Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen (15 N) and AquaCrop

*Mawhoub Amirouche, Dalila Smadhi and Lakhdar Zella*

#### **Abstract**

The present study is highlighted through an experiment carried out over two consecutive years 2014–2016, in the sub humid region of Algiers. The methodology adopted concerns the variation of optimal nitrogen doses and their effects on the evolution of lettuce (*Lactuca sativa* L.) cultivation, whose socio-economic impact is proven, using isotopic nitrogen (15 N) and the AquaCrop model. The experimental design adopted is of the complete randomized block type, with four (04) levels: 0 (control), 60, 120, and 180 kg N/ha with four (04) replicates. The results obtained showed that the 120 kg N/ha dose is the efficient dose to cover the nitrogen requirements of lettuce with an efficiency of 74.48%. The accuracy of the model in calibration was tested using the following statistical indicators: R2, nRMSE, and d, which are, respectively, 0.64 < R2 < 0.81; 18 < nRMSE <46.3 and 0.78 < d < 0.94 for canopy coverage and 0.92 < R2 < 0.98; 21.6 < nRMSE <34.5 and 0.91 < d < 0.96 for dry biomass. The AquaCrop model could be recommended as a practical tool to better manage agricultural practices including fertilization.

**Keywords:** lettuce, AquaCrop, isotopic nitrogen 15 N, nitrogen use efficiency, fertility stress

#### **1. Introduction**

In the coming years, agricultural production will have to face a double challenge, meeting the growing needs of the world's population while preserving the environment and natural resources. According to [1], the world's current population of about 6.3 billion people will reach nearly 8.6 billion in 2030. Agricultural production will then have to be significantly higher. This will be achieved by increasing yields. This has been achieved mainly through varietal improvement and associated cultivation techniques, including nitrogen fertilization.

In Algeria, 20% of the agricultural potential is located in the north of the country, which is characterized by poorly fertile soils. These soils are low in nutrients and have a very low rate of organic matter. Fertilization has remained archaic in the country. According to [2], in Algeria, the use of fertilizers in agriculture is not under control, despite the efforts made by farmers in charge of the cereal intensification program and potato farmers.

According to [3], fertilizers are applied in the absence of technical standards, neglecting the initial soil content; consequently, inputs are often poorly fractioned, leading to waste, which is a source of soil and water pollution. As several researchers have shown in their work on Algerian soils. In this context, [4, 5] conducted trials in the same semi-arid climate, respectively, on durum wheat and barley seed production, obtained maximum yields with similar rates (150 kg N/ha). These yields reached the respective values of 33.82 and 33.25 q/ha, i.e. gains of 11.52 and 9.76 q/ha. Halilat [6] showed that the interaction of potassium (P) and nitrogen (N) fertilization significantly affects wheat grain yield in the Saharan zone. The maximum yield reached 6.780 Mt./ha with the N250 P180 dose. With regard to nitrogenous fertilization, the observation highlights the need to promote adapted and balanced fertilization. Since urea is the most widely used nitrogen fertilizer in the world [7], it is crucial to assess the nitrogen use efficiency (NUE) by crops, since it is always aimed at achieving higher yields with a minimum application of fertilizer. This indicator (NUE) has been widely studied by several researchers around the world on various crops, including cereals, e.g. rice [8], maize [9], durum wheat [10, 11]; leafy vegetables, e.g. lettuce [12–14], spinach [15], cabbage [16] and vegetable crops, e.g. Potato [17, 18]; beans [19], tomato [20, 21].

In this perspective, this study uses the isotope approach 15 N to evaluate the nitrogen use efficiency. This new method, used by [22], highlights 15 N isotopic nitrogen, which is the most commonly used stable isotope in agriculture-related studies. It is the direct way to measure nitrogen uptake by applied fertilizer, and the most reliable way to monitor the flow and fate of nitrogen in the soil–plant system [23, 24]. To highlight the monitoring of this system, the chosen plant material is lettuce (*Lactuca sativa* L.), due to its short growing cycle. But also, because of the socio-economic impact that is beginning to dominate, at the national level. It is a source of wealth and income for producers. The search for decision support tools is essential in order to master agricultural practices and to plan for a sustainable agriculture that respects the environment. In this respect, the AquaCrop model, designed by the FAO, has been chosen as a decision support tool. The objective of this study is essentially oriented toward the search for optimum doses of nitrogenous fertilizers with the aim of contributing to the production of technical references for the efficient use of fertilizers.

#### **2. Materials and methods**

#### **2.1 Study site**

The study was conducted at the National Institute of Agronomic Research of Algiers (36°68′ N and 3°1′ E, at an altitude of 18 m), located south-west of Algiers in the eastern part of the Mitidja (**Figure 1**).

#### **2.2 Climatic and soil data**

Climatic conditions in the study area are characterized by pronounced seasonal variations with mild, wet winters and hot, dry summers. The meteorological data used are from the automatic weather station installed in the field. The measurements taken at daily time steps are: minimum and maximum temperatures (°C), rainfall (mm), wind speed (m/s) at 2 m above ground level, solar radiation (W/m2 ) and relative humidity (%). The reference evapotranspiration (ET0) was calculated according to the FAO Penman-Monteith method [25]. A soil profile was carried out

**101**

*Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen…*

over a depth of one meter, comprising three horizons. Soil samples were taken from

The crop taken into consideration is variety lettuce, stubborn from Nîmes, belonging to the lettuce to be applesauce class, which is eaten young, before it goes to seed. Lettuce seeds were sown in the honeycomb plates for 19–25 days in the nursery before being transplanted. The young lettuce plants were transplanted at

The experiment was carried out in the open field using a complete randomized block experimental design with four levels of nitrogen, namely: T1 (0 N kg/ha), T2 (60 N kg/ha), T3 (120 N kg/ha) and T4 (180 N kg/ha) arranged in four blocks. Each block has four sub-plots. Each micro plot is 6 m long and 3 m wide, giving a total

consecutive years (2014–2015) and (2015–2016). Isotopic nitrogen was used only in the first year because of its high cost. The amounts of nitrogen used were distributed along the crop development cycle, namely: 10% at 15 days after transplanting (DAT), 30% at 40 DAT, 40% at 60 DAT and 20% at 75 DAT. The growing season is from January to April for both companions, coinciding with the winter season,

The parameters measured in the field are essentially the above-ground biomass (B), which represents a parameter that best allows verification of fertilizer efficiency in lettuce where the growth of the above-ground part is a determining factor in agricultural value [26]. Every 10 days, samples of 6 plants/subplot are taken and brought back to the laboratory where they are dried in the open air for 24 h and then in an oven for 48 h at 70 C°. In addition to this, the evolution of the green canopy (CC) cover is monitored by reference to photos taken vertically at a height of 1.8 m above the crop, using a photometric device. The photos were analyzed using ARCgis 10.1 software using the supervised classification by maximum likelihood method (**Figure 2**). Harvesting was done when the apples were tightly packed and full for

was used for the 15 N. The trial was repeated for two

each horizon with an auger for analysis physico-chemical.

the 3–4 leaf stage onto well plowed soil in the field.

, of which 4.5 m2

during which irrigation is not necessary.

**2.5 Measured parameters**

each subplot of 1 m × 1 m.

**2.3 Crop data**

*Location of the study area.*

**Figure 1.**

area of 18 m2

**2.4 Experimental protocol**

*DOI: http://dx.doi.org/10.5772/intechopen.93741*

*Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen… DOI: http://dx.doi.org/10.5772/intechopen.93741*

**Figure 1.** *Location of the study area.*

over a depth of one meter, comprising three horizons. Soil samples were taken from each horizon with an auger for analysis physico-chemical.

#### **2.3 Crop data**

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

[17, 18]; beans [19], tomato [20, 21].

the efficient use of fertilizers.

**2. Materials and methods**

**2.2 Climatic and soil data**

the eastern part of the Mitidja (**Figure 1**).

**2.1 Study site**

According to [3], fertilizers are applied in the absence of technical standards, neglecting the initial soil content; consequently, inputs are often poorly fractioned, leading to waste, which is a source of soil and water pollution. As several researchers have shown in their work on Algerian soils. In this context, [4, 5] conducted trials in the same semi-arid climate, respectively, on durum wheat and barley seed production, obtained maximum yields with similar rates (150 kg N/ha). These yields reached the respective values of 33.82 and 33.25 q/ha, i.e. gains of 11.52 and 9.76 q/ha. Halilat [6] showed that the interaction of potassium (P) and nitrogen (N) fertilization significantly affects wheat grain yield in the Saharan zone. The maximum yield reached 6.780 Mt./ha with the N250 P180 dose. With regard to nitrogenous fertilization, the observation highlights the need to promote adapted and balanced fertilization. Since urea is the most widely used nitrogen fertilizer in the world [7], it is crucial to assess the nitrogen use efficiency (NUE) by crops, since it is always aimed at achieving higher yields with a minimum application of fertilizer. This indicator (NUE) has been widely studied by several researchers around the world on various crops, including cereals, e.g. rice [8], maize [9], durum wheat [10, 11]; leafy vegetables, e.g. lettuce [12–14], spinach [15], cabbage [16] and vegetable crops, e.g. Potato

In this perspective, this study uses the isotope approach 15 N to evaluate the nitrogen use efficiency. This new method, used by [22], highlights 15 N isotopic nitrogen, which is the most commonly used stable isotope in agriculture-related studies. It is the direct way to measure nitrogen uptake by applied fertilizer, and the most reliable way to monitor the flow and fate of nitrogen in the soil–plant system [23, 24]. To highlight the monitoring of this system, the chosen plant material is lettuce (*Lactuca sativa* L.), due to its short growing cycle. But also, because of the socio-economic impact that is beginning to dominate, at the national level. It is a source of wealth and income for producers. The search for decision support tools is essential in order to master agricultural practices and to plan for a sustainable agriculture that respects the environment. In this respect, the AquaCrop model, designed by the FAO, has been chosen as a decision support tool. The objective of this study is essentially oriented toward the search for optimum doses of nitrogenous fertilizers with the aim of contributing to the production of technical references for

The study was conducted at the National Institute of Agronomic Research of Algiers (36°68′ N and 3°1′ E, at an altitude of 18 m), located south-west of Algiers in

Climatic conditions in the study area are characterized by pronounced seasonal variations with mild, wet winters and hot, dry summers. The meteorological data used are from the automatic weather station installed in the field. The measurements taken at daily time steps are: minimum and maximum temperatures (°C), rainfall (mm), wind speed (m/s) at 2 m above ground level, solar radiation (W/m2

and relative humidity (%). The reference evapotranspiration (ET0) was calculated according to the FAO Penman-Monteith method [25]. A soil profile was carried out

**100**

The crop taken into consideration is variety lettuce, stubborn from Nîmes, belonging to the lettuce to be applesauce class, which is eaten young, before it goes to seed. Lettuce seeds were sown in the honeycomb plates for 19–25 days in the nursery before being transplanted. The young lettuce plants were transplanted at the 3–4 leaf stage onto well plowed soil in the field.

#### **2.4 Experimental protocol**

The experiment was carried out in the open field using a complete randomized block experimental design with four levels of nitrogen, namely: T1 (0 N kg/ha), T2 (60 N kg/ha), T3 (120 N kg/ha) and T4 (180 N kg/ha) arranged in four blocks. Each block has four sub-plots. Each micro plot is 6 m long and 3 m wide, giving a total area of 18 m2 , of which 4.5 m2 was used for the 15 N. The trial was repeated for two consecutive years (2014–2015) and (2015–2016). Isotopic nitrogen was used only in the first year because of its high cost. The amounts of nitrogen used were distributed along the crop development cycle, namely: 10% at 15 days after transplanting (DAT), 30% at 40 DAT, 40% at 60 DAT and 20% at 75 DAT. The growing season is from January to April for both companions, coinciding with the winter season, during which irrigation is not necessary.

#### **2.5 Measured parameters**

The parameters measured in the field are essentially the above-ground biomass (B), which represents a parameter that best allows verification of fertilizer efficiency in lettuce where the growth of the above-ground part is a determining factor in agricultural value [26]. Every 10 days, samples of 6 plants/subplot are taken and brought back to the laboratory where they are dried in the open air for 24 h and then in an oven for 48 h at 70 C°. In addition to this, the evolution of the green canopy (CC) cover is monitored by reference to photos taken vertically at a height of 1.8 m above the crop, using a photometric device. The photos were analyzed using ARCgis 10.1 software using the supervised classification by maximum likelihood method (**Figure 2**). Harvesting was done when the apples were tightly packed and full for each subplot of 1 m × 1 m.

)

**Figure 2.** *Analyses of the fraction of the green canopy for the growth stage.*

To determine the isotopic composition of lettuce plants, lettuce heads receivingan15N were divided into two parts (roots and leaves). Fresh weight was assessed for all parts of the crop. The samples were dried at 70°C for 24 hours, weighed for dry weight determination, ground into a fine powder using a 0.3 mm sieve and homogenized for total nitrogen and excess N15. The isotopic analysis of the lettuce culture samples was carried out at the National Centre for Energy, Science and Nuclear Techniques (CNESTEN-Morocco).

The quantification of fertilizer nitrogen was measured on the basis of the isotope dilution method from fertilizer nitrogen and the rate of nitrogen fertilizer applied, according to the following equation defined by [22]:

$$\text{96 Ferrilizer N utilization} = \frac{\text{Fertlizer N yield}}{\text{Rate of Naprlization}} \ast \mathbf{100} \tag{1}$$

#### **2.6 Description and evaluation of the data by AquaCrop**

AquaCrop requires five important components to be functional: climate, with its thermal regime, rainfall, evaporative demand (ETP) and carbon dioxide concentration; then crop characteristics, including development, growth and yield formation processes (**Table 1**); then soil, with its hydraulic characteristics (hydraulic conductivity at saturation, moisture at saturation, field capacity and permanent wilting point); and finally management practices, which are divided into two categories: plot management and irrigation practice management; and finally initial conditions.

#### **2.7 Model calibration for soil fertility stress**

Calibration of the model to fertility stress requires coverage of the green canopy (CC) and biomass production (B), recorded on the fertility stressed plot 'stressed plot' and the unstressed plot 'reference plot' (**Table 2**). The soil fertility stress in the AquaCrop model is given as follows:

$$\text{Stress} = \mathbf{100} \left( \mathbf{1} - \mathbf{B}\_{nl} \right) \tag{2}$$

**103**

**Table 2.**

**Table 1.**

*Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen…*

**Description Units 2015–16 Source**

Base temperature C° 7 Calibrated Upper temperature C° 30 Calibrated Upper threshold for canopy expansion, Pexp,upper — 0.25 Simulated Lower threshold for canopy expansion, Pexp,lower — 0.55 Simulated

Upper threshold for stomatal closure, Psto,upper - 0.50 Calibrated Shape factor for the stress coefficient for stomatal closure — 3 Calibrated Water productivity (WP) g m−2 19 Calibrated Reference harvest index (HIo) % 95 Measured Crop coefficient when canopy is complete — 0.85 Simulated

Number of plants per m2 Plant m−2 15 Measured CC0 % 2.25 Simulated Maximum canopy cover CCx % 81 Measured

Time from transplantation to emergence Days 7 Observed Time from transplantation to senescence Days 80 Observed Time from transplantation to maximum (CCx) Days 50 Observed Time from transplantation to maturity Days 95 Observed Minimum effective rooting depth m 0.20 Measured Maximum effective rooting depth m 0.40 Measured Transplantation time at maximum depth of rooting Days 55 Observed

Canopy growth coefficient (CGC) % days−1 14.30 Simulated Canopy decline coefficient (CDC) % days−1 8.0 Simulated

**Treatments Brel (%) CCx under fertility level (%) Canopy Decline (-)** T1 51 51 Strong T2 73 55 Medium T3 100 61 Little T4 100 58 Little

plant−1

**Non conservative parameters** 2015–16

Date of transplantation 11/01/16 Harvest date 14/04/2016

Canopy size of the transplanted seedling cm2

*Input culture parameters to calibrate the AquaCrop model.*

*Input data to calibrate the AquaCrop model for soil fertility stress.*

— 3 Calibrated

15 Measured

*DOI: http://dx.doi.org/10.5772/intechopen.93741*

Shape factor for the stress coefficient for canopy

**Conservative crop parameters**

expansion

Where: Brel is the ratio of total dry above-ground biomass at the end of the growing season in the reference plot (Bref) to that under stress (Bstress). Soil fertility

*Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen… DOI: http://dx.doi.org/10.5772/intechopen.93741*


#### **Table 1.**

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

Nuclear Techniques (CNESTEN-Morocco).

