RESUMO
A full-length infectious cDNA clone of potato virus Y (PVY) was constructed based on an isolate of the PVY(NTN-NW) strain, SYR-II-2-8, which is able to cause potato tuber necrotic ringspot disease (PTNRD). Silent point mutations were introduced to modify sequences similar to the prokaryotic promoter elements detected at the 5' terminus of the P3 coding region of SYR-II-2-8. This, along with modification of the growing conditions of E. coli, led to the successful construction of a stable full-length infectious cDNA clone, named p2-8C3. The biological properties of p2-8C3 were identical to those of SYR-II-2-8, both of which induced PTNRD in potato, veinal necrosis in tobacco, and similar symptoms in the potato cultivars inoculated.
Assuntos
DNA Complementar/genética , Doenças das Plantas/virologia , Potyvirus/genética , Potyvirus/isolamento & purificação , Solanum tuberosum/virologia , Sequência de Aminoácidos , Sequência de Bases , DNA Complementar/metabolismo , DNA Viral/genética , DNA Viral/metabolismo , Dados de Sequência Molecular , Mutação , Potyvirus/classificação , Potyvirus/fisiologia , Nicotiana/virologiaRESUMO
New recombinant strain and genotype of PVY, designated as PVY(NTN-NW) and SYR-III, respectively, shared properties with PVY(NTN) and PVY(N)W has been reported recently. PVY(NTN-NW) predominated in potato fields in Syria and was able to induce potato tuber necrotic ringspot disease (PTNRD). Due to the rapid spread of the recombinant strains of PVY which might be the case of PVY(NTN-NW), a specific and reliable detection method is an essential step to control this strain and minimize its spread. The shared properties of PVY(NTN-NW) and SYR-III with PVY(NTN) and PVY(N)W, however, complicate their identification involving multiple detection methods. Therefore, a multiplex polymerase chain reaction (PCR), that relies on a combination of previously published and newly designed primers was developed for the detection and identification of PVY(NTN-NW) and SYR-III in single or mixed infections with the main PVY strains, PVY(O), PVY(N), PVY(NTN) and PVY(N)W. In addition, the present PCR assay was able to detect the recombination points in the P1 region enabling the differentiation of the variable genotypes of the recombinant strains PVY(NTN-NW), PVY(NTN) and PVY(N)W. The reliability of this PCR assay was confirmed using a significant number of well characterized PVY isolates collected from Syria and Japan including those of PVY(NTN-NW), SYR-III, PVY(O), NA-PVY(N), PVY(N)W and PVY(NTN).
