Download O A RIGINAL

735
Journal of Applied Sciences Research, 9(1): 735-742, 2013
ISSN 1819-544X
This is a refereed journal and all articles are professionally screened and reviewed
ORIGINAL ARTICLES
Response of Calendula officinalis L. Plants to Foliar Application of Gibberellic Acid and
Mixture of Some Micronutrients
1
Mohamed, G.F. and 2Ebtsam, M.M. Abdella
1
Agriculture Botany Department and 2Horticulture Department, Fayoum Univ., Egypt
ABSTRACT
The present work was carried out in the Experimental Farm of Faculty of Agriculture, Fayoum University,
in two successive seasons 2010/2011 and 2011/2012. This work aimed to investigate the effect of gibberellic
acid and mixture of some micronutrients (Mn, Zn and Fe), as well as, their interaction on growth, flowering,
chemical constituents and anatomical structure of Calendula officinalis L. plants. The obtained results clearly
showed that increasing gibberellic acid concentration produced the highest significant or insignificant increasing
in plant height, branches number and fresh weight of herb, as well as, number of inflorescences/plant,
inflorescence diameter and inflorescences fresh weights/plant compared with control plants, in both seasons.
Relationship between GA3 applied and leaves or flowers macro and micronutrients contents was positive and so
total carbohydrates and pigments in leaves and flowers. Foliar application of micronutrients mixtures at the
highest concentration 400 ppm produced the highest significant records of the above-mentioned vegetative or
flowering traits. Pronounced enhancing influences on leaf and flower N, P, K, Mn, Zn, Fe, total carbohydrates
and pigments were obvious as a consequence of spraying micronutrients mixtures at any concentration
compared with control plants. The interaction of micronutrients mixtures and GA3 concentrations seemed to
reflect some positive significant effects on vegetative, flowering and chemical composition compared with
untreated plants. Generally, spraying micronutrients mixtures at 200 or 400 ppm combined with GA3 at 400
ppm was remarked in this respect. Anatomical properties of main stem were investigated and the highest
increase in stem section diameter was recorded at the interaction of 400 ppm GA3 and 200 ppm of mixture of
micronutrients.
Key words: Calendula officinalis, GA3, micronutrients
Introduction
Micronutrients play an important role in improving the vegetative growth and flowering of plants. Iron, zinc
and manganese are some of these elements which had a stimulative effect on growth of several plants. Such as,
Selim et al. (2001) on Calendula officinalis, Refaat and Balbaa (2001) on lemongrass plants, Rady et al. (2005)
on amaryllis plants, Ebtsam Abdella et al. (2006) on Polianthes tuberosa, Hassanain et al. (2006) on
chamomile plants, Naguib et al. (2007) on Ruta graveolens Yousef et al. (2010) on chamomile and Kumar and
Haripriya (2010) on nerium plants and concluded that spraying the plants with Zn, Mn or Fe increased all
vegetative growth characters and flowering, in addition increased chlorophyll content (a and b), total
carotenoids, N, P, K, Zn, Mn and Fe content.
Plant Growth regulators play an active role in improving plant vegetative growth. Gibberelic acid is one of
these substances which promote the vegetative growth of a wide range of ornamental or medicinal plants.
Several investigators studied these effects. For instance, Ebtsam Abdella and Rady (2005) on Asparagus
sprengeri, Rawia et al. (2006) on croton plants, Shah and Samiullah (2007) on black cumin, Azzaz et al. (2007)
on Calendula officinalis, Faten et al. (2008) on Hemerocallis auantiaca, Kazaz, and Karaguzel (2010) on
Solidags hybrida and Gehan and Mona (2011) on Palanites aegyptiaca plants, revealed that GA3 treatments
enhanced vegetative, flowering growth and chemical composition.
Materials And Methods
A field experiment was carried out during two successive seasons of 2010/2011 and 2011/2012 at the
Experimental Farm of Faculty of Agriculture, Fayoum University aiming to study the effect of gibberellic acid
and mixture of some micronutrients (Mn, Zn and Fe), as well as, their interaction on growth, flowering,
chemical constituents and plant anatomical of Calendula officinalis L. plants.
