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

Abstract

This 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 by using Vitamin C as a source of Ascorbic acid and three levels of Zn, three level of sea weed extracts (Sea force) and three level of biofertilizer (EM-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. 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 at 1ml.l-1 significantly increase shoot dry weight and root length.

Headnote

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.

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 (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).

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 leftunder nursery conditions till end of the experiments.

The experiment was arranged as Factorial Randomized Complete Block Design

(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 (%)

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).

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.

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).

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

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