Phytochem Rev

DOI 10.1007/s11101-016-9463-3

Occurrence, metabolism, transport and function

of seven-carbon sugars
A. Keith Cowan

Received: 18 January 2016 / Accepted: 25 March 2016

Ó Springer Science+Business Media Dordrecht 2016

Abstract Seven-carbon (7-C) sugars and sugar of the corresponding phloem translocated heptuloses
alcohols are common in higher plants, algae, fungi following import into non-photosynthetic tissue. It is
and bacteria. The biochemical origin and physiolog- proposed that the major physiological function of 7-C
ical function of these monosaccharides in plants and sugars and heptitols, in addition to serving as a carbon
algae however is not well understood and has not been sink, involves metal ion chelation, translocation and
fully investigated. Here the occurrence, metabolism, remobilisation to fulfil nutrient requirements essential
and transport of heptuloses, heptitols, and heptoses are for growth and development.
integrated in accordance with function to emphasise
the importance of these apparently neglected sugars. Keywords Heptitols  Heptuloses 
This therefore is the first comprehensive synthesis of Mannoheptulose  Metabolism  Sedoheptulose
knowledge about 7-C sugar biochemistry, a relatively
underexplored area of carbohydrate biology that needs
Abbreviations
to be integrated into mainstream sugar research.
7-C Seven-carbon
Available information on the metabolism of heptu-
ADP-L,D-Hep ADP-b-L-Glycero-D-manno-heptose
loses, heptitols, and heptoses in Medicago sativa
CAM Crassulacean acid metabolism
(alfalfa), Persea americana (avocado), Primula sp.,
FBA Fructose-bis-phosphate aldolase
Kalanchoë pinnata, and the red alga Porphyridium sp.
FK Fructokinase
was thoroughly investigated and evaluated. Results
gro-manHep D-Glycero-D-manno-heptose
indicate that 7-C sugars share a common precursor and
Hep Heptose
are products of a TKT-dependent heptulose shunt in
HK Hexokinase
which Suc-derived Fru 6-P is converted either to Sed
INV Invertase
7-P or mannoheptulose 7-P or both in competent
Lgro-manHep L-Glycero-D-manno-heptose
tissues and species. In plants, free heptuloses probably
Moco Molybdenum cofactor
arise as a consequence of phosphatase activity,
PPP Pentose phosphate pathway
whereas heptoses appear to be formed by isomerisation
PCR Photosynthetic carbon reduction
Pi Inorganic phosphate
A. K. Cowan (&) SBPase Sed 1,7-bis-phosphatase
Institute for Environmental Biotechnology, Rhodes SPS Sucrose phosphate synthase
University (EBRU), Rhodes University, SPP Sucrose phosphate phosphatase
P.O. Box 94, Grahamstown 6140, South Africa SuSy Sucrose synthase
e-mail: a.cowan@ru.ac.za

123
 Phytochem Rev

TAL Transaldolase structures (Koch 2004). Other carbohydrates trans-

TKT Transketolase ported in vascular plants include polyols like sorbitol
TPP Thiamine diphosphate and mannitol (Noiraud et al. 2001), oligosaccharides
of the raffinose family (Rennie and Turgeon 2009),
and several seven-carbon (7-C) sugars and related
heptitols (Liu et al. 1999a, b).
Introduction
Amongst the best studied of the 7-C sugars are
mannoheptulose ((3S,4S,5R,6R)-1,3,4,5,6,7-hexahy-
Carbohydrates are synthesized by all photosynthetic
drox-2-heptanone; D-glycero-D-manno-heptulose), its
organisms and represent the chemical store of solar
corresponding sugar alcohol perseitol [(2R,3R,5R,6S)-
energy that is used to support life. More commonly
heptane-1,2,3,4,5,6,7-heptol; D-glycero-D-galacto-hep-
called sugars, these polyhydroxylated aldehydes and
titol), sedoheptulose (Sed; D-altro-2-heptulose) and the
ketones are classed as either simple (i.e. monosaccha-
related heptitol volemitol (D-glycero-D-manno-heptitol,
rides) or complex (i.e. disaccharides, oligosaccha-
a-sedoheptitol)]. The structures of these sugars are
rides, and polysaccharides). Both can fulfil structural
shown in Fig. 1.
(Peshev and Van den Ende 2013), signalling (Smee-
Information on 7-C sugars while sparse is of
kens et al. 2010; O’Hara et al. 2013; Yadav 2014),
considerable interest for several reasons. First, despite
transport (Doidy et al. 2012; Lemoine et al. 2013), and
a substantial literature very little is known about the
storage functions (Bihmidine et al. 2013). Together
occurrence, metabolism, transport and physiology of
with the disaccharide sucrose (Suc), the monosaccha-
7-C sugars (Cowan 2004); second, many studies on
rides glucose (Glc; D-hexose) and fructose (Fru; D-
mannoheptulose in human and animal systems indi-
hexulose) that cannot be further converted by hydrol-
cate that a major pharmacological function of this 7-C
ysis to more simple sugars are the most widely
sugar is inhibition of hexokinase (HK; EC 2.7.1.1) and
recognized and amongst the best studied. Simple
impaired Glc metabolism (Tullson and Aprille 1986;
sugars have the general formula CnH2nOn, where n is
Board et al. 1995; Scruel et al. 1998; Courtois et al.
at least three and rarely more than six although seven-,
2000); and third, exogenous mannoheptulose miti-
eight- and nine-carbon sugars have been identified
gates a variety of HK-related effects in plants. These
(Begbie and Richtmyer 1966). The acyclic forms are
include: attenuation of the effects of HK overexpres-
named according to three characteristics: the number
sion in tomato (Dai et al. 1999); mitigation of HK-
of carbon atoms; the carbonyl group; and, whether in
dependent signalling in D-allose-induced inhibition of
the D- or L-configuration (Table 1).
gibberellin-dependent leaf sheath elongation and seed
Monosaccharides play a central role in primary
a-amylase activity in rice (Fukumoto et al. 2011,
metabolism and link energy derivation by photosyn-
2013); and, effecting a rise in endogenous Suc and
thesis to cellular anabolic and catabolic needs, and are
restricting Glc-induced accumulation of trehalose-6-
the substrates for Suc biosynthesis. Typically, chloro-
phosphate and Suc in Arabidopsis thaliana seedlings
plast triose phosphates are exchanged for inorganic
(Yadav 2014). Together, this experimental evidence
phosphate (Pi) from the cytoplasm and converted to
suggests a role for mannoheptulose and possibly other
hexose phosphate via gluconeogenesis for Suc syn-
7-C sugars in the regulation of carbon flux.
thesis. Sucrose, the principal product of photosynthe-
As far as can be ascertained, there exists no
sis, is also the major transportable sugar in most
comprehensive overview of the biology of 7-C sugars
vascular plants. It is cleaved either by invertase (INV;
and the extensive literature, spanning more than a
b-D-fructofuranosidase, EC 3.2.1.26) or Suc synthase
century, has not previously been analyzed and assem-
(SuSy; UDP-D-glucose:D-fructose-2-a-glucosyltrans-
bled to provide a thorough commentary on this
ferase, EC 2.4.1.13) following entry into non-1photo-
important topic. Here, available information on the
synthetic sink tissues such as roots and reproductive
occurrence, metabolism, transport and function of 7-C
sugars and sugar alcohols is synthesized in an effort to
1 focus attention on these apparently neglected carbo-
Monosaccharide codes used are as defined by the Kyoto
Encyclopedia for Genes and Genomes (The KEGG Database hydrates and, as a first step toward elaborating their
2016). role in plants and algae.

123
Phytochem Rev
Table 1 Carbohydrate nomenclature showing the functional groups and the assignment of stereochemical configuration
No. of
Aldehyde Ketone Carbonyl group
Carbons
3 Triose Triulose
4 Tetrose Tetrulose
5 Pentose Pentulose
6 Hexose Hexulose
7 Heptose Heptulose
8 Octose Octulose
9 Nonose Nonulose
Fig. 1 Structural formulae of mannoheptulose, sedoheptulose,
and the related acyclic heptitols, perseitol and volemitol
Occurrence of seven-carbon sugars and sugar
alcohols
Mannoheptulose from avocado fruit was the first 7-C
sugar to be isolated and identified (LaForge 1916) and
its presence was subsequently confirmed in a range of
plants including gymnosperms from the family Tax-
odiaceae, numerous dicotyledonous families (includ-
ing commercially important families such as
Asteraceae, Lauraceae, Leguminosae, Moraceae,
Rosaceae, Rutaceae and Salicaceae) and the mono-
cotyledonous family Iridaceae (Okuda and Mori 1974;
Shaw et al. 1980; Quintanilla-Licea et al. 2012). The
heptahydroxy alcohol perseitol, also isolated from
avocado, was first described by Avequin in 1831 and
although a complex of perseitol and K? ions was
isolated and characterized from Scurrula fusca (Lo-
ranthaceae) leaves (Ishizu et al. 2001), very little is
known about the metabolism and physiology of this
non-structural carbohydrate. Perseitol is widely dis-
tributed in tissues and organs of Persea americana
(avocado) and is also present in avocado nectar (Liu
D/L Configuration
et al. 1995; Ish Am and Eisikowitch 1998). However,
perseitol concentration in honey from avocado is only
0.48 % of the total sugar content which is considered a
consequence of the relatively high amounts of man-
noheptulose (Dvash et al. 2002). Thus, aside from a
postulated role as a storage and transport sugar (Liu
et al. 1999a, b) there is very little published informa-
tion on the biosynthesis and breakdown of this
heptitol.
Other 7-C sugars and sugar alcohols that have
attracted attention include Sed and coriose (D-altro-3-
heptulose) isolated from leaves of Coriaria japonica
(Okuda and Mori 1974), volemitol (D-glycero-D-
manno-heptitol) and its epimer b-sedoheptitol (Kre-
mer 1978; Häfliger et al. 1999) also present in
avocado, and laminitol (D-4-C-methyl-myo-inositol)
and its epimer mytilitol (C-methyl-scyllo-inositol)
isolated from several algae belonging to the Rhodo-
phyceae, Phaeophyceae, and Chlorophyceae (Wöber
and Hoffmann-Ostenhof 1970a; Young 1972).
Sedoheptulose occurs ubiquitously in all photosyn-
thetic organisms. This 7-C heptoketose was first
isolated from aqueous extracts of Sedum spectabile
(LaForge and Hudson 1917). Later, Sed was extracted
and identified in the tropical fruits mango (Mangifera
indica) and papaya (Carica papaya) (Ogata et al.
1972) and quantified in amounts between 1.4 and
24.6 mg g-1 dry weight as a major component of
carrot (Daucus carota) root carbohydrates (Soria et al.
2009). Although free Sed is abundant in members of
the Crassulaceae such as Hylotelephium spectabile
(formerly Sedum spectabile) and Kalanchoë pinnata
(Ceusters et al. 2013), its presence in plant tissues from
a range of other families including Apiaceae, Aquifo-
liaceae, Euphorbiaceae, Lamiaceae, Primulaceae, and
Saxifragaceae suggests a wider occurrence (Tolbert
et al. 1957; Okuda and Mori 1974; Häfliger et al. 1999;
123

