-1 Dankert M, Goncalves IRJ, Recondo E. Adenosine diphosphate glucose:orthophosphate
in starch degradation in rice chloroplasts. Plant Cell Physiol 2005;46:85869. adenylyltransferase in wheat germ. Biochim Biophys Acta 1964;81:7885.
Backhausen JE, Jstingmeyer P, Scheibe R. Competitive inhibition of spinach leaf David M, Petit WA, Laughlin MR, Shulman RG, King JE, Barrett EJ. Simultaneous synthesis
phosphoglucose isomerase isoenzymes by erythrose 4-phosphate. Plant Sci 1997;130: and degradation of rat liver glycogen. J Clin Invest 1990;86:6127.
12131. Davies HV, Shepherd LVT, Burrell MM, Carrari F, Urbanczyk-Wochniak E, Leisse A, et al.
Baena-Gonzlez E, Sheen J. Convergent energy and stress signalling. Trends Plant Sci Modulation of fructokinase activity of potato (Solanum tuberosum) results in substan-
2008;13:47482. tial shifts in tuber metabolism. Plant Cell Physiol 2005;46:110315.
Baena-Gonzlez E, Rolland F, Thevelein JM, Sheen J. A central integrator of transcription Debast S, Nunes-Nesi A, Hajirezaei MR, Hofmann J, Sonnewald U, Fernie AR, et al. Altering
networks in plant stress and energy signalling. Nature 2007;448:93843. trehalose-6-phosphate content in transgenic potato tubers affects tuber growth and
Bahaji A, Li J, Ovecka M, Ezquer I, Muoz FJ, Baroja-Fernndez E, et al. Arabidopsis thaliana alters responsiveness to hormones during sprouting. Plant Physiol 2011;156:
mutants lacking ADP-glucose pyrphosphorylase accumulate starch and wild-type 175471.
ADP-glucose content: further evidence for the occurrence of important sources, Djardin A, Sokolov LN, Kleczkowski LA. Sugar/osmoticum levels modulate differential
other than ADP-glucose pyrophosphorylase, of ADP-glucose linked to leaf starch bio- abscisic acid-independent expression of two stress-response sucrose synthase
synthesis. Plant Cell Physiol 2011a;52:116276. genes in Arabidopsis. Biochem J 1999;344:5039.
Bahaji A, Muoz FJ, Ovecka M, Baroja-Fernndez E, Montero M, Li J, et al. Specic delivery Delmer DP. The purication and properties of sucrose synthetase from etiolated Phaseolus
of AtBT1 to mitochondria complements the aberrant growth and sterility phenotype aureus seedlings. J Biol Chem 1972;247:38228.
of homozygous Atbt1 Arabidopsis mutants. Plant J 2011b;68:111521. Denyer K, Foster J, Smith AM. The contributions of ADP-glucose pyrophosphorylase and
Bahaji A, Ovecka M, Brny I, Risueo MC, Muoz FJ, Baroja-Fernndez E, et al. Dual starch-branching enzyme to the control of starch synthesis in developing pea embry-
targeting to mitochondria and plastids of AtBT1 and ZmBT1, two members of the mi- os. Planta 1995;197:5762.
tochondrial carrier family. Plant Cell Physiol 2011c;52:597609. Denyer K, Dunlap F, Thornbjrnsen T, Keeling P, Smith AM. The major form of
Ballicora MA, Frueauf JB, Fu Y, Schrmann P, Press J. Activation of the potato tuber ADP-glucose pyrophosphorylase in maize (Zea mays L.) endosperm is
ADP-glucose pyrophosphorylase by thioredoxin. J Biol Chem 2000;275:131520. extra-plastidial. Plant Physiol 1996;112:77985.
Baroja-Fernndez E, Muoz FJ, Saikusa T, Rodrguez-Lpez M, Akazawa T, Pozueta-Romero Dietz K-J. A possible rate-limiting function of chloroplast hexosemonophosphate isomer-
J. Sucrose synthase catalyzes the de novo production of ADPglucose linked to starch bio- ase in starch synthesis of leaves. Biochim Biophys Acta 1985;839:2408.
synthesis in heterotrophic tissues of plants. Plant Cell Physiol 2003;44:5009. Entwistle G, ap Rees T. Enzymic capacities of amyloplasts from wheat endosperm.
Baroja-Fernndez E, Muoz FJ, Zandueta-Criado A, Morn-Zorzano MT, Viale AM, Biochem J 1988;255:3916.
Alonso-Casajs N, et al. Most of ADPglucose linked to starch biosynthesis occurs out- Etxeberria E, Baroja-Fernndez E, Muoz FJ, Pozueta-Romero J. Sucrose inducible endocy-
side the chloroplast in source leaves. Proc Natl Acad Sci U S A 2004;101:130805. tosis as a mechanism for nutrient uptake in heterotrophic plant cells. Plant Cell Phys-
Baroja-Fernndez E, Muoz FJ, Pozueta-Romero J. A response to Neuhaus et al. Trends iol 2005a;46:47481.
Plant Sci 2005;10:1569. Etxeberria E, Gonzlez P, Pozueta-Romero J. Sucrose transport in Citrus juice cells: evi-
Baroja-Fernndez E, Etxeberria E, Muoz FJ, Morn-Zorzano MT, Alonso-Casajs N, dence for an endocytic transport system. J Am Soc Hortic Sci 2005b;130:26974.
Gonzlez P, et al. An important pool of sucrose linked to starch biosynthesis is Etxeberria E, Pozueta-Romero J, Gonzlez P. In and out of the plant storage vacuole. Plant
taken up by endocytosis in heterotrophic cells. Plant Cell Physiol 2006;47:44756. Sci 2012;190:5261.
Baroja-Fernndez E, Muoz FJ, Montero M, Etxeberria E, Sesma MT, Ovecka M, et al. En- Ezquer I, Li J, Ovecka M, Baroja-Fernndez E, Muoz FJ, Montero M, et al. Microbial volatile
hancing sucrose synthase activity in transgenic potato (Solanum tuberosum L.) tubers emissions promote accumulation of exceptionally high levels of starch in leaves in
results in increased levels of starch, ADPglucose and UDPglucose and total yield. Plant mono- and dicotyledonous plants. Plant Cell Physiol 2010a;51:167493.
Cell Physiol 2009;50:165162. Ezquer I, Li J, Ovecka M, Baroja-Fernndez E, Muoz FJ, Montero M, et al. A suggested
Baroja-Fernndez E, Muoz FJ, Li J, Bahaji A, Almagro G, Montero M, et al. Sucrose synthase model for potato MIVOISAP involving functions of central carbohydrate and amino
activity in the sus1/sus2/sus3/sus4 Arabidopsis mutant is sufcient to support normal acid metabolism, as well as acting cytoskeleton and endocytosis. Plant Signal Behav
cellulose and starch production. Proc Natl Acad Sci U S A 2012;109:3216. 2010b;12:163841.
