Plant Science 298 (2020) 110572

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Plant Science
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Review article

Review: Crucial role of inorganic pyrophosphate in integrating carbon T

metabolism from sucrose breakdown to starch synthesis in rice endosperm
Sang-Kyu Lee, Jong-Seong Jeon*
Graduate School of Biotechnology and Crop Biotech Institute, Kyung Hee University, Yongin 17104, South Korea

A R T I C LE I N FO A B S T R A C T

Keywords: The endosperm is a primary constituent of mature seeds in rice as well as in other cereal crops, serving as the
Hypoxia major storage reserve of starch. Observations indicate that the central part of the endosperm is subject to hypoxic
Carbon metabolism conditions, which require a switch of energy metabolism owing to limited mitochondrial respiration. Uniquely,
Cereal this endosperm generates a large source of inorganic pyrophosphate (PPi) as a byproduct of the reaction of ADP
Endosperm
glucose pyrophosphorylase in the cytosol. Recent results derived from examination of the mutants of cereal
Inorganic pyrophosphate
crops, especially rice, for PPi-utilizing enzymes clearly suggest an important role of PPi as an alternative energy
Rice
Starch currency for integrating carbon metabolism from sucrose breakdown to starch synthesis in the endosperm. Thus,
Sucrose the present review provides an outline of the interlaced PPi-dependent metabolic pathways, which are critical
for starch synthesis in the endosperm in terms of energy metabolism, along with its application to enhance yield
potential.

1. Introduction
Rice (Oryza sativa) is a staple food crop for half of the world’s po-
pulation and, as one of the most important cereal crops, has become a
model monocot plant because of its relatively small genome, completed
genome sequence, and a large amount of available molecular and ge-
netic resources. The endosperm is a primary component of mature plant
seeds and is an important organ for the storage of food reserves, mostly
in the form of starch. It is therefore valuable to elucidate the mechanism
of starch synthesis in the endosperm [1].
Recent studies have provided detailed information on the cellular
and morphological features of developing rice seed [2,3]. Through
coenocytic nuclear division in the first two days after pollination (DAP),
and cellularization in three to five DAP, storage products accumulate
concurrently with the differentiation of aleurone and starchy en-
dosperm cells from six to 20 DAP. Starch synthesis initiates from the
central region of the endosperm and continues gradually towards the
peripheral region. At approximately 21 DAP, the endosperm enters the
desiccation mature phase until 30 DAP. This phase is characterized by
tightly coordinated developmental and metabolic changes, mainly re-
lated to the accumulation of starch, but a comprehensive understanding
of the underlying mechanisms is still lacking.
Although relatively less detailed information is available for rice,
the endosperms of other examined cereal crops, such as wheat, barley,
and maize, which similarly accumulate a large amount of starch as their
main reserve, but only a little in embryos, are commonly exposed to
hypoxic conditions, i.e. limited oxygen supply [4–8]. In cereal seeds,
the pericarp, which is a distinct layer, develops the chloroplasts that
drive photosynthesis, which can contribute to oxygenation. In contrast,
the central region of cereal endosperm is highly hypoxic (for example,
falling to < 5 μM oxygen in barley endosperm), unlike in the photo-
synthetically active pericarp where hypoxia does not occur [8]. At
night, hypoxia increases in the central region of cereal endosperm
(< 2 μM oxygen in barley endosperm). Thus, during the starch synth-
esis phase, the reduced level of oxygen in the endosperm causes a de-
crease in cellular energy status. Due to the overall similar seed struc-
tures and physiology of rice and the aforementioned cereal crops, the
hypoxic condition in the central region of the endosperm is most likely
a universal feature among these crop species. The localized difference
between the central and peripheral environments of the endosperm
may require a clear switch from central carbon metabolism [glycolysis
and the tricarboxylic acid (TCA) cycle] to alcoholic fermentation within
the central endosperm, as demonstrated in a dynamic proteomic ana-
lysis of developing rice endosperms [9].
A supply of oxygen is necessary for sufficient production of the
energy currency adenosine triphosphate (ATP) from mitochondrial re-
spiration through central carbon metabolism in heterotrophic organs,
such as roots and seeds, in which photosynthesis is not active. In

⁎
Corresponding author.
E-mail address: jjeon@khu.ac.kr (J.-S. Jeon).

https://doi.org/10.1016/j.plantsci.2020.110572
Received 10 April 2020; Received in revised form 28 May 2020; Accepted 19 June 2020
Available online 25 June 2020
0168-9452/ © 2020 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license
(http://creativecommons.org/licenses/BY/4.0/).
S.-K. Lee and J.-S. Jeon Plant Science 298 (2020) 110572

