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Biosynthesis of fatty oils in higher plants

Article  in  Russian Journal of Plant Physiology · January 2014

DOI: 10.1134/S1021443714010130

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ISSN 10214437, Russian Journal of Plant Physiology, 2014, Vol. 61, No. 1, pp. 1–18. © Pleiades Publishing, Ltd., 2014.
Original Russian Text © R.A. Sidorov, V.D. Tsydendambaev, 2014, published in Fiziologiya Rastenii, 2014, Vol. 61, No. 1, pp. 3–22.

REVIEWS

Biosynthesis of Fatty Oils in Higher Plants

R. A. Sidorov and V. D. Tsydendambaev
Timiryazev Institute of Plant Physiology, Russian Academy of Science, Botanicheskaya ul. 35, Moscow, 127276 Russia;
fax: 8 (499) 9778018; email: vdt@ippras.ru
Received April 22, 2013

Abstract—Triacylglycerols (TAGs) are one of the most important storage compounds of higher plants; they
are the basis for essentially all plant oils and are used by the cell as a reserve source of energy and carbon.
Therefore, as a rule, plants store oils almost exclusively in their seeds. Plant oils are not only a major food and
feed products, but also the raw material for obtaining many nonfood products from drying oils and lubricants
to biofuel. TAGs differ from other storage compounds in that in the course of fruit ripening their quantitative
and qualitative composition does not remain constant, but undergoes significant changes. Therefore, the bio
synthesis pathways of TAGs in living organisms have been actively studied for the past several decades, and
today enough data can be presented to outline as how and where these processes occur in the plant cell. The
present review is devoted to a brief description of current ideas about the ways and mechanisms of TAG for
mation and accumulation in higher plants.

Keywords: higher plants, fruits, seeds, maturation, biosynthesis, fatty acids, TAG, acDAG, oleosomes, plas
tids, endoplasmic reticulum
DOI: 10.1134/S1021443714010130

INTRODUCTION valuable ones (such as oil palm, avocado, olive, and

Higher plants accumulate during ontogeny a vari
ety of substances: proteins, carbohydrates (glucose,
fructose, sucrose, starch, etc.), lipids (fatty oils),
essential and mustard oils, and many other products of
secondary metabolism. This plant capability is widely
used by humans, using the storage organs of plants in
food or animal feed and as raw materials for various
products (medicines, alcohols, drying oils, varnishes,
lubricants, industrial oils, etc.).
Plant oils are the most important food in the
human diet providing a significant part of the energy
needs of his organism and serving as the source of
essential fatty acids (FAs). The base of essentially all
fatty oils is triacylglycerols (TAGs), the esters of tri
atomic alcohol glycerol, all hydroxyl groups of which
are esterified by FAs. Storage of oils by plants occurs
almost exclusively in the seeds, where TAGs are a
reserve of energy and carbon [1, 2]. At the same time,
there are a number of plants, including commercially
Abbreviations: ACP—acylcarrier protein; acDAG—3acetyl
1,2diacylsnglycerol; DAG—1,2diacylsnglycerol; DGAT—
1,2diacylsnglycerol acyltransferase; ER—endoplasmic reticu
lum; FA—fatty acid; G3P—snglycero3phosphate; GPAT—
snglycerol3phosphate acyltransferase; GPDH—snglycero3
phosphate dehydrogenase; KAS—ketoacyl synthase; LPA—
lysophosphatidic acid; LPAAT—lysophosphatidic acid acyl
transferase; NAG—neutral acylglycerol; PA—phosphatidic
acid; PC—phosphatidylcholine; PE—phosphatidylethanol
amine; PDAT—phosholipid: diacylglycerol acyltransferase;
POM—plants with oily mesocarp; PPA—phosphatase of phos
phatidic acid; PUFA—polyunsaturated FA; TAG—1,2,3tria
cylsnglycerol.
sea buckthorn), in which a significant amount of oil is
contained not only in the seeds, but also in the juicy
outside seed fruit parts (mesocarp, pericarp, hypanth
ium, arillus, etc.), where, in addition to oils, other
nutrients (carbohydrates, vitamins, and various sec
ondary compounds) accumulate. Such plants are
called as “plants with oily mesocarp” (POM). Oils in
the outside seed fruit parts make such fruits attractive
to animals, especially birds, by which plants can
spread efficiently [3]. Although these oils are accumu
lated in different parts of the same fruit, they always
sharply differ in the composition of their constituent
FAs [4–8]. In particular, the juicy arils of these fruits
contain usually highly saturated oil with the predomi
nance of С16 FAs, palmitic and hexadecenoic and/or
octadecenoic acids, whereas the seeds of the same
fruits accumulate TAGs rich in polyunsaturated С18
FAs, linoleic and αlinolenic. Finally, there is a group
of plants that accumulate in the seeds and arils the
TAG analogs, neutral acylglycerols (NAGs) differing
in their composition and chemical nature (see below).
Taking into account such drastic differences in accu
mulated oils, it can be assumed that the biosynthetic
pathways of both TAG and other NAG in different
plant tissues and organs are different, and therefore
many studies are devoted to their studying.
Typically, all of hydroxyl groups of the glycerol
moiety in the TAG molecule are esterified by FA resi
dues. When in the sn1,3 and sn2 positions, acyls of
different FAs are present, the molecule of this TAG is

