Abstracts
This research investigates effects of salicylic acid (an abiotic elicitor) on the antioxidant activity and betacyan production from leaves of Alternanthera tenella cultured in vitro was evaluated. Plants were grown in a liquid MS medium and vermiculite substrate. After 35 days salicylic acid was added to the medium. Content of betacyanins, total phenols and flavonoids and non-enzymatic antioxidant capacity were determined in leaves of A. tenella after 0, 12, 36 and 48h of treatment. After 36h, concentration of betacyanins and total phenols increased. On the other hand, the increase of the treatment time caused a slight decrease in total flavonoids and reduced the DPPH free radical activity. As result the antioxidant activity of the leaves of A. tenella is promoted by salicylic acid and can be attributed to the increase in betacyanin content, which are compounds with recognized antioxidant action.
Este trabalho investiga o efeito do ácido salicílico (um elicitor abiótico) sobre a atividade antioxidante e produção de betacianinas em folhas de Alternanthera tenella cultivada in vitro. As plantas foram cultivadas em meio MS líquido e vermiculita como substrato. Após 35 dias, o ácido salicílico foi adicionado ao meio. Conteúdo de betacianinas, fenóis totais e flavonoides e a capacidade antioxidante não-enzimática foi determinada em folhas de A. tenella após 0, 12, 36 e 48h de tratamento. Após 36h, a concentração de betacianinas e fenóis totais aumentaram. Em contrapartida, o aumento no tempo de exposição causou uma ligeira diminuição nos teores de flavonoides totais e inibição da atividade do radical livre DPPH. Como resultado, a atividade antioxidante de folhas de A. tenella é promovida pelo ácido salicílico e pode ser atribuída ao aumento do conteúdo de betacianina, os quais são compostos com ação antioxidante reconhecida.
Alternanthera tenella; DPPH; flavonoides totais; compostos fenólicos; betacianina.
INTRODUCTION:
The production of free radicals in living organisms is controlled by various antioxidant compounds of endogenous origin, from diet or other sources. The radicals formed from antioxidants are not subject to the chain propagation reaction, but are neutralized by reaction with another radical, forming stable products or being recycled by another antioxidant (ATOUI et al., 2005).
In this context, many studies have shown that phenolic compounds from plants exhibit
high antioxidant potential and, in addition to antioxidant enzymes, directly capture
reactive oxygen species (ANDRADE et al., 2007ANDRADE, C.A. et al. Determinação do conteúdo fenólico e avaliação da
atividade antioxidante de Acacia podalyriifolia A. Cunn ex. Don
Leguminosae-Mimosoidae. Revista Brasileira de Farmacognosia, v.17, p.231-235, 2007.
Available from: <http://www.scielo.br/scielo.php?pid=S0102-695X200700
0200017&script=sci_arttext>. Accessed: Nov. 10, 2011. doi:
10.1590/S0102-695X2007000200017.
http://www.scielo.br/scielo.php?pid=S010...
).
Natural antioxidants, such as phenolic compounds, inhibit lipid peroxidation and
lipooxygenase in vitro (SOARES,
2002SOARES, S.E. Ácidos fenólicos como antioxidantes. Revista de Nutrição,
v.15, p.71-81, 2002. Availablefrom:
<http://www.scielo.br/scielo.php?pid=S1415-52732002000100008&script=sci_abstract&tlng=pt>.
Accessed: Nov. 01, 2013.doi: 10.1590/S1415-52732002000100008.
http://www.scielo.br/scielo.php?pid=S141...
). The activity of the phenolic compounds is mainly due to their reducing
properties. These properties are important in neutralization and capture of free
radicals and chelation of transition metals, acting in the initiation and propagation
stages of oxidation. The intermediaries formed by the action of phenolic antioxidants
are relatively stable due to high electronic delocalization on the π-system of the
aromatic ring (CHUN et al., 2005CHUN, S.S. et al. Phenolic antioxidants from clonal oregano (Origanum
vulgare) with antimicrobial activity against Helicobacter pulori. Process
Biochemistry, v.40, p.809-816, 2005. Available from:
<http://www.sciencedirect.com/science/article/pii/S0032959204001189>. Accessed:
Nov. 15, 2011. doi: 10.1016/j.procbio.2004.02.018.
http://www.sciencedirect.com/science/art...
). Interest in
betacyanins has also grown since the characterization of their anti-radical activity.
