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Antifungal Activity of the Carrot Seed Oil and its Major Sesquiterpene Compounds

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Carrot seed oil is the source of the carotane sesquiterpenes carotol, daucol and-caryo-phyllene. These sesquiterpenic allelochemicals were evaluated against Alternaria alternata isolated from the surface of carrot seeds cultivar Perfekcja, a variety widely distributed in horticultural practise in Poland. Alternaria alternata is one of the most popular phytotoxic fungi infesting the carrot plant. The strongest antifungal activity was observed for the main constituent of carrot seed oil, carotol, which inhibited the radial growth of fungi by 65% at the following concentration.
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Antifungal Activity of the Carrot Seed Oil and its Major Sesquiterpene
Compounds
Izabela Jasicka-Misiak
a,*
, Jacek Lipok
a
, Ewa M. Nowakowska
a
, Piotr P. Wieczorek
a
,
Piotr Młynarz
b
, and Paweł Kafarski
a
a
Institute of Chemistry, University of Opole, Oleska 48, 45-052 Opole, Poland.
Fax: +4877 444401. E-mail: izajm@uni.opole.pl
b
Institute of Organic Chemistry, Biochemistry & Biotechnology, Wroclaw University of
Technology, Wybrzeze Wyspianskiego 27, 50-370 Wroclaw, Poland
* Author for correspondence and reprint requests
Z. Naturforsch. 59 c, 791Ð796 (2004); received July 22/August 8, 2004
Carrot seed oil is the source of the carotane sesquiterpenes carotol, daucol and β-caryo-
phyllene. These sesquiterpenic allelochemicals were evaluated against Alternaria alternata
isolated from the surface of carrot seeds cultivar Perfekcja, a variety widely distributed in
horticultural practise in Poland. Alternaria alternata is one of the most popular phytotoxic
fungi infesting the carrot plant. The strongest antifungal activity was observed for the main
constituent of carrot seed oil, carotol, which inhibited the radial growth of fungi by 65% at
the following concentration.
Key words: Carrot Seeds Oil, Carotol, Daucol, Antifungal Activity
Introduction
Nowadays there is a clear tendency towards the
utilisation of natural products, especially allelo-
chemicals, as alternative compounds for pest and
plant disease control, safe for humans and envi-
ronment. Therefore, the search of new natural
products including plant extracts, which might sub-
stitute synthetic agrochemicals or contribute to the
development of new agents for pest control, seems
to be important. It is well recognised that among
other plant products essential oils, rich in terpe-
noids and non-terpenoid compounds, possess vari-
ous and interesting allelopathic properties. Their
insecticidal action against specific pests and fungi-
cidal action towards some important plant patho-
gens have been recently reviewed (Isman, 2000).
Over the past decade a large volume of data
documenting the defence abilities of various seeds
with a long period of dormancy were accumulated
(Halloin, 1983; Harman, 1983; Kremer et al., 1984;
Ceballos et al., 1998; Özer et al., 1999). Among
them antimicrobial activity of chemicals exuded
from seeds and acting on soil-rhizosphere inter-
face was reported (Helsper et al., 1994). Quite sur-
prisingly, there is little information about the role
and importance of allelochemicals, which are pri-
mary constituents of seeds. The question whether
these compounds might play any role during the
0939Ð5075/2004/1100Ð0791 $ 06.00 2004 Verlag der Zeitschrift für Naturforschung, Tübingen · http://www.znaturforsch.com ·
D
storage, period of dormancy or at the very begin-
ning of seedling development and emergency still
remains unanswered. The fungal infections might
be considered as the main factor influencing the
health of plants at these early stages of their
growth and development. Therefore, it is not sur-
prising that many of the sesquiterpenoid com-
pounds are known from their antifungal activity
against plant pathogenic fungi, including the most
popular Alternaria sp. (Alvarez-Castellano et al.,
2001; Skaltsa et al., 2000). For example the sesqui-
terpene fulvoferruginin shows strong activity
against Gram-positive bacteria and significant an-
tifungal activity towards Paecilomyces varioti. An-
other carotane sesquiterpene, rugosal A, isolated
from Rosa rugosa, which is accumulated in the leaf
trichomes, shows antifungal activity against Cla-
dosporium herbarum (Ghisalberti, 1994).
