Journal of materials and JMES, 2017 Volume 8, Issue 1, Page 318-323

Environmental Sciences
ISSN: 2028-2508

Copyright © 2017, University of

Mohammed Premier - Oujda Morocco http://www.jmaterenvironsci.com/

Effect of Seasonal Variation on the Growth and Chemical Composition of

Cynara Cardunculus L. Plants

Hend E. Wahba1, Atef Z. Sarhan2, Adel B. Salama1,

Mahmoud A. Sharaf-Eldin3,1,*, Heba M. Gad1
1
Department of Medicinal and Aromatic Plants Research, National Research Centre, 12622-Dokki, Egypt
2
Ornamental Horticulture Dept., Faculty of Agriculture, Cairo University, Giza, Egypt
3
Sara Alghonaim Research Chair, Biology Department, College of Science and Humanities, Prince Sattam bin Abdulaziz
University (PSAU), 11942-Alkharj, Saudi Arabia

Received 01July 2016, Abstract

Revised 16Aug 2016, Cynara cardunculus L. (Family Asteraceae), is a highly nutritious plant contains
Accepted 20Sept 2016 medically active compounds such as flavonoid, polyphenol, caffeoylquinic acids.
During the few decades seasonal changes affect the production of crop worldwide. In
Keywords this study, we studied the changes in the vegetative growth and chemical
composition during the growing season from the first of February to the first of June.
 Photocatalysis, The samples were harvested monthly from the three replicates during 2009/2010 and
 Cynara, 2010/2011 seasons. The results obtained in this study showed changes in
 harvest, physiological factors (plant height, number of leaves) and leaves biomass (fresh and
 Polyphenols, dry weight) increased progressively with age of plant from February to June. But
 Flavonoids,
 Chlorogenicacid these increments were reached beyond in May at a significant level. The active
ingredient, carbohydrate, flavonoid and polyphenol increased gradually with age up
to May which gave the maximum value. While, caffeic acid, vanillic acid and
sharafeldin99@yahoo.com chlorogenic acid reached the maximum value during April month.

1. Introduction
Cynara cardunculus L. (Family Asteraceae) commonly known as cardoon, is an herbaceous perennial plant
originated from Mediterranean countries. This plant has many therapeutic potential as antidiabetic,
antimicrobial, cholagogue, choleretic and diuretic [1,2]. Two active compounds, chlorogenic acid and cynarin
(phenolic in nature) are found in the cardoon plant belongs to derivatives of caffeic acid. The leaves were also
used as a stimulant in liver cell regeneration, choleretic lipid-lowering and hepatoprotective [3-7]. The
importance of C. cardunculus is mainly referred to its content of the flavonoids and polyphenol. The metabolic
processes leading to accumulation of these active constituents in the plant are basically controlled by the
physiological age of the plant and the surrounding environmental conditions, as well as, the genetic factors.
Therefore, it is of great importance from the production point of view, to follow up the growth parameters and
chemical composition of the plant throughout the growing season. Hammouda et al. [8], found that the
flavonoid and polyphenol contents in C. scolymus (Romanian strain) cultivated in Egypt varied with plant age,
and plants 9- to 10-months-old were most suitable for pharmaceutical preparations. During seasonal changes,
many investigators reported that the content of phenolics and flavonoids varied with the developmental phase of
the plants. Djurdjevic et al. [9] observed that the maximum content of total phenolics was found in Conyza
Canadensis L. plants during the flowering and fruiting time in August rather than phenolics peaked during
elongation and excessive plant growth in May and June or in September. Phenolic acid (P-coumaric, ferulic, p-
hydroxybenzoic, vanillic and syringic) revelled from the stem have a maximum twice reached maximum in May
and in August. In a study on the effects of seasonal variation of the major secondary metabolites present in the
extract of Eremanthus mattogrossensis less (Asteraceae) leaves. The main compounds were identified and
quantified, and the metabolites were grouped by chemical class (caffeoylquinic acids, flavonoids, and
sesquiterpene lactone). Statistical analysis indicated a straight correlation between the quantity of metabolites
and seasonality, suggesting that environmental properties elicit important metabolic responses [10]. Also,

