Journal of the Science of Food and Agriculture J Sci Food Agric 88:248255 (2008)

Distribution of fatty acids in edible

organs and seed fractions of borage
(Borago ofcinalis L.)
Mercedes del Ro-Celestino,
1
Rafael Font
1
and Antonio de Haro-Bail on
2
1
IFAPA-Centro Alameda del Obispo (Junta de Andaluca), Apartado 3092, E-14080 C ordoba, Spain
2
Instituto de Agricultura Sostenible (CSIC), Apartado 4084, E-14080 C ordoba, Spain
Abstract
BACKGROUND: Borage (Borago ofcinalis L.) is currently used as a vegetable in the north of Spain. The edible
parts of the plant are the petioles, leaves and stems. The objective of this study was to determine the oil and fatty
acids content in the edible tissues (leaves, petioles and stems) and seed fractions (endosperm + cotyledon, seed
coat and elaiosome) of white- and blue-owered borage accessions.
RESULTS: Leaves showed higher mean values of oil content (2.7%) than petioles and stems (1.1% and 1.4%,
respectively) in both, blue- and white-owered material. The most abundant fatty acids in leaves were palmitic,
linoleic, -linolenic and stearidonic acids (about 75% of the total fatty acids), while in petioles and stems myristic
and linoleic acids were most abundant (about 60% of the total fatty acids). Palmitic, oleic, linoleic and -linolenic
(GLA) acids were the major fatty acids of seed coat and endosperm + cotyledon in borage seeds (85% of the total
fatty acids), while palmitic, stearic, oleic and linoleic acids were the most abundant fatty acids of elaisome in
borage seeds (80% of the total fatty acids).
CONCLUSION: This paper shows that green parts of borage contain substantial amounts of omega-3 and omega-6
fatty acids, which are essential fatty acids for animal and human nutrition. Thus, borage could be a power food
of the future because of its content of unsaturated fatty acids, particularly the essential fatty acids, which have
great potential to prevent cardiovascular disease, cancer and infectious diseases.
2007 Society of Chemical Industry
Keywords: Borago ofcinalis; essential fatty acids; leaf; stem and petiole lipids; seed fractions
INTRODUCTION
Borage (Borago ofcinalis L.) is a large plant with blue,
star-shaped owers found throughout Europe, North
Africa and North America. This species is cultivated in
Europe and the USA for the commercial production
of seeds.
Seeds of borage are used in pharmaceutical products
because they are rich in -linolenic (all-cis-6,9,12-
octadecatrienoic) acid, an essential and unusual fatty
acid. This fatty acid is an intermediate of indispensable
compounds in the body, such as prostaglandin E
1
and
its derivatives.
13
Although B. ofcinalis seeds have been the objective
of various studies as a potential source of -linolenic
acid (GLA), no attention has been paid to date in
studying the oil content and fatty acid composition of
each seed fraction (endosperm + cotyledon, seed coat
and elaiosome), being the endosperm+ cotyledon the
part of the seed used for pharmaceutical purposes.
47
Information about the distribution of mass, oil and
fatty acid composition of the different seed parts could
be used in practical breeding programs in order to
increase oil content of the seed by means of the
combination of genotypes carrying genes with reduced
seed coat thickness and also genotypes with high oil
content, as occurs in other oleaginous species.
8,9
Borage plants have been used from ancient times as
a vegetable as well as in traditional medicine for the
treatment of swelling and inammation, coughs and
other respiratory complaints.
10,11
Nowadays, white-
owered borage is currently used as a vegetable in
the north of Spain, mainly in the Ebro Valley, where
the petioles, leaves and stems are highly appreciated;
they are eaten raw in salads or fried lightly, without
affecting the nutritional attributes of the plant. The
amount of borage plants consumed in the diet of
people in northern Spain is approximately 18 656t.
12
At the early stage of plant growth, 60% of the plant
matter is composed of leaves and the rest (40%) of
petioles. If the plant is harvested at later stages then
the stem represents about 10% of the whole plant.
The leaves of borage plants contain -linolenic
and stearidonic acids as major constituents of the
photosynthetic tissues, but low GLA content.
13

Correspondence to: Mercedes del Ro-Celestino, IFAPA-Centro Alameda del Obispo (Junta de Andaluca), Apartado 3092, E-14080 C ordoba, Spain
E-mail: mdrc@cica.es
(Received 11 April 2007; revised version received 8 July 2007; accepted 13 July 2007)
Published online 19 September 2007; DOI: 10.1002/jsfa.3080
2007 Society of Chemical Industry. J Sci Food Agric 00225142/2007/$30.00
Distribution of fatty acids in edible organs and seed fractions of borage
-Linolenic acid (18:3 n-3) plays an important
role in human growth, development and disease
prevention, being a precursor of the longer-chain
omega-3 fatty acids, eicosapentanoic acid (EPA),
docosapentaenoic acid (DPA) and docosahexaenoic
acid (DHA). Stearidonic acid (18:4 n-3) is also
important as a precursor in the metabolic pathway
leading to the formation of longer-chain fatty acids
and prostaglandin and therefore plays a key role in
human metabolism.
Although determination of the distribution of fatty
acids has been attempted in the leaf of this species,
until now it has not been studied in stems, petioles or
in the various fractions of borage seed. In this study,
the fatty acid prole of several accessions (cultivated
and wild) in borage edible parts and the fatty acid
composition within each fraction of borage seed were
analysed by chromatographic methods.
MATERIALS AND METHODS
Material
The germplasm evaluated consisted of accessions of
white- and blue-owered material and two types of
mutants of borage anatomically and cytogenetically
characterized in previous studies.
14,15
Four accessions
were chosen to conduct this work because of their
good agronomic behaviour in the semi-arid conditions
of Andalusia. In addition, the mutant lines were chosen
for their improved agronomic characteristics, one for
increased seed production per ower (Type B) and
the other for retaining mature seeds until harvest
(Type C mutant). The material was composed of the
following lines: (i) white-owered accession cultivated
as a vegetable in the north of Spain (southern
Europe) and provided by the Centre of Agricultural
Research of Rioja for this study; (ii) accession of blue-
owered roadside populations collected in southern
Spain for convenience, hereafter in this paper the
two accessions of cultivated and wild borage are
called WFA (white-owered accession) and BFA
(blue-owered accession), respectively; (iii) mutant
Type B: plants with modied owers larger than
normal, with a higher number of petals, sepals and
ovules (almost double that in the original plants);
(iv) mutant Type C: plants with owers closed.
These mutants were obtained in the course of a
mutagenesis programme with ethyl methanesulfonate
(EMS). In November 1993, approximately 15 000
seeds of the line RG-001 (white-owered borage
with high self-fertility) were exposed to a solution
of 1% (vol/vol) EMS for 16h under continuous
stirring. Plants from the M1 generation were bagged
to force self-pollination and mutants were identied
in the M2 generation of plants.
14
Seeds of each
accession of borage are deposited at the Herbarium
of the Department of Agronomy and Plant Breeding,
Instituto de Agricultura Sostenible (CSIC, C ordoba,
Spain).
Ten plants of each of the accessions tested in this
study were grown under greenhouse conditions at
20/15

