Advancement of Nitrogen Fertilization on Tropical Environmental

*Elizeu Monteiro Pereira Junior, Elaine Maria Silva Guedes Lobato, Beatriz Martineli Lima, Barbara Rodrigues Quadros, Allan Klynger da Silva Lobato, Izabelle Pereira Andrade and Letícia de Abreu Faria*

#### **Abstract**

The nitrogen (N) fertilization synthetic or biological is primordial for food production worldwide. The consumption of N fertilizers in agricultural systems increased in exponential scale, mainly in developing countries. However, some negative points are associated to industrial N consumption; consequently the industry promoted ways to minimize N losses in production systems of tropical agriculture. Biological nitrogen fixation is a very important natural and sustainable process for the growth of leguminous plants, in which many micronutrients are involved, mainly as enzyme activators or prosthetic group. However, other mechanisms in the rhizosphere and molecular region still need to be clarified. Therefore, the aim of this chapter is to compile information about the historical and current affairs about the advances in N fertilization in tropical environments through a history from N fertilization world-wide, N balance in the main agricultural systems, introduction of alternative ways to avoid N losses, advances between BNF and micronutrients, as well as the effects of N absence in plant metabolisms. Biological nitrogen fixation is a very important natural process for the growth of leguminous plants, in addition many metallic nutrients, micronutrients, are involved in BNF metabolism, mainly as enzyme activators or prosthetic group. But other mechanisms in the rhizosphere and molecular region still need to be clarified.

**Keywords:** ammonia synthesis, biological N fixation, humic substances, N balance, volatilization

#### **1. Introduction**

Hellriegel and Wilfarth showed definitive evidence for N2 fixation by microbes in legumes in 1886, but the industrial process to fertilizer production known as the Haber-Bosch was established just in 1906, which uses a catalytic agent at high pressure and high temperature [1].

Actually, the world population has now been increasingly relying on nitrogen (N) fertilizers in order to keep up with the demands of food and economic growth rates; on the other hand, less than 30% of synthetic fertilizers would actually be

utilized; the unused chemicals sprayed on crops would be lost in the field and could subsequently cause serious environmental problems.

Urea is a popular N source in developing countries due to its advantages of a high N content, safety, and easy transportation [2]. However, the increase of pH and surface soil NH4 + concentrations resulting from urea hydrolysis can exacerbate NH3 emission.

This causes low N use efficiency, especially in alkaline soils or soils with low sorption capacity, which limits the use of urea fertilizer in Europe [3]. In tropical areas, increasing the adoption of no-tillage systems also induces to high N losses from urea fertilization, in tropical soils, due to high temperatures and moisture; NH3 losses exceeding 40% of the surface-applied urea N have been reported, especially under no-till or perennial crops where plant residues are kept on the soil surface [4].

Nitrogen losses by NH3 emission not only brings economic loss to farmers, but also detrimental effects to ecosystems and human health, while the biological nitrogen fixation (BNF) has the advantage of being environmentally friendly and therefore would be ideal for sustainable agriculture.

Enormous progress in almost all aspects of BNF has been made in the past century, especially in the recent two decades, in genetics and biochemistry, culminating in the determination of the crystallographic structures of both nitrogenase components and micronutrients metabolism.

These information collaborated to elucidate N assimilation routes in plants clarifying further its essentiality and allowing to infer that plants can be affected negatively in molecular even genetic level in N absence.

Therefore, the aim of this chapter is to compile information about the historical and current concerns about the advances in N fertilization in tropical environments through a history from N fertilization worldwide, N balance in the main agricultural systems, introduction of alternatives ways to avoid N losses, advances between BNF and micronutrients, as well as the effects of N absence in plant metabolism.

#### **2. History of nitrogen fertilization on tropical environmental**

Nitrogen is an essential element to all organisms, because it is part of protein, acids, and other organic compounds [5]. The importance of this nutrient for plants is already known since the 1660s; however, only at 1804 De Saussure received credits for N essentiality after observations of nitrate uptake from soil solution. In this same period, other researchers, as Liebig at 1840, fortified the idea of plants absorb N from atmosphere [6, 7].

Around 78% of the atmosphere gas is compound for N however in gaseous form chemically unavailable. In front of the increased demand by food production and need of N restitution after crop harvests, Fritz Haber at 1909 synthetizes the gaseous element to ammonia (NH3) through a reaction with hydrogen and iron on high pressure and temperatures, which posteriorly was industrially developed by Carl Bosch in 1912–1913, resulting at the known Haber-Bosch process [8].

The N sources used on agricultural activities, even at the end of the eighteenth century, were from crop residues and animal manure modified or not through composting. The production and management of N fertilizers to increase crop yield, as well as corn [9, 10] and wheat [11] around the world [12] have begun at the Green Revolution of the nineteenth century, followed by ammonia synthesis in the beginning of the twentieth century and the increased need of high yield on agricultural areas [13].

**95**

**Table 2.**

*Advancement of Nitrogen Fertilization on Tropical Environmental*

**World 2015 2016 2017 2018 2019 2020 Reference** Total capacity NH3 174.781 181.228 185.222 186.804 186.920 188.310 [11]

Total capacity NH3 8.310 9.545 10.739 10.700 10.700 11.000

Total capacity NH3 24.301 27.618 28.688 29.304 29.320 29.346

Total capacity NH3 99.959 101.188 101.703 101.734 101.734 102.799

Total capacity NH3 40.378 41.044 42.338 43.211 43.311 43.311

Total capacity NH3 1.833 1.833 1.854 1.854 1.854 1.854

Animal feces 56% 40% 40% Synthetic fertilizers 22% 44% 46%

Animal manure applied in soil 4% 4% 4% Animal manure left in pasture 91% 84% 84% Synthetic fertilizers 5% 12% 12%

Animal manure applied in soil 16% 13% 13% Animal manure left in pasture 60% 50% 47% Synthetic fertilizers 24% 37% 40%

Animal manure applied in soil 20% 13% 12% Animal manure left in pasture 61% 34% 30% Synthetic fertilizers 19% 53% 58%

Animal manure applied in soil 40% 28% 30% Animal manure left in pasture 27% 17% 17% Synthetic fertilizers 33% 55% 53%

Animal manure applied in soil 2% 3% 4% Animal manure left in pasture 96% 91% 77% Synthetic fertilizers 2% 6% 19% *The percentual represents averages from the 1960s, 1980s, and 2000s (adapted of FAO [12]).*

*Global cumulative of N fertilization from animal manure and fertilizers between 1961 and 2014.*

*Estimative of supply capacity of N (NH3) in continents (in thousand tons) of 2015–2020 (adapted of FAO [12]).*

**World 1960 1980 2000 Reference** Animal manure applied in soil 22% 16% 14% [14]

*DOI: http://dx.doi.org/10.5772/intechopen.90699*

**Africa**

**Americas**

**Asia**

**Europe**

**Oceania**

**Table 1.**

**Africa**

**Americas**

**Asia**

**Europe**

**Oceania**

*Advancement of Nitrogen Fertilization on Tropical Environmental DOI: http://dx.doi.org/10.5772/intechopen.90699*


**Table 1.**

*Nitrogen Fixation*

surface soil NH4

emission.

surface [4].

metabolism.

absorb N from atmosphere [6, 7].

