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Title: IDENTIFICATION AND PARTIAL CHARACTERIZATION OF PRUNUS NECROTIC RINGSPOT VIRUS ON STONE FRUITS IN JORDAN

Author
item SALEM, N - UNIV OF JORDAN AMMAN
item MANSOUR, A - UNIV OF JORDAN AMMAN
item AL-MUSA, A - UNIV OF JORDAN AMMAN
item AL-NSOUR, A - UNIV OF JORDAN AMMAN
item Hammond, Rosemarie

Submitted to: Journal of Plant Pathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/10/2004
Publication Date: 3/1/2004
Citation: Salem, N., Mansour, A., Al-Musa, A., Al-Nsour, A., Hammond, R. 2004. Identification and partial characterization of prunus necrotic ringspot virus on stone fruits in jordan. Journal of Plant Pathology. 86:83-87.

Interpretive Summary: Prunus necrotic ringspot virus (PNRSV) occurs worldwide wherever stone fruits are grown. It is a serious pathogen of many woody species, causing various ringspot diseases in peach, cherry, rose and hops, and mosaic diseases in apple, plum and rose. PNRSV causes serious disease in nurseries and orchards resulting in reduced tree growth and yield. PNRSV is carried on and in pollen grains and is easily transmitted by routine plant propagation methods. In Jordan and the Middle East, little is known of the phytosanitary status of Prunus species with respect to virus diseases. In this study, we have characterized a PNRSV isolate selected from among those identified from stone fruit trees present in various field plot locations in Jordan. Antisera produced against the Jordan isolate, PNRSV-J, was highly specific for PNRSV. Molecular techniques were also developed to detect the virus and will be a valuable tool for use in large scale virus indexing to determine the phytosanitary status of PNRSV in Jordan and for the development of virus-free propagation material for fruit growers.

Technical Abstract: Prunus necrotic ringspot virus (PNRSV, Genus Ilarvirus, Family Bromoviridae) was isolated from stone fruit trees showing virus-like symptoms grown in Jordan. Identification of this virus was based on host range, properties in crude sap, transmissibility, serological tests, and sequence analysis. PNRSV-J has a limited experimental host range. The dilution end point of infectivity was 10-2, the thermal inactivation point was 57oC, and the virus has an in vitro longevity of 16 h at 25oC. PNRSV-J was detected by double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA). PNRSV-J was purified from cucumber leaves harvested 6-8 days after inoculation. The modified purification method gave adequate virus yield for antibody production. Antiserum produced by immunizing a rabbit had a titer of 1024 in direct antigen coating (DAC)-ELISA, with high specificity to PNRSV. An immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) protocol was a useful method for the detection of PNRSV in herbaceous and woody plant tissues. Nucleotide sequence and phylogenetic analysis of RT-PCR products derived from RNA3 of PNRSV-J confirmed its identity as an isolate of PNRSV and revealed that it is a member of Group I (PV32) isolates.