Soria et al. 2009). The monophosphate derivative of
Sed, Sed 7-P, is a key intermediate in both photosyn-
thetic carbon reduction (PCR) and the pentose phos-
phate pathway (PPP) and is proposed to function in
carbon partitioning and storage (Häfliger et al. 1999;
Ceusters et al. 2013). Volemitol is the predominant
phloem-mobile sugar in Primula (Primula 9 polyan-
tha) where it accounts for up to 24 % (mol mol-1) of
phloem sap carbohydrate (Häfliger et al. 1999). This
heptitol was initially considered as being confined to
certain sections of the genus Primula (Häfliger et al.
1999). However, it’s presence in the brown alga
Pelvetia canaliculata (Young 1972; Pfetzing et al.
2000) and the lichen Parmotrema cetratum (da Silva
et al. 1993) would appear to suggest that it too occurs
more widely in nature. Laminitol was first isolated
from Laminaria cloustoni and later from several
species of red and brown algae and the chlorophyte,
Chlorella fusca (Wöber and Hoffmann-Ostenhof
1970a). Again, and as with other 7-C sugars very
little is known about the metabolism and physiology of
either laminitol or mytilitol. There is evidence to
suggest that volemitol arises as an early product in
photosynthetic carbon reduction (Häfliger et al. 1999).
In contrast, laminitol appears to arise from phospho-
rylated derivatives of L-arabinose (L-Ara) via L-gluco-
heptulose (Wöber and Hoffmann-Ostenhof 1970b)
and may also function as an osmolyte (Ye et al. 2013).
Several fungi and yeasts, in addition to vascular
plants and algae, contain and appear to metabolize a
variety of heptoses, heptuloses and heptitols. For
example, the isomerization of D-glycero-D-galacto-
heptose to D-manno-heptulose by several yeast strains
has been demonstrated (Stankovič et al. 1986).
D-Glycero-D-manno-heptose has been detected in
Saccharomyces cerevisiae and Torulopsis versatilis
(Onishi and Perry 1972) and the heptitols, meso-
glycero-ido-heptitol and D-glycero-D-ido-heptitol,
have been reported as metabolic products of the
osmophilic yeast Pichia miso (Onishi and Perry 1965).
A comprehensive list of the various naturally
occurring heptoses, heptuloses, and heptitols isolated
and identified from plants, algae, and fungi is
presented in Table 2. Unfortunately, quantification
data are lacking. Identification in most instances was
by a combination of derivatization, paper and gas–
liquid chromatography, melting point determination,
crystallization, and infrared spectroscopy (Begbie and
Richtmyer 1966). As a consequence, there are little
123
Phytochem Rev
detailed mass spectrometry and/or nuclear magnetic
resonance spectroscopic data available for the natu-
rally occurring 7-C sugars isolated thus far. By
comparison, detailed crystal and molecular structures
have been established for the acyclic heptitols per-
seitol, volemitol and b-sedoheptitol (Angyal and Le
Fur 1984; Köll et al. 1991). Thus, perseitol is the only
heptitol that assumes a planar zigzag conformation,
whereas volemitol has a bent or sickle conformation
with C1–C6 in the planar zigzag conformation and the
C7 orientated parallel to the oxygen at C4.
On the origin of heptoses and heptuloses
As already mentioned, there is limited information
available on the metabolism of 7-C sugars in plants
and algae, and the biosynthetic origin and catabolism
of these monosaccharides has remained largely unex-
plored. This contrasts with the glycero-manno-heptose
component of the oligosaccharide in lipopolysaccha-
rides from gram-positive and negative bacteria, in
which the pathways for the biosynthesis of the
nucleotide-activated glycero-manno-heptoses have
been elucidated (Valvano et al. 2002; Morrison et al.
2005; McArthur et al. 2005). In brief, the ADP-b-L-
glycero-D-manno-heptose (ADP-L,D-Hep) biosyn-
thetic pathway comprises five steps, involves four
enzymes, and starts with isomerization of Sed 7-P. An
isomerase, GmhA, generates D,D-Hep 7-P which is
then phosphorylated by the kinase activity of the bi-
functional enzyme, HldE, to produce b-D,D-Hep 1,7-
bisP. A phosphatase, GmhB, cleaves the phosphate at
C-7 to give b-D,D-Hep 1-P, and the second activity of
HldE catalyses a pyrophosphorylation reaction to give
ADP-D,D-Hep. Finally, ADP-L,D-Hep is generated
by a reversible epimerization catalyzed by the enzyme
ADP-b-L-glycero-D-manno-heptose 6-epimerase (HldD
or AGME).
Several heptoses have been extracted and unequiv-
ocally characterized from plant tissues. D-Glycero-D-
galacto-heptose was identified from avocado fruit,
while D-glycero-D-gluco-heptose and its b-anomer, D-
glycero-D-manno-heptose, were isolated from roots of
Primula offininalis (Sephton and Richtmyer 1963;
Begbie and Richtmyer 1966) and the structures of
these sugars are shown in Fig. 2. Although no
information on the biosynthetic origin of these hep-
toses is available, it is noteworthy that certain yeast
Phytochem Rev

Table 2 Distribution of naturally occurring heptoses, heptuloses and heptitols in plants, algae, and fungi (see text for references)
Species Plant part 7-C Sugar/sugar alcohol

Angiosperms
Persea americana Fruit D-glycero-D-galacto-heptose, D-allo-heptulose; D-altro-2-heptulose;
D-talo-heptulose; D-D-glycero-D-manno-heptulose; D-glycero-
D-galacto-heptitol; D-glycero-D-manno-heptitol

Leaves D-altro-2-heptulose

Seed D-manno-heptulose; D-glycero-D-galacto-heptitol; D-glycero-

D-manno-heptitol

Primula officinalis Roots D-glycero-D-manno-heptose; D-glycero-D-gluco-heptose;

D-allo-heptulose; D-altro-2-heptulose; D-altro-3-heptulose;
D-glycero-D-gluco-heptitol; D-glycero-D-manno-heptitol

Primula elatior Roots D-altro-2-heptulose; D-glycero-D-manno-heptulose;

D-glycero-D-manno-heptitol

Mangifera indica Fruit D-altro-2-heptulose

Carica papaya Fruit D-altro-2-heptulose

Daucus carota Roots D-altro-2-heptulose

Coriaria japonica Leaves D-altro-2-heptulose; D-altro-3-heptulose;

Papaver somniferum Capsules D-altro-2-heptulose; D-glycero-D-manno-heptulose

Ficus carica Fruit D-altro-2-heptulose; D-glycero-D-manno-heptulose

Leaves D-altro-2-heptulose; D-glycero-D-manno-heptulose

Medicago sativa Leaves D-altro-2-heptulose; D-glycero-D-manno-heptulose

Cannabis sativa Leaves and stems D-altro-2-heptulose; D-glycero-D-manno-heptulose

Vitis vinifera Wine D-altro-2-heptulose; D-glycero-D-manno-heptulose

Fabiana imbricata Dried herbage D-glycero-D-manno-heptulose; D-glycero-D-galacto-heptitol

Hylotelephium spectabile Leaves and stems D-altro-2-heptulose; D-glycero-D-manno-heptitol;

(formerly Sedum spectabile) D-glycero-D-gluco-heptitol; D-glycero-D-galacto-heptitol

Green algae
Cladophora sericea – D-glycero-D-manno-heptulose; D-glycero-D-manno-heptitol

Chlorella fusca – D-4-methyl-myo-inositol; C-methyl-scyllo-inositol

Brown algae
Laminaria cloustoni – D-4-methyl-myo-inositol

Macrocystis pyrifera – D-4-methyl-myo-inositol

Desmarestia aculeata – D-altro-2-heptulose

Pelvetia canaliculata – D-altro-2-heptulose; D-glycero-D-manno-heptitol

Red algae
Porphyridium sp. – D-4-methyl-myo-inositol; L-gluco-heptulose; 1-deoxy-L-gluco-heptulose
Porphyra sp. – D-glycero-D-manno-heptitol

Fungi
Parmotrema cetratum Lichen D-glycero-D-manno-heptitol

Lactarius volemus Stipe and cap D-glycero-D-manno-heptitol

Yeast
Saccharomyces cerevisiae – D-glycero-D-manno-heptose

Torulopsis versatilis – D-glycero-D-manno-heptose

Pichia miso – D-glycero-D-ido-heptitol, meso-glycero-ido-heptitol

123

Fig. 2 Fischer projection formulae of the major heptoses,
heptuloses and heptitols isolated and identified in extracts from
vascular plants
strains isomerize D-glycero-D-galacto-heptose to man-
noheptulose and that the reverse reaction apparently
does not take place (Stankovič et al. 1986).
In contrast to heptose formation, a number of
studies have explored the biosynthesis of plant hep-
tuloses and in particular, the origin of mannoheptu-
lose, using exogenous substrates. These revealed that
when the hexulose L-sorbose (L-Sor) was fed to shoots
of Medicago sativa via the transpiration stream,
L-galacto-heptulose was formed (McComb and Ren-
dig 1961). Likewise, the administration of pentoses to
leaves of Medicago sativa revealed that D-ribose
(D-Rib) produced D-altro-2-heptulose, L-Ara gave
D-gluco-heptulose, D-xylose (D-Xyl) gave D-ido-hep-
tulose, and L-lyxose (L-Lyx) gave L-galacto-heptulose
(Rendig and McComb 1962a, b). Each of the four
administered pentoses induced accumulation of a
specific heptulose that has the C2 hydroxyl group in
the D configuration, and this is retained in the resultant
heptuloses which all show the D-threo configuration at
C3–C4. In contrast, enantiomers of these pentoses in
123
Phytochem Rev
which the C2 hydoxyl group is in the L configuration
did not induce heptulose accumulation (Rendig and
McComb 1962b). A detailed stereochemical analysis
using D- and L-tetroses confirmed that structural
integrity of the substrate was retained during conver-
sion to the resultant heptulose (Rendig and McComb
1964). Furthermore, hexuloses either with L-erythro,
D-erythro or L-threo configuration at C3–C4 did not
cause heptulose accumulation whereas L-Sor, which
has the D-threo configuration, induced accumulation
of L-galacto-heptulose. Based on these results, which
showed that tetroses and pentoses are converted with
steric specificity into the corresponding heptuloses, it
was suggested that incorporation of hexuloses
depended on a C3–C4 configuration characteristic of
transketolase (TKT; EC 2.2.1.1) substrates. Even so,
no information about the pathway of D-mannoheptu-
lose biosynthesis, which has the L-erythro configura-
tion at C3–C4 was obtained from these studies.
When D-Rib was fed to plant tissues accumulation
of D-Sed was observed (Rendig and McComb 1962a,
b). Even so, the biosynthetic origin of free Sed, which
has the L-threo configuration at C3–C4, has yet to be
unequivocally established, although more recent evi-
dence suggests that it arises by dephosphorylation of
the PCR cycle and PPP intermediate Sed 7-P (Ceusters
et al. 2013).
Sugars that have the D-threo configuration at C3-C4
similar to L-Sor include D-Fru, D-Glc and D-mannose
(D-Man). Exogenous application of the ketose D-Fru
did not induce heptulose accumulation in leaves of
Medicago sativa (Rendig and McComb 1964). This
was attributed to the rapid incorporation of D-Fru into
competing pathways, which was supported by the
observation that D-erythrose (D-Ery), a product of TKT
action on D-Fru, induced less heptulose accumulation
than either D- or L-threose (Tho). Transketolase
typically catalyzes the in vivo reversible transfer of
a-hydroxyacetyl from a ketose phosphate (donor) to
an aldose phosphate (acceptor). A TKT purified from
leaves of Spinacia oleracea (spinach) used D-Fru,
L-Sor and D-Sed in addition to hydroxypyruvate as
donors to form Sed when D-Rib was the acceptor while
gluco-heptulose was formed from D-Sed and L-Ara
(Villafranca and Axelrod 1971). Interestingly,
although the rate of heptulose formation was substan-
tially greater when phosphorylated derivatives were
used as substrates, it was not essential that the donor be
phosphorylated.
Phytochem Rev
Fig. 3 Scheme depicting TKT formation of L-Tho from L-Sor (shown in blue) and TAL-catalyzed formation of L-galacto-heptulose
with L-Tho as the acceptor of the dihydroxyacetone moiety from D-Fru (shown in red)
Together, these results confirmed findings from
feeding experiments using excised leaves of Medicago
sativa. Importantly, TKT-catalyzed formation of hep-
tuloses resulted in retention of the pentose chain from
C3 through C7, and required a hydroxyl group at C2 in
the D-configuration of the acceptor. Thus, it might be
expected that the product of TKT action on L-Sor
would be L-Tho. However, introduction of L-Sor to
plants gave L-galacto-heptulose (McComb and Ren-
dig 1961). Furthermore, feeding L-Tho to plants also
yielded L-galacto-heptulose (Rendig and McComb
1964). To explain this anomaly it was suggested that
L-Tho and a dihydroxyacetone moiety, derived from a
suitable donor, condensed and that by transaldolase
(TAL; EC 2.2.1.2) activity, L-galacto-heptulose was
formed. Thus, and as inferred by the authors, L-Tho
acts as an acceptor for the 3-C fragment from D-Fru in
the TAL-catalyzed formation of L-galacto-heptulose.
The scheme is illustrated in Fig. 3. Both D-Fru and
D-Sed have long been recognized as 3-C donors in
TAL-catalyzed reactions, and formation of D-Sed 7-P
from D-Fru 6-P has been confirmed for a number of
plant and fungal systems including Solanum lycoper-
sicon (Caillau and Quick 2005), Blastobotrys
adeninivorans (Fiki et al. 2009), Fusarium oxysporum
(Kourtoglou et al. 2008) and Saccharomyces cere-
visiae (Huang et al. 2008). Neither D- or L-Ery, nor D-
or L-Tho gave rise to mannoheptulose when these were
fed to leaves of Medicago sativa (Rendig and
McComb 1964), indicating that a TAL is unlikely to
be involved in the direct synthesis of this 7-C ketose.
Furthermore, the only aldopentose acceptor available
for mannoheptulose formation by TPP-dependent
TKT action is D-Ara. However, application of free
D-Ara to excised Medicago sativa leaves did not cause
accumulation of any heptulose. Even so, it is note-
worthy that avocado, a very rich source of manno-
heptulose, also contains substantial amounts of Ara
(Jeong and Huber 2004), and D-Ara is routinely used in
the commercial scale chemical synthesis of
mannoheptulose.
In addition to 7-C sugars, several 8- and 9-C
sugars have also been isolated and identified from
vascular plants (Charlson and Richtmyer 1960;
Sephton and Richtmyer 1966; Bianchi et al. 1991)
and by theoretical chemical analysis, grouped
according to probable biosynthetic origin of the C3-
C4 carbons. The complement of 7-, 8- and 9-C sugars
isolated and identified from avocado is listed in
Table 3. The first group includes sugars that have a
D-threo configuration at C3-C4 (i.e. the D-Xul con-
figuration at C1-C4), and these include Sed,
D-glycero-D-galacto-octulose, D-erythro-L-gluco-
nonulose, and D-erythro-L-galacto-nonulose. The
second group all has an L-erythro configuration at
C3-C4 and includes mannoheptulose, D-talo-heptu-
lose, and D-glycero-D-manno-octulose (Sephton and
Richtmyer 1966). Accordingly, this grouping of
avocado higher carbohydrates based on the diastere-
omeric configuration at C3-C4 would also appear to
123