Corresponding Author: Ebtsam M.M.Abdella, Horticulture Department, Fayoum Univ., Egypt
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J. Appl. Sci. Res., 9(1): 735-742, 2013
Seeds of marigold (Calendula officinalis L.) were obtained from the Research Center of Medicinal and
Aromatic Plants, Giza, Egypt. Seeds were sown in nursery on 15th of September 2010 and 2011 seasons at the
Experimental area in Faculty of Agriculture, Fayoum University. The seedlings were transplanted after 45 days
from sowing date in the field on one side of the rows at a distance of 30 cm apart. The experimental plots were
(1.5*2.0 m) with three rows at a distance of 65 cm between rows. The plot contained 16 plants. Between every
two plots, there was a ridge 50 cm wide.
All experimental units received a constant level of NPK fertilizers (1:1:1) where ammonium sulphate (20.5
%N), calcium super phosphate (15% P2O5) potassium sulphate (48% K2O) were added at 4 g/plant divided in
two equal doses. The first and second editions were added after 3 and 6 weeks from transplanting, respectively.
The treatments comprised spraying the plants two times: 30 and 45 days after transplanting with gibberellic acid
at the rates 0.0, 100, 200 and 400 ppm, as well as, foliar application of mixture of micronutrients at the rates 0.0,
100, 200 and 400 ppm were sprayed one week after each edition of gibberellic acid. Few drops of Triton B were
added to the spray solution to serve as a wetting agent. The respective source of Mn, Zn and Fe was the
commercial fertilizer containing Mn, Zn or Fe at 13, 13, 13%,respectively, in the chelated form where the
chelating agent was citric acid. Control plants were sprayed with distilled water.
The experimental design was factorial experiment in complete randomized block design with five replicates
and three plants for each one.
Data recorded:
1. Vegetative growth characters:
In terms of plant height (cm), number of branches on the main shoot/ plant and fresh weight of herb/plant.
2. Floral traits:
Number of inflorescences/plant were counted, inflorescence diameter (cm) and inflorescences weights/plant
were determined.
3. Chemical composition:
At the end of the first season, leaf plastid pigments, i.e., concentration of total chlorophyll and total
carotenoids mg/100 g f.w. leaves by 80% acetone (at the ages 6 weeks from planting), were determined
according to the methods described by Cherry, 1973). Total carotenoids in flowers was carried out according to
Saric et al. (1967). In dry leaves and inflorescence total carbohydrates % was estimated colorimetrically as
outlined by Dubois et al, (1956), some macro and micronutrients were determined [total nitrogen was
colorimetrically determined by using orange G dye, according to method described by Hafez and Hichelson;
(1981), phosphorus was colorimetrically determined according to Jackson (1973), potassium was estimated
using a Flame-photometer Perkin–Elmer model 52 with acetylene burner as described by Page et al. (1982), Fe,
Zn and Mn concentrations were determined using a Zeiss Atomic Absorption AASS spectrophotometer
according to Page et al. (1982).
4. Anatomical study:
For anatomical study samples were taken at the end of first season (2010/2011) from stem. Samples were
killed and fixed in F.A.A. solution (10 ml formalin + 5 ml glacial acetic acid + 50 ml ethyl alcohol 95% + 35 ml
distilled water) for 72 hours, then dehydrated and cleared in n-butyl alcohol series, and embedded in paraffin
wax of 56-58ºC m.p. Cross sections of 20µ thick were cut, using a rotary microtom, adhesived on slides by
"Haupt's adhesive" then stained with the crystal violet–erythrosin combination, cleared in carbol xylene and
mounted in Canada balsam (Nassar and El-Shhar, 1998).
The obtained data were statistically analyzed according to the different treatments were achieved using
Least Significant Difference test (L.S.D.) at p= 0.05 (Snedecor and Cochran, 1980).