+

utilized; the unused chemicals sprayed on crops would be lost in the field and could

N content, safety, and easy transportation [2]. However, the increase of pH and

This causes low N use efficiency, especially in alkaline soils or soils with low sorption capacity, which limits the use of urea fertilizer in Europe [3]. In tropical areas, increasing the adoption of no-tillage systems also induces to high N losses from urea fertilization, in tropical soils, due to high temperatures and moisture; NH3 losses exceeding 40% of the surface-applied urea N have been reported, especially under no-till or perennial crops where plant residues are kept on the soil

Nitrogen losses by NH3 emission not only brings economic loss to farmers, but also detrimental effects to ecosystems and human health, while the biological nitrogen fixation (BNF) has the advantage of being environmentally friendly and

Enormous progress in almost all aspects of BNF has been made in the past century, especially in the recent two decades, in genetics and biochemistry, culminating in the determination of the crystallographic structures of both nitrogenase

These information collaborated to elucidate N assimilation routes in plants clarifying further its essentiality and allowing to infer that plants can be affected

and current concerns about the advances in N fertilization in tropical environments through a history from N fertilization worldwide, N balance in the main agricultural systems, introduction of alternatives ways to avoid N losses, advances between BNF and micronutrients, as well as the effects of N absence in plant

**2. History of nitrogen fertilization on tropical environmental**

Therefore, the aim of this chapter is to compile information about the historical

Nitrogen is an essential element to all organisms, because it is part of protein, acids, and other organic compounds [5]. The importance of this nutrient for plants is already known since the 1660s; however, only at 1804 De Saussure received credits for N essentiality after observations of nitrate uptake from soil solution. In this same period, other researchers, as Liebig at 1840, fortified the idea of plants

Around 78% of the atmosphere gas is compound for N however in gaseous form chemically unavailable. In front of the increased demand by food production and need of N restitution after crop harvests, Fritz Haber at 1909 synthetizes the gaseous element to ammonia (NH3) through a reaction with hydrogen and iron on high pressure and temperatures, which posteriorly was industrially developed by

The N sources used on agricultural activities, even at the end of the eighteenth century, were from crop residues and animal manure modified or not through composting. The production and management of N fertilizers to increase crop yield, as well as corn [9, 10] and wheat [11] around the world [12] have begun at the Green Revolution of the nineteenth century, followed by ammonia synthesis in the beginning of the twentieth century and the increased need of high yield on agricultural

Carl Bosch in 1912–1913, resulting at the known Haber-Bosch process [8].

Urea is a popular N source in developing countries due to its advantages of a high

concentrations resulting from urea hydrolysis can exacerbate NH3

subsequently cause serious environmental problems.

therefore would be ideal for sustainable agriculture.

negatively in molecular even genetic level in N absence.

components and micronutrients metabolism.

**94**

areas [13].

*Estimative of supply capacity of N (NH3) in continents (in thousand tons) of 2015–2020 (adapted of FAO [12]).*


#### **Table 2.**

*Global cumulative of N fertilization from animal manure and fertilizers between 1961 and 2014.*

Data from the FAO [14] estimated that the global capacity of N ammonia offer increases annually of 1.5% in average, with production of 174,781–188,310 thousands of tons of 2015–2020 (**Table 1**).

In addition, during this period, Africa, Oceania, Europe, and the Americas increased the capacity to 32.4, 1.1, 7.3, and 20.8%, respectively, however, stands out to Asia continent with the highest productive capacity estimated to 102,799 thousands of tons of N to 2020 (**Table 1**).

Estimates in global scale from FAOSTAT [15] show N inputs from animal manure increased from 66 to 113 million from 1961 to 2014, while N fertilizers applied in soils increased from 18 to 28 million of tons of N, respectively.

The use of N fertilizer at Europe continent increased 33% (about 5 million of tons of N), as a similar tendency observed in others regions (**Table 2**).

Brazil is one of the biggest fertilizer consumers in the world. The significant increase in fertilizer consumption occurred between 1988 and 2010 [16] as consequence of public policy implementation and Brazilian agriculture modernization.

Nitrogen had a higher growth consumption among the nutrients from NPK in the analyzed period, around 250%, from 814,952 to 2,854,189 tons; however, N fertilizers consumption was 12,211,855 ton from 2010 to 2013 and to around 15,469,549 tons from 2014 to 2017 [17, 18].

#### **3. Nitrogen balance in the tropical agricultural systems**

Nitrogen balance in the systems becomes a concern for tropical agriculture as a result of the high scale of N fertilizer production. Nutrient balance is a parameter that analyzes the relation between quantity of vegetable biomass produced and nutrient applied. Besides, nutrient balance is a tool with easy application and able to guide the management to efficient fertilization [19].

Nitrogen balance as a management technique accounts the nutrient exportation by crops, residual in soil and the N losses [20]; thus it is essential to a balanced fertilization strategy aiming to maximize the economic return and ensure the environmental quality.

The calculations of nutrient balance evaluation must account for the input and output of N because this nutrient can be distributed by soil, plant, and animal (**Table 3**). Between 95 and 100% of the total N input into soil is from the surface through rainfall or dust and aerosols, irrigation, runoff and groundwater, biological fixation by phototrophic and heterotrophic organisms, organic and inorganic fertilization, and seed reserves. Besides the plants exports, the N output occurs by erosion, leaching and drainage, ammonia volatilization, denitrification, and senescence plants [21, 22].

Brazilian crop exports 50% of N in harvested product mainly by the largest exportations of soybean (70%), corn (15%), sugarcane (8%), rice (2%), and wheat (2%) [17]. However, these N quantities have contribution from the N biologic fixation (NBF), mainly from soybean with 82% of the total N input in crops production.

Soybean occupied the largest area of agriculture in Brazil between 2013 and 2016 and also was responsible for the largest nutrient exportation, although N is not applied in this crop, it comprised 70% of the total N exported by all crops, while phosphorus and potassium reached 57.5 and 56.8%, respectively [23]. Analyze nutrient exportation nutrient exportation for area unity in this period was found out the largest nutrient exporters were soybean (181 kg ha<sup>−</sup><sup>1</sup> ), tomato (159 kg ha<sup>−</sup><sup>1</sup> ), and cotton (129 kg ha<sup>−</sup><sup>1</sup> ).

**97**

ture, and humidity [29–34].

*Advancement of Nitrogen Fertilization on Tropical Environmental*

**Total N input** X = A + B + C + D + E + F + G

**Total N output** Y=I + J + K + L + M + N + O + P

M

**Source Amount References N input** [3]

**4. Ways to avoid N losses from agricultural systems**

atmosphere by denitrification and ammonia volatilization [24, 25].