Table 3 Higher carbon (7-C, 8-C, and 9-C) sugars isolated
and identified from avocado
No. of Sugar
carbons
7 D-glycero-D-galacto-heptose
D-talo-heptulose
D-altro-2-heptulose (Sedoheptulose)
D-glycero-D-manno-heptulose (Mannoheptulose)
D-glycero-D-galacto-heptitol (Perseitol)
D-glycero-D-gluco-heptitol (b-Sedoheptitol)
D-glycero-D-manno-heptitol (a-Sedoheptitol;
Volemitol)
8 D-glycero-D-manno-octulose
D-glycero-L-galacto-octulose
D-erythro-D-galacto-octitol
9 D-erythro-L-gluco-nonulose
D-erythro-L-galacto-nonulose
suggest that mannoheptulose and Sed
independently.
Seven-carbon sugar metabolism
Pathways of seven-carbon sugar metabolism
in vascular plants
Early studies on 7-C sugar metabolism revealed that in
avocado leaves, brief photosynthesis in a 14CO2 atmo-
sphere produced labeled mannoheptulose monophos-
phate as well as Sed monophosphate and intermediates
of carbohydrate synthesis (Nordal and Benson 1954).
Furthermore, when 14CO2 was supplied to avocado
leaves and leaf discs, Suc and mannoheptulose were
labeled rapidly in the light whereas perseitol was
labeled more slowly (Bean et al. 1962). Also, perseitol
continued to accumulate radiolabel in the subsequent
dark period, whereas 14C in mannoheptulose declined
suggesting that perseitol arises independently of pho-
tosynthesis. This contrasts with a more recent study in
which both mannoheptulose and perseitol appeared to
be labeled similarly following incubation of avocado
leaves in a 14CO2 environment (Liu et al. 2002).
However, calculation of the specific radioactivity
(Bq.lmol-1) associated with the individual sugars
reveals the following order of labeling: Suc [ manno-
heptulose [ perseitol, confirming earlier results (Bean
et al. 1962). Similar findings were obtained following
123
Phytochem Rev
analysis of avocado fruit after pulsed application of
[U-14C]-Suc via the pedicel. It was shown that while the
bulk of the 14C was in Glc (33 %) and Fru (41 %), no
14
C was detected in the perseitol fraction whereas
mannoheptulose contained 12 % of the total radioac-
tivity supplied (Cowan 2004). This result suggests that
Suc is not a precursor of perseitol in avocado fruit, and
that there is no apparent metabolic relationship between
the 7-C sugar mannoheptulose and the alditol perseitol.
Further studies using cell free extracts prepared
from mature source leaves of avocado supplied either
dihydroxyacetone phosphate ? Ery 4-P or Rib
5-P ? Xul 5-P, produced Sed 1,7-bis-P and manno-
heptulose and Sed 7-P, respectively (Liu et al. 2002).
Based on these results it was concluded: 1) that
mannoheptulose arose by an aldolase-catalyzed reac-
tion involving condensation of triose and tetrose
phosphates to form Sed 1,7-bisP followed by isomer-
ization; and, 2) that Sed 7-P was not metabolized
arise further to mannoheptulose. Given the steric specificity
with which tetrose phosphates are incorporated into
heptuloses it is unlikely that D-Ery would yield
mannoheptulose. Unfortunately, the configuration
(i.e. D- or L-) of the substrates used in this study is
unknown, and dihydroxyacetone phosphate was sup-
plied at \100 % (mol mol-1) of the co-substrate Ery
4-P and may therefore have been limiting.
The interconversion of mannoheptulose and per-
seitol has also been studied using crude enzyme
preparations of light grown and etiolated avocado
tissue (Tesfay et al. 2012). Activity was measured
indirectly by quantifying either NADP? or NADPH at
340 nm. It was revealed that the oxidase activity was
highest in cotyledons of etiolated seedlings whereas
the reductase activity prevailed in light grown
seedlings. This coupled with electrophoretic detection
of a putative aldoketose reductase/oxidase was taken
as evidence supporting mannoheptulose $ perseitol
interconversion in avocado seed and fruit tissues.
In another study, pulse-chase radiolabelling of
polyanthus leaf discs using 14CO2 indicated formation
of volemitol as a product of photosynthesis via Sed, in
a reaction catalyzed by an NADPH-dependent ketose
reductase with high substrate affinity (Häfliger et al.
1999). Activity of this ketose reductase, believed to be
Sed reductase, was apparently only detected in
volemitol-containing tissues. This conclusion was
based on recombination studies using enzyme
extracted from ‘‘volemitol-lacking avocado leaves’’.
Phytochem Rev
However, volemitol is present in avocado and has been
identified in extracts of both seed and mesocarp (fruit
flesh) tissue (Table 2) (Richtmyer 1970a). Photosyn-
thetic CO2 assimilation by avocado fruit is 0.4–2.5 %
that of fully expanded leaves (Whiley et al. 1991).
Thus, the bulk of volemitol in seed and fruit flesh tissue
is probably derived from sources other than the fruit.
In summary, results from studies using radiola-
belled precursors indicate that for mature leaves (i.e.
source tissue), photosynthetically fixed 14CO2 is a
precursor of Suc, mannoheptulose P, Sed P and
volemitol, and to a lesser extent, perseitol. In sink
tissues; radiolabel from Suc is incorporated, via
hexose/hexulose into mannoheptulose but apparently
not into perseitol.
Seven-carbon sugar metabolism in algae
In algae, evidence indicates that the mechanism of Suc
production is similar to that of the much more recently
evolved higher plants, and occurs via Suc-phosphate
synthase (SPS; UDP-glucose: D-fructose-6-phosphate
glucosyl-transferase, EC 2.4.1.14)/phosphatase (SPP,
sucrose-6-phosphate phosphohydrolase, EC 3.1.3.00),
by a complex termed SPS/SPP (Porchia and Salerno
1996). Furthermore, the products of algal photosyn-
thesis can be partitioned between storage polysaccha-
rides (e.g. green algae typically store starch; red algae
store glucans of linear a-1,4 and branched a-1,6
glycosidic linkages; and, brown algae store water-
soluble laminarin and chrysolaminarin composed of b-
1,3 linkages branched at the C-2 and C-6 of Glc) and
fixed carbon products such as mannitol, Ara, and
glycerol, which are either translocated or excreted
(Radakovits et al. 2010). Mannitol is a major compo-
nent of photosynthate translocated in kelps and is
widespread in the Phaeophyta and Rhodophyta, but
less so in the Chlorophyta (Lewis and Smith 1967).
Other sugar alcohols shown to be present in algae
include sorbitol, ribitol, erythritol, and the heptahy-
droxy alcohol, volemitol.
Large amounts of volemitol occur in Pelvetia
canaliculata, and mannoheptulose and Sed have been
found in species of brown, red and green algae. In the
brown alga Pelvetia canaliculata, which contains both
mannitol and volemitol, mannitol biosynthesis was
confirmed by the well-documented reduction of Fru 6-P
catalyzed by a NADH-linked dehydrogenase (Kremer
1977). In addition, a putative NADH-requiring enzyme
with high specificity for Sed 7-P was detected. Given
these findings it was proposed that formation of
volemitol in Pelvetia follows the pathway: Sed
7-P ? volemitol 1-P ? volemitol (Kremer 1977).
Based on the stereochemistry of laminitol and
knowledge of myo-inositol biosynthesis both L-gluco-
heptulose and its reduction product 1-deoxy-L-gluco-
heptulose were proposed to function as precursors of
this 7-C cyclitol in the red alga Porphyridium (Wöber
and Hoffmann-Ostenhof 1970b). Radioactive L-gluco-
heptulose was produced biosynthetically by incubat-
ing L-Ara fed leaves of M. sativa in a 14CO2
environment, and after isolation and purification,
supplied to cultures of Porphyridium sp. supple-
mented with 5 % CO2. Radioactivity from L-gluco-
heptulose was readily incorporated into laminitol and
by trapping, the presence of endogenous pools of both
L-gluco-heptulose and 1-deoxy-L-gluco-heptulose was
demonstrated. Furthermore, incorporation of radioac-
tivity from labeled L-Ara into laminitol indicated a role
for a TKT in the biosynthesis of this 7-C cyclitol. By
applying [U-14C]-laminitol to cultures of Chlorella
fusca, labelled mytilitol could be isolated suggesting
that this 7-C sugar arises by epimerization of
laminitol (Wöber and Hoffmann-Ostenhof 1970a).
Further detailed metabolic studies using Porphyridium
sp. revealed that the precursors most efficiently
converted to laminitol were D-mannoheptulose, L-
gluco-heptulose, 1-deoxy-D-mannoheptulose, and
1-deoxy-D-mannoheptulose (Woloszczuk and Hoff-
mann-Ostenhof 1974). Moreover, the intermediates
isolated were confirmed as phosphorylated, after
recovery from the acid fraction following dephospho-
rylation. Thus, the biosynthesis of laminitol and its
related epimer mytilitol in algae appears to proceed as
follows: D-mannoheptulose 7-P ? 1-deoxy-D-manno-
heptulose 7-P ?1-deoxy-D-gluco-heptulose 7-P ?
laminitol $ mytilitol (Woloszczuk and Hoffmann-
Ostenhof 1974). Condensation between C2 and C7 of L-
deoxy-D-gluco-heptulose, presumably as the 7-P
derivative, is believed to result in ring closure and
laminitol formation.
Compartmentalization of seven-carbon sugar
metabolism
Despite the prevalence and obvious importance of 7-C
sugars in many algae and vascular plant species, very
123

