Nitrogen Fertilization in Blackberry

*Ivan dos Santos Pereira, Adilson Luis Bamberg, Carlos Augusto Posser Silveira, Luis Eduardo Corrêa Antunes and Rogério Oliveira de Sousa*

#### **Abstract**

Nutrition studies for blackberry crop are scarce worldwide. This chapter presents several aspects of nitrogen (N) in blackberry (*Rubus* spp.) nutrition. Soil characteristics that can influence nitrogen fertilization are the large discrepancies in the rates recommended in the literature, forms and times of application, sources of nitrogen, differences between cultivars and the main symptoms of N deficiency. The impact of moderate and severe nitrogen deficiency on vegetative growth and yield of 'Tupy' blackberry is also presented. In addition, a nitrogen fertilization recommendation system is proposed, based on the organic matter content of the soil, the age of the plants, and the expected productivity of the cultivars.

**Keywords:** *Rubus* spp., nutritional requirements, cultivar differences, nutritional deficiency, soil organic matter, fertilization recommendation

#### **1. Introduction**

Nitrogen (N) is the mineral element that plants generally need in greater quantity, since they serve to form components of plant cells, such as amino acids and nucleic acids, besides participating in the chlorophyll molecule [1]. N deficiency rapidly reduces plant growth, as it causes reduction of cell division and expansion, leaf area, and photosynthesis [2].

In blackberry (*Rubus* spp.), N is the most abundant element and plays a major role in its growth, development, and productivity [3–6]. The optimum leaf content required for a satisfactory performance of blackberry varies from 2.2 to 3.0% of the dry matter of the leaves [7, 8].

The need for N supply may vary according to soil organic matter (SOM) content, yield, growth habit, age, and cultivar [8, 9]. The N rates recommended in the literature vary widely, mainly due to differences between cultivars and soil characteristics, but another important factor is the age of the plants. In the first years, the productive capacity of the plants is smaller, and therefore the demand for nitrogen is also lower. High N rates in the first 2 years can reduce fruit quality and increase disease incidence. On the other hand, low rates from the third year make it difficult to obtain high yield.

Nitrogen fertilization provides immediate effect (same season) and residual (next season). The immediate effect is mainly on the productive capacity of the floricans and the size of the fruits. Already in the following season, the greatest influence is on the growth and formation of primocanes and floral buds [10].

Considering the N importance to blackberry, this chapter aims to review the dynamics of nitrogen fertilization and present a recommendation proposal.

#### **2. Soil characteristics**

The recommended soil pH for blackberries is 5.5 to 6.5 [7, 8, 11]. The availability of N from fertilizers mainly depends on the fertilizer source [12] and the method of application [13]. Soil pH affects nitrification, with ammonium-N sources converted more rapidly to nitrate-N at a pH of 6.0 than at 5.5 [8].

Another issue that must be observed in relation to pH is the presence of aluminum (Al3+) in the soil. In these cases it is essential that the pH is raised to values close to 6.0 through liming, avoiding problems with phytotoxicity by aluminum.

The ideal soil organic matter content is around 2.0 to 4.5% [3, 14, 6]. However, what is observed is that the blackberry is a rustic species, which can be cultivated in soils with a wide range of SOM levels. The difference is that in soils with low levels, less than 2%, there is a need for special care in relation to nitrogen fertilization. On the other hand, when the content of SOM is high, more than 4.5%, the vigor of the plants is driven in a way that increases the frequency and intensity of pruning, as well as decreases the need for N fertilization.

Some studies indicate a lack of effect of fertilization on yield in soils with a SOM content of 3.9%, while in soil with a SOM content of 1.1%, there was a linear increase of the yield in response to increasing rates of N [4, 15]. These results suggest an important influence of SOM on the response of blackberry to fertilization with N.

#### **3. Nitrogen fertilizer rate**

There are significant variations in the nitrogen rates recommended by the literature, with values ranging between 0.0 and 200 kg ha−1 . Such differences may be related to factors such as soil characteristics, climate, and genotypes [3, 5].

Some authors recommend the application of 34–56 kg ha−1 in the first year, regardless of growth habit, and in the following years, the dose would be from 56 to 78 kg ha−1 for training and semi-erect cultivars and from 56 to 90 kg ha−1 for cultivars of erect habit [8].

Other indications for semi-erect and erect cultivars are 25–45 kg ha−1 at the establishment and 45–70 kg ha−1 in subsequent years [16, 17]. The recommendations for training cultivars are between 25 and 45 kg ha−1 in the year of establishment and between 45 and 60 kg ha−1 in the following seasons [17].

Considering that Tupy and Xavante has semi-erect and erect growth habit, respectively, there was an inversion of the logic indicated in the recommendations of the literature. In this case, probably the greatest nutritional need of the cultivar Tupy may come as a result of its greater yield and export of nutrients, about 40% higher than that obtained with Xavante.

Another feature that diverges between these two genotypes is the presence of thorns. There is a need for new studies in order to verify if there is a possible relationship of this characteristic with the nutritional need.

The amount of nutrients applied in blackberry cultivation can also vary with the age of the plants. The application of N in the year of establishment of the crop is controversial; there are authors who do not recommend the application, due to the risk of damages to the vegetative buds and others that, although emphasizing

**59**

**Figure 1.**

*productivity (c) [9].*

*Nitrogen Fertilization in Blackberry*

application of up to 56 kg ha−1

efficiency rate was 109 kg ha−1

material removed) [7, 14].

Al3+ content was higher than 0.66 cmolc dm−3

the pH and Al3+ content of the soil should be monitored.

responsible for fruit production in the following year.