C (day/night) with an 18h day length. Samples

for oil and fatty acid analyses were collected at 45
(petioles and leaves of rosette) and 60days (stems)
after planting, which is the optimal growth time for
consuming borage.
For analysing seed fractions, 10 seeds were
randomly selected and analysed for each of the
accessions used in this work. Seed coat, endosperm
+ cotyledons and elaiosome were separated manually
by cutting each of the seeds with a ne sharp blade
(Fig. 1).The seed coat, endosperm + cotyledons and
Figure 1. Seed fractions of borage.
J Sci Food Agric 88:248255 (2008) 249
DOI: 10.1002/jsfa
M del Ro-Celestino, R Font, A de Haro-Bail on
elaisome were dried and ground to a particle size of
<0.5mmbefore chemical analysis. The proportions of
seed coat, endosperm + cotyledons and elaiosome in
the different accessions were determined by weighing
100 seeds, in which each value was an average of three
determinations and expressed as individual weight
percentages per 100 seeds.
Methods
Fatty acid composition of the seed oil was determined
by gasliquid chromatography of fatty acid methyl
esters. Triheptadecanoic acid glycerate (T17:0) (from
Sigma, St Louis, MO, USA) was used as an internal
standard in order to quantify simultaneously the oil
content. Digestion, transmethylation of lipids from all
accessions (petioles, leaves, stems and seed fractions
of borage) and extraction of fatty acid methyl esters
in one step were performed using a PerkinElmer
Autosystem gasliquid chromatograph (PerkinElmer
Corporation, Norwalk, CT, USA) equipped with a
ame ionization detector (FID) and split injector.
16
The chromatograph was equipped with a capillary
column (25m0.25mm, i.d. 0.25m lm) with
acidied polyethylene glycol as the stationary phase.
Oven temperature was programmed from 195 to
225

C at a rate of 2

C min
1
. The temperature
of the detector and injector were 275 and 250

C,
respectively. Nitrogen was used as carrier gas. Fatty
acids were identied by comparing the retention times
of the borage methyl esters with those of known
mixtures of standard fatty acids (Sigma) run on the
same column under the same conditions.
Statistical analysis
One-way ANOVA calculation was carried out to
compare the mean values for essential fatty acids in
different edible parts of the plant and seed fractions
of borage, followed by Duncans multiple range test.
In addition, a statistical correlation among the seed
fractions of borage was performed.
RESULTS AND DISCUSSION
Organs of borage plant
Table 1 shows the oil content and fatty acid
composition in the edible portion of the different
sources of borage evaluated in this work. The Type
B mutant of borage showed values for oil and fatty
acid composition not signicantly different from the
Type C mutant, and both mutants showed oil and
fatty acid content that were not signicantly different
from those of the WFA, so that data for all these plants
were pooled for analysis.
Leaves showed signicantly higher mean values of
oil content (2.7%) than petioles and stems in both
BFA and WF material. Leaf lipids contained palmitic
(16:0), linoleic (18:2 n-6), -linolenic (18:3 n-3) and
stearidonic (18:4 n-3) acids as the major constituents,
which together composed about the 75% in BF
accession and 80% in WF material of the total fatty
acids. A comparison of the proportions of GLA and
stearidonic acids with those found in other members
of the family Boraginaceae shows that borage leaves
are one of the best sources of stearidonic acid. The
leaves of borage, however, are a moderate source of
GLA, being less productive than the leaves of Myosotis
scorpioides and Myosotis arvensis.
17
The most abundant fatty acids of petiole and stem
lipids were myristic (14:0) and linoleic (18:2 n-6)
acids, which represent about 60% in BFA and 50%
in WFA, respectively, of the total fatty acids. Petiole
and stem lipids of BFA contained about 4557% and
1318% of myristic and linoleic acids, respectively.
Petiole and stem lipids of WF material presented
2527% and 2023% of myristic and linoleic acids,
respectively.
The saturated fatty acids myristic and palmitic
showed high content in all the organs of the plants.
Petiole lipids showed the highest mean content of
myristic acid: 56.6% and 27.7% in BFA and WF
material, respectively. This nding is not in agreement
with the results found by other authors, in which
myristic acid is not present in lipids of rosette and
stalk leaves analysed during the growth cycle of the
common borage plant.
18
However, the results are in
agreement with those reported by Peiretty et al., who
showed the presence of myristic acid and variations in
the content of this fatty acid in ve stages of maturity
in common borage plants from late vegetative to early
seed stage, reaching the highest content at the early
owering stage.
19
Leaf lipids showed the highest mean content of
palmitic acid, with 19.8% and 24.4% in BFA and WF
material, respectively. Stearic (18:0) and oleic (18:1
n-9) acids showed lowvalues in all the tissues analysed.
The percentage of linoleic acid was high in all the
edible portions of the borage plant, with a range of
variation from 9.8% in leaves to 23.0% in stems, both
of WF material.
GLA was present in all the organs analysed and
showed signicant differences between different parts
of the plant in both BFA and WF material. The
content of GLA was highest in stems, ranging from
10.2% to 11.5%, with an average of 11.0% and 10.3%
in BFA and WFA, respectively. Leaves showed the
lowest mean content of GLA, ranging from 2.1% to
6.5% with mean values of 5.0% and 3.3% in BFA and
WFA, respectively.
All the organs of borage plants investigated had
GLA and stearidonic acid, in an approximate ratio
of 1:3, 2:1 and 3:1 for leaves, petioles and stems,
respectively. Leaves showed the highest values of -
linolenic acid in both BFA and WF material, with
26.2% and 32.2%, respectively. Stearidonic acid was
synthesized in substantial amounts in all the organs
evaluated, with the highest levels in leaves (15.2%).
The presence of -linolenic, stearidonic acids and
GLA in all the tissues of borage indicates that both
-15-and -6-desaturases were active.
250 J Sci Food Agric 88:248255 (2008)
DOI: 10.1002/jsfa
Distribution of fatty acids in edible organs and seed fractions of borage
T
a
b
l
e
1
.
M
e
a
n
a
n
d
r
a
n
g
e
(
n
=
1
0
)
f
o
r
o
i
l
c
o
n
t
e
n
t
a
n
d
f
a
t
t
y
a
c
i
d
c
o
m
p
o
s
i
t
i
o
n
o
f
l
e
a
f
,
p
e
t
i
o
l
e
a
n
d
s
t
e
m
l
i
p
i
d
s
o
f
d
i
f
f
e
r
e
n
t
b
o
r
a
g
e
a
c
c
e
s
s
i
o
n
s
(
B
F
A
,
b
l
u
e
-