Results and Discussion
1. Vegetative growth characters:
Concerning the effect of micronutrients data in Table (1) reveal, in both seasons, the highest concentration
of micronutrients mixture (400 ppm) produced the highest values of all vegetative growth characters compared
to the other concentrations, as well as, untreated plants, with significant or insignificant differences between
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J. Appl. Sci. Res., 9(1): 735-742, 2013
mean values. Increase in vegetative characters due to the micronutrients can be attributed to improved root
system of plants resulting in absorption of more water and nutrients and its utilization. Moreover, micronutrients
activate several enzymes (catalase, peroxidase, alcohol dehydrogenase, carbonic dehydrogenase, tryptophane
syntheses, etc.) and involved themselves in chlorophyll synthesis and a lot of physiological activities by which
plant growth and development are encouraged (Prabhat and Arora, 2000). This results are in agreement with
those attributed by Selim et al. (2001) on Calendula officinalis, Refaat and Balbaa (2001) on lemongrass plants,
Rady et al. (2005) on amaryllis plants, Ebtsam Abdella et al. (2006) on Polianthes tuberosa, Hassanain et al.
(2006) on chamomile plants, Rawia et al. (2006) on croton plants and Naguib et al. (2007) on Ruta graveolens.
They concluded that using Fe, Zn or Mn individual or in mixtures affected positively on vegetative growth.
From data presented in Table (1) it is clear that in both seasons of studied, gradual increase of GA3
concentrations followed by gradual increase of vegetative growth characters (i.e. plant height, number of
branches/plant and fresh weight of herb/plant) with significant differences compared with the control plants. The
above-mentioned results may be due to the fact that GA3 may cause cell elongation by the induction of enzymes
that weaken the cell walls. Also, the mechanism by which gibberellins might stimulate cell elongation is that the
hydrolysis of starch resulting from the production of GA3 increased mobilization of starch in cotyledons by
increasing amylase activity (Kaur et al., 2000). The obtained data agree with that of Ebtsam Abdella and Rady
(2005) on Asparagus sprengeri, Rawia et al. (2006) on croton plants, Shah and Samiullah (2007) on black
cumin, Azzaz et al. (2007) on Calendula officinalis, Faten et al. (2008) on Hemerocallis auantiaca, Kazaz, and
Karaguzel (2010) on Solidags * Hybrida and Gehan and Mona (2011) on Palanites aegyptiaca plants. They
mentioned that GA3 treatments enhanced vegetative growth of plants.
Table 1: Response of Calendula officinalise L. plants to gibberellic acid and mixture of some micronutrients application on vegetative growth during the first and second seasons (2010\
2011 & 2011\2012).
M.N. pm
Plant height (cm)
No. of branches/ stem
Fresh weight/ plant (g)
First season (2010/2011)
0.0
100
200
400
Mean
0.0
100
200
400
Mean
0.0
100
200
400
Mean
GA3 ppm
0.0
74.67
90.66
92.00
80.00
84.33
10.00
5.67
10.66
11.67
9.5
155.00
557.00
447.33
506.33
416.42
100
80.33
96.33
90.33
99.67
91.67
6.00
19.00
11.00
18.67
13.67
422.00
332.33
561.67
547.00
465.75
200
77.00
77.67
92.33
100.33
86.83
7.67
15.33
17.33
15.67
14.0
378.33
421.67
459.00
436.67
423.92
400
88.67
92.33
106.33
99.33
96.67
13.67
14.00
20.00
14.67
15.59
508.33
417.33
450.00
567.00
485.67
Mean
80.17
89.25
95.25
96.67
9.34
11.83
14.75
15.17
365.92
432.08
479.50
514.25
L.S.D.5%
5.23
215.44
M.N.