Baroja-Fernndez E, Bahaji A, Muoz FJ, Li J, Snchez-Lpez AM, Pujol P, et al. HPLC:MS Fernie AR, Tauberger E, Lytovchenko A, Roessner U, Willmitzer L, Trethewey RN. Anti-
analyses of ADPglucose content in mutants lacking plastidial phosphoglucose isomer- sense repression of cytosolic phosphoglucomutase in potato (Solanum tuberosum) re-
ase and ADPglucose pyrophosphorylase conrm the occurrence of various important sults in severe growth retardation, reduction in tuber number and altered carbon
sources of ADPglucose in Arabidopsis. XIII Congresso Luso-Espanhol de Fisiologia Veg- metabolism. Planta 2002;214:51020.
etal. Lisbon, Portugal; 2428 July 2013; 2013. Ferreira SJ, Senning M, Sonnewald S, Kebling P-M, Goldstein R, Sonnewald U. Compara-
Beckles DM, Smith AM, ap Rees T. A cytosolic ADP-glucose pyrophosphorylase is a feature tive transcriptome analysis coupled to X-ray CT reveals sucrose supply and growth
of graminaceous endosperms. Plant Physiol 2001;125:81827. velocity as major determinants of potato tuber starch biosynthesis. BMC Genomics
Bollen M, Keppens S, Stalmans W. Specic features of glycogen metabolism in the liver. 2010;11:93.
Biochem J 1998;336:1931. Fettke J, Albrecht T, Hejazi M, Mahlow S, Nakamura Y, Steup M. Glucose 1-phosphate is
Bowsher CG, Lacey AE, Hanke GT, Clarkson DT, Saker LR, Stulen I, et al. The effect of Glc6P efciently taken up by potato (Solanum tuberosum) tuber parenchyma cells and
uptake and its subsequent oxidation within pea root plastids on nitrite reduction and converted to reserve starch granules. New Phytol 2010;185:66375.
glutamate synthesis. J Exp Bot 2007;58:110918. Fettke J, Malinova I, Albrecht T, Hejazi M, Steup M. Glucose-1-phosphate transport into
Boyer CD. Synthesis and breakdown of starch. In: Neyra CA, editor. Biochemical basis of protoplasts and chloroplasts from leaves of Arabidopsis. Plant Physiol 2011;155:
plant breeding, vol. 1. Boca Raton: CRC Press Inc.; 1985. p. 13353. 172334.
Burrell MM. Starch: the need for improved quality and quantity an overview. J Exp Bot Figueroa CM, Diez MDA, Kuhn ML, McEwen S, Salerno GL, Iglesias AA, et al. The unique
2003;54:4516. nucleotide specicity of the sucrose synthase from Thermosynechococcus elongatus.
Caspar T, Huber SC, Somerville C. Alterations in growth, photosynthesis, and respiration in FEBS Lett 2013;587:1659.
a starchless mutant of Arabidopsis thaliana (L.) decient in chloroplast phosphogluco- Fox TC, Geiger DR. Effects of decreased net carbon exchange on carbohydrate metabolism
mutase activity. Plant Physiol 1985;79:117. in sugar beet source leaves. Plant Physiol 1984;76:7638.
Chen J, Zhang J, Liu H, Hu Y, Huang Y. Molecular strategies in manipulation of the Frehner M, Pozueta-Romero J, Akazawa T. Enzyme sets of glycolysis, gluconeogenesis, and
starch synthesis pathway for improving storage starch content in plants (review oxidative pentose phosphate pathway are not complete in nongreen highly puried
and prospect for increasing storage starch synthesis). Plant Physiol Biochem amyloplasts of sycamore (Acer pseudoplatanus L.) cells suspension cultures. Plant
2012;61:18. Physiol 1990;94:53844.
Cho M-H, Lim H, Shin DH, Jeon J-S, Bhoo SH, Park Y-I, et al. Role of plastidic glucose Fu H, Park WD. Sink- and vascular-associated sucrose synthase functions are encoded by
translocator in the export of starch degradation products from the chloroplasts in different gene classes in potato. Plant Cell 1995;7:136985.
Arabidopsis thaliana. New Phytol 2011;190:10112. Fu F-F, Xue H-W. Coexpression analysis identies rice starch regulator1, a rice AP2/EREBP
Chourey PS, Nelson OE. The enzymatic deciency conditioned by the shrunken-1 muta- family transcription factor, as a novel rice starch biosynthesis regulator. Plant Physiol
tions in maize. Biochem Genet 1976;14:104155. 2010;154:92738.
Fu H, Kim SY, Park WD. A potato Sus3 sucrose synthase gene contains a Hylton C, Smith AM. The rb mutation causes structural and regulatory changes in
context-dependent 3 element and a leader intron with both positive and negative ADPglucose pyrophosphorylase from developing embryos. Plant Physiol 1992;99:
tissue-specic effects. Plant Cell 1995;7:1395403. 162634.
Fu Y, Ballicora MA, Leykam JF, Preiss J. Mechanism of reductive activation of potato tuber Ihemere U, Arias-Garzon D, Lawrence S, Sayre R. Genetic modication of cassava for en-
ADP-glucose pyrophosphorylase. J Biol Chem 1998;273:2504552. hanced starch production. Plant Biotechnol J 2006;4:45365.
Fulton DC, Stettler M, Mettler T, Vaughan CK, Li J, Francisco P, et al. -Amylase4, a Johnson PE, Patron NJ, Bottrill AR, Dinges JR, Fahy BF, Parker ML, et al. A low-starch barley
noncatalytic protein required for starch breakdown, acts upstream of three active mutant, Riso 16, lacking the cytosolic small subunit of ADP-glucose pyrophos-
-amylases in Arabidopsis chloroplasts. Plant Cell 2008;20:104058. phorylase, reveals the importance of the cytosolic isoform and the identity of the
Gmez-Arjona FM, Li J, Raynaud S, Baroja-Fernndez E, Muoz FJ, Ovecka M, et al. En- plastidial small subunit. Plant Physiol 2003;131:68496.
hancing the expression of starch synthase class IV results in increased levels of Jones TWA, Gottlieb LD, Pichersky E. Reduced enzyme activity and starch level in an in-
both transitory and long-term storage starch. Plant Biotechnol J 2011;9:104960. duced mutant of chloroplast phosphoglucose isomerase. Plant Physiol 1986;81:
Gaudet G, Forano E, Dauphin G, Delort A-M. Futile cycling of glycogen in Fibrobacter 36771.
succinogenes as shown by in situ 1H-NMR and 13C-NMR investigation. Eur J Biochem Kammerer B, Fischer K, Hilpert B, Schubert S, Gutensohn M, Weber A, et al. Molecular
1992;207:15562. characterization of a carbon transporter in plastids from heterotrophic tissues: the
Geigenberger P, Stamme C, Tjaden J, Schulz A, Quick PW, Betsche T, et al. Tuber physiol- glucose 6-phosphate/phosphate antiporter. Plant Cell 1998;10:10517.
ogy and properties of starch from tubers of transgenic potato plants with altered Kandler O, Gibbs M. Asymmetric distribution of C14 in the glucose phosphates formed
plastidic adenylate transporter activity. Plant Physiol 2001;125:166778. during photosynthesis. Plant Physiol 1956;31:4112.
Geigenberger P, Kolbe A, Tiessen A. Redox regulation of carbon storage and partitioning in Kang F, Rawsthorne S. Metabolism of glucose-6-phosphate and utilization of multiple me-
response to light and sugars. J Exp Bot 2005;56:146979. tabolites for fatty acid synthesis by plastids from developing oilseed rape embryos.
