Amelioration of Drought Stress on Plants under Biostimulant Sources

*Ana Carolina Feitosa de Vasconcelos*

### **Abstract**

Water stress is one of the most important environmental factors inducing physiological changes in plants, such as decreasing water potential of the cells and the stomatal closure, resulting in reduced CO2 availability for the plants and inhibiting photosynthesis. One common feature of these stress conditions is the development of oxidative processes mediated by reactive oxygen species (ROS). ROS accumulate in the cells and cause damage in important cellular components, such as thylakoids and chloroplasts. Plants have antioxidant defense systems to cope with ROS. Antioxidants enzymes superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) are efficient scavengers of ROS: superoxide, hydroxyl radicals, and singlet oxygen. The activities of antioxidant enzymes in plants are normally favored when plants are subjected to some kind of improvement in the conditions in which they are grown. In this sense, biostimulants cause changes in vital and structural processes in order to influence plant growth through improved tolerance to abiotic stresses by increasing the antioxidant activity in plants.

**Keywords:** reactive oxygen species, antioxidant enzymes, plant drought resistance, humic substances, seaweed extracts, hormones

### **1. Introduction**

Water availability is one of the most important environmental factors for plant growth and development. The water deficit caused by drought or salinity in soils is one of the most serious environmental problems that limit agricultural production in various regions of the world. According to [1], water deficit occurs when all water content in the cell is below the highest water content displayed in the state of greatest hydration.

Plants experience a water deficit when water supply to the roots becomes difficult or when the rate of evapotranspiration becomes very high. These two conditions generally coincide in regions with an arid and semiarid climate and affect plants to a greater or lesser extent according to the tolerance that species have [2].

Plant response to biotic and abiotic stresses is a complex network of reactions, which involves different physiological pathways of the primary and secondary metabolism. At the cellular level, membranes and proteins can be damaged by a reduction in hydration and an increase in reactive oxygen species (ROS) [3]. ROS derive from oxidative processes such as photosynthesis and respiration, and, in normal conditions, they are produced in low concentration without any negative consequences for the plants. In stressful conditions (biotic or abiotic), ROS levels

increase as an index of the oxidative burst induced by the stress agent [4]. When ROS become toxic, they can result in a series of damages to plant metabolism, such as deterioration of photosynthetic components, inactivation of proteins and enzymes, and destruction of the structure and permeability of the cell membrane by lipid peroxidation [5, 6].

Antioxidants and their role in the plant defense system have received a lot of attention in scientific research. Many results suggest that the effects of environmental stresses, such as salinity, drought, low temperatures, and herbicide residues, damage plants directly or indirectly by increasing endogenous ROS [7].

Plant cells are protected against the damaging effects of ROS by a complex antioxidant system composed of enzymatic antioxidants, such as superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) [8]. The close relationship between antioxidant activity and stress tolerance has been identified in many crops such as maize (*Zea mays* L.) [7], tobacco (*Nicotiana tabacum*) [9], and grasses [10].

Biostimulants are extracts obtained from organic raw materials containing bioactive compounds. The most common components of the biostimulants are mineral elements, humic substances (HSs), vitamins, and amino acids [6]. Seaweed extracts have been used in agriculture as soil conditioners or as plant stimulators. They are applied as foliar spray and enhance plant growth; freezing, drought, and salt tolerance; photosynthetic activity; and resistance to fungi, bacteria, and virus, improving the yield and productivity of many crops [11, 12]. Seaweeds used for biostimulant production contain cytokinins and auxins or other hormone-like substances [13]. From a legal point of view, the biostimulants can contain traces of natural plant hormones, but their biological action should not be ascribed to them; otherwise they should be registered as plant growth regulators [6].

Humic acids have been used in the composition of many commercial products because they have phytohormones [14] that favor protection against oxidative damage in plants caused by environmental stresses. Thus, the use of biostimulants in agriculture has been emphasized, which are products that contain active ingredient or organic agent free of pesticides, capable of acting, directly or indirectly, on all or part of the cultivated plants, increasing their productivity [15].

The components of biostimulants can change the hormonal status of the plant and have a great influence on its development and health. Seaweed, humic acids, and vitamins are commonly present in biostimulants and are important in improving plant development and hormonal activity [16]. In addition, these products increase the antioxidant activity in plants, especially when they are under water stress, severe temperatures, and herbicide action, among others [7].

Several studies have shown results in improving the resistance of plants to water stress when subjected to the application of biostimulants. The activity levels of the antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) have been determined. In general, increases in these antioxidant enzymes have been observed with the use of biostimulants [16]. Another parameter that has been improved in the plant with the application of biostimulants is the photochemical efficiency [17].

Thus, the objective of this chapter was to approach the role of biostimulants in plants submitted to water supply deficit, by affecting the activities of enzymatic antioxidants.

### **2. Use of biostimulants in plants**

Biostimulants are components that produce responses in plant growth by improving tolerance to abiotic stresses. Many of the effects of these products are

**339**

summer [23].

*Amelioration of Drought Stress on Plants under Biostimulant Sources*

the plant by increasing its level of antioxidant enzymes [15].

temperatures and herbicide action, among others [20].

based on their ability to influence the hormonal activity of plants. Phytohormones are chemical messengers that regulate the normal development of plants by growing roots and shoots, in addition to regulating responses to the environment where

Many statements about biostimulants also refer to the improvements they provide in the tolerance of plants to water stress, a limiting factor in the management of the crops. Water stress affects many metabolic functions in plants, specifically photosynthesis. The application of biostimulants increases the defense system of

The components of biostimulants can alter the plant's hormonal status and have a major influence on its growth and health. Seaweed, humic acids and vitamins are commonly present in biostimulants and are important in improving plant development and hormonal activity [19]. In addition, these products increase the antioxidant activity in plants, especially when they are under water stress, severe

However, the composition of biostimulants is partly unknown; the complexity of the extracts and the wide range of molecules contained in the solution make it very difficult to understand which the most active compounds are. Moreover, the isolation and study of a single component present in a biostimulant can produce unreliable results because the effects on plants are often due to the combination and synergistic action of different compounds. In addition, the mechanisms activated

Plants usually thrive when the environment is favorable. Under these conditions, the effects of biostimulants may not be easily identified. However, when plants are stressed and undergo treatment with biostimulants, they develop better, as their defense system becomes more efficient due to the increase in their levels of antioxidants [20]. Besides, many of the active substances of biostimulants can be present in very low concentrations, sometimes below the levels detectable with commonly available technologies, but can provide strong

Biostimulants and humic substances have shown an influence on many metabolic processes in plants, such as respiration, photosynthesis, synthesis of nucleic acids, and ion absorption. Within the cell, humic substances can increase the chlorophyll content resulting in greener leaves and reduction of some problems in plants, such as leaf chlorosis, since humic substances improve the capacity of nutrient uptake by the roots [21]. Beyond humic substances, various raw materials have been used in biostimulant compositions, such as hormones, algae extracts, and

Water availability is one of the most limiting environmental factors that affect crop productivity. In the semiarid tropics, the occurrence of drought or water deficit in the soil is quite common, despite the fact that crops in regions of tropical and temperate climate suffer seasonal periods of water deficit, especially during the

Drought is a prevalent stress factor especially in arid and semiarid areas and can affect different aspects of plant growth, development, and metabolism. Drought is a multidimensional stress factor, and hence its effects on plants are complex. Its effects on plants can occur on a molecular level up to a whole-plant level. There are several reasons for drought in nature, including low rainfall, salinity, high

temperature, and high intensity of light, among others [24].

by biostimulants are difficult to identify and still under investigation [6].

*DOI: http://dx.doi.org/10.5772/intechopen.91975*

they are located [18].

biological effects [6].

plant growth-promoting bacteria [22].

**3. Water stress in plants**

### *Amelioration of Drought Stress on Plants under Biostimulant Sources DOI: http://dx.doi.org/10.5772/intechopen.91975*

*Plant Stress Physiology*

lipid peroxidation [5, 6].

increase as an index of the oxidative burst induced by the stress agent [4]. When ROS become toxic, they can result in a series of damages to plant metabolism, such as deterioration of photosynthetic components, inactivation of proteins and enzymes, and destruction of the structure and permeability of the cell membrane by

Antioxidants and their role in the plant defense system have received a lot of attention in scientific research. Many results suggest that the effects of environmental stresses, such as salinity, drought, low temperatures, and herbicide residues,

Plant cells are protected against the damaging effects of ROS by a complex antioxidant system composed of enzymatic antioxidants, such as superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) [8]. The close relationship between antioxidant activity and stress tolerance has been identified in many crops such as maize (*Zea mays* L.) [7], tobacco (*Nicotiana tabacum*) [9], and grasses [10]. Biostimulants are extracts obtained from organic raw materials containing bioactive compounds. The most common components of the biostimulants are mineral elements, humic substances (HSs), vitamins, and amino acids [6]. Seaweed extracts have been used in agriculture as soil conditioners or as plant stimulators. They are applied as foliar spray and enhance plant growth; freezing, drought, and salt tolerance; photosynthetic activity; and resistance to fungi, bacteria, and virus, improving the yield and productivity of many crops [11, 12]. Seaweeds used for biostimulant production contain cytokinins and auxins or other hormone-like substances [13]. From a legal point of view, the biostimulants can contain traces of natural plant hormones, but their biological action should not be ascribed to them;

Humic acids have been used in the composition of many commercial products because they have phytohormones [14] that favor protection against oxidative damage in plants caused by environmental stresses. Thus, the use of biostimulants in agriculture has been emphasized, which are products that contain active ingredient or organic agent free of pesticides, capable of acting, directly or indirectly, on all or

The components of biostimulants can change the hormonal status of the plant and have a great influence on its development and health. Seaweed, humic acids, and vitamins are commonly present in biostimulants and are important in improving plant development and hormonal activity [16]. In addition, these products increase the antioxidant activity in plants, especially when they are under water

Several studies have shown results in improving the resistance of plants to water stress when subjected to the application of biostimulants. The activity levels of the antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) have been determined. In general, increases in these antioxidant enzymes have been observed with the use of biostimulants [16]. Another parameter that has been improved in the plant with the application of biostimulants is the

Thus, the objective of this chapter was to approach the role of biostimulants in plants submitted to water supply deficit, by affecting the activities of enzymatic

Biostimulants are components that produce responses in plant growth by improving tolerance to abiotic stresses. Many of the effects of these products are

damage plants directly or indirectly by increasing endogenous ROS [7].

otherwise they should be registered as plant growth regulators [6].

part of the cultivated plants, increasing their productivity [15].

stress, severe temperatures, and herbicide action, among others [7].

**338**

antioxidants.

photochemical efficiency [17].

**2. Use of biostimulants in plants**

based on their ability to influence the hormonal activity of plants. Phytohormones are chemical messengers that regulate the normal development of plants by growing roots and shoots, in addition to regulating responses to the environment where they are located [18].

Many statements about biostimulants also refer to the improvements they provide in the tolerance of plants to water stress, a limiting factor in the management of the crops. Water stress affects many metabolic functions in plants, specifically photosynthesis. The application of biostimulants increases the defense system of the plant by increasing its level of antioxidant enzymes [15].

The components of biostimulants can alter the plant's hormonal status and have a major influence on its growth and health. Seaweed, humic acids and vitamins are commonly present in biostimulants and are important in improving plant development and hormonal activity [19]. In addition, these products increase the antioxidant activity in plants, especially when they are under water stress, severe temperatures and herbicide action, among others [20].

However, the composition of biostimulants is partly unknown; the complexity of the extracts and the wide range of molecules contained in the solution make it very difficult to understand which the most active compounds are. Moreover, the isolation and study of a single component present in a biostimulant can produce unreliable results because the effects on plants are often due to the combination and synergistic action of different compounds. In addition, the mechanisms activated by biostimulants are difficult to identify and still under investigation [6].

Plants usually thrive when the environment is favorable. Under these conditions, the effects of biostimulants may not be easily identified. However, when plants are stressed and undergo treatment with biostimulants, they develop better, as their defense system becomes more efficient due to the increase in their levels of antioxidants [20]. Besides, many of the active substances of biostimulants can be present in very low concentrations, sometimes below the levels detectable with commonly available technologies, but can provide strong biological effects [6].

Biostimulants and humic substances have shown an influence on many metabolic processes in plants, such as respiration, photosynthesis, synthesis of nucleic acids, and ion absorption. Within the cell, humic substances can increase the chlorophyll content resulting in greener leaves and reduction of some problems in plants, such as leaf chlorosis, since humic substances improve the capacity of nutrient uptake by the roots [21]. Beyond humic substances, various raw materials have been used in biostimulant compositions, such as hormones, algae extracts, and plant growth-promoting bacteria [22].

### **3. Water stress in plants**

Water availability is one of the most limiting environmental factors that affect crop productivity. In the semiarid tropics, the occurrence of drought or water deficit in the soil is quite common, despite the fact that crops in regions of tropical and temperate climate suffer seasonal periods of water deficit, especially during the summer [23].

Drought is a prevalent stress factor especially in arid and semiarid areas and can affect different aspects of plant growth, development, and metabolism. Drought is a multidimensional stress factor, and hence its effects on plants are complex. Its effects on plants can occur on a molecular level up to a whole-plant level. There are several reasons for drought in nature, including low rainfall, salinity, high temperature, and high intensity of light, among others [24].

Some of the plants' first responses to stress appear to be mediated by biophysical events, rather than changes in chemical reactions resulting from dehydration. The closing of stomata, the reduction of photosynthesis, and osmotic adjustments are the responses of some plants to the first stage of water deficit [25]. As the water content of the plant decreases, the cells shrink, and the cell walls relax. With this, the solutes increase their concentration in the cells, and the plasma membrane becomes thicker and more compressed, as it covered a smaller area than before [1]. Cell expansion occurs when the turgor pressure is greater than the growth of the cell wall. Water stress greatly decreases cell expansion and plant growth due to low turgor pressure [26].

Stomata provide the main mechanism for controlling the rate of water loss. However, the site of water loss is also the site of carbon gain by the plant, so a reduction in water loss by stomatal control also results in a reduction in assimilation with consequent effects on productivity and the accumulation of reactive oxygen species [27]. These responses hinder the supply of CO2 for photosynthesis and expose chloroplasts to excess energy excitation, especially under high light intensity [25].

The low potentials in the soil and in the plant inhibit their growth, reduce the development activities of cells and tissues, decrease the uptake of nutrients, and cause morphological and biochemical changes [28]. To maintain water uptake, the roots have to grow deeper or increase their density. A characteristic of droughtresistant species is that they have a large proportion of their total mass consisting of roots and a deep-rooted habit. A high root/shoot ratio does not indicate in itself great ability to absorb water: water deficiency invariably increases the root/shoot ratio, but this is due to the loss of plant shoot weight without loss of root mass [1].

Photosynthesis is the driving force of plant productivity. The ability to maintain the rate of photosynthetic carbon dioxide and the assimilation of nitrate under environmental stresses is fundamental for the maintenance of plant growth and production. It is known that when water stress becomes extreme, non-stomatal factors can become even more limiting for photosynthesis [17].

The water deficit often decreases the number of photons captured by the leaves because withered leaves are at a more acute angle to the sun's rays. Changes in the absorption characteristics of the leaves occur due to the shrinkage of the cells. However, changes in chloroplasts and thylakoid during light capture and energy transfer centers are relatively small under water deficit conditions [29].

### **3.1 Reactive oxygen species and water stress**

The diatomic oxygen (O2) molecules in the Earth's atmosphere are the major promoters of reactions in cells. Except for those organisms that are specially adapted to live under anaerobic conditions, all animals and plants require oxygen for efficient energy production [30].

Aerobic organisms use diatomic oxygen as a terminal electron receptor, providing a high-energy field compared to fermentation and anaerobic respiration. In this base stage, molecular oxygen is relatively nonreactive, but it is capable of giving rise to excited reactive and lethal states, such as free radicals and their derivatives [31].

Superoxide, produced by electron transport to oxygen, is not compatible with cellular metabolism; hence, all organisms that are involved in aerobic environments must have an efficient mechanism capable of removing or neutralizing free radicals from cellular components. The balance between oxidative and antioxidant capabilities determines the fate of the plant [32]. Without this defense mechanism, plants may not efficiently convert solar energy into chemical energy [33].

The formation of reactive oxygen species occurs primarily through the superoxide radical (O2 ●<sup>−</sup>), which can be dismutated into hydrogen peroxide (H2O2), or even through catalytic action, by the action of the superoxide dismutase (SOD) enzyme.

**341**

*Amelioration of Drought Stress on Plants under Biostimulant Sources*

conditions of environmental and xenobiotic stress [34].

Antioxidant systems in plants act as mechanisms of resistance to stress by protecting the membranes against damage caused by these oxygen species produced under

The fate of cells under stressful environments is determined by the duration of the stress, as well as the plant's protective capacity. Reactive oxygen species (ROS) play a crucial role in causing cellular damage to plants under stress. The sequence of events in plant tissues subjected to stress is increased production of ROS; increased levels of antioxidants; and increase in the capacity to "sweep" ROS, resulting in the

The detoxification mechanisms of ROS exist in all plants and can be categorized into enzymatic (superoxide dismutase, SOD; catalase, CAT; ascorbate peroxidase, APX, among others) and nonenzymatic (carotenoids, ascorbic acid, among others). The degree to which the amount and activities of antioxidant enzymes increases under water stress is extremely variable between many plant species and even between two cultivars of the same species. The level of response depends on the species, the development of the plant, as well as the duration and intensity of the

The superoxide produced by the thylakoid can spontaneously be dismutated into molecular oxygen and hydrogen peroxide. In chloroplasts, this reaction is catalyzed enzymatically via superoxide dismutase (SOD). Chloroplasts also contain large amounts of ascorbic acid, which can efficiently reduce superoxide to hydrogen

Plants have the superoxide dismutase enzyme containing Cu and Zn, Fe, or Mn as prosthetic metals. Zn is found in superoxide dismutase present in chloroplasts and cytosol, while Mn is found in superoxide dismutase in mitochondria and Fe in

Reactive oxygen species can react with unsaturated fatty acids, causing the peroxidation of essential lipid membranes in plasmalemma or intracellular organelles [33]. The damage caused by the peroxidation of plasmalemma leads to extravasation of cellular content and rapid dissection and cell death. The damaged intracellular membrane affects the respiratory activity in the mitochondria, in addition to

Under normal conditions, antioxidant systems eliminate or slow the reaction of reactive oxygen, preventing its transformation into products more toxic to cells. Photosynthetic cells can tolerate high levels of oxygen because endogenous mechanisms sweep and remove toxic products before cell damage occurs [32]. However, oxidative damage is evident under conditions where the rate of production of ROS

Water stress conditions can trigger an increase in the production of various forms of reactive oxygen, which can explain the damage to chloroplasts, lipids, and proteins and the alteration of the structural integrity of cell membranes. During

nonenzymatically with hydrogen peroxide (H2O2), giving rise to products such as hydroxyl radicals (OH<sup>−</sup>) and singlet oxygen (1O2), which are more reactive than the

Although a number of regulatory mechanisms have been evolved within the plant cell to limit the production of these toxic molecules, oxidative damage remains a potential problem, as it causes disturbances in metabolism, such as loss of coordination between production processes (source) and energy use (drain) during

When plants are under stress, free radicals or ROS damage plant cells, and antioxidants decrease the toxicity of these radicals. Plants with high levels of antioxidants produce better root and shoot growth, maintaining a high water content in

●<sup>−</sup>) can also react

superoxide dismutase is present in chloroplasts and mitochondria [36].

depigmentation and loss of the ability to fix carbon in chloroplasts [34].

the reduction of water inside the plant, the superoxide radical (O2

photosynthesis on green leaves under stressful environments [38].

●<sup>−</sup>) [32].

*DOI: http://dx.doi.org/10.5772/intechopen.91975*

plant's tolerance against water stress [35].

peroxide via ascorbate peroxidase [4].

is high and the removal ability is low [37].

superoxide radical (O2

stress [35].