Fig. 1. Outlines of the starch synthetic pathway from sucrose breakdown in the central endosperm region, under hypoxic conditions, in rice. Cereal crops possess a
cytosolic form of ADG-Glc pyrophosphorylase (AGP) that mainly functions in producing ADP-Glc and inorganic pyrophosphate (PPi) in the endosperms. ADP-Glc is
imported to the amyloplast by the ADP-Glc transporter, BT1, via counter-exchange with ADP, and used for starch synthesis in the organelle. PPi is consumed by UDP-
Glc pyrophosphorylase (UGP) coupled with sucrose synthase (SUS) to generate Glc-1-P, which stimulates the production of ADP-Glc via the cytosolic activity of AGP.
ATP is supplied from the glycolysis pathway via PPi-dependent fructose-6-phosphate 1-phosphotransferase (PFP) and pyruvate, orthophosphate dikinase (PPDK) by
recycling NAD+ from a noncyclic mode of the TCA cycle via cytosolic Alanine aminotransferase (cAlaAT) and Malate dehydrogenase (cMDH). PPi and its utilizing
key enzymes are indicated in red and blue, respectively (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of
this article).
AGP = ADP-Glc pyrophosphorylase; BT1 = ADP-glucose transporter; ADP-Glc = ADP-glucose; PFK = ATP-dependent phosphofructokinase; cAlaAT = cytosolic
alanine aminotransferase; cAspAT = cytosolic aspartate aminotransferase; cwINV = cell wall invertase; cMDH = cytosolic malate dehydrogenase; NIN = neutral
invertase; cPPDK = cytosolic pyruvate, orthophosphate dikinase; F1,6 P = fructose-1-6-bisphosphate; F6 P = fructose-6-phosphate; G1 P = glucose-1-phosphate; G6
P = glucose-6-phosphate; HXK = hexokinase; mMDH = mitochondrial malate dehydrogenase; PEP = phosphoenolpyruvate; PFP = PPi-dependent fructose-6-
phosphate 1-phosphotransferase; PK = pyruvate kinase; SUS = sucrose synthase; UDP-Glc = UDP-glucose; UGP = UDP-Glc pyrophosphorylase.

particular, to rapidly accumulate a large amount of starch, the cereal
endosperm demands a considerable amount of ATP in producing ADP-
glucose (ADP-Glc), the direct precursor of starch synthesis. However,
low ATP production due to the hypoxic conditions in the endosperm
cannot sufficiently meet the demand of ATP-consuming cellular meta-
bolic processes [8]. As a potential strategy to cope with this energy
crisis, the induction of inorganic pyrophosphate (PPi)-utilizing enzyme
reactions has been proposed for cells and tissues enduring hypoxic
conditions [10–12]. PPi is produced as a byproduct of biosynthetic
pathways in the cytosol and also in many other subcellular organelle
compartments [13,14]. Uniquely, the endosperm in cereal crops, as
well as in grasses, generates a large source of PPi from the ADP glucose
pyrophosphorylase (AGP) reaction in the cytosol, which can drive PPi-
utilizing pathways (see Section 2) (Fig. 1).
There have recently been a number of functional studies on the
endosperm mutants for PPi-utilizing enzymes in rice and also in other
cereal crops (Table 1). However, some of these reports focused only on
the molecular and genetic roles of the enzymes but neglected their
crucial roles in hypoxic endosperm conditions. Thus, the present review
revisits those previous studies and proposes that PPi plays an important
role, as an alternative of ATP, in integrating carbon metabolism from
sucrose breakdown to starch synthesis in the endosperm.
2. Supply of a source of PPi by cytosolic ADP glucose
pyrophosphorylase
It is widely known that AGP plays a key role in starch synthesis. AGP
catalyzes the important regulatory step that produces ADP-Glc and PPi
using glucose-1-phosphate (Glc-1-P) and ATP. The resulting ADP-Glc
serves as a glucosyl donor during α-1,4-glucan synthesis (Fig. 1). While
prokaryotic AGPs are homotetramers composed of four identical sub-
units [15], in higher plants AGPs exist as heterotetramers consisting of
each two large (LSU) and small (SSU) subunits [16–18]. AGPs are
present in the plastids of all plant species, but some isoforms function
uniquely in the endospermic cytosol of cereal crops, in addition to
grasses [19–22], with only one cytosolic LSU and SSU isoform evolving
in each species. Interestingly, one of the major SSU genes produces two
distinct isoforms harboring different first exons, one of which possesses
a transit peptide that directs it to leaf chloroplasts, and the other in the
endospermic cytosol [22–25]. It is evident in rice and other cereal crops
that all of the cytosolic AGP null mutants, including shrunken2 or
brittle2, contain low levels of starch in their endosperm because of low
AGP activity (Table 1). This assertion is well supported by the fact that
the mutations of the ADP-Glc transporter, which transports ADP-Glc
from the cytosol into plastids, cause low starch levels in the endosperm
in brittle1 (Fig. 1) [26–29]. Despite the importance of starch synthesis in
the endosperm, the reason for the cytosolic production of ADP-Glc is
rarely discussed.
In general, one ATP is required to produce one ADP-Glc from Glc-1-
P. Given that the endosperm is exposed to hypoxic conditions in rice
and other cereal crops, ATP may not be sufficiently supplied through
central carbon metabolism in the absence of aerobic respiration. The
activity of one of the TCA cycle enzymes, citrate synthase, is decreased
by 85 %, leading to a 60 % reduction in respiration between six to 10
DAP in wheat endosperm [30]; this is consistent with a decrease in the
density of cristae in mitochondria between four to 16 DAP in wheat
endosperm [31]. The reduction in mitochondrial respiration is also
corroborated by the phenotype of cytosolic AGP mutants in rice, which

