**2. Effects of heavy metals, nanoparticles, and nanotubes on plant phenological development**

The term heavy metal has been used by scientists with various definitions for about 60 years. An element with a density of more than 7 g/cm<sup>3</sup> , in 1987 with a density of more than 4 g/cm<sup>3</sup> , in 1992 with a density greater than 5 g/cm<sup>3</sup> , and in 1995 with a density of 6 g/cm<sup>3</sup> with a metallic property was classified as heavy metal in 1964. Some scientists have classified heavy metals according to their atomic weights, atomic numbers, other chemical properties, and toxic properties. In biological terms, the term heavy metal is generally used for possible contamination of metals and metalloids on the environment and in terms of their toxicity or ecotoxicity [4].

Heavy metals are released into the atmosphere, pedosphere, and hydrosphere every day due to human activities besides natural causes, such as volcanic activities. Flying ashes from the chimneys of cement plants and thermal power plants; the use of heavy metal paint; the smoke emitted by motor vehicles as well as their plasticbased parts such as brake pads, garbage, and waste sludge incineration plants; and the release of industrial wastes, such as pesticides, fertilizers, paper, batteries, products, etc. are among the main causes of heavy metal pollution [5–7].

The discharge of heavy metal-containing particles released from the factory and plant chimneys onto agricultural lands, their dissolution in the soil by rain or irrigation, or the irrigation of agricultural land mixed with industrial wastewater leads to various diseases in crops grown on such lands and damages the agricultural economy [8–10].

Heavy metals have toxic effects for living organisms at certain concentrations. However, certain critical concentrations of some heavy metals are necessary for normal and healthy plant growth. Therefore, heavy metals are classified as essential elements and nonessential elements according to their participation in life processes. Cobalt (Co), copper (Cu), manganese (Mn), molybdenum (Mo), iron (Fe), nickel (Ni), and zinc (Zn) are heavy metals necessary for the growth and vitality of plants and are considered essential elements. Heavy metals such as barium (Ba), cadmium (Cd), mercury (Hg), antimony (Sb), lead (Pb), and chromium (Cr) are not essential for plants and other living organisms and are called nonessential elements [11].

Essential elements are found as a cofactor in many enzyme systems and as a structural component in biological processes in living organisms. For example, copper is an essential element for normal plant growth at certain concentrations. Copper is an essential cofactor for many metalloproteins in plants and plays a role in photosynthetic electron transport, mitochondrial respiration, cell wall metabolisms, and hormone signal transduction pathways [12, 13].

High concentrations of copper (depending on plant species) show toxicity in plants although it is an essential element. Lead is not an essential element and shows toxic properties for plants. The presence of excess copper and lead in the environment negatively affects phenological development in plants [14].

These heavy metals result in lipid peroxidation [15], degradation of cell and thylakoid membrane structure, and a decrease in chlorophyll amount due to the change in the chloroplast structure and thus chlorosis as a result of the oxidative damage they caused [16]. Heavy metals bind to sulfhydryl (-SH) groups of proteins and inhibit enzyme activity [17] and cause oxidative DNA damage [18, 19], chromosomal abnormalities [20], and lack of other essential elements [21–24].

More than 30 base lesions were characterized by DNA exposure to reactive oxygen species [25]. On the DNA, reactive oxygen species can cause

Therefore, it is seen that countries pay particular attention to pollution-related studies and health problems caused by pollution and allocate high amounts of

Heavy metals show toxic effects at certain concentrations for living organisms. However, low concentrations of some heavy metals are essential for normal and healthy plant growth. Furthermore, heavy metals and nanoparticles are causes of concern because they can penetrate into different parts and cells of plants at different rates, and by this way, they enter the food chain and reach the living

There are about 22,000 bryophyte species and 20,000 algae species; however, vascular plants are the dominant plant group in the world with 255,000 species. Land plants, which perform their life cycles completely in the terrestrial environment, are mainly composed of bryophytes and vascular plants. Furthermore, at least a thin film of water is required for fertilization in all taxa except seed plants. Even in the two primitive genera seed plants, cycad and gingko, fertilization is a result of free-swimming spermatozoids released into the liquid medium in the

One of the most important features of vascular plants is the presence of buds at the ends of the trunk and side branches in the gymnosperms and generally in the angiosperms. The bud is an apical meristem coated with protective bud scales. Meristem is the region of cells to which new cells, tissues, and organs are added and has the potential for active cell division and contributes to plant growth. Therefore, despite the limited growth potential in animals, plant growth is limitless due to the presence of apical meristem. However, the development of plant parts, such as

leaves, flowers, and fruits, is limited to their shapes and is genetically

the plant after completing their development.

which the pith and cortex are formed [1, 2].

morphologically and anatomically.