George GM, van der Merwe MJ, Nunes-Nesi A, Bauer R, Fernie AR, Kossmann J, et al. Planta 1996;199:3217.
Virus-induced gene silencing of plastidial soluble inorganic pyrophosphatase impairs Kaplan F, Guy CL. RNA interference of Arabidopsis beta-amylase8 prevents maltose accu-
essential leaf anabolic pathways and reduces drought stress tolerance in Nicotiana mulation upon cold shock and increases sensitivity of PSII photochemical efciency
benthamiana. Plant Physiol 2010;154:5566. to freezing stress. Plant J 2005;44:73043.
Gerrits N, Turk SCHJ, van Dun KPM, Hulleman SHD, Visser RGF, Weisbeek PJ, et al. Sucrose Keeling PL, Wood JR, Tyson RH, Bridges IG. Starch biosynthesis in developing wheat grain:
metabolism in plastids. Plant Physiol 2001;125:92634. evidence against the direct involvement of triose phosphates in the metabolic path-
Gibbs M, Kandler O. Asymmetric distribution of C14 in sugars formed during photosyn- way. Plant Physiol 1988;87:3119.
thesis. Proc Natl Acad Sci U S A 1957;43:44651. Keeling PL, Bacon PJ, Holt DC. Elevated temperature reduces starch deposition in wheat
Gibon Y, Blsing OE, Palacios-Rojas N, Pankovic D, Hendriks JHM, Fishan J, et al. Adjust- endosperm by reducing the activity of soluble starch synthase. Planta 1993;191:
ment of diurnal starch turnover to short days: depletion of sugar during the night 3428.
leads to a temporary inhibition of carbohydrate utilization, accumulation of sugars Kelly GJ, Latzko E. Oat leaf phosphoglucose isomerase: competitive inhibition by
and post-translational activation of ADP-glucose pyrophosphorylase in the following erythrose-4-phosphate. Photosynth Res 1980;1:1817.
light period. Plant J 2004;39:84762. Kirchberger S, Leroch M, Huynen MA, Wahl M, Neuhaus HE, Tjaden J. Molecular and bio-
Giese J-O, Herbers K, Hoffmann M, Klsgen RB, Sonnewald U. Isolation and functional chemical analysis of the plastidic ADP-glucose transporter (ZmBT1) from Zea mays. J
characterization of a novel plastidic hexokinase from Nicotiana tabacum. FEBS Lett Biol Chem 2007;282:2248191.
2005;579:82731. Kirchberger S, Tjaden J, Neuhaus HE. Characterization of the Arabidopsis Brittle 1 transport
Giroux MJ, Boyer C, Feix G, Hannah LC. Coordinated transcriptional regulation of storage protein and impact of reduced activity on plant metabolism. Plant J 2008;56:5163.
genes in the maize endosperm. Plant Physiol 1994;106:71322. Kitajima A, Asatsuma S, Okada H, Hamada Y, Kaneko K, Nanjo Y, et al. The rice -amylase
Giroux MJ, Shaw J, Barry G, Cobb BG, Greene T, Okita T, et al. A single gene mutation that glycoprotein is targeted from the Golgi apparatus through the secretory pathway to
increases maize seed weight. Proc Natl Acad Sci U S A 1996;93:58249. the plastids. Plant Cell 2009;21:284458.
Glaring MA, Skryhan K, Ktting O, Zeeman SC, Blennow A. Comprehensive survey of Kleczkowski LA. Back to the drawing board: redening starch synthesis in cereals. Trends
redox sensitive starch metabolizing enzymes in Arabidopsis thaliana. Plant Physiol Plant Sci 1996;1:3634.
Biochem 2012;58:8997. Kleczkowski LA. A phosphoglycerate to inorganic phosphate ratio is the major factor in
Goldemberg J. Ethanol for a sustainable energy future. Science 2007;315:80810. controlling starch levels in chloroplasts via ADP-glucose pyrophosphorylase regula-
Gmez-Casati DF, Iglesias AA. ADP-glucose pyrophosphorylase from wheat endosperm. tion. FEBS Lett 1999;448:1536.
Purication and characterization of an enzyme with novel regulatory properties. Kleczkowski LA, Villand P, Lthi E, Olsen O-A, Preiss J. Insensitivity of barley endosperm
Planta 2002;214:42834. ADP-glucose pyrophosphorylase to 3-phosphoglycerate and orthophosphate regula-
Guedon E, Desvaux M, Petitdemange H. Kinetic analysis of Clostridium cellulolyticum car- tion. Plant Physiol 1993;101:17986.
bohydrate metabolism: importance of glucose 1-phosphate and glucose 6-phosphate Kleczkowski LA, Kunz S, Wilczynska M. Mechanisms of UDP-glucose synthesis in plants.
branch points for distribution of carbon uxes inside and outside cells as revealed by Crit Rev Plant Sci 2010;29:191203.
steady-state continuous culture. J Bacteriol 2000;183:20107. Koch KE, Nolte KD, Duke ER, McCarty DR, Avigne WT. Sugar levels modulate differential
Hdrich N, Hendriks JHM, Ktting O, Arrivault S, Feil R, Zeeman SC, et al. Mutagenesis of expression of maize sucrose synthase genes. Plant Cell 1992;4:5969.
cysteine 81 prevents dimerization of the APS1 subunit of ADP-glucose pyrophos- Koer H, Husler RE, Schulz B, Grner F, Flgge U-I, Weber A. Molecular characterization
phorylase and alters diurnal starch turnover in Arabidopsis thaliana leaves. Plant J of a new mutant allele of the plastid phosphoglucomutase in Arabidopsis, and comple-
2012;70:23142. mentation of the mutant with the wild-type cDNA. Mol Gen Genet 2000;262:97886.
Haferkamp I. The diverse members of the mitochondrial carrier family in plants. FEBS Lett Kolbe A, Tiessen A, Schluepmann H, Paul M, Ulrich S, Geigenberger P. Trehalose
2007;581:23759. 6-phosphate regulates starch synthesis via posttranslational redox activation of
Hakata M, Kuroda M, Miyashita T, Yamaguchi T, Kojima M, Sakakibara H, et al. Suppres- ADP-glucose pyrophosphorylase. Proc Natl Acad Sci U S A 2005;102:1111823.
sion of -amylase genes improves quality of rice grain ripened under high tempera- Kruckeberg L, Neuhaus HE, Feil R, Gottlieb LD, Stitt M. Decreased-activity mutants of
ture. Plant Biotechnol J 2012;10:11107. phosphoglucose isomerase in the cytosol and chloroplast of Clarkia xantiana. Biochem
Hardin SC, Winter H, Huber SC. Phosphorylation of the amino terminus of maize sucrose J 1989;261:45767.
synthase in relation to membrane association and enzyme activity. Plant Physiol Khn C, Hajirezaei M-R, Fernie AR, Roessner-Tunali U, Czechowski T, Hirner B, et al. The
2004;134:142738. sucrose transporter StSUT1 localizes to sieve elements in potato tuber phloem and in-
Harrington GN, Nussbaumer Y, Wang X-D, Tegeder M, Franceschi VR, Frommer WB, et al. uences tuber physiology and development. Plant Physiol 2003;131:10213.
