Effect of Bio-phosphate and Chemical Phosphorus Fertilizer Accompanied with Foliar Application of Micronutrients on Yield, Quality and Phosphorus and Zinc Concentration of Maize

Abstract

Field experiment was conducted at Kerman Agricultural and Natural Resources Research Centre (Iran) during 2008-2009 to evaluate the effect of bio-fertilizer, phosphorus and foliar application of micronutrients on dry matter accumulation, yield, and phosphorus and zinc concentration of maize (Zea mays L.). A split plot experiment based on randomized complete blocks design (RCBD) with four replications was followed in the study. The micronutrients foliar application in two levels (foliar application and non foliar application) were the main plots, and four levels of phosphate (T1: 0 (no fertilizer), T2: 100 kg ha-1 P2O5, T3: 100g bio-phosphate, T4: 100g bio-phosphate with 50 kg ha-1 P2O5) as the sub plots. Results showed that micronutrients foliar application and biological and chemical phosphorus fertilizers had a significant influence on dry matter accumulation. The maximum dry matter accumulation was obtained by applying 50 kg/ha P2O5 plus bio-fertilizer. Grain yield, 1000-seed weight and protein content of grain were significantly affected by micronutrients and phosphorus fertilizers treatments. Micronutrients foliar application and phosphorus fertilizers interaction had no significantly effect on grain yield, 1000-seed weight and grain protein content. Grain phosphorus and zinc concentration where significantly increased by application of micronutrients and phosphorus fertilizers.

Full text

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www.ccsenet.org/jas Journal of Agricultural Science Vol. 3, No. 4; December 2011
ISSN 1916-9752 E-ISSN 1916-9760
22
Effect of Bio-phosphate and Chemical Phosphorus Fertilizer
Accompanied with Foliar Application of Micronutrients on Yield,
Quality and Phosphorus and Zinc Concentration of Maize

Mohammad Galavi, Khatoon Yosefi & Mahmod Ramrodi
Department of agronomy, collage of agriculture
University of Zabol, P.O.Box 98615-538, Zabol, Iran
Tell: 98-542-222-5077 E-mail: mgalavi@yahoo.com

Sayed Roholla Mousavi (Corresponding author)
Aligoudarz Branch, Islamic Azad University, Aligoudarz, Iran
Tell: 98-664-223-4731 E-mail: rr_mousavi@yahoo.com

Received: March 2, 2011 Accepted: March 23, 2011 Published: December 1, 2011
doi:10.5539/jas.v3n4p22 URL: http://dx.doi.org/10.5539/jas.v3n4p22

Abstract
Field experiment was conducted at Kerman Agricultural and Natural Resources Research Centre (Iran) during
2008-2009 to evaluate the effect of bio-fertilizer, phosphorus and foliar application of micronutrients on dry
matter accumulation, yield, and phosphorus and zinc concentration of maize (Zea mays L.). A split plot
experiment based on randomized complete blocks design (RCBD) with four replications was followed in the
study. The micronutrients foliar application in two levels (foliar application and non foliar application) were the
main plots, and four levels of phosphate (T1: 0 (no fertilizer), T2: 100 kg ha-1 P2O5, T3: 100g bio-phosphate, T4:
100g bio-phosphate with 50 kg ha-1 P2O5) as the sub plots. Results showed that micronutrients foliar application
and biological and chemical phosphorus fertilizers had a significant influence on dry matter accumulation. The
maximum dry matter accumulation was obtained by applying 50 kg/ha P2O5 plus bio-fertilizer. Grain yield,
1000-seed weight and protein content of grain were significantly affected by micronutrients and phosphorus
fertilizers treatments. Micronutrients foliar application and phosphorus fertilizers interaction had no significantly
effect on grain yield, 1000-seed weight and grain protein content. Grain phosphorus and zinc concentration
where significantly increased by application of micronutrients and phosphorus fertilizers.
Keywords: Bio-phosphate, Dry matter, Maize, Micronutrient, Yield
1. Introduction
Integration of chemical and bio-fertilizers is one of the ways to increase production in sustainable agriculture
(Ali et al. 2008; Sharma, 1999). Phosphorus bio-fertilizers bacteria such as Bacillus and Pseudomonas increased
soil soluble phosphorus by secreting organic acids and phosphatase enzyme (Ehteshami et al. 2007).
Environmental problems caused by irregular application of chemical fertilizers, inappropriate energy production
methods and excessive consumption costs have all had harmful effects on biological cycles and destroyed
farming stability systems; these factors altogether encourage the application of bio fertilizers (Kannayan, 2002).
Ability of micronutrients absorption (especially zinc) is very important for optimum growth of plants. In most of
the Iranian soils pH is high and they are also calcareous. In this type of soils solvability of micronutrients is less
and it decrease absorption of micronutrients by plant, finally requirement of plants increasing to this elements
(Mousavi et al. 2007). Maralin, (2009) showed that yield and Zn and Fe concentration of wheat increased by Zn
and Fe foliar application. Plant height, stem diameter and leaf area index of maize where significantly increased
by application of nitrogen and phosphorus fertilizer 8 weeks after sowing (Onasanya et al. 2009). Hamidi et al.
(2008) in a study showed that fresh weight, number of leaves above the ear, yield of silage forage and
vegetative growth of maize increased by application of PGPR (Plant growth promoting rhizobacteria).
Hassanzadeh et al. (2006) reported grain yield and dry matter production in barley increased by application of

