*Plant Communities and Their Environment*

morphological characteristics of the plants. Moreover, the general appearance and close-up photographs of the samples related to runner bean plant irrigated with Cu 20 ppm and Pb 160 and Pb 1280 ppm concentrations and artichoke plant irrigated with Cu 20 and 40 ppm and Pb 20 and 40 ppm were taken (**Figures 10**–**15**).

Cu and Pb heavy metals caused various phytotoxic effects in cases where the recommended dosage was exceeded or excessive pollution occurred in any other way was determined as the result of the study when the changes in the morphological structures of the plants were examined. Phytotoxicity seen in the morphological structure of the plant emerged as bending, shrinkage, and dark spots on the end of the leaves. On the other hand, while plant root, stem, and leaf lengths increase in low doses, high concentrations (640–280 ppm) cause size reduction and incomplete development [47] (**Figures 10**–**15**).

**Figure 10.**

*(a) Control group of runner bean seedling. (b) runner bean seedlings treated with Cu 20 ppm [47].*

**Figure 11.**

*(a) Runner bean seedling treated with Pb 1280 ppm. (b) Undeveloped runner bean seed treated with Pb 160 ppm [47].*

It was observed that there was an increase in plant root, stem, and leaf sizes in both treatment groups when chickpea plant control group and the plants subjected to Au Nps and C70 SWNTs were compared. Furthermore, it was determined that there were increases in the number of fibrous roots, nodes, and subbranches in both

*Artichoke seedling treated with Cu 40 ppm. (a) General view. (b) Close-up view of the dried leaves [47].*

*(a) Runner bean seedlings treated with Pb 1280 ppm. (b) General view of anomalies in terms of shape,*

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

*(a) Artichoke seedling of control group. (b) Artichoke seedling treated with Cu 20 ppm [47].*

groups (**Figure 16**).

**Figure 14.**

**13**

**Figure 12.**

**Figure 13.**

*chlorosis, and hole [47].*

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

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

### **Figure 12.**

morphological characteristics of the plants. Moreover, the general appearance and close-up photographs of the samples related to runner bean plant irrigated with Cu 20 ppm and Pb 160 and Pb 1280 ppm concentrations and artichoke plant irrigated with Cu 20 and 40 ppm and Pb 20 and 40 ppm were taken (**Figures 10**–**15**). Cu and Pb heavy metals caused various phytotoxic effects in cases where the recommended dosage was exceeded or excessive pollution occurred in any other way was determined as the result of the study when the changes in the morphological structures of the plants were examined. Phytotoxicity seen in the morphological structure of the plant emerged as bending, shrinkage, and dark spots on the end of the leaves. On the other hand, while plant root, stem, and leaf lengths increase in low doses, high concentrations (640–280 ppm) cause size reduction and incomplete

*(a) Control group of runner bean seedling. (b) runner bean seedlings treated with Cu 20 ppm [47].*

*(a) Runner bean seedling treated with Pb 1280 ppm. (b) Undeveloped runner bean seed treated with Pb*

development [47] (**Figures 10**–**15**).

*Plant Communities and Their Environment*

**Figure 10.**

**Figure 11.**

**12**

*160 ppm [47].*

*(a) Runner bean seedlings treated with Pb 1280 ppm. (b) General view of anomalies in terms of shape, chlorosis, and hole [47].*

**Figure 13.** *(a) Artichoke seedling of control group. (b) Artichoke seedling treated with Cu 20 ppm [47].*

**Figure 14.** *Artichoke seedling treated with Cu 40 ppm. (a) General view. (b) Close-up view of the dried leaves [47].*

It was observed that there was an increase in plant root, stem, and leaf sizes in both treatment groups when chickpea plant control group and the plants subjected to Au Nps and C70 SWNTs were compared. Furthermore, it was determined that there were increases in the number of fibrous roots, nodes, and subbranches in both groups (**Figure 16**).

were taken from the samples subjected to Cu 20, 80, and 640 ppm concentrations. Artichoke seedlings of root cross sections treated with Cu 160 ppm and Pb 320 and 640 ppm concentrations and stem cross sections treated with Cu 20 and 160 ppm

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

However, diseases caused by heavy metal stress in the plant, such as chlorosis and necrosis, and epidermal thickening, density of crystallization, increase in hairiness, and thinning in vascular bundles had negative effects on staining in anatomical studies and caused the tissues not to absorb the dye. Furthermore, the presence of heavy metals in the plant content and crystallization prevented the retention of the dye and made staining process difficult. Thus, a large number of experiments with different dyes and dye concentrations have been carried out for the tissue to