1
2 SIDOROV, TSYDENDAMBAEV
asymmetric and exhibits optical activity; however, in
the case of usual FAs, differences in the TAG molecule
polarizability are negligible, whereas TAGs with
unusual FAs exhibit the high optical activity [9]. Thus,
the high optical activity determined by acyls of
unusual FAs in sn3 position was observed in TAGs
from Mannina emarginata seeds containing coriolic
(13hydroxycis9,trans11octadecadienoic) acid [10].
Along with various FAs, TAGs of some plant oils
may include nonlipid functional groups. Thus, in the
seeds of euonymus (Euonymus verrucosus), Himalayan
balsam (Impatiens roylei), and I. edgeworthii, unusual
optically active 3acetylsn1,2diacylglycerols
(acDAGs) containing the residue of acetic acid in the
sn3 position were found [11, 12]. TAGs from the
seeds of plants from the genus Dracunulus contain
unusual ωphenylalkanoic and ωphenylalkenoic
acids; they comprise up to 13% of total FAs in TAGs
[13]. In the lipophilic excretions from buds of poplar
(Populus lasiocarpa), widely spread in China, TAGs
containing residues of phenylcarbonic acids, mainly
sn1,3dipcoumaroyl2acetylglycerols, sn1,3di
caffeyl2acetylglycerols, and sn1pcoumaroyl3
caffeyl2acetylglycerols were detected [14]. In the
stem bark of Aquilaria malaccensis (Thymelaeaceae),
spread in India, Myanmar, Borneo, Thailand, and the
Philippines, 1,3dibehenyl2ferruloylglycerols [15]
were detected. In Lilium longiflorum plants, unusual
DAG analogs, sn1 and sn3diacetyl2glucosylg
lycerols, with acetic acid residues instead FAs in sn1
and sn3 positions and glucose in sn2 position were
found [16].
FATTY ACIDS
Fatty acids (FAs) are the main components of
TAGs largely determining oil properties. As a rule,
they are divided into major, minor, and unusual [17].
TAGs and membrane lipids often comprise lauric
(12:0), myristic (14:0), palmitic (16:0), stearic (18:0),
oleic (Δ918:1), linoleic (Δ9,1218:2), and αlino
lenic (Δ9,12,1518:3) acids. These seven FAs with
even number of carbon atoms in the aliphatic chain
are widespread in nature and are the main lipid com
ponents. They are called major or usual [17, 18]. At the
same time, the qualitative composition of FAs in
TAGs and membrane lipids in many plants can differ
substantially: TAGs often contain FAs, which are
absent from membrane lipids (over 1000 individual FA
species are detected only in TAGs). The reasons for
this will be considered by us below.
The acids homologous to major FAs, but present in
lower amounts in lipids (no more than 5% of total
FAs) are called minor. Minor FAs also include
homologs of the major FAs with the greater or fewer
number of carbon atoms or differing in the position or
configuration of double bonds. Major and minor FAs
are present in the lipids of most plant species; there
fore, they are called as usual FAs. Other FAs differing
RUSSIAN JOURNAL OF PLANT PHYSIOLOGY
in double bond configuration or position or in the
presence of coupled double or triple bonds and also
met in the plant lipids belong to unusual FAs. Over
1000 different unusual FAs are found in the seed oils;
however, it is supposed that plants may comprise sev
eral thousands of unusual FAs [18, 19]. As a rule,
unusual FA chains contain from 8 to 30 carbon atoms,
double bonds in unusual positions of the molecule,
and one or several diverse functional groups, for exam
ple, methyl, hydroxyl, epoxyl, cyclopropane, cyclo
pentane, etc. [17, 18]. Unusual FAs can comprise up
to 90% of total FAs in oil.
Meanwhile, the attribution of a FA to the group of
major, minor, or unusual FAs is rather arbitrary and
may be used only in a comparative study of the total
lipids in a large number of objects. In each case, for
each class of lipids FA attribution to one or another
group depends on its percentage in the given lipid class
and on the frequency of its occurrence in studied lipids
of this plant material. Thus, lauric and myristic acids,
usually belonging to major FAs, are found in lipids of
most plants only in small amounts, but in tropical
plant oils their proportion may exceed 40% [20].
Biosynthesis of Fatty Acids in Higher Plants
The biosynthesis of FAs occurs predominantly in
the two subcellular compartments, viz. chloroplasts
and endoplasmic reticulum (ER). De novo synthesis
from acetylCoA of such FAs as palmitic or stearic
and also desaturation of stearic acid to oleic one
occur in plastids, whereas the further conversion of
oleic acid in linoleic and further linolenic acid occurs
in the ER [21].
The synthesis of FAs from acetylCoA starts in
plastids (Fig. 1) and occurs in three stages. Firstly, the
irreversible carboxylation of acetylCoA is catalyzed
by acetylCoA carboxylase with the formation of
malonylCoA. Then the malonyl group is transferred
to the acylcarrier protein (ACP) and, due to the oper
ation of plastid acetylCoA carboxylase complex,
malonylACP is formed, which is a primary substrate
of FA synthetase [22]; the formation of malonylCoA
is an obligatory stage of FA synthesis [17].
During the second stage, successive condensation
of newly formed malonylCoA with growing, bound to
ACP acyl chains occurs. In such a way, by the succes
sive addition of twocarbon fragments to the growing
chain, palmitoylACP is produced [22]. Four reac
tions run during each cycle of condensation. Firstly,
due to the condensation of acetylCoA and malo
nylCoA, 3ketobutylACP is formed catalyzed by
ketoacyl synthase III (KAS III); then it is reduced to
3hydroxyacylACP, dehydrated to enoylACP, and
reduced to butyrylACP. Subsequent cycles of con
densation of acylACP with malonylCoA are cata
lyzed by KAS I and proceed to the formation of myris
toylACP and palmitoylACP [23]. The elongation of
palmitoylACP to stearoylACP and its desaturation
Vol. 61 No. 1 2014
 BIOSYNTHESIS OF FATTY OILS IN HIGHER PLANTS 3

AcetylCoA carboxylase MCMT

AcetylCoA MalonylCoA MalonylACP
СО2 ACP CoA
AcetylCoA
KAS III

HADH KAR
EnoylACP 3HydroxyacylACP 3KetobutylACP

ENR
KAS I
ButyrylACP MiristoylACP
MalonylACP
5 MalonylACP KAS I

KAS II
StearoylACP PalmitoylACP

FatB MalonylACP FatB

Δ9AcylACP desaturase
C18:0 C16:0
FatA Plastid
OleoylACP С18:1

Cytosol

Fig. 1. FA biosynthesis in higher plants.

FatA and FatB—acylACP thioesterases hydrolyzing predominantly oleoylACP and diverse saturated and unsaturated acyl
ACP, respectively; ENR—enoylACP reductase; KAR—ketoacylACP reductase; KAS—ketoacyl synthase; MCMT—
malonylCoA : ACPmalonyl transferase; HADH—hydroxyacylACP dehydrase.

by Δ9desaturase to oleoylACP are catalyzed by
KAS II and occur also in plastids.
During the third stage of FA biosynthesis, palmi
toylACP, stearoylACP, and oleoylACP are exported
into the cytosol and included into acylCoA and acyl
lipid pools [1]. In some plants, the process of elonga
tion can continue, and in this case socalled “FAs with
very long chains” (VLCFAs) including 20 and more
carbon atoms are formed [17, 22].
ACPthioesterases (enzymes belonging to the class
of acylACP hydrolases) catalyze the termination of
elongation; they hydrolyze acylACP with the forma
tion of free FA, which can cross the plastid membrane
to be reactivated outside the organelle [24]. Two main
types of plant thioesterases are described: FatA prefer
ably cleaving oleate from ACP and FatB hydrolyzing
diverse saturated and unsaturated acylACPs. In some
plant species, thioesterases are found, which are spe
cific to acylACPs with shortchain acyls [25, 26]. The
interaction between FA synthetase, Δ9desaturase, and
the two types of thioesterases determines the ratio
between FAs produced in plastids, which will be further
used by the cell for the synthesis of glycerolipids [19].
After export from plastids, oleic acid enters into the
cytosolic pool and is imported into the ER in the form
of CoA ester. Then, oleic acid becomes available for
further conversion into linoleic and further αlino
lenic acid; substratespecific desaturases are involved
in these reactions [21, 22, 27]. At the same time, other
modifications of FA acyls may occur in the ER. It was
shown that in castor (Ricinus communis) plants oleic
acid was subjected to hydroxylation by oleate hydrox
ylase (FAH12) with the formation of unusual ricino
leic (12hydroxyoleic) acid [27]. As a result, some FAs
synthesized in plastids and ER are included in the
structural components of cell membranes (e.g., phos
pholipids or galactolipids) and the rest are included in
the TAGs.

