SEROLOGICAL, BIOLOGICAL AND MOLECULAR DETECTION OF Prunus necrotic ringspot virus ON Rosa damascena Mill. IN TURKEY

Abstract

This study was carried out during 2012 in oil rose (Rosa damascena Mill.) production areas in Burdur prov-ince of Turkey. A total of 102 oil rose leaf samples showing virus-like symptoms were collected from 8 dif-ferent locations. All oil rose leaf samples were tested for Prunus necrotic ringspot virus (PNRSV) using commercially available DAS-ELISA kit with negative and positive control. 16 samples of 102 were found to be infected with PNRSV (16%). DAS-ELISA positive samples were later inoculated onto Cucumis sativus L. cv. Cemre F1, Chenopodium quinoa Wild. and Catharanthus roseus L.G. Don. Inoculation with extracts from PNRSV-positive plants produced chlorotic local lesions on Chenopodium quinoa Wild. The symptoms was not observed on Cucumis sativus L. cv. Cemre F1 and Catharanthus roseus L.G. Don. Total RNA was extracted from leaves of oil rose samples which were positive in the DAS-ELISA and PNRSV- inoculated Chenopodium quinoa Wild. plants. Total RNA was isolated from EZ-10 spin column plant total RNA minipreps kit (Bio Basic, Canada Inc). Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was performed using PNRSV specific primers as described by Rosner (15). The expected frag-ment size of 785 bp was observed after electrophoresis of PCR products in 1% agarose gel. RNA isolated from healthy Chenopodium quinoa Wild. was used as a negative control. No PCR products were amplified from this sample.

Keywords

• Rosa damascena Mill.
• PNRSV
• DAS-ELISA
• RT-PCR