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phosphate-solution bacteria and chemical phosphorus fertilizer. El-Gizawy and Mehasen, (2009) showed that
application of chemical phosphorus fertilizer with phosphate-solution bacteria had a significant effect on bean
grain yield, yield components, nitrogen content, and content of phosphorus and zinc in the grain. Maize growth
and dry weight increased by plant growth promoting rhizobacteria (PGPR) application (Zahir et al. 1998; Javed
et al. 1998). Application of bio-fertilizer with 50% of chemical nitrogen, phosphorus and potassium (NPK)
fertilizers increased maize vegetative growth, plant height, branch number, fresh and dry weight and total
carbohydrate on dry matter as compared to chemical fertilizers treatments (Mahfouze and Sharafeldin, 2007).
(Note 2)
With regards to importance of phosphate absorption from soil on the maize feeding the present research was
done in order to evaluate the effect of bio-phosphate and chemical fertilizers combined with micronutrients foliar
application on dry matter accumulation trend, growth and qualitative characteristics of maize (S.C.704). (Note 1)
2. Materials and methods
2.1 Description of the project site
This experiment was carried out during 2008-2009 at the Kerman Natural Resources and Agriculture Research
Center, Iran, located in 56º34´ longitude and 29º55´ latitude and, 2044m Altitude from sea level with an arid and
semi-arid climate. The pH of soil field experiment was 8 and soil texture was loamy, (physical and chemical
properties of soil in experimental field were presented in table 1). Experiment was conducted in split plot within
a randomized complete block design with four replications. The main plots included foliar application of
micronutrients containing Fe, Zn, Mn, and Cu elements together with control application (non foliar application),
and sub plot were considered four levels of phosphorus fertilizers: (T1: 0 (no fertilizer), T2: 100 kg ha-1 P2O5, T3:
100g bio-phosphate, T4: 100g bio-phosphate with 50 kg ha-1 P2O5)Sowing was done as rows in 75cm wide rows
with 20cm spacing within-rows with six rows per subplot by Single Cross 704 cultivar, (Single Cross 704 was
chosen because this cultivar had superiority relative to other cultivar in the last few years in experimental region).
Foliar application of micronutrients was done in 4 liters per thousand at stem extension and staminate
inflorescence emergence stages. Prior to planting, seeds were inoculated with biological phosphorus fertilizer
and chemical phosphorus fertilizer was utilized as strip takes under seed. All operations were done regularly
during the growing season.
2.2 Crop sampling and calculation
Dry matter was determined 35 days after sowing of one square meter area in each plot after eliminating the
marginal effect by oven at 700C for 48 hours, and was repeated once every 15 days until the end of the growing
period. 1000-seed weight and grain yield was measured at the end of growing season after of physiology
maturity by harvesting 5 m2 from each plot. Percent of protein was obtained by following formula after
determining amount of nitrogen in the laboratory by Kjeldahl method (Jones, 1991).
Pr = 6.25 × Nt × DM
Pr = Percent of protein
Nt = Percent of total N
DM = Percent of dry matter
Concentration of zinc and phosphorus was obtained by atomic absorption and spectrophotometer methods
respectively.
2.3 Statistical analysis
Data analysis was done by using SAS and MSTATC software. The ANOVA test was used to determine
significant (p≤0.01 or p≤0.05) treatment effect and Duncan Multiple Range Test to determine significant
difference between individual means.
3. Results and discussion
3.1 Dry matter accumulation
Analysis of variance showed that dry matter accumulation was significantly affected by phosphorus fertilizer
treatments during the stages of sampling (from 35 to 125 days after sowing); Also dry matter accumulation was
significantly affected by foliar application of micronutrients during the stages of sampling except the first stage
(35 days after sowing) (Table 6 and 2). Trend of dry matter accumulation showed that seed inoculation by
biological phosphorus fertilizer before sowing increased dry matter accumulation. Highest dry matter
accumulation (1810 g/m2) was obtained by application of biological fertilizer + 50 kg ha-1 P2O5 (Figure.