*(a) Control group of runner bean seedling root cross section; vascular bundles, cambium, and glandular primordium. (b) Cross section of runner bean seedling treated with Cu 80 ppm; vascular bundles, cambium,*

*(a) Runner bean seedling treated with Cu 640 ppm: (a) general view of root cross section, vascular bundles, endodermis, pericycle, and cambium. (b) Close-up view of root cross section, secretion canals, cambium,*

*(a) Control group of artichoke seedling root cross section. (b) Cross section of artichoke seedling treated with Cu*

*160 ppm root cross-section central cylinder and conduction bundles [47].*

and Pb 1280 ppm concentrations were examined [47] (**Figures 17**–**25**).

absorb the dye into the cell [47].

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

*sclerenchyma, endodermis, pericycle, and casparian strip [47].*

**Figure 17.**

**Figure 18.**

**Figure 19.**

**15**

*endodermis, casparian strip [47].*

**Figure 15.** *Artichoke seedling treated with Pb 40 ppm. (a) general view. (b) close-up view of the dried leaves [47].*

**Figure 16.**

*(a) Chickpea seedling of the control group and the sample treated with Au NPs. (b) Chickpea seedlings treated with C70 SWNTs.*

### **4.2 Anatomical observations**

Micrometer was selected as the unit for measurements taken from root and stem cross sections of the runner bean and artichoke seedlings. Cross sections taken from the roots and stem parts of the plants were considered suitable for evaluation. The roots and stems, epidermis, vascular bundle elements, secretary canals, sclerenchyma, starch sheath, cortex and pith cells, and cambium cells were measured, and the presence and variety of crystals were examined and compared [47].

Photographs of root and stem cross sections of the runner bean and artichoke plants were taken in order to compare the anatomical effects of heavy metal phytotoxic effects on the morphological characteristics of the plants. Runner bean root cross-sectional photographs were taken from root samples subjected to Cu 80 ppm and 640 ppm, and Pb 640 ppm concentrations and stem cross-section photographs

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

were taken from the samples subjected to Cu 20, 80, and 640 ppm concentrations. Artichoke seedlings of root cross sections treated with Cu 160 ppm and Pb 320 and 640 ppm concentrations and stem cross sections treated with Cu 20 and 160 ppm and Pb 1280 ppm concentrations were examined [47] (**Figures 17**–**25**).

However, diseases caused by heavy metal stress in the plant, such as chlorosis and necrosis, and epidermal thickening, density of crystallization, increase in hairiness, and thinning in vascular bundles had negative effects on staining in anatomical studies and caused the tissues not to absorb the dye. Furthermore, the presence of heavy metals in the plant content and crystallization prevented the retention of the dye and made staining process difficult. Thus, a large number of experiments with different dyes and dye concentrations have been carried out for the tissue to absorb the dye into the cell [47].

### **Figure 17.**

*(a) Control group of runner bean seedling root cross section; vascular bundles, cambium, and glandular primordium. (b) Cross section of runner bean seedling treated with Cu 80 ppm; vascular bundles, cambium, sclerenchyma, endodermis, pericycle, and casparian strip [47].*

### **Figure 18.**

**4.2 Anatomical observations**

**Figure 15.**

*Plant Communities and Their Environment*

**Figure 16.**

**14**

*with C70 SWNTs.*

Micrometer was selected as the unit for measurements taken from root and stem cross sections of the runner bean and artichoke seedlings. Cross sections taken from the roots and stem parts of the plants were considered suitable for evaluation. The roots and stems, epidermis, vascular bundle elements, secretary canals, sclerenchyma, starch sheath, cortex and pith cells, and cambium cells were measured, and

*(a) Chickpea seedling of the control group and the sample treated with Au NPs. (b) Chickpea seedlings treated*

*Artichoke seedling treated with Pb 40 ppm. (a) general view. (b) close-up view of the dried leaves [47].*

Photographs of root and stem cross sections of the runner bean and artichoke plants were taken in order to compare the anatomical effects of heavy metal phytotoxic effects on the morphological characteristics of the plants. Runner bean root cross-sectional photographs were taken from root samples subjected to Cu 80 ppm and 640 ppm, and Pb 640 ppm concentrations and stem cross-section photographs

the presence and variety of crystals were examined and compared [47].