**4. Nitrogen fertilizer application**

155 kg ha−1

*DOI: http://dx.doi.org/10.5772/intechopen.82409*

that the application is less necessary than in the subsequent years, suggest the

differential fertilization strategy, depending on the age of the plants.

justified by the high demand that the plant has in relation to this nutrient.

the south of Brazil, in an area with a 1.1% SOM content, the maximum productive

The N application is usually done in granular form while planting directly in the row but can be carried out during crop development, via fertigation or foliar fertilization [14, 17]. In a study comparing the application methods, it was concluded that the granular fertilization in the spring had a better result than the drip fertigation, mainly due to the higher leaching observed in the drip method [13]. Although it can be used, foliar fertilization has not presented satisfactory results [8], which is

About the N sources, blackberry responds satisfactorily to several, but ammonium nitrate, urea, and ammonium sulfate are the most commonly used sources [3, 17]. In Brazil, the ammonium sulfate is currently recommended, as blackberries also demand important sulfur amount (about 3 kg per ton of fruit and pruning

However, research results indicate that after 3 years of using ammonium sulfate as N source, there is a significant reduction of pH (**Figure 1a**) and increase of Al3+ content (**Figure 1b**) in the soil, which may have a negative impact on the productivity (**Figure 1c**). In this specific case, when the soil pH was lower than 4.7 and the

However, further studies on different soil types should be carried out in order to confirm this relationship. Therefore, when using ammonium sulfate as N source,

As for the time of application, N should be applied in the spring and after harvest [7, 14, 18]. Fertilization carried out at the end of winter or early spring aims to provide fruit production and the growth of primocanes, new stems that will be

N has an important role in the formation of "primocane" numbers since it stimulates budding of crown buds, thus impacting the number of stems and the yield of the next season [3]. Maintaining the proper number of stems over the years is important. There is a positive correlation between the number of stems and the

*Influence of ammonium sulfate rates on pH (a), aluminum (Al3+) concentration (b), and blackberry 'Tupy'*

at the third crop [4]. Thus, these results indicate that there is a need for

[8, 14]. On the other hand, in a study carried out in

in the first harvest after planting and increased up to

, blackberry production decreased.

*Nitrogen Fixation*

**2. Soil characteristics**

floricans and the size of the fruits. Already in the following season, the greatest influence is on the growth and formation of primocanes and floral buds [10]. Considering the N importance to blackberry, this chapter aims to review the

The recommended soil pH for blackberries is 5.5 to 6.5 [7, 8, 11]. The availability of N from fertilizers mainly depends on the fertilizer source [12] and the method of application [13]. Soil pH affects nitrification, with ammonium-N sources converted

Another issue that must be observed in relation to pH is the presence of aluminum (Al3+) in the soil. In these cases it is essential that the pH is raised to values close to 6.0 through liming, avoiding problems with phytotoxicity by aluminum. The ideal soil organic matter content is around 2.0 to 4.5% [3, 14, 6]. However, what is observed is that the blackberry is a rustic species, which can be cultivated in soils with a wide range of SOM levels. The difference is that in soils with low levels, less than 2%, there is a need for special care in relation to nitrogen fertilization. On the other hand, when the content of SOM is high, more than 4.5%, the vigor of the plants is driven in a way that increases the frequency and intensity of pruning, as

Some studies indicate a lack of effect of fertilization on yield in soils with a SOM content of 3.9%, while in soil with a SOM content of 1.1%, there was a linear increase of the yield in response to increasing rates of N [4, 15]. These results suggest an important influence of SOM on the response of blackberry to fertilization with N.

There are significant variations in the nitrogen rates recommended by the

regardless of growth habit, and in the following years, the dose would be from 56

Considering that Tupy and Xavante has semi-erect and erect growth habit, respectively, there was an inversion of the logic indicated in the recommendations of the literature. In this case, probably the greatest nutritional need of the cultivar Tupy may come as a result of its greater yield and export of nutrients, about 40%

Another feature that diverges between these two genotypes is the presence of thorns. There is a need for new studies in order to verify if there is a possible

The amount of nutrients applied in blackberry cultivation can also vary with the age of the plants. The application of N in the year of establishment of the crop is controversial; there are authors who do not recommend the application, due to the risk of damages to the vegetative buds and others that, although emphasizing

for training and semi-erect cultivars and from 56 to 90 kg ha−1

in subsequent years [16, 17]. The recommenda-

in the following seasons [17].

be related to factors such as soil characteristics, climate, and genotypes [3, 5].

Other indications for semi-erect and erect cultivars are 25–45 kg ha−1

. Such differences may

in the first year,

in the year of establish-

for

at the

literature, with values ranging between 0.0 and 200 kg ha−1

tions for training cultivars are between 25 and 45 kg ha−1

relationship of this characteristic with the nutritional need.

Some authors recommend the application of 34–56 kg ha−1

dynamics of nitrogen fertilization and present a recommendation proposal.

more rapidly to nitrate-N at a pH of 6.0 than at 5.5 [8].

well as decreases the need for N fertilization.

**3. Nitrogen fertilizer rate**

cultivars of erect habit [8].

establishment and 45–70 kg ha−1

ment and between 45 and 60 kg ha−1

higher than that obtained with Xavante.

to 78 kg ha−1

**58**

that the application is less necessary than in the subsequent years, suggest the application of up to 56 kg ha−1 [8, 14]. On the other hand, in a study carried out in the south of Brazil, in an area with a 1.1% SOM content, the maximum productive efficiency rate was 109 kg ha−1 in the first harvest after planting and increased up to 155 kg ha−1 at the third crop [4]. Thus, these results indicate that there is a need for differential fertilization strategy, depending on the age of the plants.

#### **4. Nitrogen fertilizer application**

The N application is usually done in granular form while planting directly in the row but can be carried out during crop development, via fertigation or foliar fertilization [14, 17]. In a study comparing the application methods, it was concluded that the granular fertilization in the spring had a better result than the drip fertigation, mainly due to the higher leaching observed in the drip method [13]. Although it can be used, foliar fertilization has not presented satisfactory results [8], which is justified by the high demand that the plant has in relation to this nutrient.

About the N sources, blackberry responds satisfactorily to several, but ammonium nitrate, urea, and ammonium sulfate are the most commonly used sources [3, 17]. In Brazil, the ammonium sulfate is currently recommended, as blackberries also demand important sulfur amount (about 3 kg per ton of fruit and pruning material removed) [7, 14].

However, research results indicate that after 3 years of using ammonium sulfate as N source, there is a significant reduction of pH (**Figure 1a**) and increase of Al3+ content (**Figure 1b**) in the soil, which may have a negative impact on the productivity (**Figure 1c**). In this specific case, when the soil pH was lower than 4.7 and the Al3+ content was higher than 0.66 cmolc dm−3 , blackberry production decreased. However, further studies on different soil types should be carried out in order to confirm this relationship. Therefore, when using ammonium sulfate as N source, the pH and Al3+ content of the soil should be monitored.