In agricultural systems there are losses in general; however, N losses are considered highly relevant [24, 25]. Nitrogen losses are a potential contaminant and can impact production cost. Nitrogen is a dynamic element in soil and can be lost to the

*N balance from total of inputs and outputs and N in soil in the beginning and final of agricultural* 

Nb/Years of experiment

Ammonia volatilization is a concerning problem because it represents high N losses in soil–plant system besides to be a threat for global environmental [26], while the N losses by denitrification in tropical areas are less significant in consequence of its restriction in the use of nitrate as fertilizer due its explosive potential [25–28]. Global agricultural production is responsible for 50% of N losses by ammonia volatilization meaning 37 tons of N for year; however, the losses can be higher according to the N source, application way, soil management, climate, soil tempera-

*DOI: http://dx.doi.org/10.5772/intechopen.90699*

Total N fertilization rates A Total manure applied B N symbiotic fixation C Atmospheric deposition of N D Irrigation water E N input by seed in harvest F N nonsymbiotic fixation G

N exportation in crops and/or biomass I N losses by denitrification J N losses by ammonia volatilization K N losses by plants senescence L

N losses by surface runoff N N leaching O N losses by soil erosion P

Total N in beginning of the experiment Q Total N in end of the experiment R **Total N changes in soil** Z = R-Q

During experiment performance Nb = X-Y-Z

Gaseous losses of N (except NH3

**N output**

volatilization)

**Total N in soil**

**N balance**

**Table 3.**

**N balance for year (kg N ha−1 yr−1)**

*experiments, modified from [3].*


#### *Advancement of Nitrogen Fertilization on Tropical Environmental DOI: http://dx.doi.org/10.5772/intechopen.90699*

#### **Table 3.**

*Nitrogen Fixation*

modernization.

tons from 2014 to 2017 [17, 18].

environmental quality.

senescence plants [21, 22].

and cotton (129 kg ha<sup>−</sup><sup>1</sup>

sands of tons of 2015–2020 (**Table 1**).

sands of tons of N to 2020 (**Table 1**).

Data from the FAO [14] estimated that the global capacity of N ammonia offer increases annually of 1.5% in average, with production of 174,781–188,310 thou-

In addition, during this period, Africa, Oceania, Europe, and the Americas increased the capacity to 32.4, 1.1, 7.3, and 20.8%, respectively, however, stands out to Asia continent with the highest productive capacity estimated to 102,799 thou-

Estimates in global scale from FAOSTAT [15] show N inputs from animal manure increased from 66 to 113 million from 1961 to 2014, while N fertilizers applied in soils increased from 18 to 28 million of tons of N, respectively.

tons of N), as a similar tendency observed in others regions (**Table 2**).

**3. Nitrogen balance in the tropical agricultural systems**

guide the management to efficient fertilization [19].

The use of N fertilizer at Europe continent increased 33% (about 5 million of

Nitrogen had a higher growth consumption among the nutrients from NPK in the analyzed period, around 250%, from 814,952 to 2,854,189 tons; however, N fertilizers consumption was 12,211,855 ton from 2010 to 2013 and to around 15,469,549

Nitrogen balance in the systems becomes a concern for tropical agriculture as a result of the high scale of N fertilizer production. Nutrient balance is a parameter that analyzes the relation between quantity of vegetable biomass produced and nutrient applied. Besides, nutrient balance is a tool with easy application and able to

Nitrogen balance as a management technique accounts the nutrient exportation

The calculations of nutrient balance evaluation must account for the input and output of N because this nutrient can be distributed by soil, plant, and animal (**Table 3**). Between 95 and 100% of the total N input into soil is from the surface through rainfall or dust and aerosols, irrigation, runoff and groundwater, biological fixation by phototrophic and heterotrophic organisms, organic and inorganic fertilization, and seed reserves. Besides the plants exports, the N output occurs by erosion, leaching and drainage, ammonia volatilization, denitrification, and

Brazilian crop exports 50% of N in harvested product mainly by the largest exportations of soybean (70%), corn (15%), sugarcane (8%), rice (2%), and wheat (2%) [17]. However, these N quantities have contribution from the N biologic fixation (NBF), mainly from soybean with 82% of the total N input in crops

Soybean occupied the largest area of agriculture in Brazil between 2013 and 2016 and also was responsible for the largest nutrient exportation, although N is not applied in this crop, it comprised 70% of the total N exported by all crops, while phosphorus and potassium reached 57.5 and 56.8%, respectively [23]. Analyze nutrient exportation nutrient exportation for area unity in this period was found

), tomato (159 kg ha<sup>−</sup><sup>1</sup>

),

out the largest nutrient exporters were soybean (181 kg ha<sup>−</sup><sup>1</sup>

).

by crops, residual in soil and the N losses [20]; thus it is essential to a balanced fertilization strategy aiming to maximize the economic return and ensure the

Brazil is one of the biggest fertilizer consumers in the world. The significant increase in fertilizer consumption occurred between 1988 and 2010 [16] as consequence of public policy implementation and Brazilian agriculture

**96**

production.

*N balance from total of inputs and outputs and N in soil in the beginning and final of agricultural experiments, modified from [3].*

#### **4. Ways to avoid N losses from agricultural systems**

In agricultural systems there are losses in general; however, N losses are considered highly relevant [24, 25]. Nitrogen losses are a potential contaminant and can impact production cost. Nitrogen is a dynamic element in soil and can be lost to the atmosphere by denitrification and ammonia volatilization [24, 25].

Ammonia volatilization is a concerning problem because it represents high N losses in soil–plant system besides to be a threat for global environmental [26], while the N losses by denitrification in tropical areas are less significant in consequence of its restriction in the use of nitrate as fertilizer due its explosive potential [25–28].

Global agricultural production is responsible for 50% of N losses by ammonia volatilization meaning 37 tons of N for year; however, the losses can be higher according to the N source, application way, soil management, climate, soil temperature, and humidity [29–34].


**Table 4.**

*Examples of ammonia volatilization due to urea application in different soils, modified from [20].*

Urea is the most N source used in the world; however, also it has high susceptibility to be lost in agricultural systems [24, 25]. The high presence of urease enzyme in soil causes a rapid hydrolysis of urea and, consequently, ammonia losses to the atmosphere [35].

Variable quantities of ammonia lost to the atmosphere were related by urea use in agriculture [35–37] according to the exemplified in **Table 4**.

Urease is an extracellular enzyme naturally presents in soil, plants, and microorganisms acting as a catalyzer of urea in the hydrolysis process [30–32]. This chemical process induces excess of protons (H+ ); consequently it rises pH in soil around the fertilizer granules of 6.5–8.8 or until 9.0 causing unbalance between ammonium (NH4 + ) and ammonia (NH3) [33, 34].