*(a) Runner bean seedling treated with Cu 640 ppm: (a) general view of root cross section, vascular bundles, endodermis, pericycle, and cambium. (b) Close-up view of root cross section, secretion canals, cambium, endodermis, casparian strip [47].*

**Figure 19.**

*(a) Control group of artichoke seedling root cross section. (b) Cross section of artichoke seedling treated with Cu 160 ppm root cross-section central cylinder and conduction bundles [47].*

### **Figure 20.**

*(a) Close-up view of artichoke seedling root treated with Pb 320 ppm: Central cylinder, endodermis, pericycle, and crystals. (b) Close-up view of root artichoke seedling treated with Pb 640 ppm: Vascular bundles and crystals [47].*

### **Figure 21.**

*(a) General view of control group runner bean seedling stem cross section; close-up view of epidermis, cortex bundles, and secretion canals. (b) Close-up view of runner bean seedling stem treated with Cu 20 ppm: Xylem, phloem, secretion canals, and starch scabbard [47].*

### **Figure 22.**

*(a) General view of runner bean seedling stem treated with Cu 80 ppm: Vascular bundles, secretion canals, and crystals. (b) Close-up view of runner bean seedling stem treated with Cu 640 ppm: Secretion canals and crystals [47].*

**5. Discussion**

**Figure 25.**

**17**

**Figure 24.**

*view of artichoke seedling stem treated with Cu 160 ppm.*

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

Heavy metal pollution in soil and water is one of the most important environmental problems in industrialized countries. Various heavy metals such as cad-

*(a)general view of artichoke seedling stem treated with Cu 20 ppm stem: Vascular bundles and (b) general*

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

High-structured plants are equipped with advanced features that allow them to adapt to changes in nature, one of which is the retention of metals in the roots [60]. Retention and deposition of metals in roots have more negative effect on the root area and seed germination than stem and leaf growth. Zengin and Munzuroğlu (2004) reported that the most negative effect was in the root area of the bean (*Phaseolus vulgaris*) seedlings exposed to increasing concentration of lead and copper solutions; stem and leaf growth was negatively affected; however, they stated

mium, lead, copper, mercury, and chromium from various industrial

*(b) close up view of artichoke seedling stem treated with Pb 1280 ppm cortex and crystals.*

that the most negative effect was in the root area of the seedlings [14].

living things by forming very complex structures [59].

establishments such as leather, paint, fertilizer, textile, cement, and chemical industry are released onto soil and aquatic environments and cause environmental pollution [56–58]. Since most of the heavy metals do not undergo biodegradation in the environment, they can easily accumulate and increase their toxic effects on

*(a) Close up view of artichoke seedling stem treated with Cu 160 ppm:Xylem, phloem, and crystals and*

### **Figure 23.**

*(a) Close-up view of runner bean seedling stem treated with Pb 40 ppm: Xylem, phloem, secretion canals, and crystals. (b) General view of runner bean seedling stem treated with Pb 640 ppm: Vascular bundle, secretion canals [47].*

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

### **Figure 24.**

**Figure 21.**

**Figure 20.**

**Figure 22.**

*crystals [47].*

**Figure 23.**

**16**

*phloem, secretion canals, and starch scabbard [47].*

*Plant Communities and Their Environment*

*(a) General view of control group runner bean seedling stem cross section; close-up view of epidermis, cortex bundles, and secretion canals. (b) Close-up view of runner bean seedling stem treated with Cu 20 ppm: Xylem,*

*(a) Close-up view of artichoke seedling root treated with Pb 320 ppm: Central cylinder, endodermis, pericycle, and crystals. (b) Close-up view of root artichoke seedling treated with Pb 640 ppm: Vascular bundles and crystals [47].*

*(a) General view of runner bean seedling stem treated with Cu 80 ppm: Vascular bundles, secretion canals, and crystals. (b) Close-up view of runner bean seedling stem treated with Cu 640 ppm: Secretion canals and*

*(a) Close-up view of runner bean seedling stem treated with Pb 40 ppm: Xylem, phloem, secretion canals, and crystals. (b) General view of runner bean seedling stem treated with Pb 640 ppm: Vascular bundle, secretion canals [47].*

*(a)general view of artichoke seedling stem treated with Cu 20 ppm stem: Vascular bundles and (b) general view of artichoke seedling stem treated with Cu 160 ppm.*