As for the time of application, N should be applied in the spring and after harvest [7, 14, 18]. Fertilization carried out at the end of winter or early spring aims to provide fruit production and the growth of primocanes, new stems that will be responsible for fruit production in the following year.

N has an important role in the formation of "primocane" numbers since it stimulates budding of crown buds, thus impacting the number of stems and the yield of the next season [3]. Maintaining the proper number of stems over the years is important. There is a positive correlation between the number of stems and the

#### **Figure 1.**

*Influence of ammonium sulfate rates on pH (a), aluminum (Al3+) concentration (b), and blackberry 'Tupy' productivity (c) [9].*

yield of the blackberry, that is, the larger the number of stems (up to 12 stems m−1 ), the greater the productivity [19].

On the other hand, postharvest fertilization, usually performed after postharvest pruning, has the function of stimulating the development of primocanes, inducing the formation of vigorous stems and thus capable of supporting high yields and larger fruits in the next season. In blackberry cultivation, there is a significant correlation between stem diameter and fruit size, and more vigorous stems have potential for larger fruit production [19, 20].

#### **5. Nutritional differences between cultivars**

It has been observed in the literature that blackberry cultivars present important differences in relation to their nutritional requirements [4, 5, 8, 17]. Some authors recommend fertilization for groups of cultivars with the same growth habit [8, 17]. In addition, there are research results that also indicate differences between cultivars with and without thorns [5, 21]. But, in general, the greatest difference between demands of each cultivar is actually related to their productive capacity, that is, more productive cultivars export larger quantities of N and therefore also demand higher rates of fertilizer. For this reason, the tendency is that fertilization recommendations incorporate the expected productivity as a criterion to define the most adequate N rate.

#### **6. Nitrogen deficiency**

Compared to blackberry plants with no N deficiency (**Figure 2a**), N deficiency in leaves is characterized by foliar chlorosis (**Figure 2b**), and in severe deficiency situations, reddish patches may appear distributed throughout the leaf blade (**Figure 2c**). In addition to the leaves, the stems may also exhibit reddish pigmentation, and the greater the deficiency, the greater the intensity of the red (**Figure 2e** and **f**), being this type of pigmentation originated from the anthocyanin accumulation [21]. N deficiency appears on old leaves and progresses to the younger ones. This is due to the fact that N is easily translocated and redistributed within the plant [21]. Therefore, when the nutrient supply in the roots is insufficient, the nutrient of the older leaves is mobilized for the younger ones.

The vegetative growth is the major aspect affected by the deficiency of N, being the foliar chlorosis the first visible symptom. This happens because N-deficient plants present a lower chlorophyll leaf concentration, which can be verified by the lower SPAD (soil plant analysis development) index in leaves of nitrogen-deficient plants (**Figure 3a**), which is an indirect measure of the foliar chlorophyll content. It is noted that the SPAD index was not able to identify differences between moderate and severe deficiency. However, the method clearly identified N deficiency even in a moderate situation. In this way, it is possible that through calibration studies, the SPAD index can be used as a rapid method to evaluate the N leaf content in blackberry plants.

In general, when foliar chlorosis is identified, vegetative growth has generally been compromised. Among the main growth parameters that may indicate N deficiency problems in the blackberry are the length of internodes and the length of the stems. In **Figure 3b**, it is possible to observe the average reduction of 15% in the length of the internodes in plants with moderate N deficiency. However, the length of the stems is further reduced. In plants with moderate deficiency, the reduction in stem length was 35%, whereas in plants with severe deficiency, the reduction was 52% (**Figure 3c**).

N deficiency also causes a reduction in the number and in the mass of blackberry fruits. **Figure 3c** shows a reduction of 63% in the number of fruits produced

**61**

**Figure 2.**

*severe nitrogen deficiency (f) [9].*

*Nitrogen Fertilization in Blackberry*

*DOI: http://dx.doi.org/10.5772/intechopen.82409*

in blackberry plants with moderate N deficiency and 65% in plants with severe deficiency. Regarding the fruit mass, the reduction was 52% in plants with moderate

*Leaf superior surface with normal aspect, no deficiency symptoms (a), with moderate (b) and severe nitrogen deficiency (c) in Tupy cultivar. Besides stems without deficiency symptoms (d) and with moderate (e) and*

The increase in fruit size is the main effect of nitrogen fertilization on the fruit quality of blackberries [22]. In general, nitrogen fertilization induces the formation of vigorous stems which, in turn, provide the formation of larger fruits [20]. In terms of sensory quality, the effects are varied. Nitrogen fertilization performed at the recommended amount increases soluble solids concentration but has little effect on attributes such as pH, acidity, sugar/acid ratio, and firmness [23, 24]. On the other hand, the excess of N can cause reduction of soluble solids, increase acidity

The accumulation of negative effects caused by N deficiency on vegetative growth and fruit formation in blackberry plants had a catastrophic impact on plant productivity. It was observed a reduction of 71% in the productivity of plants with

Excess of N is characterized by excessive plant vigor, long internodes, thinner stems, dark green leaves, low yield, low quality of fruits, less conservation potential, and greater risk of diseases [8, 14, 21]. In addition, very high rates of N may result in a decrease in the foliar content of manganese (Mn), potassium (K),

The results presented in **Figure 3** demonstrate that in many aspects there are no significant differences in vegetative growth or fruit yield between plants with moderate and severe N deficiency. This shows that even when N deficiency is moderate,

moderate deficiency and 75% in plants with severe deficiency (**Figure 3f**).

deficiency and 60% when the deficiency was severe (**Figure 3d**).

and pH, and also decrease fruit firmness.

the crop yield is extremely impacted.

#### **Figure 2.**

*Nitrogen Fixation*

the greater the productivity [19].

**6. Nitrogen deficiency**

older leaves is mobilized for the younger ones.

stems have potential for larger fruit production [19, 20].