*Analyses of the fraction of the green canopy for the growth stage.*

**Figure 2.**

according to the following equation defined by [22]:

**2.6 Description and evaluation of the data by AquaCrop**

**2.7 Model calibration for soil fertility stress**

AquaCrop model is given as follows:

To determine the isotopic composition of lettuce plants, lettuce heads receivingan15N were divided into two parts (roots and leaves). Fresh weight was assessed for all parts of the crop. The samples were dried at 70°C for 24 hours, weighed for dry weight determination, ground into a fine powder using a 0.3 mm sieve and homogenized for total nitrogen and excess N15. The isotopic analysis of the lettuce culture samples was carried out at the National Centre for Energy, Science and

The quantification of fertilizer nitrogen was measured on the basis of the isotope dilution method from fertilizer nitrogen and the rate of nitrogen fertilizer applied,

AquaCrop requires five important components to be functional: climate, with its thermal regime, rainfall, evaporative demand (ETP) and carbon dioxide concentration; then crop characteristics, including development, growth and yield formation processes (**Table 1**); then soil, with its hydraulic characteristics (hydraulic conductivity at saturation, moisture at saturation, field capacity and permanent wilting point); and finally management practices, which are divided into two categories: plot management and irrigation practice management; and finally initial conditions.

Calibration of the model to fertility stress requires coverage of the green canopy (CC) and biomass production (B), recorded on the fertility stressed plot 'stressed plot' and the unstressed plot 'reference plot' (**Table 2**). The soil fertility stress in the

Where: Brel is the ratio of total dry above-ground biomass at the end of the growing season in the reference plot (Bref) to that under stress (Bstress). Soil fertility

*RateofNapplication <sup>=</sup>* (1)

Stress 100 1 B = − ( *rel*) (2)

% Fertilizer N utilization <sup>∗</sup><sup>100</sup> *Fertilizer N yield*

**102**

*Input culture parameters to calibrate the AquaCrop model.*


#### **Table 2.**

*Input data to calibrate the AquaCrop model for soil fertility stress.*

affects water productivity (WP), canopy growth coefficient (CGC), maximum cover (CCx) and canopy senescence.

The evolution of canopy cover, dry above-ground biomass and yield were taken into account in the evaluation of the AquaCrop model, while using the following statistical indicators: the coefficient of determination (R<sup>2</sup> ) of the linear fit, the square root of the normalized root mean square error (nRMSE) and the Willmott's agreement index (d).

#### **3. Results and discussions**

#### **3.1 Analysis of climate data**

Variations in rainfall and ETP are shown in **Figure 3**, which illustrates the rainfall distribution during the two years of experience 2014–2015 and 2015–2016. The cumulative rainfall received between September and August is, respectively, of the order of 552 and 551 mm. Those corresponding to the experimental seasons (January to April), they are close to the averages of 211.4 and 303.4 mm. The corresponding potential annual evapotranspiration is of the order of 744.3 and 782.6 mm. Those corresponding to the growing seasons are, respectively, 195.4 and 196.5 mm.

#### **3.2 Physical and chemical characteristics of the soil**

The study site is characterized by deep and heavy soils with high clay content. Soil analysis revealed the existence of 3 horizons with a silty-clay texture with high clay rates increasing with soil depth. At profiles of 0–25 cm, 25–55 cm and beyond 55 cm depth, these rates are 43, 49, and 52%, respectively. The pH of the station soils is generally slightly basic at 7.8, CEC varies between 17.9 and 15 meq/100 g and total limestone has a rate between 7.9 and 7.8%. The organic matter rate is 1.57% on the surface and 0.49% at depth.

#### **3.3 Effect of fertilization on dry above-ground biomass**

**Figure 4** shows the evolution of the nitrogen doses applied at different phenological stages of the plant. This evolution is supported by the analysis of variance, which showed a very highly significant effect (p < 0.001), of the dry biomass, in relation to the increase in the doses of nitrogen supplied. A maximum of dry biomass is reached at the dose of 120 kg N/ha. Above this level, the increase in nitrogen rate is not significant. This result is consistent with that of [27], which showed that fertilization at high doses leads to a decrease in above-ground biomass. This is the case in the first year (2014–2015).

#### **3.4 Effect of fertilization on yields**

**Figure 5** shows lettuce yields as a function of applied nitrogen rates. In fact, the graph shows that, during the two experimental campaigns, the highest lettuce yields (55.24 and 57.96 t/ha) were obtained by applying the 120 and 180 kg/ha rates. These doses are very highly significant (p < 0.001) compared to those obtained (30.19 and 45.49 t/ha) by applying the minimum doses of less than 60 kg/ha. This result is consistent with those of [28–30], who reported that increasing the N level from 0 to 120 kg N/ha had a positive effect on lettuce production. Nevertheless, in detail, the T4 treatment from the 2014–2015 trial shows a relatively lower yield of 50.25 t/ha

**105**

**Figure 3.**

**Figure 4.**

*seasons.*

*Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen…*

*Precipitation, potential evapotranspiration (ETP) on a monthly scale for test years 2014–2015 and 2015–2016.*

compared to the T3 treatment (54.25 t/ha) from the same year. The difference, evaluated at 3.08 t/ha, can be explained by the toxicity of the plants or by the nonattraction of nitrogen by the plants resulting from the consumption of excess nitrogen fertilizer, as pointed out by [31]. The response of lettuce for yields is considerably higher in 2016

*Effect of different levels of fertilization on the evolution of dry above-ground biomass for the two growing* 

Nitrogen Use Efficiency (NUE) is an important indicator in the application of nitrogen fertilizers. Achieving a higher NUE always becomes a priority in agriculture [8]. In this context, **Figure 6** illustrates the variation in the percentage of NUE as a function of defined thresholds. For rates ranging from 60, 120 to 180 kg N/ha, the NUE varies from 65.42, 74.49 to 68.38%, respectively. The NUE decreased from 74.49% to 68.38% by increasing the rate from 120 to 180 kg N/ha. These results are similar to those reported by [32, 33]. The 120 kg N/ha rate provides the best efficiencies. This means that 74.48% of the fertilizer applied is consumed by the lettuce crop. The remaining 25.52% of N is either in the soil or lost through leaching. Lettuce is a short-cycle crop, making the best use of available nitrogen, as reported by [34].

than in 2015. This result is related to the higher rainfall amounts.

**3.5 Nitrogen use efficiency**

*DOI: http://dx.doi.org/10.5772/intechopen.93741*

*Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen… DOI: http://dx.doi.org/10.5772/intechopen.93741*

**Figure 3.** *Precipitation, potential evapotranspiration (ETP) on a monthly scale for test years 2014–2015 and 2015–2016.*

**Figure 4.**

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

statistical indicators: the coefficient of determination (R<sup>2</sup>

**3.2 Physical and chemical characteristics of the soil**

**3.3 Effect of fertilization on dry above-ground biomass**

cover (CCx) and canopy senescence.

agreement index (d).

196.5 mm.

**3. Results and discussions**

**3.1 Analysis of climate data**

the surface and 0.49% at depth.

case in the first year (2014–2015).

**3.4 Effect of fertilization on yields**

affects water productivity (WP), canopy growth coefficient (CGC), maximum

The evolution of canopy cover, dry above-ground biomass and yield were taken into account in the evaluation of the AquaCrop model, while using the following

square root of the normalized root mean square error (nRMSE) and the Willmott's

Variations in rainfall and ETP are shown in **Figure 3**, which illustrates the rainfall distribution during the two years of experience 2014–2015 and 2015–2016. The cumulative rainfall received between September and August is, respectively, of the order of 552 and 551 mm. Those corresponding to the experimental seasons (January to April), they are close to the averages of 211.4 and 303.4 mm. The corresponding potential annual evapotranspiration is of the order of 744.3 and 782.6 mm. Those corresponding to the growing seasons are, respectively, 195.4 and

The study site is characterized by deep and heavy soils with high clay content. Soil analysis revealed the existence of 3 horizons with a silty-clay texture with high clay rates increasing with soil depth. At profiles of 0–25 cm, 25–55 cm and beyond 55 cm depth, these rates are 43, 49, and 52%, respectively. The pH of the station soils is generally slightly basic at 7.8, CEC varies between 17.9 and 15 meq/100 g and total limestone has a rate between 7.9 and 7.8%. The organic matter rate is 1.57% on

**Figure 4** shows the evolution of the nitrogen doses applied at different phenological stages of the plant. This evolution is supported by the analysis of variance, which showed a very highly significant effect (p < 0.001), of the dry biomass, in relation to the increase in the doses of nitrogen supplied. A maximum of dry biomass is reached at the dose of 120 kg N/ha. Above this level, the increase in nitrogen rate is not significant. This result is consistent with that of [27], which showed that fertilization at high doses leads to a decrease in above-ground biomass. This is the

**Figure 5** shows lettuce yields as a function of applied nitrogen rates. In fact, the graph shows that, during the two experimental campaigns, the highest lettuce yields (55.24 and 57.96 t/ha) were obtained by applying the 120 and 180 kg/ha rates. These doses are very highly significant (p < 0.001) compared to those obtained (30.19 and 45.49 t/ha) by applying the minimum doses of less than 60 kg/ha. This result is consistent with those of [28–30], who reported that increasing the N level from 0 to 120 kg N/ha had a positive effect on lettuce production. Nevertheless, in detail, the T4 treatment from the 2014–2015 trial shows a relatively lower yield of 50.25 t/ha

) of the linear fit, the

**104**

*Effect of different levels of fertilization on the evolution of dry above-ground biomass for the two growing seasons.*

compared to the T3 treatment (54.25 t/ha) from the same year. The difference, evaluated at 3.08 t/ha, can be explained by the toxicity of the plants or by the nonattraction of nitrogen by the plants resulting from the consumption of excess nitrogen fertilizer, as pointed out by [31]. The response of lettuce for yields is considerably higher in 2016 than in 2015. This result is related to the higher rainfall amounts.

#### **3.5 Nitrogen use efficiency**

Nitrogen Use Efficiency (NUE) is an important indicator in the application of nitrogen fertilizers. Achieving a higher NUE always becomes a priority in agriculture [8]. In this context, **Figure 6** illustrates the variation in the percentage of NUE as a function of defined thresholds. For rates ranging from 60, 120 to 180 kg N/ha, the NUE varies from 65.42, 74.49 to 68.38%, respectively. The NUE decreased from 74.49% to 68.38% by increasing the rate from 120 to 180 kg N/ha. These results are similar to those reported by [32, 33]. The 120 kg N/ha rate provides the best efficiencies. This means that 74.48% of the fertilizer applied is consumed by the lettuce crop. The remaining 25.52% of N is either in the soil or lost through leaching. Lettuce is a short-cycle crop, making the best use of available nitrogen, as reported by [34].

**Figure 5.** *Effect of different levels of fertilization on yield for the two growing seasons.*

#### **Figure 6.**

*Variation in nitrogen use efficiency by lettuce, expressed as a percentage (%).*

#### **3.6 Effect of fertilization on water productivity**

**Figure 7** shows the variation in water productivity (WP), soil evaporation (Es) and transpiration (Tr) of the lettuce crop under different levels of fertilization. This variation is supported by the analysis of variance, which showed a very highly significant effect (p < 0.001) of these parameters (WP, Es and Tr), in relation to the increase in the doses of nitrogen applied. The maximum values of WP and Tr are reached at the dose of 120 kg N/ha, for the two companions 2014–2015 (WP = 8.95 kg/m3; Tr =51.4 mm) and 2015–2016 (9.57 kg/m3; Tr = 55.80 mm). Above this level, the increase in the nitrogen rate is not significant.

#### **3.7 Calibration of the AquaCrop model**

#### *3.7.1 Canopy cover and dry biomass*

Experimental results of yield, canopy cover and dry above-ground biomass under different levels of fertilization are presented in **Table 3**. The AquaCrop model (V. 6.1) was calibrated using the crop data set obtained from the T3 treatment (120 kg N/ha). The lowest dry yield and dry aboveground biomass observed were 4.021 t/ha and 4.125 t/ha under the T1 treatment (0 kg N/ha), and the highest were 8.854 t/ha and 9.320 t/ha under the T3 treatment (120 kg N/ha), respectively.

**107**

R2

*3.7.2 Yield*

**Figure 7.**

**Table 3.**

*2015–2016.*

*Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen…*

is satisfactory with 0.64 < R2 < 0.81, 18 < nRMSE > 46.3 and 0.78 < d < 0.94;

*Biomass calibration results, yield, and maximum canopy cover under different levels of fertilization in* 

**Figure 8** shows the comparison between simulated and observed canopy cover (CC) and dry above-ground biomass (B) for the calibration period (2015–2016). This figure shows that there is a close correspondence between observed and simulated CC and B. It is also important to note that the AquaCrop model correctly simulates CC from seeding to the maximum growth phase at which CCx is reached. This observation has been reported in several studies [35–37]. From **Figure 8**, it is clear that both parameters (CC) and B were overestimated by the AquaCrop model. In a recent study [38], it was shown that the AquaCrop model overestimated the cabbage canopy under different irrigation regimes. Nikolaus [39] also noted a slight (10%) but systematic overestimation of the amount of rice biomass conducted

**Treatments Biomass (t/ha) Dry yield (t/ha) CCx (%)**

T1 4.125 4.785 4.021 4.546 (6.70) 51 44.80 (1.68) T2 5.872 6.806 5.234 5.785 (11.68) 55 54.10 (5.76 T3 7.969 9.320 7.834 8.854 (8.27) 61 63.90 (2.67) T4 7.788 9.100 7.626 8.645 (8.40) 58 63.70 (3.55)

**Obs Sim Obs Sim SD (±%) Obs Sim SD (±%)**

Observed and simulated lettuce yields are shown in **Figure 9**. The observed yields for treatments T1, T2, T3 and T4 are, respectively, 4.214; 5.187; 6.942 and 6.214 t/ha, while the simulated yields are 4.897; 5.981; 7.414 and 6.987 for the trial

the yields observed and simulated under the four treatments for the trial period (2015–2016) are of the order of 4.021; 5.234; 7834 and 7.626 t/ha, while those simulated are of the order of 4.546; 5.785; 8.854 and 8.645, with a correlation coefficient

= 0.99. Analysis of statistical tests and linear regression indicated that the values

= 0.92. On the other hand,

0.92 < R2 < 0.94, 21.6 < nRMSE <34.5, 0.91 < d < 0.96 (**Table 4**).

*Variation in water productivity of lettuce under different levels of fertilization.*

under different levels of irrigation and fertilization.

period (2014–2015), with a correlation coefficient R<sup>2</sup>

*DOI: http://dx.doi.org/10.5772/intechopen.93741*

The AquaCrop model is capable of simulating these parameters. Overall, the agreement between simulated and observed vegetation cover and biomass *Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen… DOI: http://dx.doi.org/10.5772/intechopen.93741*

**Figure 7.** *Variation in water productivity of lettuce under different levels of fertilization.*


**Table 3.**

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

*Effect of different levels of fertilization on yield for the two growing seasons.*

**3.6 Effect of fertilization on water productivity**

*Variation in nitrogen use efficiency by lettuce, expressed as a percentage (%).*

**3.7 Calibration of the AquaCrop model**

*3.7.1 Canopy cover and dry biomass*

Above this level, the increase in the nitrogen rate is not significant.

