Molecular Analysis of Cucumber Mosaic Cucumovirus Symptoms Development on Squash Plants

Abstract: A wide range of severe symptoms were appeared on inoculated squash plants (Cucmus pepo cv. ElSkandrani) with CMV under greenhouse condition. One of the first signs of systemic CMV infection, is vein clearing in the youngest leaves (about 7 days), the veins become translucent and leave produced subsequently showed a mosaic (about 10 days) severe mosaic and mottling (about 15 days). Then changed in leaves growth form i.e. little, malformation and no-Lamina giving shoestring the so-called fern leaf (about 20-25 days). The virus was transferred from each symptom to squash and chenopodium amaranticolor plants by sap mechanical inoculation then conformed by DBIA-assay. The change in chlorophyll contents to ensure that all these symptoms resulted from CMV-s EG. SDS-PAGE, peroxidase isozyme separation and RAPD-PCR to molecular analyze of S-CMV-EG symptoms development on squash plant leaves. SDS-PAGE of protein separation showed variability protein pattern and contents of healthy and S-CMV symptoms infected squash leaves (m-mosaic, S-mosaic, crinkling and malformation) with 24, 26, 28, 25, 20 and 22, 21, 20, 22, 16 soluble and unsoluble polypeptides respectively. As well as. DISC-PAGE isozyme showed 6, 7, 8 and 8 peroxidase isozymes respectively. RAPD-analysis revealed DNA polymorphic among CMV-symptom development on squash plants. RAPD analysis using two random primers revealed 8 polymorphic of total 15 amplified fragments with 53% under CMV infection. Crinkle symptoms revealed the highest number with 18 markers followed by S-mosaic 17 and malformation 16 and mild mosaic with 15 bands