### **Figure 25.**

*(a) Close up view of artichoke seedling stem treated with Cu 160 ppm:Xylem, phloem, and crystals and (b) close up view of artichoke seedling stem treated with Pb 1280 ppm cortex and crystals.*

## **5. Discussion**

Heavy metal pollution in soil and water is one of the most important environmental problems in industrialized countries. Various heavy metals such as cadmium, lead, copper, mercury, and chromium from various industrial establishments such as leather, paint, fertilizer, textile, cement, and chemical industry are released onto soil and aquatic environments and cause environmental pollution [56–58]. Since most of the heavy metals do not undergo biodegradation in the environment, they can easily accumulate and increase their toxic effects on living things by forming very complex structures [59].

High-structured plants are equipped with advanced features that allow them to adapt to changes in nature, one of which is the retention of metals in the roots [60]. Retention and deposition of metals in roots have more negative effect on the root area and seed germination than stem and leaf growth. Zengin and Munzuroğlu (2004) reported that the most negative effect was in the root area of the bean (*Phaseolus vulgaris*) seedlings exposed to increasing concentration of lead and copper solutions; stem and leaf growth was negatively affected; however, they stated that the most negative effect was in the root area of the seedlings [14].

Soudek et al. (2010) treated linen (*Linum usitatissimum* L.) seeds with different concentrations of lead, nickel, copper, zinc, cadmium, cobalt, arsenic, and chromium heavy metals and reported that heavy metal stress had negative effects on the number of germinating seeds and seedling root development. The negative effect of heavy metal stress on root length in plants may result from the division of cells in the root region or the prolongation of the cell cycle [31]. The root, stem, and leaf structures of the runner bean and artichoke seedlings grown in high Cu and Pb concentrations (160, 320, 640, 1280 ppm) examined in this study were degraded as a result of oxidative damage. Therefore, the epidermal cells forming the surface of these parts were damaged, which negatively affected root, stem, and leaf growth. However, it also caused adverse conditions such as dryness, shrinkage, and necrosis in the leaves although Cu and Pb heavy metals applied at low concentrations generally stimulated growth and increase the number of leaves in the plant [39, 47]. Furthermore, browning caused by heavy metal stress was observed in the roots of the runner bean and artichoke seedlings in which high concentrations of heavy metals were applied. This color change occurs with the increase in the amount of suberin in stem cells. Therefore, suberin stem cells will limit the uptake of water, and plant growth inhibition occurs [61, 62].

crystallization is particularly important and has shown similar responses in both heavy metal treatments. The differences varied in terms of crystal density, location,

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

As a result, various anatomical differences determined in this study regarding the characteristic features, such as root and stem vascular bundles thicknesses, cell size and fragmentation in the pith region, formation or thickening of cambium and sclerenchyma, shape and size of secretary canals, differences in cortex cell sizes, the sizes of the vascular bundles, the formation of crystals and deposits in the phloem layer according to the heavy metal concentration applied, the number of epidermis cells per unit area, and the epidermal wall shapes, can be used to reveal the phyto-

The morphology of roots and shoots is extremely important for the growth and

development of all plants, and each factor that changes their morphology has

It is the fact that the application of some materials which are not used consciously or at the recommended dosage and which contain heavy metals in order for the crop to be attractive for the consumers actually yields negative results. Therefore, this study is important about examining the extent of heavy metal phytotoxic effects related determining them on plants phenological development in the point of morphologically and anatomically changes. The information given in this study is valuable as it presents the negative molecular effect of heavy metal pollution on the plant in terms of morphological and anatomical aspects. Furthermore, this study will guide the researchers on the effects of environmental pollution in relation with the phenological development of economic plants and hence on

Heavy metal and nanoparticule-nanotube-induced plants must be evaluated in the point of biochemistry and examined via scanning electron microscope (SEM) and transmission electron microscope (TEM) regarding their root, stem, and leaf structure and apical tip, leaf-bud primordiums, and provascular tissue in detailed ways for interdisciplinary studies according to plants' phenological development progress.

I would like to thank Manisa Celal Bayar University (Turkey) Scientific Research Projects Coordination Unit (BAP) for supporting most of this study as part of the

I am also thankful to Dr. Qi Lu, a member of the Department of Physics and Engineering, and Dr. Gulnihal Ozbay, a member of the Department of Agriculture and Natural Resources at Delaware State University (USA), for the support of some

on the development of chickpea have been observed (**Figure 16**).