2
S.-K. Lee and J.-S. Jeon
Table 1
Functional studies of the genes involved in PPi-dependent energy metabolism of starch biosynthesis in cereal endosperms.
Enzyme Species Accession or locus number
cytosolic AGP SSU Oryza sativa LOC_Os08g25734
Hordeum vulgare P55238
Zea mays AAK69627
cytosolic AGP LSU Oryza sativa LOC_Os01g44220
SUS Zea mays GRMZM2G089713
UGP Oryza sativa LOC_Os02g02560
PFP1 Oryza sativa LOC_Os06g13810
cytosolic PPDK Oryza sativa LOC_Os05g33570
Zea mays GRMZM2G306345
cytosolic AlaAT Oryza sativa LOC_Os10g25130
Hordeum vulgare Z26322.1
cytosolic MDH Oryza sativa LOC_Os10g33800
V-PPase Oryza sativa LOC_Os05g06480
AGP SSU = ADP glucose pyrophosphorylase small subunit; AGP LSU = ADP glucose pyrophosphorylase large subunit; SUS = sucrose synthase; UGP = UDP-Glc
pyrophosphorylase; PEP = phosphoenolpyruvate; PPDK = pyruvate, orthophosphate dikinase; AlaAT = alanine aminotransferase; MDH = malate dehydrogenase;
V-PPase = PPi-dependent vacuolar proton pump.
accumulate near normal levels of starch in the peripheral endosperm
region that can diffuse oxygen from the surface, but almost no starch in
the central interior region of the endosperm (Table 1) [8,22]. These
findings suggest an important role of cytosolic AGP isoforms in the
endosperm at low oxygen level; that is, in the hypoxic endosperm
conditions that limit ATP production, the supply of PPi by the cytosolic
AGP can be an alternative energy source to drive starch synthesis.
3. PPi-utilizing UDP-glucose pyrophosphorylase coupled with
sucrose synthase
Starch synthesis in the endosperm relies on a supply of sucrose
transported from photosynthetic leaf tissues. The site of phloem un-
loading in rice occurs in the vascular system on the ventral side of the
ovary, where sucrose is cleaved by cell wall invertase (cwINV) prior to
being transported into the endosperm (Fig. 1). The function of invertase
(INV) is particularly advantageous in the early stages of endosperm
development during cellularization and cell expansion [32,33]. As the
endosperm matures to the starch synthesis phase, a switch to sucrose
synthase (SUS)-dependent sucrose cleavage is observed as an adapta-
tion to the hypoxic conditions [34]. SUS-dependent sucrose cleavage
requires one ATP, reducing the ATP cost in the hypoxic endosperms,
while INV-dependent sucrose cleavage demands two ATPs to phos-
phorylate two hexoses. Consistently, hypoxic environments repress the
expression of INV and up-regulate SUS expression [35,36]. However,
mutations in the rice Grain-filling gene1 and the maize Miniature1 for
cwINV, and the maize Shrunken1 for SUS, are still capable of activating
starch synthesis (Table 1), suggesting a clear redundancy between these
two sucrose cleavage pathways in the endosperm.
During the starch accumulation phase of the endosperm under hy-
poxic conditions, PPi-utilizing UDP-glucose pyrophosphorylase (UGP)
functions actively in catalyzing the formation of Glc-1-P, a substrate for
AGP reaction in starch synthesis, from the UDP-glucose (UDP-Glc)
produced in the SUS-dependent sucrose cleavage (Fig. 1). UGP has
around a 40-fold higher activity than SUS in cereal endosperms, in-
cluding those of rice, wheat, and maize, under hypoxic conditions [37].
The UGP reaction coupled with SUS to produce Glc-1-P may be ensured
by the supply of sufficient PPi released from cytosolic AGP activity,
which stimulates cytosolic ADP-Glc formation. Thus, the concerted
metabolic process of SUS-UGP-AGP that ensures the effective use of PPi
is greatly advantageous for starch synthesis in the hypoxic endosperm
(Fig. 1). Mutations of a rice UGP isoform, floury endosperm8, have been
shown to exhibit a chalky opaque endosperm because of impaired
starch synthesis (Table 1), revealing its important role in starch
synthesis [38,39]. Notably, when levels of PPi become low in the
Plant Science 298 (2020) 110572
Mutant phenotype Reference
Shrunken endosperm in null mutant (osagps2) [22]