These compounds have been widely used as a food additive and constituent in
pharmaceuticals and cosmetics due to their natural colorant properties and absence of
toxicity (STRACK et al., 2003STRACK, D. et al. Recent advances in betalain research., Phytochemistry
v.62, p.247-269, 2003. Available from:
<http://www.sciencedirect.com/science/article/pii/S0031942202005642>. Accessed:
Nov. 12, 2011.doi: 10.1016/S0031-9422(02)00564-2.).
Betacyanins are found in a number of plants, including Alternanthera tenella
Colla, an herbaceous plant commonly found in Brazil (popularly known as
"apaga-fogo") (FERREIRA et al.,
2003FERREIRA, E.A. et al. Estudos anatômicos de folhas de espécies de
plantas daninhas de grande ocorrência no Brasil. IV - Amaranthus spinosus,
Alternanthera tenella e Euphorbia heterophylla. Planta Daninha, v.21, p.263-271,
2003. Available from:
<http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-83582003000200012>.
Accessed: Oct. 02, 2011. doi: 10.1590/S0100-83582003000200012.
http://www.scielo.br/scielo.php?script=s...
). This species has antibacterial and antifungal activities and infusions
are orally taken for the treatment of inflammations and infections (BIELLA et al., 2008BIELLA, C.A. et al. Evaluation of immunomodulatory and anti-inflammatory
effects and phytochemical screening of Alternanthera tenella Colla (Amaranthaceae)
aqueous extracts. Memórias do Instituto Oswaldo Cruz, v.103, p.569-577, 2008.
Available from:
<http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762008000600010>.
Accessed: Dec. 04, 2011. doi: 10.1590/S0074-02762008000600010.
http://www.scielo.br/scielo.php?script=s...
). This species is considered a
weed (SIQUEIRA, 1995SIQUEIRA, J.C. Fitogeografia das Amaranthaceae Brasileiras. São
Leopoldo: IAP, 1995. 131p.), usually eliminated in
plantations, and thus becoming as endangered one (DECRETO 42.099, 2002DECRETO N.42.099, de 31 de dezembro de 2002. Declara as espécies da
flora nativa ameaçadas de extinção no estado do Rio Grande do Sul e dá outras
providências. Porto Alegre: Palácio Piratini, 2002.).
Species of the genus Alternanthera contain various biologically active
compounds, including betalains (betacyanins and betaxanthins), ecdysteroids, flavonoids,
saponins, and triterpenes (FERREIRA & DIAS,
2000FERREIRA, E.O.; DIAS, D.A. A methyllenedioxyflavonol from aerial parts
of Blutaparon portulacoides. Phytochemistry, v.53, p.145-147, 2000. Available from:
<http://www.sciencedirect.com/science/article/pii/S0031942299003908>. Accessed:
Oct. 10, 2011.doi: 10.1016/S0031-9422(99)00390-8.
http://www.sciencedirect.com/science/art...
).
The micropropagation of medicinal plants is a widely used technique for the production
of active compounds and natural products for the pharmaceutical industry. Compared with
traditional methods, in vitro techniques have many advantages, such as
independence of seasonal variation for massal propagation, identification and production
of clones with the desired characteristics, and the ability to manipulate the
microenvironment to increase production of the metabolite of interest (EL-TARRAS et al., 2012EL-TARRAS, A. et al. In vitro multiplication of the important medicinal
plant, harmal. Journal of Medicinal Plants Research, v.6, p.3586-3590, 2012.
Available from:
<http://www.academicjournals.org/journal/JMPR/article-abstract/F6A8D3F21276>.
Accessed: Nov. 04, 2013. doi:10.5897/JMPR12.315.
http://www.academicjournals.org/journal/...
). The use of elicitors in
in vitro culture has been one of the most effective strategies to
increase the formation of secondary metabolites, including the application of certain
hormones associated with stress in plants (JALLEL et al., 2009; PEROTTI et al., 2010PEROTTI, J.C. et al. Produção de betacianina em erva-de-jacaré cultivada
in vitro com diferentes concentrações de sulfato de cobre. Ciência Rural, v.40,
p.1874-1880, 2010. Available from:
<http://www.scielo.br/scielo.php?pid=S0103-84782010000900004&script=sci_arttext>.
Accessed: nov. 04, 2013.doi:10.1590/S0103-84782010000900004.
http://www.scielo.br/scielo.php?pid=S010...