During hydrolysis ammonium carbonate is formed, which is dissociated to produce ammonia ions and hydroxide; however, the relative concentration of ammonia and ammonium is determined by the pH in soil solution, and ammonia is favored under high pH condition according to equations [28].

$$\text{NH}\_4^+ \text{ 4 OH}^- \leftrightarrow \text{NH}\_3 + \text{H}\_2\text{O} \tag{1}$$

**99**

*Advancement of Nitrogen Fertilization on Tropical Environmental*

40 phosphorotriamides synthetized considered the most effective compound to urease inhibition because its composition comprises a functional group containing P=O or P=S bonded for at least one free amide (NH2) to react with urease active

Urease inhibitor known as NBPt (N-(n-butyl) thiophosphoric triamide) has been the most used additives in Brazil, in which urea is the most used N source.

This additive is dissolved in a nonaqueous solvent to adding characteristics as (i) larger stability to NBPt molecule under temperature, humidity and transportation variances, and (ii) higher solubility; (iii) improves adherence of mix solvent + NBPt to urea granule, (iv) low toxicity, and inflammable potential; and (v) acts as buffer agent to keep alkaline pH similar to hydrolysis environment of urea in soil providing

The largest of compounds used along with urea are low efficient when applied in soil [43]. NBPt aim is to retard the ammonia volatilization peak [46]. Generally, chemical compounds with similar structure as urea can be more efficient to retard the volatilization; thus, the bond sites and length of amide of phosphoryl triaside

Recently, lab researches reported beneficial and/or synergic effects of the humic substances use with urea [47–49]; however, the action mechanism is still unknown [49]; also depending of humic substances, the results can be contradictory [50, 51], but there are hypotheses that urease enzymes reduce with the association of humic acid and urea [48]; besides it minimizes N losses, it can improve

Urease inhibitor and humic substances with urea at adjusted pH (pH = 7) provided reduction of 50% from total N volatilization on a Latossolo Vermelho on

**5. Interaction between biological N fixation (BNF) and micronutrients** 

Biological N fixation (BNF) is an important process to global agricultural systems. This phenomenon was discovered in the mid of the nineteenth century by the German chemist Hermann Hellriegel (1831–1895); however, factors on root nodules were unknown, until the Dutch microbiologist and botanic Martinus Beijerinck (1851–1931) identifies microorganisms on root nodules able to realize chemical process to transform atmospherically N to ammonia allowing fixation and absorp-

Fixation biological of N2 (BNF) through the bacteria from genus *Bradyrhizobium* can supply N quantity necessary in legume crops as soybean, besides it is currently observed for many researchers as a clean technology contributing to replace mineral

Nitrogen fixation by bacteria already is well described [56]; however, currently studies are focused in nutrients involved in this metabolism, especially micronutrients [57, 58]. Among the micronutrients able to influence the BNF are boron, copper, zinc, cobalt, iron, nickel, manganese, and molybdenum, essential as structural

Iron is necessary to the production of cofactor FeMo that acts along with nitrogenase enzymes, which can affect significantly the BNF [60]. Excess or default of zinc and nickel can affect the established bacteria inside of the nodules and its

There was an increase in BNF and N uptake as a result of the growth of nodules in number and mass with boron foliar application, and these results were attributed

tion by plants, proving the symbiosis between legumes and bacterial [54].

are similar to urea; however, there are no substrates for urease [45].

*DOI: http://dx.doi.org/10.5772/intechopen.90699*

sites and they are considered [45].

NBPt stability [43].

buffer effect in soil pH [52].

sugar cane [53].

**to higher plants**

N fertilizers in legume crops [55].

symbiosis with plants [57].

components and enzyme activators in plants [56–59].

$$\text{(NH}\_2\text{)}\text{ 2CO} + 2\text{H}\_2\text{O} \rightarrow \text{(NH}\_4\text{)}\text{ 2CO}\_3 \rightarrow \text{NH}\_4\text{ + NH}\_3\uparrow + \text{CO}\_2\uparrow\\\text{OH}^-\tag{2}$$

Researches about urease inhibition in soil have begun over than 70 years ago, resulting in many compounds evaluated and patented as urease inhibitors [38]. Urease has a great effect on the soil-plant system through plant N efficiency, as well as being a versatile enzyme, presenting technological, biotechnological and transgenic applications [39].

Nitrogen losses can be avoided or reduced through organic or inorganic chemical compounds included in urea as an able technology to increase the efficiency of N fertilization at low cost [40–42]. Urea with urease inhibitor can cost around 30% higher than conventional urea [43].

The phosphorotriamides, hydroquinone, catechol, copper, boron, and zinc are the most evaluated additives as urease inhibitor [44]. There are more than

*Advancement of Nitrogen Fertilization on Tropical Environmental DOI: http://dx.doi.org/10.5772/intechopen.90699*

*Nitrogen Fixation*

Grassland soils

Arable soils 50

atmosphere [35].

genic applications [39].

higher than conventional urea [43].

(NH4 +

**Table 4.**

Urea is the most N source used in the world; however, also it has high susceptibility to be lost in agricultural systems [24, 25]. The high presence of urease enzyme in soil causes a rapid hydrolysis of urea and, consequently, ammonia losses to the

*Examples of ammonia volatilization due to urea application in different soils, modified from [20].*

**Mean % N volatilized**

15–200 17.6 New Zealand 50 36.0 USA 30–150 26.7 UK 25 7.5 New Zealand

> 55 30

 77 Brazil 17.8 Denmark 200 30 India 7.9 Argentina 23 Australia

180 22.8 Argentina [16–22]

**Location Reference**

[11, 17, 23–29]

Variable quantities of ammonia lost to the atmosphere were related by urea use

Urease is an extracellular enzyme naturally presents in soil, plants, and microorganisms acting as a catalyzer of urea in the hydrolysis process [30–32]. This chemi-

the fertilizer granules of 6.5–8.8 or until 9.0 causing unbalance between ammonium

During hydrolysis ammonium carbonate is formed, which is dissociated to produce ammonia ions and hydroxide; however, the relative concentration of ammonia and ammonium is determined by the pH in soil solution, and ammonia is favored

Researches about urease inhibition in soil have begun over than 70 years ago, resulting in many compounds evaluated and patented as urease inhibitors [38]. Urease has a great effect on the soil-plant system through plant N efficiency, as well as being a versatile enzyme, presenting technological, biotechnological and trans-

Nitrogen losses can be avoided or reduced through organic or inorganic chemical compounds included in urea as an able technology to increase the efficiency of N fertilization at low cost [40–42]. Urea with urease inhibitor can cost around 30%

The phosphorotriamides, hydroquinone, catechol, copper, boron, and zinc are the most evaluated additives as urease inhibitor [44]. There are more than

); consequently it rises pH in soil around

Brazil Brazil

<sup>+</sup> + OH<sup>−</sup> <sup>↔</sup> NH3 + H2O (1)

<sup>+</sup> + NH3↑ + CO2 + OH<sup>−</sup> (2)

in agriculture [35–37] according to the exemplified in **Table 4**.

**Rate of applied (kg N ha<sup>−</sup><sup>1</sup> )**

150

cal process induces excess of protons (H+

) and ammonia (NH3) [33, 34].

under high pH condition according to equations [28].