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1). Maize resistant to lodging, precocity and its quality and quantity increased by increasing phosphorus
availability; also grain yield increased by increasing weight and grain number per ear (Ehteshami et al. 2007).
Bio-phosphate bacteria efficiency increased by application of biological phosphorus fertilizer combined with
chemical phosphorus fertilizer. Pseudomonas bacteria in the bio-phosphate produced auxin and gibberellin
hormones and vitamins, therefore dry matter increasing could be attributed to the ability of bacteria
(Hassanzadeh et al. 2006). Effect of foliar application of micronutrients on dry matter accumulation appeared 50
days after sowing, the highest difference between foliar application of micronutrients and non foliar application
treatments on dry matter accumulation was observed 95 days after sowing (Figure. 2).
3.2 1000-Seed weight
1000-seed weight was significantly affected by biological and chemical phosphorus fertilizers, but foliar
application of micronutrients had no significant effect on 1000-seed weight (Table 3). Application of biological
fertilizer + 50 kg ha-1 P2O5 increased 1000-seed weight to 304.97g which was 9.77% higher as compared to
control. Maize 1000-seed weight was dependent on the ability of plants to provide food for the reservoir and
environmental conditions such as moisture and nutrients during grain filling (Andrade et al. 1999).
3.3 Grain yield
Analysis of variance showed that grain yield was affected by phosphorus fertilizers and foliar application of
micronutrients (Table 3). According to the results, highest and lowest yields were obtained by application of
biological fertilizer + 50 kg ha-1 P2O5 and control treatments respectively. Maize yield increased by application
of biological phosphate fertilizer, that it could be due to increasing other nutrient absorption, also biological
phosphate fertilizer can be used as a solution for increasing phosphate and micronutrients sorption in the alkaline
soil (Hassanzadeh et al. 2006). Tahir et al. (2009) reported that grain weight and yield of maize increased by
application of zinc. Increase of grain yield under the influence of phosphate fertilizers, biological fertilizer + 50
kg ha-1 P2O5, can be attributed to the ability of phosphate solution bacteria in bio-fertilizer in increasing available
phosphorus of insoluble phosphorus sources. In another study Rokhzadi et al. (2004) reported that grain yield of
chickpea increased by application of biological fertilizers.
3.4 Grain protein
Percent of grain protein was significantly affected by biological and chemical phosphorus fertilizers, but foliar
application of micronutrients had no significant effect on grain protein (Table 3). Mean comparisons showed that
maximum (9.74%) and minimum (8.22%) grain protein was obtained by application of biological fertilizer + 50
kg ha-1 P2O5 and control respectively (Table 4). Phosphate solubilizing microorganisms increased available
phosphorus and nitrogen in the soil which increases their concentration by plant, also increased the grain protein
(Mehrvarz et al. 2008). Hemati (2005) reported wheat protein increased by application of micronutrients such as
Zn, Mn and Fe.
3.5 Grain phosphorus concentration
Result showed that grain phosphorus concentration was significantly affected by biological and chemical
phosphorus fertilizers and foliar application of micronutrients, also phosphorus fertilizers and foliar application
interaction had a significant effect on percent of grain phosphorus (Table 3). Maximum phosphorous content
(0.24%) was obtained by application of biological fertilizer + 50 kg ha-1 P2O5 and foliar application of
micronutrients which, was 25% higher as compared to control treatment (no fertilizer) (Table 5). Phosphorus
uptake by plant decreased by increasing Zn in the soil, but Zn application by foliar application increased
phosphorus absorption of soil by plants (Bukvic et al. 2003). Phosphate-solubilizing microorganisms had
important role in phosphorus solubility and uptake. The bacteria released phosphorus from organic and inorganic
soils during the mineralization process also increased the absorption of phosphate from phosphate rock, and save
a reservoir of phosphorus in the presence of carbon unstable by mineralization process (Alikhan et al. 2009).
3.6 Grain zinc concentration
Phosphorous fertilizers and foliar application of micronutrients interaction had a significant effect on grain zinc
concentration (Table 3). Mean comparisons showed that maximum zinc concentration (16.15 mg/kg) was
obtained by application of biological fertilizer + 50 kg ha-1 P2O5 and foliar application of micronutrients; also
minimum zinc concentration (14.95 mg/kg) was obtained by control. There wasn’t significant difference
between biological, chemical, biological fertilizer and 50 kg ha-1 P2O5, foliar and non foliar application on zinc
concentration (Table 5). Foliar application is very fast method for providing requires elements in plants because
nutrients are uptake quickly as compare to uptake that through plant roots. Zinc transferred to grain by
remobilization and it remobilization was higher than other micronutrients (Ranjbar and Bahmaniar, 2007;

