Int. J. Pure Appl. Sci. Technol., 17(2) (2013), pp.

79-89
International Journal of Pure and Applied Sciences and Technology
ISSN 2229 - 6107
Available online at www.ijopaasat.in

Research Paper
Effect of Foliar Spray of Ascorbic Acid, Zn, Seaweed
Extracts (Sea) Force and Biofertilizers (EM-1) on
Vegetative Growth and Root Growth of Olive (Olea
Europaea L.) Transplants cv. HojBlanca
Zulaikha R. Ibrahim

University of Duhok, Faculty of Agriculture and Forestry, School of Plant Production, Department of
Horticulture, Duhok, Iraq

Corresponding author, e-mail: (zulaukhabarwary@yahoo.com)

(Received: 8-5-13; Accepted: 12-6-13)

Abstract: The study was conducted at the nursery of Malta station/ Duhok.
Kurdistan region-Iraq, during the season 2011 to study the effect of Ascorbic
acid(Vitamin C), Zn, sea weed extracts(Sea force) and Biofertilizers (EM-1) on
vegetative growth and root growth of two year old of olive cultivar
(HojBlanca), the transplant were sprayed with four levels of Ascorbic acid (0,
250,500 and 750mg.l-1) by using Vitamin C (20 %)as a source of Ascorbic acid
and three levels of Zn(0, 15 and 30mg.l-1),three level of sea weed extracts( Sea
force) (0,1 and 2 mg.l-1)and three level of biofertilizer (EM-1) (0, 0.5 and 1
mL.L-1) Foliar application of Sea force at 2ml.l-1 increased significantly plant
height, leaf number and leaf dry weight. Increasing Zn level up 30mg.l-1
significantly increase stem diameter(mm).However 500mg.l-1 Ascorbic acid
increase significantly leaf area and shoot number/plant also Ascorbic acid at
750mg.l-1significantly increase total chlorophyll content and root dry weight.
Foliar application of (EM-1) at 0.5ml.l-1 increased significantly average shoot
length also (EM-1) at 1ml.l-1 significantly increase shoot dry weight and root
length.

Keywords: Foliar Application, Ascorbic acid, Zn, Sea force, EM-1, Olive
Transplants.
Int. J. Pure Appl. Sci. Technol., 17(2) (2013), 79-89 80

1. Introduction:
Olive belongs to the botanical order, Ligustrales, family Oleaceae; this family includes 30 genus
including Olea and has 600 species. Olive is botanically called (Olea eurovaea L.). Commercial
olives belong to the Eurovaea species, this species has two subspecies: oleaster and sativa (Bartolucci
and Dhakal, 1999).

Europaea is one of about 20 species of Olea found in subtropical regions of the world. Only Olea
europaea L. produces edible fruit. The Mediterranean region is native habitat (Sibbett et al., 2005).

In Iraq, olive trees growing in some areas of central and northern of Iraq, Nineveh is the governorate
leading olive producer, its cultivation in Nineveh, Kirkuk, Baghdad, Arbil and Duhok (Mahdi, 2007).

The importance of olive fruit is due to heavy loading and dietetic value, as the fruit is a good source
of vitamins (A, B, C, D, E, and K) and mineral like K, Ca, Mg and P (Ibrahim and Khlaef, 2007).(sh)
In addition, olive oil is filled with mono-unsaturated fatty acids and has many anti- oxidative
properties as phenolic acid (Hill and Giacosa, 1992; Trichopoulou, 1995; Stark and Madar, 2002).

Nowadays, there is a widespread use of antioxidants especially ascorbic acid for enhancing the
growth and productivity of fruit trees as well as controlling the incidence of most fruit disorders
(Ahmed et al., 1997b; Prestamo and Arroyo, 1999). Ascorbate is a major metabolite in the plant. This
antioxidant and its association with other components of the antioxidant system, protect the plant
against oxidative damage resulting from aerobic metabolism, photosynthesis and range of pollutants
(Nicholas, 1996). Ascorbic acid is also a cofactor for some hydroxylase enzymes (e.g. hydroxylase)
and violaxun via depoxidase. Ascorbic has also been implicated in regulation of cell division by
influencing progressing from G1 to S phase of the cell cycle. At the same time, Ascorbic acid is safe
to human, animal and environmental (Elade, 1992). Its application of it partially reduced the
application of hazard fungicide (Farag, 1996 and Ahmed et al., 1997b).