o
w
e
r
e
d
a
c
c
e
s
s
i
o
n
s
;
W
F
A
,
w
h
i
t
e
-

o
w
e
r
e
d
a
c
c
e
s
s
i
o
n
s
)
F
a
t
t
y
a
c
i
d
c
o
m
p
o
s
i
t
i
o
n
(
%
o
f
t
o
t
a
l
f
a
t
t
y
a
c
i
d
s
i
n
t
h
e
o
i
l
)
O
i
l
(
%
)
1
4
:
0
0
1
6
:
0
0
1
8
:
0
1
8
:
1
n
-
9
1
8
:
2
n
-
6
1
8
:
3
n
-
6
1
8
:
3
n
-
3
1
8
:
4
n
-
3
2
2
:
1
n
-
9
O
t
h
e
r
s
a
L
e
a
v
e
s
B
F
A
2
.
7
1
1
.
8
1
9
.
8
3
.
0
1
.
9
1
2
.
3
5
.
0
2
6
.
2
1
5
.
2
0
.
6
2
.
6
1
.
9

3
.
4
7
.
8

1
5
.
6
1
6
.
1

2
3
.
5
2
.
9

3
.
2
1
.
3

2
.
9
1
0
.
4

1
4
.
3
3
.
9

6
.
5
2
4
.
5

2
7
.
4
1
2
.
7

1
7
.
5
0
.
4

0
.
9
1
.
9

3
.
0
L
e
a
v
e
s
W
F
A
2
.
4
5
.
5
2
4
.
4
3
.
0
3
.
0
9
.
8
3
.
3
3
2
.
2
1
3
.
8
0
.
4
3
.
6
2
.
3

2
.
5
4
.
4

7
.
0
1
8
.
0

2
9
.
1
2
.
3

3
.
7
2
.
4

3
.
5
7
.
7

1
1
.
9
2
.
1

4
.
5
2
5
.
2

3
9
.
9
1
3
.
3

1
4
.
5
0
.
2

0
.
7
1
.
7

5
.
2
P
e
t
i
o
l
e
s
B
F
A
1
.
1
5
6
.
6
9
.
8
1
.
2
1
.
1
1
2
.
7
6
.
1
5
.
6
2
.
8
0
.
5
2
.
1
0
.
8

1
.
4
4
8
.
5

6
6
.
6
7
.
2

1
1
.
4
0
.
8

1
.
6
0
.
9

1
.
4
1
0
.
1

1
4
.
9
5
.
1

7
.
2
3
.
7

6
.
7
1
.
9

3
.
5
0
.
3

0
.
7
1
.
7

2
.
8
P
e
t
i
o
l
e
s
W
F
A
1
.
2
2
7
.
7
1
6
.
3
2
.
4
2
.
7
1
9
.
3
9
.
1
1
1
.
8
4
.
9
0
.
4
4
.
9
1
.
1

1
.
2
1
9
.
9

3
2
.
7
1
5
.
6

1
6
.
9
1
.
7

3
.
7
2
.
3

3
.
1
1
6
.
2

2
1
.
7
6
.
4

1
0
.
8
8
.
6

1
7
.
8
4
.
0

6
.
7
0
.
1

0
.
7
2
.
3

1
0
.
3
S
t
e
m
s
B
F
A
1
.
5
4
5
.
0
1
2
.
3
1
.
3
1
.
8
1
8
.
2
1
1
.
0
4
.
0
2
.
8
0
.
3
2
.
3
1
.
3

1
.
7
4
3
.
4

4
6
.
6
1
2
.
0

1
2
.
7
1
.
3

1
.
3
1
.
8

1
.
9
1
7
.
7

1
8
.
7
1
0
.
4

1
1
.
5
3
.
7

4
.
3
2
.
5

3
.
0
0
.
3

0
.
3
2
.
2

2
.
5
S
t
e
m
s
W
F
A
1
.
4
2
5
.
6
1
6
.
9
2
.
8
3
.
4
2
3
.
0
1
0
.
3
8
.
4
4
.
1
0
.
3
5
.
1
1
.
3