Shrunken endosperm in null mutant (Risø16) [23]
Shrunken endosperm in null mutant (Brittle2) [25]
Shrunken endosperm in null mutant (osagpl2) [22]
Shrunken endosperm in null mutant (Shrunken) [34]
Opaque endosperm in null mutant (floury endosperm 8) [38,39]
Opaque endosperm in null mutant (pfp1) [44,45]
Opaque endosperm in null mutant (floury endosperm 4) [48,49]
Opaque endosperm in null mutant (pdk1, pdk2) [50]
Opaque endosperm in null mutant (osalaat1)/ Increased seed weight in overexpression line [54,55]
Increased seed weight in overexpression line [65]
Opaque endosperm in null mutant (floury endosperm 16)/ Increased seed weight in [63]
overexpression line
Chalky opaque endosperm in Zhenshan97 carrying high expression QTL (Chalk5) [64]
cytosol, the amount of UDP-Glc appears to increase in Arabidopsis
seedlings [40]. This finding may suggest a reversible UGP function that
can balance the direction of metabolic processes towards either supply
or consumption of PPi to adjust AGP-Glc formation. It is also worth
considering that SUS can directly produce ADP-Glc at a cost of ATP for
starch synthesis [41].
4. PPi-driven glycolysis
In hypoxic endosperm conditions, the limitation of ATP production
favors PPi-driven glycolysis to generate ATP. Among a number of en-
zymes involved in glycolysis, two PPi-utilizing enzymes, the mutants of
which have been well characterized, are PPi-dependent fructose-6-
phosphate 1-phosphotransferase (PFP; also called PPi-dependent
phosphofructokinase) and pyruvate, orthophosphate dikinase (PPDK).
These enzymes are highly expressed in cereal endosperms.
Interestingly, some energy-poor anaerobic microorganisms, such as the
bacterium Priopionibacterium shermanii and the parasitic amoeba
Entamoeba histolytica, possess only PFP and PPDK, instead of ATP-de-
pendent phosphofructokinase (PFK) and pyruvate kinase (PK), respec-
tively [42]. This suggests a potential role of these PPi-utilizing enzymes
in energy-saving metabolism under oxygen deficiency. Using ATP, PFK
catalyzes the irreversible reaction to produce fructose-1-6-bisphosphate
(F-1,6-P) from fructose-6-phosphate (F-6-P). Instead of ATP, PFP in-
terconverts F-6-P to F-1,6-P by using PPi. PFP activity is highly in-
creased in anoxically germinating rice coleoptiles [10]. The opaque
endosperm phenotype in the maize bZIP transcription factor mutant
opaque2 is alleviated by upregulating PFP in the endosperm [43]. The
rice PFP mutants, pfp1-1, pfp1-2 and pfp1-3 have been shown to exhibit
opaque endosperms (Table 1) [44,45]. Given that PFP activity increases
under low oxygen conditions, the limitation of ATP may induce PPi-
utilizing PFP activity to reduce the cost of ATP in the endosperm.
PK is a key enzyme that produces pyruvate and ATP using phos-
phoenolpyruvate (PEP) and ADP in glycolysis. The biological function
of PK has been studied little due to the presence of many isoforms in
plants [46]. PPDK, an enzyme similar to PK, catalyzes a reversible re-
action to produce pyruvate and ATP from PEP, AMP, and PPi (Fig. 1).
Rice, has two PPDK genes, one encoding OsPPDKA and another pro-
ducing two transcripts of chloroplastic PPDK (chOsPPDKB) and cyto-
solic PPDK (cyOsPPDKB) [47]. Among them, mutations of the cytosolic
form of OsPPDKB, which is predominant in rice endosperm, are char-
acterized by opaque endosperm (Table 1) [9,48,49]. Although there is
no direct evidence that the endosperm-preferential cytosolic PPDK is
induced under hypoxic conditions, it clearly functions as a key enzyme
in the PPi-dependent glycolytic pathway. An opaque seed phenotype is
3
S.-K. Lee and J.-S. Jeon
also conditioned by PPDK deficiency in maize [50]. OsPPDKA is upre-
gulated in germinating rice seedlings under oxygen-deficiency condi-
tions, but its function in endosperm is unknown [51].
5. Alternative pyruvate-consuming metabolic flux using cytosolic
alanine aminotransferase and malate dehydrogenase
Glycolysis is activated to increase ATP level via the PPi-dependent