**Figure 7** shows the variation in water productivity (WP), soil evaporation (Es) and transpiration (Tr) of the lettuce crop under different levels of fertilization. This variation is supported by the analysis of variance, which showed a very highly significant effect (p < 0.001) of these parameters (WP, Es and Tr), in relation to the increase in the doses of nitrogen applied. The maximum values of WP and Tr are reached at the dose of 120 kg N/ha, for the two companions 2014–2015 (WP = 8.95 kg/m3; Tr =51.4 mm) and 2015–2016 (9.57 kg/m3; Tr = 55.80 mm).

Experimental results of yield, canopy cover and dry above-ground biomass under different levels of fertilization are presented in **Table 3**. The AquaCrop model (V. 6.1) was calibrated using the crop data set obtained from the T3 treatment (120 kg N/ha). The lowest dry yield and dry aboveground biomass observed were 4.021 t/ha and 4.125 t/ha under the T1 treatment (0 kg N/ha), and the highest were 8.854 t/ha and 9.320 t/ha under the T3 treatment (120 kg N/ha), respectively. The AquaCrop model is capable of simulating these parameters. Overall, the agreement between simulated and observed vegetation cover and biomass

**106**

**Figure 5.**

**Figure 6.**

*Biomass calibration results, yield, and maximum canopy cover under different levels of fertilization in 2015–2016.*

is satisfactory with 0.64 < R2 < 0.81, 18 < nRMSE > 46.3 and 0.78 < d < 0.94; 0.92 < R2 < 0.94, 21.6 < nRMSE <34.5, 0.91 < d < 0.96 (**Table 4**).

**Figure 8** shows the comparison between simulated and observed canopy cover (CC) and dry above-ground biomass (B) for the calibration period (2015–2016). This figure shows that there is a close correspondence between observed and simulated CC and B. It is also important to note that the AquaCrop model correctly simulates CC from seeding to the maximum growth phase at which CCx is reached. This observation has been reported in several studies [35–37]. From **Figure 8**, it is clear that both parameters (CC) and B were overestimated by the AquaCrop model. In a recent study [38], it was shown that the AquaCrop model overestimated the cabbage canopy under different irrigation regimes. Nikolaus [39] also noted a slight (10%) but systematic overestimation of the amount of rice biomass conducted under different levels of irrigation and fertilization.

#### *3.7.2 Yield*

Observed and simulated lettuce yields are shown in **Figure 9**. The observed yields for treatments T1, T2, T3 and T4 are, respectively, 4.214; 5.187; 6.942 and 6.214 t/ha, while the simulated yields are 4.897; 5.981; 7.414 and 6.987 for the trial period (2014–2015), with a correlation coefficient R<sup>2</sup> = 0.92. On the other hand, the yields observed and simulated under the four treatments for the trial period (2015–2016) are of the order of 4.021; 5.234; 7834 and 7.626 t/ha, while those simulated are of the order of 4.546; 5.785; 8.854 and 8.645, with a correlation coefficient R2 = 0.99. Analysis of statistical tests and linear regression indicated that the values


**Table 4.**

*Indicators of goodness of fit in estimating canopy cover and dry biomass.*

#### **Figure 8.**

*Canopy coverage (a) and dry biomass (b) simulated and measured for the calibration period (2015–2016) under different fertilization levels (T1, T2, T3, and T4).*

simulated by the AquaCrop model are in good agreement with those observed. Araya et al. [40] reported R<sup>2</sup> values >0.80 when simulating above-ground biomass and barley grain yield using AquaCrop.

**109**

*Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen…*

The management of nitrogen fertilization is a major issue for agricultural production while contributing to water and soil pollution. In this situation, the adoption of fertilization management strategies aimed at using efficient doses and increasing the effectiveness of their use becomes necessary. Crop models simulating yield under such conditions could be important tools for fertilizer management planning. To this end, the parameterization of the AquaCrop model to estimate the effect of fertility constraints on lettuce yield under different levels of fertilization was investigated. The model tended to overestimate canopy coverage for T3 (120 kg N/ha) and T4 (180 kg N/ha) treatments, but with reasonable statistical indices (nRMSE: 14.80 for T3 and 12.50 for T4). AquaCrop has confirmed that it is a very useful tool that can be used to optimize the N rates applied to the crops, to play on the management of the plot in order

*Simulated and observed lettuce yields under different levels of fertilization.*

*DOI: http://dx.doi.org/10.5772/intechopen.93741*

**4. Conclusion**

**Figure 9.**

to maximize yields.

*Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen… DOI: http://dx.doi.org/10.5772/intechopen.93741*

**Figure 9.** *Simulated and observed lettuce yields under different levels of fertilization.*

#### **4. Conclusion**

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

*Indicators of goodness of fit in estimating canopy cover and dry biomass.*

**Indicators CC (%) Dry biomass (t/ha)**

R2 0.81 0.71 0.66 0.64 0.92 0.98 0.94 0.94 NRMSE 18 35.5 41.4 46.3 34.5 21.6 25.6 25 EF 0.79 0.03 −0.13 −0.06 0.55 0.85 0.82 0.82 d 0.94 0.81 0.78 0.80 0.91 0.96 0.96 0.96

**T1 T2 T3 T4 T1 T2 T3 T4**

*Canopy coverage (a) and dry biomass (b) simulated and measured for the calibration period (2015–2016)* 

simulated by the AquaCrop model are in good agreement with those observed.

values >0.80 when simulating above-ground biomass

**108**

**Figure 8.**

**Table 4.**

*under different fertilization levels (T1, T2, T3, and T4).*

and barley grain yield using AquaCrop.

Araya et al. [40] reported R<sup>2</sup>

The management of nitrogen fertilization is a major issue for agricultural production while contributing to water and soil pollution. In this situation, the adoption of fertilization management strategies aimed at using efficient doses and increasing the effectiveness of their use becomes necessary. Crop models simulating yield under such conditions could be important tools for fertilizer management planning. To this end, the parameterization of the AquaCrop model to estimate the effect of fertility constraints on lettuce yield under different levels of fertilization was investigated. The model tended to overestimate canopy coverage for T3 (120 kg N/ha) and T4 (180 kg N/ha) treatments, but with reasonable statistical indices (nRMSE: 14.80 for T3 and 12.50 for T4). AquaCrop has confirmed that it is a very useful tool that can be used to optimize the N rates applied to the crops, to play on the management of the plot in order to maximize yields.

#### **Author details**

Mawhoub Amirouche1 \*, Dalila Smadhi<sup>2</sup> and Lakhdar Zella<sup>3</sup>

1 Department of Rural Engineering, Agricultural National High School, Algiers, Algeria

2 Division of Bioclimatology and Agricultural Hydraulic, National Institute for Agricultural Research, Algiers, Algeria

3 Department of Biotechnology, Faculty of Nature and life sciences, University of Saad Dahlab, Blida, Algeria

\*Address all correspondence to: mawhoub.amirouche@gmail.com

© 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.

**111**

*Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen…*

different growth stages using 15N isotope. Rice Science. 2015;**22**(5): 250-254. DOI: 10.1016/j.rsci.2015.09.005

[9] Mahfuzah NA, Khanif YM, Radziah O, Khairuddin AR. Timing of nitrogen uptake pattern by maize using 15N isotope technique at different growth stages. Bangladesh Journal of

Botany. 2017;**46**(1):329-334

fertilization of durum wheat

fields in the Doukkala region (Morocco). Proceedings of the Agronomic and Veterinary Institute.

1998;**18**(3):139-150

02008155/document

10.21273/HORTSCI.35.1.73

s10681-014- 1198-x

scitotenv.2017.04.062

[10] Bouaziz A, Soudi B. Nitrogenous

(*Triticum durum* Desf L.) in irrigated

[11] Yosser BZM. Analysis of the tradeoffs between production and water and nitrogen efficiency in Mediterranean cereal systems based on durum wheat [thesis]. France: Institut National d'Etudes Supérieures Agronomiques Montpellier SupAgro; 2017. Available from: https://tel.archives-ouvertes.fr/tel

[12] Charles AS. Response of lettuce to water and nitrogen on sand and the potential for leaching of nitrate-N. HortScience. 2000;**35**(1):73-77. DOI:

[13] Kerbiriou PJ, Stomph TJ, Lammerts van Bueren ET, Struik PC. Modelling concept of lettuce breeding for nutrient efficiency. Euphytica. 2004;**199**(1-2): 167-188. DOI: https://doi.org/10.1007/

[14] Pereira EIP, Conz R, Six J. Nitrogen utilization and environmental losses in organic greenhouse lettuce amended with two distinct biochars. The Science of the Total Environment. 2017;**598**:1169-1176. DOI: 10.1016/j.

[15] Chan-Navarrete R, Kawai A, Dolstra O, Lammerts van Bueren ET,

*DOI: http://dx.doi.org/10.5772/intechopen.93741*

[1] United Nations Organization (UNO). World Population Expected to Reach 9.6 Billion in 2050; 2013. p. 7. Available from: http://www.adequations.org/spip.

[2] Food and Agriculture Organization (FAO). Fertilizer Use by Crop in Algeria. 1st Ed. Rome, Italy: FAO; 2005. p. 56

[3] National Institute of Soil for Irrigation and Draining (INSID). Summary Note on the Actions Carried out by the National Institute of Soils, Irrigation and Drainage in the Context of Fertilization. 2009. p. 16. Available from: http://insid.dz/realisation/sol/

[4] Haffaf H, Benkherbache N,

[5] Saoudi M, Benkherbache N,

2016;**S1**:265-271

November 2004

Benniou R, Saoudi M. Study of nitrogen fertilization applied to the production of seeds of Triticum durum wheat (waha variety) in the semi-arid zone (M'sila). Agriculture Journal. 2016;**S1**:272-277

Benniou R, Haffaf H. Study of nitrogen fertilization applied to the production of *Hordeum vulgare* L barley seeds. (Tichedrett variety) in the semi-arid zone of M'sila. Agriculture Journal.

[6] Halilat MT. Effect of Potash and Nitrogen Fertilization on Wheat under Saharan Conditions. IPI regional workshop on Potassium and Fertigation

[7] Food and Agriculture Organization (FAO). Statistical Database. 2011. Available from: http://faostat.fao.org [Accessed: 01 December 2011]

development in West Asia and North Africa. Rabat, Morocco. 24-28

[8] Mohammad MH, Mohd KY, Radziah O, Samsuri AW.

Characterization of nitrogen uptake pattern in Malaysian rice MR219 at

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*Modeling of Nitrogen Use Efficiency in Lettuce Culture (*Lactuca sativa*): Isotopic Nitrogen… DOI: http://dx.doi.org/10.5772/intechopen.93741*

#### **References**

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

**110**

**Author details**

Algeria

Mawhoub Amirouche1

Agricultural Research, Algiers, Algeria

provided the original work is properly cited.

Saad Dahlab, Blida, Algeria

\*, Dalila Smadhi<sup>2</sup>

\*Address all correspondence to: mawhoub.amirouche@gmail.com

and Lakhdar Zella<sup>3</sup>

1 Department of Rural Engineering, Agricultural National High School, Algiers,

2 Division of Bioclimatology and Agricultural Hydraulic, National Institute for

3 Department of Biotechnology, Faculty of Nature and life sciences, University of

© 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,

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S0022-1694(02)00263-9

[25] Allen R, Pereira LS, Raes D, Smith M. Crop Evapotranspiration Guidelines for Computing Crop Water Requirements. Irrigation and Drainage Paper 56. Rome, Italy: FAO; 1998. p. 300

[26] Begoña B, Juan JR, Luis MC, Eva SR, María MR, Miguel AR, et al. Iodine application affects nitrogenuse efficiency of lettuce plants (*Lactuca sativa* L.). Acta Agriculturae Scandinavica Section B Soil and Plant Science. 2011;**61**(4):378-383. DOI: 10.1080/09064710.2010.492782

[27] Maurice EH, Robert FB, Darrel SM. Forages, 413-421. In: The Science of Grassland Agriculture. 4th Ed. Iowa, USA: Iowa State Univ. Press (Ames); 1985

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[29] Boroujerdnia M, Alemzadeh AN, Farideh SD. Effect of cultivars,

Islamic Azad University; 2005

Groeningen JW, Van Kessel C. Tracing 15N through landscapes: Potential uses and precautions. Journal of Hydrology. 2013;**272**:175-190. DOI: 10.1016/

Vienna, Austria; 2001

Van der Linden CG. Genetic diversity of nitrogen use efficiency inspinach (*Spinacia oleracea* L.) cultivars using the Ingestad model on hydroponics. Euphytica. 2014;**199**(1-2):155-166. DOI: 10.1007/s10681-014-1186-1

[16] Schulte EG, Dewi R, Nikus O, Horst WJ. Genotypic differences in nitrogen efficiency of white cabbage (*Brassica oleracea* L.). Plant and Soil. 2010;**328**:313-325. DOI: 10.1007/

[17] Saoud AA, Van CO, Hofman G. Uptake and balance of labelled fertilizer

[18] Collins HP, Delgado JA, Alva AK, Follett RF. Use of nitrogen-15 isotopic techniques to estimate nitrogen cycling from a mustard cover crop to potatoes. Agronomy Journal. 2007;**99**:27-35. DOI:

[19] Destain JP, Nathalie F, Dimitri X, Veronique R. The stable 15N isotope and the lysimeter, complementary tools for the study of nitrogen leaching in agricultural soils. Biotechnology, Agronomy, Society and Environment.

[20] Badr MA, Shedeed SI, Abou Hussein SD. Tomato fruit yield and nitrogen use efficiency as affected by drip irrigation method and rate of nitrogen in a hot dry climate. Journal of Applied Sciences Reseacch.