1
.
5
2
4
.
0

2
7
.
3
1
6
.
8

1
7
.
0
2
.
6

2
.
8
3
.
4

3
.
5
2
2
.
8

2
3
.
2
1
0
.
2

1
0
.
4
8
.
1

8
.
8
3
.
8

4
.
4
0
.
3

0
.
3
4
.
7

5
.
5
a

O
t
h
e
r
s

i
n
c
l
u
d
e
t
h
e
f
a
t
t
y
a
c
i
d
s
1
2
:
0
(
l
a
u
r
i
c
)
,
1
6
:
1
(
p
a
l
m
i
t
o
l
e
i
c
)
,
2
0
:
1
n
-
9
(
c
i
s
-
1
1
-
e
i
c
o
s
e
n
o
i
c
)
,
2
2
:
0
(
b
e
h
e
n
i
c
)
,
2
4
:
0
(
l
i
g
n
o
c
e
r
i
c
)
,
2
4
:
1
n
-
9
(
n
e
r
v
o
n
i
c
)
.
Erucic (22:1 n-9) acid content was present in all
the organs of borage plant in very low concentration
(0.30.9%).
Seeds
Table 2 shows the weight of 100 seeds and percentages
of seed coat, endosperm + cotyledon and elaiosome
of different seeds of borage. The average seed weight
of different accessions varied between 2.1 and 2.8 g
per 100 seeds. The weights of BFA (2.9g per100
seeds) were signicantly (P < 0.05) higher than that
of WFA (2.5 g per 100 seeds), Type B mutant (2.1 g
per 100 seeds) and Type C mutant (2.1 g per 100
seeds). The percentages of endosperm + cotyledon
and seed coat were the predominant fractions for all
accessions, with values of approximately 50% and
44%, respectively. The proportion of elaiosome was
a very small part of the seed and varied between
5.3% in Type B mutant and 7.2% in Type C mutant.
The present study showed a high negative correlation
between the seed coat and endosperm + cotyledon
of all the accessions of borage (r = 0.72, P < 0.01).
This value could be of interest in breeding for high oil
content in seeds if those accessions showing high oil
content are combined with genotypes carrying genes
with reduced seed coat content.
There were no signicant differences (P > 0.05) for
the percentages of different seed fractions among the
four accessions (Table 2).
The data given in Table 3 show the oil and fatty
acids content in the different parts of the borage seed.
The oil content in the different fractions of the borage
seed exhibited a wide variability, ranging from 1.0%
to 3.4% in elaiosome, from 1.8% to 3.9% in seed coat
and from 46.8% to 49.9% in endosperm+ cotyledon.
The fatty acid composition showed variation with
respect to specic parts of the borage seed, the pattern
being similar for seed coat and endosperm+cotyledon
and different for the elaiosome. Palmitic, oleic and
linoleic acids and GLA were the major fatty acids of
seed coat and endosperm+cotyledon in borage seeds,
which together composed about 85% of the total fatty
acids. The most abundant fatty acids of elaisome in
borage seeds were palmitic, stearic, oleic and linoleic
acids, which together composed about 80% of the
total fatty acids.
Table 2. Mean weight of 100 seeds (g/100 seeds) and percentages of
each seed fraction (seed coat, endosperm + cotyledon and
elaiosome) of different accessions of borage (WFA, white-owered
accession; BFA, blue-owered accession)
Accessions
Weight
100 seeds Elaiosome
Seed
coat
Endosperm +
cotyledon
WFA 2.5b 5.7a 44.4a 49.9a
BFA 2.9a 6.4a 42.8a 50.8a
Type B mutant 2.1b 5.3a 45.2a 49.4a
Type C mutant 2.1b 7.2a 44.1a 48.7a
Values followed by different letters within each column denote a
signicant difference and those followed by same letters denote no
signicant difference at P < 0.05.
J Sci Food Agric 88:248255 (2008) 251
DOI: 10.1002/jsfa
M del Ro-Celestino, R Font, A de Haro-Bail on
Elaiosome exhibited the highest palmitic and stearic
acid content, with maxima of 41.4% (Type B mutant)
and 14.9% (Type C mutant), respectively. Palmitic
acid content of elaiosome varied widely between
accessions, reaching a maximumof 42.1%in the Type
Bmutant of borage, and a minimumof 22.4%for Type
C mutant.
Percentage of GLA ranged from 0.4% in elaiosome
to 17% in endosperm + cotyledon. Traces of
GLA (0.40.7%) were present in elaiosome for all
accessions analysed, while the GLA content of seed
coat and endosperm + cotyledon ranged from 12.9%
to 16.9%. The values of GLA agreed with those
reported by Del Ro et al.
6
in evaluation of Spanish
borage populations; in that study an important range of
variation from 8.7% to 28.6% was observed, but these
gures were signicantly lower than those reported by
Janik et al.
5
and Galvey and Shirlin.
20
The percentages of -linolenic acid in total fatty
acids ranged from absence in endosperm + cotyledon
to 3.2% in elaiosome of BFA.
Stearidonic acid was present in the seed coat and
elaiosome of borage seeds, which exhibited levels that
ranged from 0.6% to 0.8% and from 1.5% to 4.1%,
respectively, but this fatty acid was not detected in
the endosperm + cotyledon fraction of the seed. It
seems likely that leaves, petioles and stems may share
common mechanisms for -linolenic and stearidonic
acid formation which differ from those of the storage
lipids of seeds. Different desaturation systems (or their
control) may predominate in photosynthetic versus
non-photosynthetic plant tissues. However, whether
the photosynthetic activity of a tissue plays a role in
the differential regulation among the various tissues
remains to be elucidated. The observations reported
here are consistent with previous studies of leaf lipids
in borage, which reported that stearidonic acid was
synthesized within the developing chloroplast in the
-15- and and -6-desaturases acting on linoleic
acid esteried to galactolipids.
13
Also, this nding
is in agreement with those reported for other species
such as soybean and evening primrose, in which seeds
may share common desaturation mechanisms for -
linolenic acid formation with roots, but differ from
those of leaves and stem.
21,22
With respect to cis-13-docosenoic (20:1 n-9) and
erucic acid content, both were present in all the parts
of the seed and their concentrations were very similar,
ranging from 1% to 4.4% for cis-13-docosenoic acid
and from 1.7% to 3.4% for erucic acid, in an
approximate ratio of 1:1.
Total unsaturated and essential fatty acids
tissues of plant and seed fractions
Signicant differences existed in total fatty acid
contents between different plant tissues and seed
fractions (Figs 2 and 3). On a dry mass basis, the
highest contents of total fatty acids (essential and
other fatty acids) were in the seeds, followed by
the leaves and stems, while the lowest were found
in petioles (Figs 2 and 3). Taking into account that
seeds are the storage organs is not surprising to nd
higher levels of total fatty acids than in green parts
of the plant in order to assist their dispersal and to
provide an efcient source of energy for the growth of
new seedlings. The total fatty acid content in leaves
ranged from 2.4% to 2.7%, in stems from 1.4% to
1.5%, and in petioles from 1.1% to 1.2%. The highest
total content of unsaturated fatty acids was reached
in leaves of BFA with 17.1mg g
1
dry mass, then
in stems of WFA with 6.3mg g
1
and in petioles
of WFA with 5.5mg g
1
. Based on the analysis of
total content of unsaturated fatty acids in borage
tissues, the levels detected were slightly lower than
those of other varieties with great potential as a green
leafy vegetable (purslane, Portulaca oleracea L.) which
showed unsaturated fatty acid contents of 18.8mg g
1
in leaves and 12mg g
1
in stems.
23
The total essential fatty acid content in borage plants
was 39.2% in stems, 44.8% in petioles and 60.0% in
BF WF BF WF BF WF
0
5
10
15
20
25
30
b
a
a
a
a
a
m
g