little is known about the compartmentalization and
metabolism of these monosaccharides. At least three
reaction sequences involving intermediates of both the
PPP and the PCR cycle have been proposed to
contribute to formation of the 7-C sugar precursor
Sed. Included are: (a) aldolase catalyzed conversion of
Ery 4-P ? dihydroxyacetone phosphate $ Sed 1,7-
bis-P; (b) TKT conversion of Xul 5-P ? Rib 5-P $
Sed 7-P ? Gly 3-P; and (c) TAL conversion of Fru
6-P ? Ery 4-P $ Sed 7-P ? Gly 3-P (Liu et al. 2002).
Fructose-bis-phosphate aldolase (FBA; Fru-bis-
phosphate aldolase, EC 4.1.2.13) is specific for its
substrate and is one of the non-regulated enzymes of
the PCR cycle (Uematsu et al. 2012). The enzyme has
the potential to control photosynthetic carbon flux
through the cycle, and overexpression in transgenic
plants enhances growth and increases biomass. In
contrast, TKT is involved in both the PPP and the PCR
cycle (Caillau and Quick 2005). While not all of the
enzymes of the PPP occur in the cytoplasm (Rocha
et al. 2014), TKT catalyzes two important reactions. In
the first reaction of the non-oxidative PPP, thiamine
diphosphate (TPP) accepts a 2-C fragment from D-Xul
5-P and transfers this fragment to D-Rib 5-P to form
Sed 7-P. The abstraction of two carbons from D-Xul 5-
P yields Gly 3-P. In the PCR cycle, TKT catalyzes the
reverse reaction, i.e. conversion of Sed 7-P and Gly 3-
P to D-Rib 5-P and D-Xul 5-P. The second reaction
catalyzed by TKT in the oxidative PPP also involves a
TPP-mediated transfer of a 2-C fragment from D-Xul
5-P to Ery 4-P, affording Fru 6-P and Gly 3-P. Again,
in the PCR cycle exactly the same reaction occurs but
in the opposite direction. Moreover, in photosynthesis
this is the first reaction catalyzed by TKT, rather than
the second. The final route to Sed is via TAL, which
transfers a 3-C fragment in the formation of Fru 6-P,
and is considered to be restricted to chloroplasts
(Schnarrenberger et al. 1995). Indeed, all putative
TKT and TAL genes in Arabidopsis contain a plastid-
targeting sequence (Eicks et al. 2002), indicating that
the gene products are confined to plastids in higher
plants. Metabolic flux quantification studies would
appear to support plastid localization of these enzymes
(Ganesh Sriram et al. 2004) and the emerging general
picture that TKT and TAL operate only in plastids
(Kruger and von Schaewen 2003). Even so, TAL
together with TKT provides a reversible link between
the PPP and glycolysis. Unlike TKT, which uses TPP
as a co-factor, TAL uses lysine to form Schiff base
123
Phytochem Rev
adducts with the ketose substrates (i.e. D-Fru 6-P and
D-Sed 7-P) and the acceptor aldoses (i.e. D-Ery4-P and
D-Gly 3-P) according to the following equilibrium:
Fru 6 - P þ Ery 4 - P $ Gly 3 - P þ Sed 7 - P
Structural characterization of the different TAL
reaction intermediates has been achieved using cry-
ocrystallography and has confirmed formation of the
Schiff base adducts between lysine and the donors Fru
6-P and Sed 7-P and the Michaelis complex with Ery
4-P (Lehwess-Litzmann et al. 2011).
Active photosynthesis stimulates formation of free
Sed in CAM plants, while elevated CO2 and exogenous
Suc enhance accumulation of this 7-C sugar (Ceusters
et al. 2013). Furthermore, studies using transgenic
antisense plants indicate that Sed 1,7-bis-phosphatase
(SBPase; EC 3.1.3.37), FBA, and TKT may increase
carbon flux through the PCR cycle to enhance produc-
tivity and yield (Raines 2011). Whether TAL influences
carbon flux similarly is currently unknown. Even so, the
SBPase loss-of-function mutant sbp of Arabidopsis, has
reduced growth due to inhibited cell division and
expansion (Liu et al. 2012), while Cucumis sativus
expressing an antisense TKT construct shows decreased
photosynthesis and yield (Bi et al. 2015). Accumulation
of Sed in source leaves of CAM plants might therefore
act to sequester excess carbon and mitigate Suc accu-
mulation and Suc-induced down regulation of photo-
synthesis (Ceusters et al. 2013). It was further argued that
this mechanism is flexible and also operates when sink
demand is saturated. So, partitioning of carbon from Suc
to Sed, presumably via Fru by TAL, occurs without
suppression of photosynthesis in source leaves. Like-
wise, in photosynthetically active sink tissues Suc
import, hydrolysis by INV and/or SuSy, and re-synthesis
via SPS might be expected to exert a similar effect to
delay or retard development. Since accumulation of Sed
in CAM species is considered an acclimation-avoiding
mechanism to prevent Suc accumulation, Pi depletion,
and suppression of photosynthetic carbon reduction,
formation and accumulation of 7-C heptuloses like
mannoheptulose, and coriose in other species, may fulfil
a similar purpose. Consequently, partitioning of reduced
carbon to a 7-C sugar(s) for storage by source and sink
tissue has the potential to sustain coordinated develop-
ment by providing flexibility and competitive advantage
to growing sinks.
Amounts of mannoheptulose in plants are reported
to be comparable to those of Sed (Okuda and Mori
Phytochem Rev
1974) and chemical dissection has revealed that
mannoheptulose is related to perseitol and volemitol
(Fig. 2) in the same manner as Sed is related to
volemitol and its epimer b-sedoheptitol (Richtmyer
1970a). Epimerization of laminitol to mytilitol is the
final step in the biosynthetic pathway of this 7-C
cyclitol in Chlorella fusca (Wöber and Hoffmann-
Ostenhof 1970a; Woloszczuk and Hoffmann-Osten-
hof 1974). Volemitol, its C3 epimer b-sedoheptitol,
and perseitol (the C4 epimer b-sedoheptitol) may
therefore be end products of 7-C sugar biosynthesis in
vascular plants. Both perseitol and volemitol occur
naturally (Richtmyer 1970b) and may be similarly
derived. Additionally, in species of Rosaceae, aldoses
(e.g. Glc) are metabolically related to alditols and
ketoses by NADPH?-reductase and NAD?-dehydro-
genase catalyzed interconversion (Aguayo et al.
2013). The presence in vascular plants of a putative
Sed-reductase responsible for volemitol formation
(Häfliger et al. 1999), and a putative aldoketose-
reductase for mannoheptulose-perseitol interconver-
sion (Tesfay et al. 2012), suggests that reaction
mechanisms involving isomerization and/or oxido-
reduction may be integral to 7-C sugar alcohol
biosynthesis. However, and as already discussed, the
origin of Sed 7-P and mannoheptulose 7-P as precur-
sors to heptoses and heptitols in plants is still unclear.
Mannoheptulose 7-P and mannoheptulose are read-
ily extracted from plant tissues (Datta and Racker
1961) and have been identified as products of exoge-
nously supplied CO2 (Nordal and Benson 1954; Liu
et al. 2002). Extensive studies in animal systems led to
the identification of D-Ara 5-P, D-mannoheptulose 7-P,
and D-Sed 1,7-bisP as products of [U-14C]-Rib 5-P
metabolism and demonstrated that the isomers, Sed
7-P and mannoheptulose 7-P, occur in all heptulose
7-P-containing fractions (Williams et al. 1978). Fur-
thermore, although D-Ara 5-P did not support CO2
fixation directly, in either leaves or chloroplasts from
Spinacia oleracea, it was shown to be derived from
D-glycero-D-ido-octulose 1,8-bisP and play an active
role in TAL and aldolase exchange reactions in the
reductive pentose pathway of carbon fixation (Wil-
liams and MacLeod 2006). The former represents the
first evidence of a D-Ara reaction in higher plant
chloroplasts. Since octulose phosphates show charac-
teristics of cyclic intermediates rather than synthe-
sized storage products (Williams and MacLeod 2006;
Flanigan et al. 2006), it is tempting to suggest that
under appropriate conditions and in competent
species, D-Ara acts as the aldopentose acceptor in
TPP-dependent TKT catalyzed formation of manno-
heptulose 7-P.
Interestingly, Ara 5-P is also a competitive inhibitor
of TAL reactions, with a Ki of 70 lM (Williams et al.
1978) while D-mannoheptulose 7-P, a product of TKT
action involving a ketose-P donor and Ara 5-P, is a
competitive inhibitor of TKT-catalyzed reactions
involving Ery 4-P (Williams and MacLeod 2006;
Flanigan et al. 2006).
Although the origin of D-Ara 5-P in plants has still to
be empirically ascertained, UDP-L-Ara is generated de
novo from UDP-D-Glc via UDP-D-glucuronate fol-
lowed by epimerization of UDP-D-Xul prior to incor-
poration into arabinosylated cell wall components
(Burget et al. 2003). Glycosyl hydrolases mediate cell
wall modification during expansion growth and catal-
yse the release of L-Ara from the polysaccharide-
containing cell wall matrix. As mentioned earlier, when
L-Ara was supplied to leaves of Medicago sativa it was
readily converted into L-gluco-heptulose, a precursor of
the algal 7-C cyclitol laminitol. Thus, heptitol forma-
tion may serve multiple functions in both source and
sink tissues and provide a mechanism to tolerate
fluctuations in the availability of resources for growth,
including sugar hydrolysis and reintegration during cell
wall assembly and disassembly.
Seven-carbon sugar transport
Among the benefits of synthesizing and accumulating
polyols including heptitols such as perseitol, volemi-
tol, and b-sedoheptitol are: provision of a highly
reduced carbon sink to aid in avoidance of sugar
mediated down regulation of photosynthesis; facili-
tating regeneration of NADP?/NAD? to allow for
continued PCR cycle activity thus protecting photo-
systems from oxidative damage; formation of stable,
chemically inert transportable forms of carbon that are
not readily metabolized and do not fluctuate signifi-
cantly in the short-term; and, as compatible solutes to
cope with low external osmotic potentials and promote
the hydration of membranes and proteins via prefer-
ential exclusion (Andersen et al. 2011; Merchant and
Richter 2011). Although the known heptitols are
currently without assigned function, characterization
of a perseitol-K? complex from Scurrula fusca leaves
123