**4**

predetermined [1]. In short, when evaluated from a phenological point of view, plant parts do not show any further growth independent of the time they remain on

Cell development and differentiation take place as the changes occurring in protoplast; for example with the fusion occurring in vacuoles to grow, via structures such as mitochondria, plastids and the golgi body, endoplasmic reticulum, microtubules, and microfilaments in cytoplasm. Cell walls differentiate and increase in thickness due to structural and environmental effects, and they may become permeable. Moreover, the walls may integrate with the lignin, which increases tensile forces. Tissues formed by the differentiation of apical meristem include parenchyma, collenchyma, sclerenchyma, and primary xylem and primary phloem, in

Phenological stages are divided into eight possible principal stages: [1] bud

inflorescence emergence, [5] flowering, [6] fruit development, [7] fruit maturity, and [8] senescence and the beginning of dormancy [3]. Secondary parts and secondary metabolites occur in the plant during the phenological cycle [1]. Genotype and environmental factors are involved in the emergence of seconder metabolites. In this case, based on the amount of soil, water, and air pollution in the environment in which the plant grows, various deteriorations may occur as a result of morphological and physiological changes whose effects on the plant can be seen with the naked eye or observed only through microscopic examinations. In this chapter, general information about heavy metal and nanoparticles is given, and the effects of heavy metals and nanoparticles on the seedlings of runner bean (*Phaseolus coccinea*), chickpea (*Cicer arietinum*), and artichoke (*Cynara scolymus*) species, which are of economic importance, were examined

development, [2] leaf development, [3] shoot/branch development, [4]

resources to deal with the problem.

*Plant Communities and Their Environment*

archegonium chamber [1–3].

beings.

single-nucleobase lesions, single-strand breaks, double-strand breaks, and various oxidative damages such as base connections in the strand [26–28].

Contamination of soils with heavy metals and the accumulation of heavy metals in high concentrations in plants grown here have a genotoxic effect in plants and lead to mutation-like changes in the DNA profile. Therefore, a connection can be established between these changes in the organism and the intensity of pollution in the soil [29].

Sresty and Rao (1999) examined the ultrastructural changes in the nucleolus, nucleus, endoplasmic reticulum, and vacuoles in pea plant stem cells in response to zinc and nickel stress [30].

Zengin and Munzuroğlu (2004) observed the root, stem, and leaf growth in bean (*Phaseolus vulgaris* L.) seedlings exposed to lead and copper stress and examined which tissue was affected more in heavy metal stress [14].

Soudek et al. (2010) exposed flax (*Linum usitatissimum* L.) seeds to different concentrations of lead, nickel, copper, zinc, cadmium, cobalt, arsenic (As), and chromium heavy metals and examined the effects of heavy metal stress on plant germination and root development [31].

Öztürk Çalı and Candan have studied the effects of fungicide on the morphology and viability of pollens of tomato (*Lycopersicon esculentum* Mill.) [32]; the effect of activator application on the anatomy, morphology, and viability of tomato pollen [33]; and influence of activator on meiosis of tomato [34].

Candan and Öztürk Çalı (2015) have observed pollen micromorphology of four taxa of *Anemone coronaria* L. from western Turkey [35]. On the other hand, the authors have compared the pollen morphology and viability of four naturally distributed and commercial varieties of *Anemone coronaria* [36].

Some studies have used various methods based on single-cell gel electrophoresis (comet assay), micronucleus analysis, or cytogenetic analysis in order to investigate the genotoxic effects of pollution on plants.

Steinkellner et al. (1999) treated samples of *Tradescantia* sp. with water from seven regions of Austria where the water was exposed to industrial pollution, and they examined the chromosomal changes in the cells of the root region of the plant by micronucleus analysis. The authors reported negative changes in plant stem cells at the end of the study [37].

Menke et al. exposed the root area of *Arabidopsis thaliana* (L.) to different genotoxic effects. They examined the damage caused by the genotoxic effect in the plant by single-cell gel electrophoresis method and successfully demonstrated the mutagenic effect occurring in stem cell nuclei [38].