(NH2) 2CO + 2H2O → (NH4) 2CO3 → NH4

NH4

**98**

40 phosphorotriamides synthetized considered the most effective compound to urease inhibition because its composition comprises a functional group containing P=O or P=S bonded for at least one free amide (NH2) to react with urease active sites and they are considered [45].

Urease inhibitor known as NBPt (N-(n-butyl) thiophosphoric triamide) has been the most used additives in Brazil, in which urea is the most used N source.

This additive is dissolved in a nonaqueous solvent to adding characteristics as (i) larger stability to NBPt molecule under temperature, humidity and transportation variances, and (ii) higher solubility; (iii) improves adherence of mix solvent + NBPt to urea granule, (iv) low toxicity, and inflammable potential; and (v) acts as buffer agent to keep alkaline pH similar to hydrolysis environment of urea in soil providing NBPt stability [43].

The largest of compounds used along with urea are low efficient when applied in soil [43]. NBPt aim is to retard the ammonia volatilization peak [46]. Generally, chemical compounds with similar structure as urea can be more efficient to retard the volatilization; thus, the bond sites and length of amide of phosphoryl triaside are similar to urea; however, there are no substrates for urease [45].

Recently, lab researches reported beneficial and/or synergic effects of the humic substances use with urea [47–49]; however, the action mechanism is still unknown [49]; also depending of humic substances, the results can be contradictory [50, 51], but there are hypotheses that urease enzymes reduce with the association of humic acid and urea [48]; besides it minimizes N losses, it can improve buffer effect in soil pH [52].

Urease inhibitor and humic substances with urea at adjusted pH (pH = 7) provided reduction of 50% from total N volatilization on a Latossolo Vermelho on sugar cane [53].

#### **5. Interaction between biological N fixation (BNF) and micronutrients to higher plants**

Biological N fixation (BNF) is an important process to global agricultural systems. This phenomenon was discovered in the mid of the nineteenth century by the German chemist Hermann Hellriegel (1831–1895); however, factors on root nodules were unknown, until the Dutch microbiologist and botanic Martinus Beijerinck (1851–1931) identifies microorganisms on root nodules able to realize chemical process to transform atmospherically N to ammonia allowing fixation and absorption by plants, proving the symbiosis between legumes and bacterial [54].

Fixation biological of N2 (BNF) through the bacteria from genus *Bradyrhizobium* can supply N quantity necessary in legume crops as soybean, besides it is currently observed for many researchers as a clean technology contributing to replace mineral N fertilizers in legume crops [55].

Nitrogen fixation by bacteria already is well described [56]; however, currently studies are focused in nutrients involved in this metabolism, especially micronutrients [57, 58]. Among the micronutrients able to influence the BNF are boron, copper, zinc, cobalt, iron, nickel, manganese, and molybdenum, essential as structural components and enzyme activators in plants [56–59].

Iron is necessary to the production of cofactor FeMo that acts along with nitrogenase enzymes, which can affect significantly the BNF [60]. Excess or default of zinc and nickel can affect the established bacteria inside of the nodules and its symbiosis with plants [57].

There was an increase in BNF and N uptake as a result of the growth of nodules in number and mass with boron foliar application, and these results were attributed to the role of boron in the induction of nitrate assimilation by increasing protein synthesis by plant [58].

Manganese has direct role on many enzymatic processes on the BNF, including amide hydrolase enzyme which is directly dependent of Mn+2, and it is responsible for ureide degradation being able to control the BNF under hydric deficiency [61].

Low copper affects the nodule formation and reduces the quantity of fixation bacteria; this element is essential for both bacteria and plants; however, its direct role on BNF is still unclear [59].

Molybdenum is an essential nutrient to BNF taking part on nitrate reductase with the reduction of nitrate (NO3 <sup>−</sup>) to nitrite (NO2 <sup>−</sup>) and on the nitrogenase process in conversion of dinitrogen (N2) to ammonia (NH3) by fixation bacteria. The low quantity demand of molybdenum allows its application on soil and foliar or even by seeds treatments, which is a form of quality aggregation to the seeds by affecting positively on germination [62].

Cobalt is a component of cobalamin and leghemoglobin synthesis, which is controlling their levels on nodules and avoiding nitrogenase enzyme inactivation; thus this element can be considered essential to N2 fixation [63].

Nickel can affect directly the presence and quantity of fixation microorganisms because it is a hydrogenase component (Ni-Fe), which can recycle H2 that is

#### **Figure 1.**

*Root nodules from legume. A1, longitudinal section; A2, approximated image on nodules developed with no Ni; B1,longitudinal section; B2, approximated image on nodules developed with 0.5 g dm<sup>−</sup><sup>3</sup> of Ni; C1, longitudinal section; C2, approximated image on nodules developed with 10 g dm<sup>−</sup><sup>3</sup> of Ni [64].*

**101**

**Figure 2.**

*to nitrogen deficiency [65].*

*Advancement of Nitrogen Fertilization on Tropical Environmental*

generated from N reduction and could affect positively or negatively the legume metabolism [64]. Nickel balance on BNF can be seen on fixation nodules where in

Even though the essentiality had been established for N at higher plants, there are still remained doubts about how the N absence can affect the metabolism. Recently, by modern techniques and sensible equipment, it was possible to deter-

The N deficiency exposure of *Olea europaea* plants was described as a significant decrease on chlorophyll a and net photosynthetic rate (**Figure 2**). Photosynthesis is a process that involves light absorption by the photosynthetic pigments present in

*Chlorophyll a (Chl a), nitrogen content, and net photosynthetic rate (Amax) in* Olea europaea *plants exposed* 

its absence causes large formation of internal cells according to **Figure 1**.

**6. Recent reports about N absence on plants metabolism**

mine clearly as N absence affects plant metabolism and production.

*DOI: http://dx.doi.org/10.5772/intechopen.90699*

*Nitrogen Fixation*

synthesis by plant [58].

role on BNF is still unclear [59].

with the reduction of nitrate (NO3

affecting positively on germination [62].

**100**

**Figure 1.**

*Root nodules from legume. A1, longitudinal section; A2, approximated image on nodules developed with no Ni;* 

to the role of boron in the induction of nitrate assimilation by increasing protein

Manganese has direct role on many enzymatic processes on the BNF, including amide hydrolase enzyme which is directly dependent of Mn+2, and it is responsible for ureide degradation being able to control the BNF under hydric deficiency [61]. Low copper affects the nodule formation and reduces the quantity of fixation bacteria; this element is essential for both bacteria and plants; however, its direct

Molybdenum is an essential nutrient to BNF taking part on nitrate reductase

process in conversion of dinitrogen (N2) to ammonia (NH3) by fixation bacteria. The low quantity demand of molybdenum allows its application on soil and foliar or even by seeds treatments, which is a form of quality aggregation to the seeds by

Cobalt is a component of cobalamin and leghemoglobin synthesis, which is controlling their levels on nodules and avoiding nitrogenase enzyme inactivation;

Nickel can affect directly the presence and quantity of fixation microorganisms because it is a hydrogenase component (Ni-Fe), which can recycle H2 that is

thus this element can be considered essential to N2 fixation [63].