### *Amelioration of Drought Stress on Plants under Biostimulant Sources DOI: http://dx.doi.org/10.5772/intechopen.91975*

*Plant Stress Physiology*

Some of the plants' first responses to stress appear to be mediated by biophysical events, rather than changes in chemical reactions resulting from dehydration. The closing of stomata, the reduction of photosynthesis, and osmotic adjustments are the responses of some plants to the first stage of water deficit [25]. As the water content of the plant decreases, the cells shrink, and the cell walls relax. With this, the solutes increase their concentration in the cells, and the plasma membrane becomes thicker and more compressed, as it covered a smaller area than before [1]. Cell expansion occurs when the turgor pressure is greater than the growth of the cell wall. Water stress greatly decreases cell expansion and plant growth due to low turgor pressure [26]. Stomata provide the main mechanism for controlling the rate of water loss. However, the site of water loss is also the site of carbon gain by the plant, so a reduction in water loss by stomatal control also results in a reduction in assimilation with consequent effects on productivity and the accumulation of reactive oxygen species [27]. These responses hinder the supply of CO2 for photosynthesis and expose chloroplasts to excess energy excitation, especially under high light intensity [25]. The low potentials in the soil and in the plant inhibit their growth, reduce the development activities of cells and tissues, decrease the uptake of nutrients, and cause morphological and biochemical changes [28]. To maintain water uptake, the roots have to grow deeper or increase their density. A characteristic of droughtresistant species is that they have a large proportion of their total mass consisting of roots and a deep-rooted habit. A high root/shoot ratio does not indicate in itself great ability to absorb water: water deficiency invariably increases the root/shoot ratio, but this is due to the loss of plant shoot weight without loss of root mass [1]. Photosynthesis is the driving force of plant productivity. The ability to maintain

the rate of photosynthetic carbon dioxide and the assimilation of nitrate under environmental stresses is fundamental for the maintenance of plant growth and production. It is known that when water stress becomes extreme, non-stomatal

The water deficit often decreases the number of photons captured by the leaves because withered leaves are at a more acute angle to the sun's rays. Changes in the absorption characteristics of the leaves occur due to the shrinkage of the cells. However, changes in chloroplasts and thylakoid during light capture and energy

The diatomic oxygen (O2) molecules in the Earth's atmosphere are the major promoters of reactions in cells. Except for those organisms that are specially adapted to live under anaerobic conditions, all animals and plants require oxygen

Aerobic organisms use diatomic oxygen as a terminal electron receptor, providing a high-energy field compared to fermentation and anaerobic respiration. In this base stage, molecular oxygen is relatively nonreactive, but it is capable of giving rise to excited reactive and lethal states, such as free radicals and their derivatives [31]. Superoxide, produced by electron transport to oxygen, is not compatible with cellular metabolism; hence, all organisms that are involved in aerobic environments must have an efficient mechanism capable of removing or neutralizing free radicals from cellular components. The balance between oxidative and antioxidant capabilities determines the fate of the plant [32]. Without this defense mechanism, plants

The formation of reactive oxygen species occurs primarily through the superox-

through catalytic action, by the action of the superoxide dismutase (SOD) enzyme.

●<sup>−</sup>), which can be dismutated into hydrogen peroxide (H2O2), or even

factors can become even more limiting for photosynthesis [17].

**3.1 Reactive oxygen species and water stress**

for efficient energy production [30].

transfer centers are relatively small under water deficit conditions [29].

may not efficiently convert solar energy into chemical energy [33].

**340**

ide radical (O2

Antioxidant systems in plants act as mechanisms of resistance to stress by protecting the membranes against damage caused by these oxygen species produced under conditions of environmental and xenobiotic stress [34].

The fate of cells under stressful environments is determined by the duration of the stress, as well as the plant's protective capacity. Reactive oxygen species (ROS) play a crucial role in causing cellular damage to plants under stress. The sequence of events in plant tissues subjected to stress is increased production of ROS; increased levels of antioxidants; and increase in the capacity to "sweep" ROS, resulting in the plant's tolerance against water stress [35].

The detoxification mechanisms of ROS exist in all plants and can be categorized into enzymatic (superoxide dismutase, SOD; catalase, CAT; ascorbate peroxidase, APX, among others) and nonenzymatic (carotenoids, ascorbic acid, among others). The degree to which the amount and activities of antioxidant enzymes increases under water stress is extremely variable between many plant species and even between two cultivars of the same species. The level of response depends on the species, the development of the plant, as well as the duration and intensity of the stress [35].

The superoxide produced by the thylakoid can spontaneously be dismutated into molecular oxygen and hydrogen peroxide. In chloroplasts, this reaction is catalyzed enzymatically via superoxide dismutase (SOD). Chloroplasts also contain large amounts of ascorbic acid, which can efficiently reduce superoxide to hydrogen peroxide via ascorbate peroxidase [4].

Plants have the superoxide dismutase enzyme containing Cu and Zn, Fe, or Mn as prosthetic metals. Zn is found in superoxide dismutase present in chloroplasts and cytosol, while Mn is found in superoxide dismutase in mitochondria and Fe in superoxide dismutase is present in chloroplasts and mitochondria [36].

Reactive oxygen species can react with unsaturated fatty acids, causing the peroxidation of essential lipid membranes in plasmalemma or intracellular organelles [33]. The damage caused by the peroxidation of plasmalemma leads to extravasation of cellular content and rapid dissection and cell death. The damaged intracellular membrane affects the respiratory activity in the mitochondria, in addition to depigmentation and loss of the ability to fix carbon in chloroplasts [34].

Under normal conditions, antioxidant systems eliminate or slow the reaction of reactive oxygen, preventing its transformation into products more toxic to cells. Photosynthetic cells can tolerate high levels of oxygen because endogenous mechanisms sweep and remove toxic products before cell damage occurs [32]. However, oxidative damage is evident under conditions where the rate of production of ROS is high and the removal ability is low [37].

Water stress conditions can trigger an increase in the production of various forms of reactive oxygen, which can explain the damage to chloroplasts, lipids, and proteins and the alteration of the structural integrity of cell membranes. During the reduction of water inside the plant, the superoxide radical (O2 ●<sup>−</sup>) can also react nonenzymatically with hydrogen peroxide (H2O2), giving rise to products such as hydroxyl radicals (OH<sup>−</sup>) and singlet oxygen (1O2), which are more reactive than the superoxide radical (O2 ●<sup>−</sup>) [32].

Although a number of regulatory mechanisms have been evolved within the plant cell to limit the production of these toxic molecules, oxidative damage remains a potential problem, as it causes disturbances in metabolism, such as loss of coordination between production processes (source) and energy use (drain) during photosynthesis on green leaves under stressful environments [38].

When plants are under stress, free radicals or ROS damage plant cells, and antioxidants decrease the toxicity of these radicals. Plants with high levels of antioxidants produce better root and shoot growth, maintaining a high water content in the leaves and low incidence of disease, both occurring when they are under ideal growing conditions and under environmental stress [18].

### **3.2 Biostimulants and reactive oxygen species**

The use of biostimulants in plant breeding could change the activity of enzymes and antioxidant properties. Lycopene, ascorbic acid, phenolic compounds, and others have antioxidant properties. Antioxidant compounds (e.g., phenols, ascorbic acid) and enzymes (e.g., catalase, peroxidase, superoxide dismutase) detoxify reactive oxygen molecules [20].

Biostimulants stimulate root production and growth when applied to seeds or early plant development, especially in soils with low fertility and low water availability. Biostimulants act in accelerating the recovery of the seedlings in unfavorable conditions, such as water deficit. In addition, biostimulants reduce the need of fertilizers to the plants and increase their productivity and resistance to water stress, since they act as a hormonal and nutritional increment [15].

The application of humic acid extracts seems to be beneficial for field crop monocots. In a study conducted by [39], extracts from vermicompost applied to rice (*Oryza sativa* L.) played a role in activating antioxidative enzymatic function and increased ROS-scavenging enzymes. These enzymes are required to inactivate toxicfree oxygen radicals produced in plants under drought stress. Humic acid extracts may stimulate plant growth by improving nutrient uptake by exerting hormone-like effects as auxins, stimulating shoot elongation and increasing leaf nutrient accumulation and chlorophyll biosynthesis [40].

According to [41], humic acids improve root and shoot growth by increasing the concentrations of antioxidants in tall fescue (*Festuca arundinacea*) and creeping bent grass (*Agrostis palustris*) grown under conditions of low water availability. The authors also claim that exogenous applications of seaweed extracts together with humic acids promote root and shoot growth through the action of antioxidants in plants under water stress conditions.

A study carried out using a biostimulant based on salicylic acid and chitosan nanoparticles had an effect on the enzyme and antioxidant activity in maize leaves under water shortage [42]. The enzyme activity in leaves treated with chitosan, salicylic acid, and a control was comparable, and the activity of superoxide dismutase and peroxidase activity in plants treated with a biostimulant was 7.7 (after 2 days) and 5.2 (after 3 days) times higher than for plants treated with only salicylic acid.

The activities of antioxidant enzymes in plants are normally favored when plants are subjected to some kind of improvement in the conditions in which they are grown. The superoxide dismutase (SOD) antioxidant enzyme is the first line of defense against ROS caused by environmental stresses. Increases in SOD values provide an increase in plant resistance when subjected to environmental stresses [43].

In an experiment with Kentucky bluegrass (*Poa pratensis*) subjected to water stress and humic acid applications, [44] observed an increase in superoxide dismutase activities related to the applied doses of humic acids. However, a decrease in the activity of superoxide dismutase related to soil moisture content was observed. The authors justify this decrease by the increase in nonenzymatic antioxidants favored by the action of humic acids, which caused a decrease in the reactive oxygen species present in the cells.

The activity of superoxide dismutase responds differently to water deficit in different experiments and species: it can be increased [45] or decreased [46], or it cannot be altered [45]. Due to the presence of multiple enzymatic forms of the superoxide dismutase enzyme [33], only the investigation of the responses of each

**343**

soybeans [7].

*Amelioration of Drought Stress on Plants under Biostimulant Sources*

of its enzymatic forms can provide more information about the behavior of this

Some authors mention that catalase activity has little affinity for hydrogen peroxide, a reason why it is common not to have a significant increase in its activity when evaluated in plants under stress [7]. [47] examined the activity of catalase in rice seedlings (*Oryza sativa*) under water stress and found that the increase of this enzyme in plants was not significant. Likewise, [48] did not find a significant increase for catalase in tomato plants (*Lycopersicon esculentum* Mill. cv. Nikita) submitted to three different levels of water stress. However, the extract of *Moringa oleifera* used as a biostimulant in rocket plants (*Eruca vesicaria subsp. sativa*) under water stress presented a decrease in the activity of the antioxidant enzymes (cata-

Several seaweed species influence ROS-scavenging systems in the plant tissue. Seaweed extracts controlled oxidative stress under drought conditions, by reducing lipid peroxidation, increasing total phenolic content, and enhancing superoxide dismutase, catalase, and ascorbate peroxidase activity in green bean (*Phaseolus vulgaris*) [50]. Extracts from *Sargassum* and *Ulva*, applied as seed presoaking, activated antioxidant systems by enhancing catalase and peroxidase activities, increasing ascorbic acid content, and therefore alleviating stress symptoms in wheat grown under drought conditions [51]. *Ascophyllum nodosum* extract applied to roots increased the total phenolic and flavonoid content and total antioxidant activity in spinach (*Spinacia oleracea*) [52]. In tall fescue (*Festuca arundinacea*), *A. nodosum* extract increased the activity of superoxide dismutase and in another study additionally enhanced glutathione reductase and ascorbate peroxidase activities [36]. Similarly applied seaweed extract increased the antioxidant capacity and enhanced flavonoid and tannin content in plant leaves of the ornamental hybrid *Calibrachoa* x

Seaweed extracts have also been applied in combination with other compounds to enhance antioxidant activity in plants under water stress, such as a mixture of seaweed extracts from *A. nodosum, Fucus* spp., and *Laminaria* spp. with zinc and manganese and *A. nodosum* extract with free amino acids. These mixtures increased superoxide dismutase activity in shoots and roots of maize (*Zea mays*) and soybean (*Glycine max*). Collectively, these studies demonstrate that seaweed extracts enhance antioxidant activity, indicating their potential to scavenge damaging ROS

Humic acids have also been shown to alleviate water deficit stress. Faba bean (*Vicia faba*) plants were protected from lead-induced oxidative damage by fulvic acids, which reduced lipid peroxidation, hydrogen peroxide, and pigment content [55]. The foliar application of fulvic acid ameliorated drought stress symptoms of reduced chlorophyll content, gas exchange, and yield while enhancing activities of superoxide dismutase, peroxidase, and catalase and increasing proline content in a study with maize [56]. Humic and fulvic acid based biostimulants, applied to the soil, enhanced superoxide dismutase, ascorbate peroxidase, and catalase activities in leaves of maize grown under well-watered and drought conditions. However, the effect of these biostimulants was less pronounced in

Humic substances can also increase activity of antioxidant enzymes. Activity of superoxide dismutase, peroxidase, and catalase was higher after foliar application of fulvic acid in maize grown under drought conditions. Biostimulant containing humic and fulvic acids and amino acids increased activity of antioxidant enzymes, specifically superoxide dismutase and ascorbate peroxidase in maize subjected to

*DOI: http://dx.doi.org/10.5772/intechopen.91975*

enzyme in plants subjected to water stress.

lase, peroxidase, and superoxide dismutase) [49].

*hybrida* under normal conditions [53].

molecules and improve plant stress tolerance [54].

drought stress, but did not affect catalase activity [7].

### *Amelioration of Drought Stress on Plants under Biostimulant Sources DOI: http://dx.doi.org/10.5772/intechopen.91975*

*Plant Stress Physiology*

tive oxygen molecules [20].

lation and chlorophyll biosynthesis [40].

plants under water stress conditions.

species present in the cells.

the leaves and low incidence of disease, both occurring when they are under ideal

The use of biostimulants in plant breeding could change the activity of enzymes

Biostimulants stimulate root production and growth when applied to seeds or early plant development, especially in soils with low fertility and low water availability. Biostimulants act in accelerating the recovery of the seedlings in unfavorable conditions, such as water deficit. In addition, biostimulants reduce the need of fertilizers to the plants and increase their productivity and resistance to water

The application of humic acid extracts seems to be beneficial for field crop monocots. In a study conducted by [39], extracts from vermicompost applied to rice (*Oryza sativa* L.) played a role in activating antioxidative enzymatic function and increased ROS-scavenging enzymes. These enzymes are required to inactivate toxicfree oxygen radicals produced in plants under drought stress. Humic acid extracts may stimulate plant growth by improving nutrient uptake by exerting hormone-like effects as auxins, stimulating shoot elongation and increasing leaf nutrient accumu-

According to [41], humic acids improve root and shoot growth by increasing the concentrations of antioxidants in tall fescue (*Festuca arundinacea*) and creeping bent grass (*Agrostis palustris*) grown under conditions of low water availability. The authors also claim that exogenous applications of seaweed extracts together with humic acids promote root and shoot growth through the action of antioxidants in

A study carried out using a biostimulant based on salicylic acid and chitosan nanoparticles had an effect on the enzyme and antioxidant activity in maize leaves under water shortage [42]. The enzyme activity in leaves treated with chitosan, salicylic acid, and a control was comparable, and the activity of superoxide dismutase and peroxidase activity in plants treated with a biostimulant was 7.7 (after 2 days) and 5.2 (after 3 days) times higher than for plants treated with only salicylic acid. The activities of antioxidant enzymes in plants are normally favored when plants are subjected to some kind of improvement in the conditions in which they are grown. The superoxide dismutase (SOD) antioxidant enzyme is the first line of defense against ROS caused by environmental stresses. Increases in SOD values provide an increase in plant resistance when subjected to environmental stresses [43]. In an experiment with Kentucky bluegrass (*Poa pratensis*) subjected to water stress and humic acid applications, [44] observed an increase in superoxide dismutase activities related to the applied doses of humic acids. However, a decrease in the activity of superoxide dismutase related to soil moisture content was observed. The authors justify this decrease by the increase in nonenzymatic antioxidants favored by the action of humic acids, which caused a decrease in the reactive oxygen

The activity of superoxide dismutase responds differently to water deficit in different experiments and species: it can be increased [45] or decreased [46], or it cannot be altered [45]. Due to the presence of multiple enzymatic forms of the superoxide dismutase enzyme [33], only the investigation of the responses of each

and antioxidant properties. Lycopene, ascorbic acid, phenolic compounds, and others have antioxidant properties. Antioxidant compounds (e.g., phenols, ascorbic acid) and enzymes (e.g., catalase, peroxidase, superoxide dismutase) detoxify reac-

stress, since they act as a hormonal and nutritional increment [15].

growing conditions and under environmental stress [18].

**3.2 Biostimulants and reactive oxygen species**

**342**

of its enzymatic forms can provide more information about the behavior of this enzyme in plants subjected to water stress.

Some authors mention that catalase activity has little affinity for hydrogen peroxide, a reason why it is common not to have a significant increase in its activity when evaluated in plants under stress [7]. [47] examined the activity of catalase in rice seedlings (*Oryza sativa*) under water stress and found that the increase of this enzyme in plants was not significant. Likewise, [48] did not find a significant increase for catalase in tomato plants (*Lycopersicon esculentum* Mill. cv. Nikita) submitted to three different levels of water stress. However, the extract of *Moringa oleifera* used as a biostimulant in rocket plants (*Eruca vesicaria subsp. sativa*) under water stress presented a decrease in the activity of the antioxidant enzymes (catalase, peroxidase, and superoxide dismutase) [49].

Several seaweed species influence ROS-scavenging systems in the plant tissue. Seaweed extracts controlled oxidative stress under drought conditions, by reducing lipid peroxidation, increasing total phenolic content, and enhancing superoxide dismutase, catalase, and ascorbate peroxidase activity in green bean (*Phaseolus vulgaris*) [50]. Extracts from *Sargassum* and *Ulva*, applied as seed presoaking, activated antioxidant systems by enhancing catalase and peroxidase activities, increasing ascorbic acid content, and therefore alleviating stress symptoms in wheat grown under drought conditions [51]. *Ascophyllum nodosum* extract applied to roots increased the total phenolic and flavonoid content and total antioxidant activity in spinach (*Spinacia oleracea*) [52]. In tall fescue (*Festuca arundinacea*), *A. nodosum* extract increased the activity of superoxide dismutase and in another study additionally enhanced glutathione reductase and ascorbate peroxidase activities [36]. Similarly applied seaweed extract increased the antioxidant capacity and enhanced flavonoid and tannin content in plant leaves of the ornamental hybrid *Calibrachoa* x *hybrida* under normal conditions [53].

Seaweed extracts have also been applied in combination with other compounds to enhance antioxidant activity in plants under water stress, such as a mixture of seaweed extracts from *A. nodosum, Fucus* spp., and *Laminaria* spp. with zinc and manganese and *A. nodosum* extract with free amino acids. These mixtures increased superoxide dismutase activity in shoots and roots of maize (*Zea mays*) and soybean (*Glycine max*). Collectively, these studies demonstrate that seaweed extracts enhance antioxidant activity, indicating their potential to scavenge damaging ROS molecules and improve plant stress tolerance [54].

Humic acids have also been shown to alleviate water deficit stress. Faba bean (*Vicia faba*) plants were protected from lead-induced oxidative damage by fulvic acids, which reduced lipid peroxidation, hydrogen peroxide, and pigment content [55]. The foliar application of fulvic acid ameliorated drought stress symptoms of reduced chlorophyll content, gas exchange, and yield while enhancing activities of superoxide dismutase, peroxidase, and catalase and increasing proline content in a study with maize [56]. Humic and fulvic acid based biostimulants, applied to the soil, enhanced superoxide dismutase, ascorbate peroxidase, and catalase activities in leaves of maize grown under well-watered and drought conditions. However, the effect of these biostimulants was less pronounced in soybeans [7].

Humic substances can also increase activity of antioxidant enzymes. Activity of superoxide dismutase, peroxidase, and catalase was higher after foliar application of fulvic acid in maize grown under drought conditions. Biostimulant containing humic and fulvic acids and amino acids increased activity of antioxidant enzymes, specifically superoxide dismutase and ascorbate peroxidase in maize subjected to drought stress, but did not affect catalase activity [7].

### **4. Conclusions**

The composition of biostimulants should present a variety of organic materials such as humic substances, seaweed extracts, organic matter, and amino acids in order to improve stress tolerance. The literature on biostimulants have been reporting an increase in enzyme activities involved in antioxidant functions, especially under stress conditions.

Investigations on the role of biostimulants in the physiological mode of action in plants subjected to drought stress should be continued, since considerable researches remain to be completed to gain a clearer understanding of how these products increase the physiological health of plants under water stress.

### **Author details**

Ana Carolina Feitosa de Vasconcelos Federal University of Campina Grande, Campina Grande, Brazil

\*Address all correspondence to: ana3carol@yahoo.com.br

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

**345**

*Amelioration of Drought Stress on Plants under Biostimulant Sources*

[9] Caverzan A, Passaia G, Rosa SB, Ribeiro CW, Lazzarotto F, Margis-Pinheiro M. Plant responses to stresses: Role of ascorbate peroxidase

[10] Laxa M, Liebthal M, Telman W, Chibani K, Dietz KJ. The role of the plant antioxidant system in drought tolerance. Antioxidants. 2019;**8**(4):94.

in the antioxidant protection. Genetics and Molecular Biology. 2012;**35**(4):1011-1019. DOI: 10.1590/

S1415-47572012000600016

DOI: 10.3390/antiox8040094

[11] Gajc-Wolska J, Spizewski T, Grabowska A. The effect of seaweed extracts on the yield and quality parameters of broccoli (*Brassica oleracea* var. cymosa L.) in open field production. Acta Horticulturae. 2013;**1009**:83-89

[12] Sharma HS, Fleming C, Selby C, Rao JR, Martin T. Plant biostimulants:

macroalgae and use of extracts for crop management to reduce abiotic and biotic stresses. Journal of Applied Phycology.