JMES, 2017, 8 (1), pp. 318-323 318

Thomsen et al. [11] observed that, the major alkamides in the roots of Echinacea purpurea were recorded its
lowest concentration in the mid of autumn and early winter while the total concentration of lipophilic
compounds in Echinacea pallida showed the same pattern. It is also noted that, all of the major active phenolic
acids in E. purpurea were maximum concentrations in spring. Keeping in view, the objective of this study was
designed and investigates the variation in the yield and chemical composition of C. cardunculus during the
vegetative cycle of the plant which determine the optimum time for its harvesting yield and quality ingredients.

2. Materials and Methods

The seeds of Cynara cardunculus were obtained from Dr. Helmut Junge, ABiTEP GmbH, Berlin, Germany,
providing the plant material, via Jelitto GmbH, Germany. This work was carried out during two successive
seasons 2009/2010 and 2010/2011 at the Experimental Farm of Agriculture Faculty, Cairo University, Giza.
This experiment was done to follow up the changes in the vegetative growth and chemical composition during
the growing season from the first of February to the first of June where samples were taken monthly from the
three replicates during 2009/2010 and 2010/2011 seasons.

2.1. Nature of soil

Samples from the soil were taken before cultivation, and were subjected for physical and chemical analysis in
Soil Science Department, National Research Centre according to method of Jackson [12]. Physical properties:
clay 22.0% (dry matter), silt 51.0%, sand 26.4%, organic matter 0.6%, soil texture (sandy loam). Chemical
properties: pH 8.0, E.C. (dS/m-1) 1.15, available N 1.40% (dry matter), available P 0.83%, available K 0.27%.
Cations: (Milliequivalent/L) Ca2+ 12.2, Mg2+ 3.7, Na+ 0.27, K+ 0.27. Anions: (Milliequivalent/L) CO32– 0.0,
HCO3– 1.1, Cl- 1.4, SO42- 13.5.

2.2. Soil preparation for cultivation

During both seasons, the soil was mechanically ploughed and planked twice. During preparation of the soil for
cultivation mixture of calcium superphosphate (15.5% P2O5) at the rate of 475 kg ha-1 as a source of phosphorus
was added and mixed well manually with the soil.

2.3. Cultivation procedures and maintenance

Seeds were sown in plastic bags of 23X18 cm in a medium of clay (1 sand: 1clay) under the sun screen. The
uniform healthy cardoon seedlings (60 day old) were transplanted into the field on the first week of November
in both seasons, at a distance 100 cm apart between plants into plots 6 m2 (3 X 2). Each plot contained 2 rows,
with 6 plants. All other horticultural practices were done as needed for the whole period of the growing season.
The plants were fertilized with 590 Kg ha-1 ammonium sulphate (20.5% N) as a source of nitrogen. The nitrogen
was applied as a side dressing at three equal additions. The first addition was after 8 weeks of transplanting and
the second was 6 weeks later, while the third was added after 6 weeks from the second addition. Potassium
sulphate (48% K2O) as a source of potassium at the rate of 355 Kg ha-1was added at two equal amounts, the first
dose was added with the second dose of nitrogen and a second was added with the third dose of nitrogen.

2.4. Data recorded

On the basis of phenology of cardoon we have analysed the following parameters in term of plant height,
number of leaves and biomass of leaves (fresh and dry weight) per plant.

2.5. Chemical analysis

2.5.1. Estimation of total carbohydrate
Total carbohydrates in the dried leaves were determined according to Dubois et al. (1956) [13]. Total
carbohydrates were calculated by using standard curve of glucose.

2.5.2. Estimation of total flavonoids

Determination of total flavonoids content in the dry leaves of C. cardunculus was determined by
spectrophotometer according to Kosalec et al. [14].