[21] DU Y-D, CAO H-X, LIU S-Q, GU X-B, CAO Y-X. Response of yield, quality, water and nitrogen use efficiency of tomato to different levels of water and nitrogen under drip irrigation in northwestern China. Journal of Integrative Agriculture. 2017;**16**(5):1153-1161. DOI: 10.1016/

nitrogen by potatoes. Fertilizer Research. 1992;**31**:351-353. DOI:

s11104-009-0111-1

10.1007/BF01051286

10.2134/agronj2005.0357

2010;**14**(S1):91-96

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**112**

[30] Amirouche M, Zella L, Smadhi D. Influence of nitrogen fertilization on lettuce yields (*Lactuca Sativa L*.) using the 15N isotope label. Agronomy Research. 2019;**17**(3):641-652. DOI: 10.15159/AR.19.118

[31] Tabatabaie SJ, Malakoutie MJ. Studies on the effect of the N, P and K-fertilizers on the potato yield and nitrate accumulation in potato tuber. Iran. Journal of Soil and Water Research. 1997;**11**:25-30

[32] Khelil MN, Rejeb S, Henchi B, Destain JP. Effect of fertilizer rate and water irrigation quality on the recovery of 15N–labeled fertilizer applied to Sudan grass. Agronomy for Sustainable Development. 2005;**25**:137-143. DOI: 10.1051/agro:2004065

[33] Kchaou R, Khelil MN, Gharbi F, Rejeb S, Rejeb MN, Henchi B, et al. Efficiency of use of nitrogen from residual sludge by sorghum feed. European Journal of Scientific Research. 2011;**54**:75-83

[34] Edith T, Lammerts VB, Paul CS. Diverse concepts of breeding for nitrogen use efficiency, a review. Agronomy for Sustainable Development. 2017;**37**:50. DOI: 10.1007/ s13593-017-0457-3

[35] Andarzian B, Bannayan M, Steduto P, Mazraeh H, Barati ME, Barati MA, et al. Validation and testing of the AquaCrop model under full and deficit irrigated wheat production in Iran. Agricultural Water Management. 2011;**100**:1-8. DOI: 10.1016/j.agwat.2011.08.023

[36] Xiangxiang W, Quanjiu W, Jun F, Qiuping F. Evaluation of the AquaCrop model for simulating the impact of water deficits and different irrigation regimes on the biomass and yield of winter wheat grown on China's loess plateau. Agricultural Water Management. 2013;**129**:95-104. DOI: 10.1016/j.agwat.2013.07.010

[37] Toumi J, Er-Raki S, Ezzahar J, Khabba S, Jarlan L, Chehbouni A. Performance assessment of AquaCrop model for estimating evapotranspiration, soil water content and grain yield of winter wheat in Tensift Al Haouz (Morocco): Application to irrigation management. Agricultural Water Management. 2016;**163**:219-235. DOI: 10.1016/j. agwat.2015.09.007

[38] Pawar GS, Kale MU, Lokhande JN. Response of AquaCrop model to different irrigation schedules for irrigated cabbage. Agricultural Research. 2017;**6**(1):73-81. DOI: 10.1007/s40003-016-0238-2

[39] Nikolaus B. Rice Production in Africa: Simulation of Rice Production and Water Productivity Using AquaCrop in an Inland Valley in Central Benin. Cotonou, Benin: Department of Hydrology and Water Resources Management, Christian-Albrechts-University Kiel, and Africa Rice Center; 2013

[40] Araya A, Habtub S, Hadguc KM, Kebedea A, Dejened T. Test of AquaCrop model in simulating biomass and yield of water deficient and irrigated barley (*Hordeum vulgare*). Agricultural Water Management. 2010;**97**:1838-1846. DOI: 10.1016/j.agwat.2010.06.021

**115**

**Chapter 8**

**Abstract**

for flower crop production.

nitrogen translocation, flowers

**1. Introduction**

Nitrogen in Flowers

*Soraya Ruamrungsri, Kanokwan Panjama,* 

This chapter explores the literature and research on nitrogen in flowers. An overview of nitrogen deficiency symptoms in some flowers, i.e., *Curcuma alismatifolia* (ornamental curcuma)*, Tagetes erecta* (marigold), *Zinnia violacea* (zinnia), and *Gomphrena globose* (gomphrena) were presented. Additionally, nitrogen uptake, translocation, and application in some flowers, i.e., ornamental curcuma*,* narcissus, orchids, and rose, were discussed in this chapter. Nitrogen affects the life cycle of flower, including vegetative and reproductive phases. Flower size, stem length, number of flowers per plant, and color were reduced by nitrogen deficiency. Therefore, the optimum level of nitrogen supply in each growth stage is important

Flower crops, similar to other horticultural crops, require optimum fertilizer for a good quality flower size, stem length, number of florets, stem, and petal color. Essential elements, especially nitrogen, play an important role in growth and development in each stage of the life cycle. Different genera have different nitrogen requirements. Generally, they have a nitrogen content that is enough for root emergence and shoot sprouting. However, flowers grown from seeds may require fertilizer as soon as root emergence. A lack of fertilizer supply will lead to severe nutrient deficiency. Seed germination starts with water uptake, and then food reserves, i.e., carbohydrates and storage proteins, are oxidized for the growth process [1]. Flower seedlings show deficiency symptoms when the fertilizer supply is not enough. Nitrogen is an especially important element in the life cycle of plants from seedlings to the vegetative stage and flowering until senescence. It affects flower qualities, such as size, stem length, and color. This chapter focuses on the role of nitrogen in some economic flower crops. Most information was derived from

**Keywords:** nitrogen deficiency, nitrogen uptake, nitrogen application,

our research experiments and some are unpublished data.

**2.1** *Curcuma alismatifolia* **(ornamental curcuma)**

**2. Nitrogen deficiency symptoms in different flower species**

Nitrogen deficiency (-N) affected the growth and characteristics of *C. alismatifolia* (**Table 1** and **Figure 1**). Most growth parameters, such as plant

*Takuji Ohyama and Chaiartid Inkham*

## **Chapter 8** Nitrogen in Flowers

*Soraya Ruamrungsri, Kanokwan Panjama, Takuji Ohyama and Chaiartid Inkham*

#### **Abstract**

This chapter explores the literature and research on nitrogen in flowers. An overview of nitrogen deficiency symptoms in some flowers, i.e., *Curcuma alismatifolia* (ornamental curcuma)*, Tagetes erecta* (marigold), *Zinnia violacea* (zinnia), and *Gomphrena globose* (gomphrena) were presented. Additionally, nitrogen uptake, translocation, and application in some flowers, i.e., ornamental curcuma*,* narcissus, orchids, and rose, were discussed in this chapter. Nitrogen affects the life cycle of flower, including vegetative and reproductive phases. Flower size, stem length, number of flowers per plant, and color were reduced by nitrogen deficiency. Therefore, the optimum level of nitrogen supply in each growth stage is important for flower crop production.

**Keywords:** nitrogen deficiency, nitrogen uptake, nitrogen application, nitrogen translocation, flowers

### **1. Introduction**

Flower crops, similar to other horticultural crops, require optimum fertilizer for a good quality flower size, stem length, number of florets, stem, and petal color. Essential elements, especially nitrogen, play an important role in growth and development in each stage of the life cycle. Different genera have different nitrogen requirements. Generally, they have a nitrogen content that is enough for root emergence and shoot sprouting. However, flowers grown from seeds may require fertilizer as soon as root emergence. A lack of fertilizer supply will lead to severe nutrient deficiency. Seed germination starts with water uptake, and then food reserves, i.e., carbohydrates and storage proteins, are oxidized for the growth process [1]. Flower seedlings show deficiency symptoms when the fertilizer supply is not enough. Nitrogen is an especially important element in the life cycle of plants from seedlings to the vegetative stage and flowering until senescence. It affects flower qualities, such as size, stem length, and color. This chapter focuses on the role of nitrogen in some economic flower crops. Most information was derived from our research experiments and some are unpublished data.

### **2. Nitrogen deficiency symptoms in different flower species**

#### **2.1** *Curcuma alismatifolia* **(ornamental curcuma)**

Nitrogen deficiency (-N) affected the growth and characteristics of *C. alismatifolia* (**Table 1** and **Figure 1**). Most growth parameters, such as plant


*\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05). NS = not significant.*

#### **Table 1.**

*Plant growth of* Curcuma alismatifolia *treated with complete nutrient solution or nitrogen deficiency (-N) at the flowering stage (12 weeks after planting).*

#### **Figure 1.**

*Growth and flower quality of* Curcuma alismatifolia *was affected by complete nutrient solution (control) and nitrogen deficiency (-N) treatment at the flowering stage (12 weeks after planting). (photo by Chaiartid Inkham).*

height, root length, leaf area, and total fresh and dry weight, were higher when plants were supplied with a complete nutrient solution, rather than -N treatment (**Table 1**). However, there was no significant difference in the number of leaves per plant between plants supplied with complete nutrient solution and -N treatment (**Table 1**). The characterization of nitrogen deficiency symptoms in *C. alismatiflolia* were evaluated at the flowering stage (12 weeks after planting). Leaves are the main plant part in which visual symptoms of the plant's response to nitrogen deficiency are usually observed. When there is a nitrogen deficit, older leaves of *C. alismatifolia* turn yellow and brown, while young leaves still appear green (**Figure 1**). The old leaves' green color intensity in -N treatment was lower than those treated with the complete nutrient solution (34.2 and 57.1 SPAD unit,

**117**

**Table 2.**

**Nutrient solution**

*NS = not significant.*

*Nitrogen in Flowers*

*DOI: http://dx.doi.org/10.5772/intechopen.98273*

and, consequently, the number of rhizomes [6].

ciency, thus increasing plant yield and flower quality [8].

**Inflorescence width (cm)**

Complete 70.7 b 6.9 a 11.8 a 34.3 a 1.0 a -N 78.3 a 5.8 a 9.9 b 29.4 a 1.0 a %CV 0.8 9.7 4.7 8.1 0 LSD 0.05 \* NS \* NS NS *\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05).* 

*Flower quality of* Curcuma alismatifolia *treated with complete nutrient solution or nitrogen deficiency (-N)* 

**Days to flowering (day)**

*at the flowering stage (12 weeks after planting).*

**2.2** *Tagetes erecta* **L. (Marigold)**

respectively). There was no significant difference among treatments in young leaves (**Table 1**). This result could explain nutrient remobilization processes in plants. Nitrogen is a macronutrient that is highly mobile in the phloem [2]; therefore, in N deficit conditions, nitrogen in old leaves of *C. alismatifolia* may be remobilized and translocate to young leaves. The remobilization of nutrients is frequently associated with foliar senescence, which makes nutrients available for

Nitrogen deficiency delays flowering in *C. alismatifolia* and decreased flower quality in term of inflorescent length. However, there were no significant differences in inflorescence width, stalk length, and number of inflorescences per plant (**Table 2** and **Figure 1**). Nitrogen deficiency delayed flowering in narrow-leafed lupin [4]. The production of *C. alismatifolia,* in terms of flower quality and rhizome yield, depends on the response to N fertilizers [5]. Nitrogen-deficient plants are stunted and the quality of their flowers and rhizomes is significantly decreased. The increase of nitrogen from 0 to 50 mg L−1 increased the number of flowering shoots

The overall growth parameters of marigold were decreased under nitrogen deficit conditions (**Table 1** and **Figure 2**). At 8 weeks after planting, plants in the -N treatment were stunted with a plant height of only 47.5 cm, which was 42.2 cm shorter than plants in the complete nutrient solution treatment. Moreover, there was a dramatic decrease in leaf area and the total fresh weight of marigolds grown under -N treatment when compared with complete nutrient solution treatment (decreasing 82 and 90%, respectively) (**Table 3**). Leaf green color intensity of marigold was detected both in young leaves and old leaves to evaluate visual symptoms of plants grown under -N conditions. The results showed that leaf green color intensity of marigold in both young and older leaves was lower when grown under -N treatment than grown under complete nutrient solution treatment (**Table 3**). In addition, the leaves of plants under -N treatment were smaller than those under complete nutrient solution treatment. Older leaves turned yellow, red and brown, while young leaves had symptoms of chlorosis and turned light yellow (**Figure 2**). Plant height, plant spread, and the number of primary branches per plant of African marigold increased significantly with the increase in nitrogen level from 0 to 30 g m−2 [7]. A suitable supply of N enhanced plant growth effi-

**Flower quality**

**Inflorescence length (cm)**

**Stalk length (cm)**

**No. inflorescence per plant**

younger plant organs and contributes to nutrient use efficiency [3].

#### *Nitrogen in Flowers DOI: http://dx.doi.org/10.5772/intechopen.98273*

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

**Root length (cm)**

**No. leaves per plant**

**Plant growth at flowering stage (12 weeks after planting)**

**Leaf green color intensity (SPAD unit)**

> **Young Leaf**

**leaf**

Complete 49.1 a 5.0 a 42.8 a 57.1 a 52.5 a 253.8 a 285.0 a 31.3 a -N 36.0 b 4.3 a 40.6 b 34.2 b 52.8 a 191.2 b 111.4 b 14.7 b %CV 8.7 8.8 0.9 10.8 4.9 10.6 11.0 12.4 LSD 0.05 \* NS \* \* NS \* \* \* *\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05).* 

*Plant growth of* Curcuma alismatifolia *treated with complete nutrient solution or nitrogen deficiency (-N) at* 

**Leaf area (cm2 )**

**(g) Old** 

**Total fresh weight (g)**

**Total dry weight** 

height, root length, leaf area, and total fresh and dry weight, were higher when plants were supplied with a complete nutrient solution, rather than -N treatment (**Table 1**). However, there was no significant difference in the number of leaves per plant between plants supplied with complete nutrient solution and -N treatment (**Table 1**). The characterization of nitrogen deficiency symptoms in *C. alismatiflolia* were evaluated at the flowering stage (12 weeks after planting). Leaves are the main plant part in which visual symptoms of the plant's response to nitrogen deficiency are usually observed. When there is a nitrogen deficit, older leaves of *C. alismatifolia* turn yellow and brown, while young leaves still appear green (**Figure 1**). The old leaves' green color intensity in -N treatment was lower than those treated with the complete nutrient solution (34.2 and 57.1 SPAD unit,

*Growth and flower quality of* Curcuma alismatifolia *was affected by complete nutrient solution (control) and nitrogen deficiency (-N) treatment at the flowering stage (12 weeks after planting). (photo by Chaiartid* 

**116**

**Figure 1.**

**Nutrient solution**

*NS = not significant.*

**Table 1.**

**Plant Height (cm)**

*the flowering stage (12 weeks after planting).*

*Inkham).*

respectively). There was no significant difference among treatments in young leaves (**Table 1**). This result could explain nutrient remobilization processes in plants. Nitrogen is a macronutrient that is highly mobile in the phloem [2]; therefore, in N deficit conditions, nitrogen in old leaves of *C. alismatifolia* may be remobilized and translocate to young leaves. The remobilization of nutrients is frequently associated with foliar senescence, which makes nutrients available for younger plant organs and contributes to nutrient use efficiency [3].

Nitrogen deficiency delays flowering in *C. alismatifolia* and decreased flower quality in term of inflorescent length. However, there were no significant differences in inflorescence width, stalk length, and number of inflorescences per plant (**Table 2** and **Figure 1**). Nitrogen deficiency delayed flowering in narrow-leafed lupin [4]. The production of *C. alismatifolia,* in terms of flower quality and rhizome yield, depends on the response to N fertilizers [5]. Nitrogen-deficient plants are stunted and the quality of their flowers and rhizomes is significantly decreased. The increase of nitrogen from 0 to 50 mg L−1 increased the number of flowering shoots and, consequently, the number of rhizomes [6].

#### **2.2** *Tagetes erecta* **L. (Marigold)**

The overall growth parameters of marigold were decreased under nitrogen deficit conditions (**Table 1** and **Figure 2**). At 8 weeks after planting, plants in the -N treatment were stunted with a plant height of only 47.5 cm, which was 42.2 cm shorter than plants in the complete nutrient solution treatment. Moreover, there was a dramatic decrease in leaf area and the total fresh weight of marigolds grown under -N treatment when compared with complete nutrient solution treatment (decreasing 82 and 90%, respectively) (**Table 3**). Leaf green color intensity of marigold was detected both in young leaves and old leaves to evaluate visual symptoms of plants grown under -N conditions. The results showed that leaf green color intensity of marigold in both young and older leaves was lower when grown under -N treatment than grown under complete nutrient solution treatment (**Table 3**). In addition, the leaves of plants under -N treatment were smaller than those under complete nutrient solution treatment. Older leaves turned yellow, red and brown, while young leaves had symptoms of chlorosis and turned light yellow (**Figure 2**). Plant height, plant spread, and the number of primary branches per plant of African marigold increased significantly with the increase in nitrogen level from 0 to 30 g m−2 [7]. A suitable supply of N enhanced plant growth efficiency, thus increasing plant yield and flower quality [8].


*\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05). NS = not significant.*

#### **Table 2.**

*Flower quality of* Curcuma alismatifolia *treated with complete nutrient solution or nitrogen deficiency (-N) at the flowering stage (12 weeks after planting).*

#### **Figure 2.**

*Growth and flower quality of marigold was affected by complete nutrient solution (control) and nitrogen deficiency (-N) treatments at the flowering stage (8 weeks after planting). (photo by Chaiartid Inkham).*


*\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05).*

#### **Table 3.**

*Plant growth of marigold treated with complete nutrient solution or nitrogen deficiency (-N) at the flowering stage (8 weeks after planting).*

Flowering of marigold was delayed under nitrogen deficiency condition (about 12 days delay when compared with complete nutrient treatment) (**Table 4**). Furthermore, the flower quality in terms of flower width, flower length, and stalk length were also reduced in plants in the -N treatment compared to those treated with the complete nutrient solution (**Table 4**, **Figure 2**). The flower yield of marigold was highly sensitive to nitrogen deficiency, since there was a 90% decrease in the number of flowers per plant when the plants were grown under -N treatment compared with the complete nutrient solution treatment (**Table 4**). In marigold (*Calendula officinalis* L. 'TOKAJ'), nitrogen fertilization had a significant impact on the number of flower heads per plant (especially on the second-rank branches) [9].