[13] Hamza B, Suggars A. Biostimulants: Myths and realities. Turf grass trends.

A review on the processing of

[14] O'Donnell R. W. the auxinlike effects of humic preparations from leonardite. Soil Science.

[15] Vasconcelos ACF, Chaves LHG. In: Mirmajlessi SM, editor. Biostimulants and Their Role in Improving Plant Growth under Abiotic Stresses. IntechOpen; 2019. DOI: 10.5772/intechopen.88829. Available from: https://www.intechopen. com/online-first/biostimulants-andtheir-role-in-improving-plant-growth-

2014;**26**:465-490

2001;**10**:6-10

1973;**116**(2):106-112

under-abiotic-stresses

[16] Van Oosten MJ, Pepe O, De Pascale S, Siletti S, Maggio A. The

*DOI: http://dx.doi.org/10.5772/intechopen.91975*

[1] Taiz L, Zeiger E. Plant Physiology. 3rd ed. Sunderland, Mass: Sinauer

[2] Bodner G, Nakhforoosh A, Kaul HP. Management of crop water under drought: A review. Agronomy for Sustainable Development. 2015;**35**:401- 442. DOI: 10.1007/s13593-015-0283-4

[3] Artlip TS, Wisniewski ME. Induction of proteins in response to biotic and abiotic stresses. In: Pessarakli M, editor. Handbook of Plant and Crop Physiology. 2nd ed. New York: M. Dekker; 2002.

[4] Foyer CH. The contribution of photosynthetic oxygen metabolism to oxidative stress in plants. In: Inze D, Van Montagu M, editors. Oxidative Stress in Plants. New York: Taylor & Francis;

[5] Xie X, He Z, Chen N, Tang Z, Wang Q, Cai Y. The roles of environmental factors in regulation of oxidative stress in plant. BioMed Research International. 2019;**2019**:9732325. DOI:

[6] Bulgari R, Cocetta G, Trivellini A, Vernieri P, Ferrante A. Biostimulants and crop responses: A review. Biological Agriculture and

Horticulture. 2014;**31**(1):1-17. DOI: 10.1080/01448765.2014.964649

antioxidant responses to biostimulants in maize and soybean subjected to drought. Scientia Agricola. 2009;**66**(3):395-402. DOI: 10.1590/ S0103-90162009000300015

[8] Das K, Roychoudhury A. Reactive oxygen species (ROS) and response of antioxidants as ROS-scavengers during environmental stress in plants. Frontiers in Environmental Science. 2014;**2**:53. DOI: 10.3389/fenvs.2014.00053

[7] Vasconcelos ACF, Zhang X, Ervin EH, Kiehl JC. Enzymatic

**References**

pp. 657-679

2002. pp. 33-68

10.1155/2019/9732325

Associates; 2002. p. 690

*Amelioration of Drought Stress on Plants under Biostimulant Sources DOI: http://dx.doi.org/10.5772/intechopen.91975*

### **References**

*Plant Stress Physiology*

**4. Conclusions**

under stress conditions.

**344**

**Author details**

Ana Carolina Feitosa de Vasconcelos

provided the original work is properly cited.

Federal University of Campina Grande, Campina Grande, Brazil

© 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,

The composition of biostimulants should present a variety of organic materials such as humic substances, seaweed extracts, organic matter, and amino acids in order to improve stress tolerance. The literature on biostimulants have been reporting an increase in enzyme activities involved in antioxidant functions, especially

Investigations on the role of biostimulants in the physiological mode of action

in plants subjected to drought stress should be continued, since considerable researches remain to be completed to gain a clearer understanding of how these

products increase the physiological health of plants under water stress.

\*Address all correspondence to: ana3carol@yahoo.com.br

[1] Taiz L, Zeiger E. Plant Physiology. 3rd ed. Sunderland, Mass: Sinauer Associates; 2002. p. 690

[2] Bodner G, Nakhforoosh A, Kaul HP. Management of crop water under drought: A review. Agronomy for Sustainable Development. 2015;**35**:401- 442. DOI: 10.1007/s13593-015-0283-4

[3] Artlip TS, Wisniewski ME. Induction of proteins in response to biotic and abiotic stresses. In: Pessarakli M, editor. Handbook of Plant and Crop Physiology. 2nd ed. New York: M. Dekker; 2002. pp. 657-679

[4] Foyer CH. The contribution of photosynthetic oxygen metabolism to oxidative stress in plants. In: Inze D, Van Montagu M, editors. Oxidative Stress in Plants. New York: Taylor & Francis; 2002. pp. 33-68

[5] Xie X, He Z, Chen N, Tang Z, Wang Q, Cai Y. The roles of environmental factors in regulation of oxidative stress in plant. BioMed Research International. 2019;**2019**:9732325. DOI: 10.1155/2019/9732325

[6] Bulgari R, Cocetta G, Trivellini A, Vernieri P, Ferrante A. Biostimulants and crop responses: A review. Biological Agriculture and Horticulture. 2014;**31**(1):1-17. DOI: 10.1080/01448765.2014.964649

[7] Vasconcelos ACF, Zhang X, Ervin EH, Kiehl JC. Enzymatic antioxidant responses to biostimulants in maize and soybean subjected to drought. Scientia Agricola. 2009;**66**(3):395-402. DOI: 10.1590/ S0103-90162009000300015

[8] Das K, Roychoudhury A. Reactive oxygen species (ROS) and response of antioxidants as ROS-scavengers during environmental stress in plants. Frontiers in Environmental Science. 2014;**2**:53. DOI: 10.3389/fenvs.2014.00053

[9] Caverzan A, Passaia G, Rosa SB, Ribeiro CW, Lazzarotto F, Margis-Pinheiro M. Plant responses to stresses: Role of ascorbate peroxidase in the antioxidant protection. Genetics and Molecular Biology. 2012;**35**(4):1011-1019. DOI: 10.1590/ S1415-47572012000600016

[10] Laxa M, Liebthal M, Telman W, Chibani K, Dietz KJ. The role of the plant antioxidant system in drought tolerance. Antioxidants. 2019;**8**(4):94. DOI: 10.3390/antiox8040094

[11] Gajc-Wolska J, Spizewski T, Grabowska A. The effect of seaweed extracts on the yield and quality parameters of broccoli (*Brassica oleracea* var. cymosa L.) in open field production. Acta Horticulturae. 2013;**1009**:83-89

[12] Sharma HS, Fleming C, Selby C, Rao JR, Martin T. Plant biostimulants: A review on the processing of macroalgae and use of extracts for crop management to reduce abiotic and biotic stresses. Journal of Applied Phycology. 2014;**26**:465-490

[13] Hamza B, Suggars A. Biostimulants: Myths and realities. Turf grass trends. 2001;**10**:6-10

[14] O'Donnell R. W. the auxinlike effects of humic preparations from leonardite. Soil Science. 1973;**116**(2):106-112

[15] Vasconcelos ACF, Chaves LHG. In: Mirmajlessi SM, editor. Biostimulants and Their Role in Improving Plant Growth under Abiotic Stresses. IntechOpen; 2019. DOI: 10.5772/intechopen.88829. Available from: https://www.intechopen. com/online-first/biostimulants-andtheir-role-in-improving-plant-growthunder-abiotic-stresses

[16] Van Oosten MJ, Pepe O, De Pascale S, Siletti S, Maggio A. The role of biostimulants and bioeffectors as alleviators of abiotic stress in crop plants. Chemical and Biological Technologies in Agriculture. 2017;**4**:5. DOI: 10.1186/s40538-017-0089-5

[17] Richardson AD, Aikens M, Berlyn GP, Marshall P. Drought stress and paper birch (*Betula papyrifera*) seedlings: Effects of an organic biostimulant on plant health and stress tolerance, and detection of stress effects with instrument-based, noninvasive methods. Journal of Arboriculture. 2004;**30**:52-61

[18] Su Y, Xia S, Zhong R, Wang L. Phytohormonal quantification based on biological principles. Hormone Metabolism and Signaling in Plants. 2017;**13**:431-470

[19] Nardi S, Pizzeghello D, Schiavon M, Ertani A. Plant biostimulants: Physiological responses induced by protein hydrolyzed-based products and humic substances in plant metabolism. Scientia Agricola. 2016;**73**(1):18-23. DOI: 10.1590/0103-9016-2015-0006

[20] Drobek M, Frac M, Cybulska J. Plant biostimulants: Importance of the quality and yield of horticultural crops and the improvement of plant tolerance to abiotic stress—A review. Agronomy. 2019;**9**(6):335. DOI: 10.3390/ agronomy9060335

[21] Abbas SM. The influence of biostimulants on the growth and on the biochemical composition of *Vicia faba* CV. Giza 3 beans. Romanian Biotechnological Letters. 2013;**18**(2):8061-8068

[22] Du Jardin P. The science of plant biostimulants - A bibliographic analysis, Ad hoc study report. 2012. Brussels: European Commission. Available from: http://hdl.handle.net/2268/169257 [Accessed: 20 February 2020]

[23] Levit J. Plant responses to environmental stress. New York: Academic Press; 1980. p. 486

[24] Salehi-Lisar SY, Bakhshayeshan-Agdam H. Drought stress in plants: Causes, consequences, and tolerance. In: Hossain MA, Wani SH, Bhattacharjee S, Burritt DJ, Tran LP, editors. Drought Stress Tolerance in Plants: Physiology and Biochemistry. Switzerland: Springer International Publishing; 2016. pp. 1-16. DOI: 10.1007/978-3-319-28899-4

[25] Lawlor DW. The effects of water deficit on photosynthesis. In: Smirnoff N, editor. Environment and Plant Metabolism: Flexibility and Acclimation. Oxford; Herndon: BIOS Scientific Publishers; Books International; 1995. pp. 129-160

[26] Mckersie BD, Leshem YY. Stress and Stress Coping in Cultivated Plants. Dordrecht; Boston: Kluwer Academic Publishers; 1994. p. 256

[27] Jones MM, Turner NC, Osmond CB. Mechanisms of drought resistance. In: Paleg LG, Aspinall D, editors. The Physiology and Biochemistry of Drought Resistance in Plants. Sydney; New York: Academic Press; 1981. pp. 15-38

[28] Dubey RS, Pessarakli M. Physiological mechanisms of nitrogen absorption and assimilation in plants under stressful conditions. In: Pessarakli M, editor. Handbook of Plant and Crop Physiology. New York: M. Dekker; 2002. pp. 637-655

[29] Urban L, Aarrouf J, Bidel L. Assessing the effects of water deficit on photosynthesis using parameters derived from measurements of Leaf gas exchange and of chlorophyll a fluorescence. Frontiers in Plant Science. 2017;**8**:2068. DOI: 10.3389/fpls.2017.02068

[30] Halliwell B, Gutteridge JMC. Free Radicals in Biology and Medicine. Oxford: Clarendon Press; 1989. p. 543

[31] Perl-Treves R, Perl A. Oxidative stress. In: Inze D, Van Montagu M,

**347**

*Amelioration of Drought Stress on Plants under Biostimulant Sources*

acids as an ecological pathway to protect rice plant against oxidative stress. Ecological Engineering.

[40] Baldotto MA, Baldotto LEB. Gladiolus development in response to bulb treatment with different concentrations of humic acids. Revista

[41] Zhang X, Schmidt RE. Hormonecontaining products' impact on antioxidant status of tall fescue and creeping bentgrass subjected to drought. Crop Science. 2000;**40**:1344-1349

[42] Kumaraswamy RV, Kumari S, Choudhary RC, Sharma SS, Pal A, Raliya R, et al. Salicylic acid functionalized chitosan nanoparticle: A sustainable biostimulant for plant. International Journal of Biological Macromolecules. 2019;**123**:59-69

[43] Zhang X, Ervin E, Evanylo G, Sherony C, Peot C. Biosolids impact on tall fescue drought resistance. Journal of Residuals Science and Technology.

[44] Zhang X, Schmidt RE. Antioxidant response to hormone-containing product in Kentucky bluegrass subjected to drought. Crop Science.

[45] Luna M, Badiani M, Felici M, Artemi F, Sermanni GG. Selective enzyme inactivation over water stress in maize (*Zea mays* L.) and wheat (*Triticum aestivum* L.) seedlings. Environmental and Experimental

[46] Quartacci MF, Navari-Izzo F. Water stress and free radical mediated changes in sunflower seedlings. Journal of Plant

Botany. 1985;**25**:153-156

Physiology. 1992;**139**:621-625

[47] Sharma P, Dubey RS. Drought induces oxidative stress and enhances the activities of antioxidant enzymes in

2005;**2**:173-180

1999;**39**:545-551

2012;**47**:203-208

Ceres. 2013;**60**:138-142

*DOI: http://dx.doi.org/10.5772/intechopen.91975*

editors. Oxidative Stress in Plants. New York: Taylor & Francis; 2002.

[32] Navari-Izzo F, Rascio N. Plant response to water-deficit conditions. In: Pessarakli M, editor. Handbook of Plant and Crop Stress. New York: M. Dekker;

[33] Scandalios JG. Oxygen stress and superoxide dismutase. Plant Physiology.

[34] Malan C, Greyling MM, Gressel J. Correlation between Cu/Zn superoxide dismutase and glutathione reductase, and environmental and xenobiotic stress tolerance in maize inbreeds. Plant

Science. 1990;**69**:157-166

[35] Mano J. Early events in environmental stresses in plants – Induction mechanisms of oxidative stress. In: Inze D, Van Montagu M, editors. Oxidative Stress in Plants. New York: Taylor & Francis; 2002.

[36] Zhang XZ. Influence of Plant Growth Regulators on Turfgrass Growth, Antioxidant Status, and Drought Tolerance. Blacksburg, VA: Virginia Polytechnic Institute and State

[37] Asada K, Takahashi M. Production and scavenging of active oxygen in photosynthesis. In: Kyle DJ, Osmond CB, Arntzen CJ, editors. Photoinhibition. New York: Elsevier;

[39] García AC, Santos LA, Izquierdo FG,

pp. 1-32

1999. pp. 231-270

1993;**101**:7-12

pp. 216-245

University; 1997

1987. pp. 228-287

2004;**161**:1189-1202

[38] Reddy AR, Chaitanya KV, Vivekanandan M. Drought-induced responses of photosynthesis and antioxidant metabolism in higher plants. Journal of Plant Physiology.

Sperandio MVL, Castro RN, Berbara RLL. Vermicompost humic

### *Amelioration of Drought Stress on Plants under Biostimulant Sources DOI: http://dx.doi.org/10.5772/intechopen.91975*

editors. Oxidative Stress in Plants. New York: Taylor & Francis; 2002. pp. 1-32

*Plant Stress Physiology*

role of biostimulants and bioeffectors as alleviators of abiotic stress in crop plants. Chemical and Biological Technologies in Agriculture. 2017;**4**:5. DOI: 10.1186/s40538-017-0089-5

[24] Salehi-Lisar SY, Bakhshayeshan-Agdam H. Drought stress in plants: Causes, consequences, and tolerance. In: Hossain MA, Wani SH, Bhattacharjee S, Burritt DJ, Tran LP, editors. Drought Stress Tolerance in Plants: Physiology and Biochemistry. Switzerland: Springer International Publishing; 2016. pp. 1-16.

DOI: 10.1007/978-3-319-28899-4

[25] Lawlor DW. The effects of water deficit on photosynthesis. In: Smirnoff N, editor. Environment and Plant Metabolism: Flexibility and Acclimation. Oxford; Herndon: BIOS Scientific Publishers; Books International; 1995. pp. 129-160

[26] Mckersie BD, Leshem YY. Stress and Stress Coping in Cultivated Plants. Dordrecht; Boston: Kluwer Academic

Publishers; 1994. p. 256

Press; 1981. pp. 15-38

[28] Dubey RS, Pessarakli M.

absorption and assimilation in plants under stressful conditions. In: Pessarakli M, editor. Handbook of Plant and Crop Physiology. New York:

M. Dekker; 2002. pp. 637-655

[29] Urban L, Aarrouf J, Bidel L.

DOI: 10.3389/fpls.2017.02068

Assessing the effects of water deficit on photosynthesis using parameters derived from measurements of Leaf gas exchange and of chlorophyll a fluorescence. Frontiers in Plant Science. 2017;**8**:2068.

[30] Halliwell B, Gutteridge JMC. Free Radicals in Biology and Medicine. Oxford: Clarendon Press; 1989. p. 543

[31] Perl-Treves R, Perl A. Oxidative stress. In: Inze D, Van Montagu M,

Physiological mechanisms of nitrogen

[27] Jones MM, Turner NC, Osmond CB. Mechanisms of drought resistance. In: Paleg LG, Aspinall D, editors. The Physiology and Biochemistry of Drought Resistance in Plants. Sydney; New York: Academic

Berlyn GP, Marshall P. Drought stress and paper birch (*Betula papyrifera*) seedlings: Effects of an organic biostimulant on plant health and stress tolerance, and detection of stress effects with instrument-based, noninvasive methods. Journal of Arboriculture. 2004;**30**:52-61

[17] Richardson AD, Aikens M,

[18] Su Y, Xia S, Zhong R, Wang L. Phytohormonal quantification based on biological principles. Hormone Metabolism and Signaling in Plants.

[19] Nardi S, Pizzeghello D, Schiavon M,

[20] Drobek M, Frac M, Cybulska J. Plant biostimulants: Importance of the quality and yield of horticultural crops and the improvement of plant tolerance to abiotic stress—A review. Agronomy.

Ertani A. Plant biostimulants: Physiological responses induced by protein hydrolyzed-based products and humic substances in plant metabolism. Scientia Agricola. 2016;**73**(1):18-23. DOI: 10.1590/0103-9016-2015-0006

2019;**9**(6):335. DOI: 10.3390/

[21] Abbas SM. The influence of biostimulants on the growth and on the biochemical composition of *Vicia faba* CV. Giza 3 beans. Romanian Biotechnological Letters.

[22] Du Jardin P. The science of plant biostimulants - A bibliographic analysis, Ad hoc study report. 2012. Brussels: European Commission. Available from: http://hdl.handle.net/2268/169257 [Accessed: 20 February 2020]

[23] Levit J. Plant responses to environmental stress. New York: Academic Press; 1980. p. 486

agronomy9060335

2013;**18**(2):8061-8068

2017;**13**:431-470

**346**

[32] Navari-Izzo F, Rascio N. Plant response to water-deficit conditions. In: Pessarakli M, editor. Handbook of Plant and Crop Stress. New York: M. Dekker; 1999. pp. 231-270

[33] Scandalios JG. Oxygen stress and superoxide dismutase. Plant Physiology. 1993;**101**:7-12

[34] Malan C, Greyling MM, Gressel J. Correlation between Cu/Zn superoxide dismutase and glutathione reductase, and environmental and xenobiotic stress tolerance in maize inbreeds. Plant Science. 1990;**69**:157-166

[35] Mano J. Early events in environmental stresses in plants – Induction mechanisms of oxidative stress. In: Inze D, Van Montagu M, editors. Oxidative Stress in Plants. New York: Taylor & Francis; 2002. pp. 216-245

[36] Zhang XZ. Influence of Plant Growth Regulators on Turfgrass Growth, Antioxidant Status, and Drought Tolerance. Blacksburg, VA: Virginia Polytechnic Institute and State University; 1997

[37] Asada K, Takahashi M. Production and scavenging of active oxygen in photosynthesis. In: Kyle DJ, Osmond CB, Arntzen CJ, editors. Photoinhibition. New York: Elsevier; 1987. pp. 228-287

[38] Reddy AR, Chaitanya KV, Vivekanandan M. Drought-induced responses of photosynthesis and antioxidant metabolism in higher plants. Journal of Plant Physiology. 2004;**161**:1189-1202

[39] García AC, Santos LA, Izquierdo FG, Sperandio MVL, Castro RN, Berbara RLL. Vermicompost humic

acids as an ecological pathway to protect rice plant against oxidative stress. Ecological Engineering. 2012;**47**:203-208

[40] Baldotto MA, Baldotto LEB. Gladiolus development in response to bulb treatment with different concentrations of humic acids. Revista Ceres. 2013;**60**:138-142

[41] Zhang X, Schmidt RE. Hormonecontaining products' impact on antioxidant status of tall fescue and creeping bentgrass subjected to drought. Crop Science. 2000;**40**:1344-1349

[42] Kumaraswamy RV, Kumari S, Choudhary RC, Sharma SS, Pal A, Raliya R, et al. Salicylic acid functionalized chitosan nanoparticle: A sustainable biostimulant for plant. International Journal of Biological Macromolecules. 2019;**123**:59-69

[43] Zhang X, Ervin E, Evanylo G, Sherony C, Peot C. Biosolids impact on tall fescue drought resistance. Journal of Residuals Science and Technology. 2005;**2**:173-180

[44] Zhang X, Schmidt RE. Antioxidant response to hormone-containing product in Kentucky bluegrass subjected to drought. Crop Science. 1999;**39**:545-551

[45] Luna M, Badiani M, Felici M, Artemi F, Sermanni GG. Selective enzyme inactivation over water stress in maize (*Zea mays* L.) and wheat (*Triticum aestivum* L.) seedlings. Environmental and Experimental Botany. 1985;**25**:153-156

[46] Quartacci MF, Navari-Izzo F. Water stress and free radical mediated changes in sunflower seedlings. Journal of Plant Physiology. 1992;**139**:621-625

[47] Sharma P, Dubey RS. Drought induces oxidative stress and enhances the activities of antioxidant enzymes in growing rice seedlings. Plant Growth Regulation. 2005;**46**:209-211

[48] Zgallaï H, Steppe K, Lemeur R. Effects of different levels of water stress on leaf water potential, stomatal resistance, protein and chlorophyll content and certain antioxidative enzymes in tomato plants. Journal of Integrative Plant Biology. 2006;**48**:679-685

[49] Abdalla MM. Boosting the growth of rocket plants in response to the application of *Moringa oleifera* extracts as a biostimulant. Life Sciences. 2014;**11**:1097-8135

[50] Mansori M, Chernane H, Latique S. Seaweed extract effect on water deficit and antioxidative mechanisms in bean plants (*Phaseolus vulgaris* L.). Journal of Applied Phycology. 2015;**27**(4):1689-1698

[51] Kasim WA, Hamada EAM, El-Din NGS, Eskander S. Influence of seaweed extracts on the growth, some metabolic activities and yield of wheat grown under drought stress. International Journal of Agronomy and Agricultural Research. 2015;**7**(2):173-189

[52] Fan D, Hodges DM, Zhang JZ. Commercial extract of the brown seaweed *Ascophyllum nodosum* enhances phenolic antioxidant content of spinach (*Spinacia oleracea* L.) which protects *Caenorhabditis elegans* against oxidative and thermal stress. Food Chemistry. 2015;**124**(1):195-202

[53] Elansary HO, Norrie J, Ali HM. Enhancement of Calibrachoa growth, secondary metabolites and bioactivity using seaweed extracts. BMC Complementary and Alternative Medicine. 2016;**16**:341

[54] Wozniak E, Blaszczak A, Wiatrak P, Canady M. Biostimulant mode of action. In: The Chemical Biology of Plant

Biostimulants. 1st ed. John Wiley & Sons Ltd; 2020. pp. 229-243. DOI: 10.1002/9781119357254.ch9

[55] Shahid M, Dumat C, Silvestre J, Pinelli E. Effect of fulvic acids on leadinduced oxidative stress to metal sensitive *Vicia faba* L. plant. Biol. Fertil. The Soil. 2012;**48**(6):689-697

[56] Anjum SA, Wang L, Farooq M. Fulvic acid application improves the maize performance under well-watered and drought conditions. Journal of Agronomy and Crop Science. 2011;**197**(6):409-417

**349**

**1. Introduction**

classes of humankind.