g

1

d
r
y

m
a
s
s
total essential fatty acids
other fatty acids
Leaves Petioles Stems
Figure 2. Total fatty acids (essential and other fatty acids) relative to dry mass of leaves, petioles and stems of different borage accessions. For
total essential fatty acids and part of the plant, values followed by different letters denote a signicant difference, while those followed by the same
letter denote no signicant difference at P < 0.05.
252 J Sci Food Agric 88:248255 (2008)
DOI: 10.1002/jsfa
Distribution of fatty acids in edible organs and seed fractions of borage
T
a
b
l
e
3
.
M
e
a
n
a
n
d
r
a
n
g
e
(
n
=
1
0
)
f
o
r
o
i
l
c
o
n
t
e
n
t
a
n
d
f
a
t
t
y
a
c
i
d
c
o
m
p
o
s
i
t
i
o
n
i
n
e
a
c
h
s
e
e
d
f
r
a
c
t
i
o
n
(
s
e
e
d
c
o
a
t
,
e
n
d
o
s
p
e
r
m
+
c
o
t
y
l
e
d
o
n
a
n
d
e
l
a
i
o
s
o
m
e
)
o
f
d
i
f
f
e
r
e
n
t
b
o
r
a
g
e
a
c
c
e
s
s
i
o
n
s
(
W
F
A
,
w
h
i
t
e
-

o
w
e
r
e
d
a
c
c
e
s
s
i
o
n
;
B
F
A
,
b
l
u
e
-

o
w
e
r
e
d
a
c
c
e
s
s
i
o
n
)
F
a
t
t
y
a
c
i
d
c
o
m
p
o
s
i
t
i
o
n
(
%
o
f
t
h
e
t
o
t
a
l
f
a
t
t
y
a
c
i
d
s
i
n
t
h
e
o
i
l
)
O
i
l
c
o
n
t
e
n
t
(
%
d
.
m
.
)
1
4
:
0
0
1
6
:
0
0
1
8
:
0
1
8
:
1
n
-
9
1
8
:
2
n
-
6
1
8
:
3
n
-
6
1
8
:
3
n
-
3
1
8
:
4
n
-
3
2
0
:
1
n
-
9
2
2
:
1
n
-
9
O
t
h
e
r
s
a
T
y
p
e
B
M
u
t
a
n
t
E
n
d
o
s
p
e
r
m
+
c
o
t
y
l
e
d
o
n
4
9
.
9
0
.
0
1
2
.
6
5
.
2
2
2
.
7
3
3
.
2
1
6
.
6
0
.
0
0
.
0
3
.
9
3
.
0
2
.
6
4
6
.
3

5
3
.
3
0
.
0

0
.
0
1
2
.
5

1
2
.
7
5
.
1

5
.
3
2
2
.
4

2
2
.
9
3
2
.
9

3
3
.
4
1
6
.
3

1
6
.
9
0
.
0

0
.
0
0
.
0

0
.
0
3
.
8

4
.
1
2
.
8

3
.
2
2
.
4

2
.
8
S
e
e
d
c
o
a
t
1
.
9
0
.
0
1
7
.
5
5
.
1
1
9
.
1
3
3
.
6
1
4
.
7
0
.
5
0
.
6
3
.
2
3
.
2
2
.
4
1
.
2

2
.
6
0
.
0

0
.
0
1
6
.
6

1
8
.
4
5
.
0

5
.
2
1
9
.
0

1
9
.
1
3
3
.
4

3
3
.
7
1
3
.
5

1
5
.
9
0
.
2

0
.
6
0
.
6

0
.
6
3
.
2

3
.
3
3
.
1

3
.
3
2
.
0

2
.
7
E
l
a
i
o
s
o
m
e
1
.
0
0
.
0
4
1
.
4
1
0
.
3
1
2
.
8
1
9
.
9
0
.
7
0
.
6
1
.
7
1
.
1
5
.
0
0
.
9
1
.
0