<sup>−</sup>) to nitrite (NO2

<sup>−</sup>) and on the nitrogenase

 *of Ni [64].*

 *of Ni; C1, longitudinal* 

*B1,longitudinal section; B2, approximated image on nodules developed with 0.5 g dm<sup>−</sup><sup>3</sup>*

*section; C2, approximated image on nodules developed with 10 g dm<sup>−</sup><sup>3</sup>*

generated from N reduction and could affect positively or negatively the legume metabolism [64]. Nickel balance on BNF can be seen on fixation nodules where in its absence causes large formation of internal cells according to **Figure 1**.

#### **6. Recent reports about N absence on plants metabolism**

Even though the essentiality had been established for N at higher plants, there are still remained doubts about how the N absence can affect the metabolism. Recently, by modern techniques and sensible equipment, it was possible to determine clearly as N absence affects plant metabolism and production.

The N deficiency exposure of *Olea europaea* plants was described as a significant decrease on chlorophyll a and net photosynthetic rate (**Figure 2**). Photosynthesis is a process that involves light absorption by the photosynthetic pigments present in

#### **Figure 2.**

*Chlorophyll a (Chl a), nitrogen content, and net photosynthetic rate (Amax) in* Olea europaea *plants exposed to nitrogen deficiency [65].*

#### *Nitrogen Fixation*

light-harvesting complexes, being crucial for plant development and largely dependent on the leaf N content, because N composes the chlorophyll molecules [65].

The effects of N deficiency in the leaves of *Oryza sativa* seedlings were verified that the fluorescence parameters were negatively modulated in N-deficient plants [66]. While **Figure 2** presents few modifications until the fifth day in N-deficient plants, when compared with control plants, however as nitrogen deficiency continued, chlorophyll fluorescence of N-deficient plants was significantly impacted, in comparison with control plants.

The decrease in the ratio Fv/Fm of plants under water deficit indicates reduction in the photochemical activity, leading to the inhibition of the photosynthetic rate and the generation of reactive oxygen radicals in the chloroplast, causing damages to PSII components. Additionally, the decline in ETR values of plants under water deficit is due to the deficiency of plastoquinone (PQ ) used in oxidation-reduction reactions.

#### **7. Concluding remarks**

Nitrogen fertilizer consumption follows the increasing demand by food, fiber, and energy production. The quantification of nitrogen inputs and outputs on agricultural system has been a useful and efficient tool to the evaluation of managements, mainly to the tropical agricultural.

Biological fixation is an important nitrogen input to productive systems comprising benefits in economic and environmental concerns, mainly for tropical agriculture; however, the narrow relation among this process and micronutrients and its metabolic routes still needs to be clarified.

Advances of the N fertilization on tropical environment reported at this chapter are focused mainly in an attempt to reduce ammonia volatilization from urea in consequence of its largest use as N source.

Among urease inhibitors used in tropical agriculture, NBPt has been highlighted; however, humic substances have been shown as a future alternative to reduce ammonia volatilization that still requires knowledge about its origin, molecular composition, and environmental questions.

#### **Author details**

Elizeu Monteiro Pereira Junior, Elaine Maria Silva Guedes Lobato\*, Beatriz Martineli Lima, Barbara Rodrigues Quadros, Allan Klynger da Silva Lobato, Izabelle Pereira Andrade and Letícia de Abreu Faria Federal University of Rural Amazônia (UFRA), Paragominas, Brazil

\*Address all correspondence to: elaine.guedes@ufra.edu.br

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

*Advancement of Nitrogen Fertilization on Tropical Environmental DOI: http://dx.doi.org/10.5772/intechopen.90699*

#### **References**

*Nitrogen Fixation*

comparison with control plants.

**7. Concluding remarks**

ments, mainly to the tropical agricultural.

consequence of its largest use as N source.

and its metabolic routes still needs to be clarified.

molecular composition, and environmental questions.

**102**

**Author details**

Elizeu Monteiro Pereira Junior, Elaine Maria Silva Guedes Lobato\*,

Federal University of Rural Amazônia (UFRA), Paragominas, Brazil

\*Address all correspondence to: elaine.guedes@ufra.edu.br

Izabelle Pereira Andrade and Letícia Faria Abreu

provided the original work is properly cited.

Beatriz Martineli Lima, Barbara Rodrigues Quadros, Allan Klynger da Silva Lobato,

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

light-harvesting complexes, being crucial for plant development and largely dependent on the leaf N content, because N composes the chlorophyll molecules [65]. The effects of N deficiency in the leaves of *Oryza sativa* seedlings were verified that the fluorescence parameters were negatively modulated in N-deficient plants [66]. While **Figure 2** presents few modifications until the fifth day in N-deficient plants, when compared with control plants, however as nitrogen deficiency continued, chlorophyll fluorescence of N-deficient plants was significantly impacted, in

The decrease in the ratio Fv/Fm of plants under water deficit indicates reduction in the photochemical activity, leading to the inhibition of the photosynthetic rate and the generation of reactive oxygen radicals in the chloroplast, causing damages to PSII components. Additionally, the decline in ETR values of plants under water deficit is due to the deficiency of plastoquinone (PQ ) used in oxidation-reduction reactions.

Nitrogen fertilizer consumption follows the increasing demand by food, fiber, and energy production. The quantification of nitrogen inputs and outputs on agricultural system has been a useful and efficient tool to the evaluation of manage-

Biological fixation is an important nitrogen input to productive systems comprising benefits in economic and environmental concerns, mainly for tropical agriculture; however, the narrow relation among this process and micronutrients

Advances of the N fertilization on tropical environment reported at this chapter are focused mainly in an attempt to reduce ammonia volatilization from urea in

Among urease inhibitors used in tropical agriculture, NBPt has been highlighted; however, humic substances have been shown as a future alternative to reduce ammonia volatilization that still requires knowledge about its origin,

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[21] Wetsellar R, Ganry F. In: Dommergues YR, Diem HG, editors. Microbiology of Tropical Soils and Plant Productivity. The Hague/Boston/ London: Junk Publishers; 1982

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[25] Malavolta E. Manual de Nutrição Mineral de Plantas. Agronômica Ceres: São Paulo; 2006. 638 p

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[27] Vitti AC, Trivelin PCO, Gava GJC, Franco HCJ, Bologna IR, Faroni CE. Produtividade da cana-de-açúcar relacionada à localização de adubos nitrogenados aplicados sobre os resíduos culturais em canavial sem queima. Revista Brasileira de Ciência do Solo. 2007;**31**:491-498

[28] Cameron KC, Moir HJD. Nitrogen losses from the soil/plant system: A review. The Annals of Applied Biology. 2013;**162**:145-173

[29] Martens DA, Bremmer JM. Soil properties affecting volatilization of ammonia from soils treated with urea. Communications in Soil Science and Plant Analysis. 1989;**20**:1645-1657