18.27
2.62
107.72
GA3
9.13
6.04
248.77
21.09
M.N.*GA3
Second season (2011\2012)
0.0
52.33
86.33
85.67
94.67
79.75
9.67
6.67
14.67
14.00
11.25
117.67
578.33
423.33
453.33
393.17
100
79.67
72.67
85.00
78.67
79.00
8.33
15.67
11.00
16.00
12.75
451.67
496.67
386.67
413.33
437.09
200
88.00
76.67
88.00
81.33
83.50
13.00
14.00
18.67
13.00
14.67
420.00
383.33
596.67
568.33
492.08
400
85.00
95.67
94.00
113.33
97.00
14.67
14.67
14.00
15.67
14.75
495.00
466.67
615.00
703.33
570.00
Mean
76.25
82.84
88.17
92.00
11.42
12.75
14.56
14.67
371.09
481.25
505.42
534.58
L.S.D.5%
177.76
4.87
M.N.
17.27
88.88
5.62
GA3
8.63
205.26
5.62
19.94
M.N.*GA3
M.N.= mixture of micronutrient (Mn+ Zn+ Fe) , GA3 = Gibberellic acid
Regarding to the effect of the interaction between each of different concentrations of GA3 and
micronutrients mixtures. Spraying the foliage of Calendula officinalis plants with GA3 at 400 ppm combined
with micronutrients mixtures at 200 or 400 ppm produced the highest significant records of plant height
compared to the other interaction treatments, in the first or second seasons, respectively. The highest significant
records of branches number/\ plant were enhanced due to the interaction between GA3 at 400 ppm and
micronutrients mixture at 200 ppm or GA3 at 200 ppm combined with micronutrients mixture at 200 ppm, at the
two respective sesons. While, the highest significant weights of marigold plants were promoted resulted in the
interaction between micronutrients mixture at 400 ppm combined with GA3 at 400 ppm in both seasons
compared with the other interaction treatments.
2. Flowering traits:
Regarding the effect of micronutrient mixtures, significant increase in all studied of flowering traits were
revealed with raising the mixture of micronutrients rate up to 400 ppm, in both seasons of studied, Table (2).
These increments may be attributed to the positive effect of the micro-elements on most metabolic processes
such as carbohydrates, proteins, phosphate RNA and ribosome formation (Mostafa et al., 1997). Rady et al.
(2005) on amaryllis plants, Ebtsam Abdella et al. (2006) on Polianthes tuberosa, Hassanain et al. (2006) on
chamomile plants, claimed that foliar application with Fe, Zn or Mn individual or in mixtures enhanced
flowering growth.
Data arranged in Table (2) exhibited that gradual increase number of inflorescences/plant, inflorescence
diameter and inflorescences weights/plant records were parallel with gradual increase of GA3 concentration,
i.e., the highest concentration of GA3 (400 ppm) produced the highest significant records of the above-
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J. Appl. Sci. Res., 9(1): 735-742, 2013
mentioned flowering characters, in both seasons of study. The effect of GA3 on flower growth measurements it
may be due to the fact that gibberellins play a role in flowering, probably it is further elaborated into florigen by
the plant. Hence, gibberellins can not be the same substance as florigen but at least it may act as its precursor.
The propounder of (Florigen concept) florigen but made up of two substances, namely gibberellins and
anthesins. The latter are considered to be nitrogen rich compounds (Macleod and Millar, 1962). These results
are on line with Azzaz et al. (2007) on Calendula officinalis and Kazaz and Karaguzel (2010) on Solidags
hybrida. They concluded that pronounced enhancing influences on flowering growth was obvious as a
consequence of spraying GA3 at any concentration.
Table 2: Response of Calendula officinalise L. plants to gibberellic acid and mixture of some micronutrients application on flowering during the first and second seasons (2010\ 2011 &
2011\2012).
M.N. ppm
No. of inflorescences/ plant
Inflorescence diameter (cm)
Fresh weight of inflorescences/plant
GA3 ppm
0.0
100
200
400
Mean
L.S.D.5%
M.N.