Aliniaeifard et al. (2008) studied the influence of antioxidants as application fertilizer on

photosynthetic characteristics and vegetative growth of one year old Olive (Olea Europal cv. Zard).
The results indicated that the application of ascorbic acid had a great effect on photosynthesis;
ascorbic acid caused an increase in stomata conductance, respiration, chlorophyll, and the leaf area
and leaf weight.

Nerway (2011) studied the effect of foliar spraying of ascorbic acid (0, 75 and 150mg. L-1) on
vegetative growth of grape cv. Rash-Mew (Vitis vinifera L.) under non-irrigated conditions. The
results showed that the foliar spraying of ascorbic acid at 75 and150mg. L.-1, had significantly
increased vegetative growth parameters (leaf area, leaf dry matter and total chlorophyll). Al-Douri
(2012) studied the response of pomegranate trees (Punica granatum L.) cv. Salimi to organic
fertilizer, NPK and Foliar spray of boron and ascorbic acid (0, 250, and 500mg As. L-1) during 2010
and 2011 seasons. The results indicated that the ascorbic acid spray caused a significant increment of
chlorophyll concentration of leaves in both seasons, Wassel et al. (2007) investigated the effect of
different concentrations of micronutrients, gibberelic acid and ascorbic acid (0, 500 and 1000 mg As.
L-1) on the growth of fifteen years old white Banaty seedless grapevine during 2000 and 2001
season. The results indicated to improving leaf area due to spraying with 500 and 1000mg. L-1
ascorbic acid.

Zinc is an essential component of various enzyme systems for energy production, protein synthesis,
and growth regulation. Zinc-deficient plants also exhibit delayed maturity. Zinc is not mobile in
plants so zinc-deficiency symptoms occur mainly in new growth. Poor mobility in plants suggests the
need for a constant supply of available zinc for optimum growth (Mortvedt, 1994). El-Mansi et al.
(1990) reported that Zn application increased all growth parameters of pea plants. El-Shazly and Dris
Int. J. Pure Appl. Sci. Technol., 17(2) (2013), 79-89 81

(2004) studied that the ′Anna′ apple trees when foliar sprays of cheated iron, manganese and zinc
alone or in a combination increased vegetative growth (shoot length and leaf area) as compared with
the control Farahat et al. (2007) studied that application of ascorbic acid 40 ppm and Zinc 40 ppm
separately promoted all the mentioned characters Hafez and El-Metwally (2007) studied that the
effect of Zn, K and Zn + K spray on leaf mineral content showed that leaf area significantly increased
as compared with the control.

Seaweed extracts is due to the antimicrobial activity of seaweeds against bacteria, yeast, and moulds
whereas the increased plant growth, yield and quality is resulted from the influence of these extracts
on cell metabolism via the induction of the synthesis of antioxidant molecules which could improve
plant growth and plant resistance to stress (Cardozo et al., 2007; Zhang and Schmidt, 2000). These
reports reveal that organic compounds rather than mineral elements are responsible for the effects.

Sea force contains almost every mineral and trace elements necessary for human and plant existence;
it also contains amino acid, and vitamins. Sea force is also a principle source of iodine (melson),
(SAFA Biological sea plant) (Berlyn and Russo, 1990). Sea force act as plant growth stimulants, their
effectiveness may be influenced by the species included and the manufacturing technique used
(Turan and Kose 2004). Mansour et al. (2006) investigated the impact of algae extract application to
thirty of 12 year old Anna Apple trees. Results showed that the applied of algae extract was very
effective in stimulating the shoot length, leaf area, total leaf carbohydrates and leaf mineral content.

Chouliaras et al (2009) recommended the combination of NH4No3 + borax + sea force to trees, in
order to improve their nutrition status.

Francesco et al. (2010) investigated the influence of the bio-stimulant (Sea force) on the vegetative
and productive performance of strawberry. The results showed that the vegetative growth and leaf
chlorophyll content increased significantly as compared to control.

Bio fertilizers play a very important role in improving soil fertility by fixing atmospheric nitrogen,
both in association with plant roots and without it, solubilise insoluble soil phosphates and produce
plant growth substances in the soil. They are in fact being promoted to harvest the naturally available
biological system of nutrient mobilization (Venkatashwarlu, 2008).