Large varieties of compounds synthesised in
carrot tissues are known also for their allelopathic
activity. These include asarones (Guerin and Städ-
ler, 1984; Jasicka-Misiak and Lipok, 2000), chlo-
rogenic acid (Cole et al., 1988) and trans-2-nonenal
(Guerin and Ryan, 1980). In previous reports we
described phytotoxic activity of carrot seed oil and
its main terpenoic components (Jasicka-Misiak
et al., 2002).
To the best of our knowledge, the chemical com-
position and in vivo antimicrobial activities of
792 I. Jasicka-Misiak et al. · Antifungal Activity of Sesquiterpenes
carrot seed oil were investigated several times
(Guerin and Reveillere, 1985; Dwivedi et al., 1991;
Kilibarda et al., 1996;, Friedman et al., 2002). Al-
though, the information about allelopathic activity
of terpenes at all is massively accumulated in the
literature, the biological role of carrot seed sesqui-
terpenes is still poorly defined, especially in the
context of the low sensitivity of carrot seeds to
fungal infections.
The structures and chemical properties of two
most specific carrot seed sesquiterpenes, carotol
and daucol, were described (Sykora et al., 1961;
Hashidoko et al., 1992; Platzer et al., 1987; Bülow
and König, 2000), but nothing could be traced in
the literature about their antifungal activity. The
limited knowledge concerning the biological activ-
ity of daucol and carotol is surprising if con-
sidering the fact that they are present in carrot
seed oil in high amounts. Moreover, the oil is rela-
tively cheap and commercially available therefore
it can be treated as a valuable source of antifun-
gal substances.
In this work the novel, improved procedure of
isolation of carotol and daucol is reported and the
antifungal activity of these compounds against Al-
ternaria alternata, the most popular phytotoxic
fungus, was examined.
Materials and Methods
Starting material
Carrot seed oil was purchased from Augustus
Oils Ltd., London. β-Caryophyllene was pur-
chased from Aldrich, Poland. The commercially
available fungicide Funaben T containing thiuram
(45%) and carbendazim (20%) was produced by
Chemical Corporation Organika-Azot s.a., Poland.
Seeds of Daucus carota L. var. Perfekcja, collected
in 2001, were purchased from Torseed Co.,
Torun, Poland.
Isolation of carotol and daucol
Carotol. A portion (10 g) of the carrot seed oil
was subjected to silica gel (Merck Kieselgel 60;
0.063Ð0.2 mm particle size; 650 ¥25 mm ID) col-
umn chromatography. The column was eluted with
CH
2
Cl
2
, followed by 1% MeOH in CH
2
Cl
2
and
eluates were collected as 62 ¥20 ml fractions.
Each fraction was analysed by TLC (plate 5553,
Merck; solvent system benzene/EtOAc, 19:1 v/v,
developed by spraying with 0.5% anise aldehyde
in MeOH, heating at 105 C for visualisation), and
fractions 9Ð20 having one spot of R
f
0.86, charac-
teristic for carotol, were combined. The fractions
were evaporated to dryness (3.35 g) yielding a pale
yellow oil, which was identified by means of GC-
MS and
1
H NMR spectroscopy.
Daucol. It was obtained by a series of silica gel
column chromatographic steps. The first step was
the same as during carotol isolation. The fractions
were controlled by TLC, and fractions 46Ð55,
showing a similar spotting patterns zone (R
f
be-
tween 0 and 0.32), were combined. After evapora-
tion, the residue (0.19 g) was characterised by GC-
MS, showing the presence of 10 compounds. This
mixture was applied onto a silica gel column
(Merck Kieselgel 60; 0.063Ð0.2 mm particle size;
300 ¥15 mm ID). The column was eluted with
CH
2
Cl
2
/EtOAc (15:1 v/v). Eluates were collected
as 26 ¥20 ml subfractions, analysed by TLC, and
those containing daucol were combined (fractions
10Ð17). The obtained compound was recrystal-
lised from hexane at 4 C for 48 h and analysed by
GC-MS and
1
H NMR spectroscopy.