**5. Nutritional differences between cultivars**

yield of the blackberry, that is, the larger the number of stems (up to 12 stems m−1

On the other hand, postharvest fertilization, usually performed after postharvest pruning, has the function of stimulating the development of primocanes, inducing the formation of vigorous stems and thus capable of supporting high yields and larger fruits in the next season. In blackberry cultivation, there is a significant correlation between stem diameter and fruit size, and more vigorous

It has been observed in the literature that blackberry cultivars present important differences in relation to their nutritional requirements [4, 5, 8, 17]. Some authors recommend fertilization for groups of cultivars with the same growth habit [8, 17]. In addition, there are research results that also indicate differences between cultivars with and without thorns [5, 21]. But, in general, the greatest difference between demands of each cultivar is actually related to their productive capacity, that is, more productive cultivars export larger quantities of N and therefore also demand higher rates of fertilizer. For this reason, the tendency is that fertilization recommendations incorporate

Compared to blackberry plants with no N deficiency (**Figure 2a**), N deficiency in leaves is characterized by foliar chlorosis (**Figure 2b**), and in severe deficiency situations, reddish patches may appear distributed throughout the leaf blade (**Figure 2c**). In addition to the leaves, the stems may also exhibit reddish pigmentation, and the greater the deficiency, the greater the intensity of the red (**Figure 2e** and **f**), being this type of pigmentation originated from the anthocyanin accumulation [21]. N deficiency appears on old leaves and progresses to the younger ones. This is due to the fact that N is easily translocated and redistributed within the plant [21]. Therefore, when the nutrient supply in the roots is insufficient, the nutrient of the

The vegetative growth is the major aspect affected by the deficiency of N, being the foliar chlorosis the first visible symptom. This happens because N-deficient plants present a lower chlorophyll leaf concentration, which can be verified by the lower SPAD (soil plant analysis development) index in leaves of nitrogen-deficient plants (**Figure 3a**), which is an indirect measure of the foliar chlorophyll content. It is noted that the SPAD index was not able to identify differences between moderate and severe deficiency. However, the method clearly identified N deficiency even in a moderate situation. In this way, it is possible that through calibration studies, the SPAD index can be used as a rapid method to evaluate the N leaf content in blackberry plants.

In general, when foliar chlorosis is identified, vegetative growth has generally been compromised. Among the main growth parameters that may indicate N deficiency problems in the blackberry are the length of internodes and the length of the stems. In **Figure 3b**, it is possible to observe the average reduction of 15% in the length of the internodes in plants with moderate N deficiency. However, the length of the stems is further reduced. In plants with moderate deficiency, the reduction in stem length was 35%, whereas in plants with severe deficiency, the reduction was 52% (**Figure 3c**). N deficiency also causes a reduction in the number and in the mass of blackberry fruits. **Figure 3c** shows a reduction of 63% in the number of fruits produced

the expected productivity as a criterion to define the most adequate N rate.

),

**60**

*Leaf superior surface with normal aspect, no deficiency symptoms (a), with moderate (b) and severe nitrogen deficiency (c) in Tupy cultivar. Besides stems without deficiency symptoms (d) and with moderate (e) and severe nitrogen deficiency (f) [9].*

in blackberry plants with moderate N deficiency and 65% in plants with severe deficiency. Regarding the fruit mass, the reduction was 52% in plants with moderate deficiency and 60% when the deficiency was severe (**Figure 3d**).

The increase in fruit size is the main effect of nitrogen fertilization on the fruit quality of blackberries [22]. In general, nitrogen fertilization induces the formation of vigorous stems which, in turn, provide the formation of larger fruits [20]. In terms of sensory quality, the effects are varied. Nitrogen fertilization performed at the recommended amount increases soluble solids concentration but has little effect on attributes such as pH, acidity, sugar/acid ratio, and firmness [23, 24]. On the other hand, the excess of N can cause reduction of soluble solids, increase acidity and pH, and also decrease fruit firmness.

The accumulation of negative effects caused by N deficiency on vegetative growth and fruit formation in blackberry plants had a catastrophic impact on plant productivity. It was observed a reduction of 71% in the productivity of plants with moderate deficiency and 75% in plants with severe deficiency (**Figure 3f**).

The results presented in **Figure 3** demonstrate that in many aspects there are no significant differences in vegetative growth or fruit yield between plants with moderate and severe N deficiency. This shows that even when N deficiency is moderate, the crop yield is extremely impacted.

Excess of N is characterized by excessive plant vigor, long internodes, thinner stems, dark green leaves, low yield, low quality of fruits, less conservation potential, and greater risk of diseases [8, 14, 21]. In addition, very high rates of N may result in a decrease in the foliar content of manganese (Mn), potassium (K),

#### **Figure 3.**

*Effects of different levels of nitrogen (N) deficiency on the SPAD index (a), length of internodes (b), length of stems (c), fruit number per plant (d), fruit mass (e) and yield (f) of 'Tupy' blackberry. Deficiency levels: control or no N deficiency; moderate deficiency, and severe deficiency.*

calcium (Ca), and magnesium (Mg) and increase of copper (Cu). On the other hand, the reduction of fertilization with N reduces the iron content (Fe) and may reduce production due to nutritional imbalance [4, 25]. The reduction of other nutrients can be attributed to two main factors, competition for the same binding sites and the dilution effect, since N stimulates vegetative growth [4].

#### **7. Nitrogen recommendation fertilization**

The fertilization recommendation is divided into **fertilization of preplanting** and **production fertilization** [7]. The preplanting fertilization aims to correct soil problems, as in the case of N, low levels of organic material, and must be carried out during soil preparation, before planting of seedlings, while the production fertilization has the objective to restore to the soil the quantities of N exported by the production of fruits (considers the expectation of production) and natural losses, as, for example, by leaching.

**63**

**Figure 4.**

*Recommendation of nitrogen production fertilization.*

*Nitrogen Fertilization in Blackberry*

**7.1 Preplanting fertilization**

**7.2 Production fertilization**

expectation (**Figure 4**).

*DOI: http://dx.doi.org/10.5772/intechopen.82409*

Nitrogen fertilization of preplanting is recommended in soils with organic matter content of less than 2.5%. In these cases, it is recommended to apply 40 kg ha−1 of N, and it is preferable to use organic sources, such as compost from manure.