Fawzia (2003) showed that the soil and /or foliar spraying of bio stimulants EM, to Kelsey plum
during the 2001 and 2002 season resulted in the greatest leaf area and chlorophyll reading, the least
number of leaves /shoot and increment in the stem length.

The aims of this experiment are to study the effects of Ascorbic acid, Zn, sea weed extracts (sea
force) and Biofertilizers (EM-1), on vegetative growth and root growth of olive, and chlorophyll
contents of olive transplants cv. (HojBlanca) which newly entered to the region.

2. Materials and Methods

The study was carried out during 2011 in the nursery of Malta station/ Duhok. Kurdistan region-Iraq.
Uniform and healthy olive cv (HojBlanca) transplant of (2) years old. The experiments were started
in April 21th 2011, as transplants were grown in pots each of (5kg) weight, filled with river loamy
soil Restrepo-Diaz et al. (2008),with four levels of Ascorbic acid (0, 250,500 and 750mg.l-1), three
levels of Zn(0, 15 and 30Zn.mg.l-1),three level of Sea force (0,1 and 2 mg.l-1) and three level of (EM-
1) (0, 0.5 and 1 mL.L-1) were sprayed at 21of April and repeated at same concentrations in 21 of
May. Agha and Daoud (1991).
Int. J. Pure Appl. Sci. Technol., 17(2) (2013), 79-89 82

Soil
Taken (5kg) from soil studied and analyzed which used in the experiment, they were air dried,
grounded and passed through a 2mm sieve, to prepare them for soil physical and chemical analysis
according to Page, et al. (1982) and AOAC, (1995) methods at soil department laboratory,
Agriculture College, Duhok University (Table 1).

The transplants were foliar sprayed with solutions containing Ascorbic acid, Zn, Sea force and (EM-
1) to drip point (run off) using 2 litters hand sprayer at 21 of April, 2011. The transplants were left
under nursery conditions till end of the experiments.

The experiment was arranged as Factorial Randomized Complete Block Design

Table (1): Some physical and chemical properties of nursery soil

Soil properties Value

pH 7.1
Total nitrogen N% 0.07
Available Phosphorus 5.2
(ppm)
Exchangeable K+ meq/L 0.033
+2
Exchangeable Ca meq/L 1.2
CaCo3 meq/L 12.5
EC (ds.m) 0.20
Organic mater (%) 0.34
Sand% 62.05
Silt% 22.99
Clay% 14.96
Soil texture Loamy sand

(R.C.B.D) using four factors (4×3×3×3): Ascorbic acid, Zn, sea weed extracts (sea force) and
Biofertilizers (EM-1), levels consequently, with five replicate. Each replicate consisted of one
transplant. The data were analyzed statistically, and comparisons were made using Duncan's multiple
range test at 5% probability (Al-Rawi and Khalaf-Allah, 1980). All data were tabulated and
statistically analyzed with computer using (SAS, 2000).

The following measurements were recorded on 15th September 2011:

1- Plant height (cm)

2-Leaves number/plant
3-Stem diameter (mm)
4- Primary shoots length average (cm).
4- Single Leaf area cm2
6- Primary shoots number / plant. Lengths per transplant (cm)
7-Total Chlorophyll Content (%)
8- Shoot dry weight (%).
9- Leaf dry weight (%).
10-Root length (cm)
11-root dry weight (%)
Int. J. Pure Appl. Sci. Technol., 17(2) (2013), 79-89 83

3. Results and Discussion:

1-Plant height (cm): The obtained results of (Table 2) revealed that (sea force) was the most
effective rate at (2ml.L-1) level, as it gave the highest plant height as compared to control.

The lowest plant height (36.00) was recorded on the untreated (control) trees.
No significant difference was noticed when the transplants were treated with Zn at (30mg.l-1) and (sea
force) at (2ml.L-1) level.

2- Stem diameter (mm): Foliar spray with Zn at (30mg.l-1) significantly increased stem diameter as
compared with the untreated transplants. The highest stem diameter (5.66mm) was recorded in
(30mg.l-) Zn.

3- Leaf numbers/plant: Data in figure (2) showed that (sea force) at (2ml.L-1) level gave significant
increase in leaf number compared to the control.

4- Leaf area cm2: Table (2) showed that significant differences in leaf area due to the treatments.
Ascorbic acid (500mg.l-1) treatment significantly increased leaf area (7.99 cm2) compared to the
untreated transplants.