**Chapter 19**

Rats

**Abstract**

Potential Role of Plants *Hordeum* 

Hypothyroidism in Adult Female

*Hordeum vulgare* (Barley) and *Panax ginseng* have antioxidant activity referring to their diverse phytonutrient. Hypothyroidism in adult female rats was induced by pituitary-gonadal-adrenal disturbance, depleting the serum FSH levels with the elevation of corticosterone, prolactin, progesterone and testosterone hormones as well as (ERK1/2). Hypothyroidism evoked an oxidative stress status by increasing 8-hydroxy guanosine, which initiated apoptosis by uplifting apoptotic marker Caspase-3 both in serum and brain tissues. This is confirmed by the increase in the percentage of DNA-damage in the brain tissues. Significant decrease in all monoamines' levels in different brain areas, downregulation of dopamine and 5-hydroxytryptamine receptors transcription, with a significant increase in excitatory amino acids was noted. Barley and ginseng renormalized cortisol and oxidative stress markers by increasing cellular resistance to stress and potentiated the role of the immune system through phytosterol and ginsenosides, so they considered potent free radical scavengers. Barley and *Panax ginseng* ameliorate the hormonal and neural dysfunction resulting from hypothyroidism, so they are recommended for

*vulgare* L. and *Panax ginseng*

L. in Resolving the Fertility

Disorders and Stress-Induced

Oxidative Stress Arises from

*Amany Hanafy M. Elgoly and Magda H.M. Yousef*

*Lobna F. Wahman, Marwa M. Abd Rabo,* 

relieving stress and improving mood and depression.

hypothyroidism: gonadal-neural dysfunction

**Keywords:** barley, *Panax ginseng*, oxidative stress, antioxidant,

There are many evidences revealed that food intake enriched with whole grain reduces the susceptibility of the incidence of many chronic diseases. Barley (*Hordeum vulgare*) is a food source deemed to be available for all disparate social

### **Chapter 19**

*Plant Stress Physiology*

2006;**48**:679-685

2014;**11**:1097-8135

2015;**7**(2):173-189

2015;**124**(1):195-202

Medicine. 2016;**16**:341

[53] Elansary HO, Norrie J,

Ali HM. Enhancement of Calibrachoa growth, secondary metabolites and bioactivity using seaweed extracts. BMC Complementary and Alternative

[54] Wozniak E, Blaszczak A, Wiatrak P, Canady M. Biostimulant mode of action. In: The Chemical Biology of Plant

[50] Mansori M, Chernane H, Latique S. Seaweed extract effect on water deficit and antioxidative mechanisms in bean plants (*Phaseolus vulgaris* L.). Journal of Applied Phycology. 2015;**27**(4):1689-1698

[51] Kasim WA, Hamada EAM, El-Din NGS, Eskander S. Influence of seaweed extracts on the growth, some metabolic activities and yield of wheat grown under drought stress. International Journal of

Agronomy and Agricultural Research.

[52] Fan D, Hodges DM, Zhang JZ. Commercial extract of the brown seaweed *Ascophyllum nodosum* enhances phenolic antioxidant content of spinach (*Spinacia oleracea* L.) which protects *Caenorhabditis elegans* against oxidative and thermal stress. Food Chemistry.

growing rice seedlings. Plant Growth

Biostimulants. 1st ed. John Wiley & Sons Ltd; 2020. pp. 229-243. DOI: 10.1002/9781119357254.ch9

[55] Shahid M, Dumat C, Silvestre J, Pinelli E. Effect of fulvic acids on leadinduced oxidative stress to metal sensitive *Vicia faba* L. plant. Biol. Fertil.

[56] Anjum SA, Wang L, Farooq M. Fulvic acid application improves the maize performance under well-watered and drought conditions. Journal of Agronomy and Crop Science.

The Soil. 2012;**48**(6):689-697

2011;**197**(6):409-417

[48] Zgallaï H, Steppe K, Lemeur R. Effects of different levels of water stress on leaf water potential, stomatal resistance, protein and chlorophyll content and certain antioxidative enzymes in tomato plants. Journal of Integrative Plant Biology.

[49] Abdalla MM. Boosting the growth of rocket plants in response to the application of *Moringa oleifera* extracts as a biostimulant. Life Sciences.

Regulation. 2005;**46**:209-211

**348**

Potential Role of Plants *Hordeum vulgare* L. and *Panax ginseng* L. in Resolving the Fertility Disorders and Stress-Induced Oxidative Stress Arises from Hypothyroidism in Adult Female Rats

*Lobna F. Wahman, Marwa M. Abd Rabo, Amany Hanafy M. Elgoly and Magda H.M. Yousef*

### **Abstract**

*Hordeum vulgare* (Barley) and *Panax ginseng* have antioxidant activity referring to their diverse phytonutrient. Hypothyroidism in adult female rats was induced by pituitary-gonadal-adrenal disturbance, depleting the serum FSH levels with the elevation of corticosterone, prolactin, progesterone and testosterone hormones as well as (ERK1/2). Hypothyroidism evoked an oxidative stress status by increasing 8-hydroxy guanosine, which initiated apoptosis by uplifting apoptotic marker Caspase-3 both in serum and brain tissues. This is confirmed by the increase in the percentage of DNA-damage in the brain tissues. Significant decrease in all monoamines' levels in different brain areas, downregulation of dopamine and 5-hydroxytryptamine receptors transcription, with a significant increase in excitatory amino acids was noted. Barley and ginseng renormalized cortisol and oxidative stress markers by increasing cellular resistance to stress and potentiated the role of the immune system through phytosterol and ginsenosides, so they considered potent free radical scavengers. Barley and *Panax ginseng* ameliorate the hormonal and neural dysfunction resulting from hypothyroidism, so they are recommended for relieving stress and improving mood and depression.

**Keywords:** barley, *Panax ginseng*, oxidative stress, antioxidant, hypothyroidism: gonadal-neural dysfunction

### **1. Introduction**

There are many evidences revealed that food intake enriched with whole grain reduces the susceptibility of the incidence of many chronic diseases. Barley (*Hordeum vulgare*) is a food source deemed to be available for all disparate social classes of humankind.

In the Arab culture, *Hordeum vulgare* or barley syrup is used to relieve depression. It is categorized into the spring and winter types, which are considered tworowed or six-rowed depending on the number of seed rows on each spike. Based on its grain composition, barley is further classified into normal, waxy or high amylose starch type [1].

Barley has found to be enriched with valuable minerals (iron, selenium, potassium, calcium, phosphorous; zinc), phytoestrol (β-sitosterol, campesterol, stigmasterol), polyphenol (ferulic, p-coumaric, sinapic, vanillic and p-hydroxybenzoic acids, cinnamic acid derivatives, proanthocyanidins, quinines, flavonols, chalcones, flavones, flavanones, and amino phenolic compound), water-soluble vitamins (C; B1; B2; folic acid and B12), β-glucan, dietary soluble fiber, vitamin E; nicotinic acid; pyridoxine; folic acid; essential amino acids, such as tryptophan and phenylalanine; neutral amino acids (LNAA), such as the three branched-chain aromatic amino acids leucine, isoleucine, and valine [2, 3]. So, barley grain exhibits potential antioxidant and antiproliferative actions because its powerful phytochemical compounds that have been shown to lower the risk of many diseases [4, 5]. The diverse phytonutrient of barley implicates its protection activity against certain types of cancers, cardiovascular disease, arthritis, diabetic, and hypercholesterolemia. It also increases cellular energy to sustain the body homeostasis [6, 7] and modulating endocrine and neurotransmitters functions [8].

Red ginseng represents an important position as a health functional food**.** It belongs to the Panax genus of the *Araliaceae* family, ginseng characterized by a complex activity profile that includes antioxidant, anti-inflammatory, antiapoptotic, and immune-stimulatory properties and has the effects of stabilizing and balancing the entire physiology [9]. So, in Asian countries; Korea, China, and Japan, ginseng used as a therapeutic agent for a variety of diseases [10].

The major active ingredients of *Panax ginseng* are saponins, which are triterpene glycosides called "ginsenosides". Other active components include proteins, peptides, and alkaloids, which are nitrogenous compounds; polyacetylene, which is a fat-soluble component; polysaccharides and other flavonoids; fatty acids, organic acids, vitamins, sugars, inorganic salts, sterols, oligopeptides [11, 12]. Ginsenosides can be classified into three categories: the panaxadiol group (e.g. Rb1, Rb2, Rb3, Rc, Rd., Rg3, Rh2, Rs1), the panaxatriol group (e.g. Re, Rf, Rg1, Rg2, Rh1), and the oleanolic acid group (Ro) [13, 14].

Ginsenosides are lipophilic compounds so they can pass easily through the cell membrane by simple diffusion and bind to its intracellular target proteins in the cytoplasm and nucleus. Ginseng also contains more than 10 phenolic compounds that possess antioxidant biological properties that have ability to lower the effect of oxidative stress [15]. Phytoestrogens, such as genistein is an important component of ginseng, have shown protective effects on conditions related to decreased estrogen, including menopause, osteoporosis, and cognitive disorders [13, 16].

Thyroid-stimulating hormone (TSH) is synthesized and secreted by the adenohypophysis lobe and exerts its effect by binding to the cognate thyrotropin receptor (TSHR) to stimulates the production of thyroglobulin and thyroid peroxidase proteins, which are essential for the synthesis and secretion of thyroid hormones (THs) [17]. THs bind their nuclear receptors (TRs), which are present in many tissues and organs in the human body and hence regulate their functions [18].

Thyroid hormones influence a wide range of brain developmental processes, such as myelination, neuronal and glial cell differentiation by regulating the gene involved in these processes thus hypothyroidism may reduce axonal growth and dendritic arborization in the cerebral cortex, visual cortex, auditory cortex, hippocampus, and cerebellum, as well as impaired memory, cognitive function and attentiveness [3, 19].

**351**

**Table 1.**

*Nutrition facts in barley/100 g.*

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility…*

THs plays a role in neurotransmitter release from their storage vesicles such as norepinephrine (NE), epinephrine (E), serotonin (5-HT) and dopamine (DA) and hence maintaining good mental state, mood regulation, modulating post-receptor signal transduction, gene expression and preventing depression [19]. So, hypothyroidism is a highly prevalent condition that impairs learning, memory, induce delayed skeletal development, cardiovascular diseases, secondary hypertension, the deterioration of

The study was carried out by using adult female Wistar albino rats weighing 180–200 g. Animals were housed at 23 ± 2°C and 55 ± 5% humidity with a 12 h light/dark cycle rats were provided a standard diet and water *ad libitum.*

Barley was prepared as an emulsion in water (1 g ground barley soaked in 10 ml of distilled water) and administered daily *per* [21]. The nutritional facts of barley

Dried roots of the Korean *Panax ginseng* were obtained as a brown powder and dissolved in distilled water. Animals received a daily oral dose of 1.8 mg/200 g body

Neo-Mercazole is the least toxic anti-thyroid agent within therapeutic dose ranges [23] therefore it was selected for hypothyroidism induction. The animals were orally administered a daily dose of 5.0 mg.kg—of Neo-Mercazole for 1 month [24]. Hypothyroidism was manifested by the increased level of serum TSH associ-

Carbohydrates (78.2 g) Vitamin B6 (0.29 μg) Choline (38 mg) Fibers (15.5 g) Vitamin K (2.5 μg) Riboflavin (B2) (0.124 μg) Energy (350 kcal) Niacin (B3) (4.8 μg) Calcium (30 mg) Fat (1.2 g) Pantothenic acid (B5) (0.29 μg) Iron (3.5 mg) Protein (10 g) Thiamine (B1) (0.2 μg) Magnesium (80 mg) Vitamin A (15 μg) Folic acid (25 μg) Phosphorus (200 mg)

*DOI: http://dx.doi.org/10.5772/intechopen.92359*

human reproductive health and brain dysfunction [20].

**2. Materials and methods used in the research**

**2.2 Preparation of** *Hordeum vulgare* **(barley)**

weight (equivalent to the therapeutic dose [22]) for 30 days.

Zinc (2.5 mg) Potassium (250 mg)

per 100 g are presented in **Table 1**.

**2.3 Preparation of** *Panax ginseng*

**2.4 Induction of hypothyroidism**

ated with low level of f T4.

**2.1 Animals**

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility… DOI: http://dx.doi.org/10.5772/intechopen.92359*

THs plays a role in neurotransmitter release from their storage vesicles such as norepinephrine (NE), epinephrine (E), serotonin (5-HT) and dopamine (DA) and hence maintaining good mental state, mood regulation, modulating post-receptor signal transduction, gene expression and preventing depression [19]. So, hypothyroidism is a highly prevalent condition that impairs learning, memory, induce delayed skeletal development, cardiovascular diseases, secondary hypertension, the deterioration of human reproductive health and brain dysfunction [20].

### **2. Materials and methods used in the research**

### **2.1 Animals**

*Plant Stress Physiology*

starch type [1].

endocrine and neurotransmitters functions [8].

oleanolic acid group (Ro) [13, 14].

In the Arab culture, *Hordeum vulgare* or barley syrup is used to relieve depression. It is categorized into the spring and winter types, which are considered tworowed or six-rowed depending on the number of seed rows on each spike. Based on its grain composition, barley is further classified into normal, waxy or high amylose

Barley has found to be enriched with valuable minerals (iron, selenium, potassium, calcium, phosphorous; zinc), phytoestrol (β-sitosterol, campesterol, stigmasterol), polyphenol (ferulic, p-coumaric, sinapic, vanillic and p-hydroxybenzoic acids, cinnamic acid derivatives, proanthocyanidins, quinines, flavonols, chalcones, flavones, flavanones, and amino phenolic compound), water-soluble vitamins (C; B1; B2; folic acid and B12), β-glucan, dietary soluble fiber, vitamin E; nicotinic acid; pyridoxine; folic acid; essential amino acids, such as tryptophan and phenylalanine; neutral amino acids (LNAA), such as the three branched-chain aromatic amino acids leucine, isoleucine, and valine [2, 3]. So, barley grain exhibits potential antioxidant and antiproliferative actions because its powerful phytochemical compounds that have been shown to lower the risk of many diseases [4, 5]. The diverse phytonutrient of barley implicates its protection activity against certain types of cancers, cardiovascular disease, arthritis, diabetic, and hypercholesterolemia. It also increases cellular energy to sustain the body homeostasis [6, 7] and modulating

Red ginseng represents an important position as a health functional food**.** It belongs to the Panax genus of the *Araliaceae* family, ginseng characterized by a complex activity profile that includes antioxidant, anti-inflammatory, antiapoptotic, and immune-stimulatory properties and has the effects of stabilizing and balancing the entire physiology [9]. So, in Asian countries; Korea, China, and

The major active ingredients of *Panax ginseng* are saponins, which are triterpene glycosides called "ginsenosides". Other active components include proteins, peptides, and alkaloids, which are nitrogenous compounds; polyacetylene, which is a fat-soluble component; polysaccharides and other flavonoids; fatty acids, organic acids, vitamins, sugars, inorganic salts, sterols, oligopeptides [11, 12]. Ginsenosides can be classified into three categories: the panaxadiol group (e.g. Rb1, Rb2, Rb3, Rc, Rd., Rg3, Rh2, Rs1), the panaxatriol group (e.g. Re, Rf, Rg1, Rg2, Rh1), and the

Ginsenosides are lipophilic compounds so they can pass easily through the cell membrane by simple diffusion and bind to its intracellular target proteins in the cytoplasm and nucleus. Ginseng also contains more than 10 phenolic compounds that possess antioxidant biological properties that have ability to lower the effect of oxidative stress [15]. Phytoestrogens, such as genistein is an important component of ginseng, have shown protective effects on conditions related to decreased estrogen,

Thyroid-stimulating hormone (TSH) is synthesized and secreted by the adenohypophysis lobe and exerts its effect by binding to the cognate thyrotropin receptor (TSHR) to stimulates the production of thyroglobulin and thyroid peroxidase proteins, which are essential for the synthesis and secretion of thyroid hormones (THs) [17]. THs bind their nuclear receptors (TRs), which are present in many tissues and

Thyroid hormones influence a wide range of brain developmental processes, such as myelination, neuronal and glial cell differentiation by regulating the gene involved in these processes thus hypothyroidism may reduce axonal growth and dendritic arborization in the cerebral cortex, visual cortex, auditory cortex, hippocampus, and cerebellum, as well as impaired memory, cognitive function and attentiveness [3, 19].

Japan, ginseng used as a therapeutic agent for a variety of diseases [10].

including menopause, osteoporosis, and cognitive disorders [13, 16].

organs in the human body and hence regulate their functions [18].

**350**

The study was carried out by using adult female Wistar albino rats weighing 180–200 g. Animals were housed at 23 ± 2°C and 55 ± 5% humidity with a 12 h light/dark cycle rats were provided a standard diet and water *ad libitum.*

### **2.2 Preparation of** *Hordeum vulgare* **(barley)**

Barley was prepared as an emulsion in water (1 g ground barley soaked in 10 ml of distilled water) and administered daily *per* [21]. The nutritional facts of barley per 100 g are presented in **Table 1**.

### **2.3 Preparation of** *Panax ginseng*

Dried roots of the Korean *Panax ginseng* were obtained as a brown powder and dissolved in distilled water. Animals received a daily oral dose of 1.8 mg/200 g body weight (equivalent to the therapeutic dose [22]) for 30 days.

### **2.4 Induction of hypothyroidism**

Neo-Mercazole is the least toxic anti-thyroid agent within therapeutic dose ranges [23] therefore it was selected for hypothyroidism induction. The animals were orally administered a daily dose of 5.0 mg.kg—of Neo-Mercazole for 1 month [24]. Hypothyroidism was manifested by the increased level of serum TSH associated with low level of f T4.


### **Table 1.** *Nutrition facts in barley/100 g.*

### **2.5 Experimental design of barley work**

Animals randomly divided into equally four-treatment groups. Except for euthyroid animals (EU) (groups 1&2), hypothyroid animals (H) (group 3&4) were orally administered 5.0 mg kg<sup>−</sup><sup>1</sup> bwt Neo-Mercazole until the end of the study. Following 30 days of Neo-Mercazole administration, groups 2 and 4 orally administered 100 mg kg<sup>−</sup><sup>1</sup> bwt barley (B) [21] water suspension for 4 weeks. The four groups named: EU; EU + B; H; H + B.