Production fertilization begins in the year following to planting of the seedlings

and takes into account soil organic matter content, plant age, and production

The N rate applied in the spring should be divided into three times: the first application should be made at the beginning of the budding; the second, 30 days

#### **7.1 Preplanting fertilization**

*Nitrogen Fixation*

**62**

**Figure 3.**

calcium (Ca), and magnesium (Mg) and increase of copper (Cu). On the other hand, the reduction of fertilization with N reduces the iron content (Fe) and may reduce production due to nutritional imbalance [4, 25]. The reduction of other nutrients can be attributed to two main factors, competition for the same binding

*Effects of different levels of nitrogen (N) deficiency on the SPAD index (a), length of internodes (b), length of stems (c), fruit number per plant (d), fruit mass (e) and yield (f) of 'Tupy' blackberry. Deficiency levels:*

The fertilization recommendation is divided into **fertilization of preplanting**

and **production fertilization** [7]. The preplanting fertilization aims to correct soil problems, as in the case of N, low levels of organic material, and must be carried out during soil preparation, before planting of seedlings, while the production fertilization has the objective to restore to the soil the quantities of N exported by the production of fruits (considers the expectation of production)

sites and the dilution effect, since N stimulates vegetative growth [4].

**7. Nitrogen recommendation fertilization**

*control or no N deficiency; moderate deficiency, and severe deficiency.*

and natural losses, as, for example, by leaching.

Nitrogen fertilization of preplanting is recommended in soils with organic matter content of less than 2.5%. In these cases, it is recommended to apply 40 kg ha−1 of N, and it is preferable to use organic sources, such as compost from manure.

#### **7.2 Production fertilization**

Production fertilization begins in the year following to planting of the seedlings and takes into account soil organic matter content, plant age, and production expectation (**Figure 4**).

The N rate applied in the spring should be divided into three times: the first application should be made at the beginning of the budding; the second, 30 days

**Figure 4.** *Recommendation of nitrogen production fertilization.*

#### **Table 1.**

*Tissue N concentration from sampling after harvesting the primocane's leaves [8].*

after; and the third, at 60 days after the first application. The application postharvest can be applied in a single rate.

The N may be applied in the form of urea (45% of N), calcium nitrate (14% of N), ammonium nitrate (32% of N), or ammonium sulfate (20% of N). Ammonium sulfate is preferably recommended because the blackberries require sulfur. However, the consecutive use of ammonium sulfate can significantly reduce soil pH, being necessary to monitor this parameter. An alternative to supply the sulfur demand with little impact on soil pH may be the use of agricultural gypsum (CaSO4.2H2O), which also provides calcium and can be applied at a rate of 34–45 kg ha−1 to supply possible S deficiencies of blackberries [8].

The N source application should be performed at the soil surface, along the planting row, approximately 15 cm away from crown of plants.

#### **7.3 Leaf N concentrations**

In contrast to other species, in blackberry, leaf analysis is applied to evaluate the nutritional status after harvest, aiming the elaboration of a fertilization program for the next productive cycle (**Table 1**). For this reason, sampling is performed after harvesting the primocane's leaves. Sampling is performed on the 6th completely expanded leave from the stem apex.

#### **8. Conclusion**

Nutrition studies for blackberry crop are scarce worldwide. However, as presented in this chapter, nitrogen fertilization is one aspect of crop management that has the greatest impact on vegetative growth, fruit yield, and quality. It has been demonstrated in this chapter that even under conditions of moderate N deficiency, there can be significant impacts on blackberry yield. Nitrogen-deficient plants tend to form weak, low-yielding stems. In this way, the application of adequate rates for each type of soil and cultivar is fundamental for satisfactory productivities.

In addition, there are important differences in the demand of N between cultivars, probably due to productive capacity, that is, cultivars that provide higher yields also export larger amounts of N and therefore need larger amount of nitrogen fertilizers.

In this way, a nitrogen fertilization program was proposed, based on SOM (indicator of N availability), plant age, and production expectation.

#### **Acknowledgements**

The authors are grateful to the Brazilian Agricultural Research Corporation (EMBRAPA) and the Soil and Water Management and Conservation Post-Graduate Program at the Federal University of Pelotas (UFPel) for the financial and infrastructure support.

**65**

**Author details**

Ivan dos Santos Pereira1

Luis Eduardo Corrêa Antunes2

\*, Adilson Luis Bamberg2

2 Brazilian Agricultural Research Corporation (EMBRAPA), Pelotas, Brazil

© 2019 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,

1 Federal University of Pelotas (UFPel), Pelotas, Brazil

\*Address all correspondence to<EMAIL_ADDRESS>
provided the original work is properly cited.

and Rogério Oliveira de Sousa1

, Carlos Augusto Posser Silveira2

,

*Nitrogen Fertilization in Blackberry*

*DOI: http://dx.doi.org/10.5772/intechopen.82409*

*Nitrogen Fertilization in Blackberry DOI: http://dx.doi.org/10.5772/intechopen.82409*

*Nitrogen Fixation*

**Table 1.**

34–45 kg ha−1

**8. Conclusion**

**Acknowledgements**

and infrastructure support.

**7.3 Leaf N concentrations**

expanded leave from the stem apex.

vest can be applied in a single rate.

after; and the third, at 60 days after the first application. The application posthar-

The N may be applied in the form of urea (45% of N), calcium nitrate (14%

 to supply possible S deficiencies of blackberries [8]. The N source application should be performed at the soil surface, along the

In contrast to other species, in blackberry, leaf analysis is applied to evaluate the nutritional status after harvest, aiming the elaboration of a fertilization program for the next productive cycle (**Table 1**). For this reason, sampling is performed after harvesting the primocane's leaves. Sampling is performed on the 6th completely

Nutrition studies for blackberry crop are scarce worldwide. However, as presented in this chapter, nitrogen fertilization is one aspect of crop management that has the greatest impact on vegetative growth, fruit yield, and quality. It has been demonstrated in this chapter that even under conditions of moderate N deficiency, there can be significant impacts on blackberry yield. Nitrogen-deficient plants tend to form weak, low-yielding stems. In this way, the application of adequate rates for

In addition, there are important differences in the demand of N between cultivars, probably due to productive capacity, that is, cultivars that provide higher yields also export larger amounts of N and therefore need larger amount of nitrogen fertilizers. In this way, a nitrogen fertilization program was proposed, based on SOM

The authors are grateful to the Brazilian Agricultural Research Corporation

(EMBRAPA) and the Soil and Water Management and Conservation Post-Graduate Program at the Federal University of Pelotas (UFPel) for the financial

each type of soil and cultivar is fundamental for satisfactory productivities.

