‘Superior seedless’ üzüm çeşidinde sürgün ucu büyüklüğünün in vitro rejenerasyon ve virüs eliminasyonu üzerine etkileri

Öz

Bağcılıkta, hücre içi patojenlerin neden olduğu en önemli hastalık etmeni virüslerdir. Bulaşık bitkiler zayıf gelişim, uyanmanın gecikmesi, verim ve kalitede azalma gibi belirtiler gösterebilir hatta birkaç yıl içerisinde ölebilir. Bitkileri virüslerden arındırmak için bazı yöntemler mevcuttur. Termoterapiyle birlikte uygulanan sürgün ucu kültürü en yaygın yöntemdir. Yöntemin başarısı eksplantın büyüklüğüne bağlıdır yani, eksplant ne kadar küçük olursa o kadar fazla virüsten ari bitki elde edilebilir. Öte yandan küçük eksplantların rejenerasyon kapasiteleri düşüktür. Bu çalışmada, ‘Superior Seedless’ üzüm çeşidinde eksplant büyüklüğü ile virüs eliminasyonu arasındaki bağlantı incelenmiştir. Bitkiler termoterapi ve in vitro kültür öncesinde, Double Antibody Sandwich-Enzyme-Linked Immunosorbent Assay (DAS-ELISA) ve Real-time Reverse Transcription-Polymerase Chain Reaction (Real time RT-PCR) teknikleri ile test edilmiş ve farklı virüsler (Grapevine leafroll associated virus -1, -3, Grapevine leafroll associated virus -4 strains, Grapevine rupestris stem pitting associated virus) ile bulaşık oldukları doğrulanmıştır. Farklı büyüklükte (0.1-0.5-1-5-10 mm) sürgün ucu eksplantları, 1 mgl-1 BA içeren MS ortamında kültüre alınmıştır. Küçük boyutlu eksplantlarda virüs bulaşıklığı az olmuş ya da hiç saptanmamıştır.

Anahtar Kelimeler

• in vitro
• eksplant büyüklüğü
• virüs eliminasyonu
• rejenerasyon
• asma

Effects of shoot tip size on in vitro regeneration and virus elimination of grapevine cv. superior seedless

Abstract

In this study, we investigate the relation between the explant size and the success of virus elimination in ‘Superior Seedless’ grape variety. Plants were tested by prior to thermotherapy and in vitro culture by using Double Antibody Sandwich-Enzyme-Linked Immunosorbent Assay (DAS-ELISA) and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and verified as infected by different viruses (Grapevine leafroll associated virus -1+3, Grapevine leafroll associated virus -4 Strains, Grapevine rupestris stem pitting associated virus). Different size of shoot tip explants (0.1-0.5-1-5-10 mm) were excised and cultured on MS medium supplemented with 1 mgl-1 BA. All of the explants bigger than 0.1 mm 100% regenerated while 70% of the 0.1 mm explants showed complete regeneration and turned into a whole plant. Plants regenerated from 10 mm shoot tip explants have all three viruses (GLRaV-1+3, GLRaV-4 strains and GRSPaV) as founded in mother plant. Plants regenerated from 5 mm explants have two of them (GLRaV-1+3, GLRaV-4 strains) and plants regenerated from 1 mm explants have one (GLRaV-1+3). Plants regenerated from 0.1 mm explants were found completely virus-free.

Keywords

• in vitro
• explant size
• virus elimination
• regeneration
• grapevine

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