### **2.6 Experimental design of** *Panax ginseng* **work**

Rats were divided equally into four groups. First group was an intact control group that received distilled water. The second group was the hypothyroid group (H group) which orally treated with 5 mg kg<sup>−</sup><sup>1</sup> body- weight Neo-Mercazole for 30 days for induction of hypothyroidism. The third group was orally administered *Panax ginseng* (G group) in a daily oral dose of 1.8 mg/200 g body weight for 30 days. The fourth group receiving both Neo-Mercazole for 30 days and followed by *Panax ginseng* (H + G group) for another 30 days.

### **2.7 Blood and tissue collection**

At the end of treatment, the animals were anesthetized with 1% isoflurane followed by decapitation [25], blood was collected into serum preparation tube and the separated serum was collected and divided into aliquots, stored at −20°C for further hormones assay. The whole brains were removed from 10 rats from each group and the hypothalamus, hippocampus, cerebral cortex, midbrain, and cerebellum were dissected using a sharp blade. From another 10 rats' whole brain and thyroid gland were immediately removed and stored in ice-cold saline at −20°C for further biochemical and comet assay.

### **2.8 Methods**

Levels of f T3, f T4, ERK1/2, 8-hydroxy-2′-de-oxyguanosine (8-OhdG) and apoptotic marker Caspase-3 were determined using ELISA kit specific for rats according to manufacturer's instruction (Glory Science Co., Ltd., USA). Serum corticosteroid and gonadal hormones were determined using the ELISA kit according to the instruction of BioCheck (BioCheck Co., Ltd., USA). DNA degradation in brain and thyroid homogenates was determined by using the Comet technique according to the method [26]. Determination of monoamines in brain areas were carried according to methods of [19, 27] while free amino acids were done according to the method of [28] using the precolumn phenylisothiocyanate (PTC) derivatization technique.

### **3. The antioxidant effect of barley on fertility disorders and oxidative stress induced by hypothyroidism**

The study was conducted to address the potential ameliorative effect of barley on the disturbance in adrenal pituitary-gonadal hormones, as well as oxidative stress following hypothyroid induction. Hypothyroidism induction caused disturbances in adrenal, pituitary and gonadal hormones (**Figures 1**–**3**). Barley reversed the effect of the antithyroid drug on the levels of thyroid hormones (TSH, f T4) and their transporting proteins (TBG, TTR) as shown in **Table 2** due to its higher iron (Fe) content which plays a crucial role in modulating thyroid peroxidase

**353**

**Figure 2.**

*treated group (HT).*

**Figure 1.**

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility…*

*(A) Serum TSH mIU/L, (B) serum fT4 μg/ml, and (C) serum f T3 in control (CO) group, hypothyroid* 

*Serum corticosterone (ug/dl) (A), serum prolactin (ng/ml) (B), serum FSH (mIU/ml) (C) and serum LH (ng/l) (D) in control (CO) group, hypothyroid (H) group, barley-treated (T) group and hypothyroid-barley-*

*(H) group, barley treated (T) group and hypothyroid-barley-treated group (HT).*

*DOI: http://dx.doi.org/10.5772/intechopen.92359*

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility… DOI: http://dx.doi.org/10.5772/intechopen.92359*

**Figure 1.**

*Plant Stress Physiology*

istered 100 mg kg<sup>−</sup><sup>1</sup>

**2.5 Experimental design of barley work**

were orally administered 5.0 mg kg<sup>−</sup><sup>1</sup>

groups named: EU; EU + B; H; H + B.

**2.6 Experimental design of** *Panax ginseng* **work**

group) which orally treated with 5 mg kg<sup>−</sup><sup>1</sup>

*ginseng* (H + G group) for another 30 days.

for further biochemical and comet assay.

**stress induced by hypothyroidism**

**2.8 Methods**

**2.7 Blood and tissue collection**

Animals randomly divided into equally four-treatment groups. Except for euthyroid animals (EU) (groups 1&2), hypothyroid animals (H) (group 3&4)

Following 30 days of Neo-Mercazole administration, groups 2 and 4 orally admin-

Rats were divided equally into four groups. First group was an intact control group that received distilled water. The second group was the hypothyroid group (H

for induction of hypothyroidism. The third group was orally administered *Panax ginseng* (G group) in a daily oral dose of 1.8 mg/200 g body weight for 30 days. The fourth group receiving both Neo-Mercazole for 30 days and followed by *Panax* 

At the end of treatment, the animals were anesthetized with 1% isoflurane followed by decapitation [25], blood was collected into serum preparation tube and the separated serum was collected and divided into aliquots, stored at −20°C for further hormones assay. The whole brains were removed from 10 rats from each group and the hypothalamus, hippocampus, cerebral cortex, midbrain, and cerebellum were dissected using a sharp blade. From another 10 rats' whole brain and thyroid gland were immediately removed and stored in ice-cold saline at −20°C

Levels of f T3, f T4, ERK1/2, 8-hydroxy-2′-de-oxyguanosine (8-OhdG) and apoptotic marker Caspase-3 were determined using ELISA kit specific for rats according to manufacturer's instruction (Glory Science Co., Ltd., USA). Serum corticosteroid and gonadal hormones were determined using the ELISA kit according to the instruction of BioCheck (BioCheck Co., Ltd., USA). DNA degradation in brain and thyroid homogenates was determined by using the Comet technique according to the method [26]. Determination of monoamines in brain areas were carried according to methods of [19, 27] while free amino acids were done according to the method of [28] using the precolumn phenylisothiocyanate (PTC) derivatization technique.

**3. The antioxidant effect of barley on fertility disorders and oxidative** 

The study was conducted to address the potential ameliorative effect of barley on the disturbance in adrenal pituitary-gonadal hormones, as well as oxidative stress following hypothyroid induction. Hypothyroidism induction caused disturbances in adrenal, pituitary and gonadal hormones (**Figures 1**–**3**). Barley reversed the effect of the antithyroid drug on the levels of thyroid hormones (TSH, f T4) and their transporting proteins (TBG, TTR) as shown in **Table 2** due to its higher iron (Fe) content which plays a crucial role in modulating thyroid peroxidase

bwt barley (B) [21] water suspension for 4 weeks. The four

bwt Neo-Mercazole until the end of the study.

body- weight Neo-Mercazole for 30 days

**352**

*(A) Serum TSH mIU/L, (B) serum fT4 μg/ml, and (C) serum f T3 in control (CO) group, hypothyroid (H) group, barley treated (T) group and hypothyroid-barley-treated group (HT).*

### **Figure 2.**

*Serum corticosterone (ug/dl) (A), serum prolactin (ng/ml) (B), serum FSH (mIU/ml) (C) and serum LH (ng/l) (D) in control (CO) group, hypothyroid (H) group, barley-treated (T) group and hypothyroid-barleytreated group (HT).*

### **Figure 3.**

*(A) Serum estradiol (pmol/L), (B) serum P (ng/ml) and (C) serum T (ng/ml), in control group (CO), hypothyroid group (H), barley-treated group (T) and hypothyroid-barley-treated group (HT).*


*All data in tables represented by mean ± SD, n = 10 animals.*

*\*p < 0.05, \*\*p < 0.01 and \*\*\*p < 0.001.*

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

### **Table 2.**

*Effect of barley on THs, TTR and TBG in brain tissue of EU- and H-groups.*

(TPO) enzyme activity [2, 29]. Oxidative stress is related to hormonal disorders in a reciprocal way so in our study, the hyper TSH level stimulated the synthesis of corticosterone, and generated a state of oxidative stress which inhibited the pituitary gonadotropin [30, 31], and cause FSH depletion with the non-significant decrease of LH levels in the hypothyroid group. Also, lower levels of estradiol in the hypothyroid group associated with high progesterone and prolactin levels could be attributed to high ERK1/2 level (**Figure 4**). Barley, with its high content of phytosterol, could modulate ER-α, and β expression, augmented estradiol levels, in turn, led to activate negative feedback mechanism of pituitary-gonadal adrenal axis function and renormalize the disturbances of endocrine gland elicited by hypothyroidism.

Oxidative stress (ROS) is an imbalance between the production of pro-oxidant substances and antioxidant defense. Hypothyroidism augments the oxidative

**355**

**Figure 5.**

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility…*

insult, impairing the brain by increasing nitric oxide (NO) and NO synthase (NOS) levels in the hippocampus, which affects the lipid composition of rat brain tissues and induces DNA damage [32, 33]. In present work, hypothyroidism was associated with the high significant increase in 8-hydroxyguanosine (an oxidative stress marker), together with marked elevation in Caspase-3 (an apoptotic marker) in serum and brain tissue (**Figures 5** and **6**). These findings were confirmed by alkaline comet assays of thyroid and brain tissue homogenates. As DNA was degraded, it converted from a supercoiled form to a comet-like shape with a measurable tail length so our study revealed that the hypothyroid status induced a significant increase in the tail length, in the thyroid and brain tissues. Treatment with barley attenuated the oxidative stress status induced by hypothyroid status; it significantly decreased 8-OH guanosine levels and Caspase-3 activity. This antioxidant activity of barley could be attributed to flavonoids, ferulic, sinapic and ß- hydroxy acids (BHA) content, the major predominant polyphenol, in barley with their potent free radical scavengers by absorbing and neutralizing oxygen radicals [21, 34, 35]. Also, DNA damage was repaired by the antioxidant activity of barley as illustrated in **Figures 7** and **8**, which refer to vitamins A and E that essential for nucleotide, DNA biosynthesis, DNA repair, and methylation [1, 2, 19, 36]. Additionally, zinc protects against oxidative stress by stabilizing membranes through the inhibition of the enzyme nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase) and the stimulating of the synthesis of metallothioneins, which reduce the levels of hydroxyl radicals and sequester ROS produced in response to hypothyroidism.

*Serum 8-hydroxy guanosine (8-OHG) (ng/L) (A), Caspase-3 (ng/ml) (B) in control group (CO), hypothyroid* 

*group (H), barley-treated group (T) and hypothyroid-barley-treated group (HT).*

*(A) Serum E2/T ratio (B) serum ERK1/2 (pg/ml) in control group (CO), group hypothyroid (H),* 

*barley-treated group (T) and hypothyroid-barley-treated group (HT).*

*DOI: http://dx.doi.org/10.5772/intechopen.92359*

**Figure 4.**

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility… DOI: http://dx.doi.org/10.5772/intechopen.92359*

**Figure 4.**

*Plant Stress Physiology*

**354**

**Table 2.**

**Figure 3.**

(TPO) enzyme activity [2, 29]. Oxidative stress is related to hormonal disorders in a reciprocal way so in our study, the hyper TSH level stimulated the synthesis of corticosterone, and generated a state of oxidative stress which inhibited the pituitary gonadotropin [30, 31], and cause FSH depletion with the non-significant decrease of LH levels in the hypothyroid group. Also, lower levels of estradiol in the hypothyroid group associated with high progesterone and prolactin levels could be attributed to high ERK1/2 level (**Figure 4**). Barley, with its high content of phytosterol, could modulate ER-α, and β expression, augmented estradiol levels, in turn, led to activate negative feedback mechanism of pituitary-gonadal adrenal axis function and renormalize the disturbances of endocrine gland elicited by hypothyroidism. Oxidative stress (ROS) is an imbalance between the production of pro-oxidant

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

*All data in tables represented by mean ± SD, n = 10 animals.*

*Effect of barley on THs, TTR and TBG in brain tissue of EU- and H-groups.*

*\*p < 0.05, \*\*p < 0.01 and \*\*\*p < 0.001.*

f T4 (μg/ml) 4.43 ± 0.11 4.21 ± 0.14 3.09 ± 0.25 (a\*) 4.46 ± 0.19(b\*\*) TSH (mIU/ml) 12.47 ± 0.42 12.73 ± 0.62 15.97 ± 0.26 (a\*) 12.53 ± 0.29 (b\*) TTR (ng/ml) 51.23 ± 2.39 45.64 ± 6.53 (a\*) 16.85 ± 1.88 (a\*\*\*) 34.25 ± 1.65 (a\*\* b\*\*\*) TBG (pg/ml) 1.58 ± 0.04 2.35 ± 0.16 (a\*\*) 2.09 ± 0.04 (a\*) 2.57 ± 0.14 (a\*\*\*b\*) ERK1/2 (pg/ml) 43.67 ± 1.53 46.33 ± 1.53(a\*) 33.19 ± 2.16 (a\*\*) 47.94 ± 0.60(b\*\*)

*(A) Serum estradiol (pmol/L), (B) serum P (ng/ml) and (C) serum T (ng/ml), in control group (CO), hypothyroid group (H), barley-treated group (T) and hypothyroid-barley-treated group (HT).*

**EU EU +B H H+B**

substances and antioxidant defense. Hypothyroidism augments the oxidative

*(A) Serum E2/T ratio (B) serum ERK1/2 (pg/ml) in control group (CO), group hypothyroid (H), barley-treated group (T) and hypothyroid-barley-treated group (HT).*

insult, impairing the brain by increasing nitric oxide (NO) and NO synthase (NOS) levels in the hippocampus, which affects the lipid composition of rat brain tissues and induces DNA damage [32, 33]. In present work, hypothyroidism was associated with the high significant increase in 8-hydroxyguanosine (an oxidative stress marker), together with marked elevation in Caspase-3 (an apoptotic marker) in serum and brain tissue (**Figures 5** and **6**). These findings were confirmed by alkaline comet assays of thyroid and brain tissue homogenates. As DNA was degraded, it converted from a supercoiled form to a comet-like shape with a measurable tail length so our study revealed that the hypothyroid status induced a significant increase in the tail length, in the thyroid and brain tissues. Treatment with barley attenuated the oxidative stress status induced by hypothyroid status; it significantly decreased 8-OH guanosine levels and Caspase-3 activity. This antioxidant activity of barley could be attributed to flavonoids, ferulic, sinapic and ß- hydroxy acids (BHA) content, the major predominant polyphenol, in barley with their potent free radical scavengers by absorbing and neutralizing oxygen radicals [21, 34, 35]. Also, DNA damage was repaired by the antioxidant activity of barley as illustrated in **Figures 7** and **8**, which refer to vitamins A and E that essential for nucleotide, DNA biosynthesis, DNA repair, and methylation [1, 2, 19, 36]. Additionally, zinc protects against oxidative stress by stabilizing membranes through the inhibition of the enzyme nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase) and the stimulating of the synthesis of metallothioneins, which reduce the levels of hydroxyl radicals and sequester ROS produced in response to hypothyroidism.

**Figure 5.**

*Serum 8-hydroxy guanosine (8-OHG) (ng/L) (A), Caspase-3 (ng/ml) (B) in control group (CO), hypothyroid group (H), barley-treated group (T) and hypothyroid-barley-treated group (HT).*

**Figure 6.** *Effect of barley on caspase-3 (ng/ml) and 8-OhdG (ng/ml)in brain tissue.*

**Figure 7.**

*Effect of barley on DNA damage in thyroid tissue (A) fluorescence photomicrograph showing comets in EU-, H-, EU + B and H + B-groups. The indicated the intact DNA and indicated the degree of damaged DNA (B) tail length expressed in μm in thyroid tissue of all treated groups.*

### **Figure 8.**

*Effect of barley on DNA damage in whole brain tissue (A) fluorescence photomicrograph showing comets in EU-, H-, EU + B and H + B-groups. The indicated the intact DNA and indicated the degree of damaged DNA (B) tail length expressed in μm in whole brain tissue of all treated groups.*

**357**

**Table 3.**

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility…*

Neurotransmitters NE, DA, and 5-HT levels were significantly reduced in all brain areas (cerebellum, midbrain, cerebral cortex, hypothalamus, and hippocampus) with hypothyroidism induction as shown in **Table 3**, these could be attributed to the reduced oestradiol level as mentioned above [37, 38]. The administration of *Hordeum vulgare* (barley) improved the disturbances in the dopaminergic, serotonergic and noradrenergic pathways via two different mechanisms; first, the high phytosterol content modulates estrogen receptors (Erα and β) expression and elevate oestradiol levels. Second, barley is enriched with tryptophan and phenylalanine and could regulate the synthesis of 5-HT, DA and NE through the conversion of tryptophan to 5-hydroxytryptophan (5-HTP) to 5-HT and hydrolysis of phenylalanine to generate tyrosine that ultimately produces DA and NE [2, 36, 39, 40]. In the present study, hypothyroidism induced a significant increase in inhibitory amino acid, including GABA and histidine, which is an

These results could explain the increase in Caspase-3, which may be attributed to reduced blood oxygen–glucose levels in several brain regions as a result of increased GABA levels. The study also revealed an increase in dopamine receptors, whereas serotonin receptors were significantly decreased. The *Hordeum vulgare* (barley) treatment in the present study caused a renormalization the observed disturbances in the amino acid and neurotransmitter levels because it is enriched with folic acid, which is involved in the synthesis of monoamine neurotransmitters and modulate serotonergic, dopaminergic and noradrenergic systems by acting as a cofactor for enzymes that convert tryptophan to 5-HT and enzymes that convert tyrosine to noradrenaline [19]. The alteration in serotonin receptor densities was restored by the barley administration, due to its enriched levels of tryptophan, which is metabolized to serotonin [36, 40] and activates these receptors (**Figure 11**).

**Frontal cortex Hippocampus Hypothalamus Mid brain Cerebellum**

EU 0.52 ± 0.03 0.69 ± 0.02 0.40 ± 0.01 0.70 ± 0.01 0.58 ± 0.01 EU + B 0.45 ± 0.02 0.65 ± 0.01 0.40 ± 0.01 0.69 ± 0.01 0.60 ± 0.01 H 0.21 ± 0.09a\*\*\* 0.31 ± 0.01a\*\*\* 0.13 ± 0.01a\*\*\* 0.35 ± 0.01a\*\*\* 0.29 ± 0.01a\*\* H + B 0.35 ± 0.01a\*\*b\* 0.46 ± 0.01a\*\*b\*\* 0.22 ± 0.01a\*\*b\*\* 0.48 ± 0.01a\*\*b\*\* 0.40 ± 0.01a\*\*b\*\*

EU 0.59 ± 0.02 2.40 ± 0.07 1.473 ± 0.10 1.31 ± 0.01 0.60 ± 0.01 EU + B 0.55 ± 0.01 2.39 ± 0.09 1.32 ± 0.06 1.32 ± 0.01 0.60 ± 0.01 H 0.26 ± 0.01a\*\*\* 0.93 ± 0.03a\*\*\* 0.90 ± 0.03a\* 0.85 ± 0.01a\*\* 0.27 ± 0.05a\*\*\* H + B 0.35 ± 0.01a\*\*b\*\* 1.17 ± 0.01a\*\*b\*\* 1.07 ± 0.02a\*b\* 1.0 ± 0.01a\*b\* 0.40 ± 0.01a\*\*b\*\*

EU 0.57 ± 0.01 0.38 ± 0.01 0.78 ± 0.01 0.72 ± 0.01 0.47 ± 0.01 EU + B 0.56 ± 0.02 0.38 ± 0.01 0.76 ± 0.02 0.65 ± 0.05 0.50 ± 0.01 H 0.23 ± 0.01a\*\*\* 0.11 ± 0.01a\*\*\* 0.45 ± 0.01a\*\* 0.30 ± 0.01a \*\*\* 0.18 ± 0.01a\*\*\* H + B 0.35 ± 0.01a\*\*b\*\* 0.18 ± 0.01a\*\*b\* 0.54 ± 0.01a\*b\* 0.41 ± 0.01a\*\*b\*\* 0.28 ± 0.01a\*\*b\*\*

*a: Mean significance difference from control group. b: Mean significance difference from hypothyroid group.*

*Effect of barley on neurotransmitters level in discrete brain regions in control and treated groups.*

*DOI: http://dx.doi.org/10.5772/intechopen.92359*

excitatory amino acid (**Figures 9** and **10**).

**NE (μg g−1 tissue)**

**DA (μg g−1 tissue)**

**5-HT (μg g−1 tissue)**

*\*p < 0.05, \*\*p < 0.01 and \*\*\*p < 0.001.*

*All data in tables represented by mean ± SD, n = 10 animals.*

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility… DOI: http://dx.doi.org/10.5772/intechopen.92359*

Neurotransmitters NE, DA, and 5-HT levels were significantly reduced in all brain areas (cerebellum, midbrain, cerebral cortex, hypothalamus, and hippocampus) with hypothyroidism induction as shown in **Table 3**, these could be attributed to the reduced oestradiol level as mentioned above [37, 38]. The administration of *Hordeum vulgare* (barley) improved the disturbances in the dopaminergic, serotonergic and noradrenergic pathways via two different mechanisms; first, the high phytosterol content modulates estrogen receptors (Erα and β) expression and elevate oestradiol levels. Second, barley is enriched with tryptophan and phenylalanine and could regulate the synthesis of 5-HT, DA and NE through the conversion of tryptophan to 5-hydroxytryptophan (5-HTP) to 5-HT and hydrolysis of phenylalanine to generate tyrosine that ultimately produces DA and NE [2, 36, 39, 40]. In the present study, hypothyroidism induced a significant increase in inhibitory amino acid, including GABA and histidine, which is an excitatory amino acid (**Figures 9** and **10**).