).
Salicylic acid is a phenolic phytohormone and considered as efficient chemical elicitor.
This compound is involved in the signaling systems, inducing enzymes to catalyze the
formation of defense compounds such as polyphenols and alkaloids (VAN LOON, 1997VAN LOON, L.C. Induced resistance in plants and the role of
pathogenesis-related proteins. European Journal of Plant Pathology, v.103, p.753-765,
1997. Available from:
<http://igitur-archive.library.uu.nl/bio/2001-1220-112648/UUindex.html>.
Accessed: Sept. 27, 2011.
http://igitur-archive.library.uu.nl/bio/...
). When applied exogenously, elicitors can
systemically trigger the expression of a defense genes set that is naturally activated
by pathogen infection, stimulating the synthesis of various plant metabolites (ATSUSHI et al., 2007ATSUSHI, O. et al. Elicitor induced activation of the methyterythritol
phosphate pathway toward phytoalexins biosynthesis in rice. Plant Molecular Biology,
v.65, p.177-187, 2007. Available from:
<http://www.springerlink.com/content/5128776012668444/>. Accessed: Nov. 12,
2011. doi: 10.1007/s11103-007-9207-2.
http://www.springerlink.com/content/5128...
). This work describes the
influence of salicylic acid on the production of secondary metabolites and antioxidant
capacity of leaves of Alternanthera tenella cultured in
vitro.
MATERIALS AND METHODS:
A. tenella was collected in the City of Pelotas (Brazil). Its taxonomy was
confirmed by the Amaranthaceae identification key and was cataloged in the
PEL Herbarium under the number 25.26. Leaves of the plant were established in
vitro on MS medium (MURASHIGE &
SKOOG, 1962MURASHIGE, T.; SKOOG, F. A revised medium for rapid growth and bioassays
with tobacco tissue cultures. Physiologia Plantarum, v.15, p. 473-497, 1962.
Available from:
<http://onlinelibrary.wiley.com/doi/10.1111/j.1399-3054.1962.tb08052.x/>.Accessed:
Jun. 11, 2011.doi: 10.1111/j.1399-3054.1962.tb08052.x.
http://onlinelibrary.wiley.com/doi/10.11...
), without growth regulators, and were sub-cultured to obtain a
sufficient number of individuals for the experiment. Nodal segments of approximately 1cm
in length were removed and inoculated in flasks containing liquid MS and vermiculite -
used as a substrate to enable increased absorption of the elicitor. This procedure was
performedin a laminar flow chamber under aseptic conditions. The flasks were then placed
in the growth room at 25±2°C, under a 16h photoperiod and photon flux density of 48μmol
m-2 s-1. After 35 days, 15mL of elicitor (salicylic acid) was
added to the vermiculite at a concentration of 400µM. Leaves were collected at time
periods of 0, 12, 36, and 48h of the addition of the elicitor, and were stored in an
ultrafrezeer (-70°C) for later analysis of content of betacyanins, total phenolics, and
total flavonoids and antioxidant capacity. The choice of concentration was based on
experiments conducted in parallel (data not shown).
To determine betacyanin content, of fresh leaves (100mg) were macerated with celite in
distilled (5mL) water and centrifuged at 13632g and 4°C for 25min. The
assays were performed with the supernatant and absorbances were registered at 536 and
650nm in a spectrophotometer (Ultrospec 2100 Pro, Amersham Biosciences(r)),
as described by CAI et al. (1998)CAI, Y. et al. Characterization and quantification of betacyanin
pigments from diverse Amaranthus species. Journal of Agricultural and Food Chemistry,
v.46, p.2063-2070, 1998. Available from:
<http://pubs.acs.org/doi/abs/10.1021/jf9709966>. Accessed: Oct. 27, 2011. doi:
10.1021/jf9709966.
http://pubs.acs.org/doi/abs/10.1021/jf97...
. Betacyanin
concentration was calculated taking into account the coefficient of molar extraction for
amaranthine (5.66x104) and the results were expressed as mg of amaranthine
100g FM-1.
Total flavonoid concentration was determined by the colorimetric method described by
ZOU et al. (2004)ZOU, Y.P. et al. Antioxidant activity of a flavonoid-rich extract of
Hypericum perforatum L. in vitro., Journal of Agricultural and Food Chemistry v.52,
p.5032-5039, 2004. Available from:
<http://pubs.acs.org/doi/abs/10.1021/jf049571r?mi=xg2m3d&af=R&pageSize=20&publication=40026026&searchText=PMS-NADH>.