GA3
M.N.*GA3
0.0
93.3
98.2
103.8
115.8
102.78
100
108.1
131.7
116.4
96.1
113.08
200
142.7
110.1
145.0
123.7
130.38
400
122.8
145.6
122.0
184.1
143.63
Mean
89.7
88.48
121.8
129.93
0.0
5.3
5.6
5.7
5.3
5.48
First season (2010\ 2011)
100
200
400
5.6
5.6
5.3
5.9
5.4
6.3
5.7
5.7
6.5
5.3
7.0
6.0
5.63
5.93
6.03
5.0
5.37
5.27
5.17
5.20
0.81
0.41
0.94
Second season (2011\2012)
5.77
5.5
4.83
5.76
5.17
6.06
5.33
5.5
6.27
5.27
6.83
5.83
5.53
5.75
5.75
9.41
4.70
10.86
0.0
92.0
105.67
136.0
119.67
113.34
100
96.67
130.0
100.33
141.67
117.17
200
102.67
113.0
137.0
121.0
118.42
400
114.0
94.0
117.67
180.0
126.42
Mean
101.34
110.67
122.75
140.59
L.S.D.5%
M.N.
9.44
GA3
4.72
M.N.*GA3
10.90
M.N.= mixture of micronutrient (Mn+ Zn+ Fe) , GA3 = Gibberellic acid
0.86
0.43
1.00
Mean
5.45
5.80
5.90
5.90
0.0
160.6
180.0
156.5
143.1
160.05
100
145.9
149.1
173.8
173.2
160.50
200
211.8
193.4
234.0
217.5
214.18
400
189.4
210.1
184.8
301.2
221.38
Mean
176.93
184.60
187.28
208.75
184.97
210.00
183.33
304.07
220.59
172.17
182.20
185.96
207.60
11.80
5.90
13.62
5.28
5.59
5.59
5.78
150.50
178.60
158.83
141.17
157.28
144.27
149.07
171.0
171.0
158.84
208.93
191.13
230.67
214.17
211.23
11.41
5.70
13.17
The interaction effects between different concentrations of GA3 and the mixture of micronutrients on all
studied of flowering traits were fluctuated significantly or insignificantly compared with control plants. The
highest records of number of inflorescences/plant or fresh weight of inflorescences/plant were resulted in due to
spraying the plants with micronutrients mixture at 400 ppm combined with GA3 at 400 ppm in both seasons.
While, the highest values of inflorescence diameter were recorded resulted in the interaction between GA3 at 400
ppm and micronutrients mixture at 200 ppm, in both seasons.
3. Chemical constituents:
As seen in Tables (3,4,5 and6), increasing micronutrients mixture application up to the highest rate (400
ppm) correspondingly significant increasing of P, Fe, Zn and total carbohydrates contents in leaves and flowers,
as well as, total chlorophyll, Mn and K contents in leaves or N content in flowers. However, micronutrients
mixture at 200 ppm was favorable and significant attained the highest mean value of N and total carotenoids in
leaves and K, Mn and total carotenoids in flowers. Generally, treated the plants with any concentration of
micronutrients mixture gave the highest contents of the above mentioned records compared with control plants.
These results may be attributed to the positive effect of the micro-elements on most metabolic processes such as
carbohydrates, proteins, phosphate RNA and ribosome formation (Mostafa et al., 1997). In this respect, Rady et
al. (2005) on amaryllis plants, Ebtsam Abdella et al. (2006) on Polianthes tuberosa, Hassanain et al. (2006) on
chamomile plants, Naguib et al. (2007) on Ruta graveolens and Yousef et al. (2010) on chamomile, they found
that foliar application with Fe, Zn or Mn individual or in mixtures affected positively on the above-mentioned
records of plant chemical composition.
Tables 3, 4, 5 and 6) clarified that gradual increase of GA3 followed by gradual increase in N, P, K, Fe, Zn,
Mn and total carbohydrates contents in leaves and flowers, with significant differences between mean values.