PFP and PPDK in hypoxic endosperm conditions. This process leads to a
high production of pyruvate, but the activity of the TCA cycle remains
low under hypoxic conditions [8,52]. As an alternative process, the
accumulated pyruvate is metabolized by a cytosolic alanine amino-
transferase (cAlaAT) in cereal endosperm cells. cAlaAT catalyzes a re-
versible conversion reaction of glutamate and pyruvate to alanine and
2-oxoglutarate. Expression of cAlaAT is induced at the starch accumu-
lation phase in hypoxic endosperm conditions [53–55]. Rapid produc-
tion of alanine by cAlaAT keeps glycolysis active through the con-
sumption of pyruvate (Fig. 1). Thus, cAlaAT functions as a key enzyme
that can determine the direction of the metabolic flux in the endosperm
[8]. Mutants of alanine aminotransferase 1 (OsAlaAT1), a cytosolic
isoform that is the most highly expressed among the five AlaAT genes in
rice endosperm, have been shown to exhibit opaque endosperm
(Table 1) because of altered starch structure [54,55]. The resulting 2-
oxoglutarate and glutamate are further metabolized to aspartate and
oxaloacetate by aspartate aminotransferase (AspAT), which is highly
expressed in the endosperm [56].
An increase in fermentation activity in hypoxic endosperms is im-
portant for maintaining glycolysis by supplying NAD+ to sustain ATP at
a level that is adequate for starch synthesis [9]. In addition to the fer-
mentation process, a metabolite profiling analysis of the hypoxic cen-
tral part of barley endosperm revealed an important pyruvate-con-
suming process that is a non-cyclic mode of the TCA cycle via cytosolic
isoforms of cAlaAT and malate dehydrogenase (cMDH), in addition to
mitochondrial MDH (mMDH) [8]. cMDH reversibly catalyzes the re-
duction of oxaloacetate to malate while oxidizing NAD(P)H to NAD
(P)+. In general, cMDH is known as a key component of ‘malate valves’,
which act as systems for balancing the ATP/NAD(P)H ratio by recycling
malate between the cytosol and organelles, including mitochondria
[57]. Malate is highly accumulated in cereal endosperms [58–60]. It
may be possible that oxaloacetate is synthesized by phosphoenolpyr-
uvate carboxylase (PEPC) spending CO2, which then converts to malate
in the cytosol. However, this assumption is not reliable because of the
much higher activity of cMDH compared to PEPC activity [58,59].
Thus, it is most likely that pyruvate is converted into oxaloacetate by
the concerted action of cAlaAT and AspAT, and further converted to
malate by cMDH to supply the electron acceptors needed to produce
ATP from glycolysis. In this process, cMDH drives the reaction from
oxaloacetate and NADH to malate and NAD+ than the reverse reaction
in the endosperm under energy-deficient hypoxic conditions (Fig. 1)
[8,61]. A reduction of cytosolic MDH activity under oxidizing condi-
tions supports this observation [62], and mutations in cytosolic MDH
have been shown to result in severely opaque endosperms (Table 1)
[63], revealing the important role of the pyruvate-consuming metabolic
flux via cAlaAT and cMDH in cereal endosperms.
6. Conclusion and future prospects
Although the starch of cereal endosperms accounts for most of the
carbohydrates that humans consume, we are still far from fully un-
derstanding carbon metabolism. The functions of some amyloplastic
enzymes in starch synthesis, including starch synthases, starch
branching enzymes, and starch debranching enzymes, are relatively
well known. However, the most important question regarding the
supply of energy required for synthesizing a pool of ADP-Glc, the direct
precursor and building block of starch, has not been clearly answered in
regard to the hypoxic endosperm conditions in rice as well as in other
4
Plant Science 298 (2020) 110572
cereal crops. Recently, direct evidence of the importance of PPi-uti-
lizing enzymes for starch synthesis in the endosperm has come from
examination of their genetic mutants. The results presented herein