Spatial and temporal expression of sucrose transport-related genes in developing Kunz HH, Husler RE, Fettke J, Herbst K, Niewiadomski P, Gierth M, et al. The role of
cotyledons of Vicia faba L. Protoplasma 1997;200:3550. plastidial glucose-6-phosphate/phosphate translocators in vegetative tissues of
Harrison CJ, Hedley CL, Wang TL. Evidence that the rug3 locus of pea (Pisum sativum L.) Arabidopsis thaliana mutants impaired in starch biosynthesis. Plant Biol (Stuttg)
encodes plastidial phosphoglucomutase conrms that the imported substrate for 2010;12(Suppl. 1):11528.
starch synthesis in pea amyloplasts is glucose-6-phosphate. Plant J 1998;13:75362. Laurie S, McKibbin RS, Halford NG. Antisense SNF1-related (SnRK1) protein kinase gene
Hatzfeld W-D, Stitt M. A study of the rate of recycling of triose phosphates in heterotro- represses transient activity of an -amylase (-Amy2) gene promoter in cultured
phic Chenopodium rubrum cells, potato tubers, and maize endosperm. Planta wheat embryos. J Exp Bot 2003;54:73947.
1990;180:198204. Lefebvre S, Lawson T, Fryer M, Zakhleniuk OV, Lloyd JC, Raines CA. Increased
Havir EA, Gibbs M. Studies of the reductive pentose phosphate cycle in intact and sedoheptulose-1,7-bisphosphatase activity in transgenic tobacco plants stimulates
reconstituted chloroplast systems. J Biol Chem 1963;238:31837. photosynthesis and growth from an early stage in development. Plant Physiol
Hendriks JHM, Kolbe A, Gibon Y, Stitt M, Geigenberger P. ADP-glucose pyrophosphorylase 2005;138:45160.
is activated by posttranslational redox-modication in response to light and to sugars Leggewie G, Kolbe A, Lemoine R, Roessner U, Lytovchenko A, Zuther E, et al.
in leaves of Arabidopsis and other plant species. Plant Physiol 2003;133:83849. Overexpression of the sucrose transporter SoSUT1 in potato results in alterations in
Hennen-Bierwagen TA, Liu F, Marsh RS, Kim S, Gan Q, Tetlow IJ, et al. Starch biosynthetic leaf carbon partitioning and in tuber metabolism but has little impact on tuber mor-
enzymes from developing maize endosperm associate in multisubunit complexes. phology. Planta 2003;217:15867.
Plant Physiol 2008;146:1892908. Li J, Ezquer I, Bahaji A, Ovecka M, Baroja-Fernndez E, Muoz FJ, et al. Microbial volatiles
Heuer B, Hansen MJ, Anderson LE. Light modulation of phosphofructokinase in pea leaf induced accumulation of exceptionally high levels of starch in Arabidopsis leaves is a
chloroplasts. Plant Physiol 1982;69:14046. process involving NTRC and starch synthases class III and IV. Mol Plant Microbe Inter-
Hill ST, ap Rees T. Fluxes of carbohydrate metabolism in ripening bananas. Planta act 2011;24:116578.
1994;192:5260. Li N, Zhang S, Zhao Y, Li B, Zhang J. Over-expression of AGPase genes enhances seed
Hirose T, Imaizumi N, Scoeld GN, Furbank RT, Ohsugi R. cDNA cloning and tissue specic weight and starch content in transgenic maize. Planta 2011;233:24150.
expression of a gene for sucrose transporter from rice (Oryza sativa L.). Plant Cell Li J, Almagro G, Muoz FJ, Baroja-Fernndez E, Bahaji A, Montero M, et al. Post-translational
Physiol 1997;38:138996. redox modication of ADP-glucose pyrophosphorylase in response to light is not a
major determinant of ne regulation of transitory starch accumulation in Arabidopsis Muoz FJ, Baroja-Fernndez E, Ovecka M, Li J, Mitsui T, Sesma MT, et al. Plastidial locali-
leaves. Plant Cell Physiol 2012;53:43344. zation of a potato Nudix hydrolase of ADPglucose linked to starch biosynthesis.
Li J, Baroja-Fernndez E, Bahaji A, Muoz FJ, Ovecka M, Montero M, et al. Enhancing su- Plant Cell Physiol 2008;49:173446.
crose synthase activity results in increased levels of starch and ADP-glucose in Murata T. Partial purication and some properties of ADP-glucose phosphorylase from
maize (Zea mays L.) seed endosperms. Plant Cell Physiol 2013;54:28294. potato tubers. Agric Biol Chem 1977;41:19952002.
Lin T-P, Caspar T, Somerville CR, Preiss J. Isolation and characterization of a starchless mu- Murayama S, Handa H. Genes for alkaline/neutral invertase in rice: alkaline/neutral inver-
tant of Arabidopsis thaliana (L.) Heynh lacking ADPglucose pyrophosphorylase activ- tases are located in plant mitochondria and also in plastids. Planta 2007;225:
ity. Plant Physiol 1988;86:11315. 1193203.
Liu T-TY, Shannon JC. Measurement of metabolites associated with nonaqueously isolated Naeem M, Tetlow IJ, Emes MJ. Starch synthesis in amyloplasts puried from developing
starch granules from immature Zea mays L. endosperm. Plant Physiol 1981;67:5259. tubers. Plant J 1997;11:1095103.
Liu K-C, Boyer CD, Shannon JC. Carbohydrate transfer into isolated maize (Zea mays L.) Nagai YS, Sakulsingharoj C, Edwards GE, Satoh H, Greene TW, Blakeslee B, et al. Control of
amyloplasts (abstract no. 234). Plant Physiol 1992;99:S-39. starch synthesis in cereals: metabolite analysis of transgenic rice expressing and
Lozovaya VV, Zabotina OA, Widholm JM. Synthesis and turnover of cell-wall polysaccha- up-regulated cytosplasmic ADP-glucose pyrophosphorylase in developing seeds.
rides and starch in photosynthetic soybean suspension cultures. Plant Physiol Plant Cell Physiol 2009;50:63543.
1996;111:9219. Nakai T, Konishi T, Zhang X-Q, Chollet R, Tonouchi N, Tsuchida T, et al. An increase in ap-
Lu C-A, Lin C-C, Lee K-W, Chen J-L, Huang L-F, Ho S-L, et al. The SnRK1 protein kinase plays parent afnity for sucrose of mung bean sucrose synthase is caused by in vitro phos-
a key role in sugar signaling during germination and seedling growth of rice. Plant phorylation or directed mutagenesis of Ser11. Plant Cell Physiol 1998;39:133741.
Cell 2007;19:248499. Nanjo Y, Oka H, Ikarashi N, Kanedo K, Kitajima A, Mitsui T, et al. Rice plastidial
Lunn J, Feil R, Hendriks JHM, Gibon Y, Morcuende R, Osuna D, et al. Sugar-induced in- N-glycosylated nucleotide pyrophosphatase/phosphodiesterase is transported from
creases in trehalose 6-phosphate are correlated with redox activation of ADPglucose the ER-Golgi to the chloroplast through the secretory pathway. Plant Cell 2006;18:
pyrophosphorylase and higher rates of starch synthesis in Arabidopsis thaliana. 258292.