#### **2.3** *Zinnia violacea Cav.* **(zinnia)**

Nitrogen deficiency caused a decrease in plant height, number of leaves per plant, and root length of zinnia at 9 weeks after planting (**Table 5**). Additionally, yields of zinnia in terms of leaves area, total fresh weight, and total dry weight

**119**

increase) [10].

*Nitrogen in Flowers*

**Nutrient solution**

**Table 4.**

**Nutrient solution**

*NS = not significant.*

*stage (9 weeks after planting).*

**Table 5.**

*flowering stage (8 weeks after planting).*

**Plant Height (cm)**

**No. leaves per plant**

**Roots length (cm)**

*DOI: http://dx.doi.org/10.5772/intechopen.98273*

**Days to flowering (day)**

also decreased when grown plant under -N condition compared with those treated with complete nutrient solution; the percentage of reduction was 85, 81, and 84%, respectively (**Table 5**). Visual symptoms of nitrogen deficiency in zinnia were observed at the flowering stage. In -N treatment, plants were stunted with less branches (**Figure 3**). Old leaves in the -N treatment turned yellow, while young leaves remained green in both treatments (**Figure 3**). Visual symptoms observed on leaves had a similar trend with leaf green color intensity detected by a chlorophyll meter (SPAD) (**Table 5**). The results indicated that there was a significantly different leaf green color intensity in old leaves between plants treated with -N and complete nutrient solution. There was not a significant difference in young leaves (**Table 5**). Growth of *Zinnia elegans* Cv. Meteor increased with increasing nitrogen concentration from 0 to 20 g N/pot, i.e., plant growth rate (42% increase), plant height (28% increase), number of lateral shoots (56% increase), length of lateral shoots (17% increase), number of leaves (59% increase), and leaf area (40%

*Plant growth of zinnia treated with complete nutrient solution or nitrogen deficiency (-N) at the flowering* 

**Flower quality**

**Flower length (cm)**

**Stalk length (cm)**

**Leaf area (cm2 )**

**(g) Old** 

**Total fresh weight (g)**

**Total dry weight** 

**No. flowers per plant**

**Flower width (cm)**

Complete 55.7 b 8.5 a 4.8 a 12.7 a 10.0 a -N 67.7 a 4.8 b 2.0 b 4.5 b 1.0 b %CV 0.9 18.7 15.8 27.3 25.7 LSD 0.05 \* \* \* \* \* *\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05).*

*Flower quality of marigold treated with complete nutrient solution or nitrogen deficiency (-N) treatment at the* 

**leaf**

Complete 25.0 a 284.7 a 50.3 a 32.8 a 30.5 a 7,950.0 a 84.4 a 11.7 a -N 17.3 b 44.0 b 24.0 b 16.7 b 27.2 a 1,155.8 b 15.7 b 1.9 b %CV 10.7 17.4 15.7 14.3 9.4 9.6 9.9 19.4 LSD 0.05 \* \* \* \* NS \* \* \* *\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05).* 

**Plant growth at flowering stage (9 weeks after planting)**

**Young Leaf**

**Leaf green color intensity (SPAD unit)**

The flowering of zinnia was delayed for 15 days when plants were grown under -N treatment (**Table 6**). Nitrogen deficiency decreased flower quality in term of flower width (50% decrease), number of flower buds per plant (80% decrease),


#### **Table 4.**

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

Flowering of marigold was delayed under nitrogen deficiency condition (about

*Plant growth of marigold treated with complete nutrient solution or nitrogen deficiency (-N) at the flowering* 

Nitrogen deficiency caused a decrease in plant height, number of leaves per plant, and root length of zinnia at 9 weeks after planting (**Table 5**). Additionally, yields of zinnia in terms of leaves area, total fresh weight, and total dry weight

12 days delay when compared with complete nutrient treatment) (**Table 4**). Furthermore, the flower quality in terms of flower width, flower length, and stalk length were also reduced in plants in the -N treatment compared to those treated with the complete nutrient solution (**Table 4**, **Figure 2**). The flower yield of marigold was highly sensitive to nitrogen deficiency, since there was a 90% decrease in the number of flowers per plant when the plants were grown under -N treatment compared with the complete nutrient solution treatment (**Table 4**). In marigold (*Calendula officinalis* L. 'TOKAJ'), nitrogen fertilization had a significant impact on the number of flower heads per plant (especially on the second-rank branches) [9].

*Growth and flower quality of marigold was affected by complete nutrient solution (control) and nitrogen deficiency (-N) treatments at the flowering stage (8 weeks after planting). (photo by Chaiartid Inkham).*

> **Old leaf**

Complete 89.7 a 28.3 b 43.9 a 45.5 a 3,990.3 a 889.8 a -N 47.5 b 36.5 a 23.2 b 22.2 b 706.3 b 88.7 b %CV 8.9 5.9 14.1 19.0 40.3 2.7 LSD 0.05 \* \* \* \* \* \* *\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05).*

**Root length (cm)**

**Plant growth at flowering stage (8 weeks after planting)**

**Young Leaf**

**Leaf area (cm2 )**

**Total fresh weight (g)**

**Leaf green color intensity (SPAD unit)**

**118**

**Figure 2.**

**Table 3.**

*stage (8 weeks after planting).*

**Nutrient solution**

**Plant Height (cm)**

**2.3** *Zinnia violacea Cav.* **(zinnia)**

*Flower quality of marigold treated with complete nutrient solution or nitrogen deficiency (-N) treatment at the flowering stage (8 weeks after planting).*


*\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05). NS = not significant.*

#### **Table 5.**

*Plant growth of zinnia treated with complete nutrient solution or nitrogen deficiency (-N) at the flowering stage (9 weeks after planting).*

also decreased when grown plant under -N condition compared with those treated with complete nutrient solution; the percentage of reduction was 85, 81, and 84%, respectively (**Table 5**). Visual symptoms of nitrogen deficiency in zinnia were observed at the flowering stage. In -N treatment, plants were stunted with less branches (**Figure 3**). Old leaves in the -N treatment turned yellow, while young leaves remained green in both treatments (**Figure 3**). Visual symptoms observed on leaves had a similar trend with leaf green color intensity detected by a chlorophyll meter (SPAD) (**Table 5**). The results indicated that there was a significantly different leaf green color intensity in old leaves between plants treated with -N and complete nutrient solution. There was not a significant difference in young leaves (**Table 5**). Growth of *Zinnia elegans* Cv. Meteor increased with increasing nitrogen concentration from 0 to 20 g N/pot, i.e., plant growth rate (42% increase), plant height (28% increase), number of lateral shoots (56% increase), length of lateral shoots (17% increase), number of leaves (59% increase), and leaf area (40% increase) [10].

The flowering of zinnia was delayed for 15 days when plants were grown under -N treatment (**Table 6**). Nitrogen deficiency decreased flower quality in term of flower width (50% decrease), number of flower buds per plant (80% decrease),

#### **Figure 3.**

*Growth and flower quality of zinnia was affected by complete nutrient solution (control) and nitrogen deficiency (-N) treatment at the flowering stage (9 weeks after planting). (photo by Chaiartid Inkham).*


*\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05).*

#### **Table 6.**

*Flower quality of zinnia treated with complete nutrient solution or nitrogen deficiency (-N) at the flowering stage (9 weeks after planting).*

and number of flowers per plant (74% decrease) in zinnia (**Table 6**). Visual symptoms of zinnia flowers response to -N treatment were smaller flowers and a reduced number of petals compared to those treated with complete nutrient solution (**Table 6** and **Figure 3**). In *Zinnia elegans* cv. Meteor, flower quality increased when supplied with 10 g N/pot compared with 0 g N/pot (18% increase in number of flowers per plant, 52.5% increase in flower size, and blooming stage was prolonged for 11 days) [10]. A high dose of nitrogen (20 g N/pot) negatively impacted flowering when compared with those supplied 10 g N/pot, i.e., delay emergence of first flower for 8 days, 20% decrease in the number of flowers per plant, and 17.5% decrease in flower size [10].

**121**

**Figure 4.**

*Nitrogen in Flowers*

(**Figure 4**).

**Nutrient solution**

**Table 7.**

*DOI: http://dx.doi.org/10.5772/intechopen.98273*

The growth of gomphrena was affected by nitrogen deficiency (**Table 7**). In the -N treatment, plants were stunted, with plant height 34.4 cm lower than those in the complete nutrient treatment (49.3 cm). Shorter root length was also observed in plants under -N treatment. Leaf area, total fresh weight, and total dry weight were dramatically decreased when plants were grown under nitrogen deficiency conditions with 80, 85, and 85% reductions, respectively (**Table 7**). Visual symptoms of nitrogen deficiency were a changed leaf color. Both old and young leaves in the -N treatment turned yellow with SPAD values lower than those under complete nutrient treatment (**Table 7** and **Figure 4**). Moreover, a decrease in the number of new branches was observed when the plant was grown under -N treatment

**Plant growth at flowering stage (13 weeks after planting)**

**Leaf green color intensity (SPAD unit)**

> **Young Leaf**

**leaf**

Complete 49.3 a 49.3 a 59.7 a 30.6 a 37.7 a 1,251.8 a 250.6 a 39.2 a -N 34.4 b 21.7 b 32.6 b 22.1 b 26.6 b 254.2 b 38.0 b 6.0 b %CV 13.2 18.0 10.0 9.67 17.8 17.9 18.0 18.0 LSD 0.05 \* \* \* \* \* \* \* \* *\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05).*

*Plant growth of gomphrena treated with complete nutrient solution or nitrogen deficiency (-N) at the* 

*Growth and flower quality of Gomphrena was affected by complete nutrient solution (control) and nitrogen deficiency (-N) treatment at the flowering stage (13 weeks after planting). (photo by Chaiartid Inkham).*

**Leaf area (cm2 )**

**(g) Old** 

**Total fresh weight (g)**

**Total dry weight** 

**2.4** *Gomphrena globose* **(gomphrena)**

**Plant Height (cm)**

*flowering stage (13 weeks after planting).*

**No. leaves per plant**

**Root length (cm)**

#### **2.4** *Gomphrena globose* **(gomphrena)**

The growth of gomphrena was affected by nitrogen deficiency (**Table 7**). In the -N treatment, plants were stunted, with plant height 34.4 cm lower than those in the complete nutrient treatment (49.3 cm). Shorter root length was also observed in plants under -N treatment. Leaf area, total fresh weight, and total dry weight were dramatically decreased when plants were grown under nitrogen deficiency conditions with 80, 85, and 85% reductions, respectively (**Table 7**). Visual symptoms of nitrogen deficiency were a changed leaf color. Both old and young leaves in the -N treatment turned yellow with SPAD values lower than those under complete nutrient treatment (**Table 7** and **Figure 4**). Moreover, a decrease in the number of new branches was observed when the plant was grown under -N treatment (**Figure 4**).


*\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05).*

#### **Table 7.**

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

and number of flowers per plant (74% decrease) in zinnia (**Table 6**). Visual symptoms of zinnia flowers response to -N treatment were smaller flowers and a reduced number of petals compared to those treated with complete nutrient solution (**Table 6** and **Figure 3**). In *Zinnia elegans* cv. Meteor, flower quality increased when supplied with 10 g N/pot compared with 0 g N/pot (18% increase in number of flowers per plant, 52.5% increase in flower size, and blooming stage was prolonged for 11 days) [10]. A high dose of nitrogen (20 g N/pot) negatively impacted flowering when compared with those supplied 10 g N/pot, i.e., delay emergence of first flower for 8 days, 20% decrease in the number of flowers per plant, and 17.5%

*Flower quality of zinnia treated with complete nutrient solution or nitrogen deficiency (-N) at the flowering* 

*Growth and flower quality of zinnia was affected by complete nutrient solution (control) and nitrogen deficiency (-N) treatment at the flowering stage (9 weeks after planting). (photo by Chaiartid Inkham).*

**Days to flowering (day) Flowers width** 

**Flower quality**

**No. flower buds per plant**

**No. flowers per plant**

**(cm)**

Complete 54.3 b 6.2 a 54.0 a 26.3 a -N 69.3 a 3.1 b 10.7 b 6.7 b %CV 3.4 9.8 11.6 18.5 LSD 0.05 \* \* \* \* *\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05).*

**120**

**Figure 3.**

**Table 6.**

**Nutrient solution**

decrease in flower size [10].

*stage (9 weeks after planting).*

*Plant growth of gomphrena treated with complete nutrient solution or nitrogen deficiency (-N) at the flowering stage (13 weeks after planting).*

#### **Figure 4.**

*Growth and flower quality of Gomphrena was affected by complete nutrient solution (control) and nitrogen deficiency (-N) treatment at the flowering stage (13 weeks after planting). (photo by Chaiartid Inkham).*


#### **Table 8.**

*Flower quality of gomphrena treated with complete nutrient solution or nitrogen deficiency (-N) treatment at the flowering stage (13 weeks after planting).*

The flower quality of gomphrena was low under nitrogen deficiency conditions (**Table 8**). The flowering of gomphrena was delayed for 8 days when the plant lacked nitrogen. Additionally, flower size in terms of flower width, flower length, and stalk length were reduced under nitrogen deficit treatment. The number of flowers per plant also decreased by about 57% in the -N treatment compared with the complete nutrient treatment (**Table 8** and **Figure 4**).

#### **3. Uptake, translocation, and nitrogen application in different flower species**

Plants take up inorganic nitrogen, mostly in the form of ammonium (NH4 + ) and/or nitrate (NO3 − ). Uptake depends on the plant species and growth stage [11]. The translocation of N, including the free amino acid form, from roots to leaves could be done via the xylem. Some flowers can also utilize N via N2-fixation by endophytic bacteria, such as *Curcuma alismatifolia* and *Vanda*. Some studies have shown the uptake, translocation, and assimilation of N-forms in flowers at different growth stages using 15N-tracer feeding.

#### **3.1** *Curcuma alismatifolia*

*Curcuma alismatifolia*, commonly known as the Siam tulip, is a flower bulb in the family Zingiberaceae. It is an economical flower crop in Thailand. Growers export rhizomes and cut flowers to other countries, including Japan, the Netherlands, the USA. The inflorescence of this plant is showy with pink and greenish bracts on a long peduncle (**Figure 5A**). The storage organ is the underground part of the plant and is so-called rhizome modified from the stem and attached to some storage roots (**Figure 5B**).

