This study was carried out to detect the presence Tomato mosaic virus (ToMV) by biological, serologic and molecular methods in the pepper production areas in Burdur and Denizli provinces. Symptoms related with virus infections on the pepper plants were observed during the surveys. Mechanical inoculation studies, DAS-ELISA (Double-antibody sandwich enzyme linked immunosorbent assay) tests, IC-RT-PCR (Immunocapture Reverse transcription polimerase chain reaction) studies were carried out during the work carried out to define the virus. The plants collected from the field that were suspicious with regards to being infected with the virus were first tested by the ELISA method. It was determined that 10 of the 209 tested samples were infected with ToMV. The samples that gave positive results during ELISA tests were used in the mechanical inoculation studies. Symptoms were observed in the indicator plants within a period of 7-10 days. 20 plant samples that gave negative ToMV values in the ELISA tests but had absorbance values close to positive along with the 10 samples that were tested as positive in the DAS-ELISA were used in the IC-RTPCR method. All of the 30 samples produced expected amplification size of band with 318 bp as observed on agarose gel electrophoresis.

Biological, serological and molecular detection of tomato mosaic virus infecting pepper plants from turkey

espanolEl virus del mosaico del pepino (CMV) es un agente limitante en la produccion y calidad de cultivos agricolas; cuya inoculacion efectiva y diagnostico confiable son esenciales para implementar estrategias de manejo adecuadas y oportunas. Se evaluaron dos protocolos para la inoculacion del CMV en plantas indicadoras (Nicotiana benthamiana y N. tabacum cv. Xanthi) y diferentes metodos de deteccion de este virus. La infeccion de plantas con CMV se logro mediante transmision mecanica e infiltracion. La presencia del virus se confirmo mediante la expresion de sintomas, serologia (ImmunoStrip®), RT-PCR y por secuenciacion. Los resultados confirmaron la efectividad del metodo de transmision mecanica, en el cual, las plantas infectadas presentaron sintomatologia asociada con el virus 15 dias despues de su inoculacion. La prueba serologica permitio detectar la presencia del patogeno solo en las plantas de N. benthamiana, mientras que la prueba de RT-PCR en todas las plantas evaluadas. Los resultados de secuenciacion mostraron altos valores de identidad nucleotidica con las secuencias reportadas en el NCBI para este virus. La transmision mecanica fue el metodo mas efectivo para la inoculacion del CMV y las tecnicas de analisis molecular fueron mas sensibles y confiables para su identificacion, en comparacion con la prueba serologica. EnglishThe cucumber mosaic virus (CMV) is a limiting agent in the production and quality of agricultural crops whose effective inoculation and reliable diagnosis are essential to implement suitable and timely management strategies. Two protocols for CMV inoculation (Nicotiana benthamiana and N. tabacum cv. Xanthi) as indicator plants and different virus detection methods were evaluated. Infection of plants with CMV was achieved by mechanical transmission and infiltration. The presence of the virus was confirmed by expression of symptoms, serology (ImmunoStrip®), RT-PCR and by sequencing. The results confirmed the effectiveness of the mechanical transmis-sion. The infected plants showed symptoms associated with the pathogen 15 days after inoculation. The serological test allowed detecting the presence of the patho-gen only in N. benthamiana plants, while the RT-PCR test in all evaluated plants. The sequencing results showed high nucleotide identity values with the sequences reported in the NBCI for this virus. Mechanical transmission was the most effective method for CMV inoculation, and molecular analysis techniques were more sensi-tive and reliable for CMV identification, compared to serological test.

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Evaluación de métodos para la inoculación y diagnóstico del virus del mosaico del pepino (CMV)

This study was conducted to determine the prevalence of Tomato spotted wilt virus (TSWV) in pepper growing areas in Burdur, Isparta and Denizli provinces of Turkey in 20122013. During surveys TSWV suspected leaf samples were collected. Total of 448 leaf samples were collected from different locations in the region. Presence of TSWV was investigated using Double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA) method. ELISA test results showed that 61 samples were infected with TSWV (13.61%). ELISA positive samples for TSWV were used in the mechanical inoculation studies. Typical symptoms of TSWV were observed on the indicator plants. Molecular studies were carried out on 25 ELISA negative samples and 61 ELISA positive samples (Totally 86 samples). 63 samples were detected positive with RT-PCR (reverse transcriptase polymerase chain reaction) method while 70 samples were found to be infected with TSWV using IC-RT-PCR (immunocapture reverse transcriptase polymerase chain reaction) method. Expected size of band with 276 bp was observed on agarose gel electrophoresis. IC-RT-PCR method was found to be more suitable and sensitive than RT-PCR.

Ali Bal

Papers

Sensitive detection of Tomato spotted wilt virus from pepper plants by DAS-ELISA, RT-PCR and IC-RT-PCR.