Likely values of total chlorophyll in leaves and carotenoids in flowers were significantly the highest when the
plants were treated with GA3 at 400 ppm. Increasing protein content by plant growth regulators may be due to
increase the formation of rough endoplasmic reticulum that provides the appropriate medium for increasing
polyribosome and mRNA (Kaber, 1987). These results are in agreement with, Ebtsam Abdella and Rady (2005)
on Asparagus sprengeri, Rawia et al. (2006) on croton plants, Shah and Samiullah (2007) on black cumin,
Azzaz et al. (2007) on Calendula officinalis, Faten et al. (2008) on Hemerocallis auantiaca, Kazaz, and
Karaguzel (2010) on Solidags * Hybrida and Gehan and Mona (2011) on Palanites aegyptiaca plants. They
concluded that pronounced enhancing influences on plant chemical composition was obvious as a consequence
of spraying GA3 at any concentration.
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J. Appl. Sci. Res., 9(1): 735-742, 2013
Table 3: Response of Calendula officinali L. plants to gibberellic acid and mixture of some micronutrients application on some
macronutrients contents in leaves and flowers during first season (2010\2011).
M.N. ppm
N%
P%
K%
In leaves
GA3 ppm
0.0
100
200
400
Mean
0.0
100
200
400
Mean
0.0
100
200
400
0.0
2.04 2.52 2.61 2.58
2.44
0.27 0.33 0.34 0.35
0.32
2.32 2.35 2.39 2.39
100
2.64 2.74 2.80 2.76
2.74
0.40 0.40 0.40 0.42
0.41
2.53 2.59 2.72 2.65
200
2.78 2.89 2.85 2.84
2.84
0.42 0.46 0.44 0.47
0.45
2.63 2.65 2.78 2.79
400
2.91 2.86 2.92 2.97
2.92
0.42 0.45 0.45 0.49
0.45
2.65 2.70 2.85 2.93
Mean
2.59 2.75 2.80 2.79
0.38 0.41 0.41 0.43
7.61 7.72 8.06 8.07
L.S.D.5%
M.N.
0.15
0.03
0.07
GA3
0.07
0.02
0.03
0.17
0.04
0.08
M.N.*GA3
In flowers
0.0
1.77 1.82 2.28 2.22
2.02
0.27 0.29 0.29 0.35
0.30
6.94 7.05 7.25 7.27
100
1.93 1.83 2.27 2.47
2.13
0.33 0.37 0.35 0.39
0.36
7.56 8.10 8.20 8.12
200
2.00 2.07 2.34 2.58
2.25
0.34 0.37 0.38 0.42
0.38
7.84 8.22 8.43 8.20
400
2.09 2.15 2.29 2.63
2.49
0.35 0.40 0.44 0.47
0.42
7.97 8.35 8.51 8.60
Mean
1.95 1.97 2.30 2.48
0.32 0.36 0.37 0.41
2.53 2.64 2.70 2.68
L.S.D.5%
M.N.
0.12
0.02
0.06
GA3
0.06
0.01
0.03
M.N.*GA3
0.14
0.02
0.07
M.N.= mixture of micronutrient (Mn+ Zn+ Fe) , GA3 = Gibberellic acid
Mean
7.09
7.87
8.14
8.35
2.38
2.67
2.72
2.79
Table 4: Response of Calendula officinalis L. plants to gibberellic acid and mixture of some micronutrients application on some
micronutrients contents during the first season (2010\2011).
M.N. ppm
Zn ppm
Mn ppm
Fe ppm
In leaves
GA3 ppm
0.0
100
200
400
Mean
0.0
100
200
400
Mean
0.0
100
200
400
Mean
0.0
0.13 0.14 0.14 0.17
0.19
0.17 0.20 0.19 0.20
0.15
7.55 7.79 7.53 7.68
7.64
100
0.18 0.21 0.22 0.24
0.25
0.23 0.24 0.24 0.27
0.21
8.06 8.20 8.34 8.49
8.27
200
0.19 0.22 0.24 0.25
0.25
0.23 0.25 0.26 0.27
0.23
8.44 8.59 8.70 8.55
8.57
400
0.20 0.23 0.24 0.26
0.28
0.25 0.27 0.29 0.31
0.23
8.51 8.59 8.84 8.67
8.65
Mean
0.22 0.24 0.25 0.26
0.18 0.20 0.21 0.23
8.14 8.29 8.35 8.35
L.S.D.5%
M.N.
