Domates Bitkisinde Pospiviroid Cinsi Viroid Etmenlerinin Varlığının Araştırılması ve Moleküler Karakterizasyonu*

Çalışma domates bitkisinde Pospiviroid cinsi viroidlerin varlığının araştırılması için yürütülmüştür. 2014-2015 arasında, 400 adet domates tohum, fide ile yaprak örneği toplanmış ve total nükleik asit ekstraktları elde edilmiştir. Pospi1-FW/RE ve Vid-FW/RE primerleri ile yürütülen RT-PCR çalışmalarında, 398 adet domates örneğinin bir viroidle bulaşık olmadığı görülmüştür. ‘’Çeri‘’ çeşidi iki domates örneğinin ise Citrus exocortis viroid (CEVd) etmeni ile bulaşık olduğu, CEVd-FW/RE primer çifti ile belirlenmiştir. Agaroz jel elektroforezi çalışmalarında bu örneklere ait 371 bp’lik DNA bantları görüntülenmiş ve CEVd ile enfekteli izolatlar moleküler açıdan da karakterize edilmiştir. Nükleotid dizilimi analizi sonuçları NCBI gen bankasına bildirilen domates, turunçgil ve diğer izolatlara %89-98 sekans benzerliği göstermiştir.

Anahtar Kelimeler:

Viroid, domates, Pospiviroid, RT-PCR, mekanik inokulasyon

Investigation and Molecular Characterization of Viroid Agents Existance at Pospiviroid Genus on Tomato Plant

Study was conducted to investigate viroids’ existance at Pospiviroid genus on tomato plant. Between 2014-2015, 400 tomato seeds, seedlings and leaves samples were collected and total nucleic acid extracts of samples were obtained. In RT-PCR studies which were carried out by Pospi1-FW/RE and Vid-FW/RE primer sets, any viroid infection was detected in 398 tomato samples. Two tomato samples on ‘’Çeri‘’ cultivar, found infected with Citrus exocortis viroid (CEVd) by CEVd-FW/RE primer pair. 371 bp of DNA bands were shown belong to samples in agarose gel electrophoresis studies. CEVd infected isolates were also characterized molecularly. Nucleotid aligments analysis showed %89-98 sequence similiarity to some tomato, citrus and other isolates reported to NCBI Genbank database.

Keywords:

Viroid, tomato, Pospiviroid, RT-PCR, mechanical inoculation,

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