52

Seasonal variation in the total phenol contents in cultivated and wild carqueja
[Baccharis trimera (Less) DC.]

1 2 2 1 1 2
SILVA, F.G. ; PINTO, J.E.B.P. ; NASCIMENTO, V.E. ; SALES, J.F. ; SOUCHIE, E.L. ; BERTOLUCCI, S.K.V.
1 2
Centro Federal de Educação Tecnológica de Rio Verde. Rio Verde, GO. 75901-970; Universidade Federal de
Lavras. Lavras, MG. 37200-000 e-mail: suzan@ufla.br

ABSTRACT: This work aimed at the assessment of the seasonal variation influence on the content
of total phenols in wild and cultivated populations of carqueja. The plants have been evaluated
every month for one year at 6:00 a.m, 12:00 p.m., and 6:00 p.m. The experimental delineation
was the randomized blocks, factorial 2x3x12 (2 populations, 3 harvest time, and 12 months), with
three replicates, totalizing 216 experimental units. The vegetal material was harvested, dehydrated,
and grounded at the extraction time. The analysis was performed using the spectrometric method.
The harvest time and the populations did not influence on the total phenol contents. Higher contents
of total phenols were found between May and October for the cultivated populations, and from
June to September for wild species. Lower contents were detected in the rainy season – higher
temperatures and months of intense vegetative growth.

Key words: Asteraceae, phenols, medicinal plant. Braccharis.

RESUMO: Variação sazonal no teor de fenóis totais em carqueja [Braccharis trimera

(Less) DC.] cultivada e silvestre. Este trabalho teve como objetivo avaliar a influência da variação
sazonal no teor de fenóis totais em populações silvestre e cultivada de carqueja. Realizaram-se
coletas mensais durante 12 meses e três horários (06:00, 12:00 e 18:00 h). O delineamento
experimental foi em blocos casualizados, fatorial 2x3x12 (2 populações, 3 horários de colheita e
12 meses), com 3 repetições, perfazendo 216 unidades experimentais. O material vegetal coletado
foi a parte aérea da planta. Estas foram colhidas, desidratadas e, no momento da extração,
trituradas. O método de análise utilizado foi o espectrofotométrico. O horário de colheita e as
populações não influenciaram no teor de fenóis totais. Maiores teores de fenóis totais foram
encontrados entre os meses de maio e outubro para a população cultivada e de junho a setembro
para a silvestre. Menores teores de fenóis totais ocorreram na época mais úmida, com maiores
temperaturas e no período de intenso crescimento vegetativo.

Palavras-chave: Asteraceae, fenóis, planta medicinal, Braccharis.

INTRODUCTION
The successive evolutionary adaptation from
plants to the soil was reached basically by the massive
formation of phenolic compounds. These substances,
besides assuming a structural role on the cell wall,
present large formation of non structural constituents
with several functions, as plant defense (allelopathic
and antifungal agents, for example). It is determined,
for instance, the wood durability and the flower color
establishment, which contribute significantly to certain
flavor (taste and smell). These functions and others
influenced by vegetable phenols are essential for the
continuous survival of all species of vascular plants
(Carvalho et al., 2004; Croteau et al., 2000; Robbers
et al.,1997; Scogings et al., 2004).
In relation to the role of phenols on plant
pathology interaction, researches have demonstrated
that that class of secondary metabolites has great
importance in this sense. The induction of synthesis
of phenolic compounds is a common characteristic
of the host-pathology interaction, and the phenol
specificity has been related to the host resistance. In
wheat leaves, resistant to Puccinia reecondita, ferulic

Recebido para publicação em 12/12/2005

Aceito para publicação em 15/02/2007

Rev. Bras. Pl. Med., Botucatu, v.9, n.3, p.52-57, 2007.


