DETECTION AND PHYLOGENETIC ANALYSIS OF PRUNUS NECROTIC RINGSPOT VIRUS ISOLATES FROM STONE FRUITS IN IRAN

N. Sokhandan-Bashir, Z. Kashiha, D. Koolivand, O. Eini

Abstract


Symptomatic stone fruit trees suspected to be infected with Prunus necrotic ringspot virus (PNRSV) were sampled in the west and northwest parts of Iran and tested by enzyme-linked immunosorbent assay (ELISA). Then, total RNA from 123 samples from the infected orchards were tested by reverse transcription polymerase chain reaction (RT-PCR) with the ilarvirus universal primer set, Ilar1/ Ilar2. An expected 206 bp DNA fragment was amplified from 30 samples and when these positive samples were subjected to RT-PCR with a pair of specific primers, VP81/ VP103, the coat protein (CP) gene was amplified from 21 out of the 30 samples. Sixteen of the amplified CP fragments were purified, sequenced and the sequence data were compared among the newly sequenced isolates, and with the CP of previously reported isolates. The new isolates were 97-100% and 96-100% homologous at the nucleotide (nt) and deduced amino acid (aa) levels, respectively. However, there were 83-99% (nt) and 74-99% (aa) homologies between these and previously reported isolates. Neighbor-Joining phylogenetic trees were generated on the basis of the nt or aa data of the CP showing that the newly-sequenced isolates were placed within the previously defined PNRSV group known as PV-96-II, which comprises isolates from diverse geographical regions such as Poland, Uruguay, China, Chile, Montenegro, and the USA. This is the first report of detection and 2 genetic analysis of PNRSV isolates from stone fruits in Iran. Also, PNRSV in nectarine is being reported for the first time from the country.

Keywords


Coat protein; ELISA; Ilarvirus; phylogenetic; PNRSV; RT-PCR

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DOI: http://dx.doi.org/10.4454/jpp.v99i3.3986

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