Biochem J 2006;397:13948. Neuhaus HE, Henrichs G, Scheibe R. Starch degradation in intact amyloplasts isolated
Lytovchenko A, Bieberich K, Willmitzer L, Fernie AR. Carbon assimilation and metabolism from cauliower oral buds (Brassica oleracea L.). Planta 1995;195:496504.
in potato leaves decient in plastidial phosphoglucomutase. Planta 2002;215: Neuhaus HE, Husler RE, Sonnewald U. No need to shift the paradigm on the metabolic
80211. pathway to transitory starch in leaves. Trends Plant Sci 2005;10:1546.
Mangelsdorf PC. The genetics and morphology of some endosperm characters in maize. Niewiadomski P, Knappe S, Geimer S, Fischer K, Schulz B, Unte US, et al. The Arabidopsis
Conn Agric Exp Stn Bull 1926;279:509614. plastidic glucose 6-phosphate/phosphate translocator GPT1 is essential for pollen
Martnez-Barajas E, Delatte T, Schluepmann H, de Jong GJ, Somsen GW, Nunes C, et al. maturation and embryo sac development. Plant Cell 2005;17:76075.
Wheat grain development is characterized by remarkable trehalose 6-phosphate ac- Okazaki Y, Shimojima M, Sawada Y, Yoyooka K, Narisawa T, Mochida K, et al. A chloro-
cumulation pregrain lling: tissue distribution and relationship to SNF1-related pro- plastic UDP-glucose pyrophosphorylase from Arabidopsis is the committed enzyme
tein kinase 1 activity. Plant Physiol 2011;156:37381. for the rst step of sulfolipid biosynthesis. Plant Cell 2009;21:892909.
Massillon D, Bollen M, De Wulf H, Overloop K, Vanstapel F, Van Hecke P, et al. Demonstra- Olejnik K, Kraszewska E. Cloning and characterization of an Arabidopsis thaliana Nudix hy-
tion of a glycogen/glucose 1-phosphate cycle in hepatocytes from fasted rats. J Biol drolase homologous to the mammalian GFG protein. Biochim Biophys Acta
Chem 1995;270:193516. 2005;1752:13341.
McCoy JG, Arabshahi A, Bitto E, Bingman CA, Ruzicka FJ, Frey PA, et al. Structure and Olsson T, Thelander M, Ronne H. A novel type of chloroplast stromal hexokinase is the
mechanism of an ADP-glucose phosphorylase from Arabidopsis thaliana. Biochemistry major glucose-phosphorylating enzyme in the moss Physcomitrella patens. J Biol
2006;45:314562. Chem 2003;278:4443947.
McKibbin RS, Muttucumaru N, Paul MJ, Powers SJ, Burrell MM, Coates S, et al. Production Overlach S, Diekmann W, Raschke K. Phosphate translocator of isolated guard-cell chloro-
of high-starch, low glucose potatoes through over-expression of the metabolic regu- plasts from Pisum sativum L. transports glucose 6-phosphate. Plant Physiol 1993;101:
lator SnRK1. Plant Biotechnol J 2006;4:40918. 12017.
McLennan AG. The Nudix hydrolase superfamily. Cell Mol Life Sci 2006;63:12343. Patron NJ, Greber B, Fahy BF, Laurie DA, Parker ML, Denyer K. The lys5 mutations of barley
Meyer FD, Smidansky ED, Beecher BB, Greene TW, Giroux MJ. The maize Sh2r6hs reveal the nature and importance of plastidial ADP-glc transporters for starch synthe-
ADP-glucose pyrophosphorylase (AGP) large subunit confers enhanced AGP proper- sis in cereal endosperm. Plant Physiol 2004;135:208897.
ties in transgenic wheat (Triticum aestivum). Plant Sci 2004;167:899911. Periappuram C, Steinhaurer L, Barton DL, Taylor DC, Chatson B, Zou J. The plastidic phos-
Michalska J, Zauber H, Buchanan BB, Cejudo FJ, Geigenberger P. NTRC links built-in phoglucomutase from Arabidopsis. A reversible enzyme reaction with an important
thioredoxin to light and sucrose in regulating starch synthesis in chloroplasts. Proc role in metabolic control. Plant Physiol 2000;122:11939.
Natl Acad Sci U S A 2009;106:990813. Pontis HG, Babio JR, Salerno G. Reversible unidirectional inhibition of sucrose synthase ac-
Mikkelsen R, Mutenda KE, Mant A, Schrmann P, Blennow A. -Glucan, water dikinase tivity by disuldes. Proc Natl Acad Sci U S A 1981;78:66679.
(GWD): a plastidic enzyme with redox-regulated and coordinated catalytic activity Porchia AC, Curatti L, Salerno GL. Sucrose metabolism in cyanobacteria: sucrose synthase
and binding afnity. Proc Natl Acad Sci U S A 2005;102:178590. from Anabaena sp. strain PCC 7119 is remarkably different from the plant enzymes
Millar AH, Heazlewood JL. Genomic and proteomic analysis of mitochondrial carrier pro- with respect to substrate afnity and amino-terminal sequence. Planta 1999;210:3440.
teins in Arabidopsis. Plant Physiol 2003;131:44353. Pozueta-Romero J, Akazawa T. Biochemical mechanism of starch biosynthesis in amyloplasts
Miyagawa Y, Tamoi M, Shigeoka S. Overexpression of a cyanobacterial fructose-1,6-/ from cultured cells of sycamore (Acer pseudoplatanus). J Exp Bot 1993;44S:297306.
sedoheptulose-1,7-bisphosphatase in tobacco enhance photosynthesis and growth. Pozueta-Romero J, Ardila F, Akazawa T. ADP-glucose transport by the chloroplast adenyl-
Nat Biotechnol 2001;19:9659. ate translocator is linked to starch biosynthesis. Plant Physiol 1991a;97:156572.
Mhlmann T, Tjaden J, Henrichs G, Quick WP, Husler R, Neuhaus HE. ADPglucose drives Pozueta-Romero J, Frehner M, Viale A, Akazawa T. Direct transport of ADPglucose by an
starch synthesis in isolated maize endosperm amyloplasts. Characterization of starch adenylate translocator is linked to starch biosynthesis in amyloplasts. Proc Natl
synthesis and transport properties across the amyloplast envelope. Biochem J Acad Sci U S A 1991b;88:576973.