and p-coumaric acids were found (Southerton &
Deverall, 1990). Eucalyptus calphylla R.Br.
(Myrtaceae) treated with aminooxiacetate (PAL
inhibitor), altered the resistance response to
Phytophtora cinnamomi, modifying the lignin and
phenol concentration which presented phenotypic
characteristics of susceptibility. These results
evidence the relation between phenol and lignin
synthesis and the plant resistance to certain
pathologies. Studies have demonstrated the
involvement of peroxidases in the biosynthetic via of
poliphenols and lignin (Almeida et al., 1986).
The concentration of phenols in vegetable
tissues changes according to its biosynthesis rate,
storage and degradation. It can be influenced by the
hormonal balance or controlled directly by enzymes,
and also by the enzymatic substrate balance (Siqueira
et al., 1991). Other factors, as light, temperature,
carbohydrate content, nutrition and water status, can
influence on the phenol synthesis, despite its
concentration change in relation to the type of phenolic
composition, the plant species, the growth phase and
the part of the plant evaluated. Stress conditions can
also influence the releasing of poliphenols from the
vacuoles, as well as the synthesis of new phenols
(McConchie et al., 1994). For instance, groups of acid
phenols, as ferulic and p-coumaric acids, are
commonly found in roots, while isoflavonoids
compounds are predominantly observed in leaves and
less in petioles, stems and roots (Siqueira et al.,
1991).
Generally, the concentration of defense
chemical compounds, rich in carbon, as the phenols,
is correlated negatively with the growth rate and
reduction of the plant vigour. Also, the soluble phenols
as tannin may influence the diet option and the food
consumption due to an adstringent taste on the food
or reducing the nutrient availability after ingestion
(Scogings et al., 2004).
Souza (1998), working on Plantago major L.
(Plantaginaceaea), detected higher values of phenol
content in shoot than roots, where the difference was
more expressive during the reproductive phase. In the
root system, the phenol content did not alter under
different light levels and plant growth phases. During
the inflorescence maturity the phenol content
increased, but under 100% it was more precocious
than in cultivated plants at 50%. In relation to the
increase on the phenol content in leaves as in
inflorescence, it occurs due to the synthesis of
phenolic compounds, which reduce the oxidation of
auxins, or are directly involved in the flourishing
process.
Bucher et al. (2003) studied the phenol
content during the four seasons of the year in some
species at Córdoba region (Argentina). Among these,
Lycium cestroides Schltdl. (Solanaceae) presented
Rev. Bras. Pl. Med., Botucatu, v.9, n.3, p.52-57, 2007.
53
highest content among the seasons; Prosopis spp.
(Fabaceae) and Celtis tala Gillies ex Planch.
(Ulmaceae) did not show any differences among the
seasons while Schinus longifolius Speg.
(Anacardiaceae) presented the larger content in Spring
and Summer. The chemical composition of the plant
material also influenced the diet in Phytotoma rutila
which was combined, besides other factors, to the
phenol content.
Studies carried out in South Africa with
several plant species demonstrated, in a general way,
against the initial expectations, that the condensed
tannins and total phenols were higher in all species
during the growth season. That observation was
interpreted as an indication that a great strategy of
defense that potentially reduces the risk of active
apical buds lost to browsers. In the growth period,
Scutia myrtina Kurz (Rhamnaceae) and Acacia karroo
Hayne (Mimosoideae) presented an increase on the
total phenol content. Similar result was found with
Diospyros lycioides Desf. (Ebenaceae), Ehretia rigida
Druce (Boraginaceae); Grewia accidentalis (Tiliaceae)
and Rhus longispina Eckl & Zeyh. (Anacardiaceae)
(Scogings et al., 2004).
In Baccharis myriocephala Baker
(Asteraceae), the tannin content evaluated through
the quantification of total phenols increased at the
beginning of the flowering, while at the end of the
flourishing period, that content lowered (Castro et al.,
1999).
In B. dracunculifolia DC. (Asteracease) was
detected a larger tannin content during the reproductive
phase, compared to the nonreproductive phase. The
relation between the concentration of tannin and the
abundance of galls was not verified. The authors
suggested that the abundance of galls in that specie