0.02
0.02
0.14
GA3
0.01
0.01
0.07
0.02
0.02
0.16
M.N.*GA3
In flowers
0.0
0.11 0.12 0.16 0.16
0.14
0.18 0.19 0.21 0.19
0.19
7.09 7.27 7.17 7.22
7.20
100
0.16 0.17 0.22 0.24
0.20
0.23 0.24 0.25 0.27
0.25
7.98 8.10 8.28 8.42
8.20
200
0.16 0.19 0.23 0.25
0.21
0.25 0.26 0.28 0.26
0.26
8.29 8.28 8.41 8.65
8.41
400
0.18 0.21 0.25 0.25
0.22
0.26 0.29 0.29 0.29
0.28
8.20 8.40 8.68 8.69
8.49
Mean
0.15 0.17 0.22 0.23
0.23 0.25 0.26 0.25
7.89 8.01 8.14 8.25
L.S.D.5%
M.N.
0.02
0.02
0.11
GA3
0.01
0.01
0.06
M.N.*GA3
0.02
0.03
0.13
M.N.= mixture of micronutrient (Mn+ Zn+ Fe) , GA3 = Gibberellic acid
Table 5: Response of Calendula officinalis L. plants to gibberellic acid and mixture of some micronutrients application on leaves and flower pigments contents during the first season
(2010\2011).
M.N. ppm
Total carotenoides in leaves (mg\100g)
Total carotenoides in flowers
Total chlorophyll (mg\100g)
0.0
100
200
400
Mean
0.0
100
200
400
Mean
0.0
100
200
400
Mean
GA3 ppm
0.0
74.83
81.00
80.83
81.17
79.46
22.65
23.38
22.58
23.05
22.92
51.05
56.35
53.67
56.18
54.31
100
79.33
83.83
83.67
87.17
83.50
24.18
24.59
25.03
25.48
24.82
50.40
55.54
55.49
59.41
55.21
200
79.00
86.33
90.67
92.33
87.08
25.32
25.78
26.11
25.64
25.71
50.45
54.83
57.60
60.06
55.74
400
81.33
86.83
89.83
92.83
87.71
25.54
25.78
26.51
26.02
25.96
50.91
56.97
56.80
61.94
56.66
Mean
78.62
84.50
86.25
88.38
24.42
24.88
25.06
25.05
50.70
55.92
55.89
59.40
L.S.D.5%
M.N.
2.15
0.41
0.87
GA3
1.07
0.21
1.73
2.48
0.47
2.00
M.N.*GA3
M.N.= mixture of micronutrient (Mn+ Zn+ Fe) , GA3 = Gibberellic acid
Concerning the interaction between foliar application of GA3 and micronutrients mixture, recorded
determinations fluctuated significantly, however, observation of the highest ones differed as the parameter. The
highest content of Fe and total carotenoids in leaves were observed in association with GA3 at 400 and 200 ppm
of the mixture of micronutrients. However, the highest records of the other recorded parameters in both leaves
and flowers were observed resulted in spraying the plants with GA3 at 400 ppm concentration combined with
foliar application of micronutrients mixture at 400 ppm.
740
J. Appl. Sci. Res., 9(1): 735-742, 2013
Table 6: Response of Calendula officinalis L. plants to gibberellic acid and mixture of some micronutrients application on total
carbohydrate % during the first season (2010/2011).