RUSSIAN JOURNAL OF PLANT PHYSIOLOGY Vol. 61 No. 1 2014

4 SIDOROV, TSYDENDAMBAEV
Unusual Monounsaturated FAs
The synthesis of most monounsaturated FAs is cat
alyzed by soluble plastid desaturase, which normally
inserts a double bond between the 9th and 10th carbon
atoms of the acyl residue of stearoylACP [18]. At the
same time, some plants capable of the synthesis of
unusual monounsaturated FAs have additional desat
urases, which structure is close to that of Δ9desatu
rase but despite this fact they introduce the ethylene
bond in another position of the acyl and can manifest
a different substrate specificity [19]. Thus, the seeds of
some plants from the family Apiaceae (Umbelliferae),
carrot and coriander in particular, contain oils
enriched in petroselinic acid (Δ618:1). This unusual
FA is formed due to the operation of plastid Δ4desat
urase introducing the double bond between the 4th
and 5th carbon atoms of the acyl in palmitoylACP,
conversing it into Δ4hexadecenoylACP, which then
elongates to petroselinic acid [28].
MECHANISMS OF TAG BIOSYNTHESIS
IN HIGHER PLANTS
Triacylglycerols are present in all eukaryotes: ani
mals, plants, fungi, and protistas [17], and also in
some prokaryotes [29]. TAG biosynthesis in all organ
isms is characterized by common metabolic pathways,
which can be traced from bacteria to humans [30].
TAGs are the most important storage components
of seeds in many plants, including oilseeds, such as
sunflower (Helianthus annuus), rapeseed (Brassica
napus), soybean (Glycine max), etc. [31, 32]. At the
same time, such plants as the olive (Olea europaea),
avocado (Persea americana), durian (Durio zibethi
nus), oil palm (Elaeis guineensis), sea buckthorn (Hip
pophaё rhamnoides), and some others, accumulate
TAGs not only in the seeds, but also in juicy fruit parts
outside the seeds (pericarp, hypanthium, etc.) [4–8].
TAG accumulation in the seeds occurs during fruit
development; in their cells TAGs accumulates in the
oily inclusions (oleosomes) and are stored until the
start of germination; during germination TAGs can be
used as a source of energy produced in the process of
βoxidation, or as a source of acetylCoA, or as a sub
strate in the reaction of acylation, for example, during
the synthesis of phospholipids [17, 30].
In plant seeds, glycerolipids can be synthesized in
two similar pathways, which are known as prokaryotic
and eukaryotic systems. These pathways are localized
in different subcellular compartments and are charac
terized by a similar twostage enzymatic conversion of
snglycerol3phosphate (G3P) into phosphatidic
acid (PA), but different further conversions of PA into
structural, storage, or signaling lipids [33]. Despite the
closeness of these two biosynthetic pathways, enzymes
that catalyze acylation reactions are unique to each
system.
RUSSIAN JOURNAL OF PLANT PHYSIOLOGY
Enzymes of the prokaryotic pathway are localized
in plastids, whereas enzymes of the eukaryotic path
way – in the cytosol and ER. In the prokaryotic path
way, FA residues are directly transferred from ACP to
G3P; whereas in the eukaryotic pathway, FAs are
removed from ACP by acylACP thioesterases and
released free FAs than are transported in the cyto
plasm, where they are converted into acylCoA. Dur
ing the synthesis of membrane and storage lipids [10],
acyl groups are used in the ER by acyltransferases of
the eukaryotic G3P pathway [10]. In dependence on
subcellular localization, these enzymes may differ in
their structure, thus forming independent clusters in
phylogenetic investigations [34].
The accumulation of storage lipids in plants
includes de novo FA biosynthesis in the plastid stroma
with their subsequent involvement in the glyceride
metabolism resulting in TAG formation in the ER [1].
These two pathways of TAG biosynthesis were called
as acylCoAdependent and acylCoAindependent.
AcylCoADependent Pathway of TAG Biosynthesis
in Plants
In the acylCoAdependent pathway of TAG bio
synthesis (Fig. 2), which is also known as the Kennedy
pathway, acylCoA is a substrate for successive reac
tions of glycerol residue acylation; as a result, sn1,2
diacylglycerols (DAGs) are produced with the help of
DAG acyltransferases (DGAT) [35]. Glycerol3
phosphate (G3P) serves as a source of the glycerol res
idue; G3P is produced from one of the glycolysis
products, dihydroxyacetone, because of catalytic
action of snglycerol3phosphate dehydrogenase
(GPDH) [24].
The Kennedy pathway starts from G3P acylation
by snglycerol3phosphate acyltransferase (GPAT)
with the formation of lysophosphatidic acid (LPA).
The second acylCoAdependent acylation is cata
lyzed by lysophosphatidic acid acyltransferase
(LPAAT) with the formation of PA. Then, phos
phatase of PA releases phosphate groups from PA with
the formation of DAGs [1]. Finally, DGAT converses
DAGs into TAGs, using acylCoA as a donor of the
acyl residue [22].
Glycerol3Phosphate Acyltransferase Role
in TAG Biosynthesis
GPAT is a membranebound enzyme initiating the
process of TAG synthesis by the transfer of acyl from
acylCoA or acylACP to the sn1 position of G3P,
producing LPA [19]. In some plants, the two forms of
GPAT were found, which are localized in plastid and
cytoplasmic cell compartments. This enzyme may
manifest a high specificity toward the substrate, for
example, to the residues of oleic acid as a donor of
acyls, or does not manifest such specificity, using any
acyl, including that of saturated FAs. Plants can use
Vol. 61 No. 1 2014
 BIOSYNTHESIS OF FATTY OILS IN HIGHER PLANTS
R1COCoA
COH COCOR1
HOC HOC
GPAT
COP COP
Glycerol3phosphate Lysophosphatidic
acid
COCOR1
R2COOC
COCOR3
Triacylglycerol
Fig. 2. AcylCoAdependent pathway of TAG biosynthesis (Kennedy pathway) [35].
LPAAT—acyltransferase of lysophosphatidic acid; GPAT—glycerol3phosphate acyltransferase; DGAT—diacylglycerol
acyltransferase; PPA—phosphatase of phosphatidic acid.
this different substrate specificity of GPAT toward res
idues of saturated and unsaturated FAs in their stress
responses to a sharp environmental temperature low
ering [36].
The Role of Lysophosphatidic Acid Acyltransferase
in TAG Biosynthesis
This enzyme catalyzes transfer of FA acyl from
acylCoA into sn2 position of LPA with the forma
tion of PA. It was shown for the developing endosperm
of oil palm, which TAGs are rich in lauric acid, that
there are LPAAT forms specific to acyls of saturated
FAs [37]. The activity of oleoyllysophosphatidate
LPAAT was constant during endosperm development
despite the fact that the activity of lauroylphosphati
date LPAAT was detected only at the stages of lauric
acid synthesis [38]. This observation allowed a sugges
tion that specialized genes exist encoding LPAAT dif
fering from analogous “housekeeping genes” [19].
Moreover, microsomes of developing embryos of cigar
flower (Cuphea lanceolata), which oil contains up to
90% of capric acid, under in vitro conditions synthe
sized efficiently dicaprioyldiacylglycerols; this allows
a suggestion that different plants comprise different
LPAAT, including those specific to acyls with the
medium chain length [39]. This is supported by other
studies as well. Thus, in the seeds of higherucic rape
seed cultivars and some other plant species from the
family of Cruciferae (Brassicaceae), LPAAT discrimi
nates eicosenoyl, erucoyl, and nervonoylacylCoA
RUSSIAN JOURNAL OF PLANT PHYSIOLOGY Vol. 61
5
R2COCoA
COCOR1
R2COOC
LPAAT
COP
Phosphatidic
acid
PPA
R3COCoA
COCOR1
R2COOC
DGAT
COH
1,2Diacylglycerol
[24]. In contrast, LPAAT from plants of Tropae
olaceae and Limnanthaceae families manifest a high
selectivity to acylCoA with acyls of VLCFAs [40].
The activity of LPAAT in plants is connected with
numerous membrane systems, including chloroplasts,
ER, and inner mitochondrial membrane, and this
determines the occurrence of several different forms of
this enzyme [41]. In plants, LPAAT is selective toward
the residues of unsaturated FAs [19], but this enzyme
can also discriminate acyl groups with the shorter or
longer aliphatic chains. During seed development in
some genetically modified plants, LPAAT specific to
unusual acyls can insert such acyls into the sn2 posi
tion of TAGs [42].
Phosphatase of Phosphatidic Acid
The cytoplasmic enzyme phosphatase of phospha
tidic acid (PPA) dephosphorylates PAs with the for
mation of DAGs [43]. However, not all DAGs are used
by the cell for TAG biosynthesis – some of them
become substrates for the synthesis of membrane
phospholipids [44]. In plants there are two types of
PPA involved in the synthesis of glycerolipids, which
are present in soluble and membranebound states.
These enzymes are also involved in the metabolism of