These results could explain the increase in Caspase-3, which may be attributed to reduced blood oxygen–glucose levels in several brain regions as a result of increased GABA levels. The study also revealed an increase in dopamine receptors, whereas serotonin receptors were significantly decreased. The *Hordeum vulgare* (barley) treatment in the present study caused a renormalization the observed disturbances in the amino acid and neurotransmitter levels because it is enriched with folic acid, which is involved in the synthesis of monoamine neurotransmitters and modulate serotonergic, dopaminergic and noradrenergic systems by acting as a cofactor for enzymes that convert tryptophan to 5-HT and enzymes that convert tyrosine to noradrenaline [19]. The alteration in serotonin receptor densities was restored by the barley administration, due to its enriched levels of tryptophan, which is metabolized to serotonin [36, 40] and activates these receptors (**Figure 11**).


*All data in tables represented by mean ± SD, n = 10 animals.*

*\*p < 0.05, \*\*p < 0.01 and \*\*\*p < 0.001.*

*a: Mean significance difference from control group. b: Mean significance difference from hypothyroid group.*

### **Table 3.**

*Effect of barley on neurotransmitters level in discrete brain regions in control and treated groups.*

*Plant Stress Physiology*

**Figure 6.**

**Figure 7.**

*Effect of barley on caspase-3 (ng/ml) and 8-OhdG (ng/ml)in brain tissue.*

*DNA (B) tail length expressed in μm in thyroid tissue of all treated groups.*

*Effect of barley on DNA damage in thyroid tissue (A) fluorescence photomicrograph showing comets in EU-, H-, EU + B and H + B-groups. The indicated the intact DNA and indicated the degree of damaged* 

*Effect of barley on DNA damage in whole brain tissue (A) fluorescence photomicrograph showing comets in EU-, H-, EU + B and H + B-groups. The indicated the intact DNA and indicated the degree of damaged* 

*DNA (B) tail length expressed in μm in whole brain tissue of all treated groups.*

**356**

**Figure 8.**

**Figure 9.**

*Effect of barley on inhibitory amino acids in EU and H groups. All data represented by mean ± SD, n= 10 animals, \*p<0.05, \*\*p<0.01 and \*\*\*p<0.001, (a) mean significance difference from control group, (b) mean significance difference from hypothyroid group. (C) mean significance difference from EU+B.*

### **Figure 10.**

*Effect of barley on excitatory amino acids in EU and H groups. All data represented by mean ± SD, n= 10 animals, \*p<0.05, \*\*p<0.01 and \*\*\*p<0.001, (a) mean significance difference from control group. (B) mean significance difference from hypothyroid group. (C) mean significance difference from EU+B.*

The elevation in serotonin levels after barley administration in the present study also resulted in ERK1/2 improvement in brain tissue, which was reduced by hypothyroidism induction. The binding of serotonin to 5-TH2 receptors stimulates ERK1/2 phosphorylation via the release of epidermal growth factor (EGF) agonist

**359**

transmission [42].

**Figure 11.**

mood and depression.

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility…*

and transactivation of (EFG) receptors [41]. The improvement of serum oestradiol in hypothyroid-barley-treated groups as mentioned before could explain the positive effect of barley on restoring dopamine levels in brain tissues because the reproductive hormones, estrogen and progesterone, modulate the dysregulated serotonergic, dopaminergic, and glutamatergic neurotransmission by regulating the expression of receptors, the synthesis, reuptake, and release of the neu-

*Effect of barley on relative expression of (A) mRNA 5-HT1A, (B) mRNA 5-HT2A and (C) mRNA DA2A.*

rotransmitter serotonin and dopamine, which interact with dopaminergic neurons directly to downregulate D2 autoreceptors and indirectly by inhibiting GABAergic

Based on the above findings, we conclude that barley (*Hordeum vulgare*) is a nutritious food with high carbohydrate, zinc, magnesium content, and a high amino acids Trp:BCAA ratio has a positive effect on ameliorating the neural dysfunction induced by hypothyroidism and recommended for relieving stress, improving

*DOI: http://dx.doi.org/10.5772/intechopen.92359*

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility… DOI: http://dx.doi.org/10.5772/intechopen.92359*

and transactivation of (EFG) receptors [41]. The improvement of serum oestradiol in hypothyroid-barley-treated groups as mentioned before could explain the positive effect of barley on restoring dopamine levels in brain tissues because the reproductive hormones, estrogen and progesterone, modulate the dysregulated serotonergic, dopaminergic, and glutamatergic neurotransmission by regulating the expression of receptors, the synthesis, reuptake, and release of the neurotransmitter serotonin and dopamine, which interact with dopaminergic neurons directly to downregulate D2 autoreceptors and indirectly by inhibiting GABAergic transmission [42].

Based on the above findings, we conclude that barley (*Hordeum vulgare*) is a nutritious food with high carbohydrate, zinc, magnesium content, and a high amino acids Trp:BCAA ratio has a positive effect on ameliorating the neural dysfunction induced by hypothyroidism and recommended for relieving stress, improving mood and depression.

*Plant Stress Physiology*

**358**

**Figure 10.**

**Figure 9.**

The elevation in serotonin levels after barley administration in the present study also resulted in ERK1/2 improvement in brain tissue, which was reduced by hypothyroidism induction. The binding of serotonin to 5-TH2 receptors stimulates ERK1/2 phosphorylation via the release of epidermal growth factor (EGF) agonist

*difference from hypothyroid group. (C) mean significance difference from EU+B.*

*Effect of barley on excitatory amino acids in EU and H groups. All data represented by mean ± SD, n= 10 animals, \*p<0.05, \*\*p<0.01 and \*\*\*p<0.001, (a) mean significance difference from control group. (B) mean significance* 

*Effect of barley on inhibitory amino acids in EU and H groups. All data represented by mean ± SD, n= 10 animals, \*p<0.05, \*\*p<0.01 and \*\*\*p<0.001, (a) mean significance difference from control group, (b) mean* 

*significance difference from hypothyroid group. (C) mean significance difference from EU+B.*

### **4. The antioxidant effect of** *Panax ginseng* **on fertility disorders and oxidative stress induced by hypothyroidism**

The present study revealed a reduction in fT3, fT4, and elevation in TSH levels as well as deterioration in THs transporting protein (TBG and TTR) in brain tissues of hypothyroid rats as shown in **Table 8**. This disturbance attributed to Neo-Mercazole which is an antithyroid agent that blocks thyroid hormonogenesis by inhibiting thyroid peroxidase (TPO) activity and preventing the formation of thyroglobulin from tyrosine [43, 44]. Hypothyroidism also causes elevation of cortisol that leads to inhibition of the deiodinase enzyme type 2 (D2) enzyme, responsible for the conversion of T4 into T3 [45]. Ginseng treatment improves the levels of thyroid hormones in serum and brain tissues through restoration of the impairment transporting protein (TTR & TBG) as shown in **Tables 4** and **8**. Moreover, ginseng boosts the activity of the enzyme responsible for converting T4 to active T3 and reduces thyroid hormone-blocking reverse T3 (rT3) which inhibits active T3 from binding to its functioning T3 receptors [46, 47].

The sexual dysfunction may be developed from psychological stress state exerted by hypothyroidism induction and this confirmed the role of the pituitary– adrenal gonadal axis (HPA) as a defense mechanism carried out by the organism against stress event [48]. So the reduction in trophic hormone (FSH & LH) associated with hypothyroidism led to decreasing the E2 hormone level and elevation of progesterone and testosterone. The inhibition in gonadal activity in hypothyroid rats in the present model as documented in **Table 5** was confirmed by the lowering of E2/T (**Table 6**) ratio which is a marker of the aromatase enzyme activity


*All data in tables represented by mean ± SD, n= 10 animals.*

*\*p<0.05, \*\*p<0.01 and \*\*\*p<0.001.*

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

### **Table 4.**

*Thyroid hormones and their carrying proteins levels of in the studied groups.*


*All data in tables represented by mean ± SD, n= 10 animals.*

*\*p<0.05, \*\*p<0.01 and \*\*\*p<0.001.*

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

**361**

**Table 8.**

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility…*

(estrogen synthase, CYP19A1), this is a key enzyme converted testosterone into estrogen in granulosa cell [49]. Hyperprolactinemia, as recorded in the present study, negatively affected the activity of aromatase enzyme and this lead to hypoestrogenism, inhibited the release of gonadotrophic hormones (LH & FSH) from the pituitary gland and potentiated the inhibitory action of inhibin hormone that stimulated negative feedback and lowered estradiol level [50]. The administration of *Panax ginseng* was ameliorated these changes in trophic and steroidal gonadal hormones due to the triterpenoid saponins which steroidal in nature and considered

The current study exhibited a significant increase in serum corticosterone hormone. Also, the elevation of 8-hydroxyguanosine, an oxidative stress marker, and Caspase-3, an apoptotic marker in serum and brain tissues (**Table 7** and **Figure 12**). The administration of *Panax ginseng* was renormalized the cortisol and oxidative stress markers by increasing the cellular resistance to stress and potentiated the role

**Parameter Control G H H+ G** ERK1/2 (pg/ml) 47.38 ± 1.93 50.17 ± 2.57 63.17 ± 3.13 (a\*) 42.26 ± 1.7 (a\*b\*\*) E2/T ratio 64.87 ± 10.3 58.0 ± 13.0 37.6 ± 11.4(a\*\*\*) 51.4 ± 19.25 (b\*) Corticosterone (μg/dl) 1.8 ± 0.06 1.7 ± 0.11 2.82 ± 0.21 (a\*\*) 1.73 ± 0.10 (b\*)

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

**Parameter Control G H H+ G** 8-OH Guanosine (ng/L) 48.46 ± 2.21 32.79 ± 0.05 57.99 ± 0.02 (a\*) 48.63 ± 0.08 (b\*) Caspases (ng/ml) 5.26 ± 0.20 5.94 ± 0.15 12.44 ± 0.67 (a\*\*\*) 5.48 ± 0.14 (b\*\*\*)

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

(a\*\*\*)

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

*Effect of ginseng on fT3, fT4, TSH, TBG and TTR in brain tissue of control and hypothyroid-treated rats.*

f T4 (μg/ml) 4.44 ± 0.11 3.97 ± 0.10 (a\*) 3.65 ± 0.34 (a\*b\*) 4.08 ± 0.58 (c\*) f T3 (μg/ml) 1.62 ± 0.08 1.40 ± 0.08 (a\*) 1.49 ± 0.12 (a\*b\*) 1.51 ± 0.11 (a\*b\*)

TBG (pg/ml) 1.58 ± 0.04 2.09 ± 0.08 (a\*) 1.75 ± 0.39 (a\*b\*) 1.73 ± 0.35 (a\*b\*)

**Control H G H+ G**

(a\*\*\*b\*\*\*)

42.1 ± 2.88 (a\*\*b\*\*\*)

13.57 ± 0.87 (b\*\*\*c\*\*\*)

33.28 ± 1.88 (a\*\*\*b\*\*\*c\*\*)

*Oxidative stress markers in serum of hypothyroid and ginseng-treated female albino rats.*

TSH (mIU/ml) 12.47 ± 0.42 15.81 ± 0.53 (a\*\*) 10.87 ± 0.48

*ERK1/2, E2/T ratio and cortisol level in serum of hypothyroid and treated female albino rats.*

the precursors of steroidal hormones in both plants and animals [51].

*DOI: http://dx.doi.org/10.5772/intechopen.92359*

*All data in tables represented by mean ± SD, n= 10 animals.*

*All data in tables represented by mean ± SD, n= 10 animals.*

TTR (ng/ml) 51.23 ± 2.39 15.07 ± 0.14

*All data in tables represented by mean ± SD, n= 10 animals.*

*\*p<0.05, \*\*p<0.01 and \*\*\*p<0.001.*

*\*p<0.05, \*\*p<0.01 and \*\*\*p<0.001.*

*\*p<0.05, \*\*p<0.01 and \*\*\*p<0.001.*

**Table 6.**

**Table 7.**

### **Table 5.**

*Serum fertility hormones levels in hypothyroid and treated adult female albino rats.*

### *Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility… DOI: http://dx.doi.org/10.5772/intechopen.92359*

(estrogen synthase, CYP19A1), this is a key enzyme converted testosterone into estrogen in granulosa cell [49]. Hyperprolactinemia, as recorded in the present study, negatively affected the activity of aromatase enzyme and this lead to hypoestrogenism, inhibited the release of gonadotrophic hormones (LH & FSH) from the pituitary gland and potentiated the inhibitory action of inhibin hormone that stimulated negative feedback and lowered estradiol level [50]. The administration of *Panax ginseng* was ameliorated these changes in trophic and steroidal gonadal hormones due to the triterpenoid saponins which steroidal in nature and considered the precursors of steroidal hormones in both plants and animals [51].

The current study exhibited a significant increase in serum corticosterone hormone. Also, the elevation of 8-hydroxyguanosine, an oxidative stress marker, and Caspase-3, an apoptotic marker in serum and brain tissues (**Table 7** and **Figure 12**). The administration of *Panax ginseng* was renormalized the cortisol and oxidative stress markers by increasing the cellular resistance to stress and potentiated the role


*All data in tables represented by mean ± SD, n= 10 animals.*

*\*p<0.05, \*\*p<0.01 and \*\*\*p<0.001.*

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

### **Table 6.**

*Plant Stress Physiology*

**4. The antioxidant effect of** *Panax ginseng* **on fertility disorders** 

The present study revealed a reduction in fT3, fT4, and elevation in TSH levels as well as deterioration in THs transporting protein (TBG and TTR) in brain tissues of hypothyroid rats as shown in **Table 8**. This disturbance attributed to Neo-Mercazole which is an antithyroid agent that blocks thyroid hormonogenesis by inhibiting thyroid peroxidase (TPO) activity and preventing the formation of thyroglobulin from tyrosine [43, 44]. Hypothyroidism also causes elevation of cortisol that leads to inhibition of the deiodinase enzyme type 2 (D2) enzyme, responsible for the conversion of T4 into T3 [45]. Ginseng treatment improves the levels of thyroid hormones in serum and brain tissues through restoration of the impairment transporting protein (TTR & TBG) as shown in **Tables 4** and **8**. Moreover, ginseng boosts the activity of the enzyme responsible for converting T4 to active T3 and reduces thyroid hormone-blocking reverse T3 (rT3) which inhibits active T3 from binding to its functioning T3 receptors [46, 47]. The sexual dysfunction may be developed from psychological stress state exerted by hypothyroidism induction and this confirmed the role of the pituitary– adrenal gonadal axis (HPA) as a defense mechanism carried out by the organism against stress event [48]. So the reduction in trophic hormone (FSH & LH) associated with hypothyroidism led to decreasing the E2 hormone level and elevation of progesterone and testosterone. The inhibition in gonadal activity in hypothyroid rats in the present model as documented in **Table 5** was confirmed by the lowering of E2/T (**Table 6**) ratio which is a marker of the aromatase enzyme activity

**Parameter Control G H H+ G** f T3 (ng/ml) 1.99 ± 0.06 1.54 ± 0.05 1.59 ± 0.02 1.80 ± 0.04 (b\*) f T4 (μg/ml) 6.61 ± 0.14 7.32 ± 0.36 4.07 ± 0.12 (a\*) 6.78 ± 0.34 (b\*\*) TSH (mIU/ml) 12.31 ± 0.62 12.81 ± 0.69 14.09 ± 0.42 (a\*) 10.75 ± 0.26 (b\*\*) TTR (ng/ml) 32.56 ± 2.04 36.93 ± 2.95 44.19 ± 3.34 (a\*) 27.41 ± 1.96 (b\*) TBG(pg/ml) 2.54 ± 0.19 2.49 ± 0.06 2.26 ± 0.07 2.37 ± 0.13

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

**Parameter Control G H H+ G** FSH (mIU/L) 7.11 ± 0.16 7.98 ± 0.14 5.99 ± 0.18 (a\*) 7.03 ± 0.12 LH (ng/L) 6.47 ± 0.29 7.38 ± 0.38 4.35 ± 0.19 (a\*\*) 6.73 ± 0.28 (b\*\*) E2 (Pg/ml) 20.76 ± 0.64 20.3 ± 0.39 14.7 ± 0.8 (a\*) 18.00 ± 0.77 (a\*,b\*\*) P (ng/ml) 23.24 ± 1.66 20.66 ± 1.51 30.41 ± 1.18 (a\*) 20.56 ± 1.67 (b\*) T (ng/ml) 0.32 ± 0.06 0.35 ± 0.03 0.39 ± 0.07 (a\*) 0.35 ± 0.04 PRL (ng/ml) 94.06 ± 4.89 104.12 ± 4.41 149.11 ± 11.33 (a\*\*) 101.02 ± 6.58 (b\*)

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

*Serum fertility hormones levels in hypothyroid and treated adult female albino rats.*

*All data in tables represented by mean ± SD, n= 10 animals.*

*All data in tables represented by mean ± SD, n= 10 animals.*

*\*p<0.05, \*\*p<0.01 and \*\*\*p<0.001.*

*Thyroid hormones and their carrying proteins levels of in the studied groups.*

*\*p<0.05, \*\*p<0.01 and \*\*\*p<0.001.*

**Table 4.**

**and oxidative stress induced by hypothyroidism**

**360**

**Table 5.**

*ERK1/2, E2/T ratio and cortisol level in serum of hypothyroid and treated female albino rats.*


*All data in tables represented by mean ± SD, n= 10 animals.*

*\*p<0.05, \*\*p<0.01 and \*\*\*p<0.001.*

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

### **Table 7.**

*Oxidative stress markers in serum of hypothyroid and ginseng-treated female albino rats.*


*All data in tables represented by mean ± SD, n= 10 animals.*

*\*p<0.05, \*\*p<0.01 and \*\*\*p<0.001.*

*a: mean significance difference from control group. B: mean significance difference from hypothyroid group.*

### **Table 8.**

*Effect of ginseng on fT3, fT4, TSH, TBG and TTR in brain tissue of control and hypothyroid-treated rats.*

**Figure 12.** *Effect of ginseng on ERK1/2, Caspase-3 and 8-OHdG in all studied groups.*

**363**

**Figure 14.**

*Effect of ginseng on inhibitory amino acids in all studied groups.*

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility…*

of the immune system through triterpenes of ginsenosides [48, 52]. Serum ERK1/2 was activated in response to the elevation of oxidative stress and cell death apoptotic markers as underlined in the present study. Treatment with ginseng ameliorates 8-OHdG, Caspase-3 and ERK1/2 levels referring to its neuroprotective effect and retrieval homeostasis. Also, ginsenosides which is the pharmacologically active constituents with its adaptogenic, powerful antioxidant and radical scavenging activities, regulate the function of HPA, support neurogenesis, synaptogenesis, neuronal growth, and neurotransmission, in turn, protect the central nervous system [53–56]. The study of [57] showed that the panaxatriol group of ginsenosides blocked Caspase-3 expression and increased anti-apoptotic Bcl-2 and p53, indicating that RG repressed cellular apoptosis otherwise, ginsenosides Rd and Re have neuroprotective properties by modulation of ERK1/2 signaling pathway [58]. The elevation of Caspase-3 in the present hypothyroid modal was confirmed by studying the Comet tailed DNA damage of the brain and thyroid tissues as illustrated in **Figures 15** and **16**. Ginseng treatment repair DNA damage in brain and thyroid tissues, this denoted to

its highest content of phenolic compounds which act as antioxidants.

genesis, maturation and synaptic transmission (**Figures 13** and **14**).

Thyroid hormones control the levels of these neurotransmitters which are responsible for maintaining a good mental state and preventing depression [19]. THs regulate both the release of the neurotransmitters and their post-receptor signaling to promote mood stabilization so, their deficiency may weaken the neuro-

The present data, exhibited that the induction of hypothyroidism resulted in a significant decrease of NE, DA and 5-HT concentrations in all studied brain areas

*DOI: http://dx.doi.org/10.5772/intechopen.92359*

**Figure 13.** *Effect of ginseng on excitatory amino acids in all studied groups.*

### *Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility… DOI: http://dx.doi.org/10.5772/intechopen.92359*

of the immune system through triterpenes of ginsenosides [48, 52]. Serum ERK1/2 was activated in response to the elevation of oxidative stress and cell death apoptotic markers as underlined in the present study. Treatment with ginseng ameliorates 8-OHdG, Caspase-3 and ERK1/2 levels referring to its neuroprotective effect and retrieval homeostasis. Also, ginsenosides which is the pharmacologically active constituents with its adaptogenic, powerful antioxidant and radical scavenging activities, regulate the function of HPA, support neurogenesis, synaptogenesis, neuronal growth, and neurotransmission, in turn, protect the central nervous system [53–56]. The study of [57] showed that the panaxatriol group of ginsenosides blocked Caspase-3 expression and increased anti-apoptotic Bcl-2 and p53, indicating that RG repressed cellular apoptosis otherwise, ginsenosides Rd and Re have neuroprotective properties by modulation of ERK1/2 signaling pathway [58]. The elevation of Caspase-3 in the present hypothyroid modal was confirmed by studying the Comet tailed DNA damage of the brain and thyroid tissues as illustrated in **Figures 15** and **16**. Ginseng treatment repair DNA damage in brain and thyroid tissues, this denoted to its highest content of phenolic compounds which act as antioxidants.