M.N. ppm
In leaves
In flowers
0.0
100
200
400
Mean
0.0
100
200
400
GA3 ppm
0.0
12.40
13.94
14.47
14.76
13.89
9.51
10.59
11.55
12.16
100
13.50
14.58
14.96
15.87
14.73
10.18
11.73
12.40
13.30
200
14.06
13.71
14.93
16.01
14.68
10.56
12.31
12.72
13.91
400
14.32
14.90
15.11
16.16
15.12
10.65
12.40
13.39
14.12
Mean
13.57
14.28
14.87
15.70
10.38
11.76
12.52
13.37
L.S.D.5%
M.N.
0.79
0.47
GA3
0.39
0.24
0.91
0.55
M.N.*GA3
Mean
10.95
12.06
12.38
12.64
4. Anatomical study:
Data presented in Table (7) and illustrated in Figure (1), indicated that the marigold stem section diameter
was generally increased in response to foliar application of the different concentrations of gibberellic acid and
the mixture of micronutrients as compared to untreated plants. The highest increase was record by the
interaction between gibberellic acid at 400 ppm and the mixture of micronutrients at 200 ppm, this increase in
section diameter was mainly due to the increase in number of vascular bundle (which accompanied with the
increase in its length and width and the average diameter of xylem vessels) and the average diameter of pith
zone. These results agreed with those of Atya (2001) on soybean, Agamy et al. (2004) and Azzaz et al. (2007)
on marigold plants. These results indicated that, GA3 stimulated cell elongation and longitudinal cell diameter.
Table 7: Anatomical parameters of Calendula officinalis stem as affected by interaction between GA3 and mixture of micronutrients
Characters
Section
Cortex
No.
Av. length
Av. Number of
Av.
Av.
diameter
thickness µ
vascular
of vascular
xylem
diameter
diameter
Treatments
µ
bundle
bundle
vessels/Arch
of Mx
of pith
µ
vessels µ
µ
0ppm GA3
0ppm MN
3312
175
23
260
4
15
2325
100 ppm
4250
150
26
300
5
20
3190
MN
200 ppm
4125
175
22
2320
5.8
30
2875
MN
400 ppm
3875
200
29
260
5.5
25
2800
MN
100ppm GA3
0 ppm MN
3537
150
21
260
5.2
20
2800
100 ppm
4375
125
24
260
5
20
3800
MN
200 ppm
4000
200
20
300
6.1
30
3250
MN
400 ppm
3812
150
26
260
5
20
2800
MN
200ppm GA3
0 ppm MN
3500
125
24
270
5
20
2825
100 ppm
4250
200
22
330
6
20
2850
MN
200 ppm
4125
175
21
260
5
20
2625
MN
400 ppm
3925
150
27
290
5
15
2875
MN
400ppm GA3
0 ppm MN
3425
150
27
280
6
25
2875
100 ppm
4125
175
29
265
5
25
3312
MN
200 ppm
4600
150
25
350
6
30
3512
MN
400 ppm
3662
150
16
275
6
25
2625
MN
(MN) at the second season
741
J. Appl. Sci. Res., 9(1): 735-742, 2013
(x 30)
Fig. 1: Transections of calendula stem treated with:
a- 0ppm GA3+ 0ppm mixture of micronutrients (mn)
b-0ppm GA3+ 100ppm mixture of mn
d- 0ppm GA3+ 400ppm mixture of mn.
f- 100ppm GA3+ 100ppm mixture of mn
h - 100ppm GA3+ 400ppm mixture of mn.
j- 200ppm GA3+ 100ppm mixture of mn
l - 200ppm GA3+ 400ppm mixture of mn.
n- 400ppm GA3+ 100ppm mixture of mn
p- 400ppm GA3+ 400ppm mixture of mn.
(cx = cortex, p = pith and vb= vascular bundle).
c- 0ppm GA3+ 200ppm mixture of mn
e- 100ppm GA3+ 0ppm mixture of mn
g- 100ppm GA3+ 200ppm mixture of mn
I- - 200ppm GA3+ 0ppm mixture of mn
k- 200ppm GA3+ 200ppm mixture of mn
m - 400ppm GA3+ 0ppm mixture of mn
o- 400ppm GA3+ 200ppm mixture of mn
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