(indicator of N availability), plant age, and production expectation.

planting row, approximately 15 cm away from crown of plants.

of N), ammonium nitrate (32% of N), or ammonium sulfate (20% of N). Ammonium sulfate is preferably recommended because the blackberries require sulfur. However, the consecutive use of ammonium sulfate can significantly reduce soil pH, being necessary to monitor this parameter. An alternative to supply the sulfur demand with little impact on soil pH may be the use of agricultural gypsum (CaSO4.2H2O), which also provides calcium and can be applied at a rate of

**Class Nitrogen content (%)**

Insufficient <2.20 Normal 2.30–3.00 Excessive >3.00

*Tissue N concentration from sampling after harvesting the primocane's leaves [8].*

**64**

## **Author details**

Ivan dos Santos Pereira1 \*, Adilson Luis Bamberg2 , Carlos Augusto Posser Silveira2 , Luis Eduardo Corrêa Antunes2 and Rogério Oliveira de Sousa1

1 Federal University of Pelotas (UFPel), Pelotas, Brazil

2 Brazilian Agricultural Research Corporation (EMBRAPA), Pelotas, Brazil

\*Address all correspondence to<EMAIL_ADDRESS>
© 2019 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.

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[3] Grandall PC. Bramble Production: The Management and Marketing of Raspberries and Blackberries. 1st ed. Binghamton, N.Y.: CRC Press; 1995. 172 p

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[5] Pereira IS, Silveira CAP, Picolotto L, Schneider FC, Gonçalves MA, Vignolo GK, et al. Constituição química e exportação de nutrientes da amoreirapreta. Revista Congrega URCAMP. 2013;**9**:1-10

[6] Castaño CA, Morales CS, Obando FH. Evaluación de las deficiencias nutricionales em el cultivo de la mora (*Rubus glaucus*) en condiciones controladas para bosque montano bajo. Agronomía. 2008;**16**:75-88

[7] CQFS. Comissão de Química e Fertilidade do Solo—RS/SC. Manual de calagem e adubação para os estados do Rio Grande Sul e Santa Catarina. 11ª ed. SBCS-NRS: Porto Alegre; 2016. 376 p

[8] Hart J, Strik B, Rempel H. Caneberries. In: Nutrient Management. Oregon State University; January 2006. 8 p

[9] Pereira IS, Nava G, Picolotto L, Silveira CAP, Vignolo GK, Gonçalves MA, et al. Exigência nutricional e adubação da amoreirapreta: Nutritional requirements and

blackberry fertilization. Revista de Ciências Agrárias. 2015;**58**:96-104. DOI: 10.4322/rca.1755

[10] Lawson HM, Waister PD. The response to nitrogen of a raspberry plantation under contrasting systems of management for a weed and sucker control. Horticulture Research. 1972;**12**:43-55

[11] Bushway L, Pritts M, Handley D, editors. Raspberry & Blackberry Production Guide for the Northeast, Midwest, and Eastern Canada. Plant and Life Sciences Publishing Coop.; 2008. 157 p. Ext. NRAES-35

[12] Gutser RT, Ebertseder A, Weber M, Schraml, Schmidhalter U. Short-term and residual availability of nitrogen after long-term application of organic fertilizers on arable land. Journal of Plant Nutrition and Soil Science. 2005;**168**:439-445

[13] Kowalenko CG, Keng JCW, Freeman JA. Comparison of nitrogen application via a trickle irrigation system with surface banding of granular fertilizer on red raspberry. Canadian Journal of Plant Science. 2000;**80**:363- 371. DOI: 10.4141/P99-094

[14] Freire CJS. Nutrição e adubação. In: Cultivo da amoreira-preta. In: Antunes LEC, Raseira MCB, editors. Sistemas de Produção. Pelotas; 2007. pp. 45-54

[15] Pereira IS. Adubação de préplantio no crescimento, produção e qualidade da amoreira-preta (*Rubus* sp.) [Dissertação]. Pelotas: Universidade Federal de Pelotas; 2008

[16] Pritts M, Handley D. Bramble Production. Ithaca, NY: Northeast Regional Agricultural Engineering Service Pub. 35; 1989

**67**

*Nitrogen Fertilization in Blackberry*

2008;(777):403-410

[17] Strik BC. A review of nitrogen nutrition of Rubus. Acta Horticulturae.

[18] Dickerson GW, editor. Blackberry Production in New Mexico. Cooperative

Agriculture and Home Economics, New Mexico State University (NMSU) and the U.S. Department of Agriculture Cooperating, Guide H-325; 2000. 8 p

Extension Service College of

[19] Pereira IS, Antunes LEC,

[20] Eyduran SP, Eyduran E,

[21] Antunes LEC, Pereira IS, Picolotto L, Vignolo GK,

10.1590/0100-2945-450/13

[22] Pereira IS, Antunes LEC,

[23] Nelson E, Martin LW. The relationship of soil-applied N and K to yield and quality of 'Thornless Evergreen' blackberry. HortScience.

[24] Alleyne V, Clark JR. Fruit composition of 'Arapaho' blackberry following nitrogen fertilization. HortScience. 1997;**32**:282-283

1986;**21**:1153-1154

Silveira CAP, Messias RS, Gardin JPP, Schneider FC, et al. Caracterização agronômica da amoreira-preta cultivada no sul do estado do Paraná. Pelotas: Embrapa Clima Temperado; 2009. 19 p

Agaoglu YS. Estimation of fruit weight by cane traits for eight American blackberries (*Rubus fruticosus* L.) cultivars. African Journal of Biotechnology. 2008;**7**:3031-3038

Gonçalves MA. Produção de amoreirapreta no Brasil. Revista Brasileira de Fruticultura. 2014;**36**:100-111. DOI:

Messias RS, Silveira CAP, Vignolo GK. Avaliações da subtração dos elementos N, P e K sobre a produção e qualidade de frutos de amoreira-preta. In: Comunicado Técnico 275. Pelotas: Embrapa Clima Temperado; 2012. 6 p

*DOI: http://dx.doi.org/10.5772/intechopen.82409*

[25] Spiers JM, Braswell JH. Influence of N, P, K, Ca, and Mg rates on leaf macronutrient concentration of

2002;(585):659-663

'Navaho' blackberry. Acta Horticulturae.