provide solid evidence regarding the crucial roles of the PPi-dependent
interlaced metabolic flux of SUS-UGP-AGP, and the PPi-dependent
glycolysis via PFP and cPPDK coupled with the non-cyclic mode of the
TCA cycle via cAlaAT and cMDH, both of which sustain glycolytic flux
and thus adequate levels of ATP in the endosperm (Fig. 1). Thus, in
terms of energy metabolism, impaired ADP-Glc synthesis can affect the
supply of PPi, the alternative metabolic energy supply, which may lead
to an inactivation of these entire PPi-dependent energy metabolic
processes. To further support the crucial role of the proposed PPi-uti-
lizing energy metabolism in cereal endosperms, the other enzymes of
this pathway whose endosperm mutants have not yet been character-
ized need to be examined. One of these additional enzymes is fructose-
6-phosphate-2-kinase/fructose-2,6-bisphosphatase, which can regulate
the first important glycolysis-committed step to F-1,6-P by producing
F2,6-P.
It is worth noting the possible function of the vacuolar PPi-depen-
dent proton pump (V-PPase), which can replace the vacuolar ATP-de-
pendent proton pump (V-ATPase). Under low oxygen conditions, the V-
PPase can be important to prevent a rapid decrease in the cytosolic pH
when H+-ATPase is inhibited due to ATP deficiency. The V-PPase is
strongly induced in oxygen-deficient rice tissues [51]. Interestingly,
elevated activity of the V-PPase, coding for a dominant Chalk5 allele,
causes endospermic opacity in rice, possibly by disturbing the pH
homeostasis in the endosperm, which is coupled with a great increase in
small vesicle-like structures, thus forming air spaces among endosperm
storage substances and resulting in a chalky endosperm (Table 1) [64].
However, it is debatable whether this unexpected increase of V-PPase
may lead to the depletion of PPi in the cytosol and disturb starch
synthesis that leads to the mutant endosperm (Fig. 1). Whether or not
its contribution in energy metabolism is critical remains unanswered.
Opaque endosperm in rice and other cereal crops is a major problem
readily influenced by environmental factors, such as temperature and
field fertility, and is an undesirable trait for human food that negatively
affects milling, cooking, and eating. A number of endosperm mutants
involved in the PPi-dependent metabolic fluxes typically exhibited
opacity, suggesting an important role of these metabolic fluxes for
normal endosperm development under stress conditions. Another in-
teresting observation in the context of biotechnology is that over-
expression of some of these PPi-utilizing enzymes increases starch
synthesis and seed weight. Thus, the constitutive activation of the PPi-
dependent pathways may provide an effective way to enhance yield
potential, as already demonstrated in AlaAT [55,65] and MDH [63]
transgenic lines (Table 1), as well as in SUS- or AGP-overexpressing
transgenic lines [66,67]
Lastly, it is worth noting that programmed cell death (PCD) occurs
simultaneously with the rapid accumulation of starch from the central
part to the peripheral region of rice endosperm [3]. Most likely, PCD in
rice endosperm occurs in a non-lytic manner that nuclei and other or-
ganelles remain functional during this process [68,69], which may
allow starch synthesis to occur in a large endosperm compartment with
a shared cytoplasm. Thus, it would be interesting to investigate a
contribution of PPi-dependent metabolic pathways for starch synthesis
during PCD process in rice endosperm.
Declaration of Competing Interests
All of the authors have read the manuscript and declare no conflict
of interest.
Acknowledgments
This work was supported by grants from the Next Generation
S.-K. Lee and J.-S. Jeon
BioGreen 21 program (Project No. PJ013172012019), the Rural
Development Administration, and the Mid-Career Researcher program
(Project No. 2020R1A2C2012976), National Research Foundation of
Korea.
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