(Ishizu et al. 2001) coupled with the ability of
perseitol, volemitol and other heptitols and heptoses
to form complexes with tungstate and molybdate
(Chapelle and Verchère 1991, 1995; Matulová et al.
1996; Chapelle et al. 1998) suggests a role for 7-C
sugars and sugar alcohols in metal ion chelation and
translocation.
Once produced, sugars are transported in the
phloem of vascular plants to sink organs (Lemoine
et al. 2013; Turgeon and Wolf 2009). Similarly, some
algae appear to translocate fixed carbon, primarily as
mannitol, and data from Macrocystis integrifolia has
confirmed the general picture for Laminariales that in
these organisms, translocation follows a source-sink
relationship similar to that described for higher plants
(Schmitz and Srivastava 1979). There is no published
information on transport of 7-C polyols in algae. For
higher plants, 7-C sugar transport has been most
studied in avocado trees. Sap collected from the
petioles of mature source leaves contains Suc, man-
noheptulose, and perseitol (Liu et al. 1999a, b, 2002).
Furthermore, sap collected from girdled branches
contained mannoheptulose that was unde-
tectable 7 days after girdling, whereas the perseitol
concentration remained unchanged (Tesfay et al.
2012). Xylem vessels are apparently unaffected by
girdling in avocado (Liu et al. 2002) which suggests
that mannoheptulose, in this species, is phloem
translocated. In another study it was shown that
phloem sap siphoned from the trunk (mobilized
carbohydrate reserves) and leaf petioles (photosyn-
thetically produced sugars) of avocado trees contained
in addition to mannoheptulose (27 %) and perseitol
(15 %), Suc, Glc and Fru in amounts 5 % or less of the
total soluble sugar component (Cowan 2004). Thus,
mannoheptulose and perseitol appear to be the major
mobile 7-C sugars in the avocado tree and presumably
both are available for growth and development. By
monitoring changes in fruit sugar content and compo-
sition over the course of development until har-
vestable maturity, the fate of tree-derived sugars could
be deduced (Cowan 2004). Fructose concentration in
young fruits declined from 22 % to about 4 % of the
total soluble sugars at harvestable maturity. Similarly,
Glc declined to less than 1 % of total soluble sugars.
The concentration of perseitol remained constant
throughout avocado fruit growth at about 25 %
whereas mannoheptulose increased from 15 % in
immature fruit to 53 % of the total soluble sugars in
123
Phytochem Rev
fruit near harvestable maturity. After harvest, and
during ripening, Suc, Glc, Fru, and mannoheptulose
declined, suggesting that these sugars are consumed as
part of the respiratory climacteric, which occurs
coincident with accumulation of triglycerides.
Although INV, SuSy and SPS activities have been
detected in avocado seed and fruit flesh (Richings et al.
2000), the contribution of these enzymes to 7-C sugar
homeostasis during fruit development is unknown.
In addition to imported sugar, growth and devel-
opment occurs coincident with acquisition of nutri-
ents. By way of example, in vascular plants,
cyanobacteria, algae, and fungi, boron (B) is essential
for cell wall formation, cell division and elongation,
membrane function, and carbohydrate metabolism and
transport (Bogiani et al. 2013; Goldbach and Wimmer
2007; Cong et al. 2015). While a major function of B is
the cross linking of cell wall pectin to facilitate normal
turgor-driven wall expansion (O’Neill et al. 2001) the
pectin glucuronyl-transferase 1 gene NpGUT1,
responsible for this cross-linking, is also required for
plant reproductive tissue development and fertiliza-
tion (Iwai et al. 2006). Boron deficiency in higher
plants, particularly during reproductive growth
directly affects productivity (Cong et al. 2015). The
reported decrease in carbohydrate production and
export from source tissues in response to B deficiency
leads to rapid inhibition of growth that presumably
arises due to decreased sink demand. For example,
Arabidopsis thaliana plants carrying the bor1 muta-
tion are acutely dwarfed (Noguchi et al. 1997). The
responsible gene was shown to encode a membrane-
localized protein that mediates export of B from
pericycle cells into the apoplast (Takano et al. 2002).
Thus, BOR1 appears to increase the B concentration of
xylem sap to supply and thus protect aerial plant parts
from B deficiency. Phloem translocation of B has also
been demonstrated for other species, typically in
association with sorbitol or mannitol (Cong et al.
2015; O’Neill et al. 2001), while the recently isolated
B-perseitol complex from avocado (Minchin et al.
2012) provides support for both phloem mobility of
this nutrient and a role for heptitols in B translocation.
In model systems like Arabidopsis thaliana, B
moves into the root via diffusion or active transport
mediated by NOD26-LIKE MAJOR INTRINSIC
PROTEIN5;1 (NIP5;1), a boric acid channel protein
(Takano et al. 2006). The efflux transporter BOR1,
modulated by B availability, then moves B into the
Phytochem Rev
xylem for transport to aerial plant parts. BOR1 is
involved not only in B transport into the xylem but also
B distribution to shoots and preferentially to young
leaves and presumably, other developing sinks such as
flowers and fruits. Boron remobilization occurs in
plants that form and export phloem-mobile borate-
polyol complexes, and the relative abundance of
polyols in the leaf phloem is a major factor determin-
ing the potential of B export and/or remobilisation
(Bellaloui et al. 2003). Partitioning of B between
xylem and phloem seems to be a function of NIP6;1, a
plasma membrane-localised channel protein that lacks
water transport activity (Miwa and Fujiwara 2010). In
Olea europaea (olive) the mannitol concentration of
leaf phloem sap has been suggested to promote B
remobilisation when external supply is inadequate
(Liakopoulos et al. 2009). A low or negligible external
supply of B to olive seedlings led to reduced amounts
of sap B, increased mannitol, and sustained high
remobilisation. These findings were interpreted as
indicative of a strategy by source leaves of olive to
remobilse B through depletion of the sap B pool.
Whether the B-perseitol complex of avocado reflects a
similar role in mediating export and remobilisation of
B from mature source leaves to sink tissues is
unknown. Even so, by monitoring xylem transport of
calcium and B to leaves in Arabidopsis, preferential
allocation and accumulation of B by young leaves was
demonstrated (O’Neill et al. 2001). Thus, redistribu-
tion of B from mature to young leaves in avocado
seems to mirror the pattern of carbohydrate export
from mature leaves and be a consequence of perseitol,
which complexes with B to facilitate phloem mobility
of this essential nutrient.
Another heptitol that is likely to play a role in metal
ion transport and redistribution is volemitol. As
already mentioned, volemitol, also present in avocado,
readily complexes both tungsten and molybdenum
(Mo) and is strongly labelled during photosynthetic
14
CO2 assimilation by mature source leaves of Prim-
ula (Kremer 1978). In addition, phloem exudate from
Primula was shown to contain both Suc and volemitol
(Häfliger et al. 1999). Although this heptitol does not
occur in all species of the genus Primula, sensitivity to
Mo deficiency by many species of Primula has long
been recognized and the requirement for Mo is greater
when prevailing conditions are cool (Reeker 1959). It
is thus not surprising perhaps that the volemitol
concentration of mature leaves from wild-grown
polyanthus plants was two-fold higher in March
(spring: flowering) than in June (summer: post flow-
ering) (Häfliger et al. 1999), when demand for Mo is
lower.
Studies on plant uptake and transport of Mo have
revealed the process to be rapid, controlled, acropetal
and basipetal, and sulfate-sensitive (Shinmachi et al.
2010; Bittner 2014). The discovery of Mo-specific
transporters such as MOT1 in algae (e.g. Chlamy-
domonas) and vascular plants (e.g. Arabidopsis
thaliana) coupled with the identification of MOT2, a
Mo transporter of the sulfate transporter family and its
localization to the vacuole, supports coordinated Mo
uptake and redistribution. Indeed, MOT2-deficient
plants show Mo accumulation in leaves and reduced
seed Mo concentration (Gasber et al. 2011). Whether
Mo redistribution mirrors the pattern of carbohydrate
movement and in particular volemitol in polyanthus is
currently unknown.
Integration of seven-carbon sugar metabolism
and transport
So far this review has focused attention on the
occurrence, metabolism, and transport of 7-C sugars
and sugar alcohols. Although the first 7-C sugar was
identified more than a century ago, it is evident from
the preceding discussion of published data that our
knowledge and understanding of 7-C sugar metabo-
lism and hence function is rudimentary. It seems that
production of 7-C sugars and heptitols is indeed
inextricably linked to primary metabolic processes
and that these carbohydrates contribute to the phys-
iology of the organisms in which they occur. Whereas
Sed and its related heptitol volemitol originate as
products of Sed 7-P, an intermediate of both the PCR
cycle and PPP, the origin of mannoheptulose and
perseitol has remained obscure. As already mentioned,
these two heptuloses differ in stereochemical config-
uration at C3-C4. In Sed, this configuration is threo
whereas in mannoheptulose it is erythro, which
strongly suggests that these monosaccharides are
formed independently. Interestingly, it has been
established that metal-heptitol (and heptose) com-
plexes in which the hydroxyl groups are in the erythro
(erythritol, galactitol) configuration (type E) such as
perseitol, are generally favoured (Chapelle and
Verchère 1991, 1995). Type E heptitol complexes
123

have also been characterized for ligands possessing an
arabino site of chelation e.g. volemitol. Thus, it is
proposed that a major physiological role of 7-C sugars
and heptitols, in addition to serving as carbon sinks,
probably involves metal ion chelation and transloca-
tion to fulfil nutrient demand.
Both Sed and mannoheptulose, like Suc, are
products of the reductive pentose phosphate pathway
in mature source leaves, and formation of 7-C sugars is
stimulated during active photosynthesis. Feeding of
Suc to isolated mature leaves of Kalanchoë pinnata
revealed modest accumulation of Suc and a 30-fold
increase in Sed after 3 days (Ceusters et al. 2013).
Similar results were reported for mature leaves of
Hylotelephium spectabile. Mature source leaves of
avocado incorporated CO2 into mannoheptulose 7-P,
mannoheptulose, Sed 7-P, Suc, and perseitol. Sed and
volemitol were also readily labelled following expo-
sure of mature polyanthus leaves to 14CO2 (Häfliger
et al. 1999).
There exists only one report on Suc application to a
7-C sugar-containing sink and in this example (Cowan
2004) radiolabel from Suc fed to developing avocado
fruit was readily incorporated into Glc, Fru and
mannoheptulose, but not perseitol. Furthermore, per-
seitol supplied via the transpiration stream to young
avocado fruit had little or no effect on endogenous
mannoheptulose content, whereas exogenous manno-
heptulose led to an increase in perseitol content
without significant impact on mannoheptulose con-
centration. This finding suggests that mannoheptulose
is a precursor of perseitol and that conversion to
perseitol occurs independent of photosynthesis.
Indeed, as long ago as 1962 it was reported by Bean
and co-workers (Bean et al. 1962) that Fru and
perseitol in avocado leaves were produced at the
expense of Suc and mannoheptulose respectively and,
by reactions separate from photosynthetic carbon
reduction. These authors proposed further that per-
seitol was formed from mannoheptulose, which in turn
was derived from Fru.
That leaf and fruit heptulose concentration
increases in response to exogenous Suc suggests that
Suc metabolism is linked to 7-C sugar and heptitol
biosynthesis. In plants, the metabolism of Suc involves
formation of free Glc and Fru, catalysed by INV and/or
formation of UDP-Glc and Fru catalysed by SuSy
(Koch 2004). Free hexose and hexulose are phospho-
rylated by different isoforms of HK and by
123
Phytochem Rev
fructokinase (FK; EC 2.7.1.4) to give the correspond-
ing hexose and hexulose phosphates. A substantial
proportion of hexose is partitioned into the Golgi for
cell wall glucan synthesis in actively growing organs.
Thus, Glc 6-P may enter either the glycolytic pathway
or be converted into UDP-Glc for synthesis of cell wall
glucan, depending on the energy demands of cells.
Fructose is also prevalent in plants following Suc
hydrolysis and has long been proposed as a potential
signalling molecule (Pego and Smeekens 2000). For
example, suppression of FK, responsible for the
formation of Fru 6-P, resulted in growth inhibition in
tomato (German et al. 2003) presumably due to Fru
accumulation. More recently, the function of Fru as a
regulatory metabolite was confirmed through its
interaction with abscisic acid and ethylene signalling,
and shown to induce developmental arrest in Ara-
bidopsis thaliana seedlings (Cho and Yoo 2011).
Thus, strategies to avoid the negative effects of Fru
accumulation likely exist in both source and sink
tissues. The carrier protein SWEET17 was identified
as controlling Fru content in Arabidopsis leaves
(Chardon et al. 2013). Accumulation of Fru following
reduced expression of SWEET17, coupled with local-
ization of this transporter protein to the tonoplast
provides ample support for vacuolar sequestration of
Fru. Operation of such strategies might also explain
the inability of early work that attempted to demon-
strate heptulose accumulation in leaves of Medicago
sativa fed D-Fru (Rendig and McComb 1964). Forma-
tion of heptuloses in competent species may be an
additional strategy to mitigate effects of Fru. Thus,
flux of carbon from Suc to Sed 7-P in CAM plants such
as Kalanchoë pinnata and Hylotelephium spectabile,
via Fru 6-P and the aldose acceptor D-Ery 4-P,
catalyzed by a TAL, mitigates Suc-induced suppres-
sion of photosynthesis to sustain growth and develop-
ment. In avocado, formation of mannoheptulose from
Suc presumably involves the conversion of Fru 6-P by
TPP-dependent TKT action involving the aldopentose
acceptor D-Ara, to fulfil a similar purpose. It is thus
distinctly possible that TAL and TKT convert Fru 6-P
to Sed 7-P and mannoheptulose 7-P simultaneously in
competent tissues of 7-C sugar containing plants.
Since decreased TKT activity inhibits the regeneration
of ribulose-1,5-bisP and reduces photosynthesis
(Henkes et al. 2001; Satoh et al. 2004), conversion
of heptuloses to the corresponding heptitols may in
deed represent part of an integrated mechanism to
Phytochem Rev
sustain carbon assimilation. This is achieved by
enhanced carbon and P homeostasis, and providing a
pool of polyols as a carbon sink and for nutrient
acquisition.
Heptuloses and heptitols are phloem mobile, how-
ever, unlike the situation in avocado, Suc rather than
the heptitol dominates in phloem exudate of polyan-
thus. Also, the carbohydrate composition of polyan-
thus phloem sap is distinct from that of leaf tissue, in
which volemitol dominates at 71 % of the total soluble
leaf carbohydrate. One possible explanation for this
difference is that source leaves of polyanthus also
function as volemitol sinks and that a strategy similar
to that for B-mannitol complex remobilization in Olea
europaea operates in this species. Thus, volemitol
accumulation in source leaves of polyanthus could be
a consequence of remobilization after formation of a
metal-heptitol chelate(s) to fulfil nutrient demands.
Other important minerals that may be transported as
metal-heptitol complexes include potassium, calcium,
magnesium, iron, and B. Indeed, Primulas are
extremely sensitive to iron (Fe) availability and are
considered an indicator crop for insufficient soil Fe
(Karlsson 2001). In this regard it is noteworthy that Fe
availability appears intimately related to plant Mo
metabolism, and that crosstalk between Fe and Mo
exists for Mo cofactor (Moco) biosynthesis and
assembly (Bittner 2014). Moco-requiring enzymes
are involved in nitrogen assimilation, purine degrada-
tion, phytohormone synthesis, and sulfite detoxifica-
tion. In plants some of these enzymes are regulated by
sugar via the protein kinase, sucrose non-fermenting-
1-related protein kinase 1 (Li et al. 2009).
Proposed pathway of seven-carbon sugar
biosynthesis
The accumulated information presented here indicates
that heptoses, heptuloses, and heptitols occur in many
photosynthetic organisms (i.e. plants and algae), fungi,
and bacteria. Studies on the biosynthesis of C-7 sugars
support the notion that these carbohydrates arise from
CO2 in photosynthetic organisms. However, the
precise pathway of formation remains to be elucidated.
Evidence suggests that Sed 7-P and mannoheptulose
7-P are precursors to Sed and mannoheptulose
respectively, but the origin of the phosphorylated
precursors is unknown. Potential routes include TAL
Fig. 4 Proposed pathway for the biosynthesis of 7-C sugars
from D-Fru 6-P. Suc-derived Fru 6-P activates a TKT to give Ery
4-P and the corresponding a-hydroxyacetyl (a-OHAc) moiety.
Erythrose 4-P accepts the dihydroxyacetone (DHAc) group
from Fru 6-P, catalyzed by TAL with the release of Gly 3-P, to
yield Sed 7-P. Both Sed 7-P and mannoheptulose 7-P are
substrates for TKT and yield the a-OHAc acceptors, Rib 5-P and
Ara 5-P
catalyzed condensation of a dihydroxyacetone moi-
ety from D-Fru 6-P with the aldose D-Ery 4-P to form
Sed 7-P, and TKT-catalyzed transfer an a-hydroxy-
acetyl moiety from D-Fru 6-P to the aldopentose
acceptor D-Ara 5-P for mannoheptulose 7-P formation.
Although confirmation of these reaction sequences is
still required, as is the contribution of both the non-
oxidative PPP and PCR cycle in the supply of the
ketose donors and aldose acceptors, the scheme illus-
trated in Fig. 4 is proposed for 7-C sugar biosynthesis
from D-Fru 6-P.
It is hypothesized that a TKT-dependent non-
oxidative pentose phosphate shunt-type mechanism
contributes to 7-C sugar biosynthesis. In summary,
accumulation of Fru 6-P in either source or sink tissues
of competent species is the donor ketose to support
heptulose-synthesizing TKT activity. This is achieved
by activation of a TKT-dependent shunt to give Ery
4-P and the corresponding a-hydroxyacetyl moiety.
Erythrose 4-P is the acceptor of a 3-C dihydroxyace-
tone group from Fru 6-P, catalyzed by TAL with the
release of Gly 3-P, to yield Sed 7-P. Both Sed 7-P and
mannoheptulose 7-P are substrates for TKT and yield
the a-hydroxyacetyl acceptors, Rib 5-P and Ara 5-P. In
algae, Ara is also the 2-C acceptor for biosynthesis of
D-mannoheptulose 7-P, 1-deoxy-D-gluco-heptulose
7-P and its corresponding polyol laminitol (Woloszc-
zuk and Hoffmann-Ostenhof 1974). Thus, continued
flux from Fru 6-P via the TKT-dependent heptulose
123