*Nitrogen Fertilization in Blackberry DOI: http://dx.doi.org/10.5772/intechopen.82409*

[17] Strik BC. A review of nitrogen nutrition of Rubus. Acta Horticulturae. 2008;(777):403-410

[18] Dickerson GW, editor. Blackberry Production in New Mexico. Cooperative Extension Service College of Agriculture and Home Economics, New Mexico State University (NMSU) and the U.S. Department of Agriculture Cooperating, Guide H-325; 2000. 8 p

[19] Pereira IS, Antunes LEC, Silveira CAP, Messias RS, Gardin JPP, Schneider FC, et al. Caracterização agronômica da amoreira-preta cultivada no sul do estado do Paraná. Pelotas: Embrapa Clima Temperado; 2009. 19 p

[20] Eyduran SP, Eyduran E, Agaoglu YS. Estimation of fruit weight by cane traits for eight American blackberries (*Rubus fruticosus* L.) cultivars. African Journal of Biotechnology. 2008;**7**:3031-3038

[21] Antunes LEC, Pereira IS, Picolotto L, Vignolo GK, Gonçalves MA. Produção de amoreirapreta no Brasil. Revista Brasileira de Fruticultura. 2014;**36**:100-111. DOI: 10.1590/0100-2945-450/13

[22] Pereira IS, Antunes LEC, Messias RS, Silveira CAP, Vignolo GK. Avaliações da subtração dos elementos N, P e K sobre a produção e qualidade de frutos de amoreira-preta. In: Comunicado Técnico 275. Pelotas: Embrapa Clima Temperado; 2012. 6 p

[23] Nelson E, Martin LW. The relationship of soil-applied N and K to yield and quality of 'Thornless Evergreen' blackberry. HortScience. 1986;**21**:1153-1154

[24] Alleyne V, Clark JR. Fruit composition of 'Arapaho' blackberry following nitrogen fertilization. HortScience. 1997;**32**:282-283

[25] Spiers JM, Braswell JH. Influence of N, P, K, Ca, and Mg rates on leaf macronutrient concentration of 'Navaho' blackberry. Acta Horticulturae. 2002;(585):659-663

**66**

*Nitrogen Fixation*

**References**

[1] Taiz L, Zeiger E. Fisiologia Vegetal. 5th ed. Artmed: Porto Alegre; 2013. 820 p

blackberry fertilization. Revista de Ciências Agrárias. 2015;**58**:96-104. DOI:

[10] Lawson HM, Waister PD. The response to nitrogen of a raspberry plantation under contrasting systems of management for a weed and sucker control. Horticulture Research.

[11] Bushway L, Pritts M, Handley D, editors. Raspberry & Blackberry Production Guide for the Northeast, Midwest, and Eastern Canada. Plant and Life Sciences Publishing Coop.;

[12] Gutser RT, Ebertseder A, Weber M, Schraml, Schmidhalter U. Short-term and residual availability of nitrogen after long-term application of organic fertilizers on arable land. Journal of Plant Nutrition and Soil Science.

2008. 157 p. Ext. NRAES-35

[13] Kowalenko CG, Keng JCW, Freeman JA. Comparison of nitrogen application via a trickle irrigation system with surface banding of granular fertilizer on red raspberry. Canadian Journal of Plant Science. 2000;**80**:363-

371. DOI: 10.4141/P99-094

[14] Freire CJS. Nutrição e adubação. In: Cultivo da amoreira-preta. In: Antunes LEC, Raseira MCB, editors. Sistemas de Produção. Pelotas; 2007.

[15] Pereira IS. Adubação de préplantio no crescimento, produção e qualidade da amoreira-preta (*Rubus* sp.) [Dissertação]. Pelotas: Universidade

[16] Pritts M, Handley D. Bramble Production. Ithaca, NY: Northeast Regional Agricultural Engineering

Federal de Pelotas; 2008

Service Pub. 35; 1989

2005;**168**:439-445

pp. 45-54

10.4322/rca.1755

1972;**12**:43-55

[2] Chapin FS. The mineral nutrition of wild plants. Annual Review of Ecology and Systematics. 1980;**11**:233-260

[3] Grandall PC. Bramble Production: The Management and Marketing of Raspberries and Blackberries. 1st ed. Binghamton, N.Y.: CRC Press; 1995. 172 p

[4] Pereira IS, Picolotto L, Messias RS, Potes ML, Antunes LEC. Adubação nitrogenada e características agronômicas em amoreira-preta. Pesquisa Agropecuária Brasileira. 2013;**48**:373-380. DOI: 10.1590/ S0100-204X2013000400004

[5] Pereira IS, Silveira CAP, Picolotto L,

Vignolo GK, et al. Constituição química e exportação de nutrientes da amoreirapreta. Revista Congrega URCAMP.

Schneider FC, Gonçalves MA,

[6] Castaño CA, Morales CS, Obando FH. Evaluación de las deficiencias nutricionales em el cultivo de la mora (*Rubus glaucus*) en condiciones controladas para bosque montano bajo. Agronomía.

[7] CQFS. Comissão de Química e Fertilidade do Solo—RS/SC. Manual de calagem e adubação para os estados do Rio Grande Sul e Santa Catarina. 11ª ed. SBCS-NRS: Porto Alegre; 2016. 376 p

University; January 2006. 8 p

Silveira CAP, Vignolo GK, Gonçalves MA, et al. Exigência

Nutritional requirements and

[9] Pereira IS, Nava G, Picolotto L,

[8] Hart J, Strik B, Rempel H. Caneberries. In: Nutrient Management. Oregon State

nutricional e adubação da amoreirapreta:

2013;**9**:1-10

2008;**16**:75-88

**Chapter 5**

**Abstract**

**1. Introduction**

**69**

Nitrogen Fertilization I: Impact on

*Upendra M. Sainju, Rajan Ghimire and Gautam P. Pradhan*

Nitrogen (N) is a major limiting nutrient to sustain crop yields and quality. As a result, N fertilizer is usually applied in large quantity to increase crop production throughout the world. Application of N fertilizers has increased crop yields and resulted in achievement of self-sufficiency in food production in many developing countries. Excessive application of N fertilizers beyond crops' demand, however, has resulted in undesirable consequences of degradation in soil, water, and air quality. These include soil acidification, N leaching in groundwater, and emissions of nitrous oxide (N2O), a potent greenhouse gas that contributes to global warming. Long-term application of ammonia-based N fertilizers, such as urea, has increased soil acidity which rendered to soil infertility where crops fail to respond with further application of N fertilizers. Another problem is the groundwater contamination of nitrate-N (NO3-N) which can be a health hazard to human and livestock if its concentration goes above 10 mg L1 in drinking water. The third problem is emissions of N2O gas which is 300 times more powerful than carbon dioxide in terms of global warming potential. This chapter examines the effect of N fertiliza-