phospholipids, in particular, in processes of their deg
radation. It was shown that PA accumulates in plants
transiently in response to various stresses. Dephos
phorylation by PPA reduces PA signaling activity by its
conversion into DAG. This mechanism may play an
No. 1 2014
6 SIDOROV, TSYDENDAMBAEV
important role in the development of stress response
related to the changes in the composition of mem
brane lipids [45].
In plant cells, two forms of PPA were detected. One
of them, PPA1, is a Mg2+dependent enzyme, whereas
another one, PPA2, is independent of magnesium cat
ions and can participate in the utilization of some
phosphatecontaining substrates, including PA [46].
Although PPA1 is present in the cell in the soluble
form but after oleate binding, as it was shown on devel
oping safflower cotyledons, it can be incorporated in
the membrane of the ER [24].
Diacylglycerol Acyltransferase
DGAT, an enzyme of the Kennedy pathway, cata
lyzes the third acylCoAdependent acylation, result
ing in DAG conversion into TAG. DGAT is a mem
branebound enzyme localized in the ER but also
present in oleosomes and plastids [47]. Despite the
fact that most DGAT of the first and second types pre
dominate in the cell, plants contain other types of
enzymes with DGAT activity different in their struc
ture [48, 49]. Among them are soluble DGAT of pea
nut [50], wax ester synthetase/acetyldiacylglycerol
acyltransferase (WSD1) of arabidopsis [32], and
DGAT specific for acyls of acetic acid of the winged
tree (euonymus) [51]. DGAT substrate specificity
depends on several factors, such as the FA composi
tion of the DAG pool, the concentration of acylCoA,
and temperature [1].
The gene encoding DGAT1 was cloned from
mouse DNA on the basis of this enzyme homology
with genes encoding acylCoA : cholesterase of mam
mals [52]. Several DGAT homologs of plants and
fungi were cloned and characterized [53, 54].
The second family of genes encoding DGAT2,
which nucleotide sequences differ from those of
DGAT1 genes, was firstly found in the fungus Mortier
ella ramanniana [55]. Some other DGAT2 genes of
animals, fungi, and plants were cloned and character
ized [55, 56]. It was experimentally demonstrated that
in some plants the expression of genes encoding
DGAT2 was correlated with the accumulation in their
seeds of TAGs containing FAs with conjugated double
bonds or hydroxyl groups [32, 57]. The role of DGAT1
and DGAT2 in TAG biosynthesis and oil accumula
tion evidently depends on plant species. It is estab
lished that, in plants with TAGs containing only usual
FAs, the leading enzyme involved in oil accumulation
in the seeds is DGAT1, whereas DGAT2 is responsible
for oil accumulation in the seeds of plants, in which
TAGs unusual FAs predominate [32, 40, 57].
The third variety of DGAT, a soluble DGAT3, was
recently found in peanut and some other plants [49,
57]; however, we know about it very little so far. Acyl
CoAdependent acyltransferase called wax esters/dia
cylglycerol acyltransferase (WS/DGAT) was discov
ered and isolated from the bacterium Acinetobacter sp.
RUSSIAN JOURNAL OF PLANT PHYSIOLOGY
This enzyme can use fatty alcohols and DAGs as acyl
acceptors in the synthesis of wax esters and TAGs,
respectively [40, 58]. A large number of genes homol
ogous to a gene encoding a DGAT3 of Acinetobacter sp.
was found in arabidopsis [59].
It turned out that DGAT could be one of the limit
ing factors during storage lipid accumulation in plants
[40, 60] and thus plays an important role in the forma
tion of qualitative and quantitative TAG composition.
It was shown that overexpression of genes encoding
DGAT1 resulted in the enhanced oil accumulation in
the arabidopsis seeds, whereas the inhibition of these
genes resulted in a decrease in the seed oil content
[60]. It should be added that DGAT activity is possibly
important also for correct acyl distribution of unusual
FAs in seed storage lipids [49].
AcylCoAIndependent TAG Biosynthesis
As an alternative to the Kennedy pathway, FAs
arising de novo may be firstly integrated into the lipids
of plastid membrane and/or those of the ER and are
stored as TAGs only later (Fig. 3) [22].
Newly synthesized FAs may be directly incorpo
rated into phosphatidylcholines (PC), bypassing PA
and intermediate DAGs [61]. Acyls of FAs from PC
may be included into TAGs via the PC reverse conver
sion into DAG or through the functioning of phos
pholipid:diacylglycerol acyltransferase (PDAT). This
enzyme belongs to the family of lecitine:cholesterol
acyltransferases [59], which catalyze TAG formation
by acyl transfer from sn2 position of the phospholipid
molecule to DAG with phosphatidylethanolamine
(PE), a preferable donor of FA acyls in yeast and
plants [62]. It was shown that arabidopsis PDAT could
use not only PE but also other phospholipids as donors
of FA acyls; it could accept diverse acyl groups with
the chain length from 10 to 22 carbon atoms [32]. For
example, in yeast PDAT1 is a main enzyme involved
in TAG accumulation during culture exponential
growth [63].
The activity of genes encoding PDAT (especially in
the seeds of oily plants) maybe is a factor determining
the exclusion of unusual FAs from membrane phos
pholipids and their transport into TAGs [1]. It was
suggested that the regulation of genes encoding PDAT
might play an important role in the maintenance of
the constant composition of membrane lipids due to
their reduction or due to the removal of DAGs, the
effector molecules of the phosphatidylinosite signaling
pathway, which is actively involved in the formation of
plant response to stress [59].
In another case of acylCoAindependent trans
acylation, DAG transacylase catalyzes acyl transfer
between two DAG molecules with the formation of
TAG and monoacylglycerol (MAG) [64]. This pro
cess can run in the reverse direction and be used for
TAG remodeling [40]. However, genes encoding this
enzyme are not identified until now.
Vol. 61 No. 1 2014
 BIOSYNTHESIS OF FATTY OILS IN HIGHER PLANTS
There is growing evidence of the existence of one
more alternate acylindependent pathways of TAG
formation in plants using the enzyme choline phos
photransferase or aminoalcohol phosphotransferase.
Choline phosphotransferase can catalyze TAG forma
tion through PC conversion into DAG, which can be
later acylated by DGAT or PDAT. It has been sug
gested that permanent reversible DAG transformation
into PC can control, in particular, the composition of
polyunsaturated fatty acids (PUFA) in the seed oil
[65], making thereby choline phosphotransferase by
the enzyme controlling the pathway, in which PUFA
become available for inclusion in the TAGs [27, 65].
Finally, it has been recently demonstrated that
phospholipases (in particular, phospholipase D) may
play an important role in PC conversion into TAGs
[66]. Due to the suppression of phospholipase D activ
ity by RNA interference, the dilinoleylPC and dilino
leylPE accumulation in soybean seeds at concentra
tions much higher than those observed in wildtype
plants was shown.
REGULATION OF TAG BIOSYNTHESIS
AND OIL ACCUMULATION IN PLANTS
The accumulation of large amounts of storage sub
stances occurs during seed maturation and is regulated
mainly by genetic program controlling the develop
ment of these organs [67]. The regulation of oil pro
duction and accumulation occurs at many levels [68].
Some enzymes involved in the biosynthesis, accumu
lation, and degradation of neutral lipids were identi
fied and characterized, and it was found a large num
ber of their isoforms and varieties. In most plant spe
cies tested, proteins involved in the metabolism of
neutral lipids are rather conservative and have a high
degree of homology to each other [30].
The synthesis and accumulation of storage compo
nents are regulated by numerous transcription factors
involved in the complex network of genetic programs,
hormonal and metabolic signaling systems [3, 69].
Some of these transcription factors are known as the
main regulators because of their pronounced ability to
influence the activity of other factors [24]. The most
important regulators of seed maturation and storage
material accumulation include genes from families
LEAFY COTYLEDON (LEC1 and LEC2), FUSCA3
(FUS3), ABSCISIC ACID INSENSITIVE3 (ABI3),
and WRINKLED1 (WRI1) [24, 69]. Among the
known factors that regulate gene activity related to the
storage of reserve substances, gene WRI1 is responsi
ble for the regulation of oil accumulation [70, 71]. The
gene WRI1 encodes transcription factors from the
family APETALA2ethylenedependent element
binding proteins [70]. This factor is also responsible