Structural studies
The analysis of the essential oil was performed
using a Hewlett Packard 6890 gas chromatograph
equipped with a FID detector. The diethyl ether
solution (1 µl) was injected into a HP-1 capillary
column (30 m ¥0.32 mm bonded-phase fused sil-
ica). The initial oven temperature was maintained
at 60 C for 2 min and then raised at 10 C min
Ð1
to 280 C. Helium was used as carrier gas. MS
analyses were performed on a quadrupole Hewlett
Packard 6897 instrument with ionisation at 70 eV.
The structure of the active compound was found
using a peak matching library search to published
standard mass spectra and by comparison with lit-
erature data.
The NMR experiments were carried out using
a Bruker DRX 300 MHz spectrometer. Chemical
shifts were referred to TMS (tetramethylsilane).
The proton and carbon assignments were per-
formed by means of COSY, TOCSY, HMQC,
DEPT-135 and HMQC-TOCSY experiments
using a spin lock time in the range 80Ð120 ms. The
TOCSY spectra were acquired with total spin-
locking time of 80 ms using the MLEV-17 mixing
sequence. In order to assign the carbon signal of
the daucol molecule the HMBC spectra were ad-
ditionally performed.
I. Jasicka-Misiak et al. · Antifungal Activity of Sesquiterpenes 793
Antifungal activity
Fungal bioassays in 9-cm Petri dishes were de-
signed to evaluate the influence of tested sub-
stances on mycelial growth of Alternaria alternata.
The mixtures of: carotol, caryophyllene and daucol
8:2:1 w/w/w (at the same weight ratio as it was
found in carrot seed oil and in carrot seeds); caro-
tol and daucol 8:1 w/w; carotol and caryophyllene
4:1 w/w; caryophyllene and daucol 2:1 w/w as well
as carrot seed oil were tested for antifungal activ-
ity. Appropriate amounts of above terpenoids
were mixed (before sterilization) with Czapek me-
dium to obtain the final concentration of 150 mg/l.
The commercially available fungicide Funaben T
was used as a control. The agar was allowed to
solidify and experiments were initiated by placing
6-mm fungal plugs taken from the growing mar-
gins of 9-day-old cultures, mycelial side down, on
Czapek medium. Plates were incubated at 24Ð
25 C. Radial growth of the strain was recorded
daily, by taking the mean diameter of colonies
from each plate. The same experiments were car-
ried out for individual compounds: carotol, daucol
and caryophyllene. All the experiments were per-
formed four times, with four replications with four
pure Czapek medium controls.
Data analysis
The data were subjected to analysis of variance
to test the significance of all factors examined in
each experiment. F test was done to determine the
homogeneity of error variances among runs. Treat-
ment means were separated using Tukey’s HSD
test at a 5% significance level (Statgraphics
Plus 5, 2000).
Results and Discussion
Carrot seed oil composition
The chemical composition of the carrot seed oil
was determined by GC-MS analysis. The results
are presented in Table I as percent of the total MS
ion current. The compounds are listed according
to their elution order. Monoterpenes and sesqui-
terpenes represent the major components of car-
rot seed oil. Out of 40 compounds detected in the
chromatogram of the carrot seed oil 33 were iden-
tified. In our studies, only those components which
were present in the oil in amounts higher than
0.1% have been taken into consideration.
Table I. Levels (peak area percent) of major compo-
nents of carrot seed oil purchased from Augustus Oils
Ltd.
Component Carrot seed oil
RT
a
RA
b
(%)
α-Thujene 3.06 1.90
α-Pinene 3.20 3.94
Camphene 3.43 0.92
β-Pinene 3.46 1.90
β-Myrcene 3.65 1.44
α-Terpinene 3.97 1.43
o-Cymene 4.03 1.34
Limonene 4.12 1.75
γ-Terpinene 4.49 1.43
Terpinolene 5.54 0.63
Linalool 6.08 0.51
Pinen-4-ol 7.32 0.42
Terpinen-4-ol 8.16 0.22
3-Carene 8.60 0.83
Neryl acetate 8.69 1.06
Calarene 8.80 3.23
Zingibren 8.99 2.13
α-Farnesene 9.10 3.35
β-Caryophyllene 9.18 10.66
α-Cedrene 9.35 2.74
α-Himachalene 9.44 0.55
β-Cubebene 9.57 0.53
α-Longipinene 9.73 0.76
Aromadendrene 9.93 1.92
β-Farnesene 10.08 4.03
Levomenol 10.18 0.34
Vitamin A aldehyde 10.33 0.66
Isolimonene 10.56 3.24
Caryophyllene oxide 10.97 4.34
Carotol 11.22 38.85
χ-Cadinene 11.49 0.25
Daucol 11.57 2.00
Total 99.30
a
Retention time on HP-1 column in minutes.
b
Relative area (peak area relative to total peak area).