Accessed: Nov. 09, 2011.doi: 10.1021/jf049571r.
http://pubs.acs.org/doi/abs/10.1021/jf04...
. The absorbance reading was
performed in a spectrophotometer at 510nm, using aluminum chloride at 5% (m/v) in
methanol (WU & NG, 2008WU, S.J.; NG, L.T. Antioxidant and free radical scavenging activites of
wild bitter melon (Momordica charantia Linn. var. abbreviate Ser.) in Taiwan.
LWT-,Food Science and Technology v.41, p.323-330, 2008. Available from:
<http://www.sciencedirect.com/science/article/pii/S0023643807001181>. Accessed:
Sept. 27, 2011. doi: 10.1016/j.lwt.2007.03.003.
http://www.sciencedirect.com/science/art...
). The analysis
was performed using methanol extract (25µg mL-1) prepared from dry leaf
matter (250mg), which remained in of 70% methanol (10mL) for 24h.
Content of phenolic compounds was determined by Folin-Ciocalteu method, modified
according to JENNINGS (1981)JENNINGS, A.C. The determination of dihydroxy phenolic compounds in
extracts of plant tissues. Anal Biochemistry, v.118, p.396-398, 1981. Available from:
<http://www.sciencedirect.com/science/article/pii/000326978190600X>. Accessed:
Jun. 21, 2011.doi: 10.1016/0003-2697(81)90600-X.
http://www.sciencedirect.com/science/art...
. Fresh leaf material
(100g) was macerated, added to methanol:chloroform:water (12:5:3) solution (4mL). The
mixture was then placed into centrifuge tubes placed in the dark for 24h. Samples were
centrifuged for 10min at room temperature at 7000g. The supernatant was then collected
and the precipitate again centrifuged with methanol:chloroform:water (12:5:3) solution
(4mL) under the same conditions. The collected supernatant was joined to the first
centrifuging and, to each extract (4mL), chloroform (1mL) and water (1.5mL) were added,
followed by further centrifugation. Ultrapure water (500µL) and 1N Folin-Ciocalteu
reagent (500µL) were added to supernatant (500µL). After 15min, alkaline reagent (5mL),
consisting of sodium hydroxide (0.1N) and sodium carbonate (0.1N), were allowed to stand
for 60min. After this period, readings were performed in a spectrophotometer at 760nm,
using water as a blank. Phenic acid was used as standard for the construction of the
analytical curve at concentrations of 0-200µg mL-1, at intervals of 20µg. The
results were expressed in µg of phenic acid gFM-1.
Antioxidant activity was analyzed by the DPPH method (BRAND-WILLIAMS et al., 1995BRAND-WILIAMS, W. et al. Use of a free radical method to evaluate
antioxidant activity. Food Science and Technology, v.28, p.25-30, 1995. Available
from: <http://www.sciencedirect.com/science/article/pii/S0023643895800085>.
Accessed: Oct. 10, 2011. doi: 10.1016/S0023-6438(95)80008-5.
http://www.sciencedirect.com/science/art...
), which is based on the capture of the DPPH
radical (2.2-diphenyl-1-picryl-hydrazyl) by antioxidants, producing a decrease in
absorbance at 515nm. DPPH methanolic solution (60μM) was prepared to exhibit present
absorbance at 515nm between 0.6 and 0.7: 3.9mL DPPH solution and 0.1mL of the methanolic
extracts at 25µg mL-1 were added to test tubes. To assess the free radical
scavenging activity, the percent inhibition of DPPH in relation to the control sample
(methanol + DPPH 60μM) was calculated by the following equation: % inhibition of DPPH =
[(A0 - A1) / A0 x 100], where: A0 = absorbance of control and A1 = absorbance of the
sample (MOLYNEUX, 2004MOLYNEUX, P. The use of the stable free radical diphenylpicrylhydrazyl
(DPPH) for estimating antioxidant activity. Journal of Scienceand Technology, v.26,
p.211-219, 2004. Available from:
<http://rdo.psu.ac.th/sjst/journal/26-2/07-DPPH.pdf>. Accessed: Sept. 27, 2011.
http://rdo.psu.ac.th/sjst/journal/26-2/0...