Thyroid hormones control the levels of these neurotransmitters which are responsible for maintaining a good mental state and preventing depression [19]. THs regulate both the release of the neurotransmitters and their post-receptor signaling to promote mood stabilization so, their deficiency may weaken the neurogenesis, maturation and synaptic transmission (**Figures 13** and **14**).

The present data, exhibited that the induction of hypothyroidism resulted in a significant decrease of NE, DA and 5-HT concentrations in all studied brain areas

**Figure 14.** *Effect of ginseng on inhibitory amino acids in all studied groups.*

*Plant Stress Physiology*

**362**

**Figure 13.**

**Figure 12.**

*Effect of ginseng on excitatory amino acids in all studied groups.*

*Effect of ginseng on ERK1/2, Caspase-3 and 8-OHdG in all studied groups.*


**Table 9.**

**365**

**Figure 16.**

**Figure 15.**

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility…*

(frontal cortex, hippocampus, hypothalamus, midbrain and cerebellum) as shown in **Table 9**. Monoamines reduction after hypothyroidism refereed to the disturbance in the synthesis and release of these amines from impairment neurons or may be due to an alteration pattern of their synthesizing and/or degradative enzymes [59]. Ginseng treatment ameliorate the reduced monoamine levels of hypothyroid rats and this refers to its powerful ability to maintain homeostasis and modulating neurotransmitter levels hence can amend the neurodegenerative diseases [54, 60]. Also, ginseng saponins modulate dopaminergic activity at both presynaptic and

Improvement of monoamines after ginseng treatment refer also to gintonin which is one of the important ginseng constituents that increased the expres

sion of learning and memory and modulate cholinergic, glutaminergic and other molecular signaling pathways that are vital for cognitive activity as stated in [16]. Ginseng which considered a potential phytoestrogen exhibits antidepressant so, ginsenosides Rb1 enhances the serotonergic system by increasing 5-HT synthesis,

*Effect of ginseng on DNA damage in whole brain tissue, (A) Tail length expressed in μm in brain tissue of all treated groups. \*Significant at p. 0.05, \*\*significant at p. 0.01 and \*\*\*significant at p. 0.001. (a), significant versus control group, (b) significant versus hypothyroid, (H) group and (c ) significant versus ginseng, (G) treated group, (B) Fluorescence photomicrograph showing comets in control, H, G and H+G -groups. (blue arrows) indicated the intact DNA and (green arrows) indicated the degree of damaged DNA (tail).* 

*Effect of ginseng on DNA damage in thyroid tissue. (A) Tail length expressed in μm in thyroid tissue of all treated groups. \*\* significant at p 0.01 and \*\*\* significant at p 0.001. (a) significant versus control group, (b) significant versus hypothyroid, (H), group and (c ), significant versus ginseng (G), treated group (B), Fluorescence photomicrograph showing comets in control, H, G and H+G -groups. (blue arrows) indicated the* 

*intact DNA and (green arrows) indicated the degree of damaged DNA (tail).*


*DOI: http://dx.doi.org/10.5772/intechopen.92359*

postsynaptic receptors [54, 55].

*Effect of ginseng on monoamines levels of discrete brain regions in control and hypothyroid-treated rats.*

### *Plant Stress Physiology*

### *Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility… DOI: http://dx.doi.org/10.5772/intechopen.92359*

(frontal cortex, hippocampus, hypothalamus, midbrain and cerebellum) as shown in **Table 9**. Monoamines reduction after hypothyroidism refereed to the disturbance in the synthesis and release of these amines from impairment neurons or may be due to an alteration pattern of their synthesizing and/or degradative enzymes [59]. Ginseng treatment ameliorate the reduced monoamine levels of hypothyroid rats and this refers to its powerful ability to maintain homeostasis and modulating neurotransmitter levels hence can amend the neurodegenerative diseases [54, 60]. Also, ginseng saponins modulate dopaminergic activity at both presynaptic and postsynaptic receptors [54, 55].

Improvement of monoamines after ginseng treatment refer also to gintonin which is one of the important ginseng constituents that increased the expression of learning and memory and modulate cholinergic, glutaminergic and other molecular signaling pathways that are vital for cognitive activity as stated in [16]. Ginseng which considered a potential phytoestrogen exhibits antidepressant so, ginsenosides Rb1 enhances the serotonergic system by increasing 5-HT synthesis,

### **Figure 15.**

*Plant Stress Physiology*

**364**

**Frontal cortex**

> **DA (μg g−1 tissue)**

Control

H G H + G **NE (μg g−1 tissue)**

Control

H G H + G **5-HT (μg g−1 tissue)**

Control

H G H + G

0.57 ± 0.03 0.23 ± 0.01 a\*\*\*

0.57 ± 0.03b\*\*\*

0.41 ± 0.02a\*\*\*b\*\*\*c\*\*\*

*Data in tables given are mean ± S.D. the number of animals was 10 in each group.*

*\*\*Significant at p 0.01 and \*\*\*significant at p 0.001.*

**Table 9.**

*(a) Significant versus control group; (b) significant versus hypothyroid (H) group and (c) significant versus ginseng (G) treated group.*

*Effect of ginseng on monoamines levels of discrete brain regions in control and hypothyroid-treated rats.*

0.38 ± 0.02 0.11 ± 0.01 a\*\*\*

0.39 ± 0.01b\*\*\*

0.29 ± 0.01a\*\*\*b\*\*\*c\*\*\*

0.78 ± 0.03 0.45 ± 0.02 a\*\*\*

0.77 ± 0.02b\*\*\*

0.56 ± 0.02a\*\*\*b\*\*\*c\*\*\*

0.72 ± 0.03 0.29 ± 0.01 a\*\*\*

0.70 ± 0.03b\*\*\*

0.5 ± 0.01a\*\*\*b\*\*\*c\*\*\*

0.47 ± 0.02

0.18 ± 0.01 a\*\*\*

0.47 ± 0.01b\*\*\*

0.34 ± 0.01a\*\*\*b\*\*\*c\*\*\*

0.51 ± 0.07 0.21 ± 0.02 a\*\*\*

0.56 ± 0.08b\*\*\*

0.35 ± 0.03a\*\*\*b\*\*c\*\*\*

0.68 ± 0.04 0.31 ± 0.02 a\*\*\*

0.67 ± 0.06 b\*\*\*

0.54 ± 0.03a\*\*\*b\*\*\*c\*\*\*

0.39 ± 0.01 0.13 ± 0.02 a\*\*\*

0.38 ± 0.02b\*\*\*

0 .29 ± 0.01a\*\*\*b\*\*\*c\*\*\*

0.70 ± 0.01 0.35 ± 0.02 a\*\*\*

0.69 ± 0.01b\*\*\*

0.56 ± 0.03a\*\*\*b\*\*\*c\*\*\*

0.58 ± 0.03

0.29 ± 0.01 a\*\*\*

0.6 ± 0.01b\*\*\*

0.49 ± 0.01a\*\*\*b\*\*\*c\*\*\*

0.59 ± 0.04 0.25 ± 0.02 a\*\*\*

0.58 ± 0.01b\*\*\*

0.44 ± 0.02a\*\*\*b\*\*c\*\*\*

2.39 ± 0.18 0.93 ± 0.07 a\*\*\*

2.74 ± 0.08b\*\*\*

1.67 ± 0.07a\*\*\*b\*\*\*c\*\*\*

1.47 ± 0.27 0.89 ± 0.08 a\*\*\*

1.69 ± 0.17b\*\*\* 1.12 ± 0.04a\*\*c\*\*\*

1.31 ± 0.01 0.89 ± 0.03 a\*\*\*

1.31 ± 0.01b\*\* 1.18 ± 0.01a\*\*\*b\*\*\*c\*\*\*

0.60 ± 0.02

0.27 ± 0.01 a\*\*\*

0.59 ± 0.01b\*\*\*

0.50 ± 0.01a\*\*b\*\*\*c\*\*

**Hippocampus**

**Hypothalamus**

**Mid brain**

**Cerebellum**

*Effect of ginseng on DNA damage in whole brain tissue, (A) Tail length expressed in μm in brain tissue of all treated groups. \*Significant at p. 0.05, \*\*significant at p. 0.01 and \*\*\*significant at p. 0.001. (a), significant versus control group, (b) significant versus hypothyroid, (H) group and (c ) significant versus ginseng, (G) treated group, (B) Fluorescence photomicrograph showing comets in control, H, G and H+G -groups. (blue arrows) indicated the intact DNA and (green arrows) indicated the degree of damaged DNA (tail).* 

### **Figure 16.**

*Effect of ginseng on DNA damage in thyroid tissue. (A) Tail length expressed in μm in thyroid tissue of all treated groups. \*\* significant at p 0.01 and \*\*\* significant at p 0.001. (a) significant versus control group, (b) significant versus hypothyroid, (H), group and (c ), significant versus ginseng (G), treated group (B), Fluorescence photomicrograph showing comets in control, H, G and H+G -groups. (blue arrows) indicated the intact DNA and (green arrows) indicated the degree of damaged DNA (tail).*

### **Figure 17.**

*Effect of ginseng on expression of mRNA 5-HT1A, mRNA 5-HT2A and mRNA DA2A. \*Significant at p 0.05, and \*\*\*significant at p 0.001. (a) Significant versus control group (b) significant versus hypothyroid (H) group and (c) significant versus ginseng (G) treated group.*

decreasing 5-HT degradation, stimulating 5-HT2A receptor and suppress the activity of the inhibitory 5-HT3A receptor in the brain. Also, effect via increasing 5-HT activity. This effect is mediated by the activation of estrogen receptor [61, 62]. In the present study, hypothyroidism induction by Neo-Mericazole lead to a significant increase of excitatory amino acid, histidine, and all inhibitory amino acids while excitatory glutamic acid was significantly decreased in brain tissues (**Figures 15** and **16**). *Panax ginseng* with its powerful components ameliorates the disturbance of amino acids and in turn monoamines. The induction of hypothyroidism revealed an elevation in the concentration of dopamine receptors and the reduction of serotonin receptors density (**Figure 17**). Treatment with ginseng restores the level of dopamine and serotonin receptors density towards the control value. This refers firstly to the genomic pathway of ginsenosides which bind to intracellular nuclear hormone receptors like androgen receptor (AR), estrogenic receptor (ER) and progesterone receptor [13].

### **5. Conclusion**

In conclusion, the present study is pointed out to the pituitary-gonad-adrenal disturbances aroused from the hypothyroidism induction by Neo-Mericazole and how ginseng, one of the most Asian medicinal traditional plants, significantly normalized the fertility disorders and stress by acting as free radicals' scavenger.

**367**

**Author details**

Lobna F. Wahman1

and Magda H.M. Yousef2,3

\*, Marwa M. Abd Rabo1

2 Faculty of Medicine, Taibah University, Saudi Arabia

\*Address all correspondence to: lwahman62@gmail.com

3 Faculty of Medicine, Ain-Shams University, Egypt

provided the original work is properly cited.

1 National Organization for Drug Control and Research (NODCAR), Egypt

© 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,

, Amany Hanafy M. Elgoly1

*Potential Role of Plants Hordeum vulgare L. and Panax ginseng L. in Resolving the Fertility…*

*DOI: http://dx.doi.org/10.5772/intechopen.92359*

The authors declare no conflict of interest.

**Conflict of interest**

*Potential Role of Plants Hordeum vulgare L. and Panax ginseng L. in Resolving the Fertility… DOI: http://dx.doi.org/10.5772/intechopen.92359*

## **Conflict of interest**

*Plant Stress Physiology*

decreasing 5-HT degradation, stimulating 5-HT2A receptor and suppress the activity of the inhibitory 5-HT3A receptor in the brain. Also, effect via increasing 5-HT activity. This effect is mediated by the activation of estrogen receptor [61, 62]. In the present study, hypothyroidism induction by Neo-Mericazole lead to a significant increase of excitatory amino acid, histidine, and all inhibitory amino acids while excitatory glutamic acid was significantly decreased in brain tissues (**Figures 15** and **16**). *Panax ginseng* with its powerful components ameliorates the disturbance of amino acids and in turn monoamines. The induction of hypothyroidism revealed an elevation in the concentration of dopamine receptors and the reduction of serotonin receptors density (**Figure 17**). Treatment with ginseng restores the level of dopamine and serotonin receptors density towards the control value. This refers firstly to the genomic pathway of ginsenosides which bind to intracellular nuclear hormone receptors like androgen receptor (AR), estrogenic

*Effect of ginseng on expression of mRNA 5-HT1A, mRNA 5-HT2A and mRNA DA2A. \*Significant at p 0.05, and \*\*\*significant at p 0.001. (a) Significant versus control group (b) significant versus hypothyroid (H) group* 

In conclusion, the present study is pointed out to the pituitary-gonad-adrenal disturbances aroused from the hypothyroidism induction by Neo-Mericazole and how ginseng, one of the most Asian medicinal traditional plants, significantly normalized the fertility disorders and stress by acting as free radicals' scavenger.

receptor (ER) and progesterone receptor [13].

*and (c) significant versus ginseng (G) treated group.*

**366**

**5. Conclusion**

**Figure 17.**

The authors declare no conflict of interest.

### **Author details**

Lobna F. Wahman1 \*, Marwa M. Abd Rabo1 , Amany Hanafy M. Elgoly1 and Magda H.M. Yousef2,3

1 National Organization for Drug Control and Research (NODCAR), Egypt


\*Address all correspondence to: lwahman62@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.

### **References**

[1] Gangopadhyay N, Hossain MB, Rai DK, Brunton NP. A review of extraction and analysis of bioactives in oat and barley and scope for use of novel food processing technologies. Molecules. 2015;**20**:10884-10909

[2] Youssef MKE, El-fishawy FAE, Ramadan EAE. Nutritional assessment of barley, talbina and their germinated products. Scientific Journal of Crop Science. 2013;**3**:8-19

[3] Rahman MH, Ali MY. The relationships between thyroid hormones and the brain serotonin (5-Ht) system and mood: Of synergy and significance in the adult brain—A review. Faridpur Medical College Journal. 2015;**9**:98-101

[4] Zhu Y, Li T, Fu X, Abbasi AM, Zheng B, Liu RH. Phenolics content, antioxidant and antiproliferative activities of dehulled highland barley (*Hordeum vulgare* L.). Journal of Functional Foods. 2015;**19**:439-450

[5] Idehen E, Tang Y, Sang S. Bioactive phytochemicals in barley. Journal of Food and Drug Analysis. 2017;**25**: 148-161

[6] Choi JS et al. Consumption of barley β-glucan ameliorates fatty liver and insulin resistance in mice fed a highfat diet. Molecular Nutrition & Food Research. 2010;**54**:1004-1013

[7] Jonnalagadda SS et al. Putting the whole grain puzzle together: Health benefits associated with whole grains—Summary of American Society for Nutrition 2010 Satellite Symposium. The Journal of Nutrition. 2011;**141**:1011S–1022S

[8] Schlemmer U, Frølich W, Prieto RM, Grases F. Phytate in foods and significance for humans: Food sources, intake, processing, bioavailability, protective role and analysis. Molecular

Nutrition & Food Research. 2009;**53**: S330–S375

[9] Yeo HB, Yoon HK, Lee HJ, Kang SG, Jung KY, Kim L. Effects of Korean Red Ginseng on cognitive and motor function: A double-blind, randomized, placebo-controlled trial. Journal of Ginseng Research. 2012;**36**(2):190-197

[10] So S, Lee JW, Kim YS, Hyun SH, Han CK. Red ginseng monograph. Journal of Ginseng Research. 2018;**42**(4):549-561

[11] Kim J, Cho SY, Kim SH, Kim S, Park CW, Cho D, et al. Ginseng berry and its biological effects as a natural phytochemical. Natural Products Chemistry & Research. 2016;**4**(2):2-5

[12] Rokot NT, Kairupan TS, Cheng KC, Runtuwene J, Kapantow NH, Amitani M, et al. A role of ginseng and its constituents in the treatment of central nervous system disorders. Evidence-based Complementary and Alternative Medicine. 2016. ID 2614742

[13] Park J, Song H, Kim SK, Lee MS, Rhee DK, Lee Y. Effects of ginseng on two main sex steroid hormone receptors: Estrogen and androgen receptors. Journal of Ginseng Research. 2017;**41**(2):215-221

[14] Kim KH, Lee D, Lee HL, Kim CE, Jung K, Kang KS. Beneficial effects of Panax ginseng for the treatment and prevention of neurodegenerative diseases: Past findings and future directions. Journal of Ginseng Research. 2018;**42**(3):239-247

[15] Chung IM, Lim JJ, Ahn MS, Jeong HN, An TJ, Kim SH. Comparative phenolic compound profiles and antioxidative activity of the fruit, leaves, and roots of Korean ginseng (*Panax ginseng* Meyer) according to cultivation years. Journal of Ginseng Research. 2016;**40**(1):68-75

**369**

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility…*

the effects of experimentally induced hypothyroidism and hyperthyroidism in some brain regions in albino rats. International Journal of Developmental

Neuroscience. 2010;**28**:371-389

[25] Karmarkar SW, Bottum KM, Tischkau SA. Considerations for the use of anaesthetics in neurotoxicity studies.

Comparative Medicine. 2010;**60**:

[26] Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research. 1988;

[27] Pagel P, Blome J, Wolf HU. Highperformance liquid chromatographic separation and measurement of various biogenic compounds possibly involved in the pathomechanism of Parkinson's disease. Journal of Chromatography. B, Biomedical Sciences and Applications.

[28] Heinrikson RL, Meredith SC. Amino acid analysis by reverse-phase highperformance liquid chromatography: Precolumn derivatization with phenylisothiocyanate. Analytical Biochemistry. 1984;**136**(1):65-74

[29] Borg S et al. Iron transport, deposition and bioavailability in the wheat and barley grain. Plant and Soil.

[30] Sun Q et al. Effects of forced swimming stress on thyroid function, pituitary thyroid-stimulating hormone and hypothalamus thyrotropin releasing hormone expression in adrenalectomy

Wistar rats. Experimental and Therapeutic Medicine. 2016

Parasitology. 2013;**135**:110-115

[31] Faccio L et al. Serum levels of LH, FSH, estradiol and progesterone in female rats experimentally infected by *Trypanosoma evansi*. Experimental

256-262

**175**(1):184-191

2000;**46**(2):297-304

2009;**325**:15-24

*DOI: http://dx.doi.org/10.5772/intechopen.92359*

[16] Jakaria M, Haque MDE, Kim J, Cho D-Y, Kim I-S, Choi DK. Active ginseng components in cognitive impairment: Therapeutic potential and prospects for delivery and clinical study. Oncotarget.

2018;**9**(71):33601-33620

2007;**3**:249-259

2012;**31**:282-289

2013;**8**:e55599

2010;**5**:134-142

Chapter 6

1954;**14**:1230-1244

[17] Goel R, Rajesh R. A signaling network of thyroid-stimulating hormone. Journal of Proteomics & Bioinformatics. 2011;**04**:238-241

[18] Bernal J. Thyroid hormone receptors in brain development. Nature Clinical Practice. Endocrinology & Metabolism.

[19] Tousson E, Ibrahim W, Arafa N, Akela MA. Monoamine concentrations

[20] Wu S, Tan G, Dong X, Zhu Z, Li W, Lou Z, et al. Profiling provides a system understanding of hypothyroidism in rats and its application. PLoS One.

[21] Bawazir AE. Investigations on the chronic effect of Talbina (barley water) on hormone (cortisol and testosterone), reproductive system and some neurotransmitter. American Eurasian Journal of Scientific Research.

[22] Paget GE, Barnes JM. Toxicity tests: Evaluation of drug activities. In: Laurence DR, Bandrakoach AL, editors. Pharmacometrics. Vol. 1. 1964. p. 135.

[23] Fraser TR et al. Antithyroid activity and toxicity of mercazole and neomercazole. The Journal of Clinical Endocrinology and Metabolism.