shunt results in accumulation of Sed 7-P and/or
mannoheptulose 7-P which are the respective precur-
sors to free Sed and mannoheptulose, and the corre-
sponding polyols, volemitol and perseitol. Support for
the operation of a TKT-dependent shunt has been
forthcoming from studies in which, Panc-1 pancreatic
cancer cells supplied D-Fru, revealed a 200 % increase
in TKT protein expression and a 280 % increase in
enzyme activity (Liu et al. 2010). Additionally, it was
demonstrated by metabolomic study that D-Fru was
preferentially metabolized in cancer cells to Rib by
TKT-regulated, non-oxidative PPP and that this
increased flux into nucleic acids occurred coincident
with elevated uric acid. Interestingly, the latter is
derived from hypoxanthine and xanthine in a reaction
sequence catalyzed by xanthine dehydrogenase (EC
1.17.1.4), a well-known Mo-requiring enzyme (Men-
del and Kruse 2012).
Operation of a plant TKT-dependent heptulose
shunt might also suggest that Sed 7-P and mannohep-
tulose 7-P arise simultaneously and that these sugar
phosphates should be produced in comparable
amounts. However, Ara 5-P a product of TKT action
on mannoheptulose 7-P is an inhibitor of TAL activity
(Caillau and Quick 2005; Williams et al. 1978).
Indeed, Ara 5-P like Fru 6-P forms a covalent Schiff
base at the TAL active site preventing displacement of
a water molecule, which causes conformational
change and altered enzyme reactivity (Light and
Anderson 2014). Inhibition of TAL by Ara 5-P,
coupled with mannoheptulose 7-P inhibition of TKT
action on Ery 4-P (Williams et al. 1978; Flanigan et al.
2006), would therefore be expected to reduce flux of
Fru 6-P to Sed 7-P in favour of mannoheptulose 7-P in
mannoheptulose-accumulating species such as avo-
cado. In leaves of the Sed-accumulating CAM plant,
Kalanchoë pinnata, Sed arises from Sed 7-P by action
of a Sed 7-P phosphatase (Ceusters et al. 2013).
Moreover, activity of Sed 7-P phosphatase was
20-fold higher in leaves of Suc-induced stems. Thus,
accumulation of Sed in this species appears to be
regulated at the level of substrate supply, a primary
role of the proposed TKT-dependent heptulose shunt.
Generation of phosphorylated Sed and mannohep-
tulose provides a pool of heptulose for in situ synthesis
of the respective heptitols. Both Sed and mannohep-
tulose reductase activities have been detected in
heptulose accumulating species (Häfliger et al. 1999;
Tesfay et al. 2012) and, these heptuloses and the
123
Phytochem Rev
corresponding heptitols are phloem mobile. Non-
photosynthetic sink tissues such as roots and seeds
also contain heptoses and these are likely formed by
isomerization following import of the corresponding
phloem translocated heptulose. Thus, D-glycero-D-
gluco-Hep is formed by isomerization of Sed in roots
of primula whereas mannoheptulose is isomerized to
D-glycero-D-galacto-Hep in seed of avocado.
Conclusion
Since the discovery of 7-C sugars more than 100 years
ago significant progress has been made in carbohy-
drate research. It is now accepted that sugar signals
play a central regulatory function in plant growth and
development. This overview represents the first com-
prehensive synthesis of knowledge about the occur-
rence, metabolism, transport and function of 7-C
sugars, a relatively underexplored area of carbohy-
drate biology that needs to be integrated into main-
stream sugar research.
It is apparent from the literature surveyed that 7-C
sugars and sugar alcohols occur in many plants, algae,
fungi, and bacteria and that formation of a heptulose
monophosphate, in most cases Sed 7-P, represents the
committed step in the biosynthesis of these monosac-
charides. Until now, Sed 7-P was considered to arise
either from the PPP or the PCR cycle or both.
However, a re-evaluation of published data strongly
suggests that 7-C sugar biosynthesis originates from
Fru 6-P generated following hydrolysis of Suc via a
TKT-dependent heptulose shunt. A recently published
transcriptome analysis of mesocarp (fruit flesh) from
developing fruit of avocado, a species with fruits and
seeds well known to accumulate large quantities of
mannoheptulose and perseitol, has revealed complete
glycolytic pathways both in plastids and cytoplasm of
this sink tissue; that transcripts of SuSy and vacuolar
INV are highly expressed throughout fruit develop-
ment; and, that a high level of expression of TKT and
TAL orthologs occurs in fruit mesocarp plastids
(Kilaru et al. 2015). Although a major objective of
this transcriptome study was to determine which genes
associated with lipid biosynthesis are predominantly
expressed, and how expression patterns vary with fruit
developmental stage, the results also point to involve-
ment of both SuSy and INV in the supply of Fru for
7-C sugar biosynthesis. Thus, conversion of Suc-
Phytochem Rev
derived Fru 6-P to Sed 7-P and mannoheptulose 7-P in
competent tissues and species of 7-C sugar containing
organisms by TAL and TKT appears to be the major
pathway of 7-C sugar biosynthesis.
It is proposed that the physiological function of 7-C
sugars and heptitols, in addition to serving as carbon
sinks, involves metal ion chelation, translocation and
remobilization to fulfil nutrient demand. Indeed, the
apparent role of heptuloses and heptitols in transloca-
tion of B and Mo, and the suggestion that crosstalk
between Fe and Mo prevails during Moco biosynthesis
and assembly, would seem to support an inextricable
role for 7-C sugars in growth and development of
many plants.
Acknowledgments Financial support from the National
Research Foundation, Pretoria, South Africa (IFR
1202220169) is gratefully acknowledged. Professor Emeritus
Nigel Wolstenholme is thanked for critical reading and valuable
comments.
Compliance with ethical standards
Conflicts of interest The author states no conflict of interest
and has received no payment for the preparation of this
manuscript.
References
Aguayo MF, Ampuero D, Mandujano P, Parada R, Muñoz R,
Gallart M, Altabella T, Cabrera R, Stange C, Handford M
(2013) Sorbitol dehydrogenase is a cytosolic protein
required for sorbitol metabolism in Arabidopsis thaliana.
Plant Sci 205–206:63–75
Andersen HD, Wang CH, Arleth L, Peters GH, Westh P (2011)
Reconciliation of opposing views on membrane-sugar
interactions. Proc Natl Acad Sci USA 108:1874–1878
Angyal SJ, Le Fur R (1984) The 13C-N.M.R. spectra and the
conformation of heptitols in solution. Carbohydr Res
126:15–26
Bean RC, Barr BK, Welch HV, Porter GG (1962) Carbohydrate
metabolism in avocado. 1. Relations between sugars in
leaves during photosynthesis and subsequent dark periods.
Arch Biochem Biophys 96:524–529
Begbie R, Richtmyer NK (1966) The isolation of some heptoses,
heptuloses, octuloses, and nonuloses from Primula offici-
nalis Jacq. Carbohydr Res 2:272–288
Bellaloui N, Yadavc RC, Chern M-S, Hu H, Gillen AM, Greve
C, Dandekar AM, Ronald PC, Brown PH (2003) Trans-
genically enhanced sorbitol synthesis facilitates phloem-
boron mobility in rice. Physiol Plant 117:79–84
Bi H, Dong X, Wu G, Wang M, Ai X (2015) Decreased TK
activity alters growth, yield and tolerance to low temper-
ature and low light intensity in transgenic cucumber plants.
Plant Cell Rep 34:345–354
Bianchi G, Gamba A, Murelli C, Salamini F, Bartels D (1991)
Novel carbohydrate metabolism in the resurrection plant
Craterostigma plantagineum. Plant J 1:355–359
Bihmidine S, Hunter CT III, Johns CE, Koch KE, Braun DM
(2013) Regulation of assimilate import into sink organs:
update on molecular drivers of sink strength. Front Plant
Sci 4:177. doi:10.3389/fpls.2013.00177
Bittner F (2014) Molybdenum metabolism in plants and cross
talk to iron. Front Plant Sci 28:5. doi:10.3389/fpls.2014.
00028
Board M, Colquhoun A, Newsholme EA (1995) High Km glu-
cose-phosphorylating (glucokinase) activities in a range of
tumor cell lines and inhibition of rates of tumor growth by
the specific enzyme inhibitor mannoheptulose. Cancer Res
55:3278–3285
Bogiani JC, Amaro ACE, Rosolem CA (2013) Carbohydrate
production and transport in cotton cultivars grown under
boron deficiency. Sci Agric 70:442–448
Burget EG, Verma R, Mølhøj M, Reiter W-D (2003) The
biosynthesis of L-arabinose in plants: molecular cloning
and characterization of a Golgi-localized UDP D-xylose
4-epimerase encoded by the MUR4 gene of Arabidopsis.
Plant Cell 15:523–531
Caillau M, Quick WP (2005) New insights into plant transal-
dolase. Plant J 43:1–16
Ceusters J, Godts C, Peshev D, Vergauwen R, Dyubankova N,
Lescrinier E, De Proft MP, Van den Ende W (2013)
Sedoheptulose accumulation under CO2 enrichment in
leaves of Kalanchoë pinnata: a novel mechanism to
enhance C and P homeostasis? J Exp Bot 64:1497–1507
Chapelle S, Verchère J-F (1991) A 13C-n.m.r. study of the
tungstate and molybdate complexes of perseitol, galactitol,
and D-mannitol. Carbohydr Res 211:279–286
Chapelle S, Verchère J-F (1995) A 13C and 183W NMR study of
the structures of tungstate and molybdate complexes of
volemitol. Carbohydr Res 266:161–170
Chapelle S, K} oll P, Verchère J-F (1998) A multinuclear NMR
spectroscopy characterization of dinuclear tungsten(VI)
complexes of tridentate and pentadentate meso-D-glycero-
D-gulo-heptitol and D-glycero-L-gulo-heptitol. Carbohydr
Res 306:27–34
Chardon F, Bedu M, Calenge F, Klemens PAW, Spinner L,
Clement G, Chietera G, Léran S, Ferrand M, Lacombe B,
Loudet O, Dinant S, Bellini C, Neuhaus HE, Daniel-Vedele
F, Krapp A (2013) Leaf fructose content is controlled by
the vacuolar transporter SWEET17 in Arabidopsis. Curr
Biol 23:697–702
Charlson AJ, Richtmyer NK (1960) The isolation of an octulose
and octitol from natural sources: D-glycero-D-manno-oc-
tulose and D-erythro-D-galacto-octitol from the avocado
and D-glycero-D-manno-octulose from Sedum species.
J Am Chem Soc 82:3428–3434
Cho Y-H, Yoo S-D (2011) Signaling role of fructose mediated
by FINS1/FBP in Arabidopsis thaliana. PLoS Genet
7(1):e1001263. doi:10.1371/journal.pgen.1001263
Cong X, Jing H, Lin N, Xia N, Huang M, Jiang X (2015) Boron
deficiency affects cell morphology and structure of young
leaves of radish. Acta Physiol Plant 37:247. doi:10.1007/
s11738-015-2004-7
Courtois P, Sener A, Malaisse WJ (2000) Effects of D-manno-
heptose and D-glycero-D-gluco-heptose upon D-glucose
123