2.5.3. Estimation of total phenols

Phenols content was determined in the dry aerial parts by spectrophotometer according to Falleh et al. [15].

JMES, 2017, 8 (1), pp. 318-323 319

2.5.4. Estimation of chlorogenic acid, caffeic acid, and vanillic acid content in leaves of cardoon through High
Performance Liquid Chromatography (HPLC)
Using HPLC (Agilent Technologies, Palo Alto, CA, USA) chlorogenic acid, caffeic acid, and vanillic acid
content was determined in the aerial parts (dry matter) according to the method described by Sharaf-Eldin et al.
[16] and according to the modification reported by Wahba et al. [17].

2.6. Statistical analysis

This study was performed in a completely randomized manner with different treatment with three replicates.
Data of each season were statistically analysed by ANOVA [18]. The LSD (least significant difference) level at
5% was used to compare the means value according to Snedecor and Cochran [19].

3. Results and discussion

3.1. Growth parameters
3.1.1. Plant height cm plant-1
Table (1) compiles the data of growth parameters taken through the first and second seasons (2009/2010 and
2010/2011) from the first of February to the first of June. The results show that the plant height increased
gradually from February and attain the highest height (cm plant -1) in start of June, but the differences between
May and June were insignificant. On the other hand the differences between other months were significant. The
least value in this concern was obtained in February (88.33 and 93.33 cm plant -1) against the highest values
(108.33 and 113.33 cm plant-1) in June for the first and second seasons, respectively.

3.1.2. Number of leaves plant-1

Table (1) showed the same trend occurred in the case of plant height. The number of leaves increased
progressively with age of the plant from February (8.33 and 9.67) till June (21.0 and 20.33) for the first and
second seasons, respectively. Generally, it is noticed that the values of leaves number plant-1 in May and June
were insignificant.

Table 1: Effect of age and seasonal variation in plant height (cm) and number of leaves plant-1 of Cynara
cardunculus during 2009/2010 and 2010/2011 seasons.
Growth characters
Months
Plant height (cm) Number of leaves plant-1
1st season 2nd season 1st season 2nd season
February 88.33 93.33 9.33 9.67
March 91.67 95.00 10.33 11.33
April 96.67 101.67 17.00 16.67
May 106.67 111.67 20.33 21.33
June 108.33 113.33 21.00 20.33
LSD(0.05) 7.79 8.47 1.63 1.56

3.1.3. Fresh and dry weight of leaves g plant-1 and ton ha-1
From the data illustrated in Figure (1), the results reveal that the production of herb increased progressively and
gradually from February till June. The differences for fresh weight between February and other months were
significant, but the increased in the fresh weight through April and May months were insignificant as compared
to fresh weight of herb through June month. This trend was true during both seasons. In the same Figure (1), the
dry weight of herb shows a similar trend such as fresh weight. The fresh and dry weight of herb (leaves) ton ha-1
had similar of results of fresh and dry weight g plant -1. The treatments which encouraged the fresh weights were
the same which produced the high values of fresh and dry weight of leaves g plant-1.

3.2. Chemical composition

3.2.1. Total carbohydrate (%) in leaves
The results in Figure (2) show that the percentage of total carbohydrate increased gradually from February to
May, and attain the highest value in May where reached to 13.68% and 13.95% against 7.70% and 7.96% in
February for the first and second seasons, respectively. On the other words, the total carbohydrate content

JMES, 2017, 8 (1), pp. 318-323 320

increased with age from February till May, while the percentage of total carbohydrate in June decreased as
compared to May. The differences between the values of total carbohydrate % in February, March, April and
May were significant, but the difference between May and June was insignificant.

Fresh weight g/plant

Fresh weight g/plant
dry weight g/plant
3000 dry weight g/plant
Fresh and dry weight of leaves

2500 3000
2500
2000
2000
1500
1500
1000 1000
500 500
0 0
.Feb .Mar April .May .Jun .Feb .Mar April .May .Jun
Months Months
1st season 2nd season
Fig. 1. Effect of seasonal variation in fresh weight leaves g plant -1 and dry weight g plant-1 of Cynara
cardunculus during 2009/2010 and 2010/2011 seasons.