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Kochian, 1991). The concentration of zinc in wheat increased to three times by Zn and Fe foliar application in
the tillering and heading stage (Maralian, 2009). Zinc absorption reduced by phosphorus application in the soil,
but biological phosphorus fertilizers due to having phosphate solubilizing bacteria had no interfere on other
elements uptake (Salimpour et al. 2010). Zakaria et al. (1997) reported phosphorus and zinc uptake by cotton
increased by application of biological phosphorus fertilizers and zinc foliar application.
4. Conclusion
Application of phosphorus fertilizers combined with foliar application of micronutrients increased dry matter
accumulation. Biological phosphorus fertilizers efficiency increases if combined with 50% chemical phosphorus
fertilizers. Integrating biological and chemical phosphorus fertilizers with foliar application of micronutrients
increased grain yield as compared to control. Biological phosphorus fertilizer with 50kg chemical phosphorus
fertilizer combined with foliar application of micronutrients increased grain zinc and phosphorous concentration.
Zinc was available through foliar application for plant, thus application of phosphorus fertilizers had no negative
effect on zinc uptake by plant in the soil, thereupon zinc and phosphorus concentration increased in the grain.
Application of biological fertilizer was significantly increased 1000-seed weight and percent of grain protein,
because nutrients absorption capability and plant absorption balance increased by phosphate-solubilizing
microorganisms. Grain yield and quality increased by foliar application of micronutrients due to increasing
elements transfer from leaves to grain. Application of chemical phosphorus fertilizer decreased to 50% by
integrating biological and chemical phosphorus fertilizers without yield loss. Also environmental pollution was
reduced by decreasing consumption of chemical fertilizers. Overall application of biological phosphorus
fertilizers with chemical phosphate fertilizer in addition to yield increasing could be a strategy to achieve
sustainable agriculture.
References
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management practices for yield improvement. Australian Journal of Crop Science, 2(3), 150-157.
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application of plant growth promoting rhizobacteria (PGRP) on the Yield of fodder maize (Zea mays L.).
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Hassanzadeh, E., Mazaheri, D., Chaichi, M. R., & Khavazi, K. (2006). Efficiency of phosphorus solubilizing
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Hemati, A. (2005). Application of soil and spray application of iron, zinc and manganese on yield and bean
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Kannayan, S. (2002). Biofertilizers for sustainable crop production, Biothecnology of biofertilizers. Narosa
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quality and enrichment on potato. Asian Journal
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yield response of maize (Zea mays L.) to different rates of nitrogen and phosphorus fertilizers in Southern
nigeria. World Journal of Agriculture Science, 5, 400-407.
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growth and yield of maize (Zea mays L.). Soil and Environmental Journal, 28(2), 179-183.
Zahir, A. Z., Arshad, M., & Khalid, A. (1998). Improving maize yield by inoculation with plant growth
promoting rhizobacteria. Pakistan Journal of Soil Science, 15, 7-11.
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Notes
Note 1. Single Cross 704 (S.C.704)
Note 2. Nitrogen, phosphorus and potassium (NPK)

of Plant Sciences, 6(8), 1256-1260.
Table 1. Soil analysis result for physical and chemical characteristics
Soil
texture depth
(cm)
Cu Mn Fe Zn K P pHEC
(dS/m-1)
OC
(%)
Soil
Characteristic
(ppm)
0.788.1 6.02 0.42 200 6.58 0.52 0.82 loamy 0-30 Value

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Table 2. ANOVA of the effects of biological and chemical phosphorus fertilizers with foliar application of
micronutrients on dry matter accumulation (g/m2)

SOV df
35 1 50
65
Replication 3 18.90
39.25
148.33
Micronutrients
foliar
application
Error 3 0.71
112.26 8119.81
Phosphorous
fertilizers 3 358.20** 2900.50** 55788.80**
Phosphorous
fertilizers×
Micronutrients
foliar
application
Error 18 10.44
149.76
2256.94
CV (%) 7.27
10.37
9.43
1- Days after sowing
2- ns= Non significant, ** = p < 0.01 and * = p < 0.05

Table 3. ANOVA of the effects of biological and chemical phosphorus fertilizers with foliar application of
micronutrients on yield and quality characteristics of maize