for specific regulatory activity of other transcription
factors, such as LEC2 and maybe LEC1 during FA
biosynthesis [3, 72]. In particular, it is established that
WRI1 regulates the expression of genes encoding the
RUSSIAN JOURNAL OF PLANT PHYSIOLOGY Vol. 61
7
enzymes of glycolysis, during which sucrose is con
verted into precursors of TAG biosynthesis [70]. In
wri1 mutants, oil accumulation in the seeds was by
80% lower than in wildtype plants [22].
Other transcription factors are also involved in the
regulation of lipid metabolism in maturing seeds.
Thus, it was shown that overexpression of LEC1 gene
in arabidopsis seeds exceeded the levels of the expres
sion of known genes encoding enzymes of FA metab
olism almost by 60%, whereas the amount of FAs
stored in the seed oil increased 4.7fold as compared
with wildtype plants [73]. In the same work, it was
shown that the involvement of LEC1 in the control of
FA biosynthesis depended partially on genes ABI3,
FUS3, and WRI1.
The FUS3 gene evidently also plays an important
role in oil accumulation in arabidopsis because the
amount of its transcripts increased together with the
increment of transcripts of genes involved in FA bio
synthesis [71]. It was also demonstrated that the
induced expression of FUS3 gene in arabidopsis
resulted in an intense expression of genes related to FA
biosynthesis in young plants. Similar results were
obtained for arabidopsis protoplasts transiently
expressing FUS3 gene [24].
It was shown that in soybean seeds DNAbinding
proteins with “zinc finger motif” (protein modules
interacting with DNA, RNA, other proteins and small
molecules) affect also oil accumulation. Overexpres
sion of GmDof4 and GmDof11 genes resulted in the
activation of genes encoding the Rsubunit of acetyl
CoA carboxylase and acylCoA FA synthetase,
respectively, also involved in FA biosynthesis. Evi
dently, this was a reason for the increase of lipid con
tent by11–24% in transgenic arabidopsis plants with
intense expression of GmDof4 and GmDof11 genes in
comparison with wildtype plants [24, 74]. Neverthe
less, along with FA biosynthesis control by transcrip
tion factors, other levels of regulation exist, for exam
ple, allosteric regulation of such enzymes as acetyl
CoA carboxylase [75] or plastid pyruvate kinase [22,
72] involved in the formation of important precursors
of TAG biosynthesis.
Accumulation of Unusual FAs within TAGs
It was already noted above that unusual FAs in
membrane lipids exert a negative effect on membranes
and hence on the cell as a whole, and to escape these
effects, the cell inserts these FAs into neutral lipids,
TAGs. Thus, along with storage function TAGs play an
additional role in “utilization” of unusual FAs. In fact,
in most known plant species synthesizing unusual
FAs, the proportion of these FAs does not exceed 5%,
whereas in their TAGs the proportion of unusual FAs
can attain up to 85% of total FAs [18]. What are the
mechanisms of unusual FA discrimination preventing
their inclusion in membrane lipids and facilitating
their incorporation in TAGs? This question is of
No. 1 2014
8 SIDOROV, TSYDENDAMBAEV
R2COOC
CPT
or
AAPT
D
as e
COCOR1
ip
pho l
R2COOC
Phos
COH
1,2Diacylglycerol
DGAT
COCOR1
R2COOC
COCOR3
Triacylglycerol
Fig. 3. AcylCoAindependent pathway of TAG biosynthesis.
AAPT—aminoalcohol phosphotransferase; DGAT—diacylglycerol acyltransferase; PDAT—phospholipid:diacylglycerol acyl
transferase; CPT—choline phosphotransferase.
importance for understanding the mechanisms of oil
accumulation with the high content of unusual FAs,
many of which are widely applied in medicine and
industry.
For today the reasons for the accumulation of
unusual FAs in TAGs is usually explained by the
hypothesis of substrate specificity of the TAG biosyn
thetic enzymes [18].
Substrate Specificity of Enzymes of TAG Biosynthesis
as the Factor of Unusual FA Discrimination
One of the ways of unusual FA incorporation into
TAGs preventing their entering into membrane lipids
is the mechanism of TAG biosynthesis with the partic
ipation of thioesterases. It is known that, at prokary
otic glycerolipid biosynthesis, the enzymes succes
sively acylating G3P need in acylACP as a substrate
(see above). Therefore, after hydrolysis by
thioesterases of acylACP containing unusual FA res
idue, the latter becomes unavailable for the further
RUSSIAN JOURNAL OF PLANT PHYSIOLOGY
COCOR1
COP (CH2)2N(CH3)3
Phosphatidyl choline
COCOR1
R2COOC
COP (CH2)2NH2
Phosphatidyl ethanolamine
PDAT
COCOR1
HOC
COP (CH2)2NH2
Lysophosphatidyl ethanolamine
Endoplasmic reticulum
synthesis of polar lipids but can be easily used for TAG
formation. This is the case, for example, in the genus
Cuphea, the oil of which accumulates many FAs with
a medium chain length (from 8 to 14 carbon atoms).
Another mechanism is also known: FAs with unusual
position of a double bond, such as, for example,
petroselinic acid (Δ618:1), are synthesized and ester
ify ACP, and in such a form this “unusual” acyl can
easily be incorporated in plastid membrane lipids [76].
However, this does not occur because the plants with
seeds enriched in petroselinic acid comprise the
thioesterase specific to this acyl, which does not per
mit its insertion into chloroplast membranes [77]. At
the same time, polar lipids of carrot and coriander
seeds contain rather high amount of unusual petrose
linic acid (from 10 to 20% vs. 70–75% in TAGs),
whereas in other plant species the proportion of
unusual FAs in membrane lipids only infrequently
exceeds 5% [18].
Vol. 61 No. 1 2014
 BIOSYNTHESIS OF FATTY OILS IN HIGHER PLANTS
Specificity of Acyltransferases as the Factor
of Discrimination of Unusual FAs during Glycerolipid
Biosynthesis
Acyltransferases are enzymes playing a key role in
the biosynthesis of both membrane and neutral lipids.
Numerous in vitro experiments showed that these
enzymes manifest the high selectivity to the activated
acylcontaining substrate and to the molecule, the
target of acyl group transfer. Thus, acyltransferase
selectivity may be a key factor determining the distri
bution of unusual FAs between membrane and neu
tral lipids [78].
TAG biosynthesis in plants from the genus Cuphea
is a good example of how this distribution occurs. All
three acyltransferases involved in TAG biosynthesis
can use as a substrate acyls of major FAs and FAs with
the medium chain length. Thus, the set of FAs in sn1
position depends on the composition of the acylCoA
pool [39]. At the same time, acyl groups inserted into
the sn1 position affect acylCoA specificity and selec
tivity of LPAAT, the enzyme acylating TAG in sn2
position. When C18 acid is present in the sn1 position
of the LPA residue, LPAAT uses for esterification of
sn2 position predominantly С18acyls, whereas when
the FA acyl with the medium chain length is in the sn
1 position, LPAAT uses for esterification of sn2 posi
tion the acyl of medium length. The mechanism of
such specificity is evidently related to the existence of
LPAAT isoforms [78]. As a result, PA molecules are
produced with different substitutions in sn1 and sn2
positions of the molecule. DGAT, the enzyme acylat
ing TAG in sn3 position, recognizes 10:0/10:0DAG
and esterified in sn3 position also by FA residues with
medium chain length [39, 79]. Thus, in plants from
the genus Cuphea, there is a characteristic set of acyl
transferases specific to FA acyls with medium chain
length; their operation results in the synthesis of TAGs
containing predominantly 10:0–14:0 FAs.
In vitro studies of other plant species containing
unusual FAs showed that their acyltransferases incor
porated unusual FAs into TAGs in a similar way [79].
A particularly important role is played here by DGAT
as their relationship with the pools of DAG and PC, as
well as high specificity to the FA composition of
DAGs leads to the fact that virtually no unusual FAs
are built into the polar lipids; they are completely
localized in the TAGs. As distinct from these DGAT,
DGAT of plants accumulating oil with usual FAs, for
example, sunflower and maize, do not manifest in
vitro any selectivity relative to unusual FAs [80].
“OILY INCLUSIONS”
Being synthesized in the ER, possibly, in its spe
cialized subdomains [57], TAGs are then transported
and accumulated in subcellular structures called “oily
bodies” or oleosomes [30]. Oleosomes are spherical
organelles from 0.2 to 25 μm in diameter, which con
RUSSIAN JOURNAL OF PLANT PHYSIOLOGY Vol. 61
9
tain the matrix, mainly from TAGs, surrounded by a
onelayer phospholipid membrane with aliphatic
chains directed inside (toward TAGs) and polar phos
phate groups directed outside, toward the cytosol [80].
“Oily inclusions” are found in essentially all types of