1997;324:5039. Pulido P, Spnola MC, Kirchsteiger K, Guinea M, Pascual MB, Sahrawy M, et al. Functional
Montero M, Eydallin G, Viale AM, Almagro G, Muoz FJ, Rahimpour M, et al. analysis of the pathways for 2-Cys peroxiredoxin reduction in Arabidopsis thaliana
Escherichia coli glycogen metabolism is controlled by the PhoPPhoQ regulatory chloroplasts. J Exp Bot 2010;14:404354.
system at submillimolar environmental Mg2 + concentrations, and is highly Purcell PC, Smith AM, Halford NG. Antisense expression of a sucrose non-fermenting-
interconnected with a wide variety of cellular processes. Biochem J 2009;424: 1-related protein kinase sequence in potato results in decreased expression of sucrose
12941. synthase in tubers and loss of sucrose-inducibility of sucrose synthase transcripts in
Montero M, Almagro G, Eydallin G, Viale AM, Muoz FJ, Bahaji A, et al. Escherichia coli leaves. Plant J 1998;14:195202.
glycogen genes are organized in a single glgBXCAP transcriptional unit possessing Ragel P. Identicacin y caracterizacin de los elementos implicados en el inicio de la
an alternative suboperonic promoter within glgC that directs glgAP expression. sntesis de almidn en plantas [PhD. Thesis]; 2012.
Biochem J 2011;433:10718. Rahimpour M, Montero M, Almagro G, Viale AM, Sevilla A, Canovas M, et al. GlgS, previ-
Mooney BP. The second green revolution? Production of plant-based biodegradable ously described as a glycogen synthesis control protein, negatively regulates motility
plastics. Biochem J 2009;418:21932. and biolm formation in Escherichia coli. Biochem J 2013;452:55973.
Moreno-Bruna B, Baroja-Fernndez E, Muoz FJ, Bastarrica-Berasategui A, Zandueta- Ral JP, Bowerman AF, Li Z, Sirault X, Furbank R, Pritchard JR, et al. Down-regulation of glu-
Criado A, Rodrguez-Lpez M, et al. Adenosine diphosphate sugar pyrophosphatase can, water-dikinase activity in wheat endosperm increases vegetative biomass and
prevents glycogen biosynthesis in Escherichia coli. Proc Natl Acad Sci U S A 2001;98: yield. Plant Biotechnol J 2012;10:87182.
812832. Regierer B, Fernie AR, Springer F, Perez-Melis A, Leisse A, Koehl K, et al. Starch content and
Mller J, Mettbach U, Menzel D, Samaj J. Molecular dissection of endosomal compart- yield increase as a result of altering adenylate pools in transgenic plants. Nat
ments in plants. Plant Physiol 2007;145:293304. Biotechnol 2002;20:125660.
Mller-Rber BT, Sonnewald U, Willmitzer L. Inhibition of the ADP-glucose pyrophos- Rodrguez-Lpez M, Baroja-Fernndez E, Zandueta-Criado A, Pozueta-Romero J. Adeno-
phorylase in transgenic potatoes leads to sugar-storing tubers and inuences tuber sine diphosphate glucose pyrophosphatase: a plastidial phosphodiesterase that pre-
formation and expression of tuber storage protein genes. EMBO J 1992;11:122938. vents starch biosynthesis. Proc Natl Acad Sci U S A 2000;97:870510.
Muoz FJ, Baroja-Fernndez E, Morn-Zorzano MT, Viale AM, Etxeberria E, Alonso-Casajs Roldn I, Wattebled F, Mercedes Lucas M, Delvalle D, Planchot V, Jimenez S, et al. The phe-
N, et al. Sucrose synthase controls both intracellular ADPglucose levels and transitory notype of soluble starch synthase IV defective mutants of Arabidopsis thaliana sug-
starch biosynthesis in source leaves. Plant Cell Physiol 2005;46:136676. gests a novel function of elongation enzymes in the control of starch granule
Muoz FJ, Morn-Zorzano MT, Alonso-Casajs N, Baroja-Fernndez E, Etxeberria E, formation. Plant J 2007;49:492504.
Pozueta-Romero J. New enzymes, new pathways and alternative view on starch bio- Rolletschek H, Hijirezaei M-R, Wobus U, Weber H. Antisense-inhibition of ADP-glucose
synthesis in both photosynthetic and heterotrophic tissues of plants. Biocatal pyrophosphorylase in Vicia narbonensis seeds increases soluble sugars and leads to
Biotransform 2006;24:6376. higher water an nitrogen uptake. Planta 2002;214:95464.
Rolletschek H, Nguyen TH, Husler RE, Rutten T, Gbel C, Feussner I, et al. Antisense inhi- Szydlowski N, Ragel P, Raynaud S, Roldn I, Montero M, Lucas MM, et al. Starch granule
bition of the plastidial glucose-6-phosphate/phosphate translocator in Vicia seeds initiation in Arabidopsis requires the presence of either class IV or class III starch
shifts cellular differentiation and promotes protein storage. Plant J 2007;51:46884. synthase. Plant Cell 2009;21:244357.
Rontein D, Dieuaide-Noubhani M, Dufourc Erick J, Raymond P, Rolin D. The metabolic ar- Tacke M, Yang Y, Steup M. Multiplicity of soluble glucan-synthase activity in spinach
chitecture of plant cells. Stability of central metabolism and exibility of anabolic leaves: enzyme pattern and intracellular location. Planta 1991;185:2206.
pathway during the growth cycle of tomato cells. J Biol Chem 2002;277:4394860. Tang G-Q, Sturm A. Antisense repression of sucrose synthase in carrot (Daucus carota L.)
Ryu CM, Farag MA, Hu CH, Reddy MS, Wei HX, Pare PW, et al. Bacterial volatiles promote affects growth rather than sucrose partitioning. Plant Mol Biol 1999;41:46579.
growth in Arabidopsis. Proc Natl Acad Sci U S A 2003;100:492732. Tarkka MT, Piechulla B. Aromatic weapons: trufes attack plants by the production of vol-
Ryu CM, Farag MA, Hu CH, Reddy MS, Kloepper JV, Pare PW. Bacterial volatiles induce sys- atiles. New Phytol 2007;175:3813.
temic resistance in Arabidopsis. Plant Physiol 2004;134:101726. Tauberger E, Fernie AR, Emmermann M, Renz A, Kossmann J, Willmitzer L, et al. Antisense
Sakulsingharoj C, Choi S-B, Hwang S-K, Edwards GE, Bork J, Meyer CR, et al. Engineering inhibition of plastidial phosphoglucomutase provides compelling evidence that potato
starch biosynthesis for increasing rice seed weight: the role of the cytoplasmic tuber amyloplasts import carbon from the cytosol in the form of glucose-6-phosphate.
ADP-glucose pyrophosphorylase. Plant Sci 2004;167:132333. Plant J 2000;23:4353.
Samaj J, Read ND, Volkmann D, Menzel D, Baluska F. The endocytic network in plants. Thvenot C, Simond-Cte E, Reyss A, Manicacci D, Trouverie J, Le Guilloux M, et al. QTLs
Trends Cell Biol 2005;8:42533. for enzyme activities and soluble carbohydrates involved in starch accumulation dur-
Sanz-Barrio R, Corral-Martnez P, Ancin M, Segui-Simarro JM, Farrn I. Overexpression of ing grain lling in maize. J Exp Bot 2005;56:94558.
plastidial thioredoxin f leads to enhanced starch accumulation in tobacco leaves. Thormhlen I, Ruber J, von Roepenack-Lahaye E, Ehrlich S-M, Massot V, Hmmer C, et al.
Plant Biotechnol J 2013;19:61827. Inactivation of thioredoxin f1 leads to decreased light activation of ADP-glucose
Sauer N. Molecular physiology of higher plant sucrose transporters. FEBS Lett 2007;581: pyrophosphorylase and altered diurnal starch turnover in leaves of Arabidopsis
230917. plants. Plant Cell Environ 2013;36:1629.
