Çiğdem ULUBAŞ, Kadriye ÇAĞLAYAN, Gülşen SERTKAYA

Detection and characterization of plum pox potyvirus (PPV) by DAS-ELISA and RT-PCR/RFLP analysis in Turkiye

Ülkemizin farklı bölgelerinde Plum pox virüs (PPV) (family Potyviridae, genus Potyvirus)'ünün belirlenmesi ve gelişmiş yöntemler ile PPV izolatlarının araştırılması amacı ile, Malatya yöresi dışında kalan ülkemizin önemli yetiştirici bölgelerinde tesadüfi olarak seçilen bahçelerden PPV'ye özgü belirtiler gösteren badem, erik, kayısı, kiraz, nektarin, şeftali, ve vişne ağaçlarından yaprak örnekleri toplanmıştır. Serolojik (DAS-ELISA) çalışmalarda polyclonal antibodiler ve monoclonal antibodiler (MAbs); MabSB (Universal), Mab4DG5 (PPV-D: Dideron), MabAL (PPV-M: Marcus), MabEA24 (PPV-EI Amar) ve MabAC (PPV-C: Cherry) kullanılmıştır. Türkiye'ye ait PPV izolatiarını karakterize etmek için RT-PCR (Reverse transcription-polymerase chain reaction) / RFLP (Restriction fragment length polymorphism) analizi yapılmıştır. RT-PCR/RFLP analizi ile DAS-ELISA ve DASI-ELISA bulgularını destekleyen sonuçlar elde edilmiştir. Toplam 52 örnekten iki adetinin PPV ile enfekteli olduğu ve bu iki kayısı izolatının M straini olduğu belirlenmiştir. Bu çalışma ile Türkiye'de kaysıda PPV-M ırkının varlığına dair önceki araştırma bulguları desteklenmiştir.

Türkiye'de plum pox (Şarka) virüsünün DAS-ELISA ve RT-PCR/RFLP analizi ile saptanması ve tanımlanması

This study was conducted to determine the presence of plum pox virus (PPV) (family Potyviridae, genus Potyvirus) in different regions of Turkey and to characterize PPV isolates by serological and molecular techniques, including ELlSA and PCR/ RFLP. Thus, leaf samples from different stone fruit species (almond, apricot, nectarine, peach, plum and 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, except for Malatya province. Polyclonal antibodies were used to detect the presence of PPV in the plant samples by serological assays (DAS-ELISA). The following monoclonal antibodies (MAbs); Mab SB (Universal), Mab 4DGS (PPV-D: Dideron-specific), Mab AL (PPV-M: Marcus-specific), Mab EA24 (PPV-El Amar-specific) and Mab AC (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 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 of 52 stone fruit samples collected from apricots in Ankara province were infected with the M strain of PPV. This study confirmed the results of the previous work and demonstrated the presence of the PPV-M strain in apricots in Turkey.

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