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    Characterization of Plum pox potyvirus (PPV) by DAS-ELISA and RT-PCR/RFLP analysis in Turkey
    (Int Soc Horticultural Science, 2004) Caglayan, K; Sertkaya, G; Ulubas, C; Kölber, M; Krizbai, L
    This study was conducted to determine the presence of Plum pox virus (PPV) (family Potyviridae, genus Potyvirus) in different regions of Turkey and to characterise PPV isolates by serological and molecular techniques including ELISA, PCR/ RFLP. Thus, leaf samples from different stone fruit species, almond, apricot, nectarine, peach, plum, sour and sweet cherry exhibiting various types of symptoms related to PPV were collected from different parts of the canopy from randomly selected orchards in the main stone fruit growing areas in Turkey. Polyclonal antibodies (PAbs) were used to detect the presence of PPV in the plant samples by serological assays (DAS-ELISA). The following monoclonal antibodies (Mabs): Mab5B (Universal), Mab4DG5 (PPV-D: Dideron-specific), MabAL (PPV-M: Marcus-specific), MabEA24 (PPV-El Amar-specific) and MabAC (PPV-C: Cherry-specific) were used to identify the serotyping of PPV isolates. Reverse transcription-polymerase chain reaction (RT-PCR) assays and restriction fragment length polymorphism (RFLP) analysis of RT-PCR products were performed to characterize Turkish PPV isolates. The results of RT-PCR analyses using general primers were in complete agreement with the DAS-ELISA and DASI-ELISA results, showing that 2 apricot samples of 52 stone fruit samples collected from Ankara province were found to be infected with M strain of PPV. This study confirmed the results of the previous work demonstrated the presence of PPV-M strain on apricot in Turkey.
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    Detection of some viruses of stone fruits in mother plant blocks in eastern Mediterranean region of Turkey
    (Int Soc Horticultural Science, 2004) Sertkaya, G; Caglayan, K; Ulubas, C
    Field inspection and sample collection were carried out in mother plant blocks which are also varietal collection belonging to the Ministry of Agriculture and the Universities in Eastern Mediterranean Region of Turkey during early spring and in autumn in 2002. Samples (shoots and leaves) were collected from symptomatic plants in the orchards. 48 samples (3 almond, 9 apricot, 3 nectarine, 10 peach, It plum, 12 sweet cherry) having different symptoms were collected and tested for the presence of Apple chlorotic leaf spot trichovirus (ACLSV), Apple mosaic ilarvirus (ApMV), Cherry leaf roll nepovirus (CLRV), Prune dwarf ilarvirus (PDV), Prunus necrotic ringspot ilarvirus (PNRSV) and Plum pox potyvirus (PPV) by using DAS-ELISA. According to serological assays, the most common virus was PDV. It was followed by PNRSV and ACLSV. The reverse transcription-polymerase chain reaction (RT-PCR) assays were performed for investigation of PDV, PNRSV and PPV in the samples. The following viruses were identified by ELISA and RT-PCR assays: ACLSV (1 sample), CLRV (1 sample), PDV (5 samples), PNRSV (2 samples). Mix infections of ACLSV+PDV (1 sample) and PDV+PNRSV (2 samples) were also found. No PPV-infected samples were detected from the samples collected from mother plant blocks in Eastern Mediterranean Region of Turkey.
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    RT-PCR detection and molecular characterization of Prunus necrotic ringspot virus isolates occurring in Turkey
    (Blackwell Verlag Gmbh, 2004) Ulubas, C; Ertunc, F
    Sour cherry (Prunus cerasus L.), sweet cherry (P. avium L.), peach [P. persica (L.) Batsch], nectarine [P. persica (L.) Batsch var. nucipersica (Suckow) C.K. Schneid], apricot (P. armeniaca L.) and plum (P. domestica L.) trees either known to be infected, or shown to be so during a survey of different stone fruit growing regions of Turkey, were tested for the occurrence of Prunus necrotic ringspot virus (PNRSV) using double antibody sandwich (DAS)-enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR). DAS-ELISA detected virus in only 31 of 486 samples; an additional 20 samples (total infection 51) were found to be infected by PNRSV when RT-PCR was used. The highest virus incidence occurred in nectarine followed by that in peach. In order to identify specific virus strains, the PCR products amplified from the capsid protein (CP) gene of the 20 isolates selected from different provinces were subjected to restriction fragment length polymorphic (RFLP) analysis. The PCR products were digested by eight different restriction endonucleases. Most of the PNRSV isolates were identified as members of group PV96, but a single isolate was a member of group PV32; none of the isolates belonged to group PE5. Phylogenetic analysis divided the virus isolates into three additional major and three minor groups. No relation was found between the geographic distribution and the grouping of the PNRSV isolates based on RFLP.