5- Leaf dry weight: Leaf dry weight increased significantly and gradually by foliar spray with (sea
force) at (1ml.L-1) level (54.46) whereas the lowest leaf dry weight (43.60) was recorded at foliar
spray with Zn at (30mg.l-1).

Table (2): Effect of foliar spray of Ascorbic acid, Zn, seaweed extracts (Sea f orce) and Biofertilizers
(EM-1) on some vegetative growth characteristic of olive (Olea europaea L.) transplants cv

Parameters
Plant Stem Leave Leaf Leaf dry
Treatments
height(cm) diameter(mm) number area(cm2) weight(%)
36.00 3.33 85.00 5.06 48.66
Control
g d G g bc
Ascorbic 47.88
42.67 4.33 115.00 5.99
acid 250 b-d
f b-d E de
mg.L-1
Ascorbic 48.66
62.33 4.33 146.66 7.99
acid 500 bc
c b-d C a
mg.L-1
Ascorbic 44.22
80.00 4.66 122.33 6.83
acid 750 cd
b a-c D bc
mg.L-1
57.33 4.66 116.33 5.85 52.37
Zn 15 mg.l-1
de a-c E e ab
43.60
5.66 123.00 5.44
Zn 30 mg.l-1 89.66 d
A D f
a
Sea force 53.66 5.33 178.66 6.20 54.46
1ml.L-1 e ab B d a
Sea force 90.33 5.33 220.66 7.02 44.98
2ml.L-1 a ab A b cd
48.51
EM-1 0.5 4.66 106.33 6.54
60.33 b-d
mL.L-1 a-c F c
cd
Int. J. Pure Appl. Sci. Technol., 17(2) (2013), 79-89 84

4.00 50.59
EM-1 1 62.66 124.00 6.86
cd ab
mL.L-1 c D b

Means within a column, following with the same latter are not significantly different according to
Duncan multiple range test at the probability of 0.05 levels.

6- Average shoots length (cm): Table (3) shows that the highest average shoots length (51.00) was
recorded on transplants treated with (EM-1) at (0.5mL.L-1) (EM-1) at (1mL.L-1) levels had no
significant influence on average shoots length compared with the control.

7- Shoots number /plant: The highest significant shoot number was obtained from transplant foliar
sprayed with Ascorbic acid (500mg.l-1) (9.66).

Whereas foliar spray with (EM-1) levels had no significant influence on shoot number of transplants
compared with other treatments.

8- Shoot dry weight: Table (3) reveal that transplants treated with (EM-1) at rate of (1 mL.L-1) had
significantly higher shoot dry weight as compared with the control.

Table (3): Effect of foliar spray of Ascorbic acid, Zn, seaweed extracts (Sea f orce) and Biofertilizers
(EM-1) on shoot growth characteristic of olive (Olea europaea L.) transplants cv

Parameters
Treatments
average shoot
Shoot number Shoot dry
length(cm)
weight(%)
Control 20.66 4.33 63.96
e e ab
Ascorbic acid 250 4.66
38.00
mg.L-1 de 58.31
cd
bc
Ascorbic acid 500 9.66
mg.L-1 35.00 a 53.61
d cd

Ascorbic acid 750 7.33 52.61

46.66
mg.L-1 b cd
ab
Zn 15 mg.l-1 4.33
36.00 e 51.92
d d

Zn 30 mg.l-1 5.66
44.66 c 50.35
b d

Sea force 1ml.L-1 5.33 51.67

41.66 cd d
bc
Sea force 2ml.L-1 35.33 7.66
d b
58.50
Int. J. Pure Appl. Sci. Technol., 17(2) (2013), 79-89 85

bc

EM-1 0.5 mL.L-1 3.33

51.00 f 50.80
a d

EM-1 1 mL.L-1 3.33 64.69

22.33
f a
e

Means within a column, following with the same latter are not significantly different according to
Duncan multiple range test at the probability of 0.05 levels.

9- Total Chlorophyll Content: Total chlorophyll percentage (measured by SPAD) in leaf of

transplant treated with 750 mg.l-1 Ascorbic acid significantly surpassed as compared with other
treatment.
No significant difference was noticed when the transplants were treated with Ascorbic acid at
(750,500 mg.l-1) and Zn at (30mg.l-1) (71.63), (71.60) and (71.03) respectively.

