Hatice ÇÖLGEÇEN, Ufuk KOCA, Gülnur TOKER

Infl uence of diff erent sterilization methods on callus initiation and production of pigmented callus in Arnebia densifl ora Ledeb

Bu çalışmada sodyum hipoklorit ve Bitki Koruyucu Karışımın yüzey sterilizasyonundaki etkileri araştırılmıştır. Antibiyotik-antimikotik çözeltisinin kültür ortamına eklenerek kallus indüksiyonundaki etkilerini inceledik. Eksplantlar başlangıç olarak farklı sodyum hipoklorit konsantrasyonlarıyla steril edilerek kinetin (0,29 μM) ve naft alen asetik asit (NAA,10 μM) içeren MS ortamında kültüre alınmıştır. Eksplantlardaki kontaminasyonlar veya yüksek sodyum hipoklorit konsantrasyonundan dolayı kallus oluşumu gözlenmemiştir. Farklı konsantrasyonlardaki Bitki Koruyucu Karışımı ve antibiyotikler kontaminasyonu azaltmış ve sürgün ucu ve genç kök eksplantlarında kallus indüksiyonu gerçekleşmiştir. En iyi kallus oluşumu % 1-2 Bitki Koruyucu Karışım kullanımında bulunurken, sürgün uçlarındaki kallus indüksiyonu yüksek konsantrasyonda (% 4) azalmıştır. Bu çalışma, doğal ortamında arazide yetişen A. densifl ora’nın eksplant kontaminasyonlarının azaltılması ve başarılı sterilizasyon ile ilgili ilk çalışmadır. Oluşturulan kallus kültürleri renkli kalluslar üretti bunlar da spektrofotometrik yöntemle analiz edildi.

Arnebia densifl ora Ledeb.’de kallus oluşumunda farklı sterilizasyon metodlarının etkisi ve renkli kallus üretimi

We analyzed the eff ects of sodium hypochlorite and Plant Preservative Mixture (PPM) on surface sterilization. We also examined the eff ects that the addition of an antibiotic-antimycotic solution to the culture medium had on callus induction. Explants were initially sterilized with diff erent concentrations of sodium hypochlorite and cultured on MS media containing kinetin (0.29 μM) and naphthalene acetic acid (NAA, 10 μM). No calluses were produced, either because of contamination of the explants, or loss of explants as a result of the high levels of sodium hypochlorite. Th e application of PPM and antibiotics at diff erent concentrations reduced contamination and led to callus induction from shoot apexes and young root explants. Th e best callus responses were obtained using PPM at 1%-2%, whereas callus induction on shoot apexes diminished at higher concentrations (4% PPM). Th is is the fi rst report of successful sterilization and reduced contamination of explants from naturally fi eld grown A. densifl ora by PPM. Moreover, established callus cultures produced pigmented calluses, which were analyzed spectrophotometrically.

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