metabolism and insulinotropic action in rat pancreatic
islets and D-glucose phosphorylation by hexokinase
isoenzymes: comparison with D-mannoheptulose. Intl J
Mol Med 6:171–175
Cowan AK (2004) Metabolic control of avocado fruit growth:
3-hydroxy-3-methylglutaryl coenzyme A reductase, active
oxygen species and the role of C7 sugars. S Afr J Bot
70:75–82
da Silva MdeL, Gorin PA, Iacomini M (1993) Unusual carbo-
hydrates from the lichen, Parmotrema cetratum. Phyto-
chemistry 34:715–717
Dai N, Schaffer A, Petreikov M, Shahak Y, Giller Y, Ratner K,
Levine A, Granot D (1999) Overexpression of Arabidopsis
hexokinase in tomato plants inhibits growth, reduces
photosynthesis, and induces rapid senescence. Plant Cell
11:1253–1266
Datta AG, Racker E (1961) Mechanism of action of transketo-
lase. 1. Properties of the crystalline yeast enzyme. J Biol
Chem 236:617–623
Doidy J, Grace E, Kühn C, Simon-Plas F, Casieri L, Wipf D
(2012) Sugar transporters in plants and in their interactions
with fungi. Trends Plant Sci 17:413–422
Dvash L, Afik O, Shafir S, Schaffer A, Yeselson Y, Dag A,
Landau S (2002) Determination by near-infrared spec-
troscopy of perseitol used as a marker for the botanical
origin of avocado (Persea americana Mill.) honey. J Agric
Food Chem 50:5283–5287
Eicks M, Maurino V, Knappe S, Flügge UI, Fischer K (2002)
The plastidic pentose phosphate translocator represents a
link between the cytosolic and the plastidic pentose phos-
phate pathways in plants. Plant Physiol 128:512–522
Fiki AE, Metabteb GE, Bellebria C, Wartmann T, Bode R,
Gellissen G, Kunze G (2009) The Aruxula adeninivorans
ATAL gene encoding transaldolase—gene characteriza-
tion and biotechnological exploitation. Appl Micribiol
Biotechnol 74:1291–1299
Flanigan IL, MacLeod JK, Williams JF (2006) A re-investiga-
tion of the path of carbon in photosynthesis utilizing GC/
MS methodology. Unequivocal verification of the partici-
pation of octulose phosphates in the pathway. Photosynth
Res 90:149–159
Fukumoto T, Kano A, Ohtani K, Yamasaki-Kokudo Y, Kim
B-G, Hosotani K, Saito M, Shirakawa C, Tajima S, Izumori
K, Ohara T, Shigematsu Y, Tanaka K, Ishida Y, Nishizawa
Y, Tada Y, Ichimura K, Gomi K, Akimitsu K (2011) Rare
sugar D-allose suppresses gibberellin signaling through
hexokinase-dependent pathway in Oryza sativa L. Planta
234:1083–1095
Fukumoto T, Kano A, Ohtani K, Inoue M, Yoshihara A, Izumori
K, Tajima S, Shigematsu Y, Tanaka K, Ohkouchi T, Ishida
Y, Nishizawa Y, Tada Y, Ichimura K, Gomi K, Yoo S-D,
Sheen J, Akimitsu K (2013) Phosphorylation of D-allose by
hexokinase involved in regulation of OsABF1 expression for
growth inhibition in Oryza sativa L. Planta 237:1379–1391
Ganesh Sriram D, Fulton B, Iyer VV, Peterson JM, Zhou R,
Westgate ME, Spalding MH, Shanks JV (2004) Quantifi-
cation of compartmented metabolic fluxes in developing
soybean embryos by employing biosynthetically directed
fractional 13C labeling, two-dimensional [13C,1H] nuclear
magnetic resonance, and comprehensive isotopomer bal-
ancing. Plant Physiol 136:3034–3057
123
Phytochem Rev
Gasber A, Klaumann S, Trentmann O, Trampczynska A, Cle-
mens S, Schneider S, Sauer N, Feifer I, Bittner F, Mendel
RR, Neuhaus HE (2011) Identification of an Arabidopsis
solute carrier critical for intracellular transport and inter-
organ allocation of molybdate. Plant Biol (Stuttg)
13:710–718
German MA, Dai N, Matsevitz T, Hanael R, Petreikov M,
Bernstein N, Ioffe M, Shahak Y, Schaffer AA, Garnot D
(2003) Suppression of fructokinase encoded by LeFRK2 in
tomato stem inhibits growth and causes wilting of young
leaves. Plant J 34:837–846
Goldbach HE, Wimmer MA (2007) Boron in plants and ani-
mals: Is there a role beyond cell-wall structure? J Plant Nutr
Soil Sci 170:39–48
Häfliger B, Kindhauser E, Keller F (1999) Metabolism of D-
glycero-D-manno-heptitol, volemitol, in Polyanthus. Dis-
covery of a novel ketose reductase. Plant Physiol
119:191–197
Henkes S, Sonnewald U, Badur R, Flachmann R, Stitt M (2001)
A small decrease of plastid transketolase activity in anti-
sense tobacco transformants has dramatic effects on pho-
tosynthesis and phenylpropanoid metabolism. Plant Cell
13:535–551
Huang H, Rong H, Li X, Tong S, Zhu Z, Niu L, Teng M (2008)
The crystal structure and identification of NQM1/
YGRO43C, and transaldolase from Saccharomyces cere-
visiae. Proteins Struct Funct Genet 73:1076–1081
Ish Am G, Eisikowitch D (1998) Low attractiveness of avocado
(Persea americana Mill.) flowers to honey bees (Apis
mellifera L.) limits fruit set in Israel. J Hortic Sci
Biotechnol 73:195–204
Ishizu T, Tsujino E, Winarno H, Ohashi K, Shibuya H (2001) A
complex of perseitol and K? ion from Scurrula fusca
(Loranthaceae). Tetrahedron Lett 42:6887–6889
Iwai H, Hokura A, Oishi M, Ishii T, Sakai S, Satoh S (2006) The
gene responsible for borate cross-linking of pectin
rhamnogalacturonan-II is required for plant reproductive
tissue development and fertilization. Proc Natl Acad Sci
USA 103:16592–16597
Jeong J, Huber DJ (2004) Suppression of avocado (Persea
americana Mill.) fruit softening and changes in cell wall
matrix polysaccharides and enzyme activities: differential
responses to 1-MCP and delayed ethylene application.
J Am Soc Hort Sci 129:752–759
Karlsson MG (2001) Primula culture and production.
HortTechnol 11:627–635
Kilaru A, Cao X, Dabbs PB, Sung H-J, Rahman MdM, Thrower
N, Zynda G, Podicheti R, Ibarra-Laclette E, Herrera-
Estrella L, Mockaitis K, Ohlrogge JB (2015) Oil biosyn-
thesis in a basal angiosperm: transcriptome analysis of
Persea americana mesocarp. BMC Plant Biol 15:203.
doi:10.1186/s12870-015-0586-2
Koch K (2004) Sucrose metabolism: regulatory mechanisms
and pivotal roles in sugar sensing and plant development.
Curr Opin Plant Biol 7:235–246
Köll P, Komander H, Angyal SJ (1991) Crystal and molecular
structures of D-glycero-D-manno-heptitol (a-sedoheptitol,
volemitol) and D-glycero-D-gluco-heptitol (b-sedohepti-
tol). Carbohydr Res 218:55–62
Kourtoglou E, Mamma D, Topakas E, Christakopoulos P (2008)
Purification, characterization and mass spectrometric
Phytochem Rev
sequencing of transaldolase from Fusarium oxysporum.
Process Biochem 43:1094–1101
Kremer BP (1977) Biosynthesis of polyols in Pelvetia
canaliculata. Z Pflanzenphysiol 81:68–73
Kremer BP (1978) Volemitol in the genus Primula: distribution
and significance. Z Pflanzenphysiol 86:453–461
Kruger NJ, von Schaewen A (2003) The oxidative pentose
phosphate pathway: structure and organization. Curr Opin
Plant Biol 6:236–246
LaForge FB (1916) D-Mannoheptulose, a new sugar from the
avocado. J Biol Chem 28:511–522
LaForge FB, Hudson CS (1917) Sedoheptulose, a new sugar
from Sedum spectabile. J Biol Chem 30:61–77
Lehwess-Litzmann A, Neumann P, Parthier C, Lüdtke S, Golbik
R, Ficner R, Tittmann K (2011) Twisted Schiff base
intermediates and substrate locale revise transaldolase
mechanism. Nature Chem Biol 7:678–684
Lemoine R, La Camera S, Atanassova R, Dédaldéchamp F,
Allario T, Pourtau N, Bonnemain J-L, Laloi M, Coutos-
Thévenot P, Maurousset L, Faucher M, Girousse C,
Lemonnier P, Parrilla J, Durand M (2013) Source-to-sink
transport of sugar and regulation by environmental factors.
Front Plant Sci 4:272. doi:10.3389/fpls.2013.00272
Lewis DH, Smith DC (1967) Sugar alcohols (polyols) in fungi
and green plants. I. Distribution, physiology and metabo-
lism. New Phytol 66:143–184
Li X-F, Li Y-J, An Y-H, Xiong L-J, Shao X-H, Wang Y, Sun Y
(2009) AKINb1 is involved in the regulation of nitrogen
metabolism and sugar signaling in Arabidopsis. J Integr
Plant Biol 51:513–520
Liakopoulos G, Stavrianakou S, Nikolopoulos D, Karvonis E,
Vekkos K-A, Psaroudi V, Karabourniotis G (2009) Quan-
titative relationships between boron and mannitol con-
centrations in phloem exudates of Olea europaea leaves
under contrasting boron supply conditions. Plant Soil
323:177–186
Light SH, Anderson WF (2014) Arabinose 5-phosphate cova-
lently inhibits transaldolase. J Struct Funct Genomics
15:41–44
Liu X, Sherman G, Robinson PW, Witney GW, Arpaia ML
(1995) Nectar sugar composition of selected avocado cul-
tivars and related species. Subtrop Fruit News 3:8–9
Liu X, Robinson PW, Madore MA, Witney GW, Arpaia ML
(1999a) ‘Hass’ avocado carbohydrate fluctuations.
I. Growth and phenology. J Am Soc Hortic Sci
124:671–675
Liu X, Robinson PW, Madore MA, Witney GW, Arpaia ML
(1999b) ‘Hass’ avocado carbohydrate fluctuations. II. Fruit
growth and ripening. J Am Soc Hortic Sci 124:676–681
Liu X, Sievert J, Arpaia ML, Madore MA (2002) Postulated
physiological roles of the seven-carbon sugars, manno-
heptulose, and perseitol in avocado. J Am Soc Hortic Sci
127:108–114
Liu H, Huang D, McArthur DL, Boros LG, Nissen N, Heaney
AP (2010) Fructose induces transketolase flux to promote
pancreatic cancer growth. Cancer Res 70:6368–6376
Liu X-L, Yu H-D, Guan Y, Li J-K, Guo F-Q (2012) Carbony-
lation and loss-of-function analyses of SBPase reveal its
metabolic interface role in oxidative stress, carbon assim-
ilation, and multiple aspects of growth and development in
Arabidopsis. Mol Plant 5:1082–1099
Matulová M, Verchère J-F, Chapelle S (1996) Furanose vs.
acyclic forms of carbohydrate ligands. A multinuclear
NMR spectroscopy study of the molybdate and tungstate
complexes of D-glycero-L-manno-heptose. Carbohydr Res
287:37–48
McArthur F, Andersson CE, Loutet S, Mowbray SL, Valvano
MA (2005) Functional analysis of the glycero-manno-
heptose 7-phosphate kinase domain from the bifunctional
HldE protein, which is involved in ADP-L-glycero-D-
manno-heptose biosynthesis. J Bacteriol 187:5292–5300
McComb EA, Rendig VV (1961) Isolation and identification of
L-galacto-heptulose from plants fed L-sorbose. Arch Bio-
chem Biophys 95:316–319
Mendel RR, Kruse T (2012) Cell biology of molybdenum in
plants and humans. Biochim Biophys Acta
1823:1568–1579
Merchant A, Richter AA (2011) Polyols as biomarkers and
bioindicators for 21st century plant breeding. Funct Plant
Biol 38:934–940
Minchin PEH, Thorp TG, Boldingh HL, Gould N, Cooney JM,
Negm JB, Focht E, Arpaia ML, Hu H, Brown P (2012) A
possible mechanism for phloem transport of boron in
‘Hass’ avocado (Persea americana Mill.) trees. J Hortic
Sci Biotechnol 87:23–28
Miwa K, Fujiwara T (2010) Boron transport in plants: co-ordi-
nated regulation of transporters. Annal Bot 105:1103–1108
Morrison JP, Read JA, Coleman WG Jr, Tann ME (2005) Dis-
mutase activity of ADP-L-glycero-D-manno-heptose
6-epimerase: evidence for a direct oxidation/reduction
mechanism. Biochemistry 44:5907–5915
Noguchi K, Yasumori M, Imai T, Naito S, Matsunaga T, Oda H,
Hayashi H, Chino M, Fujiwara T (1997) bor1-1, an Ara-
bidopsis thaliana mutant that requires a high level of
boron. Plant Physiol 115:901–906
Noiraud N, Maurousset L, Lemoine RM (2001) Transport of
polyols in higher plants. Plant Physiol Biochem
39:717–728
Nordal A, Benson AA (1954) Isolation of mannoheptulose and
identification of its phosphate in avocado leaves. J Am
Chem Soc 76:5054–5055
O’Hara LE, Paul MJ, Wingler A (2013) How do sugars regulate
plant growth and development? New insight into the role of
trehalose-6-phosphate. Mol Plant 6:261–274
O’Neill MA, Eberhard S, Albersheim P, Darvill AG (2001)
Requirement of borate cross-linking of cell wall rhamno-
galacturonan II for Arabidopsis growth. Science
294:846–849
Ogata JN, Kawano Y, Bevenue A, Casarett LJ (1972) Keto-
heptose content of some tropical fruits. J Agric Food Chem
20:113–115
Okuda T, Mori K (1974) Coriose and related compounds. 5.
Distribution of manno-heptulose and sedoheptulose in
plants. Phytochemistry 13:961–964
Onishi H, Perry MB (1965) The production of meso-glycero-
ido-heptitol and D-glycero-D-ido-heptitol by Pichia meso.
Can J Microbiol 11:929–934
Onishi H, Perry MB (1972) The production of D-glycero-D-
manno-heptitol by Torulopsis versatilis. Can J Microbiol
18:925–927
Pego JV, Smeekens S (2000) Plant fructokinases: a sweet family
get together. Trends Plant Sci 5:531–536
123

