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Effects of organic manure and biological fertilizers on macronutrient uptake of corn (Zea Mays L.).


Corn (Zea mays L.) among the crops, is an important in temperate climatic region, because of the increasing demand for food and livestock feed. The productivity of corn mainly depends on its nutrient management [12]. Macronutrient deficiency in cereals is a reported constraint for plant growth. Nitrogen and phosphorus are essential nutrients for plant growth and development in corn [37]. Nitrogen is one of the most important nutrients for maize production as it affects dry matter production by influencing leaf area development and maintenance as well as photosynthetic efficiency. It can be applied through chemical or organic manure [9,35] and biological means [39,17] but chemical nitrogen fertilizer is expensive. N can be easily lost by leaching, denitrification or volatilization. Agricultural systems require surplus N additions in order to produce desired yields because current management practices tend to disengage energy flows and nutrient cycles in space and time [32]. Phosphorus is second only to nitrogen in mineral nutrients most commonly limiting the growth of terrestrial plants. Ironically, soils may have large reserves of total P, but the amounts available to plants is usually a tiny proportion of this total [8]. However phosphorus after application, a considerable amount of P is rapidly transformed into less available forms (up to 80 %) by forming a complex with Al or Fe in acid soils or Ca in calcareous soils before plant roots have had a chance to absorb it [18]. Large quantities of chemical fertilizes are used to replenish soil N and P, resulting in high costs and severe environmental contamination [7]. As a result, poor farmers have no easy access to chemical fertilizers. Additionally, a high mineral P input supports surface runoff, and P might be lost by soil erosion or leaching [38] which is a waste of the limited P resources and results in eutrophication of rivers, lakes and natural habitats. Environmental protection and the need to enhance agricultural output have made research in new sustainable technologies necessary. Several studies have conclusively shown that PSM solubilizes the fixed soil P and applied phosphates, resulting in higher crop yields [40,26,34,10,29]. The nitrogen fixing bacteria such as Azotobacter and Azosperillium reduce the nitrogen gas to ammonia using intensive energy to break the nitrogen bonds so that it can combine with hydrogen to form ammonia. Many actual and putative are biofertilizer PGPR produce phytohormones that are believed to be related to their ability to stimulate plant growth. Inoculation of plants with this bacterium causes morphological changes, such as an increase in root surface area through the production of more root hairs, which in turn enhance mineral uptake [13,25]. Economic and environmental benefits can include increased income from high yields, reduced fertilizer costs and reduced emission of the greenhouse gas, [N.sub.2]O as well as reduced leaching of N[O.sub.3]. The objective of the present work was to evaluate the efficiency of the interaction between PGPR (Azotobacter and Azosperillium), PSM bacterium (Pseudomonas putida and Bacillus lentus) on the macronutrient uptake by corn.

Materials and Methods

2. 1. Exprimental design:

An experiment was conducted at research farm of Sari Agricultural Sciences and Natural Resources University (Latitude 42. 36N, longitude 13. 53E and 16 m above mean sea level), Iran during 2007. Experiment laid out as split plot based on randomized complete block design with three replications. Three levels of manures (consisted of 20 Mg. [ha.sup.-1] farmyard manure, 15 Mg. [ha.sup.-1] green manure of barley and control or without any manures) as main plots and eight levels of biofertilizers (consisted of 1- NPK or conventional fertilizer application; 2-NPK+PSM+PGPR; 3-[NP.sub.50%]K+PSM+PGPR; 4-[N.sub.50%]PK+PSM+PGPR; 5-[N.sub.50%][P.sub.50%]K+PSM+PGPR; 6-PK+PGPR; 7-NK+PSM and 8-PSM+PGPR) as sub plots were treatments. To chemical analyze, soils and farmyard manure were sampled before the experiments. Results of the soil and farmyard manure analysis are in Table 1 and 2. The fertilizers NPK at concentrations of 300, 120 and 100 kg [ha.sup.-1] were applied in the form of urea, diammonium phosphate and muriate of potash, respectively. All of PK and half of N (starter fertilizer) were mixed with the soil at the time of sowing, while remaining N was applied in solution form at tasseling stage.

2. 2. Bacterial Strain and Inoculation:

The bacterium used in this study were phosphates solubilization microorganisms (Pseudomonas putida, Bacillus lentus) and plant growth promoting rhizohactoria (Azotobacter coroocoocum, Azospirili brasilense). Zea mays cv. SC 604 seeds were surface sterilized with 70% sodium hypochlorite for 1 min at room temperature. Then the seeds were washed repeatedly with sterile distilled water. Bacterial were suspended in suspension of sugar in water. This slurry was used to introduce the bacteria as corn seed coatings.