The main components of the oil were carotol
(38.8%) and β-caryophyllene (10.7%), accompa-
nied by caryophyllene oxide (4.3%) and a second
daucane sesquiterpene alcohol namely daucol
which is present in significant amounts (2.0%).
The other identified volatile components have
been reported previously as constituents of an-
other organs of carrot (Seifert and Buttery, 1978;
Buttery et al., 1979; Kjeldsen et al., 2001). All of
them, except the sesquiterpenic alcohols carotol
and daucol are well-known compounds isolated
from many other plant sources. Especially, β-ca-
ryophyllene is a widespread sesquiterpene. Caro-
tol, daucol and β-caryophyllene comprised 51.5%
of the oil.
794 I. Jasicka-Misiak et al. · Antifungal Activity of Sesquiterpenes
OH
1
23
5
6
7
8
9
10
11
13
14
15
4
12
a
OH
O
1
23
5
6
8
9
10
11
12
14
15
b
4
Fig. 1. Structures of carotol (a) and daucol (b).
Isolation of carotol and daucol
The previously described methods for isolation
of carotol and daucol are based on fractional dis-
tillation (Sykora et al., 1961; Hashidoko et al.,
1992). In our opinion column chromatography is a
faster and simpler method since carotol was ob-
tained from carrot seed oil by only one step by
means of silica gel column chromatography with
high efficiency (99%). Fractions with daucol were
provided by the same column, however, they re-
quired further purification by means of additional
silica gel column chromatography. These three
chromatographic steps resulted in the isolation of
daucol of 99% purity.
1
H NMR and
13
C NMR spectra of carotol and
daucol (Fig. 1) assigned using the set of NMR ex-
periments (see Materials and Methods) are sum-
marized in Tables II and III.( The chemical shifts
of proton and carbon peaks for the carotol mole-
Table II.
1
H and
13
C chemical shifts of carotol in chloro-
form (CDCl
3
).
Number of
1
H
a13
C
a1
H
13
C
carbon
with
its proton
1C 49.09 49.08
2CH
2
1.70/2.26 38.62 1.70/2.26 38.62
3CH 5.32 122.13 5.32 122.13
4C 138.60 138.59
5CH
2
2.08 29.45 2.08 29.45
6CH
2
1.94 34.45 1.63/1.94 34.41
7C 84.55 84.54
8CH 1.80 52.54 1.79 52.53
9CH
2
24.39 1.52/1.68 24.39
10CH
2
1.3 39.45 1.29/1.57 39.45
11CH
3
0.95 21.47 0.95 21.47
12CH
3
1.67 25.23 1.67 25.24
13CH 1.80 27.58 1.81 27.59
14CH
3
1.00 24.04 1.00 24.05
15CH
3
0.95 21.38 0.94 21.38
7C-OH 1.14 1.14
a
Bülow and König, 2000.
Table III.
1
H and
13
C chemical shifts of daucol in chloro-
form (CDCl
3
).
Number of
1
H
a1
H
13
C
carbon with
its proton
1C 45.15
2CH
2
1.28/1.68 1.28/1.68 40.09
3CH 3.72 3.74 71.70
4C 85.22
5CH
2
1.36/1.86 1.37/1.86 29.50
6CH
2
1.55/2.15 1.58/2.15 41.15
7C 91.63
8CH 1.50 1.50 52.40
9CH
2
1.70 1.71 26.20
10CH
2
1.25/1.30 1.24/1.28 32.90
11CH
3
1.06 1.06 22.42
12CH
3
1.36 1.36 23.45
13CH 1.77 1.78 31.51
14CH
3
0.81 0.82 21.79
15CH
3
1.06 1.06 22.93
a
Platzer et al., 1987.
cule are in good agreement with those reported in
the literature (Bülow and König, 2000; Platzer
et al., 1987; Hashidoko et al., 1992). The proton
chemical shifts of daucol are also in good consis-
tence with only one up to now published data
(Bülow and König, 2000), however, additional as-
signments for carbon atoms were performed (Ta-
ble III).