).
The experimental design was completely randomized, consisting of four treatments (exposure time to salicylic acid), with three replicates. The experimental unit was represented by five flasks containing four explants per flask. The results were assessed by analysis of variance (ANOVA) and the means compared by Tukey's test at 5% error probability using the statistical software WinStat (MACHADO & CONCEIÇÃO, 2002).
RESULTS AND DISCUSSION:
Salicylic acid affects the secondary metabolism of plants (BOONSNONGCHEEP et al., 2010;
KORSANGRUANG et al., 2010) and has been widely studied in relation to signaling
mechanisms and responses to pests and diseases (FUJITA et al., 2006). In the present
study, exposing A. tenella to 400µM salicylic acid significantly influenced
the betacyanin content of the leaves. Time periods of 36 and 48h were sufficient to
stimulate the accumulation of these compounds (14.6 and 13.9mg of amaranthine
100gMF-1, respectively) when compared to the control (3.7mg of amaranthine
100gMF-1) (Figure 1A). However, the
observed concentrations of betacyanins were much lower than those found by KLEINOWSKI et al. (2014)KLEINOWSKI, A.M. et al. Pigment production and growth of Alternanthera
Plants cultured in vitro in the presence of tyrosine. Brazilian Archives Biology and
Technology, v.57, p. 253-260, 2014. Available from:
<http://www.scielo.br/pdf/babt/v57n2/cd_1037.pdf>. Accessed: Apr. 05, 2014.doi:
10.1590/S1516-89132013005000012.
http://www.scielo.br/pdf/babt/v57n2/cd_1...
in the stems of the same
species: 36.95mg of amaranthine 100g MF-1 with tyrosine elicitation (75µM).
This response might be due to the type of plant material and elicitor used in the
analyses. For example, betacyanins may be preferentially stored in the stem of A.
tenella.
Betacyanins content (A), total flavonoids content (B), total phenolic compounds content (C) and inhibition of the DPPH radical by methanolic extracts (D) from plants of Alternanthera tenella subjected to different exposure times to salicylic acid. Time periods labeled with the same letters do not differ significantly.
The success of an elicitor depends on the type and duration of treatment and
concentration and kind of elicited compound. For example, VAKIL & MENDHULKAR (2013)VAKIL, M.M.A.; MENDHULKAR, V.D. Salicylic acid and chitosan mediated
abiotic stress in cell suspension culture of Andrographi spaniculata (burm.f.) nees.
for and rographolide synthesis. International Journal of Pharmaceutical Sciences and
Research, v.4, p.3453-3459, 2013. Available from:
<http://www.ijpsr.com/V4I9/21%20Vol.%204,%20Issue%209,%20September%202013,%20IJPSR,%20RA%202604,%20Paper%2021.pdf>.
Accessed: Nov. 01, 2013 doi:10.13040/IJPSR.0975-8232.4(9).3453-59.
http://www.ijpsr.com/V4I9/21%20Vol.%204,...
investigated the effect of two
elicitors, salicylic acid and chitosan, on cultured cells of the medicinal plant
Andrographis paniculata. Astronger response was observed in the
production of secondary metabolites using chitosan in the treatment, although the exact
mechanism underlying this metabolic effect was unable to determine.
In the present study, salicylic acid elicited an increase in betacyanin synthesis in A. tenella. Nevertheless, increased exposure to the elicitor did not increase the concentration of flavonoids, possibly because the common precursor of both compounds, tyrosine, was preferentially converted into betacyanins.
Betacyanin is a type of betalains, hydrophilic pigments responsible for the coloration
of fruits, flowers, roots and leaves of plants belonging to the order Caryophyllales
(CASTELLANOS-SANTIAGO &YAHIA, 2008CASTELLANOS-SANTIAGO, E.; YAHIA, E.M. Identification and quantification
of betalains from the fruits of 10 Mexican prickly pear cultivars by high-performance
liquid chromatography and electrosprayionization mass spectrometry., Journal of
Agricultural and Food Chemistry v.56, p.5764-5778, 2008. Available
from:<http://pubs.acs.org/doi/abs/10.1021/jf800362t>. Accessed: Nov. 01,
2013.doi: 10.1021/jf800362t.
http://pubs.acs.org/doi/abs/10.1021/jf80...
).