The rhizome is a major organ to store N, while carbohydrates are mostly stored in storage roots. The N concentration in the stubbed rhizome is 41–45 mgN gDW−1, on average, while it was about 9–14 mgN gDW−1 in storage roots. Most of N in dormant rhizome was in PBS-insoluble form (79%), as a storage protein localized in the cytosol and cell wall, which presented as 10.6 and 12.0 kDa bands by SDS-PAGE staining. They contained five peptides and one peptide, respectively, when separated by 2D-PAGE. N in the storage roots was assimilated into different forms of free amino acids and protein compounds. The total free amino acid concentration in storage roots was higher than in the rhizome (343.8 and 109.0 μmol gDW−1, respectively)

**123**

*Nitrogen in Flowers*

**Figure 5.**

*DOI: http://dx.doi.org/10.5772/intechopen.98273*

when the plants were supplied with 50 mgN L−1. Most of the free amino acids in the rhizome and storage roots were arginine and glutamic acid, respectively. Proteins in the rhizome were at a higher concentration than those in storage roots with 197.4 and 46.7 mgN gDW−1, respectively. A lack of N reduced the protein and free amino acid concentration in both organs. The carbon content in *Curcuma* rhizomes and storage roots was about 0.9 and 2.48 g C plant−1, respectively, at the planting period, and it continuously decreased after planting. N and C stored in the rhizome was assimi-

*Inflorescence (A) and rhizome (B) of* Curcuma alismatifolia *Gagnep. (photo by Chaiartid Inkham).*

At *Curcuma* shoot sprouting, fertilizer was generally supplied, and N was translocated via roots and leaves. Anatomical study of *Curcuma* roots and leaves showed the different sizes of cortical cells and the vascular bundle, which were larger in roots than leaves. However, the number of vascular bundles in the roots were lower than in the leaves. The abaxial leaf surface presented less barriers than the adaxial surface [16]. The N-use efficiency of fertilizer (%) via roots was 51–57%, which was higher than that via leaves (7–10%). The research by 15N tracer revealed that N supply during the 1st and 2nd fully expanded leaf stage stimulated leaf growth, and N supply during the 3rd and 4th fully expanded leaf stage was translocate to utilize for flower blooming. N supply with 50 mg N L−1 increased the number of flowers and rhizomes compared with those at 25 mg N L−1 and lack of N supply. After translocation into plant organs, 81–97% of N was assimilated in an 80% ethanol insoluble fraction, mostly by proteins. N supply affected carbohydrate concentration in this plant, since nitrate reduction in roots requires carbohydrates for photosynthesis. The starch and sugar concentration in the rhizome and storage roots was high when *Curcuma* was grown under N deficiency. The translocation of carbohydrates to both storage organs was related to N deficiency conditions. From the vegetative stage until flowering, C content in leaves increased from 1.37 to 5.31 g C plant−1. The 13C exposure experiment revealed that C was accumulated in leaves during the vegetative stage and flowering, then it translocated to new storage organs (new rhizomes

lated and utilized for root emergence and shoot sprouting [12–15].

and storage roots) before plant dormancy [13, 15, 17, 18].

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

**Days to flowering (day)**

The flower quality of gomphrena was low under nitrogen deficiency conditions

*Flower quality of gomphrena treated with complete nutrient solution or nitrogen deficiency (-N) treatment at* 

**Flower quality**

**Flower length (cm)**

**Stalk length (cm)**

**No. flowers per plant**

(**Table 8**). The flowering of gomphrena was delayed for 8 days when the plant lacked nitrogen. Additionally, flower size in terms of flower width, flower length, and stalk length were reduced under nitrogen deficit treatment. The number of flowers per plant also decreased by about 57% in the -N treatment compared with

**Flower width (cm)**

complete 86.7 a 2.1 a 2.6 a 14.1 a 8.7 a -N 94.3 b 1.6 b 1.5 b 9.6 b 3.7 b %CV 0.6 6.9 16.1 8.1 29.6 LSD 0.05 \* \* \* \* \* *\*Means within the same column followed by different letters were significantly different in an LSD test; (p* ≤ *0.05).*

**3. Uptake, translocation, and nitrogen application in different flower** 

Plants take up inorganic nitrogen, mostly in the form of ammonium (NH4

The translocation of N, including the free amino acid form, from roots to leaves could be done via the xylem. Some flowers can also utilize N via N2-fixation by endophytic bacteria, such as *Curcuma alismatifolia* and *Vanda*. Some studies have shown the uptake, translocation, and assimilation of N-forms in flowers at different

*Curcuma alismatifolia*, commonly known as the Siam tulip, is a flower bulb in the family Zingiberaceae. It is an economical flower crop in Thailand. Growers export rhizomes and cut flowers to other countries, including Japan, the Netherlands, the USA. The inflorescence of this plant is showy with pink and greenish bracts on a long peduncle (**Figure 5A**). The storage organ is the underground part of the plant and is so-called rhizome modified from the stem and

The rhizome is a major organ to store N, while carbohydrates are mostly stored in storage roots. The N concentration in the stubbed rhizome is 41–45 mgN gDW−1, on average, while it was about 9–14 mgN gDW−1 in storage roots. Most of N in dormant rhizome was in PBS-insoluble form (79%), as a storage protein localized in the cytosol and cell wall, which presented as 10.6 and 12.0 kDa bands by SDS-PAGE staining. They contained five peptides and one peptide, respectively, when separated by 2D-PAGE. N in the storage roots was assimilated into different forms of free amino acids and protein compounds. The total free amino acid concentration in storage roots was higher than in the rhizome (343.8 and 109.0 μmol gDW−1, respectively)

). Uptake depends on the plant species and growth stage [11].

+ )

the complete nutrient treatment (**Table 8** and **Figure 4**).

**122**

**species**

**Nutrient solution**

**Table 8.**

and/or nitrate (NO3

**3.1** *Curcuma alismatifolia*

−

*the flowering stage (13 weeks after planting).*

growth stages using 15N-tracer feeding.

attached to some storage roots (**Figure 5B**).

**Figure 5.** *Inflorescence (A) and rhizome (B) of* Curcuma alismatifolia *Gagnep. (photo by Chaiartid Inkham).*

when the plants were supplied with 50 mgN L−1. Most of the free amino acids in the rhizome and storage roots were arginine and glutamic acid, respectively. Proteins in the rhizome were at a higher concentration than those in storage roots with 197.4 and 46.7 mgN gDW−1, respectively. A lack of N reduced the protein and free amino acid concentration in both organs. The carbon content in *Curcuma* rhizomes and storage roots was about 0.9 and 2.48 g C plant−1, respectively, at the planting period, and it continuously decreased after planting. N and C stored in the rhizome was assimilated and utilized for root emergence and shoot sprouting [12–15].

At *Curcuma* shoot sprouting, fertilizer was generally supplied, and N was translocated via roots and leaves. Anatomical study of *Curcuma* roots and leaves showed the different sizes of cortical cells and the vascular bundle, which were larger in roots than leaves. However, the number of vascular bundles in the roots were lower than in the leaves. The abaxial leaf surface presented less barriers than the adaxial surface [16]. The N-use efficiency of fertilizer (%) via roots was 51–57%, which was higher than that via leaves (7–10%). The research by 15N tracer revealed that N supply during the 1st and 2nd fully expanded leaf stage stimulated leaf growth, and N supply during the 3rd and 4th fully expanded leaf stage was translocate to utilize for flower blooming. N supply with 50 mg N L−1 increased the number of flowers and rhizomes compared with those at 25 mg N L−1 and lack of N supply. After translocation into plant organs, 81–97% of N was assimilated in an 80% ethanol insoluble fraction, mostly by proteins. N supply affected carbohydrate concentration in this plant, since nitrate reduction in roots requires carbohydrates for photosynthesis. The starch and sugar concentration in the rhizome and storage roots was high when *Curcuma* was grown under N deficiency. The translocation of carbohydrates to both storage organs was related to N deficiency conditions. From the vegetative stage until flowering, C content in leaves increased from 1.37 to 5.31 g C plant−1. The 13C exposure experiment revealed that C was accumulated in leaves during the vegetative stage and flowering, then it translocated to new storage organs (new rhizomes and storage roots) before plant dormancy [13, 15, 17, 18].

#### *3.1.1 Effect of temperature on N uptake*

Usually, N uptake and assimilation occur under normal temperature conditions. The optimum temperature for 15N uptake in *Curcuma alismatifolia* was 25–35°C [19]. Day and night temperature also affected N uptake and assimilation. The nitrate concentration in leaves, nitrate reductase activity in roots, and total free amino acid content was higher in a plants grown at 30/18°C (day/night temperature) than at 30/25°C. However, a low night temperature reduced the number of shoots per plant and inflorescence quality (spike length and stalk length) in this plant [20, 21].

#### *3.1.2 Response of curcuma to N application*

Plant dry matter contains 2–4% N. The most important inorganic forms of N are ammonium (NH4 + ) and nitrate (NO3 − ), which are converted to an organic form, such as proteins, amino acid, and nucleic acids. There are three steps of N turnover in plants: 1) the conversion of inorganic N to organic N; 2) synthesis of high molecular weight N, such as protein and nucleic acids; and 3) breakdown of nitrogenous macromolecules by hydrolyzing enzymes [1]. Therefore, N supply is essential for growth, flower quality, and yield. However, the response to N was dependent on plant species, soil condition, temperature, and nutrition level. A lack of N reduced growth and development of *Curcuma*. The number of flowers and rhizome yield also decreased under 0 mg N compared with 25 and 50 mg N [13]. A field experiment was carried out with different N application rates at 3.75, 7.5, 15, 30, and 60 g N/plant, and the results demonstrated that supra-optimal N application at 60 g N/plant reduced plant height, number of shoots/plant, leaf area, and plant dry weight, but leaf N and leaf chlorophyll content increased. The research revealed that the leaf critical N for *C. alismatifolia*, calculated by the Mitscherlichs model, was 1.51% [21] The optimum fertilizer rate was different depending on the growth stage. The optimum N rates were 234, 937, and 468 kg N/hectares at the vegetative stage (45–75 days after planting), flowering stage (105 days after planting), and before rhizome harvest (135–165 days after planting), respectively (**Figure 6**) [22].

#### *3.1.3 N2 fixation and IAA synthesis in curcuma by endophytic bacteria*

Nitrogen in the atmosphere that is fixed and converted to the organic form by microorganisms is termed N2 fixation. In *Curcuma*, N2 fixation by endophytic bacteria was first reported by Ruamrungsri et al. in 2009 [23]. The N-fixing rate varied depending on plant species. Eleven isolates were selected from *Curcuma alismatifolia* organs, such as the leaf, leaf base, and rhizome, and the N-fixing rate was 0.02–4.20 nmole C2H4/106 cells/hr. Seven isolates were derived from leaves, four isolates from the leaf base [23]. Isolates from the leaf base, i.e., ECS 202, identified using 16SrDNA, were *Sphingomonas pseudosanguinis* (99.2% similarity), ECS 203 was *Bacillus drentensis* and ECS 204 was *Bacillus methylotrophicus*. The colonization of these isolates was found in the intercellular spaces of different organs, i.e., roots, leaf base, and rhizome (**Figure 7**). Re-inoculation with these isolates into *Curcuma* plantlets derived from in vitro propagation was done by soaking roots in 106 cells/ml of these bacteria. Results showed that plant height, total leaf area (cm2 ), and N content in roots and leaves of plants inoculated with ECS 203 was higher than the control [24].

#### **3.2** *Narcissus*

*Narcissus* is a bulbs that has a storage organ that is modified from the leaf base in scales (**Figure 8**). Generally, the grower grows bulbs in autumn, and flowering

**125**

**Figure 6.**

*Nitrogen in Flowers*

*DOI: http://dx.doi.org/10.5772/intechopen.98273*

occurs in spring. Its roots are an unbranched system that emerges under low temperatures of 9°C. A lack of nitrogen decreased shoot height, root length, chlorophyll intensity, and dry weight of the plant. N-deficient leaves were yellow and small. N concentration was about 12.47 mg gDW−1, which was lower than the control plant (84.09 mgN gDW−1). Sugar content in N-deficient roots was also higher than the control (with N supply), indicating that N metabolism required carbohydrates as an energy source for nitrate reduction and assimilation. Nitrogen was absorbed from fertilizer application in the winter. The N absorbed by roots was translocated to other organs after shoot emergence to promote growth and development. Therefore, N supply was required after root emergence, although the N in the mother bulb was utilized for root growth and shoot sprouting. After shoot sprouting, leaves were the sink organ to derive N from the mother bulb and fertilizer until flower senescence. The uptake of ammonium and nitrate was studied in *Narcissus* using a 15N tracer. The

*Nitrogen application affected flower quality (A) and rhizome yield (B) of* Curcuma alismatifolia *(T1–T5* 

*were applied N at 234, 468, 937, 1,875, and 3,750 kg N/hectare). (Ruantip et al. [22]).*

results showed that at 2 days after fertilization with 1.0 mM of 15NH4

+

with 1.0 mM of 15NO3-N and 0.5 mM of 15NH4

'Garden Giant' roots could more rapidly uptake NH4

+

−

−



+




*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

) and nitrate (NO3

*3.1.3 N2 fixation and IAA synthesis in curcuma by endophytic bacteria*

vitro propagation was done by soaking roots in 106

plants inoculated with ECS 203 was higher than the control [24].

showed that plant height, total leaf area (cm2

Nitrogen in the atmosphere that is fixed and converted to the organic form by microorganisms is termed N2 fixation. In *Curcuma*, N2 fixation by endophytic bacteria was first reported by Ruamrungsri et al. in 2009 [23]. The N-fixing rate varied depending on plant species. Eleven isolates were selected from *Curcuma alismatifolia* organs, such as the leaf, leaf base, and rhizome, and the N-fixing rate was 0.02–4.20

the leaf base [23]. Isolates from the leaf base, i.e., ECS 202, identified using 16SrDNA, were *Sphingomonas pseudosanguinis* (99.2% similarity), ECS 203 was *Bacillus drentensis* and ECS 204 was *Bacillus methylotrophicus*. The colonization of these isolates was found in the intercellular spaces of different organs, i.e., roots, leaf base, and rhizome (**Figure 7**). Re-inoculation with these isolates into *Curcuma* plantlets derived from in

*Narcissus* is a bulbs that has a storage organ that is modified from the leaf base in scales (**Figure 8**). Generally, the grower grows bulbs in autumn, and flowering

cells/hr. Seven isolates were derived from leaves, four isolates from

cells/ml of these bacteria. Results

), and N content in roots and leaves of

Usually, N uptake and assimilation occur under normal temperature conditions. The optimum temperature for 15N uptake in *Curcuma alismatifolia* was 25–35°C [19]. Day and night temperature also affected N uptake and assimilation. The nitrate concentration in leaves, nitrate reductase activity in roots, and total free amino acid content was higher in a plants grown at 30/18°C (day/night temperature) than at 30/25°C. However, a low night temperature reduced the number of shoots per plant and inflorescence quality (spike length and stalk length) in this plant [20, 21].

Plant dry matter contains 2–4% N. The most important inorganic forms of N

), which are converted to an organic form,

−

such as proteins, amino acid, and nucleic acids. There are three steps of N turnover in plants: 1) the conversion of inorganic N to organic N; 2) synthesis of high molecular weight N, such as protein and nucleic acids; and 3) breakdown of nitrogenous macromolecules by hydrolyzing enzymes [1]. Therefore, N supply is essential for growth, flower quality, and yield. However, the response to N was dependent on plant species, soil condition, temperature, and nutrition level. A lack of N reduced growth and development of *Curcuma*. The number of flowers and rhizome yield also decreased under 0 mg N compared with 25 and 50 mg N [13]. A field experiment was carried out with different N application rates at 3.75, 7.5, 15, 30, and 60 g N/plant, and the results demonstrated that supra-optimal N application at 60 g N/plant reduced plant height, number of shoots/plant, leaf area, and plant dry weight, but leaf N and leaf chlorophyll content increased. The research revealed that the leaf critical N for *C. alismatifolia*, calculated by the Mitscherlichs model, was 1.51% [21] The optimum fertilizer rate was different depending on the growth stage. The optimum N rates were 234, 937, and 468 kg N/hectares at the vegetative stage (45–75 days after planting), flowering stage (105 days after planting), and before rhizome harvest (135–165 days after planting), respectively (**Figure 6**) [22].