plant cells, and, despite the fact that the mechanisms
of oleosome formation in tissues outside the seeds is
not completely elucidated, it seems evident that they
participate in numerous unique processes, such as
stress responses, the development of tolerance to
pathogens, and hormone metabolism [82]. Oleosomes
are evidently formed in the ER microdomains; how
ever, this issue still continues to be debated [83]. In the
onelayer membrane of oleosomes, a specific protein
family was found; these proteins comprise from 1 to
4% of the oleosome weight [31, 84]. The pool of these
proteins, oleosins, was studied in detail and described
for several members of the Cruciferae (Brassicaceae)
plants [85].
Oleosins are most widespread proteins of oleo
somes. They can cover the whole surface of these
organelles, providing for organelle stability by means
of steric hindrance and electronegative repulsion;
therefore, oleosomes never fuse with each other and
do not form aggregates within the seeds [83, 86]. Along
with oleosins, other proteins (caleosins, sterolesins,
and aquaporins) were isolated from oleosome mem
branes. Some of them, together with oleosins, play a
crucial role in storage lipid mobilization and degrada
tion, occurring via βoxidation during seed germina
tion [87]. It is established that oleosins, at the end of
the molecule directed toward the cytosol, along with
N and Cdomains of binding, comprise the two
regions responsible for binding with polar groups of
phospholipids; caleosins contain calciumbinding site
(socalled EFhand motif), whereas sterolesins have
on their Cends a domain with dehydrogenase of
hydroxylated steroids (GSDdomain) [88]. In their
hydrophobic part (hairpin) submerged into the oleo
some matrix (TAGs), all oleosins contain socalled
“proline knot”. It is supposed that this proline knot is
responsible for oleosin targeting to oleosomes [89].
Some seed tissues accumulate TAGs, which cannot
be mobilized during germination. For example, this
occurs in the endosperm of some cereals, which cells are
subjected to apoptosis during maturation [90] and thus
stored TAGs cannot be subjected to βoxidation during
seed germination. It is surprising that, in the endosperm
of mature oat seeds, large oily drops are observed, which
can be related to a deficit of oleosins [34].
According to the most common model of oily
inclusion biogenesis, proteins involved in the metabo
lism of neutral lipids are concentrated in the well
defined regions of ER, and enzymes involved in the
TAG biosynthesis are found among these proteins
[31]. TAGs are synthesized in the ER and accumulate
between two membrane layers of the ER [91]. Since
newly formed neutral lipids cannot insert into the
membrane, they accumulate in the hydrophobic
No. 1 2014
10 SIDOROV, TSYDENDAMBAEV
region between the two membrane layers, producing
oil drop (Fig. 4). During the continuous synthesis of
TAG, this oil drop continues to grow, and in its place a
“bud” is formed, which reaches a certain size, sepa
rates from the ER membrane, and turns out to be in
the cytosol as a separate compartment – oleosome
[30, 91].
At the same time, as it was mentioned above, there
are some plants (oil palm, avocado, olive, sea buck
thorn, and others), in which a significant amount of
oil is contained not only in the seeds, but also in the
juicy outside seed fruit parts, TAGs of which are not
used by the plants themselves as a source of organic
matter and energy. Such plants are called as plants
with oily mesocarp (POM).
POM are characterized by some specific features.
In particular, it was shown that oleosomes of their
endosperm are spherical, 0.5–2.0 μm in diameter,
whereas in the fruit parts outside the seeds (mesocarp,
hypantium, etc.) they are large, irregular in shape, 10–
25 μm in diameter. In addition, oleosomes in these
fruit parts differ sharply in the composition and con
tent of proteins in their membranes and in the compo
sition of TAG stored in them. In particular, oleosomes
of the endosperm and embryo, as a rule, contain much
protein (up to 10% of their weight), whereas the con
tent of protein in oleosomes in olive and avocado
mesocarp contain insignificant amounts of protein
[92]. With regard to TAGs, the outside seed fruit parts
(mesocarp, hypanthium, arillus, etc.) of many POM
fruit contains highly saturated oil with predominance
of C16 FAs, palmitic and hexadecenoic (or С18:1 [4, 5]),
while the same fruit seeds accumulate TAGs rich in
polyunsaturated С18 FAs, linoleic and αlinolenic
[7, 93].
This feature of POM plants was studied on the
example of the sea buckthorn hypanthium by analyz
ing the changes of positionspecies composition of its
TAGs during fruit ripening [93]. The obtained experi
mental data permitted the authors to suggest the hypo
thetic pathway of TAG biosynthesis (Fig. 5) contain
ing unsaturated FAs in the sn2 position of the mole
cule, which is different from the Kennedy pathway
[35]. The authors suggest that this process includes at
least two stages. At the first stage, the pool 1 of free sat
urated and unsaturated FAs statistically esterifies rac1
OH groups of so far unknown C3 precursors, thus
forming the common metabolic pool of TAG precur
sors (pool 2) comprising five different rac1 compo
nents. At the second stage, the components of pool 2
are subjected to selected sn2 acylation at expense of
the pool of unsaturated FAs (pool 3); this process is
catalyzed by so far unknown “acyltransferase X”.
According to the suggested hypothesis, sn2 acylation
is a terminal but not intermediate stage of TAG mole
cule biosynthesis; it is controlled by the amount of
available FAs and the degree of affinity of separate FA
to esterification of sn2 groups of glycerol or its deriv
atives. In addition, from the results of the performed
RUSSIAN JOURNAL OF PLANT PHYSIOLOGY
analysis of positionspecies TAG composition during
fruit ripening, it follows that in the sea buckthorn
hypantium a direct Δ12desaturation of the oleic acid
residues occurs, which esterifies rac1 positions of
TAG molecule already after the formation of their
molecules [93], as this occurs in developing sunflower
seeds at temperature lowering [94].
PHYSIOLOGICAL CONTROL OF TAG
ACCUMULATION DURING SEED RIPENING
Energetic of FA and TAG Biosynthesis
FA synthesis is a process requiring a great amount
of energy, and thus it depends on the stoichiometrical
contents in the plastid stroma of ATP, NADPH, and
NADH, which are consumed during each successive
lengthening of the growing acyl chain during FA bio
synthesis [21]. ATP is required for carboxylation of
acetylCoA in the reaction of its conversion into mal
onylCoA by acetylCoA carboxylase, whereas the
two reductases of the FAsynthetase complex, 3
ketoacylACP reductase and enoylACP reductase,
are required in NADPH, and NADH, respectively.
Plastids of the endosperm cells in oily seeds should
accumulate ATP and reducers or synthesize them. The
import into chloroplasts of reducers or ATP synthe
sized in mitochondria or cytosol requires the presence
of specific transporters, such as ATP/ADP exchange
translocases, which are so far poorly studied in seeds
[21].
A potential source of ATP and reducers in plastids
may be the functioning of glycolytic enzymes: glyc
eroaldehyde3phosphate dehydrogenase results in
the NADH formation, whereas phosphoglycerate
kinase and pyruvate kinase are involved in ATP syn
thesis. Similarly, successive conversion of pyruvate
into acetylCoA by the pyruvate dehydrogenase com
plex accompanies NADH synthesis. The intraplastid
conversion of malate into pyruvate and then into
acetylCoA is another potential source of reducers for
FA biosynthesis because plastid malate dehydrogenase
releases NADPH [95]. As a result, oxidation of sugar
phosphates in the plastid pentose phosphate cycle can
also lead to the formation of reducers, because two
dehydrogenases of this cycle, glucose6phosphate
dehydrogenase and gluconate6phosphate dehydro
genase, are involved in NADPH production [96].
Oxygen as a Factor Limiting Oil Accumulation in Seeds
Oxygen deficiency is evidently a characteristic fea
ture of plant tissues with a small surface area relative to
volume, for example, of specialized tissues of organs
accumulating storage compounds, oils in particular
[97]. This phenomenon is especially pronounced in
seeds with their low permeability for gases of maternal
tissues, in particular, their coats (nut shell, pod walls,
etc.). Thus, in rape seeds grown in the light in the pres
ence of 21% oxygen, the content of oxygen in the
Vol. 61 No. 1 2014
 BIOSYNTHESIS OF FATTY OILS IN HIGHER PLANTS 11