Betalains and anthocyanins (flavonoids) are two different chemical classes of pigments
and are never found together in the same plant (GANDÍA-HERRERO et al., 2005GANDÍA-HERRERO, F. et al. Betaxanthins as pigments responsible for
visible fluorescence in flowers. Planta, v.222, p.586-593, 2005. Available from:
<http://link.springer.com/article/10.1007%2Fs00425-005-0004-3?LI=true#>.
Accessed: Oct. 13, 2011.doi: 10.1007/s00425-005-0004-3.
http://link.springer.com/article/10.1007...
). Such exclusiveness may have an explanation at
the biochemical level - the relevant enzymes for the production of anthocyanins are not
certainly expressed in betalain-producing plants (BROCKINGTON et al., 2011BROCKINGTON, S.F. et al. Complex pigment evolution in the
Caryophyllales. New Phytologist, v.190, p.854-864, 2011. Available from:
<http://www.ncbi.nlm.nih.gov/pubmed/21714182>. Accessed: Nov. 01,
2013.
http://www.ncbi.nlm.nih.gov/pubmed/21714...
).
The highest means of flavonoid contents were observed in the control plants and plants
after 12h exposure to the elicitor (6088.7 and 5932.9µg of quercetin g MF-1,
respectively), with a significant decrease (18%) for plants experiencing longer periods
of elicitation (36 and 48h) (Figure 1B). This is
in contrast to YU et al. (2006)YU, Z.Z. et al. Salicylic acid enhances jaceosidin and syringin
production in cell cultures of Saussurea medusa. Biotechnology Letters, v.28,
p.1027-1031, 2006. Available from:
<http://link.springer.com/article/10.1007%2Fs10529-006-9035-5?LI=true>.
Accessed: out. 04, 2011.doi: 10.1007%2Fs10529-006-9035-5.
http://link.springer.com/article/10.1007...
, who recorded a
gradual increase in total flavonoids over time, reaching 96% at 48h of treatment when
compared to control. However, the concentration of the elicitor was lower than that used
in the current work. HOUHUA et al. (2007)HOUHUA, L. et al. Maize Lc transcription factor enhances biosynthesis of
anthocyaninas, distinct pro anthocyanidins, and phenylpropanoids in apple (Malus
domestica Bords). Planta, v.226, p.1243-1254, 2007. Available from:
<http://link.springer.com/article/10.1007/s00425-007-0573-4>. Accessed: Oct.
25, 2011.doi: 10.1007/s00425-007-0573-4.
http://link.springer.com/article/10.1007...
observed significant increases in the synthesis of phenylpropanoids in Malus
domestica treated with salicylic acid. These results suggest that salicylic
acid induces the accumulation of compounds derived from phenylpropanoids due to the
increase in activity of the phenylalanine ammonia-lyase (PAL) enzyme. PAL is known to
play an important role in the transduction process, stimulating the synthesis of these
compounds and regulating the expression of defense genes (YU et al., 2006YU, Z.Z. et al. Salicylic acid enhances jaceosidin and syringin
production in cell cultures of Saussurea medusa. Biotechnology Letters, v.28,
p.1027-1031, 2006. Available from:
<http://link.springer.com/article/10.1007%2Fs10529-006-9035-5?LI=true>.
Accessed: out. 04, 2011.doi: 10.1007%2Fs10529-006-9035-5.
http://link.springer.com/article/10.1007...
).
Alternanthera tenella probably deviates from their secondary metabolite synthesis, where the shikimate acid pathway would lead to increased production of tyrosine. The accumulation of this amino acid would trigger the synthesis of betacyanin compounds with confirmed antioxidant activity.
A decrease in phenolic compound content was observed after 48h exposure to the elicitor
(1039.8µg g FM-1), having the maximum synthesis after 36h of elicitation
(2378.2µg of phenic acid g FM-1) with 400mM salicylic acid. However, after
12h of exposure there was no significant difference in phenolic compound content between
the treatment and the control (1657µg of phenic acid g FM-1) (Figure 1C). These results are somewhat different to
those of a recent study in Solanum melongena L. using different types of
elicitors and different exposure times (MANDAL,
2010MANDAL, S. Induction of phenolics, lignin and key defense enzymes in
eggplant (Solanum melongena L.) roots in response to elicitors. African Journal of
Biotechnology, v.9, p.8038-8047, 2010. Available from:
<http://www.academicjournals.org/ajb/PDF/pdf2010/22Nov/Mandal.pdf>. Accessed:
Nov. 17, 2011.doi: 10.5897/AJB10.984.
http://www.academicjournals.org/ajb/PDF/...