On the other hand, Candan and Lu (2017) have shown that there are more differences on the pea green (*Pisum sativum*) anatomy under the effects of C70 nanomaterial [70]. Candan and Markushin have studied about spectroscopic study of the gold nanoparticles (Au NPs) distribution in leaf, stem, and root of the pea green plant [71]. In this chapter, the effects of Au nanoparticles and C70 nanotubes on the morphology of roots, leaves, and stems are investigated, and positive results

and shapes (**Figures 17**–**25**) [47].

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

toxic effects of Cu and Pb heavy metals.

positive/negative effects [69].

**6. Conclusion**

human health.

**Acknowledgements**

**19**

projects numbered 2012-057 and 2014-120.

laboratory studies related to nanoparticles and nanotubes.

The defense mechanisms developed by plants against heavy metal stress may vary in the level of family, genus, species, subspecies, and variety [54–57]. The defense mechanisms that allow plants to be tolerant to heavy metals have not yet been fully understood. However, the mechanisms of tolerance include vacuolar phenomena [58], enzymatic and nonenzymatic antioxidant systems [59, 60], metal-binding ligands such as metallothionein [61], and alternative oxidase pathways [62].

Although copper is an essential element, it is more toxic when it is present in high doses compared to cadmium, a nonessential element. This is explained by the direct influence of copper on the formation of reactive oxygen species [superoxide radical (O2˙), hydrogen peroxide (H2O2), hydroxyl radical (OH)] because it is a trace element. Since copper and iron transition metals are involved in oxidoreduction reactions, they act as catalysts that accelerate the formation of reactive oxygen species [63–66]. As in previous studies, it was found that artichoke seedlings are negatively affected mostly by copper in molecular terms [39, 41, 67, 68]. The highest negative effect was observed in groups subjected to copper solutions in terms of root length, root dry weight, total soluble protein amount, and genomic mold stability [39–42] (**Figures 17**–**22**).

Reactions to heavy metal stress in this study emerged as different anatomical results in both plants. However, it can be determined that the response of coppertreated samples was not always greater than that of the lead-treated samples even though they responded differently to different concentrations in terms of anatomical results. Previously the effect rate observed at the molecular level in Cu-treated samples has been found to be higher than the effect rate in Pb-treated samples in both plants [39–42]. However, it was not the case for the present anatomical study because the roots and stems of the plants were examined in terms of many parameters, so heavy metals did not show the same effect [47] (**Figures 17**–**25**).

It was observed that seedlings belonging to runner bean species showed high tolerance against lead and copper stress. The genome of the plants was preserved at 94–95%, and no significant reduction in total soluble protein was observed especially at 1280 ppm concentration of lead and copper solutions. This situation has led to the conclusion that runner bean species has a strong defense mechanism against heavy metal contamination [40–42, 49]. In this anatomically based study, it would be wrong to say that one heavy metal is always superior to another in terms of its effects because plant parts were examined anatomically in terms of many parameters and similar results were not observed in all. The reaction of the plants as

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

crystallization is particularly important and has shown similar responses in both heavy metal treatments. The differences varied in terms of crystal density, location, and shapes (**Figures 17**–**25**) [47].

As a result, various anatomical differences determined in this study regarding the characteristic features, such as root and stem vascular bundles thicknesses, cell size and fragmentation in the pith region, formation or thickening of cambium and sclerenchyma, shape and size of secretary canals, differences in cortex cell sizes, the sizes of the vascular bundles, the formation of crystals and deposits in the phloem layer according to the heavy metal concentration applied, the number of epidermis cells per unit area, and the epidermal wall shapes, can be used to reveal the phytotoxic effects of Cu and Pb heavy metals.

The morphology of roots and shoots is extremely important for the growth and development of all plants, and each factor that changes their morphology has positive/negative effects [69].

On the other hand, Candan and Lu (2017) have shown that there are more differences on the pea green (*Pisum sativum*) anatomy under the effects of C70 nanomaterial [70]. Candan and Markushin have studied about spectroscopic study of the gold nanoparticles (Au NPs) distribution in leaf, stem, and root of the pea green plant [71]. In this chapter, the effects of Au nanoparticles and C70 nanotubes on the morphology of roots, leaves, and stems are investigated, and positive results on the development of chickpea have been observed (**Figure 16**).