1
.
1
0
.
0

0
.
0
4
0
.
7

4
2
.
1
8
.
9

1
1
.
7
1
2
.
7

1
2
.
9
1
8
.
6

2
1
.
1
0
.
6

0
.
8
0
.
5

0
.
7
1
.
2

2
.
2
0
.
9

1
.
2
4
.
9

5
.
0
0
.
4

1
.
4
T
y
p
e
C
M
u
t
a
n
t
E
n
d
o
s
p
e
r
m
+
c
o
t
y
l
e
d
o
n
4
8
.
5
0
.
0
1
2
.
9
4
.
6
2
3
.
7
3
3
.
6
1
4
.
9
0
.
0
0
.
0
4
.
3
3
.
0
2
.
6
4
7
.
2

5
1
.
4
0
.
0

0
.
0
1
2
.
8

1
3
.
1
4
.
5

4
.
7
2
3
.
1

2
4
.
2
3
3
.
2

3
3
.
9
1
4
.
5

1
5
.
4
0
.
0

0
.
0
0
.
0

0
.
0
4
.
1

4
.
5
2
.
9

3
.
2
2
.
5

3
.
1
S
e
e
d
c
o
a
t
3
.
9
0
.
0
1
6
.
8
4
.
7
2
0
.
2
3
3
.
7
1
4
.
9
0
.
4
0
.
6
3
.
6
3
.
0
2
.
1
3
.
7

4
.
1
0
.
0

0
.
0
1
6
.
6

1
7
.
0
4
.
6

4
.
8
2
0
.
2

2
0
.
2
3
3
.
6

3
3
.
8
1
4
.
4

1
5
.
4
0
.
2

0
.
5
0
.
5

0
.
6
3
.
6

3
.
6
2
.
9

3
.
1
1
.
9

2
.
2
E
l
a
i
o
s
o
m
e
3
.
4
0
.
2
2
3
.
1
1
4
.
9
3
4
.
4
1
0
.
2
0
.
4
0
.
7
4
.
1
2
.
5
1
.
6
0
.
9
3
.
1

3
.
8
0
.
0

0
.
4
2
2
.
4

2
3
.
8
1
4
.
9

1
4
.
9
3
3
.
7

3
5
.
2
1
0
.
1

1
0
.
2
0
.
4

0
.
5
0
.
6

0
.
8
3
.
9

4
.
2
2
.
5

2
.
5
1
.
5

1
.
7
0
.
7

1
.
2
B
F
A
E
n
d
o
s
p
e
r
m
+
c
o
t
y
l
e
d
o
n
4
9
.
9
0
.
0
1
2
.
0
4
.
8
3
0
.
5
2
7
.
5
1
4
.
3
0
.
0
0
.
0
4
.
3
3
.
2
3
.
0
4
3
.
6

5
7
.
7
0
.
0

0
.
0
1
1
.
5

1
2
.
4
4
.
4

5
.
0
2
8
.
1

3
3
.
5
2
6
.
4

2
8
.
9
1
2
.
8

1
5
.
9
0
.
0

0
.
0
0
.
0

0
.
0
4
.
1

4
.
6
3
.
0

3
.
5
2
.
7

3
.
3
S
e
e
d
c
o
a
t
1
.
8
0
.
0
1
7
.
6
4
.
6
2
6
.
7
2
7
.
3
1
2
.
6
0
.
7
0
.
8
3
.
5
3
.
3
2
.
2
1
.
5

2
.
1
0
.
0

0
.
0
1
6
.
5

1
9
.
0
4
.
2

5
.
1
2
5
.
1

3
0
.
3
2
5
.
8

2
8
.
7
1
1
.
5

1
3
.
9
0
.
5

0
.
8
0
.
7

0
.
9
3
.
3

3
.
7
3
.
1

3
.
4
2
.
1

2
.
3
E
l
a
i
o
s
o
m
e
2
.
0
1
.
9
3
1
.
0
7
.
9
2
7
.
4
1
5
.
6
0
.
4
3
.
2
1
.
5
0
.
9
1
.
7
3
.
9
1
.
4

2
.
8
1
.
0

3
.
1
2
8
.
8

3
3
.
5
6
.
9

8
.
9
2
5
.
6

2
8
.
7
1
4
.
0

1
6
.
4
0
.
4

0
.
5
2
.
8

3
.
6
1
.
0

2
.
1
0
.
7

1
.
2
1
.
3

2
.
5
3
.
2

4
.
3
W
F
A
E
n
d
o
s
p
e
r
m
+
c
o
t
y
l
e
d
o
n
4
6
.
8
0
.
0
1
2
.
4
5
.
2
2
7
.
1
3
1
.
0
1
3
.
9
0
.
0
0
.
0
4
.
4
3
.
1
2
.
6
4
1
.
1

5
2
.
7
0
.
0

0
.
0
1
2
.
2

1
2
.
5
5
.
1

5
.
3
2
6
.
2

2
8
.
2
3
0
.
6

3
1
.
4
1
2
.
9

1
4
.
7
0
.
0

0
.
0
0
.
0

0
.
0
4
.
3

4
.
6
3
.
0

3
.
2
2
.
5

2
.
8
S
e
e
d
c
o
a
t
1
.
8
0
.
2
1
8
.
2
4
.
7
2
1
.
2
3
2
.
2
1
2
.
7
0
.
8
0
.
7
3
.
3
3
.
1
2
.
3
1
.
7