The *Comparison of Physiological, Biochemical and Molecular Parameters in Seedlings of Artichoke (Cynara scolymus L.) and Runner Bean (Phaseolus coccineus L.) Seeds Exposed to Lead (Pb) Heavy Metal Stress in the point of Ecological Pollution* was studied with the 2012-057 numbered project supported by Manisa Celal Bayar University [39]. Candan and Batır have presented this scientific important comparison at a conference after the project was completed [40]. On the other hand, Batır has studied on the thesis about determination of the DNA changes in the artichoke seedlings (*Cynara scolymus* L.) subjected to lead and copper stresses [41], and Batır et al. have written an article about that topic [42]. The original PCR photographs of some primers used related runner bean and artichoke samples are given below [39, 41] (**Figures 1**–**6**).

Today, especially due to increasing demand and changing climatic conditions, many studies have been carried out in plant biotechnology regarding more resistant agricultural plants against factors such as drought, salinity, freezing, and heavy metal contamination. Various biological, chemical, and physical methods are

available as regards obtaining biomolecules; for example, nanomaterials with their considerable reactions have much attention in biomass. The basis of this study is to determine the genotoxic effect levels of different stress factors on different plant

**Figure 1.**

**Figure 2.**

**Figure 3.**

**7**

*PCR gel photograph of OP A03 primer used related runner bean samples [39].*

*DOI: http://dx.doi.org/10.5772/intechopen.91904*

*Plant Phenology and An Assessment of the Effects Regarding Heavy Metals, Nanoparticles…*

*PCR gel photograph of OP C05 primer used related runner bean samples [39].*

*PCR gel photograph of OP C20 primer used related runner bean samples [39].*

*Plant Phenology and An Assessment of the Effects Regarding Heavy Metals, Nanoparticles… DOI: http://dx.doi.org/10.5772/intechopen.91904*


### **Figure 1.**

single-nucleobase lesions, single-strand breaks, double-strand breaks, and various

Contamination of soils with heavy metals and the accumulation of heavy metals in high concentrations in plants grown here have a genotoxic effect in plants and lead to mutation-like changes in the DNA profile. Therefore, a connection can be established between these changes in the organism and the intensity of pollution in

Sresty and Rao (1999) examined the ultrastructural changes in the nucleolus, nucleus, endoplasmic reticulum, and vacuoles in pea plant stem cells in response to

Zengin and Munzuroğlu (2004) observed the root, stem, and leaf growth in bean (*Phaseolus vulgaris* L.) seedlings exposed to lead and copper stress and exam-

Soudek et al. (2010) exposed flax (*Linum usitatissimum* L.) seeds to different concentrations of lead, nickel, copper, zinc, cadmium, cobalt, arsenic (As), and chromium heavy metals and examined the effects of heavy metal stress on plant

Öztürk Çalı and Candan have studied the effects of fungicide on the morphology and viability of pollens of tomato (*Lycopersicon esculentum* Mill.) [32]; the effect of activator application on the anatomy, morphology, and viability of tomato pollen

Candan and Öztürk Çalı (2015) have observed pollen micromorphology of four taxa of *Anemone coronaria* L. from western Turkey [35]. On the other hand, the authors have compared the pollen morphology and viability of four naturally dis-

Some studies have used various methods based on single-cell gel electrophoresis (comet assay), micronucleus analysis, or cytogenetic analysis in order to investigate

Steinkellner et al. (1999) treated samples of *Tradescantia* sp. with water from seven regions of Austria where the water was exposed to industrial pollution, and they examined the chromosomal changes in the cells of the root region of the plant by micronucleus analysis. The authors reported negative changes in plant stem cells

Menke et al. exposed the root area of *Arabidopsis thaliana* (L.) to different genotoxic effects. They examined the damage caused by the genotoxic effect in the plant by single-cell gel electrophoresis method and successfully demonstrated the

The *Comparison of Physiological, Biochemical and Molecular Parameters in Seedlings of Artichoke (Cynara scolymus L.) and Runner Bean (Phaseolus coccineus L.) Seeds Exposed to Lead (Pb) Heavy Metal Stress in the point of Ecological Pollution* was studied with the 2012-057 numbered project supported by Manisa Celal Bayar University [39]. Candan and Batır have presented this scientific important comparison at a conference after the project was completed [40]. On the other hand, Batır has studied on the thesis about determination of the DNA changes in the artichoke seedlings (*Cynara scolymus* L.) subjected to lead and copper stresses [41], and Batır et al. have written an article about that topic [42]. The original PCR photographs of some primers used related runner bean and artichoke samples are given below

Today, especially due to increasing demand and changing climatic conditions, many studies have been carried out in plant biotechnology regarding more resistant agricultural plants against factors such as drought, salinity, freezing, and heavy metal contamination. Various biological, chemical, and physical methods are

oxidative damages such as base connections in the strand [26–28].

ined which tissue was affected more in heavy metal stress [14].

