Cytological and Molecular Studies of an Egyptian Isolate of Carnation Vein Mottle Potyvirus
Cytological and Molecular Studies of an Egyptian Isolate of Carnation Vein Mottle Potyvirus
*Amal Abou El-Ela A., M. A. Amer and Eman A. H. Khatab,
ABSTRACT
Carnation vein mottle virus CarVMV was isolated from infected carnation plants and identified by host-range, serological detection and maintained on Carnation (Dianthus caryophyllus L.) and / or Gompherena globosa. Morphological studies of CarVMV were conducted by light and electron microscopy. Light and electron microscopy revealed amorphous cytoplasmic inclusions in infected leaf cells. In some cases, however, inclusions have a characteristic shape, spindle, circular or sledge-like. Pinwheel inclusions characteristic of Potyvirus which include CarVMV were also observed. The nuclei were variable in size and shape, sometimes complete destruction of the nucleolus could be observed. The virus infection can cause many cytological changes in the chloroplasts and mitochondria. A combined assay of reverse transcription and polymerase chain reaction utilizing degenerate primers derived from conserved regions in the genome of potyviruses was designed to amplify a 335 bp cDNA fragment from infected plant using degenerate oligonucleotide primers specific for detection of Potyvirus group. Amplification of total RNA extracted from infected carnation suggesting the presence of one Potyvirus in the tested carnation plant. Nucleic acid spot hybridization assay also was used to confirm identification of the present virus as one of the Potyvirus group using DIG labeled general cDNA probe for detection of potyviruses group, It showed high levels of hybridization signal, whereas no RT-PCR and hybridization were observed with uninfected carnation tissues.
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July 2006
Vol. 3, Iss. 1