1
.
8
0
.
0

0
.
4
1
7
.
8

1
8
.
5
4
.
7

4
.
8
2
0
.
7

2
1
.
6
3
1
.
9

3
2
.
4
1
2
.
6

1
2
.
7
0
.
6

0
.
9
0
.
7

0
.
7
3
.
3

3
.
4
3
.
1

3
.
1
2
.
1

2
.
6
E
l
a
i
o
s
o
m
e
2
.
7
0
.
4
2
9
.
3
1
0
.
4
2
8
.
7
1
2
.
8
0
.
4
0
.
3
3
.
5
4
.
0
2
.
2
1
.
7
2
.
5

3
.
0
0
.
0

0
.
9
2
8
.
7

2
9
.
8
1
0
.
1

1
0
.
8
2
7
.
3

3
0
.
1
1
2
.
2

1
3
.
4
0
.
4

0
.
4
0
.
3

0
.
4
3
.
3

3
.
8
3
.
7

4
.
2
1
.
8

2
.
5
0
.
9

2
.
4
a

O
t
h
e
r
s

i
n
c
l
u
d
e
t
h
e
f
a
t
t
y
a
c
i
d
s
1
2
:
0
(
l
a
u
r
i
c
)
,
1
6
:
1
(
p
a
l
m
i
t
o
l
e
i
c
)
,
2
0
:
1
n
-
9
(
c
i
s
-
1
1
-
e
i
c
o
s
e
n
o
i
c
)
,
2
2
:
0
(
b
e
h
e
n
i
c
)
,
2
4
:
0
(
l
i
g
n
o
c
e
r
i
c
)
a
n
d
2
4
:
1
n
-
9
(
n
e
r
v
o
n
i
c
)
.
J Sci Food Agric 88:248255 (2008) 253
DOI: 10.1002/jsfa
M del Ro-Celestino, R Font, A de Haro-Bail on
Endosperm + cotyledon Seed coat Elaiosome
B
F
W
F
T
y
p
e