is probably determined by another factor that changed
among the phenological phases, as other secondary
compounds or defense mechanisms were not
evaluated in the study (Espírito-Santo & Fernandes,
1998).
Capsicum annuum L. (Solanaceae) plants
were cultivated under three different irrigation levels
aiming to determine its effect on the phenylpropanoid
metabolism. The soluble phenolics and lignin content
in padrón pepper irrigate with intermediate levels of
water were higher than those grown under drought-
stressed. The results of that study suggest that the
environmental conditions, as drought stress present
a strong effect on the capsaicinoids accumulation in
padrón fruits. It is the result of competition between
the capsaicinoids biosynthesis and other
phenylpropanoid metabolites (Estrada et al., 1999).
The aim of this study was to quantify the total
phenol content of Baccharis trimera (Less) DC., in
wild and cultivated populations, in different months
and harvest time, in order to determine the best time
54
to the phenol production. Moreover, the influence of
the culture management on the content of that
compound was also evaluated.
MATERIAL AND METHOD
General conditions: the study was carried out
in two areas from March, 2003 to February, 2004, in
Lavras, MG (21º13’40”and 44º57’50” W GRW). The
cultivated plants were obtained at “Universidade
Federal de Lavras/Setor de Plantas Medicinais”, grown
in absence of weeds. The wild plants were at 20 km
far from the cultivated species, at “Três Barras” farm.
The latter place was chosen due to the natural
occurrence and abundance of Baccharis trimera.
Those plants were grown mixed with other wild
species. The plants were identified by Prof. Dr. Manoel
Losada Gavilanes of Departamento de Biologia,
Universidade Federal de Lavras (UFLA), Minas Gerais
State, Brazil. Voucher specimen is deposited in the
Herbarium at UFLA (code number 169933).
During one year, the shoot and inflorescences
(if present) were harvested every month at 06:00 a.m,
12:00 p.m and 6:00 p.m.
Phytochemistry analysis: the method of
analysis used was the spectrophotometric
(Association of Agricultural Chemist - AOAC, 1970).
Total phenol extraction: 100 mg of each dry sample
was submitted to three consecutive extractions with
methanol (3 mL) at 40 ºC, for 10 minutes. The extracts
were collected and filtered and the volume was
completed to 10 mL with methanol. For the standard
curve, tanic acid (P.A) and the Folin-Denis reagent
plus NaCO3 saturated solution was used to determine
the phenol concentration. After 30 minutes shaking,
the absorption was determined at 760 nm.The readings
TABLE 1. Chemical characterization of soil samples in cultivated and wild carqueja populations.
Rev. Bras. Pl. Med., Botucatu, v.9, n.3, p.52-57, 2007.
were made in a spectrophotometer (Hitachi U-2000),
using triplicates per sample. The results were
expressed in concentration of tanic acid 100 mL-1 of
sample and converted in % of tannin 100 mg-1 of
dehydrated plant sample, according to Castro et al.
(1999). That value was used on the variance analysis.
Environmental characterization: Soil
samples were collected in both areas in April, 2003,
and analyzed at “Departamento de Solos/UFLA”
(EMBRAPA, 1999) in relation to the chemical (Table
1) and physical properties (Table 2). Despite of the
similar soil pH in both areas, considerable differences
were observed between P and K content, where higher
values were detected on the cultivated area and a
higher Al concentration on the wild area.
The regional climate is the Cwa type,
according to the Köppen climatic classification, with
an annual precipitation mean of 1530 mm, 80%
concentrated from October to March. The dry season
extends from April to September. The maximum,
medium and minimum temperature and precipitation
during the period studied is found on Figure 1a-b. The
soil humidity was monitored by the gravimetrical
method, with triplicates samples per treatment in 12
collects. Each soil sample was gathered from 0 to 20
cm, around 4:00 and 5:00 p.m, on the plant harvest
days. The humidity of soil during the experiment can
be seen on Figure 1c.
Statistical analysis: the experimental design
was in randomized blocks, factorial 2x3x12 (2
populations, 3 harvest time and 12 months), with
triplicates, totalizing 216 experimental units. The data
were subjected to ANOVA and means compared by
Scott-Knott test (p< 0.05), using the statistical
program SISVAR 4.0 (Ferreira, 2000).

