NESTED RT-PCR AND IMMUNOCAPTURE RT-PCR FOR DETECTION OF BEET NECROTIC YELLOW VEIN VIRUS ON SUGAR BEET IN LAKE DISTRICT OF TURKEY


Kilic H. C. , Yardimci N.

ROMANIAN AGRICULTURAL RESEARCH, cilt.29, ss.333-337, 2012 (SCI İndekslerine Giren Dergi) identifier identifier

  • Cilt numarası: 29
  • Basım Tarihi: 2012
  • Dergi Adı: ROMANIAN AGRICULTURAL RESEARCH
  • Sayfa Sayıları: ss.333-337

Özet

Rhizomania is a viral disease, caused by beet necrotic yellow vein furovirus (BNYVV) which was detected in Lake District of Turkey sugar beets in 2006-2007. A total of 203 soil samples were collected from 24 different sugar beet production areas of this region. Plants of Rhizomania-susceptible cultivar Kasandra were grown in these soil samples using bait plant techniques. Bait plants root samples were analysed by DAS-ELISA, RT-PCR, Nested Reverse Transcription-Polymerase Chain Reaction (nPCR) and Immunocapture - Reverse Transcription-Polymerase Chain Reaction (IC-RT-PCR) using specific primers. ELISA test results showed that 85 sample were infected with BNYVV. Fifty samples which were negative in the DAS-ELISA tests were used for RT-PCR for the identification of BNYVV. As a result of RT-PCR, 25 samples were determined as positive. RTPCR was carried out using the 016 and 017 specific primers which amplify a 500 base pair (bp) fragment of the read-through region of the coat protein gene of BNYVV. BNYVV detected in bait plant root products of the expected size were amplified by Nested Reverse Transcription-Polymerase Chain Reaction (nRT-PCR) and Immunocapture-Reverse Transcription-Polymerase Chain Reaction (IC-RT-PCR) (326 bp for nPCR). Twenty five samples which were negative in the RT-PCR tests were used for nRT-PCR for the identification of BNYVV. As a result of nRT-PCR 19 samples were determined as positive. At last, 6 samples which were negative in the nRT-PCR tests were used for IC-RT-PCR. As a result of IC-RT-PCR 2 samples were determined as positive. Because ELISA method is simple, quick, efficient and not expensive, its main use can be to analyse BNYVV of the seedlings in the early days. RT-PCR methods are complex and expensive, but they are more sensitive than ELISA.