Korungaya (Onobrychis viciifolia Scop.) Partikül Bombardımanı İle Gen Aktarımında Fiziksel ve Kimyasal Parametrelerin Etkisi

Bu çalışma partikül bombardımanı sistemi kullanılarak korunga bitkisine (Onobrychis viciifolia Scop) gen aktarımında uygun parametrelerin belirlenmesi amacıyla Ankara Üniversitesi, Ziraat Fakültesi, Tarla Bitkileri Bölümü Biyoteknoloji laboratuarında yürütülmüştür. Çalışmada, eksplant olarak kotiledonlar, mikrotaşıyıcı olarak 1.0 µm çapındaki altın parçacıkları ve belirleyici (markör) gen olarak ise “pB1221.23”plazmidinde bulunan ßglucuronidase (GUS) geni kullanılmıştır. Bombardıman, üç ayrı yöntemle hazırlanan altın partikülleri ile, farklı iki fırlatılma basıncı (1100 ve 1350 psi) ve 6 cm uzaklıktan Bio-Rad®/PDS-1000/He gen transformasyon sistemi kullanılarak gerçekleştirilmiştir. Elde edilen veriler değerlendirildiğinde, korunga kotiledonlarına partikül bombardımanı ile gen aktarımında, doğrudan gliserol ile hazırlanan mikrotaşıyıcıların 1350 psi’lik basınçta fırlatılması ile en iyi sonucu verdiği saptanmıştır

Anahtar Kelimeler:

Korunga, Partikül Bombardımanı, Gen Aktarımı, GUS

Effect of Physical and Chemical Parameters on Transient Gene Expression via Particle Bombardment in Sainfoin (Onobrychis viciifolia Scop.)

This study was carried out at the biotechnology laboratories of Department of Field Crops, Faculty of Agriculture, Ankara University to determine suitable transformation parameters and investigate marker gene transfer possibilities to sainfoin (Onobrychis viciifolia Scop.) using the particle bombardment technique. Sainfoin plant cotyledons were used as plant material; 1.0 µm diameter gold particles as microprojectiles; the plasmid “pB1221.23” containing the ß-glucuronidase (GUS) as marker gene were used in this study. Microprojectiles were prepared in three different methods (directly glycerol, alcohol + water, 3 alcohol + water) Bio-Rad®/PDS-1000 /He gene transfer system was used for bombardment in 1100 and 1350 psi pressures and 6 cm distance. The statistical analysis of results revealed that microprojectiles directly prepared with glycerol and 1350 psi pressure were suitable for particle bombardment delivery marker gene transfer to sainfoin cotyledons.

Keywords:

Onobrychis viciifolia Scop., Particle Bombardment, Transformation, GUS,

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