). The previous showed that salicylic acid was the most efficient elicitor
at triggering phenolic compound synthesis after 48h exposure - although a gradual
decrease in synthesis was observed over longer time periods (MANDAL, 2010MANDAL, S. Induction of phenolics, lignin and key defense enzymes in
eggplant (Solanum melongena L.) roots in response to elicitors. African Journal of
Biotechnology, v.9, p.8038-8047, 2010. Available from:
<http://www.academicjournals.org/ajb/PDF/pdf2010/22Nov/Mandal.pdf>. Accessed:
Nov. 17, 2011.doi: 10.5897/AJB10.984.
http://www.academicjournals.org/ajb/PDF/...
).
An assay of DPPH radical capture is a very useful tool for screening compounds for antioxidant potential. The analysis of antioxidant activity of A. tenella by the DPPH method indicated that exposure to the elicitor significantly increased the antioxidant potential of the extract. However, no statistical difference was observed between exposure times, indicating that plant extract has good antioxidant activity (Figure 1D).
The detoxifying effect of phenolic compounds on the DPPH radical has been well described
in literature (e.g. HEIM et al., 2002HEIM, K.E. et al. Flavonoid antioxidants: chemistry, metabolism and
structure-activity relationship. Journal of Nutritional Biochemistry, v.13,
p.572-584, 2002. Available from:
<http://www.aseanfood.info/articles/11014997.pdf>. Accessed: Oct. 28, 2011.
doi: 10.1016/S0955-2863(02)00208-5.
http://www.aseanfood.info/articles/11014...
). For
example, LEUNG & SHUI (2002)LEUNG, L.P.; SHUI, G. An investigation of antioxidant capacity of fruits
in Singapore markets. Food Chemistry, v.76, p.69-75, 2002. Available from:
<http://www.sciencedirect.com/science/article/pii/S0308814601002515>. Accessed:
Sept. 04, 2011.doi: 10.1016/S0308-8146(01)00251-5.
http://www.sciencedirect.com/science/art...
used the
DPPH method to characterize the antioxidant activity of 27 different fruits (with
extracts rich in phenolic compounds) found in the public markets of Singapore. A
detoxifying effect on the DPPH radical was also observed in extracts from 33 Chinese
medicinal plants containing phenolic compounds (TAN et
al., 2004TAN, S.Y. et al. Caracterization of antioxidant and antiglycation
properties and isolation of active ingredients from traditional Chinese medicines.
Free Radical Biology & Medicine, v.15, p.1575-1587, 2004. Available from:
<http://www.sciencedirect.com/science/article/pii/S0891584904002783>. Accessed:
Oct.02, 2011. doi: 10.1016/j.freeradbiomed.2004.03.017.
http://www.sciencedirect.com/science/art...
).
The increase in antioxidant activity observed in the present study is related to the
increased synthesis of betacyanins, since these compounds show high antioxidant activity
- as already reported in studies with sugar beet (STINTZING & CARLE, 2004STINTZING, F.C.; CARLE, R. Functional properties of anthocyanins and
betalains in plants, food, and in human nutrition. Trends in, Food Science and
Technology v.5, p.19-38. 2004. Available from:
<http://www.sciencedirect.com/science/article/pii/S0924224403001638>. Accessed:
Oct. 22, 2011. doi: 10.1016/j.tifs.2003.07.004.
http://www.sciencedirect.com/science/art...
). Indeed, betacyanins are among the ten most
powerful antioxidants due to their characteristic structural conformation. Betacyanins
have even been linked to the prevention of some types of cancer, including skin and
liver cancer, presumably as a consequence of their antioxidant properties (LILA, 2004LILA, M.A. Plant pigments and human health. In: DAVIES, K.S. (Ed.).
Plant pigments and their manipulation. 3.ed. Austrália:Blackwell Publishing Ltd,
2004. Cap. 14, p.248-274.).
CONCLUSION:
A. tenella has a significant pharmacological potential. Antioxidant activity of leaves of A. tenella can be significantly increased by the addition of an elicitor, such as salicylic acid, to the culture medium.
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Publication Dates
-
Publication in this collection
Oct 2014
History
-
Received
26 June 2013 -
Accepted
04 Mar 2014