3.2.2. Total flavonoid in leaves mg g-1

The total flavonoid in leaves of C. cardunculus plant ranged between 2.93 to 7.81 mg g-1 in the 1st season and
3.56 to 7.96 mg g-1 in the 2nd season during the growth season (Figure 2). The results clear that, the total
flavonoids decreased progressively from February to June. The maximum value in this concern (7.81 and 7.96
mg g-1) was resulted from the leaves which harvested at the start of growth in February against the lowest value
in June (2.93 and 3.56 mg g-1) for the first and second seasons, respectively.

3.2.3. Total polyphenol in leaves mg g-1

Data in Figure (2) indicate that leaves of C. cardunculus were collected monthly from February till June. The
range of values of polyphenol content was (4.86 to 7.86 mg g-1) in the 1st season and (5.26 to 7.91 mg g-1) in the
2nd one. The maximum value of polyphenol in herb was in May (7.86 and 7.91 mg g-1), follow by (6.24 and 6.48
mg g-1) in April, then (6.17 and 6.42 mg g-1) in June. On other hand, the lowest value was obtained from the
sample of February. The differences between these values were significant in both seasons, except the values
between the samples of April and June during the second season.

Carbohydrate % Carbohydrate %
Flavonoid (mg/g) Flavonoid (mg/g)
Polyphenol (mg/g) Polyphenol (mg/g)
16 16
14
Chemical constituents

14
12 12
10 10
8 8
6 6
4 4
2 2
0 0
Feb. Mar. April May. Jun. Feb. Mar. April May. Jun.
Months Months
1st season 2nd season
Fig. 2.Effect of seasonal variation in percentage of the chemical content of Cynara cardunculus leaves during
2009/2010 and 2010/2011 seasons.

JMES, 2017, 8 (1), pp. 318-323 321

3.2.4. Total caffeic acid, vanillic acid and chlorogenic acid in dry leaves
The data of the second season only presented in Figure (3) is evident that production of leaves for both caffeic
acid, vanillic acid and chlorogenic acid were suitable in April, where gave the highest values for these
compounds; on the contrary the month of Feb. gave the lowest amount of these compounds. On the other words,
the total caffeic acid, vanillic acid and chlorogenic acid increased gradually from Feb. to April which reaching
28.19, 125.44, 535.68 mg 100g-1, respectively. On the contrary, the values of these compounds decreased during
May and Jun months. Our results are in agreement with the finding of Djurdjevic et al. [9] on Conyza
Canadensis L. plant, Gouvea et al. [10] on Eremanthus mattogrossensis and Thomsen et al. [11] on Echinacea
purpurea. Conyza Canadensis L. is also used in Moroccan Rif for its phyto-therapeutic actions against liver
inflammation [20].

Caffic acid (mg/100g) Vanilic acid (mg/100g)

chlorogenic acid (mg/100g)
600
Chemical constituents

500
400
300
200
100
0
Feb. Mar. April May. Jun.
Months
2nd season

Fig. 3.Effect of seasonal variation of total caffeic acid, vanillic acid and chlorogenic acid content of Cynara
cardunculus leaves during 2010/2011 season.

Conclusions
According to our results, we recommend to the cardoon's growers aiming to obtain the highest values of fresh
and dry weight of leaves along with the high values of chemical constituents, to harvest the cardoon leaves
during May under Giza's, Egypt climatic conditions, and/or in general before flowering stage. While, aiming to
obtain high values of carbohydrate and polyphenol, should harvest cardoon leaves during May, but for flavonoid
content, harvesting should be during February. If aiming to obtain high values of caffeic acid, vanillic acid and
chlorogenic acid, should harvest cardoon leaves during April.

Acknowledgments-The authors are thankful to Dr. Helmut Junge for providing the seeds of cardoon plant, which he kindly
purchased from Jelitto GmbH, Germany.

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