See **Figure 19**.

The microscopy result after culturing the callus in an M and S liquid media of () and (+) Kinetin shows that the cells from the media with Kinetin developed more giant cells and several cells after 1 week of cultivation, while the cells in the media with Kinetin developed smaller and multiple cells. However, after 4-weeks, the cells from the media with Kinetin developed more giant cells in more significant numbers. In contrast, the cells from the media without Kinetin incurred contamination before the fourth week, and this result agrees with the findings of Usha et al. and Debnath [99, 103], which explains the necessity of administering a 14-day dark treatment for callus formation and the shoot regeneration of plants. The variation between the cells in media with Kinetin and media without Kinetin could be due to the addition of Kinetin, a growth regulator. It regulates the growth processes of cells by controlling cell division and cell differentiation affecting the plant's formation and regeneration process. Also, this correlates with the results of Usha et al. [99] that envaulted the

#### **Figure 19.**

*(i) Chicory cells in liquid media (+kinetin) no contamination. (ii) chicory cells in liquid media ( kinetin) no contamination.c.) cells after 4 weeks of transferring into new cell suspension culture (liquid culture media) See Figures 20 and 21.*

#### **Figure 20.**

*(i) chicory cells in liquid media without () kinetin presence of contamination.*

**Figure 21.** *(i) chicory cells in liquid media with (+) kinetin absence of contamination.*

effect of plant regulators on plant cells. In addition, external factors, such as light intensity and temperature, could have affected the slow development of cells in the media with Kinetin. Since temperature and light play essential roles in plant growth, this agrees with the findings of Yildiz [57] elaborated on the influence of control equipment causing the culture room to be overheated resulting from temperature

changes, which causes stress to plants enabling very low or no success in the growth rate.

#### **3.6 Wet and dry matter determination**

Two Factors – (I) Liquid culture media with (+) Kinetin and (II) Liquid culture media without () Kinetin were used to determine the outcome for wet and dry weight analysis of callus of chicory culture in liquid media.

#### *3.6.1 Liquid culture media with kinetin (+ kinetin)*

The result, as shown in **Figure 22**, can be deduced that the wet weight of samples A (0.81714), B (0.98178), and C (0.92866) had a close range in the values obtained for the wet weight and that of the dry weight for all samples showed relatively very low variation during drying hence, the dry weight between the samples is approximately the same. The sample with the lowest weight after drying was sample A (0.16974), next to it was sample C (0.17406), then B (0.17978). Thus, the upsurge in dry weight was due to cell splitting 14 and new material synthesis (see **Table 4**) [104, 105].