[33]; and influence of activator on meiosis of tomato [34].

tributed and commercial varieties of *Anemone coronaria* [36].

the soil [29].

zinc and nickel stress [30].

*Plant Communities and Their Environment*

germination and root development [31].

the genotoxic effects of pollution on plants.

mutagenic effect occurring in stem cell nuclei [38].

at the end of the study [37].

[39, 41] (**Figures 1**–**6**).

**6**

*PCR gel photograph of OP A03 primer used related runner bean samples [39].*

### **Figure 2.** *PCR gel photograph of OP C05 primer used related runner bean samples [39].*

**Figure 3.** *PCR gel photograph of OP C20 primer used related runner bean samples [39].*

available as regards obtaining biomolecules; for example, nanomaterials with their considerable reactions have much attention in biomass. The basis of this study is to determine the genotoxic effect levels of different stress factors on different plant


**3. Materials and methods**

*DOI: http://dx.doi.org/10.5772/intechopen.91904*

Asteraceae family [43–46].

grained perlite [47].

**Figure 7.**

**9**

*after development.*

Fabaceae family to which runner bean and chickpea plants belong and Asteraceae family to which artichoke belongs were selected as the study material. Both of them are families with large numbers of economically important plants in Turkey as well as in the rest of the world. The flowers of the Fabaceae family are zygomorphic in shape and have legume and lomentum fruit. There are many flowers lined up on the flower tray (receptaculum) and head (capitulum) formed by the bracts surrounding these flowers, and there are achene type fruits in the

*Plant Phenology and An Assessment of the Effects Regarding Heavy Metals, Nanoparticles…*

The aim of the present study was to investigate the effects of copper and lead heavy metals on runner bean (*Phaseolus coccineus*) and artichoke (*Cynara scolymus*) seedlings. In addition, the effects of Au nanoparticles and C70 single-walled carbon nanotubes on chickpea (*Cicer arietinum*) seedlings were investigated morphologically. The tolerability of heavy metal and nanoparticle effects by these plants, the cultivation in heavy metal or nanoparticle-contaminated areas, the morphological and anatomical reflections of the changes in genomes, and to which extent the plant's general structure are preserved compared to controls were evaluated in this way.

Runner bean and artichoke seeds were sterilized and then planted for growing. At least 20 seeds were included and observed in the control group and for each heavy metal application. Seeds were planted and germinated in viols with fine-

CuCl2 2H2O and Pb (CH3COO)2 3H2O solutions were applied in concentrations of 20, 40, 80, 160, 240, 320, 640, and 1280 ppm to the runner bean and artichoke seeds planted in groups of 3(Cu 20, Cu 40, Cu 80, Cu 160, Cu 320, Cu 640, Cu 1280 and Pb 20, Pb 40, Pb 80, Pb 160, Pb 320, Pb 640, Pb 1280). This procedure was repeated for 21 days. The seeds of the control group were planted and irrigated with distilled water. As a result, seedlings of the control group and those subjected to Cu-

*Runner bean seedlings treated with Pb grown in viol. (a) general view at development phase. (b) general view*

**3.1 Germination and cultivation of runner bean and artichoke seeds**

Pb heavy metal stress were obtained after 21 days [47] (**Figures 7, 8**).

### **Figure 4.**

*PCR gel photograph of OP C03 primer used related artichoke samples [39, 41].*


**Figure 5.** *PCR gel photograph of OP C05 primer used related artichoke samples [39, 41].*


**Figure 6.** *PCR gel photograph of OP C18 primer used related artichoke samples [39, 41].*

species. Until recently, the investigation of the effects of stress factors as heavy metals, nanoparticules, and nanoparticles on plant phenology remained at cellular, morphological, and anatomical levels.

*Plant Phenology and An Assessment of the Effects Regarding Heavy Metals, Nanoparticles… DOI: http://dx.doi.org/10.5772/intechopen.91904*