2. 3. Plant Material:

At harvest in the 2007 growth season, the plants from one 1 [m.sup.2] subplots per field were combined, the grain was removed from the straw, and then the straw and grain weights were recorded. For chemical analysis, plant samples were oven dried at 70[degrees]C for 48 h and ground to a powder (2 mm). Mineral concentrations of above ground tissues (grain and leaf of corn) were determined. Total nitrogen of leaf were determined by Kjeldhal technique (Tecator Kjeltec Auto 1030 analyzer) and phosphorus was determined by using a spectrophotometer (Uvikon 931, Italy) and total potassium by flame photometer (Systronics Mediflame 127).

2. 4. Statistical Analysis:

Data were subjected to ANOVA using the SAS statistical software package using a GLM (SAS Institute, 2000) and Duncan's multiple range test was performed to compare the treatment means. The level of statistical significant was accepted as P<0. 05.

Results and Discussion

Results showed that application of farmyard manure (FYM) increased nitrogen, phosphorus and potassium of grain and phosphorus uptake by leaf corn compared to the control (Table 3). Moreover, green manure (GM) application increased nitrogen and potassium of grain and phosphorus and potassium uptake of leaf compared to the control. NPK came from organic sources, such as FYM and GM can be used as a sole source or as a substitute for part of inorganic fertilizers (Cherr et al., 2006). However, in most situations the total N requirement of the corn crop is supplied by a combination of sources. Soils supply N by organic matter decay, N added to the soil by PGPR and fertilizer.

The data (Table 3) indicated that plant growth promoting rhizohacteria (PGPR) and phosphate solubilization (PSM) inoculation significantly increased nitrogen and potassium of grain and nitrogen, phosphorus and potassium of corn leaf and increased the yield of corn (data not shown) compared to the control. Probably, plant growth promoting rhizobacteria by production of growth stimulating phytohormones [37] mobilization of phosphate [40] siderophore production [39] antibiotic production [11] inhibition of plant ethylene synthesis [33] induction of plant systemic resistances to pathogens [3] and improve growth of root [13] could increase the mineral uptake of plants. According to results in all fertilizer treatments application of PSM and PGPR together could reduce P application by 50% without any significant decrease nitrogen, phosphorus and potassium of grain and nitrogen and phosphorus of corn leaf. The result of this relationship could be due to a positive interaction that may have occurred between PSM and PGPR [40]. Increasing the availability of P and K in soils with inoculation of PSM or with combined inoculation with PSM and PGPR, which may lead to increased P uptake and plant growth, was reported by many researchers [1,40,38]. Bacteria inoculation, which can improve P and K availability in soils by producing organic acids and other chemicals, stimulated growth and mineral uptake of plants [39].

Plant growth promoting rhizobacteria (PGPR) are root colonizing microorganisms which are known to fix atmospheric molecular nitrogen through symbiotic and a symbiotic or associative nitrogen fixing process. The effect of associative microorganisms in increasing crop yield and [N.sub.2] fixation has been reported by many research workers [40,26,34,10]. Lerner et al., [17] reported that inoculation of maize with Azospirillum brasilense resulted in a proliferation of root hairs which could have dramatic effects on increasing root surface area. Indole-3-acetic acid, cytokinins and gibberellins is phytohormones which are known to be involved in root initiation, cell division, and cell enlargement [13]. This hormone is very commonly produced by PGPR [6,10]. The fact that plant growth and nutrient uptake increased in the presence of PGPR and PSM suggested a strong synergistic relationship between root colonization, P uptake and growth promotion. In agreement with these findings, Zaidi et al., [40] observed that in low P soils plant growth and nutrient uptake in chickpea were greater after inoculation with tripartite culture of Mesorhizobium, PSM and G. fasciculatum than after inoculation with each organism alone. In the present study, application of farmyard manure and co-inoculation with PSM and PGPR significantly increased the uptake of phosphorus compared to the control (Figure 1). Nikolay et al., [20] and Orhan et al., [22] reported that the major factors limiting growth of PGPR and PSM in the rhizosphere are the availability of soluble organic substrates. Intuitively, adding organic fertilizers before or simultaneously with inoculation seems an effective method to improve microbial survival [5]. The nutritious substances are introduced together with microorganisms, so that the PGPR inoculants survive at the initial time. In addition, in most cases these phytohormones are believed to be changing assimilate partitioning patterns in plants and affecting growth patterns in roots to result in bigger roots, more branched roots, and/or roots with greater surface area [19].