Antifungal activity
The antifungal activity of carrot seed terpenoids
was tested on strains isolated from non-disinfected
and untreated seeds. Seven strains of fungi belong-
ing to the Alternaria family and one strain of
Acremonium were isolated from the surface of
carrot seeds. Thus, Alternaria predominated
among all the identified genera of fungi. Because
the phytopathogenic activity of Alternaria towards
carrot plants is well documented they were chosen
for further experiments.
At the start, we tested the effect of crude carrot
seed oil and appropriate mixtures of carotol,
caryophyllene and daucol in different weight ra-
tios (see Materials and Methods) on the growth of
Alternaria alternata. The composition of the mix-
ture of the three terpenoids was set at the same
ratio as it was found in crude oil. For comparison
the additional tests with the commercially avail-
able fungicide Funaben T were also performed.
The obtained results are shown in Fig. 2a. Carrot
I. Jasicka-Misiak et al. · Antifungal Activity of Sesquiterpenes 795
Fig. 2. The effect of tested mixtures (a)
and poor substances (b) at 150 mg/l on
mycelial growth of Alternaria alternata
0
1
2
3
4
5
6
7
8
9
10
11
12345678910
control
seed oil
daucol
carotol
Funaben T
Cultivation time [d]
Colony diameter [cm]
0
1
2
3
4
5
6
7
8
9
10
11
12345678910
control
carotol;caryophyllene (2:1 w/w)
carotol;caryophyllene (4:1 w/w)
carotol;daucol (8:1 w/w)
carotol;caryophyllene;daucol (8:2:1 w/w/w)
Funaben T
a
b
on solid media.
seed oil exhibited moderate inhibitory effects on
mycelium radial growth of Alternaria alternata
(20% of inhibition). Mixtures containing only ca-
rotol exhibited strong inhibitory activity. This ac-
tivity increased with an increasing content of caro-
tol in individual mixtures. The highest inhibition
of the growth of pathogenic fungi was determined
for a carotol and daucol mixture (66%) in which
the content of carotol was the highest (90%).
The experiments with individual substances,
namely carotol, caryophyllene and daucol were
carried out to find out, whether the observed ac-
tivity derives from the action of carotol only or
from a synergetic nature. The kinetics of inhibition
of Alternaria alternata by these terpenoids used at
the concentration 150 mgl/l in Czapek medium is
shown in Fig. 2b. It is clearly seen that these com-
pounds started to influence the fungal growth after
the third day of incubation (see the standard devi-
ation bars) and the differences in their action had
significantly grown with time. Carotol significantly
inhibited the growth of the fungi and reduced the
colony radial size by 65% at the 9
th
day of the
experiment. Quite different effects were observed
for daucol (second specific sesquiterpene alcohol
of the oil) where slight stimulation of the develop-
ment of Alternaria alternata was observed. Wide-
spread in various plants the sesquiterpene β-ca-
ryophyllene failed to have any effect. Therefore, it
can be concluded that carotol is the main agent
attributed for antifungal activity of carrot seeds.
The activity of carotol is nearly as strong as of the
commercially abailable fungicide Funaben T
(85%).
796 I. Jasicka-Misiak et al. · Antifungal Activity of Sesquiterpenes
Acknowledgements
This research was supported by Polish State
Committee for Scientific Research (Komitet Ba-
dan
´Naukowych) grants PBZ KBN Ð060/T09/
2001/37. Piotr Młynarz wishes to thank The Foun-
dations for Polish Science for scholarship.
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... Olejek eteryczny pozyskiwany z nasion składa się w głównej mierze z monoterpenów i seskwiterpenów, w tym karotolu, daukolu, β-kariofilenu, tlenku kariofilenu, których sumaryczna zawartość przekracza 50%. Ponadto, w olejku eterycznym znajdują się takie składniki jak: daucen, germakren D, β-bergamoten, β-bisabolen, sabinen, α-pinen [4,7,24]. Olejek eteryczny z kwiatów (baldachy z dojrzałymi nasionami) w przeważającej ilości zawiera zarówno monoterpeny węglowodorowe (hydrocarbon monoterpenes) (ok. ...
