**4.1 Isolation and diagnosis of cucumber CMV mosaic virus**

#### *4.1.1 Symptoms*

The symptoms appeared in mosaic on cucumber plants inoculated with CMV virus after a week of inoculation and over time the symptoms turned into deformation of the leaves and their edges bending down and twisting and stunting of the plant, as well as shortening of the necks of the leaves in addition to the lack of fruit numbers, deformation and dropping of flowers, and with the progression of the infection Areas wherein turns into yellow areas and then into dead areas (Necrosis), and no symptoms of infection with the virus appeared on healthy plants protected from infection.

#### *4.1.2 CMV molecular diagnostics*

#### *4.1.2.1 Results of RNA isolation*

Isolation of DNA from the leaves of cucumber plants infected with the virus, and an appropriate percentage of RNA was obtained, as the Kit (ready-made kit) method was used to obtain RNA in appropriate quantities ranging between (12.696– 7.420) nanograms per microliter and the purity was (1.71) This result came after repeated attempts because of what is known that the process of isolating RNA is more difficult than DNA because it is sensitive to cracking by the RNase enzymes (Hassan, 2004).

#### *4.1.2.2 Results of PCR reactions*


Forward 2 and Forward 3, they did not show any inflation as in **Figure 7**. This result of the molecular diagnostic test is similar to the findings of Shomaila Igbal et al. [103] in Rawalpindi region/ State of Pakistan, while the rest of the primers did not match, and this indicates a great convergence between the aforementioned isolation in Al-Huwaish/ Samarra district/Salahuddin with the Pakistani isolation. And a difference from the isolation in the regions (Punjab, Sind, Northwest Frontier

*The result of molecular detection of the local isolate of CMV virus for the first and second initiator. M: represents the volumetric index; Forward 1,2: represents the first and second primers where no bundles appeared; H: stands for cucumber. B: represents the pepper. L: represents tomato; T: symbolizes Al-Taroozi plant.*

*A result of molecular detection of the local isolation of CMV virus for the third and fourth initiator. M: represents the volumetric index. Forward 4: H: represents the initiator of 207 bp for the fourth primer. B, L,T:*

*The Activity of New Bio-Agent to Control* Cucumovirus Cucumber Mosaic Virus *(CMV)*

*Forward 3 represents the third primer. No package appeared.*

*DOI: http://dx.doi.org/10.5772/intechopen.96587*

Province, Islamabad and Baluchistan).

**Figure 6.**

**Figure 7.**

**161**

When using the program mentioned in paragraph (3.3.1.5) the straight segments responsible for forming the virus' protein envelope were enlarged with the fourth primer (Forward 4: H), where the reaction showed very clear Bands with size (207 bp), and as They are shown in **Figure 6**. As for the primers Forward 1,

*The Activity of New Bio-Agent to Control* Cucumovirus Cucumber Mosaic Virus *(CMV) DOI: http://dx.doi.org/10.5772/intechopen.96587*

#### **Figure 6.**

*A result of molecular detection of the local isolation of CMV virus for the third and fourth initiator. M: represents the volumetric index. Forward 4: H: represents the initiator of 207 bp for the fourth primer. B, L,T: Forward 3 represents the third primer. No package appeared.*

#### **Figure 7.**

*The result of molecular detection of the local isolate of CMV virus for the first and second initiator. M: represents the volumetric index; Forward 1,2: represents the first and second primers where no bundles appeared; H: stands for cucumber. B: represents the pepper. L: represents tomato; T: symbolizes Al-Taroozi plant.*

Forward 2 and Forward 3, they did not show any inflation as in **Figure 7**. This result of the molecular diagnostic test is similar to the findings of Shomaila Igbal et al. [103] in Rawalpindi region/ State of Pakistan, while the rest of the primers did not match, and this indicates a great convergence between the aforementioned isolation in Al-Huwaish/ Samarra district/Salahuddin with the Pakistani isolation. And a difference from the isolation in the regions (Punjab, Sind, Northwest Frontier Province, Islamabad and Baluchistan).