Whereas the lowest Chlorophyll Content (58.70) was recorded on the untreated (control) transplant.

10-Root length (cm): Table (3) reveal that transplants treated with (EM-1) at rate of (1 mL.L-1) had
significantly higher root length as compared with the other treatment.

Whereas foliar spray with Ascorbic acid at (750mg.l-1) had no significant influence on shoot number
of transplants.

11-root dry weight (%): Data in figure (4) showed that Ascorbic acid at (750mg.l-1) level gave
significant increase in root dry weight (68.03) compared to other treatments.

Whereas the lowest root dry weight (27.83) was recorded at foliar spray with (EM-1) at rate of (1
mL.L-1).

Table (4): Effect of foliar spray of Ascorbic acid, Zn, seaweed extracts (Sea force) and Biofertilizers
(EM-1) on chlorophyll content and some root growth characteristic of olive (Olea europaea L.)
transplants cv

Parameters
Treatments Chlorophyll Root Roor dry
content (%) length(cm) weight(%)
Control 58.70 25.33 40.10
e ef f

Ascorbic acid 250 63.60 25.00 68.03

mg.L-1 cd ef a

Ascorbic acid 500 71.60 28.00 46.60

mg.L-1 a d d

Ascorbic acid 750 71.63 23.66 62.86

mg.L-1 a f b
Int. J. Pure Appl. Sci. Technol., 17(2) (2013), 79-89 86

Zn 15 mg.l-1 68.73 27.33 42.73

b d e

Zn 30 mg.l-1 71.03 25.00 53.96

a ef c

Sea force 1ml.L-1 63.13 36.66

d b 43.06
e
Sea force 2ml.L-1 62.93 26.33 41.93
d de e

EM-1 0.5 mL.L-1 67.93 31.33 35.66

b c g

EM-1 1 mL.L-1 64.26 41.66 27.83

c a h

Means within a column, following with the same latter are not significantly different according to
Duncan multiple range test at the probability of 0.05 levels.

Discussions
It was observed from table (2-4) that the foliar spraying of Ascorbic acid had a positive effect on the
single leaf area, total chlorophyll the vegetative growth parameters of the vine, so that the spraying
transplants with (500mg.l-1) significantly increased the single leaf area and total chlorophyll. These
results were supported by those of Ahmed and Morsy (2001); Wassel et al. (2007) and Fayed (2010).
These increases in the above parameters by using ascorbic acid may be due to the fact that Ascorbic
acid as an anti oxidant has an effect as plant growth regulators (Ahmed et al., 1997a; Johnson et al.,
1999) and its role in activating both cell division and elongation in meristematic tissues, as well as
the bio synthesis of organic foods (Nijjar, 1985).

Concerning the effect of Zn on increasing stem diameter may be due to the role of Zn as an essential
element for tryptophan synthesis, one of the auxin IAA precursor (Taiz and Zeiger, 2002). These
results are in agreement with those of Khattak et al., 2006; Mostafa and saleh, 2006 and Al-A‫ۥ‬arji and
Al-Hamadany, 2006 who reported that application of Zn increased vegetative growth and thickness of
introde, increased cell size and cell number which may be responsible for the increase of stem
diameter.

The data in tables (2 to 4) indicate that foliar spraying of sea force at (2ml.L-1) had significant effects
on vegetative growth parameter, as it gave the highest plant height, number of leaves, and Leaf dry
weight as compared to control. The reason behind increasing dry leaf weight could be attributed to
increasing petioles of N, P and K % in the leaves (Mancuso et al., 2006) and their role in activating
the cell division and increasing biosynthesis of organic products that lead to accumulation of
carbohydrates and protein in leaves. The results are the same found by Eman and Abd-Allah (2008);
Turan and Kose (2004); Macantsaoir and Archer (2008) and (Mac and Archer 2010).

The role of effective microorganism EM in agriculture has been reported as one of decomposing
organic matter, while enhancing the qualities of the rhizosphere. Thus crop yields have been
enhanced on a sustainable basis in organic systems, and environmental quality preserved (Parr et al.,
1997). The studies on the success of EM in crop production are many. Research on apples in Japan
Int. J. Pure Appl. Sci. Technol., 17(2) (2013), 79-89 87

(Fujita, 2000) and herbage grasses in Holland and Austria (Bruggenwert, 2001 and Hader, 2001)
illustrates this phenomenon very clearly.

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