... Właściwości przeciwgrzybicze i herbicydowe olejku eterycznego wynikają z obecności w jego składzie karotolu, β-kariofilenu i tlenku kariofilenu. Zdaniem Jasickiej-Misiak najsilniejszą aktywność przeciwgrzybiczną wykazuje mieszanina karotolu i daukolu, gdzie karotol stanowi 90% składu [24]. ...
... Karotol, daukol wyizolowany z olejku eterycznego z nasion marchwi [24] Aktywność w stosunku do Cryptococus neoformans (MIC = 0,16 mL/mL), dermatofitów (MIC = 0,32-0,64 ml/ml) flukonazol (MIC: 16 mg/mL, 16-128 mg/mL) Olejek eteryczny [8] Aktywność w stosunku do drożdży (MIC = 0,6-5,0 mL/mL) Olejek eteryczny [12] Działanie przeciwutleniajace Całkowita zawartość polifenoli w ekstrakcie metanolowym z liści marchwi 82,07 mg/mL w przeliczeniu na kwas galusowy Ekstrakt z liści [15] Zdolność inhibicji rzędu 86% dla ekstraktu etanolowego o stężeniu 70%, całkowita zawartość fenoli 92% (mg GA/g suchego ekstraktu), zawartość flawonoidów 15,6% (mg luteoliny/g suchego ekstraktu) ...
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Daucus carota L. – marchew zwyczajna to pomarańczowe warzywo rozpoznawalne na całym świecie, należące do rodziny selerowatych (Apiaceae), używane w medycynie już od czasów antycznych. W Polsce można wyróżnić Daucus carota L. subsp. carota występującą w stanie dzikim, oraz gatunki uprawne Daucus carota L. subsp. sativus (Hoffm.) Arcang. var. sativus Hoffm (marchew uprawna o pomarańczowym kolorze), oraz Daucus carota L. subsp. sativus (Hoffm.) Arcang. var. atrorubens Alef tzw. marchew czarna (marchew uprawna odmiany czarnej). Korzeń, liście, kwiaty i nasiona z marchwi są powszechnie wykorzystywane do celów spożywczych, farmaceutycznych i kosmetycznych, ponieważ są źródłem makro-i mikro składników. Zawierają one wiele cennych fitoskładników takich jak: karotenoidy, pochodne kwasu hydroksycynamonowego, alkaloidy, flawonoidy fenole, oraz związki terpenowe. Pozyskuje się z nich olej, ekstrakty, maceraty, olejki eteryczne, a także związki chemiczne z grupy karotenoidów. Celem niniejszej pracy przeglądowej było przedstawienie i podsumowanie wiedzy dotyczącej składu chemicznego poszczególnych części marchwi oraz aktywności biologicznej głównych surowców pozyskiwanych z Daucus carota L. Zwrócono także uwagę na możliwe ich aplikacje w kosmetologii oraz bezpieczeństwo stosowania. W tym celu przedstawiono charakterystykę botaniczno-anatomiczno-ekologiczną gatunku Daucus carota. Omówiono skład chemiczny korzenia, liści i nasion z marchwi. Dokonano charakterystyki olejku eterycznego zarówno pod względem jego składu chemicznego jak i aktywności biologicznej. Przedstawiono metody otrzymywania oleju z nasion, jego właściwości fizykochemiczne oraz skład kwasów tłuszczowych. Całość zamyka wykaz surowców kosmetycznych z marchwi dostępnych na rynku. Wg bazy CosIng obejmuje on ekstrakty, hydrolaty, olejki eteryczne, soki, kultury komórkowe, protoplasty i hydrolizaty. Surowce te scharakteryzowano podając ich nazwy INCI, działanie, przeznaczenie oraz typ produktów w jakich są używane. Różnorodność pozyskanych surowców z marchwi zwyczajnej i ich wysoka aktywność biologiczna wynikająca z obecności taki związków jak: karotol, α-pinen, β-kariofilen, sabinen, tlenek kariofilenu octan geranylu, przekładają się na szeroki wachlarz zastosowań D. carota, czyniąc ją roślinną popularną i cenioną dla wielu gałęzi przemysłu w tym przemysłu farmaceutycznego, spożywczego i kosmetycznego.
... Silva Dias et al. (2014) also reported the antimicrobial activity of wild carrots. Jasicka-Misiak et al. (2004) reported that carrot contains sesquiterpene compounds which inhibited the sixty-five percent radial growths of fungi. ...