By contrast reducing of N around 50%, in [N.sub.50][P.sub.50]K+PGPR+PSM treatment, significantly increased phosphorus content in the corn leaf. It probably is affected low levels of available of nitrogen in soils and accumulation of phosphorus in high level. In addition several studies have shown that organic materials and their decomposition products can reduce P fixation in soils. Recently the use of bio-organic fertilizers in agriculture has received considerable attention because of the environmental problems associated with alternative disposal methods and the harmful affect on human health. On the other hands, bio-organic fertilizers improves the physical properties of the soil, increasing water holding capacity, preventing nutrient leaching and adding more mineral nutrients to the poor sandy soil. Application of biological fertilizers in plots with sufficient nutrients and 50% lower P compared to control did not significant effect on leaf phosphorus content.

Using of biological fertilizers and NPK in green manure plots compared to the control plots had higher accumulation of nitrogen in seeds (Figure 2). Such inoculation could compensate for nutrient deficiency and improve a plant development through production of plant growth regulators by microbes at the root interface, which stimulated root development of plants and resulted in better absorption of water and nutrients from the soil [31,5].



Among different parameters, ear leaf nitrogen content at tasseling stage had the most correlation with gain yield. Furthermore, inoculation of PSM in NK+PSM treatment increased of nitrogen in seeds. It is generally thought that PSM in addition to solubilizing inorganic P also release growth-promoting substances [11]. In the present study, a positive correlation between the yield grain and NPK content of corn seed and leaf (Table 4). In the other hand, results demonstrate that there is important relationship between grain yield and the amount of nutrient in corn. Furthermore, nitrogen content have high correlation with potassium content of corn seed (r=0. 81**) that show positive relationship among the nitrogen and potassium content.


The present study clearly indicated that the mixed inoculation of N2 fixing bacterium and PSM improved nutrient uptake. In our case, co-inoculation of PSM and PGPR strains synergistically solubilized P which were added into the soil and make them much more available for uptake by plant root. In addition, the positive effect on plant growth of the bio-inoculants in this study might not only result from a direct PGPR effect but also from an indirect modification of the bacterial community. Moreover, it would be preferable for the PGPR or PSM strains composing the treatments to be adapted to the local management practice. Furthermore, a synergistic effect on the level of organic carbon was observed between PSM and PGPR when they were coinoculated. On the other hand, bio-organic fertilizers improves the physical properties of the soil, increasing water holding capacity, preventing nutrient leaching and adding more mineral nutrients. In addition, these results suggest that plant growth stimulating efficiency of bacterial inoculants affected by soil nutritional condition. In short, results from all these and other experiments suggest that coinoculation of PGPR with different beneficial properties should be the future trends of bio-fertilizer application for sustainable crop production.


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(1) Yazdani Mohammad and (2) Pirdashti. H

(1) Ph.D Student, Department of Agronomy, Tabriz Branch, Islamic Azad University, Tabriz, Iran.

(2) Department of Agriculture, Genetics and Agricultural Biotechnology of Tabarestan, Sari Agricultural Sciences and Natural Resources University.

Corresponding Author

Yazdani Mohammad, Ph.D Student, Department of Agronomy, Tabriz Branch, Islamic Azad University, Tabriz, Iran.

Table 1: soil chemical properties, and soil particle distribution of
the top soil layer (0-30 cm).

                            N              P              K
                    OM      (mg 100        (mg 100        (mg 100
Type         PH     (%)     [gr.sup.-1])   [gr.sup.-1])   [gr.sup.-1])

Silty loam   7. 5   3. 48   193            12. 3          367. 3

             Soil particle size (mm)

Type         2. 0-0. 2   0. 2-0. 02   <0. 02

Silty loam   47. 3       42. 1        10. 6

Table 2: farmyard manure chemical properties.

PH      Ec                Nt       OM
        (dcs/[m.sup.2])   (%)

8. 18   3. 39             2. 038   7. 35

PH      P      K      Cu      Zn       Mn       Fe


8. 18   7. 6   6. 8   3. 78   36. 93   93. 45   45. 39

Table 3: The effect of PGPR and PSM Application on macronutrient
uptake of Corn (Zea mays L.).