SOV df
Grain yield
(ton/ha-1)
Replication 30.17
Micronutrients foliar application 1 2.39*1
Error 30.62
Phosphorous fertilizers 313.67**
Phosphorous fertilizers× Micronutrients
foliar application
3 0.15 ns
Error 180.35
CV (%) 7.97
1- ns= Non significant, ** = p < 0.01 and * = p < 0.05
MS
80

95
110
2937.11

97887.60**
125
13545.44

6308.20**
2894.97

140495.10**
2975.23

49905.90**

1

1.00 ns2

5021.02**

4328.90**
1606.92

79283.80**


4757.80 ns
17427.54

147601.30**


3747.63 ns
9978.76

162077.40**


2283.25 ns
12050.45

137026.60**


4361.60 ns


3


4.53 ns


327.45 ns


1135.28 ns
5469.04
8.38
1946.44
8.49
9325.73 9718.57
6.22 6.79
MS
1000-seed weight
(g)
53.58
155.01ns
5.14
1146.10**

15.75 ns
35.63
2.04
Protein
(%)
0.48
0.23
0.18
3.47**

0.13 ns
0.16
4.50
P
(%)
0.00005
0.0004**
Zn
(mg/kg)
0.14
1.50**
0.0000006
0.001**

0.0001*
0.00002
2.04
0.10
0.90**

0.40*

0.10
2.05

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Table 4. Means comparison of effects of biological and chemical phosphorus fertilizers with foliar application of
micronutrients on yield and quality characteristics of maize
Treat
Grain yield
(ton/ha)
7.79a
1000-seed weight
(g)
294.52a
Protein
(%)
9.06a 0.24b 15.76a
P
(%)
Zn
(mg/kg)
Micronutrients
Micronutrients foliar application
Non micronutrients foliar application
Chemical P
Biological P × Chemical P
7.23a
8.09b
8.81a
290.12a
298.61b
304.97a
8.89b 0.23b 15.33b
9.24b 0.24b 15.64a
9.74a 0.25a 15.86a
Fertilizer
Biological P
No fertilizer
7.32c
5.75d
287.90c
277.81d
8.69c 0.24b 15.62a
8.22d 0.22c 15.07b
1- Columns means followed by the same letter are not significantly different at 0.05 or 0.01 probability level

Table 5. Effects of biological and chemical phosphorus fertilizers and foliar application of micronutrients
interaction on grain phosphorus and zinc concentration
Treatments

Micronutrients foliar application Biological P × Chemical P
P (%)
0.242b1
0.257a
Zn (mg/kg)
15.99ab
16.15a
Chemical P



Biological P
No fertilizer
Chemical P
0.242b
0.220d
0.234c
15.97ab
15.19cd
15.29cd
Non micronutrients foliar applicationBiological P × Chemical P
Biological P
No fertilizer
0.242b
0.232c
0.223d
15.57bc
15.27cd
14.95d
0.46


LSD
0.007
1- Columns means followed by the same letter are not significantly different at 0.05 or 0.01 probability level

Table 6. Means comparison of the effects of biological and chemical phosphorus fertilizers with foliar
application of micronutrients on dry matter accumulation (g/m2)
Days after sowing
35 50
65
80
95
110
125
Micronutrients
Micronutrients foliar application 44.57a1130.52a540.10a928.73a1711.64a 1607.32a 1490.44a
Non micronutrients foliar application 44.21a 105.47b466.54b839.42b1579.12b1496.71b1411.45b
Fertilizer
Chemical P 47.12b117.74b518.91b899.97b1651.65b1582.93b1433.34b
Biological P × Chemical P
Biological P
No fertilizer
51.43a 142.87a611.38a988.45a1810.41a 1721.75a 1628.78a
43.35c 114.80b467.54c899.28b1641.62b1525.47b1428.32b
35.66d 96.85c 415.46d748.60c1477.86c 1377.93c 1313.33c
1- Columns means followed by the same letter are not significantly different at 0.05 or 0.01 probability level

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29
0
200
400
600
800
1000
1200
1400
1600
1800
2000
35506580 95110125
Day after sowing
Dry matter accumulation (g/m2
Chemical phosphorus
fertilizer
Chemical and biological
phosphorus fertilizers
Biological phosphorus
fertilizer
No fertilizer


Figure 1. Effects of biological and chemical phosphorus fertilizers on dry matter accumulation (g/m2) (Error bars
are the standard error of the mean)



0
200
400
600
800
1000
1200
1400
1600
1800
35 50658095 110125
Day after sowing
Dry matter accumulation (g/m2
Micronitrients
foliar
application
Non foliar
application

Figure 2. Effects of foliar application of micronutrients on dry matter accumulation (g/m2) (Error bars are the
standard error of the mean)