TABLE 2. Physical characterization of the soil samples
in cultivated and wild carqueja populations at UFLA,
Lavras, MG, 2005.
RESULT AND DISCUSSION
The total phenols content in Baccharis trimera
plants showed little variation among the populations,
months and harvest time. The content changed from
3.26%, in cultivated plants, harvested in November at
6:00 a.m and 7.32% in wild plants harvested in July
at 12:00 p.m (Table 3). The experiment was carried
TABLE 3. Total phenols content (%) in shoot of wild and cultivated carqueja, harvested in different months and
times.
Mean followed by same letters between carqueja populations, in each harvest time, do not differ (Scott-Knott, p<0.05).
* superior mean among months (Scott-Knott, p<0.05). Each value represents a mean of triplicates.
The total phenol content did not change
among the wild and cultivated populations,
independent of the harvest time. In relation to the
months, the largest content occurred between May
and October for the cultivated population with a mean
of 6.34%, and from June to September for the wild
population with a mean content of 6.76% (Table 3).
Larger content of total phenols occurred in months
with lowest temperatures, precipitation and soil
humidity (Figure 1 a-c). The lowest total phenols
content were found at the vegetative growth period.
Different results were found for B. myriocephala where
the beginning of the flowering coincides with the tannin
content increase phase (Castro et al., 1999).
According to Herms & Mattson (1992), there
is a tendency of chemical compounds rich in carbon,
as the phenols, to be negatively correlated with the
growth rate and the reduction of the plant vigour. It
Rev. Bras. Pl. Med., Botucatu, v.9, n.3, p.52-57, 2007.
55
out under greenhouse conditions evaluating the same
species, in “Campos dos Goytacazes-RJ”. The mean
content of total phenols was 2.59% (Freitas et al.,
2004) being lower values than those found in this
study. Castro et al. (1999) working with Baccharis
myriocephala verified that the tannin content
expressed through the total phenols content changed
considerably (from 0.26% to 12.18%). Generally, the
plants present a large flexibility in the production of
chemical compounds caused by the genetic variability
and physiological factors, in order to increase the
energy flow obtained on the first metabolism
(photosynthesis) for the secondary metabolism (used
in the drug-fabrication). These factors can explain the
differences among the total phenols content (Brown
Junior, 1988).
was also observed in carqueja plants. On the other
hand, not all of the plant species follow that pattern.
For example, S. myrtina and Acacia karroo showed
the highest content of phenols in the growing season.
Similar observation was verified for D. lycioides, E. rigid,
G. accidentalis and R. longispina (Anacardiaceae)
(Scogings et al., 2004).
In relation to the harvest time, there was no
statistical difference on the total phenol content in
any months during the year (data not showed).
When the interaction among the factors
evaluated (populations, harvest time and months) is
analyzed, the differences among months when
analyzed in relation to the harvest time were not verified
in any of them (Table 3). Those differences occurred
in cultivated plants only at 6:00 a.m. On the wild plants,
a difference was verified at 6:00 a.m and 6:00 p.m.
56
FIGURE 1. Maximum, intermediate and minimum
temperature (A), precipitation (B) and soil humidity
(C) of 0 to 20 cm in depth in areas with cultivated and
wild carqueja. Dates from March 2003 to February
2004.
CONCLUSION
a) The harvest time did not influence the total
phenols content;
b) Cultivated and wild plants were similar in
relation to the total phenols content.
ACKNOWLEDGMENT
Thanks to “Conselho Nacional de
Desenvolvimento Científico e Tecnológico (CNPq)” for
the doctorate scholarship given to the first author.
Rev. Bras. Pl. Med., Botucatu, v.9, n.3, p.52-57, 2007.
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