Schluepmann H, Pellny T, van Dijken A, Smeekens S, Paul M. Trehalose 6-phosphate is in- Thornbjrnsen T, Villand P, Denyer K, Olsen O-A, Smith AM. Distinct isoforms of
dispensable for carbohydrate utilization and growth in Arabidopsis thaliana. Proc Natl ADPglucose pyrophosphorylase occur inside and outside the amyloplast in barley en-
Acad Sci U S A 2003;100:684954. dosperm. Plant J 1996a;10:24350.
Scoeld GN, Hirose T, Gaudron JA, Upadhyaya NM, Ohsugi R, Furbank RT. Antisense sup- Thornbjrnsen T, Villand P, Kleczkowski LA, Olsen OA. A single gene encodes two differ-
pression of the rice sucrose transporter gene, OsSUT1, leads to impaired grain lling ent transcripts of ADP-glucose pyrophosphorylase small subunit from barley
and germination but does not affect photosynthesis. Funct Plant Biol 2002;29:81526. (Hordeum vulgare). Biochem J 1996b;313:14954.
Scott P, Kruger NJ. Inuence of elevated fructose-2,6-bisphosphate levels on starch mobi- Tiessen A, Prescha K, Branscheid A, Palacios N, McKibbin R, Halford NG, et al. Evidence
lization in transgenic tobacco leaves in the dark. Plant Physiol 1995;108:156977. that SNF1-related kinase and hexokinase are involved in separate sugar signaling
Serrato AJ, Prez-Ruiz JM, Spnola MC, Cejudo FJ. A novel NADPH thioredoxin reductase, pathways modulating post-translational redox activation of ADP-glucose pyrophos-
localized in the chloroplast, which deciency causes hypersensitivity to abiotic stress phorylase in potato tubers. Plant J 2003;35:490500.
in Arabidopsis thaliana. J Biol Chem 2004;279:438217. Tiessen A, Nerlich A, Faix B, Hmmer C, Fox S, Trafford K, et al. Subcellular analysis of
Shannon JC, Pien F-M, Liu K-C. Nucleotides and nucleotide sugars in developing maize en- starch metabolism in developing barley seeds using a non-aqueous fractionation
dosperms. Synthesis of ADP-glucose in brittle 1. Plant Physiol 1996;110:83543. method. J Exp Bot 2012;63:207187.
Shannon JC, Pien F-M, Cao H, Liu K-C. Brittle-1, an adenylate translocator, facilitates trans- Tjaden J, Mhlmann T, Kampfenkel K, Henrichs G, Neuhaus HE. Altered plastidic
fer of extraplastidial synthesized ADP-glucose into amyloplasts of maize endosperms. ATP/ADP-transporter activity inuences potato (Solanum tuberosum L.) tuber mor-
Plant Physiol 1998;117:123552. phology, yield and composition of tuber starch. Plant J 1998;16:53140.
She K-C, Kusano H, Koizumi K, Yamakawa H, Hakata M, Imamura T, et al. A novel factor Trethewey RN, Geigenberger P, Riedel K, Hajirezaei M-R, Sonnewald U, Stitt M, et al. Com-
FLOURY ENDOSPERM2 is involved in regulation of rice grain size and starch quality. bined expression of glucokinase and invertase in potato tubers leads to a dramatic re-
Plant Cell 2010;22:328094. duction in starch accumulation and a stimulation of glycolysis. Plant J 1998;15:
Shewmaker CK, Boyer CD, Wiesenborn DP, Thompson DB, Boersig MR, Oakes JV, et al. Ex- 10918.
pression of Escherichia coli glycogen synthase in the tubers of transgenic potatoes (So- Trethewey RN, Fernie AR, Bachmann A, Fleischer-Notter H, Geigenberger P, Willmitzer L.
lanum tuberosum) results in a highly branched starch. Plant Physiol 1994;104: Expression of a bacterial sucrose phosphorylase in potato tubers results in a
115966. glucose-independent induction of glycolysis. Plant Cell Environ 2001;24:35765.
Silvius JE, Snyder FW. Comparative enzymic studies of sucrose metabolism in the taproots Tsai C, Nelson OE. Starch-decient maize mutant lacking adenosine diphosphate glucose
and brous roots of Beta vulgaris L. Plant Physiol 1979;64:10703. pyrophosphorylase activity. Science 1966;151:3413.
Sivitz AB, Reinders A, Ward JM. Analysis of the transport activity of barley sucrose trans- Tsai HL, Lue WL, Lu KJ, Hsieh MH, Wang SM, Chen J. Starch synthesis in Arabidopsis is
porter HvSUT1. Plant Cell Physiol 2005;46:166673. achieved by spatial cotranscription of core starch metabolism genes. Plant Physiol
Slattery CJ, Kavakli IH, Okita TW. Engineering starch for increased quantity and quality. 2009;151:158295.
Trends Plant Sci 2000;5:2918. Valerio C, Costa A, Marri L, Issakidis-Bourguet E, Pupillo P, Trost P, et al.
Smidansky ED, Clancy M, Meyer FD, Lanning SP, Blake NK, Talbert LE, et al. Enhanced Thioredoxin-regulated -amylase (BAM1) triggers diurnal starch degradation in
ADP-glucose pyrophosphorylase activity in wheat endosperm increases seed yield. guard cells, and in mesophyll cells under osmotic stress. J Exp Bot 2010;62:54555.
Proc Natl Acad Sci U S A 2002;99:17249. Vargas WA, Pontis HG, Salerno GL. New insights on sucrose metabolism: evidence for an
Smidansky ED, Martin JM, Hannah LC, Fischer AM, Giroux MJ. Seed yield and plant bio- active A/N-Inv in chloroplasts uncovers a novel component of the intracellular carbon
mass increases in rice are conferred by deregulation of endosperm ADP-glucose trafcking. Planta 2008;227:795807.
pyrophosphorylase. Planta 2003;216:65664. Ventriglia T, Kuhn ML, Ruiz MT, Ribeiro-Pedro M, Valverde F, Ballicora MA, et al. Two
Smith AM. Prospects for increasing starch and sucrose yields for bioethanol production. Arabidopsis ADP-glucose pyrophosphorylase large subunits (APL1 and APL2) are cat-
Plant J 2008;54:54658. alytic. Plant Physiol 2008;148:6576.
Sonnewald U, Basner A, Greve B, Steup M. A second L-type isozyme of potato glucan Vespermann A, Kai M, Piechulla B. Rhizobacterial volatiles affect the growth of fungi and
phosphorylase: cloning, antisense inhibition and expression analysis. Plant Mol Biol Arabidopsis thaliana. Appl Environ Microbiol 2007;73:563941.