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Carrot (Daucus carota L.) is a nutrient-rich vegetable that is widely cultivated and consumed in Pakistan in both raw and processed form. Data on the proximate composition and natural occurrence of aflatoxins (AFs) in carrots and marketed carrot products is lacking in Pakistan and the risk exposure of AF has not been characterised before. Thus, the current study was designed to know the frequently consumed carrot products with per capita consumption, and risk assessment of AF through these products in various regions of South Punjab Pakistan. A survey was conducted with 125 respondents and appeared that raw carrot, fresh carrot juice, gajrella and pickle are the most frequently consumed marketed carrot products with per capita consumption i.e. 62.5, 46.6, 16.2 and 14.5 gday-1, respectively. Proximate analysis revealed that carrot root and processed carrot products contained 9.65-98.2% moisture, 0.23-0.60% ash, 6.2-14.1% carbohydrates, 0.31-0.80% protein, 0.40-3.7% fat and 1.4-4.20% fibre. AF analysis revealed that 36.67% of samples were contaminated with TAF. Thirty-five (35%) percent of samples were tainted with aflatoxin B1, and 13.33% of samples were contaminated with aflatoxin B2. All the samples of carrot root, fresh carrot juice and gajrella contained TAF levels less than the maximum limit (ML) (2 ppb) assigned by the European Union (EU). However, the entire AFB1 positive samples of carrot pickle contained AFB1 levels of more than 2 ppb exceeding the ML. Furthermore, daily dietary exposure of TAFs ranged from 0.11 to 1.27 ng/kg of body weight per day which relatively exceeds the permissible limit of 1 ng/kg of body weight per day as defined by the Joint FAO/WHO Expert Committee on Food Additives. This is the first prevalence and risk assessment report of AF in marketed processed carrot products in Pakistan. These baseline data are an initial step in the effort to deal with this significant food safety issue and the establishment of legislation for AF in marketed products is needed in Pakistan.
... A strategy to overcome the issues presented by GO can be the encapsulation of this compound in NLCs using environmentally friendly liquid vegetable oils and natural solid lipids to create a safe and effective carrier for its topical application [30][31][32]. Among the many natural-derived oils that can be chosen from, carrot seed oil has proved to be a good candidate for skin protection and disease prevention, as it has been proven to have not only antibacterial and antifungal activity but also antiaging potential due to its antioxidant activity (1-diphenyl-2-picrylhydrazyl and nitric oxide free radical scavenging) and SPF and has proven skin compatibility as well (no irritation) [33][34][35][36]. Another promising and natural-sourced oil in shea butter has proven to have several relevant activities as well, such as antibacterial, antifungal, wound and burn healing, and anti-inflammatory, antioxidant, and even chemopreventive action [37][38][39]. ...
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Carrot is an economically important vegetable crop worldwide. Its storage root, the consumed organ, varies broadly within the carrot germplasm, exhibiting different colors due to the accumulation of anthocyanin and carotenoid pigments, as well as extensive variation for phytochemicals composition and consumer-quality traits. Anthocyanins and other phenolics, carotenoids, polyacetylenes, and terpenes represent the major carrot nutraceutical classes. In recent years, the use of next-generation sequencing technologies has facilitated the application of “multi-omics” approaches, in combination with transgenics and classical genetic tools, for studying the genetics underlying the accumulation of these phytochemicals in the carrot root. In purple carrot, such approaches allowed the identification and mapping of simply inherited and quantitative trait loci (QTLs) conditioning anthocyanin pigmentation in different tissues and genetic backgrounds, and the discovery of key genes conditioning anthocyanin biosynthesis, glycosylation, and acylation. Glycosylation and acylation influence the chemical stability and bioavailability of anthocyanins, and therefore their potential use as food colorants or nutraceutical agents, respectively. Similarly, important advances were made for two major loci conditioning carotenoids accumulation in white, yellow, and orange roots, namely Y and Y2. With the sequencing of the carrot genome, a candidate for the Y gene involved in photosystem development and carotenoid storage was described, whereas fine mapping of Y2 drastically reduced the genomic region of interest to 650-kb, but a clear candidate was not identified. Another gene, Or, which regulates chromoplasts development, was associated with carotenoids presence in the carrot root. Besides these nonstructural genes, progress towards understanding the role of several carotenoid biosynthetic genes has been made. The genetics of carrot polyacetylenes is also becoming increasingly understood. Candidate fatty acid desaturase 2 (FAD2) genes with specific desaturase and/or acetylenase activities have been identified by QTLs analysis and proposed as catalyzers of different steps in the polyacetylene pathway, and their genomic organization was described. Similarly, gene members of the large carrot terpene synthase family were catalogued, partially associated with QTLs for characteristic carrot root monoterpenes like sabinene, and functionally characterized in vitro. This chapter reviews and discusses recent advances in genetics and genomics of the main carrot nutraceuticals.
