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

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 finegrained perlite [47].

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

**Figure 7.** *Runner bean seedlings treated with Pb grown in viol. (a) general view at development phase. (b) general view after development.*

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.

**Figure 4.**

**Figure 5.**

**Figure 6.**

**8**

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

*Plant Communities and Their Environment*

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

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

microtome by the paraffin method. All plant samples were subjected to various

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

During the examination of the sections, the treatment of plant tissues with dye has caused a problem since the samples contain heavy metals, such as copper and lead, and they affect the physiology of the plant. However, it is known that in permanent preparations, an artificial appearance is obtained by losing some of the chemical content of the plant material and pigments due to the fact that the plant materials pass through a considerable amount of chemical stages. Therefore, it has been stated in some studies that permanent preparation methods are not suitable for some plants [52, 53]. It was also tried to make hand cross sections on artichoke and runner bean samples. The anatomical features of the taxa were evaluated and

Dyes were prepared using different ratios of safranin and fast green, and all of them were tested for staining of heavy metal-treated anatomical specimens in accordance with the literature [50, 51], and examinations were performed on the

Furthermore, objectives of 4, 10, 20, and 40 were used for examination and photographing the anatomical structures of the root and stem of the taxa. Runner bean and artichoke root and stem photographs were taken with 4, 10, and 20 objectives, and the unit of measurement was determined as 100 (μm).

Runner bean seeds were germinated at all concentrations of Cu and Pb. However, there wasn't any germination observed in 640 and 1280 ppm concentrations of Cu in the artichoke seeds, but germination occurred in all

It was determined in the morphological observations of runner bean and artichoke plants subjected to heavy metals that significant differences occurred in

The control group samples of the runner bean and artichoke plants were photographed in order to compare the heavy metal phytotoxic effects on the

treatments to make the sections suitable for microtome removal [47]. These operations were carried out on the samples taken from the abovementioned parts of the plants retained in 70% alcohol. These parts were passed through 80, 90, and 100% alcohol and 2 alcohol/1 xylol, 1 xylol/1 alcohol, 1 alcohol/2 xylol, and 100% xylol solutions, in this order. The paraffin was allowed to penetrate the interior of the samples which were kept in the laboratory drying oven at 60° for 48 hours. Sections of 20, 25, 30, 35, and 40 μm used in the investigations were obtained via samples placed in paraffin blocks. The sections were placed on slides properly by using a hot water bath set at 40 °, and they were fixed onto the slides with adhesive. Sections were cleared off paraffin using 100% xylol, 1 xylol/1 alcohol, absolute alcohol, 95% alcohol, 80% alcohol, 70% alcohol, and purified water, in this order, for 5 minutes each, and then they were stained with safranin and fast green. Samples were kept in pure water, 70% alcohol, 80% alcohol, 95% alcohol, absolute alcohol, 1 xylol: 1 alcohol, and 100% xylol, for 1 minute each, so that water removal from the tissues was completed [49–51]. After removal of all the water, the preparations which were made permanent using Entellan and allowed to dry for 4–5 days at room temperature were examined in general. In this way, the reaction of the samples (with different concentrations of heavy metal in the cells) to

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

dyes and the possible staining status were determined.

interpreted according to Carlquist, Fahn, and Yentür [2, 54, 55].

specimens deemed appropriate.

**4.1 Morphological observations**

concentrations of Pb [39, 47].

different doses of phytotoxic effects of Cu and Pb [47].

**4. Results**

**11**

### **Figure 8.**

*Artichoke seedlings treated with Pb grown in viol. (A) General view. (b) Close-up view of samples treated with 20 ppm Pb.*

### **Figure 9.**

*General view of the control group grown in the climate cabinet, chickpea seedlings treated with Au NPs and C70 SWNTs and pots with late germination.*