1995;27:56776. Villand P, Kleczkowski LA. Is there an alternative pathway for starch biosynthesis in cereal
Sparla F, Costa A, Lo Schiavo F, Pupillo P, Trost P. Redox regulation of a novel seeds? Z Naturforsch 1994;49:2159.
plastid-targeted -amylase of Arabidopsis. Plant Physiol 2006;141:84050. Viola R, Roberts AG, Haupt S, Gazzani S, Hancock RD, Marmiroli N, et al. Tuberization in
Splivallo R, Fischer U, Gbel C, Feussner I, Karlovsky P. Trufe volatiles inhibit growth and potato involves a switch from apoplastic to symplastic phloem unloading. Plant Cell
induce an oxidative burst in Arabidopsis thaliana. New Phytol 2007;175:41724. 2001;13:38598.
Stark DM, Timmerman KP, Barry GF, Preiss J, Kishore GM. Regulation of the amount of Walters RG, Ibrahim DG, Horton P, Kruger NJ. A mutant of Arabidopsis lacking the
starch in plant tissues by ADPglucose pyrophosphorylase. Science 1992;258:28792. triose-phosphate/phosphate translocator reveals metabolic regulation of starch
Stitt M, Heldt HW. Simultaneous synthesis and degradation of starch in spinach chloro- breakdown in the light. Plant Physiol 2004;135:891906.
plasts in the light. Biochim Biophys Acta 1981;638:111. Wang S-M, Lue W-L, Yu T-S, Long J-H, Wang C-N, Eimert K, et al. Characterization of
Stitt M, Zeeman S. Starch turnover: pathways, regulation and role in growth. Curr Opin ADG1, an Arabidopsis locus encoding for ADPG pyrophosphorylase small subunit,
Plant Biol 2012;15:111. demonstrates that the presence of the small subunit is required for large subunit sta-
Strand A, Zrenner R, Trevanion S, Stitt M, Gustafsson P, Gardestrm P. Decreased expres- bility. Plant J 1998;13:6370.
sion of two key enzymes in the sucrose biosynthesis pathway, cytosolic Wang A-Y, Kao M-H, Yang W-H, Sayion Y, Liu L-F, Lee P-D, et al. Differentially and devel-
fructose-1,6-bisphosphatase and sucrose phosphate synthase, has remarkably differ- opmentally regulated expression of three rice sucrose synthase genes. Plant Cell
ent consequences for photosynthetic carbon metabolism in transgenic Arabidopsis Physiol 1999;40:8007.
thaliana. Plant J 2000;23:75970. Wang Z, Chen X, Wang J, Liu T, Liu Y, Zhao L, et al. Increasing maize seed weight by en-
Streb S, Zeeman SC. Starch metabolism in Arabidopsis. The Arabidopsis book, 10. 2012. hancing the cytoplasmic ADP-glucose pyrophosphorylase activity in transgenic
p. e0160. http://dx.doi.org/10.1199/tab.0160. maize plants. Plant Cell Tissue Organ 2007;88:8392.
Streb S, Egli B, Eicke S, Zeeman SC. The debate on the pathway of starch synthesis: a closer Wang Y, Yu B, Zhao J, Guo J, Li Y, Han S, et al. Autophagy contributes to leaf starch degra-
look at low-starch mutants lacking plastidial phosphoglucomutase supports the dation. Plant Cell 2013;25:138399.
chloroplast-localised pathway. Plant Physiol 2009;151:176972. Weber H, Heim U, Borisjuk L, Wobus U. Cell-type specic, coordinate expression of two
Sulpice R, Pyl E-T, Ishihara H, Trenkamp S, Steinfath M, Witucka-Wall H, et al. Starch as a ADP-glucose pyrophosphorylase genes in relation to starch biosynthesis during
major integrator in the regulation of plant growth. Proc Natl Acad Sci U S A 2009;106: seed development of Vicia faba. Planta 1995;195:35261.
1034853. Weber H, Rolletschek H, Heim U, Golombek S, Gubatz S, Wobus U. Antisense-inhibition of
Sweetlove LJ, Burrell MM, ap Rees T. Starch metabolism in tubers of transgenic potato ADP-glucose pyrophosphorylase in developing seeds of Vicia narbonensis moderately
(Solanum tuberosum) with increased ADPglucose pyrophosphorylase. Biochem J decreases starch but increases protein content and affects seed maturation. Plant J
1996;320:4938. 2000;24:3343.
Weichert N, Saalbach I, Weichert H, Kohl S, Erban A, Kopka J, et al. Increasing sucrose up- Zhang H, Xie X, Kim M-S, Kornyeyev DA, Holaday S, Par PW. Soil bacteria augment
take capacity of wheat grains stimulates storage protein synthesis. Plant Physiol Arabidopsis photosynthesis by decreasing glucose sensing and abscisic acid levels in
2010;152:698710. planta. Plant J 2008;56:26473.
Weschke W, Panitz R, Sauer N, Wang Q, Neubohn B, Weber H, et al. Sucrose transport into Zhang H, Sun Y, Xie X, Kim M-S, Dowd SE, Par PW. A soil bacterium regulates plant ac-
barley seeds: molecular characterization of two transporters and implications for quisition of iron via deciency-inducible mechanisms. Plant J 2009;58:56877.
seed development and starch accumulation. Plant J 2000;21:45567. Zhang Y, Primavesi LF, Jhurreea D, Andralojc PJ, Mitchell RA, Powers SJ, et al. Inhibition of
Yu T-S, Lue W-L, Wang S-M, Chen J. Mutation of Arabidopsis plastid phosphoglucose isom- SNF1-related protein kinase 1 activity and regulation of metabolic pathways by
erase affects leaf starch synthesis and oral initiation. Plant Physiol 2000;123:31925. trehalose-6-phosphate. Plant Physiol 2009;149:186071.
Zeeman SC, Thorneycroft D, Schupp N, Chapple A, Weck M, Dunstan H, et al. Plastidial Zhou R, Cheng L. Competitive inhibition of phosphoglucose isomerase of apple leaves by
-glucan phosphorylase is not required for starch degradation in Arabidopsis leaves sorbitol 6-phosphate. J Plant Physiol 2008;165:90310.
but has a role in the tolerance of abiotic stress. Plant Physiol 2004;135:84958. Zrenner R, Willmitzer L, Sonnewald U. Analysis of the expression of potato
Zervosen A, Rmer U, Elling L. Application of recombinant sucrose synthase-large scale uridinediphosphate-glucose pyrophosphorylase and its inhibition by antisense RNA.
synthesis of ADP-glucose. J Mol Catal 1998;5:258. Planta 1993;190:24752.
Zhang L, Husler RE, Greiten C, Hajirezaei M-R, Haferkamp I, Neuhaus HE, et al. Overriding Zrenner R, Salanoubat M, Willmitzer L, Sonnewald U. Evidence of the crucial role of sucrose
the co-limiting import of carbon and energy into tuber amyloplasts increases the starch synthase for sink strength using transgenic potato plants (Solanum tuberosum L.). Plant
content and yield of transgenic potato plants. Plant Biotechnol J 2008;6:45364. J 1995;7:97107.

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JBA-06696; No of Pages 20

Biotechnology Advances xxx (2013) xxxxxx

Contents lists available at SciVerse ScienceDirect

Starch biosynthesis, its regulation and biotechnological approaches to improve

Calvin- Triose-P Triose-P

Calvin- Triose-P Triose-P

Calvin- Triose-P Triose-P OPPP

Fructose ADP Carriers/Endocytosis

PPi PPi UDP

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