Obtaining of infectious clones corresponding to the genomes of RNA viruses has greatly enhanced investigations.The different strategies employed to construct infectious clones from plant RNA virus and parameters affecting infectivity of such clones have been described.The system represent a powerful tool fo studying plant virus replication, short and long distance virus movement and the virus - host interactions.It may serve as autonomously replicating vectors for the expression of foreign genes in plants.
Binary plasmids carrying different fragments of the coat protein gene of potato leafroll luteovirus were constructed for transformation of commercial potato cultivar Bzura. Several transgenic plant lines were obtained and characterised. The resistance of transgenic plants to the virus was tested by inoculation with aphids or grafting. Selected transformants expressing viral RNA were resistant to virus challenge by viruliferous aphids. Expression of the antisense RNA prevented virus infection even after grafting with scions from infected plants.
In recent years a new subgroup of necrotic potato virus Y (PVY) isolates has spread in the Polish fields. To counter this infection, the PVY resistance was introduced into selected potato and tobacco cultivars. Plants were transformed by agroinfection with pROK2 derived binary plasmids, carrying an appropriate fragment of the PVY genome (patent applied for). Kanamycin resistant plants were screened by PCR for the presence of PVY cDNA inserts, positive transformants were tested for virus resistance by inoculation with sap from infected plants, followed by observations of disease symptoms and virus accumulation analysis. Several resistant potato and tobacco clones were identified and their phenotypes were preliminarily characterised.
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