[24] El-Bakry AM, El-Gareib AW, Ahmed RG. Comparative study of

changes in the PTU-induced hypothyroid rat brain and the ameliorating role of folic acid. Human & Experimental Toxicology. *Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility… DOI: http://dx.doi.org/10.5772/intechopen.92359*

[16] Jakaria M, Haque MDE, Kim J, Cho D-Y, Kim I-S, Choi DK. Active ginseng components in cognitive impairment: Therapeutic potential and prospects for delivery and clinical study. Oncotarget. 2018;**9**(71):33601-33620

[17] Goel R, Rajesh R. A signaling network of thyroid-stimulating hormone. Journal of Proteomics & Bioinformatics. 2011;**04**:238-241

[18] Bernal J. Thyroid hormone receptors in brain development. Nature Clinical Practice. Endocrinology & Metabolism. 2007;**3**:249-259

[19] Tousson E, Ibrahim W, Arafa N, Akela MA. Monoamine concentrations changes in the PTU-induced hypothyroid rat brain and the ameliorating role of folic acid. Human & Experimental Toxicology. 2012;**31**:282-289

[20] Wu S, Tan G, Dong X, Zhu Z, Li W, Lou Z, et al. Profiling provides a system understanding of hypothyroidism in rats and its application. PLoS One. 2013;**8**:e55599

[21] Bawazir AE. Investigations on the chronic effect of Talbina (barley water) on hormone (cortisol and testosterone), reproductive system and some neurotransmitter. American Eurasian Journal of Scientific Research. 2010;**5**:134-142

[22] Paget GE, Barnes JM. Toxicity tests: Evaluation of drug activities. In: Laurence DR, Bandrakoach AL, editors. Pharmacometrics. Vol. 1. 1964. p. 135. Chapter 6

[23] Fraser TR et al. Antithyroid activity and toxicity of mercazole and neomercazole. The Journal of Clinical Endocrinology and Metabolism. 1954;**14**:1230-1244

[24] El-Bakry AM, El-Gareib AW, Ahmed RG. Comparative study of the effects of experimentally induced hypothyroidism and hyperthyroidism in some brain regions in albino rats. International Journal of Developmental Neuroscience. 2010;**28**:371-389

[25] Karmarkar SW, Bottum KM, Tischkau SA. Considerations for the use of anaesthetics in neurotoxicity studies. Comparative Medicine. 2010;**60**: 256-262

[26] Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research. 1988; **175**(1):184-191

[27] Pagel P, Blome J, Wolf HU. Highperformance liquid chromatographic separation and measurement of various biogenic compounds possibly involved in the pathomechanism of Parkinson's disease. Journal of Chromatography. B, Biomedical Sciences and Applications. 2000;**46**(2):297-304

[28] Heinrikson RL, Meredith SC. Amino acid analysis by reverse-phase highperformance liquid chromatography: Precolumn derivatization with phenylisothiocyanate. Analytical Biochemistry. 1984;**136**(1):65-74

[29] Borg S et al. Iron transport, deposition and bioavailability in the wheat and barley grain. Plant and Soil. 2009;**325**:15-24

[30] Sun Q et al. Effects of forced swimming stress on thyroid function, pituitary thyroid-stimulating hormone and hypothalamus thyrotropin releasing hormone expression in adrenalectomy Wistar rats. Experimental and Therapeutic Medicine. 2016

[31] Faccio L et al. Serum levels of LH, FSH, estradiol and progesterone in female rats experimentally infected by *Trypanosoma evansi*. Experimental Parasitology. 2013;**135**:110-115

**368**

148-161

*Plant Stress Physiology*

**References**

[1] Gangopadhyay N, Hossain MB, Rai DK, Brunton NP. A review of extraction and analysis of bioactives in oat and barley and scope for use of novel food processing technologies. Molecules. 2015;**20**:10884-10909

Nutrition & Food Research. 2009;**53**:

[9] Yeo HB, Yoon HK, Lee HJ, Kang SG, Jung KY, Kim L. Effects of Korean Red Ginseng on cognitive and motor function: A double-blind, randomized, placebo-controlled trial. Journal of Ginseng Research. 2012;**36**(2):190-197

[10] So S, Lee JW, Kim YS, Hyun SH, Han CK. Red ginseng monograph. Journal of Ginseng Research.

[11] Kim J, Cho SY, Kim SH, Kim S, Park CW, Cho D, et al. Ginseng berry and its biological effects as a natural phytochemical. Natural Products Chemistry & Research. 2016;**4**(2):2-5

[12] Rokot NT, Kairupan TS, Cheng KC,

[13] Park J, Song H, Kim SK, Lee MS, Rhee DK, Lee Y. Effects of ginseng on two main sex steroid hormone receptors: Estrogen and androgen receptors. Journal of Ginseng Research.

[14] Kim KH, Lee D, Lee HL, Kim CE, Jung K, Kang KS. Beneficial effects of Panax ginseng for the treatment and prevention of neurodegenerative diseases: Past findings and future directions. Journal of Ginseng Research.

2017;**41**(2):215-221

2018;**42**(3):239-247

[15] Chung IM, Lim JJ, Ahn MS,

Research. 2016;**40**(1):68-75

Jeong HN, An TJ, Kim SH. Comparative phenolic compound profiles and antioxidative activity of the fruit, leaves, and roots of Korean ginseng (*Panax ginseng* Meyer) according to cultivation years. Journal of Ginseng

Runtuwene J, Kapantow NH, Amitani M, et al. A role of ginseng and its constituents in the treatment of central nervous system disorders. Evidence-based Complementary and Alternative Medicine. 2016. ID 2614742

2018;**42**(4):549-561

S330–S375

[2] Youssef MKE, El-fishawy FAE, Ramadan EAE. Nutritional assessment of barley, talbina and their germinated products. Scientific Journal of Crop

Science. 2013;**3**:8-19

[3] Rahman MH, Ali MY. The

[4] Zhu Y, Li T, Fu X, Abbasi AM, Zheng B, Liu RH. Phenolics content, antioxidant and antiproliferative activities of dehulled highland barley (*Hordeum vulgare* L.). Journal of Functional Foods. 2015;**19**:439-450

[5] Idehen E, Tang Y, Sang S. Bioactive phytochemicals in barley. Journal of Food and Drug Analysis. 2017;**25**:

[6] Choi JS et al. Consumption of barley β-glucan ameliorates fatty liver and insulin resistance in mice fed a highfat diet. Molecular Nutrition & Food

Research. 2010;**54**:1004-1013

2011;**141**:1011S–1022S

[7] Jonnalagadda SS et al. Putting the whole grain puzzle together: Health benefits associated with whole grains—Summary of American Society for Nutrition 2010 Satellite Symposium. The Journal of Nutrition.

[8] Schlemmer U, Frølich W, Prieto RM,

significance for humans: Food sources, intake, processing, bioavailability, protective role and analysis. Molecular

Grases F. Phytate in foods and

relationships between thyroid hormones and the brain serotonin (5-Ht) system and mood: Of synergy and significance in the adult brain—A review. Faridpur Medical College Journal. 2015;**9**:98-101

[32] Hosseini M, Dastghaib SS, Rafatpanah H, Hadjzadeh MA-R, Nahrevanian H, Farrokhi I. Nitric oxide contributes to learning and memory deficits observed in hypothyroid rats during neonatal and juvenile growth. Clinics. 2010;**65**:1175-1181

[33] Mancini A, Di Segni C, Raimondo S, Olivieri G, Silvestrini A, Meucci E, et al. Thyroid hormones, oxidative stress, and inflammation. Mediators of Inflammation. 2016:1-12

[34] Gamel TH, Abdel-Aal ESM. Phenolic acids and antioxidant properties of barley wholegrain and pearling fractions. Agricultural and Food Science. 2012;**21**:118-131

[35] Leitao C et al. Fate of polyphenols and antioxidant activity of barley throughout malting and brewing. Journal of Cereal Science. 2012;**55**:318-322

[36] Badrasawi MM, Shahar S, Manaf ZA, Haron H. Effect of Talbinah food consumption on depressive symptoms among elderly individuals in long term care facilities, randomized clinical trial. Clinical Interventions in Aging. 2013;**8**:279-285

[37] Gillies GE, McArthur S. Estrogen actions in the brain and the basis for differential action in men and women: A case for sex-specific medicines. Pharmacological Reviews. 2010;**62**:155-198

[38] Benmansour S, Arroyo LD, Frazer A. Comparison of the antidepressant-like effects of estradiol and that of selective serotonin reuptake inhibitors in middle-aged ovariectomized rats. Frontiers in Aging Neuroscience. 2016;**8**:1-13

[39] Cansev M, Wurtman RJ. Aromatic amino acids in the brain. In: Handbook of Neurochemistry and Molecular Neurobiology. 2007. pp. 59-97

[40] Delisi LE. Handbook of Neurochemistry and Molecular Neurobiology. Handbook of Neurochemistry and Molecular Neurobiology. Schizoprenia. 2009;**27**:107-241

[41] Li B, Zhang S, Li M, Hertz L, Peng L. Serotonin increases ERK1/2 phosphorylation in astrocytes by stimulation of 5-HT2 Band 5-HT2C receptors. Neurochemistry International. 2010;**57**:432-439

[42] Karpinski M, Mattina GF, Steiner M. Effect of gonadal hormones on neurotransmitters implicated in the pathophysiology of obsessivecompulsive disorder: A critical review. Neuroendocrinology. 2017;**105**:1-16

[43] Manna D, Roy G, Mugesh G. Antithyroid drugs and their analogues: Synthesis, structure, and mechanism of action. Accounts of Chemical Research. 2013;**46**(11):2706-2715

[44] Singh RP, Singh A, Sirohi HV, Singh AK, Kaur P, Sharma S, et al. Dual binding mode of antithyroid drug methimazole to mammalian heme peroxidases—Structural determination of the lactoperoxidase-methimazole complex at 1.97 Å resolution. FEBS Open Bio. 2016;**6**:640-650

[45] Hage MP, Azar ST. The link between thyroid function and depression. Journal of Thyroid Research. 2012:1-8 Article ID 590648

[46] McCormack PD, Thomas J, Malik M, Staschen CM. Cold stress, reverse T3 and lymphocyte function. Alaska Medicine. 1998;**40**:55-62

[47] Dai X, Zhou Y, Yu X. Effect of ginseng injection in treating congestive heart failure and its influence on thyroid hormones. Zhongguo Zhong Xi Yi Jie He Za Zhi. 1999;**19**(4):209-211

[48] Kim Y, Choi EH, Doo M, Kim JY, Kim CJ, Kim CT, et al. Anti-stress

**371**

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility…*

[56] Lee S, Rhee DK. Effects of ginseng on stress-related depression, anxiety, and the hypothalamic pituitary adrenal axis. Journal of Ginseng Research.

[57] Kim EH, Kim IH, Ha JA, Choi KT, Pyo S, Rhee DK. Antistress effect of red ginseng in brain cells is mediated by TACE repression via PADI4. Journal of Ginseng Research. 2013;**37**(3):315-323

[58] Kang H, Lim JW, Kim H. Inhibitory effect of Korean red ginseng extract on DNA damage response and apoptosis in *Helicobacter pylori*-infected gastric epithelial cells. Journal of Ginseng

[59] Hassan WA, Aly MS, Rahman TA, Shahat AS. Impact of experimental hypothyroidism on monoamines level in discrete brain regions and other peripheral tissues of young and adult male rats. International Journal of Developmental Neuroscience.

[60] Lee DCW, Lau ASY. Effects of Panax ginseng on tumour necrosis factor-αmediated inflammation: A mini-review.

Molecules. 2011;**16**(4):2802-2816

[61] Hao K, Gong P, Sun SQ, et al. Beneficial estrogen-like effects of ginsenoside Rb1, an active component of Panax ginseng, on neural 5-HT disposition and behavioural tasks in ovariectomized mice. European Journal of Pharmacology. 2011;**659**(1):15-25

[62] Jang D, Lee HLK, Kim KR,

5705232

Won R, Lee SE, Shim I. White ginseng ameliorates depressive behavior and increases hippocampal 5-ht level in the stressed ovariectomized rats. BioMed Research International. 2019. Article ID

2017;**41**(4):589-594

Research. 2018:1-7

2013;**31**(4):225-233

*DOI: http://dx.doi.org/10.5772/intechopen.92359*

effects of ginseng via down-regulation of tyrosine hydroxylase (TH) and dopamine β-hydroxylase (DBH) gene expression in immobilization-stressed rats and PC12 cells. Nutrition Research and Practice. 2010;**4**(4):270-275

[49] Wen L, Jiang X, Sun J, Li X, Li X, Tian L, et al. Cyanidin-3-O-glucoside

progesterone through the protection of mitochondrial function in Pb-exposed rat Leydig cells. Food and Chemical Toxicology. 2018;**112**:427-434

[51] Kuete V. Medicinal Plant Research in Africa: Pharmacology and Chemistry.

activities, biosynthesis and evolution of triterpenoid saponins. Phytochemistry.

[53] Van Kampen JM, Baranowski DB, Shaw CA, Kay DG. Panax ginseng is neuroprotective in a novel progressive model of Parkinson's disease. Experimental Gerontology.

[54] Al-Hazmi MA, Rawi SM, Arafa NM, Wagas A, Montasser AO. The potent effects of ginseng root extract and memantine on cognitive dysfunction in male albino rats. Toxicology and Industrial Health. 2015;**31**(6):494-509

[55] Hussein J, El-Khayat Z, El-Toukhy S, El-Bana M, Medhat D, Morsy S. Panax ginseng regulates brain monoamines in lipopolysaccharide-induced

experimental brain injury. Der Pharma

Chemica. 2016;**8**(6):116-121

promotes the biosynthesis of

[50] Jeyaraj DA, Mani Maran RR, Aruldhas MM, Govindarajulu P. Progesterone induced modulations of serum hormonal profiles in adult male and female rats. Endocrine Research.

2001;**27**(1-2):223-232

1st ed. Elseiver; 2013. p. 890

[52] Augustin JM, Kuzina V, Andersen SB, Bak S. Molecular

2011;**72**(6):435-457

2014;**50**:95-105

*Potential Role of Plants* Hordeum vulgare *L. and* Panax ginseng *L. in Resolving the Fertility… DOI: http://dx.doi.org/10.5772/intechopen.92359*

effects of ginseng via down-regulation of tyrosine hydroxylase (TH) and dopamine β-hydroxylase (DBH) gene expression in immobilization-stressed rats and PC12 cells. Nutrition Research and Practice. 2010;**4**(4):270-275

*Plant Stress Physiology*

[32] Hosseini M, Dastghaib SS, Rafatpanah H, Hadjzadeh MA-R, Nahrevanian H, Farrokhi I. Nitric oxide contributes to learning and memory deficits observed in hypothyroid rats during neonatal and juvenile growth.

Clinics. 2010;**65**:1175-1181

Inflammation. 2016:1-12

[34] Gamel TH, Abdel-Aal ESM. Phenolic acids and antioxidant properties of barley wholegrain and pearling fractions. Agricultural and Food Science. 2012;**21**:118-131

[35] Leitao C et al. Fate of polyphenols

Manaf ZA, Haron H. Effect of Talbinah food consumption on depressive symptoms among elderly individuals in long term care facilities, randomized clinical trial. Clinical Interventions in

[37] Gillies GE, McArthur S. Estrogen actions in the brain and the basis for differential action in men and women: A case for sex-specific medicines. Pharmacological Reviews.

[38] Benmansour S, Arroyo LD, Frazer A. Comparison of the

and that of selective serotonin reuptake inhibitors in middle-aged ovariectomized rats. Frontiers in Aging

Neuroscience. 2016;**8**:1-13

antidepressant-like effects of estradiol

[39] Cansev M, Wurtman RJ. Aromatic amino acids in the brain. In: Handbook of Neurochemistry and Molecular Neurobiology. 2007. pp. 59-97

and antioxidant activity of barley throughout malting and brewing. Journal of Cereal Science.

[36] Badrasawi MM, Shahar S,

Aging. 2013;**8**:279-285

2010;**62**:155-198

2012;**55**:318-322

[33] Mancini A, Di Segni C, Raimondo S, Olivieri G, Silvestrini A, Meucci E, et al. Thyroid hormones, oxidative stress, and inflammation. Mediators of

[40] Delisi LE. Handbook of Neurochemistry and Molecular Neurobiology. Handbook of Neurochemistry and Molecular Neurobiology. Schizoprenia. 2009;**27**:107-241

[41] Li B, Zhang S, Li M, Hertz L, Peng L. Serotonin increases ERK1/2 phosphorylation in astrocytes by stimulation of 5-HT2 Band 5-HT2C receptors. Neurochemistry International. 2010;**57**:432-439

[42] Karpinski M, Mattina GF,

[43] Manna D, Roy G, Mugesh G. Antithyroid drugs and their analogues: Synthesis, structure, and mechanism of action. Accounts of Chemical Research.

[44] Singh RP, Singh A, Sirohi HV, Singh AK, Kaur P, Sharma S, et al. Dual binding mode of antithyroid drug methimazole to mammalian heme peroxidases—Structural determination of the lactoperoxidase-methimazole complex at 1.97 Å resolution. FEBS

[45] Hage MP, Azar ST. The link between thyroid function and depression. Journal of Thyroid Research. 2012:1-8

2013;**46**(11):2706-2715

Open Bio. 2016;**6**:640-650

[46] McCormack PD, Thomas J, Malik M, Staschen CM. Cold stress, reverse T3 and lymphocyte function. Alaska Medicine. 1998;**40**:55-62

[47] Dai X, Zhou Y, Yu X. Effect of ginseng injection in treating congestive heart failure and its influence on thyroid hormones. Zhongguo Zhong Xi Yi Jie He

[48] Kim Y, Choi EH, Doo M, Kim JY, Kim CJ, Kim CT, et al. Anti-stress

Za Zhi. 1999;**19**(4):209-211

Article ID 590648

Steiner M. Effect of gonadal hormones on neurotransmitters implicated in the pathophysiology of obsessivecompulsive disorder: A critical review. Neuroendocrinology. 2017;**105**:1-16

**370**

[49] Wen L, Jiang X, Sun J, Li X, Li X, Tian L, et al. Cyanidin-3-O-glucoside promotes the biosynthesis of progesterone through the protection of mitochondrial function in Pb-exposed rat Leydig cells. Food and Chemical Toxicology. 2018;**112**:427-434

[50] Jeyaraj DA, Mani Maran RR, Aruldhas MM, Govindarajulu P. Progesterone induced modulations of serum hormonal profiles in adult male and female rats. Endocrine Research. 2001;**27**(1-2):223-232

[51] Kuete V. Medicinal Plant Research in Africa: Pharmacology and Chemistry. 1st ed. Elseiver; 2013. p. 890

[52] Augustin JM, Kuzina V, Andersen SB, Bak S. Molecular activities, biosynthesis and evolution of triterpenoid saponins. Phytochemistry. 2011;**72**(6):435-457

[53] Van Kampen JM, Baranowski DB, Shaw CA, Kay DG. Panax ginseng is neuroprotective in a novel progressive model of Parkinson's disease. Experimental Gerontology. 2014;**50**:95-105

[54] Al-Hazmi MA, Rawi SM, Arafa NM, Wagas A, Montasser AO. The potent effects of ginseng root extract and memantine on cognitive dysfunction in male albino rats. Toxicology and Industrial Health. 2015;**31**(6):494-509

[55] Hussein J, El-Khayat Z, El-Toukhy S, El-Bana M, Medhat D, Morsy S. Panax ginseng regulates brain monoamines in lipopolysaccharide-induced experimental brain injury. Der Pharma Chemica. 2016;**8**(6):116-121

[56] Lee S, Rhee DK. Effects of ginseng on stress-related depression, anxiety, and the hypothalamic pituitary adrenal axis. Journal of Ginseng Research. 2017;**41**(4):589-594

[57] Kim EH, Kim IH, Ha JA, Choi KT, Pyo S, Rhee DK. Antistress effect of red ginseng in brain cells is mediated by TACE repression via PADI4. Journal of Ginseng Research. 2013;**37**(3):315-323

[58] Kang H, Lim JW, Kim H. Inhibitory effect of Korean red ginseng extract on DNA damage response and apoptosis in *Helicobacter pylori*-infected gastric epithelial cells. Journal of Ginseng Research. 2018:1-7

[59] Hassan WA, Aly MS, Rahman TA, Shahat AS. Impact of experimental hypothyroidism on monoamines level in discrete brain regions and other peripheral tissues of young and adult male rats. International Journal of Developmental Neuroscience. 2013;**31**(4):225-233

[60] Lee DCW, Lau ASY. Effects of Panax ginseng on tumour necrosis factor-αmediated inflammation: A mini-review. Molecules. 2011;**16**(4):2802-2816

[61] Hao K, Gong P, Sun SQ, et al. Beneficial estrogen-like effects of ginsenoside Rb1, an active component of Panax ginseng, on neural 5-HT disposition and behavioural tasks in ovariectomized mice. European Journal of Pharmacology. 2011;**659**(1):15-25

[62] Jang D, Lee HLK, Kim KR, Won R, Lee SE, Shim I. White ginseng ameliorates depressive behavior and increases hippocampal 5-ht level in the stressed ovariectomized rats. BioMed Research International. 2019. Article ID 5705232

**373**

Section 5

Micoorganisms Mediated

Adaptive Mechanisms to

Abiotic Stresses

## Section 5