[30] Watson CA. The influence of soil properties on the effectiveness of phenylphosphorodiamidate (PPD) in reducing ammonia volatilization from surface applied urea. Nutrient Cycling in Agroecosystems. 1990;**24**:1-10

[31] Bussink DW, Oenema O. Ammonia volatilization from dairy farming systems in temperate areas: A review. Nutrient Cycling in Agroecosystems. 1998;**51**:19-33

[32] Bishop P, Manning M. Urea Volatilization: The Risk Management and Mitigation Strategies. Palmerston North, New Zealand: Fertilizer and Lime Research Centre, Massey University; 2010

[33] Sutton MA, Bleeker A, Howard CM, Bekunda M, Grizzetti B, De Vries W, et al. Our Nutrient World: The Challenge to Produce More Food and Energy with Less Pollution. Edinburgh: Centre for Ecology and Hidrology; 2013

[34] Stafanato JB, Goulart RS, Zonta E, Lima E, Mazur N, Pereira CG, et al. Volatilização de amônia oriunda de ureia pastilhada com micronutrientes em ambiente controlado. Revista Brasileira Ciência do Solo. 2013;**37**:726-732

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[44] Bremner JM, Douglas LA. Inhibition of urease activity in soils. Soil Biology and Biochemistry. 1971;**3**:297-307

[45] Dominguez MJ, SanMartin C, Font M, Palop J, San Francisco J, Urrutia O, et al. Design synthesis and biological evaluation of phosphoramide derivatives as ureases inhibitors. Journal of Agricultural and Food Chemistry.

[46] Watson CJ, Akhonzada NA, Hamilton JTG, Matthews DI. Rate and mode of application of the urease inhibitor N-(n-butyl) thiophosphoric triamide on ammonia volatilization from surface-applied urea. Soil Use and

Management. 2008;**24**:246-253

[47] Ahmed OH, Aminuddin H, Husni MHA. Reducing ammonia loss from urea and improving soilexchangeable ammonium retention through mixing triple superphosphate, humic acid and zeolite. Soil Use and Management. 2006;**22**:315-319

[48] Dong L, Kreylos AL, Yang J, Yuana H, Scowb KM. Humic acids buffer the effects of urea on soil ammonia oxidizers and potential nitrification. Soil Biology and Biochemistry. 2009;**4**:1612-1621

[49] Kasim S, Ahmed OH, Majid NMA, Yusop MK, Jalloh MB. Reduction of ammonia loss by mixing urea with liquid humic and fulvic acids isolated from tropical peat soil. American Journal Agricultural Biology Science.

[50] Canellas LP, Piccolo A, Dobbss LB, Spaccini R, Olivares FL, Zandonadi DB,

et al. Chemical composition and bioactivity properties of size fractions

2009;**4**:18-23

n°. 157; 2017

2008;**56**:3721-3731

*DOI: http://dx.doi.org/10.5772/intechopen.90699*

nitrogênio e produtividade de milho em resposta à aplicação de misturas de ureia com sulfato de amônio ou com gesso agrícola. Revista Brasileira Ciência do

Solo. 2008;**32**:2331-2342

2009;**41**:1270-1280

Solo. 2013;**37**:1057-1063

1940;**5**:238-241

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[37] Nascimento CAC, Vitti GC, Faria LA, Luz PHC, Mendes FL. Ammonia volatilization from coated urea forms. Revista Brasileira Ciência do

[38] Conrad JP. Catalytic activity causing the hydrolysis of urea in soil as influenced by several agronomic factors. Soil Science Society of America Journal.

[39] Kappaun K, Piovesan A, Celia R, Carlini R, Ligabue-Braun R. Ureases: Historical aspects, catalytic and noncatalytic properties – A review. Journal of Advanced Research. 2008;**13**:3-17

[40] Trenkel ME. Slow and Controlled-Release and Stabilized Fertilizers: An Option for Enhancing Nutrient Use Efficiency in Agriculture. Paris: Internacional Fertilizer Industry

[41] Azeem B, Kushaari K, Man ZB, Basit A, Thanh TH. Review on materials and methods to produce controlled release coated urea fertilizer. Journal of Controlled Release. 2014;**181**:11-21

[42] Timilsena YP, Adhikari R, Casey P, Muster T, Gill H, Adhikari B. Enhanced efficiency fertilizers: A review of formulation and nutrient release patterns. Journal of the Science of Food and Agriculture. 2014;**95**:1131-1142

Association; 2010. 167 p

*Advancement of Nitrogen Fertilization on Tropical Environmental DOI: http://dx.doi.org/10.5772/intechopen.90699*

nitrogênio e produtividade de milho em resposta à aplicação de misturas de ureia com sulfato de amônio ou com gesso agrícola. Revista Brasileira Ciência do Solo. 2008;**32**:2331-2342

*Nitrogen Fixation*

quebra de produtividade. Informativo

culturais em canavial sem queima. Revista Brasileira de Ciência do Solo.

[28] Cameron KC, Moir HJD. Nitrogen losses from the soil/plant system: A review. The Annals of Applied Biology.

[29] Martens DA, Bremmer JM. Soil properties affecting volatilization of ammonia from soils treated with urea. Communications in Soil Science and Plant Analysis. 1989;**20**:1645-1657

[30] Watson CA. The influence of soil properties on the effectiveness of phenylphosphorodiamidate (PPD) in reducing ammonia volatilization from surface applied urea. Nutrient Cycling in Agroecosystems. 1990;**24**:1-10

[31] Bussink DW, Oenema O. Ammonia volatilization from dairy farming systems in temperate areas: A review. Nutrient Cycling in Agroecosystems.

[32] Bishop P, Manning M. Urea Volatilization: The Risk Management and Mitigation Strategies. Palmerston North, New Zealand: Fertilizer and Lime Research Centre, Massey

[33] Sutton MA, Bleeker A,

Howard CM, Bekunda M, Grizzetti B, De Vries W, et al. Our Nutrient World: The Challenge to Produce More Food and Energy with Less Pollution. Edinburgh: Centre for Ecology and

[34] Stafanato JB, Goulart RS, Zonta E, Lima E, Mazur N, Pereira CG, et al. Volatilização de amônia oriunda de ureia pastilhada com micronutrientes em ambiente controlado. Revista Brasileira Ciência do Solo. 2013;**37**:726-732

[35] Cabezas WARL, Souza MA. Volatilização de amônia, lixiviação de

2007;**31**:491-498

2013;**162**:145-173

1998;**51**:19-33

University; 2010

Hidrology; 2013

técnico do Núcleo de Sistemas Agrícolas da Embrapa Pesca e Aquicultura n° 14; 2016

[19] Reetz HF. In: Lopes AS, editor. Fertilizantes e o seu uso eficiente. São

[20] Sainju UM. Determination of nitrogen balance in agroecossystems.