Crop, Soil, and Environment

tion on soil and environmental quality and crop yields.

nitrogen fertilizer, nitrogen-use efficiency, soil quality

**Keywords:** crop yields, environmental quality, management practices,

Nitrogen (N) is a major limiting factor for sustainable and profitable crop production. However, excessive N application through fertilizers and manures can degrade soil and environmental quality by increasing soil acidification, N leaching, and emissions of ammonia (NH3) and nitrogen oxide (NO, N2O, and NO2) gases, out of which N2O is considered a highly potent greenhouse gas that contributes to global warming [1, 2]. Nitrogen application more than crop's need can also result in

reduced yield [3]. Additional N inputs include dry and wet (snow and rain) depositions from the atmosphere, biological N fixation, and irrigation water. Because crops can remove about 40–60% of applied N, the soil residual N

(nitrate-N [NO3-N] + ammonium-N [NH4-N]) after crop harvest can be lost to the environment through leaching, denitrification, volatilization, surface runoff, soil erosion, and N2O emissions [3, 4]. One option to reduce soil residual N is to increase N-use efficiency. Nitrogen-use efficiency for crops, however, can be lower at high N fertilization rates [5]. Improved management practices can increase N-use efficiency, enhance soil N storage, and reduce N fertilizer application which

#### **Chapter 5**


*2.3. Materials*

In this study, a 12 μg/h transdermal fentanyl patch (Fentanyl-Mepha Matrixpfl, Mepha Pharma AG, Switzerland) was used. Due to the recommendation of a dosage of 2 μg/kg/h in sheep and up to 5 μg/kg/h in small animals (dogs, cats) described in several former studies, the 12 μg/h patch size was chosen [5,7,22]. The mean administered dose rate of fentanyl was 3.60 μg/kg/h (Dmax = 3.93 μg/kg/h, Dmin = 3.16 μg/kg/h).

### *2.4. Fentanyl Patch Application Process*

Rabbits were sedated with Medetomidin (200 μg/kg) and Midazolam (0.5 mg/kg) both mixed in a syringe and given intramuscularly approximately 15 min before starting skin preparation. Thereafter, the rabbits were placed in sternal recumbency, eye ointment (Vitamin A Blache Augensalbe, Bausch & Lomb Swiss AG, Switzerland) was applied and oxygen (flow rate 1 L/min) was provided via face mask. The required site was carefully clipped using two different clippers depending on the location. For the location on the ear, the clipper Isis (GT420 Aesculap, Germany; cutting length 0.5 mm) was utilized. For the neck, another mechanical fur clipper (model GH703/10 (Favorita II GT104), Aesculap, Germany; cutting length 0.1 mm) was used in the interscapular region due to the increased density of hair at this site. During clipping extreme precautions were taken not to traumatize the application site to avoid influencing fentanyl absorption.

Afterwards, skin was degreased with swabs (Mesoft, Moelnlycke, Sweden) soaked in alcohol (Softasept, B. Braun Vet Care GmbH, Germany) to ensure a good contact of the patch with the skin. While waiting for 5 min to air dry, the ear not used for patch application was clipped and prepared aseptically with alcohol for the placement of a 22 G catheter (Vasofix Safety, B.Braun Meisungen AG, Germany) in the marginal ear vein. The catheter, inserted to facilitate blood withdrawal, was wrapped with tape (Durapore, 3 M (Schweiz) GmbH, Switzerland) and a roll of gauze (Mesoft) to hold it in place. After each blood withdrawal, the catheter was flushed with approximately 0.5 mL of heparinized 0.9% NaCl (B.Braun Meisungen AG, Germany) and a mandrin (Mandrin Vasofix, B.Braun Meisungen AG, Germany) was inserted. On the dry application site, the sticky side of the patch was then pressed on the skin with the palm of the hand and kept there for 1 min to ensure a good patch-skin contact. Finally, the patch was fixed with tape (Leukoplast) to prevent the loosening of the patch detachment during animal handling.

### *2.5. Sample Collection and Plasma Fentanyl Analysis via ELISA*

Venous blood sampling was performed immediately prior to the application of the patch and 3, 6, 9, 12, 18, 24, 36, 48, 72, 96, 120 h thereafter. In a few cases arterial blood was sampled, if no venous sample could be obtained. At each sampling time point, approximately 1 mL of blood was collected in a 1.3 mL EDTA-covered tube (1.3 mL K3E, Sarstedt, AG & Co, Nümbrecht, Germany). A venous catheter was always used to withdraw blood samples. For the first 24 h the catheter was left in the marginal ear vein. Thereafter, a new catheter was inserted for each blood sampling. EDTA-tubes were then centrifuged for 15 min, at 23 ◦C and 1000 rcf (Centrifuge 5810R, Vaudaux-Eppendorf AG, Switzerland) within 2 h of collection.

Subsequently, the obtained plasma was transferred into Eppendorf tubes (Vaudaux-Eppendorf AG, Switzerland) and stored at −80 ◦C until analysis. Plasma fentanyl concentrations were measured by using a commercially available human enzyme-linked immunoabsorbent assay (ELISA). Fentanyl Kits (Adnova, Fentanyl (Human) ELISA Kit, Taiwan) were used according to the manufacturer's instructions. Additionally, serial dilutions of fentanyl standard (Fentanyl Sintetica 0.5 mg/10 mL) in fentanyl negative rabbit plasma were measured on the ELISA-plate, as a standard curve. The absorbance was read at 450 nm using a Mithras microplate reader (Berthold Technologies, Bad Wildbad, Germany). Each sample was tested in duplicates.

### *2.6. Scoring and Practicability*

During the study, the rabbits were scored twice a day. The scoring system included observation of food and water intake, assessment of the general conditions, body weight and rectal temperature as well as defecation and coprophagy. Additionally, practicability was assessed based on the following criteria: ease of preparation, quality of patch adhesiveness, ease of daily checks, occurrence of undesired patch detachment before the end of the study, ease of patch removal and skin condition after patch removal. Any findings in respect to these criteria were noted and at the end a subjective three level scoring system (positive, neutral and negative assessment) comparing the three groups was applied by one veterinarian.