Oleosin
and other proteins

ER

Ribosomes Oleosomes of seeds, 0.5–2.0 μm

TAGs

ER

Ribosomes
Oleosome of fruit parts
outside the seeds

Fig. 4. Oleosomes of seeds and fruit parts outside the seeds.

inner pod space was reduced to 17% and in seeds, to
0.8% [97]. Such low oxygen content may be related to
a decrease in the ATP/ADP ratio and subsequent lim
itation of biosynthetic processes in ripening seeds.
Correspondingly, in the seeds of plants grown at a low
ered oxygen content, the rate of lipid production
reduced, whereas an increased oxygen content
resulted in the enhanced oil accumulation [97].
Despite the low oxygen content and reduced respira
tion rate, the plant regulates strictly seed metabolism
to prevent fermentation and to avoid hypoxia and
anoxia. Switching cell energy metabolism on the path
ways requiring less ATP and more efficiently using
oxygen is an effective response to conditions of low
oxygen in a medium. This may be achieved through
the activation of the metabolic pathways associated
with the metabolism of inorganic pyrophosphate.
Thus, in ripening arabidopsis seeds, pyrophosphate
dependent phosphofructokinase was activated and the
activity of ATPdependent phosphofructokinase was
declined [98]. By processing at the low oxygen con
centration “unnecessary” energy reserves in impor
tant metabolic processes, pyrophosphatedependent
metabolic pathways can prevent anoxia in the devel
oping embryo. However, the significance of the pyro
phosphatedependent metabolism in oilseeds remains
in doubt [99].
The limitation of the lipid metabolism intensity is
also the part of the adaptive response to the lowered
oxygen concentration [97]. Taking into account the
clear role of oxygen availability in the limitation of
TAG accumulation under definite growth conditions,
it may be concluded that this factor has little effect on
the intracellular starch accumulation. Experiments
with developing seeds of Brassicaceae plants allow a
postulation that oxygen plays a leading role in the bal
ance between storage lipids and starch [99]. Moreover,
under natural conditions, starch was positioned in the
maize grains in the oxygenpoor endosperm regions,
whereas TAGs, in contrast, accumulated in the
peripheral regions relatively enriched in oxygen, in the
embryo, aleurone layer, and in the layer of vitrious
endosperm [100].
Effect of Light and Photosynthesis in Embryo Tissues
on Storage Compound Metabolism
While plastids isolated from maturing castor bean,
safflower, and sunflower seeds lack chlorophyll, some
oily seeds, such as soybean, flax, and rape seeds con
tain chloroplasts and use photosynthetic processes as
an energy source in the biosynthesis of lipids and oil
accumulation. In this case, since the permeability of
the tissues surrounding the seed (e.g., pods) for light is
very low [100], the pigment composition of green
embryos exhibits traits of adaptation to low light, in
particular, the ratio of chlorophyll a/b therein is lower
than in leaves [101].
Although some seeds contain chloroplasts with the
thylakoid structure and a set of enzymes typical for
photosynthetically active plastids, they are storage
organs and use mixed biosynthetic pathways
(autotrophic and heterotrophic) for maintaining ana
bolic reactions [96]. The behavior of chloroplasts in
these seeds are often called as photoheterotrophism. It
was shown that the conditions of illumination affected

RUSSIAN JOURNAL OF PLANT PHYSIOLOGY Vol. 61 No. 1 2014

12 SIDOROV, TSYDENDAMBAEV

P+H+O+L
Total FAs
(Pool 1)

Sn1,3 RD of pool 1 FAs First stage

among unknown C3 of TAG biosynthesis
precursor molecules

PH + PP + HH + PO + HO
Putative diacyl
P+O+L
rac1 components
Unsaturated FAs
(Pool 2)
(Pool 3)
Second stage of TAG biosynthesis

Sn2 RD of pool 3 FAs among pool 2

components by putative

RD
“acyltransferase X” 1
6:
1
fС

of
o

С1
RD

8:
2
TAGs
PHH, PHP, HHH PHO, HHO RD of С18:1 of Group III
(see Table 2 in [93])
Group I TAGs
TAGs of Group II
(see Table 2 in [93])

Partial cisΔ12desaturation of
rac1O residues in TAGs

PHL, HHL
TAGs of Group IV

Fig. 5. Hypothetical pathway of TAG biosynthesis in the hypanthium of sea buckthorn fruit [93].
H, L, O, and P – residues of hexadecenoic, linoleic, oleic, and palmitic acid, respectively. RD—random distribution.

strongly oil accumulation in soybean, rapeseed, and
arabidopsis seeds: under high light, oil content in seeds
was higher than in the control group [101, 102]. Light
affects indirectly oil accumulation in seeds, influenc
ing diverse metabolic processes.
Primarily in the lightdependent reactions ATP
and NADP are produced, which can be used by the
cell for the FA synthesis in plastids [103]. In green
seeds, like in the leaves, the balance between cyclic
and noncyclic photophosphorylation can regulate the
synthesis of ATP and NADP [104]. It is interestingly
that a possibility to use light by oily seeds is reversely
dependent on the activity of the tricarboxylic acid
cycle, which also provide them with ATP. In sun
flower embryos devoid of chlorophyll, 100% of ATP
consumed for TAG biosynthesis is produced during
mitochondrial respiration [105], whereas in the green
rapeseed seeds the Krebs cycle is inactive and mito
chondria produce not more than 22% of ATP spent for
FA and TAG biosyntheses [106].
In addition, the oxygen produced in photosynthe
sis, supporting the synthesis of ATP and preventing
anoxia, in a local scale can play a key role in keeping
the rate of the lipid biosynthesis [100]. The sites of
oxygen photosynthetic production and those of its
consumption may differ. Thus, in maturing soybean
seeds, photosynthetic processes are localized in the
abaxial part of the seed (the external parts of cotyle
dons), whereas TAGs are synthesized in its adaxial
(inner) part. It is shown that oxygen produced during
photosynthesis diffuses into the adaxial part of the
embryo and enhances TAG synthesis there [107].
Finally, activation of the Rubisco “bypass”
improves carbon fixation during oil synthesis, com