Peshev D, Van den Ende W (2013) Sugars as antioxidants in
plants. In: Tuteja N (ed) Crop improvement under adverse
conditions. Springer, New York, pp 285–308
Pfetzing J, Stengel DB, Cuffe MM, Savage AV, Guiry MD
(2000) Effects of temperature and prolonged emersion on
photosynthesis, carbohydrate content and growth of the
brown intertidal alga Pelvetia canaliculata. Bot Mar
43:399–407
Porchia AC, Salerno GL (1996) Sucrose biosynthesis in a
prokaryotic organism: presence of two sucrose-phosphate
synthases in Anabaena with remarkable differences com-
pared with the plant enzymes. Proc Natl Acad Sci USA
93:13600–13604
Quintanilla-Licea R, Morado-Castillo R, Gomez-Flores R,
Laatsch H, Verde-Star MJ, Hernández-Martı́nez H,
Tamez-Guerra P, Tamez-Guerra R, Rodrı́guez-Padil C
(2012) Bioassay-guided isolation and identification of
cytotoxic compounds from Gymnosperma glutinosum
leaves. Molecules 17:11229–11241
Radakovits R, Jinkerson RE, Darzins A, Posewitz MC (2010)
Genetic engineering of algae for enhanced biofuel pro-
duction. Eukaryot Cell 9:486–501
Raines CA (2011) Increasing photosynthetic carbon assimila-
tion in C3 plants to improve crop yield: current and future
strategies. Plant Physiol 155:36–42
Reeker R (1959) Die Verhiitung des Molybdanmangels bei
Verwendung von Torf als Substrat. Gartenbauwissenschaft
24:528–545; Torfnachrichten (1960) Sonderdruck zu 11:5/6
Rendig VV, McComb EA (1962a) Synthesis of L-gluco-heptu-
lose in plants fed L-arabinose. Arch Biochem Biophys
97:562–564
Rendig VV, McComb EA (1962b) Pentoses as precursors of
heptuloses in plants. Arch Biochem Biophys 99:409–413
Rendig VV, McComb EA (1964) Steric specificity in synthesis
of heptuloses by plants. Plant Physiol 39:793–798
Rennie EA, Turgeon R (2009) A comprehensive picture of
phloem loading strategies. Proc Natl Acad Sci USA
106:14162–14167
Richings EW, Cripps RF, Cowan AK (2000) Factors affecting
‘Hass’ avocado fruit size: carbohydrate, abscisic acid and
isoprenoid metabolism in normal and phenotypically small
fruit. Physiol Plant 109:81–89
Richtmyer NK (1970a) The isolation of volemitol and other
polyhydric alcohols from avocado seed. Carbohydr Res
12:135–138
Richtmyer NK (1970b) The isolation of perseitol and volemitol
from Sedum, and some other observations on Sedum con-
stituents. Carbohydr Res 12:139–142
Rocha AG, Mehlmer N, Stael S, Mair A, Parvin N, Chigri F,
Teige M, Vothknecht UC (2014) Phosphorylation of Ara-
bidopsis transketolase at Ser428 provides a potential para-
digm for the metabolic control of chloroplast carbon
metabolism. Biochem J 458:313–322
Satoh A, Kurano N, Harayama S, Miyachi S (2004) Effects of
chloramphenicol on photosynthesis, protein profiles and
transketolase activity under extremely high CO2 concen-
tration in an extremely-high-CO2-tolerant green microalga,
Chlorococcum littorale. Plant Cell Physiol 45:1857–1862
Schmitz K, Srivastava LM (1979) Long distance transport in
Macrocystis integrifolia. I. Translocation of 14C-labeled
assimilates. Plant Physiol 63:995–1002
123
Phytochem Rev
Schnarrenberger C, Flechner A, Martin W (1995) Enzymatic
evidence for a complete oxidative pentose-phosphate
pathway in chloroplasts and an incomplete pathway in the
cytosol of spinach leaves. Plant Physiol 108:609–614
Scruel O, Vanhoutte C, Sener A, Malaisse WJ (1998) Interfer-
ence of D-mannoheptulose with D-glucose phosphorylation,
metabolism and functional effects: comparison between
liver, parotid cells and pancreatic islets. Mol Cell Biochem
187:113–120
Sephton HH, Richtmyer NK (1963) The isolation of a second
octulose and of a heptose from the avocado: D-glycero-L-
galacto-octulose and D-glycero-D-galacto-heptose. J Org
Chem 28:1691–1694
Sephton HH, Richtmyer NK (1966) The isolation of D-erythro-
D-galacto-nonulose from the avocado, together with its
synthesis and proof of structure through reduction to D-
arabino-D-manno-nonitol and D-arabino-D-gluco-nonitol.
Carbohydr Res 2:289–300
Shaw PE, Wilson CW III, Knight RJ Jr (1980) High-perfor-
mance liquid chromatographic analysis of D-manno-hep-
tulose, perseitol, glucose, and fructose in avocado
cultivars. J Agric Food Chem 28:379–382
Shinmachi F, Buchner P, Stroud JL, Parmar S, Zhao FJ,
McGrath SP, Hawkesford MJ (2010) Influence of sulfur
deficiency on the expression of specific sulfate transporters
and the distribution of sulfur, selenium, and molybdenum
in wheat. Plant Physiol 153:327–336
Smeekens S, Ma J, Hanson J, Rolland F (2010) Sugar signals
and molecular networks controlling plant growth. Curr
Opin Plant Biol 13:274–279
Soria AC, Sanz ML, Villamiel M (2009) Determination of
minor carbohydrates in carrot (Daucus carota L.) by GC–
MS. Food Chem 114:758–762
Stankovič L, Kocková-Kratochvı́lová A, Bı́lik V (1986)
Isomerization of D-glycero-D-galacto-heptose to D-manno-
heptulose by selected yeasts. Folia Microbiol 31:344–345
The KEGG Database (2016) Kyoto Bioinformatics Centre,
Japan. http://www.genome.jp. Cited 18 January 2016
Takano J, Noguchi K, Yasumori M, Kobayashi M, Gajdos Z,
Miwa K, Hayashi H, Yoneyama T, Fujiwara T (2002)
Arabidopsis boron transporter for xylem loading. Nature
420:337–340
Takano J, Wada M, Ludewig U, Schaaf G, von Wirén N, Fuji-
wara T (2006) The Arabidopsis major intrinsic protein
NIP5;1 is essential for efficient boron uptake and plant
development under boron limitation. Plant Cell
18:1498–1509
Tesfay SZ, Bertling I, Bower JP (2012) D-Mannoheptulose and
perseitol in ‘Hass’ avocado: metabolism in seed and
mesocarp tissue. S Afr J Bot 79:159–165
Tolbert NE, Nystrom CW, Kerr PC (1957) Sedoheptulose in
Coleus. Plant Physiol 32:269–274
Tullson PC, Aprille JR (1986) Increased adenine nucleotides in
liver mitochondria after mannoheptulose injection in vivo.
Arch Biochem Biophys 246:611–616
Turgeon R, Wolf S (2009) Phloem transport: cellular pathways
and molecular trafficking. Ann Rev Plant Biol 60:207–221
Uematsu K, Suzuki N, Iwamae T, Inui M, Yukawa H (2012)
Increased fructose 1,6-bisphosphate aldolase in plastids
enhances growth and photosynthesis of tobacco plants.
J Exp Bot 63:3001–3009
Phytochem Rev
Valvano MA, Messner P, Kosma P (2002) Novel pathways for
biosynthesis of nucleotide-activated glycero-manno-hep-
tose precursors of bacterial glycoproteins and cell surface
polysaccharides. Microbiology 148:1979–1989
Villafranca JJ, Axelrod B (1971) Heptulose synthesis from
nonphosphorylated aldoses and ketoses by spinach trans-
ketolase. J Biol Chem 246:3126–3131
Whiley AW, Schaffer B, Lara SP (1991) Carbon dioxide
exchange of developing avocado (Persea americana Mill.)
fruit. Tree Physiol 11:85–94
Williams JF, MacLeod JK (2006) The metabolic significance of
octulose phosphates in the photosynthetic carbon reduction
cycle in spinach. Photosynth Res 90:125–148
Williams JF, Blackmore PF, Clark MG (1978) New reaction
sequences for the non-oxidative pentose phosphate path-
way. Biochem J 176:257–282
Wöber G, Hoffmann-Ostenhof O (1970a) Studies on the
biosynthesis of cyclitols, XXVI. Formation of mytilitol (C-
methyl-scyllo-inositol) in Chlorella fusca. Monatsh Chem
101:1861–1863
Wöber G, Hoffmann-Ostenhof O (1970b) Studies on the
biosynthesis of cyclitols. 28. On the biosynthesis of
laminitol (D-4-C-methyl-myo-inositol) in Porphyridium sp.
Eur J Biochem 17:393–396
Woloszczuk W, Hoffmann-Ostenhof O (1974) Studies on the
biosynthesis of cyclitols, XXXI. Further investigations on
the biosynthesis of laminitol (1D-4-C-methyl-myo-inositol)
in Porphyridium sp. Hoppe Seylers Z Physiol Chem
355:633–639
Yadav UP (2014) The sucrose–trehalose 6-phosphate (Tre6P)
nexus: specificity and mechanisms of sucrose signalling by
Tre6P. J Exp Bot 65:1051–1068
Ye Y, Zhang L, Yang R, Luo Q, Chen H, Yan X, Tang H (2013)
Metabolic phenotypes associated with high-temperature
tolerance of Porphyra haitanensis strains. J Agric Food
Chem 61:8356–8363
Young M (1972) Seven-carbon sugars and alcohols in the algae:
abstracts of papers read at the winter meeting at Gold-
smiths’ College, London. Br Phycol J 7:279–285
123