Treatments                               (gr [plant.sup.-1])

Organic Manure (A)
Farmyard manure                          1.67a
Green manure                             1.62a
Control                                  1.37b
Biofertilizer (B)
NPK                                      1.67b
NPK+[PGPR.sub.+]PSM                      1.96a
[NP.sub.50]K+[PGPR.sub.+]PSM             1.95a
[N.sub.50]PK+[PGPR.sub.+]PSM             1.43c
[N.sub.50][P.sub.50]K+[PGPR.sub.+]PSM    1.42c
PK+PGPR                                  1.21d
NK+PSM                                   1.64b
PGPR+PSM                                 1.14d
A                                        *
B                                        **
A x B                                    **
CV                                       11.6


Treatments                               (mg [plant.sup.-1])

Organic Manure (A)
Farmyard manure                          22.1a
Green manure                             20.7ab
Control                                  18.6b
Biofertilizer (B)
NPK                                      21.6ab
NPK+[PGPR.sub.+]PSM                      24.3a
[NP.sub.50]K+[PGPR.sub.+]PSM             23.4a
[N.sub.50]PK+[PGPR.sub.+]PSM             21.3ab
[N.sub.50][P.sub.50]K+[PGPR.sub.+]PSM    18.9bc
PK+PGPR                                  17.8c
NK+PSM                                   19.5bc
PGPR+PSM                                 17.0c
A                                        *
B                                        **
A x B                                    NS
CV                                       14.9


Treatments                               (mg [plant.sup.-1])

Organic Manure (A)
Farmyard manure                          41.8a
Green manure                             39.3b
Control                                  36.5c
Biofertilizer (B)
NPK                                      41.6b
NPK+[PGPR.sub.+]PSM                      46.5a
[NP.sub.50]K+[PGPR.sub.+]PSM             46.4a
[N.sub.50]PK+[PGPR.sub.+]PSM             36.6cd
[N.sub.50][P.sub.50]K+[PGPR.sub.+]PSM    37.3bcd
PK+PGPR                                  34.2de
NK+PSM                                   39.7bc
PGPR+PSM                                 31.1e
A                                        **
B                                        **
A x B                                    NS
CV                                       11.5

                                         1 Leaf

Treatments                               (gr [plant.sup.-1])

Organic Manure (A)
Farmyard manure                          0.53a
Green manure                             0.51a
Control                                  0.50a
Biofertilizer (B)
NPK                                      0.57bc
NPK+[PGPR.sub.+]PSM                      0.66a
[NP.sub.50]K+[PGPR.sub.+]PSM             0.63ab
[N.sub.50]PK+[PGPR.sub.+]PSM             0.46de
[N.sub.50][P.sub.50]K+[PGPR.sub.+]PSM    0.49cd
PK+PGPR                                  0.41ef
NK+PSM                                   0.54cd
PGPR+PSM                                 0.37f
A                                        NS
B                                        **
A x B                                    NS
CV                                       14.8

                                         1 Leaf

Treatments                               (mg [plant.sup.-1])

Organic Manure (A)
Farmyard manure                          127.4a
Green manure                             107.0b
Control                                  83.1c
Biofertilizer (B)
NPK                                      105.2c
NPK+[PGPR.sub.+]PSM                      140.6a
[NP.sub.50]K+[PGPR.sub.+]PSM             124.5ab
[N.sub.50]PK+[PGPR.sub.+]PSM             104.0c
[N.sub.50][P.sub.50]K+[PGPR.sub.+]PSM    109.4bc
PK+PGPR                                  93.3cd
NK+PSM                                   101.7cd
PGPR+PSM                                 85.0d
A                                        **
B                                        **
A x B                                    *
CV                                       15.2

                                         1 Leaf

Treatments                               (gr [plant.sup.-1])

Organic Manure (A)
Farmyard manure                          2.15ab
Green manure                             2.31a
Control                                  1.82b
Biofertilizer (B)
NPK                                      2.07bc
NPK+[PGPR.sub.+]PSM                      2.66a
[NP.sub.50]K+[PGPR.sub.+]PSM             2.15bc
[N.sub.50]PK+[PGPR.sub.+]PSM             1.74c
[N.sub.50][P.sub.50]K+[PGPR.sub.+]PSM    2.21b
PK+PGPR                                  2.12bc
NK+PSM                                   2.06bc
PGPR+PSM                                 1.96bc
A                                        NS
B                                        *
A x B                                    NS
CV                                       18.4

Levels of significant: * P< %5, ** P<%1, NS = not significant

Table 4: Correlation matrix for yield and N, P, K content of seed and

    Yield     Seed                          Leaf

              N         P         K         N         P         K

1   1         2         3         4         5         6         7
2   0.81 **   1
3   0.80 **   0.61 **   1
4   0.99 **   0.81 **   0.80 **   1
5   0.80 **   0.76 **   0.66 **   0.80 **   1
6   0.63 **   0.65 **   0.65 **   0.63 **   0.58 **   1
7   0.38 **   0.42 **   0.45 **   0.38 **   0.44 **   0.34 **   1

Levels of significant: * P< %5, ** P<%1, NS = not significant
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Title Annotation:Original Article
Author:Mohammad, Yazdani; Pirdashti, H.
Publication:Advances in Environmental Biology
Article Type:Report
Geographic Code:7IRAN
Date:Dec 1, 2011
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