RUSSIAN JOURNAL OF PLANT PHYSIOLOGY Vol. 61 No. 1 2014

 BIOSYNTHESIS OF FATTY OILS IN HIGHER PLANTS
pensating for the loss of CO2 in the reactions involving
pyruvate dehydrogenase [108]. Sunflower embryos
devoid of chlorophyll convert compounds – sources
of carbon, into storage TAGs at the efficiency not
exceeding 50%, whereas in green seeds more than 80%
of all carbon sources are metabolized into oil, as it was
shown for rapeseed [102, 105]. It is also established
that the oil biosynthesis in green seeds occurs most
actively in the daytime and drops sharply to the night
[101]. Likely that the reduction in the intensity of stor
age lipid biosynthesis in the dark is a strategy aimed at
reducing the losses of carbon in the form of CO2.
Thus, by activation of storage TAG biosynthesis in the
daytime, plants can efficiently increase the rate of oil
accumulation in the seeds [102].
Effect of Temperature Conditions on Oil Biosynthesis
in Seeds
Among environmental factors affecting oil accu
mulation in seeds, temperature is a primary factor for
some plant species, which usually plays an important
role in the control of oil FA composition [109]. Like
membrane lipids, storage lipids (TAGs) of plants
grown at lowered temperatures are characterized by
the increased content of PUFA, i.e., the unsaturation
index of TAGs is reversibly correlated with tempera
ture [110]. However, this dependence is differently
pronounced in different plant species. Thus, the
degree of safflower oil unsaturation only weakly
depends on temperature [111], in soybean and rapeseed
seeds – moderately [110, 112], and in sunflower –
strongly [113].
Several mechanisms explaining the effect of tem
perature on the FA composition of TAG stored in the
seeds are suggested. Primarily, plant response to
change in the temperature is related to Δ12desaturase
of FA encoded by the FAD2 gene, which activity is reg
ulated by transcriptional and posttranscriptional fac
tors [114]. Among posttranscriptional mechanisms
controlling FAD2 gene activity, longterm effect of
temperature is determined by the incapability of seed
specific isoforms of Δ12desaturase to function nor
mally under low ambient temperatures [115], whereas
shortterm effect is related to the temperature influ
ence on the content of available oxygen in seeds,
which determines the level of oxygencontaining co
substrates of Δ12desaturase [116]. The high thermo
stability and low dependence on the available oxygen
of the enzyme encoded by the FAD2 gene in safflower,
as compared with the corresponding enzyme of sun
flower, can explain why FAcomposition of safflower
TAGs, as distinct from sunflower TAGs, is essentially
independent of ambient temperature [117]. A temper
ature dependence of Δ9stearoylACP desaturase
gene expression was also demonstrated, and this can
also affect TAG FAcomposition [118].
RUSSIAN JOURNAL OF PLANT PHYSIOLOGY Vol. 61
13
Unusual Plant Optically Active
3Acetyl1,2diacylsnglycerols
As it was noted above, along with TAGs of a com
mon composition some plants can accumulate
unusual optically active NAGs, one or two positions of
which molecules may be acylated not by usual FA res
idues but by the residues of phenylalkanoic, phenyl
alkenoic, phenylcarbonic, or lowmolecularweight
aliphatic acids, formic and acetic ones.
The residue of acetic acid in storage TAGs of plant
seeds was identified for the first time still in 1948:
Kaufman and Keller found them in the seeds of Impa
tiens roylei (Balsaminaceae) [119]. These authors
studied the FAcomposition of TAGs and suggested
that the predominant molecular species of this NAG
type, acetylated DAGs (acDAGs), is 3acetyl1,2
diparinarinsnglycerol. Similar data confirming the
high content of acDAGs with unusual αparinarinic
acid (Δ9,11,13,1518:4) were obtained later for one
more species of impatiens (I. edgeworthii) [12]. Spe
cific features of other impatiens species (I. pallida and
I. capensis) fruits and the oil and αparinaric acid
accumulation in them during ripening were studied
much later; however, the FAcomposition of acDAGs
containing αparinaric acid was presented only for
mature seeds [120].
acDAGs containing the residues of usual FAs in
sn1,2 positions were described by Kleiman et al. in
1967 in the seed oils of 13 plant species belonging to
unrelated families Celastraceae, Lardizabalaceae,
Ranunculaceae, and Rosaceae [11]. It was shown that
seed oil of two species of Celastrus and Akebia, four
species of Euonymus, one species of Gymnosporia, and
one species of Maytenus, belonging to the Celas
traceae family contained both NAG classes, TAGs and
acDAGs, and the proportion of the latter was from 68
to 98% of total storage glycerolipids, whereas the con
tent of TAGs did not exceed 2–24%. In three mem
bers of Ranunculaceae and Rosaceae, tested by Klei
man et al., the proportion of acDAGs in seed oil did
not exceed 32% [11]. However, the authors did not
present data about the content of these compounds in
seeds (as calculated per dry and fresh weight), their
FAcomposition, specific features of their position
species acDAG composition, and the composition of
other neutral lipids.
Specific features of FAcomposition and position
species composition of plant acDAGs were studied in
three species of Maytenus (Celastraceae) by GLCMS
(CI) [121]. The authors showed that seed oils of spe
cies tested (Maytenus ilicifolia, M. dasyclada, and
M. cassineformis) contained up to 99% of acDAGs and
not more than 1% of TAGs of the common composi
tion, which differ substantially from data presented by
Kleiman et al. [11]. Spitzer and Aichholz [121]
showed that among acDAGs of seed oil of plant spe
cies studied, 7–8 molecular species predominated:
their sum attained up to 76% of total acDAGs.
No. 1 2014
14 SIDOROV, TSYDENDAMBAEV
In 2010 Durrett et al. [51] showed that in winged
spindle (E. alatus) storage NAGs accumulated not
only in the seed endosperm but also in tissues of
embryo and arils. In this case, mature seed oil con
tained 94.5% of acDAGs, 1.6% of TAGs, and 1.3% of
1,2DAGs, whereas in arils there was 92.2% of TAGs,
0.5% of acDAGs, and 2.0% of 1,2DAGs. In addition,
it was shown that acDAGs were synthesized with
the involvement of the specific membranebound
Oacetyl acyltransferase DGAT1 manifesting a speci
ficity to both the acyls of usual FAs and the residue of
acetate, whereas DGAT2 from the same material did
not manifest such specificity. At the same time, the
authors did not present any data about similarity or
difference in the FAcomposition of TAGs and
acDAGs in the seeds and arils and also about oil con
tents in seeds and arils [51].
Thus, it can be concluded that, despite the rela
tively detailed study of the mechanism of plant TAG
biosynthesis, information on how the formation of oils
stored in different parts of the POM plant fruits, which
differ sharply from each other in the qualitative com
position and the nature of the enzyme systems provid
ing these pathways, are so far absent in the literature.
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Translated by N. Klyachko
Vol. 61 No. 1 2014