*3.1.1 Effect of temperature on N uptake*

*3.1.2 Response of curcuma to N application*

+

are ammonium (NH4

**124**

nmole C2H4/106

**3.2** *Narcissus*

#### **Figure 6.**

*Nitrogen application affected flower quality (A) and rhizome yield (B) of* Curcuma alismatifolia *(T1–T5 were applied N at 234, 468, 937, 1,875, and 3,750 kg N/hectare). (Ruantip et al. [22]).*

occurs in spring. Its roots are an unbranched system that emerges under low temperatures of 9°C. A lack of nitrogen decreased shoot height, root length, chlorophyll intensity, and dry weight of the plant. N-deficient leaves were yellow and small. N concentration was about 12.47 mg gDW−1, which was lower than the control plant (84.09 mgN gDW−1). Sugar content in N-deficient roots was also higher than the control (with N supply), indicating that N metabolism required carbohydrates as an energy source for nitrate reduction and assimilation. Nitrogen was absorbed from fertilizer application in the winter. The N absorbed by roots was translocated to other organs after shoot emergence to promote growth and development. Therefore, N supply was required after root emergence, although the N in the mother bulb was utilized for root growth and shoot sprouting. After shoot sprouting, leaves were the sink organ to derive N from the mother bulb and fertilizer until flower senescence. The uptake of ammonium and nitrate was studied in *Narcissus* using a 15N tracer. The results showed that at 2 days after fertilization with 1.0 mM of 15NH4 + -N compared with 1.0 mM of 15NO3-N and 0.5 mM of 15NH4 + -N plus 0.5 mM of NO3 − -N, *Narcissus* 'Garden Giant' roots could more rapidly uptake NH4 + -N than NO3 − -N at 2 days after

#### **Figure 7.**

*Endophytic bacteria (A) isolate ECS 202 in the rhizome and (B) ECS 203 in the roots of* Curcuma alismatifolia*. (photo by Soraya Ruamrungsri).*

#### **Figure 8.**

*Tunicated bulb of narcissus with young flower inside (A) and unbranched roots (B). (photo by Soraya Ruamrungsri).*

fertilization, and the rate was equal between NH4 + -N and NO3 − -N for 4–7 days after fertilization. Moreover, the assimilation of N into free amino acids was also different based on the N-form. At 2 days after 15NH4 + -N fertilization, N in the roots was incorporated into free amino acids, mostly as glutamine, while asparagine and glutamine were the major assimilation forms of 15NO3 − -N supply. [25–28].

#### **3.3 Orchids**

The growth of orchids can be presented in two ways, sympodial and monopodial habits, depending on the genus. Some orchids have enlarged bulbous organs at the base of their leaves, called a pseudobulb, with a different shape. These organs store food and mineral nutrition for plant growth and development [29].

#### *3.3.1 Vanda*

*Vanda* is a tropical orchid comprised of 40 species. It is an economical orchid with a high export value. The roots of *Vanda* are aerial roots, freely hanging in the

**127**

*Nitrogen in Flowers*

prefers NO3

*3.3.2 Dendrobium*

*3.3.3 Phalaenopsis*

good quality flowers [33].

**Acknowledgements**

house and other facilities.

The authors declare no conflict of interest.

**Conflict of interest**

**3.4 Rose**

*DOI: http://dx.doi.org/10.5772/intechopen.98273*

plant nutrition in this plant is still rare.

occurred more rapidly than 15NH4


+

stems and roots at 30 days after fertilization [30].

nium and nitrate) to a sole N source [31].

development of this orchid. A combination of N sources (NO3

−

air. Growers use fertilizer applications to control flowering; however, research on

N source affected the uptake and translocation of N to various organs, and *Vanda*

Alanine distribution was high in leaves and roots at 7 days after feeding. Tyrosine distribution was predominant in leaves, while glutamine distribution was high in

In Thailand, *Dendrobium* is an important orchid genus for export. The main area for *Dendrobium* production is in the central region of Thailand, such as Ratchaburi, Nakhon, and Pathom provinces. Fertilizer application affected the growth and

moted the height of psuedobulbs, number of leaves and canes, spike length, and flowering percentage. The N concentration in leaves, roots, and psuedobulbs was 1.24, 0.97, and 0.61%, respectively. *Dendrobium* prefers a mixed N-source (ammo-

N level affected the growth of *Phalaenopsis* orchids. A concentration of 150–200 mg N L−1 increased the inflorescence length, stalk length, and number of flowers per stem. Supplying fertilizer with 21 N-21P2O5-21K2O at 150 mg L−1 once a week to young plants increased the leaf area, leaf dry weight, and N concentration [32]. However, when a high N concentration (200 mg N L−1) was supplied, the K concentration should also be supplied at 200 mg K L−1 to obtain healthy leaves and

Rose is the highest potential cut flower in the world market. Rose quality is graded

This work was partially supported by Chiang Mai University. Thanks to H.M. the King's Initiative Centre for flower and fruit propagation and the use of a green-

by stem length and flower size. N levels affected shoot height, shoot and flower diameter, flower fresh and dry weight, number of petals per flower, and flower yield. The N concentration in aboveground organs and roots was 19.4 and 20.9 mg N g DW−1 higher, respectively, when the plant was supplied with 200 mg N L−1. The lack of N decreased all quality parameters and showed deficiency symptoms. The optimum N concentration for roses was 200 mg N L−1 plant−1 once a week [34].

+

efficiency was higher in the mixed N source than the sole 15NO3 or 15NH4

The experiment on N uptake and assimilation in *Vanda* was carried out within 7 and 30 days after fertilization. The results revealed that plant uptake of 15NO3-N


−

and NH4

+ ) pro-


+ form. air. Growers use fertilizer applications to control flowering; however, research on plant nutrition in this plant is still rare.

The experiment on N uptake and assimilation in *Vanda* was carried out within 7 and 30 days after fertilization. The results revealed that plant uptake of 15NO3-N occurred more rapidly than 15NH4 + -N or their combination. However, 15N use efficiency was higher in the mixed N source than the sole 15NO3 or 15NH4 + form. N source affected the uptake and translocation of N to various organs, and *Vanda* prefers NO3 − -N to NH4 + -N. N assimilation in *Vanda* was different among organs. Alanine distribution was high in leaves and roots at 7 days after feeding. Tyrosine distribution was predominant in leaves, while glutamine distribution was high in stems and roots at 30 days after fertilization [30].

#### *3.3.2 Dendrobium*

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

*Endophytic bacteria (A) isolate ECS 202 in the rhizome and (B) ECS 203 in the roots of* Curcuma

fertilization, and the rate was equal between NH4

based on the N-form. At 2 days after 15NH4

were the major assimilation forms of 15NO3

+

fertilization. Moreover, the assimilation of N into free amino acids was also different

*Tunicated bulb of narcissus with young flower inside (A) and unbranched roots (B). (photo by Soraya* 

porated into free amino acids, mostly as glutamine, while asparagine and glutamine

+

−

The growth of orchids can be presented in two ways, sympodial and monopodial habits, depending on the genus. Some orchids have enlarged bulbous organs at the base of their leaves, called a pseudobulb, with a different shape. These organs store

*Vanda* is a tropical orchid comprised of 40 species. It is an economical orchid with a high export value. The roots of *Vanda* are aerial roots, freely hanging in the

food and mineral nutrition for plant growth and development [29].



−



**126**

**3.3 Orchids**

**Figure 8.**

*Ruamrungsri).*

**Figure 7.**

alismatifolia*. (photo by Soraya Ruamrungsri).*

*3.3.1 Vanda*

In Thailand, *Dendrobium* is an important orchid genus for export. The main area for *Dendrobium* production is in the central region of Thailand, such as Ratchaburi, Nakhon, and Pathom provinces. Fertilizer application affected the growth and development of this orchid. A combination of N sources (NO3 − and NH4 + ) promoted the height of psuedobulbs, number of leaves and canes, spike length, and flowering percentage. The N concentration in leaves, roots, and psuedobulbs was 1.24, 0.97, and 0.61%, respectively. *Dendrobium* prefers a mixed N-source (ammonium and nitrate) to a sole N source [31].

#### *3.3.3 Phalaenopsis*

N level affected the growth of *Phalaenopsis* orchids. A concentration of 150–200 mg N L−1 increased the inflorescence length, stalk length, and number of flowers per stem. Supplying fertilizer with 21 N-21P2O5-21K2O at 150 mg L−1 once a week to young plants increased the leaf area, leaf dry weight, and N concentration [32]. However, when a high N concentration (200 mg N L−1) was supplied, the K concentration should also be supplied at 200 mg K L−1 to obtain healthy leaves and good quality flowers [33].

#### **3.4 Rose**

Rose is the highest potential cut flower in the world market. Rose quality is graded by stem length and flower size. N levels affected shoot height, shoot and flower diameter, flower fresh and dry weight, number of petals per flower, and flower yield. The N concentration in aboveground organs and roots was 19.4 and 20.9 mg N g DW−1 higher, respectively, when the plant was supplied with 200 mg N L−1. The lack of N decreased all quality parameters and showed deficiency symptoms. The optimum N concentration for roses was 200 mg N L−1 plant−1 once a week [34].

#### **Acknowledgements**

This work was partially supported by Chiang Mai University. Thanks to H.M. the King's Initiative Centre for flower and fruit propagation and the use of a greenhouse and other facilities.

#### **Conflict of interest**

The authors declare no conflict of interest.

### **Author details**

Soraya Ruamrungsri1,2, Kanokwan Panjama1,2, Takuji Ohyama3 and Chaiartid Inkham2,4\*

1 Department of Plant and Soil Sciences, Faculty of Agriculture, Chiang Mai University, Chiang Mai, Thailand

2 Economic Flower Crop Research Cluster, Chiang Mai University, Chiang Mai, Thailand

3 Department of Agricultural Chemistry, Faculty of Applied Biosciences, Tokyo University of Agriculture, Japan

4 Science and Technology Research Institute, Chiang Mai University, Chiang Mai, Thailand

\*Address all correspondence to: sunwins111@hotmail.com

© 2021 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.

**129**

645-649.

*Nitrogen in Flowers*

**References**

*DOI: http://dx.doi.org/10.5772/intechopen.98273*

[1] Mengel K, Kirkby and EA. Principles of plant nutrition. International Potash

phosphorus levels on growth, yield and quality of African marigold. Annals of Plant Soil Research. 2012; 14(2): 153-155.

Agrobotanica. 2011; 64 (3): 29-34. DOI: https://doi.org/10.5586/aa.2011.027

[10] Khan MA, Ziaf K, Ahmad I. Influence of nitrogen on growth and flowering of *Zinnia elegans* Cv. Meteor. Asian Journal of Plant Sciences. 2004; 3 (5): 571-573. DOI: 10.3923/ajps.2004.571.573

[11] Marschner P. Marschner's mineral Nutrition of Higher Plants. Academic Press, Elsevier: Amsterdam; 2012. 651 p.

[12] Anuwong C, Ohtake N, Sueyoshi K,

[13] Ohtake N, Ruamrungsri S, Ito S, Sueyoshi K, Ohyama T, Apavatjrut P. Effect of nitrogen supply and

carbohydrate constituent accumulation in rhizomes and storage roots of *Curcuma alismatifolia* Gagnep. Soil Science and Plant Nutrition. 2006; 52: 711-716. https://doi.org/10.1111/ j.1747-0765.2006.00094.x

Sueyoshi K, Suwanthada C, Apavatjrut P, Ohyama T. Changes in nitrogenous compounds, carbohydrates and abscisic acid in *Curcuma alismatifolia* Gagnep. During dormancy. J of Horticultural Science & Biotechnology. 2001; 76 (1): 48-51. DOI: 10.1080/14620316.2001.

[14] Ruamrungsri S, Ohtake N,

[15] Khuankaew T, Ruamrungsri S, Ito S, Sato T, Ohtake N, Sueyoshi K,

11511325

Ruamrungsri S, Ohyama T. Characterisation of proteins in the storage organ of *Curcuma alismatifolia* Gagnep. Journal of horticultural Science & biotechnology. 2014; 89(5): 501-507. DOI: 10.1080/14620316.2014.11513112

[9] Król B. The effect of different nitrogen fertilization rates on yield and quality of marigold (*Calendula officinalis*

L. 'TOKAJ') raw material. Acta

[2] White P. Long-distance transport in the xylem and phloem. In: Marschner P. editors. Marschner's Mineral Nutrition of Higher Plants. 3rd ed. Berlin: Elsevier; 2012. p. 49-70. doi: 10.1016/ B978-0-12-384905-2.00003-0

[3] Avice JC, Etienne P. Leaf senescence and nitrogen remobilization efficiency in oilseed rape (*Brassica napus* L.). J. Exp. Bot. 2014; 65: 3813-3824. DOI:

[4] Ma Q, Longnecker N, Dracup M. Nitrogen deficiency slows leaf development and delays flowering in narrow-leafed lupin. Annals of Botany.

[5] Ruamrungsri S, Apavatjrut P. Effect of nutrient deficiency on growth and development of *Curcuma alismatifolia* Gagnep. In: Proceedings of the 3rd Symposium on the Family Zingiberaceae; 7-12 July 2003; Khon Kaen. Thailand;

[6] Ohtake N, Ruamrungsri S, Ito S, Sueyoshi K, Ohyama T Apavatjrut, P. Effect of nitrogen supply on nitrogen and carbohydrate constituent accumulation in rhizomes and storage roots of *Curcuma alismatifolia* Gagnep. Soil Sci. Plant Nutr.

2006; 52: 711-716. https://doi.org/ 10.1111/j.1747-0765.2006.00094.x

[7] Nain S, Beniwal BS, Dalal RPS, Sheoran S. Effect of nitrogen and phosphorus application on growth, flowering and yield of African marigold (*Tagetes erecta* L.) under semi-arid conditions of Haryana. Indian Journal of Ecology. 2016; 43 (Special Issue-2):

[8] Ahirwar MK, Ahirwar K, Shukla M. Effect of plant densities, nitrogen and

1997; 79: 403-409. https://doi. org/10.1006/anbo.1996.0361

Institute. Bern; 1987. 687 p.

10.1093/jxb/eru177

2003. p. 98-104.

## **References**

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

Soraya Ruamrungsri1,2, Kanokwan Panjama1,2, Takuji Ohyama3

\*Address all correspondence to: sunwins111@hotmail.com

1 Department of Plant and Soil Sciences, Faculty of Agriculture, Chiang Mai

2 Economic Flower Crop Research Cluster, Chiang Mai University, Chiang Mai,

3 Department of Agricultural Chemistry, Faculty of Applied Biosciences, Tokyo

4 Science and Technology Research Institute, Chiang Mai University, Chiang Mai,

© 2021 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,

**128**

**Author details**

Thailand

Thailand

and Chaiartid Inkham2,4\*

University, Chiang Mai, Thailand

University of Agriculture, Japan

provided the original work is properly cited.

[1] Mengel K, Kirkby and EA. Principles of plant nutrition. International Potash Institute. Bern; 1987. 687 p.

[2] White P. Long-distance transport in the xylem and phloem. In: Marschner P. editors. Marschner's Mineral Nutrition of Higher Plants. 3rd ed. Berlin: Elsevier; 2012. p. 49-70. doi: 10.1016/ B978-0-12-384905-2.00003-0

[3] Avice JC, Etienne P. Leaf senescence and nitrogen remobilization efficiency in oilseed rape (*Brassica napus* L.). J. Exp. Bot. 2014; 65: 3813-3824. DOI: 10.1093/jxb/eru177

[4] Ma Q, Longnecker N, Dracup M. Nitrogen deficiency slows leaf development and delays flowering in narrow-leafed lupin. Annals of Botany. 1997; 79: 403-409. https://doi. org/10.1006/anbo.1996.0361

[5] Ruamrungsri S, Apavatjrut P. Effect of nutrient deficiency on growth and development of *Curcuma alismatifolia* Gagnep. In: Proceedings of the 3rd Symposium on the Family Zingiberaceae; 7-12 July 2003; Khon Kaen. Thailand; 2003. p. 98-104.

[6] Ohtake N, Ruamrungsri S, Ito S, Sueyoshi K, Ohyama T Apavatjrut, P. Effect of nitrogen supply on nitrogen and carbohydrate constituent accumulation in rhizomes and storage roots of *Curcuma alismatifolia* Gagnep. Soil Sci. Plant Nutr. 2006; 52: 711-716. https://doi.org/ 10.1111/j.1747-0765.2006.00094.x

[7] Nain S, Beniwal BS, Dalal RPS, Sheoran S. Effect of nitrogen and phosphorus application on growth, flowering and yield of African marigold (*Tagetes erecta* L.) under semi-arid conditions of Haryana. Indian Journal of Ecology. 2016; 43 (Special Issue-2): 645-649.