B
T
y
p
e

C
B
F
W
F
T
y
p
e

B
T
y
p
e
C
B
F
W
F
T
y
p
e

B
T
y
p
e

C
0
100
200
300
400
500
b b
a
b bc
b
b
a
a
a
a
a
m
g

g

1

d
r
y

m
a
s
s
total essential fatty acids
other fatty acids
Figure 3. Total fatty acids (essential and other fatty acids) relative to dry mass of endosperm + cotyledons, seed coat and elaiosome of different
borage accessions. For total essential fatty acids and fraction of seed, values followed by different letters denote a signicant difference, while
those followed by the same letter denote no signicant difference at P < 0.05.
leaves. Signicant differences (P < 0.05) were only
found in total essential fatty acid contents between
accessions in petioles, the highest content being in
petioles of WFA (Fig. 1). Petioles of WFA showed
a higher content of the essential fatty acids linoleic,
GLA, -linolenic and stearidonic acids than those of
BFA (Table 1). These differences detected for total
essential fatty acid content between petioles of BF
and WF material suggest that other accessions of
borage could be evaluated to identify higher levels
than those found in this work. WF material were
part of a borage collection provided by the Centre
of Agricultural Research of Rioja (130 accessions),
which was evaluated in a previous study for variants
in oil content and fatty acid composition of the seed,
plant habit, maturity and seed production, exhibiting
a great range of variation for all the traits studied. It
is expected that further studies of the large available
Spanish borage collection will reveal additional useful
variation for total essential fatty acid in petioles.
6
The seed fractions that showed the highest total
unsaturated and essential fatty acids were the
endosperm + cotyledon, with 82.6% and 73% (412.4
and 364mg g
1
) in BFA, then seed coat, with
77.6% and 69.3% (30.1 and 26.8mg g
1
) in Type
C mutant, and elaisome with 54.3% and 49.8% (18.6
and 17.0mg g
1
) in Type C mutant, respectively.
Signicant differences (P < 0.05) existed in total
essential fatty acid contents between accessions by
different seed fractions. Type C mutant exhibited
signicant higher essential fatty acid content in seed
coat and elaiosome than other accessions (Fig. 2).
Both Type B and Type C mutants of borage were
isolated after chemical mutagen treatment of an
agronomically good line of WF borage (RG-001). In
previous studies of characterization and anatomical
studies in relation to these mutant lines it was
hypothesised that, on one hand, Type B mutant was
the result of quantitative changes in the borage genome
and, on the other hand, Type C mutant was the
result of localized mutations in the borage genome.
14
In Type C mutant the petals and petal accessory
structures remain present throughout fruit maturation,
pressing and retaining the seeds on the fruit receptacle.
Additionally, in this mutant line the elaiosome is
embedded in the fruit receptacle, contributing to seed
retention.
15
Our preliminary results in relation to the
differences observed in the oil and fatty acid content
in seed coat and especially in the elaisome of Type C
mutant (Table 3) could contribute to the retention of
mature seeds until harvest in these plants, eliminating
seed loss due to seed shattering that characterizes
normal plants.
The occurrence of high concentrations of total
unsaturated fatty acids in the endosperm + cotyledon
of borage seeds in our study is comparable to
that of Sesbania aculeate seeds (78.181.3%). These
values were higher than those of Sesbania bispinosa
(66.7%), cow pea (68.1%) and chick pea (67.1%),
and slightly lower than those of eld pea (85.0%),
beach pea (85.2%) and green pea (83.5%).
24,25
The
high content in unsaturated fatty acids (82.6%) and
lower proportion of saturated fatty acids (17.4%) in
the oil of borage seeds make it very appropriate for use
in the food industry. Additionally, the high amount of
GLA in this oil reinforces its nutritional value.
CONCLUSIONS
This study demonstrates that borage contains sub-
stantial amounts of omega-3 and omega-6 fatty acids
(linoleic, -linolenic, GLA and stearidonic acids),
which are essential for animal and human nutrition.
The levels detected in leaves, petioles, stems and seed
fractions were comparable to those of green leafy veg-
etables. The borage plant could be a power food
of the future because of its content of unsaturated
fatty acids, particularly the essential fatty acids, which
254 J Sci Food Agric 88:248255 (2008)
DOI: 10.1002/jsfa
Distribution of fatty acids in edible organs and seed fractions of borage
exhibit a great potential to prevent cardiovascular dis-
ease, cancer and infectious diseases.
New studies are being carried out in our research
group for determining other nutritional attributes such
as vitamins A, C and E and -carotenes.
ACKNOWLEDGMENTS
The authors wish to extend their appreciation to Dr
Vicente Dominguez (Centre of Agricultural Research
of Rioja) for providing the white-owered accession
of borage. We would like to thank Gloria Fern andez
Marn (Instituto de Agricultura Sostenible, CSIC,
C ordoba) for her technical assistance. M del Ro-
Celestino is nanced by the Ram on y Cajal program
(Ministerio de Ciencia y Tecnologa).
REFERENCES
1 Horrobin DF, The role of essential fatty acids and prostaglandins
in the premenstrual syndrome. J Reprod Med 28:465468
(1983).
2 Wolf RB, Kleiman R and England RE, New sources of gamma-
linolenic acid. J Am Oil Chem Soc 60:18581860 (1983).
3 Horrobin DF, Nutritional and medical importance of gamma-
linolenic acid. Prog Lipid Res 31:163194 (1992).
4 Whipkey A, Simon JE and Janick J, In vivo and in vitro lipid
accumulation in Borago ofcinalis L. J Am Oil Chem Soc
65:979984 (1988).
5 Janick J, Simon JE, Quinn J and Beaubaire N, Borage: a source
of gamma-linolenic acid, in Herbs, Spices and Medicinal Plants,
ed. by Cracker LE and Simon JE. Oryx Press, Phoenix, AZ,
pp. 210232 (1989).
6 Del Ro M, Fern andez-Martnez JM and De Haro A, Wild and
cultivated Borago ofcinalis L.: sources of gamma-linolenic
acid. Grasas y Aceites 44:125126 (1993).
7 De Haro A, Domnguez V and Del Ro M, Variability in the
content of gamma-linolenic acid and other fatty acids of the
seed oil of germplasm of wild and cultivated borage (Borago
ofcinalis L.). Journal of Herbs, Spices and Medicinal Plants
9:297304 (2002).
8 Fern andez-Martinez JM, Del Ro M and De Haro A, Survey of
safower (Carthamus tinctorius L.) germplasm for variants in
fatty acid composition and other seed characters. Euphytica
69:115122 (1993).
9 Miller JF and Fick GN, Sunower genetics. sunower tech-
nology and production, in Agronomy Monograph 35, ed.
by Schneiter AA. ASA, CSSA and SSSA, Madison, WI,
pp. 441495 (1997).
10 Gerard J, The History of Plants, ed. by Woodward M. Senate
Studio Editions, London, pp. 185 (1994).
11 Prakash V, Leafy Spices. CRC Press, Boca Raton, FL (1990).
12 Floris E, Montaner C, Borraja En Nuez FY and Llacer G, La
Horticultura Espa nola, ed. by Sociedad Espa nola de Ciencias
Hortcolas. Mundi-Prensa, Madrid, pp. 114116 (2001).
13 Grifths G, Brechany EY, Jackson FM, Christie WW, Stymne S
and Stobart AK, Distribution and biosynthesis of stearidonic
acid in leaves of Borago ofcinalis. Phytochemistry 43:381386
(1996).
14 De Haro A and Del Ro M, Isolation of chemically-induced
mutants in borage (Borago ofcinalis L.). J Am Oil Chem Soc
75:281283 (1998).
15 De Haro A, Del Ro M, Alcaide B, Rapoport H and Cabrera A,
Characterisation and evaluation of species of the Boraginaceae
family as source of gamma-linolenic acid for Mediterranean
conditions. Acta Hortic 629:231237 (2004).
16 Garces R and Mancha M, One-step lipid extraction and fatty
acid methyl esters preparation from fresh plant tissues. Anal
Biochem 211:139143 (1993).
17 Jamieson GR and Reid EH, The leaf lipids of some members of
the Boraginaceae family. Phytochemistry 8:14891494 (1969).
18 Sewon P and Tyystjarvi E, Stearidonic and gamma-linolenic
acid contents of common borage leaves. Phytochemistry
33:10291032 (1993).
19 Peiretti PG, Palmegiano GB and Salamano G, Quality and fatty
acid content of borage (Borago ofcinalis L.) during the growth
cycle. Ital J Food Sci 16:177184 (2004).
20 Galwey NW and Shirlin AJ, Selection of borage (Borago
ofcinalis) as a seed crop for pharmaceutical uses. Heredity
65:249257 (1990).
21 Wang X, Norman HA, St John JB, Yin T and Hildebrand DF,
Comparison of fatty acid composition in soybean tissues with
low linolenate mutants. Phytochemistry 28:411414 (1989).
22 Lotti G and Quartacci MF, La distribuzione dellacido gamma
linolenico nei fosfolipidi dei semi di Oenothera biennis L.
Agrochimica 34:2432 (1990).
23 Liu L, Howe P, Zhou YF, Xu ZQ, Hocart C and Zhang R,
Fatty acids and beta-carotene in Australian purslane
(Portulaca oleracea) varieties. J Chromatogr 893:207213
(2000).
24 Salunkhe DK, Sathe SK and Reddy NR, Legume lipids, in
Chemistry and Biochemistry of Legumes, ed. by Arona SK.
Oxford and LBH Publishing, New Delhi, pp. 51109 (1982).
25 Chavan UD, Shahidi F, Bal AK and Mckenzie DB, Physico-
chemical and nutrient composition of beach pea (Lathyrus
maritimus L.). Food Chem 66:4350 (1999).
J Sci Food Agric 88:248255 (2008) 255
DOI: 10.1002/jsfa