Dommergues YR, Diem HG, editors. Microbiology of Tropical Soils and Plant Productivity. The Hague/Boston/

Paulo: ANDA; 2017. 178p

MethodsX. 2017;**4**:199-208

[21] Wetsellar R, Ganry F. In:

London: Junk Publishers; 1982

[22] Guareschi FR, Boddey RM, Alves JR, Sarkis LF, Martins MR, Jantalia CP, et al. Balanço de nitrogênio, fósfoto e potássio na agricultura da América Latina e o Caribe. Revista Terra Latinoamericana. 2019;**37**:105-119

[23] De Datta SK. Principles and

[24] Cantarella H, Mattos Júnior D, Quaggio JA, Rigolin AT. Fruit yield of Valencia sweet orange fertilized with different N sources and the loss of applied N. Nutrient Cycling in Agroecosystems. 2003;**67**:215-223

[25] Malavolta E. Manual de Nutrição Mineral de Plantas. Agronômica Ceres:

[26] Cantarella H. Nitrogênio. In: Novais RF, Alvarez VVH, Barros NF, Fontes RLF, Cantarutti RB, Neves JCL, editors. Fertilidade do Solo. Viçosa:

[27] Vitti AC, Trivelin PCO, Gava GJC, Franco HCJ, Bologna IR, Faroni CE. Produtividade da cana-de-açúcar relacionada à localização de adubos nitrogenados aplicados sobre os resíduos

John Wiley; 1981. 618 p

São Paulo; 2006. 638 p

SBCS; 2007. pp. 422-423

Practices of Rice Production. New York:

**104**

[36] Zaman M, Saggar S, Blennerhassett JD, Singh J. Effect of urease and nitrification inhibitors on N transformation, gaseous emissions of ammonia and nitrous oxide, pasture yield and N uptake in grazed pasture system. Soil Biology and Biochemistry. 2009;**41**:1270-1280

[37] Nascimento CAC, Vitti GC, Faria LA, Luz PHC, Mendes FL. Ammonia volatilization from coated urea forms. Revista Brasileira Ciência do Solo. 2013;**37**:1057-1063

[38] Conrad JP. Catalytic activity causing the hydrolysis of urea in soil as influenced by several agronomic factors. Soil Science Society of America Journal. 1940;**5**:238-241

[39] Kappaun K, Piovesan A, Celia R, Carlini R, Ligabue-Braun R. Ureases: Historical aspects, catalytic and noncatalytic properties – A review. Journal of Advanced Research. 2008;**13**:3-17

[40] Trenkel ME. Slow and Controlled-Release and Stabilized Fertilizers: An Option for Enhancing Nutrient Use Efficiency in Agriculture. Paris: Internacional Fertilizer Industry Association; 2010. 167 p

[41] Azeem B, Kushaari K, Man ZB, Basit A, Thanh TH. Review on materials and methods to produce controlled release coated urea fertilizer. Journal of Controlled Release. 2014;**181**:11-21

[42] Timilsena YP, Adhikari R, Casey P, Muster T, Gill H, Adhikari B. Enhanced efficiency fertilizers: A review of formulation and nutrient release patterns. Journal of the Science of Food and Agriculture. 2014;**95**:1131-1142

[43] Guelfi D. Fertilizantes Nitrogenados Estabilizados de liberação lenta ou controlada. Informações Agronômicas n°. 157; 2017

[44] Bremner JM, Douglas LA. Inhibition of urease activity in soils. Soil Biology and Biochemistry. 1971;**3**:297-307

[45] Dominguez MJ, SanMartin C, Font M, Palop J, San Francisco J, Urrutia O, et al. Design synthesis and biological evaluation of phosphoramide derivatives as ureases inhibitors. Journal of Agricultural and Food Chemistry. 2008;**56**:3721-3731

[46] Watson CJ, Akhonzada NA, Hamilton JTG, Matthews DI. Rate and mode of application of the urease inhibitor N-(n-butyl) thiophosphoric triamide on ammonia volatilization from surface-applied urea. Soil Use and Management. 2008;**24**:246-253

[47] Ahmed OH, Aminuddin H, Husni MHA. Reducing ammonia loss from urea and improving soilexchangeable ammonium retention through mixing triple superphosphate, humic acid and zeolite. Soil Use and Management. 2006;**22**:315-319

[48] Dong L, Kreylos AL, Yang J, Yuana H, Scowb KM. Humic acids buffer the effects of urea on soil ammonia oxidizers and potential nitrification. Soil Biology and Biochemistry. 2009;**4**:1612-1621

[49] Kasim S, Ahmed OH, Majid NMA, Yusop MK, Jalloh MB. Reduction of ammonia loss by mixing urea with liquid humic and fulvic acids isolated from tropical peat soil. American Journal Agricultural Biology Science. 2009;**4**:18-23

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Science. 2016;**8**(7):1358. DOI: 10.3389/

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**106**

**Chapter 7**

**Abstract**

**1. Introduction**

**109**

Soybean

Comprehensive Account of

Inoculation and Coinoculation in

*Muhammad Jamil Khan, Rafia Younas, Abida Saleem,*

*Mumtaz Khan, Qudratullah Khan and Rehan Ahmed*

technical and economic aspects of coinoculation in soybean.

**Keywords:** legumes, nodulation, BNF, phytohormones, mixed inoculation

Better plant growth is ensured by the balanced availability of essential nutrients

in soil. Each nutrient has its own function and is required in different amount depending on the plant demand. Nitrogen (N), one of the most essential macronutrients, is routinely applied through chemical fertilizer as most field crops require large amounts of it. Nitrogen, the fifth most abundant element in the universe, was first discovered in 1772 by a Scottish physician, Daniel Rutherford. Due to its essentiality for survival of life on earth, it was called as "azote," meaning "without life," by Antoine Lavoisier about 200 years. Nitrogen is essential for the sustenance of life on this planet as it serves as building block for the synthesis of proteins. The inevitable role of N is well acknowledged in several biochemical processes such as cell division, growth promotion, and photosynthesis, as part of vitamins and carbohydrates and energy reactions in the plant body [1, 2]. Deficiency of N in plants is

This chapter elaborates dependency of leguminous plants on rhizobia to carry out dynamic process of nitrogen fixation. Soybean, an extensively grown leguminous crop with 30% share in world's vegetable oil, is taken into account to understand its symbiotic relationship with plant growth-promoting rhizobacteria (PGPRs). This chapter narrates colonization of PGPRs on soybean roots and single and mixed inoculation and coinoculation of certain strains of specialized bacteria with rhizobia. PGPRs' coinoculation seemed more effective than mono-inoculation and is discussed in Ref. to nodulation rate. Moreover, dynamic linear models for quantification of leguminous biological nitrogen fixation (BNF) are reviewed. This chapter further uncoils the relevance of foliar application to the release of phytohormones by PGPRs, resulting in situ biosynthesis of active metabolites in phyllosphere. Inoculation of phytohormones is compared to their exogenous application for nodule organogenesis. Finally, the influence of coinoculation on enhanced micronutrient bioavailability is relayed. The chapter is concluded with

#### **Chapter 7**