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Chapter
Carrot is an economically important vegetable crop worldwide. Its storage root, the consumed organ, varies broadly within the carrot germplasm, exhibiting different colors due to the accumulation of anthocyanin and carotenoid pigments, as well as extensive variation for phytochemicals composition and consumer-quality traits. Anthocyanins and other phenolics, carotenoids, polyacetylenes, and terpenes represent the major carrot nutraceutical classes. In recent years, the use of next-generation sequencing technologies has facilitated the application of “multi-omics” approaches, in combination with transgenics and classical genetic tools, for studying the genetics underlying the accumulation of these phytochemicals in the carrot root. In purple carrot, such approaches allowed the identification and mapping of simply inherited and quantitative trait loci (QTLs) conditioning anthocyanin pigmentation in different tissues and genetic backgrounds, and the discovery of key genes conditioning anthocyanin biosynthesis, glycosylation, and acylation. Glycosylation and acylation influence the chemical stability and bioavailability of anthocyanins, and therefore their potential use as food colorants or nutraceutical agents, respectively. Similarly, important advances were made for two major loci conditioning carotenoids accumulation in white, yellow, and orange roots, namely Y and Y2. With the sequencing of the carrot genome, a candidate for the Y gene involved in photosystem development and carotenoid storage was described, whereas fine mapping of Y2 drastically reduced the genomic region of interest to 650-kb, but a clear candidate was not identified. Another gene, Or, which regulates chromoplasts development, was associated with carotenoids presence in the carrot root. Besides these nonstructural genes, progress towards understanding the role of several carotenoid biosynthetic genes has been made. The genetics of carrot polyacetylenes is also becoming increasingly understood. Candidate fatty acid desaturase 2 (FAD2) genes with specific desaturase and/or acetylenase activities have been identified by QTLs analysis and proposed as catalyzers of different steps in the polyacetylene pathway, and their genomic organization was described. Similarly, gene members of the large carrot terpene synthase family were catalogued, partially associated with QTLs for characteristic carrot root monoterpenes like sabinene, and functionally characterized in vitro. This chapter reviews and discusses recent advances in genetics and genomics of the main carrot nutraceuticals.
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From glandular trichome o f Rosa rugosa, three major sesquiterpene hydrocarbons [acora-3(4),7(15)-diene, carota-4(5),ll(12)-diene (isodaucene) and carota-l,4-diene], and two minor [daucene and acora-3(4),7(8)-diene] were identified. The presence o f these carotadienes, which are possibly precursors o f the corresponding C-14-oxygenated carotanoids, suggested that the formation o f the carotane skeleton follow ed by the regio-specific oxygenation at C-14 occur in the biosynthesis o f R. rugosa carotanoids.
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The effects of first generation carrot fly larval damage on chlorogenic acid concentration in carrots was investigated in a field experiment at Wellesbourne in 1985. In a separate experiment carrots grown in the absence of a resident population of carrot fly were also analysed for chlorogenic acid; these carrots maintained low concentrations of chlorogenic acid through the summer and autumn until low ground temperatures occurred from November to January. The relationship between chlorogenic acid concentration and damage by the first generation of carrot fly was described by a similar model to the one derived previously for late-generation damage but without the cultivar dependence. This may have been because first generation damage takes place in mid-summer when soil temperature is not sufficiently low for differential chlorogenic acid production by carrot cultivars. The model supports the hypothesis that carrot fly damage increases chlorogenic acid production which subsequently encourages further attack. The increase in acid production due to the low winter temperature may be the mechanism which, in turn, induced a differential cultivar response in carrots harvested during the winter.
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