[8] Ahirwar MK, Ahirwar K, Shukla M. Effect of plant densities, nitrogen and

phosphorus levels on growth, yield and quality of African marigold. Annals of Plant Soil Research. 2012; 14(2): 153-155.

[9] Król B. The effect of different nitrogen fertilization rates on yield and quality of marigold (*Calendula officinalis* L. 'TOKAJ') raw material. Acta Agrobotanica. 2011; 64 (3): 29-34. DOI: https://doi.org/10.5586/aa.2011.027

[10] Khan MA, Ziaf K, Ahmad I. Influence of nitrogen on growth and flowering of *Zinnia elegans* Cv. Meteor. Asian Journal of Plant Sciences. 2004; 3 (5): 571-573. DOI: 10.3923/ajps.2004.571.573

[11] Marschner P. Marschner's mineral Nutrition of Higher Plants. Academic Press, Elsevier: Amsterdam; 2012. 651 p.

[12] Anuwong C, Ohtake N, Sueyoshi K, Ruamrungsri S, Ohyama T. Characterisation of proteins in the storage organ of *Curcuma alismatifolia* Gagnep. Journal of horticultural Science & biotechnology. 2014; 89(5): 501-507. DOI: 10.1080/14620316.2014.11513112

[13] Ohtake N, Ruamrungsri S, Ito S, Sueyoshi K, Ohyama T, Apavatjrut P. Effect of nitrogen supply and carbohydrate constituent accumulation in rhizomes and storage roots of *Curcuma alismatifolia* Gagnep. Soil Science and Plant Nutrition. 2006; 52: 711-716. https://doi.org/10.1111/ j.1747-0765.2006.00094.x

[14] Ruamrungsri S, Ohtake N, Sueyoshi K, Suwanthada C, Apavatjrut P, Ohyama T. Changes in nitrogenous compounds, carbohydrates and abscisic acid in *Curcuma alismatifolia* Gagnep. During dormancy. J of Horticultural Science & Biotechnology. 2001; 76 (1): 48-51. DOI: 10.1080/14620316.2001. 11511325

[15] Khuankaew T, Ruamrungsri S, Ito S, Sato T, Ohtake N, Sueyoshi K, Ohyama T. Assimilation and translocation of nitrogen and carbon in *Curcuma alismatifolia* Gagnep. Plant Biol. 2009; 12: 414-423. DOI: 10.1111/ j.1438-8677.2009.00229.x

[16] Anuwong C, Ohyama T, Sueyoshi K, Ohtake N, Ruamrungsri S. Anatomical investigation of root and leaf of *Curcuma alismatifolia* Gagnep. Cv Chiang Mai Pink affecting N uptake. In: Proceedings of international graduate research conference Chiang Mai University. Chiang Mai, Thailand: 2014. p. 157-162.

[17] Marschner H. Mineral Nutrition of Higher Plants. Academic Press. Amsterdam; 1986. 674 p.

[18] Anuwong C, Ohyama T, Sueyoshi K, Ohtake N, Sato T, Ruamrungsri S. Uptake and translocation of nitrogen in Patumma (*Curcuma alismatifolia*) from foliar spray or root application. Journal of Plant Nutrition. 2017; 40(8): 1204- 1212. DOI: 10.1080/01904167.2016. 1264597

[19] Khuankaew T, Tanabata S, Yamamoto M, Ishikawa S, Tsutsumi K, Ohtake N, Sueyoshi K, Ohyama T, Sato T, Anuwong C, Ruamrungsri S. Temperature affects N and C assimilation and translocation in *Curcuma alismatifolia* Gagnep., The Journal of Horticultural Science and Biotechnology. 2014; 89(3): 287-292. DOI: 10.1080/14620316.2014.11513081

[20] Hongpakdee P, Ohtake N, Sueyoshi K, Ohyama T, S Ruamrungsri. Effects of low night temperature and short-day length on some phytohormones and nutrient status in *Curcuma alismatifolia* Gagnep. Thai journal of Agricultural Science. 2010; 43(3): 163-173.

[21] Inkham C, Sueyoshi K, Ohtake N, Ohyama T, Ruamrungsri S. Effect of temperature and nitrogen sources on growth and nitrogen assimilation of *Curcuma alismatifolia* Gagnep. Thai

journal of Agricultural Science. 2011; 44(3): 145-153.

[22] Ruantip T, Ruamrungsri S. Effects of nitrogen fertilizer levels on growth, flower and rhizome qualities of *Curcuma alismatifolia* Gagnep. In: Proceeding of the 2nd CMU graduate research conference. 26 November 2010. Graduate school. Chiang Mai University; 2010. p. 4.

[23] Ruamrungsri S, Hamtisong N, Choonluchanon S. Selection of endophytic bacteria from *Curcuma alismatifolia* for N2 fixation and IAA synthesis. In: Proceedings of the 3rd International meeting for the development of IPM in Asia and Africa, 7-9 December 2009; Bandar Lampung, Indonesia.

[24] Thepsukhon A, Choonluchanon S, Tajima S, Nomura M, Ruamrungsri S. Identification of endophytic bacteria associated with n2 fixation and indole acetic acid synthesis as growth promoters in *Curcuma alismatifolia* Gagnep. Journal of Plant Nutrition. 2013; 36(9): 1424-1438. DOI: 10.1080/01904167.2013.793712

[25] Ruamrungsri S, Ohyama T, Konno T, Ikarashi T. Deficiency of N, P, K, Ca, Mg or Fe mineral nutrients in *Narcissus* cv. 'Garden Giant'. Soil Science and Plant Nutrition. 1996; 42(4): 809-820. https:// doi.org/10.1080/00380768.1996.10416628

[26] Ruamrungsri S, Ohyama T, Ikarashi T. Nutrients, free amino acid and sugar contents in *Narcissus* roots affected by N, P, K deficiency during winter. Soil Science and Plant Nutrition. 1996; 42(4): 765-771.

[27] Ruamrungsri S, Ruamrungsri S, Ikarashi T, Ohyama T. Uptake, translocation and fractionation of nitrogen in *Narcissus* organs by using 15N. Acta Horticulturae. 1997; 430: 73-78. https://doi.org/10.1080/00380768 .1996.10416624

**131**

2010.878.41

*Nitrogen in Flowers*

0.11511227

1997. 370 p.

*DOI: http://dx.doi.org/10.5772/intechopen.98273*

[29] Hew CS, Yong JWH. The physiology of tropical orchids in relation to the industry. World Scientific: New Jersey;

[30] Panjama K, Ohyama T, Ohtake N, Sato T, Potapohn N, Sueyoshi K, Ruamrungsri S. Identifying N sources that affect N uptake and assimilation in *Vanda* hybrid using 15N tracers. Horticulture, Environment, and Biotechnology. 2018; 59(6): 805-813. DOI: 10.1007/s13580-018-0071-6

[31] Ruamrungsri S, Khuankaew T, T Ohyama. Nitrogen sources and its uptake in *Dendrobium* orchid by 15N Tracer study. Acta Horticulturae. 2014; 1025: 207-212. DOI: 10.17660/ActaHortic.2014.1025.30

[32] Ruamrungsri S, Samanit P,

Pornsawatchai T, Potapohn N, Fukai S. Effect of fertilizer application on nutritional concentrations and flower quality of Phalaenopsis hybrid. Acta Horticulturae. 2007; 755: 495-498. DOI: 10.17660/ActaHortic.2007.755.68

[33] Wang YT. Phalaenopsis mineral nutrition. Acta Horticulturae. 2010; 878: 321-334. DOI: 10.17660/ActaHortic.

[34] Tunprasas W, Ruamrungsri S. Effect of nitrogen on cut rose quality in potted production. J of Agriculture. Chiang Mai university. 2016; 32(3): 339-345.

[28] Ruamrungsri S, Ruamrungsri S, Ikarashi T, Ohyama T. Ammonium and nitrate assimilation in Narcissus roots. J. Hortic. Sci. Biotech. 2000; 75: 223-227. https://doi.org/10.1080/14620316.200

*Nitrogen in Flowers DOI: http://dx.doi.org/10.5772/intechopen.98273*

*Nitrogen in Agriculture - Physiological, Agricultural and Ecological Aspects*

journal of Agricultural Science. 2011;

[22] Ruantip T, Ruamrungsri S. Effects of nitrogen fertilizer levels on growth, flower and rhizome qualities of *Curcuma alismatifolia* Gagnep. In: Proceeding of the 2nd CMU graduate research

conference. 26 November 2010. Graduate school. Chiang Mai University; 2010. p. 4.

development of IPM in Asia and Africa, 7-9 December 2009; Bandar Lampung,

[24] Thepsukhon A, Choonluchanon S, Tajima S, Nomura M, Ruamrungsri S. Identification of endophytic bacteria associated with n2 fixation and indole acetic acid synthesis as growth promoters in *Curcuma alismatifolia* Gagnep. Journal of Plant Nutrition. 2013; 36(9): 1424-1438. DOI: 10.1080/01904167.2013.793712

[25] Ruamrungsri S, Ohyama T, Konno T, Ikarashi T. Deficiency of N, P, K, Ca, Mg or Fe mineral nutrients in *Narcissus* cv. 'Garden Giant'. Soil Science and Plant Nutrition. 1996; 42(4): 809-820. https:// doi.org/10.1080/00380768.1996.10416628

[26] Ruamrungsri S, Ohyama T, Ikarashi T. Nutrients, free amino acid and sugar contents in *Narcissus* roots affected by N, P, K deficiency during winter. Soil Science and Plant Nutrition.

[27] Ruamrungsri S, Ruamrungsri S, Ikarashi T, Ohyama T. Uptake, translocation and fractionation of nitrogen in *Narcissus* organs by using 15N. Acta Horticulturae. 1997; 430: 73-78. https://doi.org/10.1080/00380768

1996; 42(4): 765-771.

.1996.10416624

[23] Ruamrungsri S, Hamtisong N, Choonluchanon S. Selection of endophytic bacteria from *Curcuma alismatifolia* for N2 fixation and IAA synthesis. In: Proceedings of the 3rd International meeting for the

44(3): 145-153.

Indonesia.

Ohyama T. Assimilation and

j.1438-8677.2009.00229.x

translocation of nitrogen and carbon in *Curcuma alismatifolia* Gagnep. Plant Biol. 2009; 12: 414-423. DOI: 10.1111/

[16] Anuwong C, Ohyama T, Sueyoshi K, Ohtake N, Ruamrungsri S. Anatomical investigation of root and leaf of *Curcuma alismatifolia* Gagnep. Cv Chiang Mai Pink affecting N uptake. In: Proceedings of international graduate research conference Chiang Mai University. Chiang Mai, Thailand: 2014. p. 157-162.

[17] Marschner H. Mineral Nutrition of

[18] Anuwong C, Ohyama T, Sueyoshi K, Ohtake N, Sato T, Ruamrungsri S. Uptake and translocation of nitrogen in Patumma (*Curcuma alismatifolia*) from foliar spray or root application. Journal of Plant Nutrition. 2017; 40(8): 1204- 1212. DOI: 10.1080/01904167.2016.

Higher Plants. Academic Press. Amsterdam; 1986. 674 p.

[19] Khuankaew T, Tanabata S,

[20] Hongpakdee P, Ohtake N,

Sueyoshi K, Ohyama T, S Ruamrungsri. Effects of low night temperature and short-day length on some phytohormones and nutrient status in *Curcuma alismatifolia* Gagnep. Thai journal of Agricultural Science. 2010;

[21] Inkham C, Sueyoshi K, Ohtake N, Ohyama T, Ruamrungsri S. Effect of temperature and nitrogen sources on growth and nitrogen assimilation of *Curcuma alismatifolia* Gagnep. Thai

Yamamoto M, Ishikawa S, Tsutsumi K, Ohtake N, Sueyoshi K, Ohyama T, Sato T, Anuwong C, Ruamrungsri S. Temperature affects N and C assimilation and translocation in *Curcuma alismatifolia* Gagnep., The Journal of Horticultural Science and Biotechnology. 2014; 89(3): 287-292. DOI: 10.1080/14620316.2014.11513081

1264597

**130**

43(3): 163-173.

[28] Ruamrungsri S, Ruamrungsri S, Ikarashi T, Ohyama T. Ammonium and nitrate assimilation in Narcissus roots. J. Hortic. Sci. Biotech. 2000; 75: 223-227. https://doi.org/10.1080/14620316.200 0.11511227

[29] Hew CS, Yong JWH. The physiology of tropical orchids in relation to the industry. World Scientific: New Jersey; 1997. 370 p.

[30] Panjama K, Ohyama T, Ohtake N, Sato T, Potapohn N, Sueyoshi K, Ruamrungsri S. Identifying N sources that affect N uptake and assimilation in *Vanda* hybrid using 15N tracers. Horticulture, Environment, and Biotechnology. 2018; 59(6): 805-813. DOI: 10.1007/s13580-018-0071-6

[31] Ruamrungsri S, Khuankaew T, T Ohyama. Nitrogen sources and its uptake in *Dendrobium* orchid by 15N Tracer study. Acta Horticulturae. 2014; 1025: 207-212. DOI: 10.17660/ActaHortic.2014.1025.30

[32] Ruamrungsri S, Samanit P, Pornsawatchai T, Potapohn N, Fukai S. Effect of fertilizer application on nutritional concentrations and flower quality of Phalaenopsis hybrid. Acta Horticulturae. 2007; 755: 495-498. DOI: 10.17660/ActaHortic.2007.755.68

[33] Wang YT. Phalaenopsis mineral nutrition. Acta Horticulturae. 2010; 878: 321-334. DOI: 10.17660/ActaHortic. 2010.878.41

[34] Tunprasas W, Ruamrungsri S. Effect of nitrogen on cut rose quality in potted production. J of Agriculture. Chiang Mai university. 2016; 32(3): 339-345.

**133**

nutrition.

**Chapter 9**

**Abstract**

Nitrogen Storage in Crops: Case

Crop grains accumulate significant amounts of nitrogen in the form of storage proteins. Grain storage proteins are not only important in the aspects of germination but also, storage proteins are a valuable food source in human and animal nutrition. This chapter will give insight into genotype and growing conditions influencing the quantity and quality of storage proteins, primarily maize storage proteins the leading cereal by world production. Main storage proteins in cereals are prolamins, and in maize prolamins are called zeins located within the endosperm in protein agglomerations called protein bodies. Four main classes of zein proteins are: alpha, beta, gamma and delta zein. Each of four zein classes has a distinctive position and role within protein bodies. Prolamin proteins define nutritional value of maize grain not only via amino acid quality but also via starch availability. Starch, the most important energy component of maize grain, is located within starchprotein matrix. Within this matrix, starch granules are surrounded by protein bodies that limit starch availability. In this chapter, we will describe how zein proteins

*Marija Duvnjak, Kristina Kljak and Darko Grbeša*

influence characteristics of maize grain and nutritional value of maize.

maize nutritional value, animal nutrition

ization (0.05 Mg ha−1 year−1) [1].

**1. Introduction**

**Keywords:** maize grain, zein proteins, starch, amino acid quality, starch digestibility,

The protein content in cereal grain can vary greatly, from less than 6% to more than 20% in dry matter (DM), and this content depends on several factors such as the type of cereal, variety, agrotechnical conditions and others. These factors can be divided into two major groups, genotype and environment. Today, producers manipulate these factors to obtain grain of good quality and high protein content. For example, it has been found that the rate of maize yield gain is significantly higher after application of 220 kg N ha−1 (0.12 Mg ha−1 year−1) than without fertil-

In terms of their functions, there are three types of cereal grain proteins: structural proteins (as membrane proteins), metabolic proteins (as enzymes and enzyme inhibitors) and storage proteins, the largest fraction occurring primarily in starchy endosperm. Storage proteins account for 70–80% of the total protein content in grain and have a unique structure. The primary function of storage proteins is to supply grain embryo with nitrogen and amino acids during germination. However, these proteins are also a valuable food and feed source in human and animal

Study of Zeins in Maize

#### **Chapter 9**
