TAILIEUCHUNG - Transient eexpression of β-glucuronidase reporter gene in sainfoin (Onobrychis viciifolia Scop.) cotyledons via microprojectile bombardment

Transient β-glucuronidase reporter gene (GUS) expression was visualised in whole cotyledons of sainfoin (Onobrychis viciifolia Scop.) after microprojectile bombardment with DNA. | Turk J Biol 25 (2001) 171-176 © TÜB‹TAK Transient Eexpression of β-glucuronidase Reporter Gene in Sainfoin (Onobrychis viciifolia Scop.) Cotyledons Via Microprojectile Bombardment Sertaç ÖNDE Department of Biological Sciences, Faculty of Arts and Sciences, Middle East Technical University, 06531 Ankara - TURKEY Cengiz SANCAK, Suzan ALTINOK, Melahat B‹RS‹N, Murat ÖZGEN Department of Field Crops, Faculty of Agriculture, Ankara University, 06110 D›flkap›, Ankara - TURKEY Received: Abstract: Transient β-glucuronidase reporter gene (GUS) expression was visualised in whole cotyledons of sainfoin (Onobrychis viciifolia Scop.) after microprojectile bombardment with DNA. Optimum rupture disk pressure and sample plate distance were determined to be 1350 psi and 6 cm, respectively. Mean gold particle diameter of µm and 48 h post-bombardment histochemical GUS assay was found to be superior to µm gold particles and 120 h postbombardment GUS assay. Microscopic analysis revealed the localisation of the gold particles predominantly in the epidermal cell layer, whereas less than 1% of the particles were localised in subepidermal cells, as deep as three cell layers. Availability of a regeneration protocol from sainfoin cotyledons and the potential of the microprojectile bombardment recommend future applications for obtaining transgenic sainfoin plants. Key Words: Onobrychis viciifolia Scop., sainfoin, transient gene expression, biolistic, particle bombardment Mikroprojektil Bombard›man› ile Korunga (Onobrychis viciifolia Scop.) Bitkisinde Geçici β-glukuronidaz Reportör gen ‹fadesi Özet: Tüm korunga (Onobrychis viciifolia Scop.) kotiledonlar›na plazmid DNA’s› ile gerçeklefltirilen mikroprojektil bombard›man› sonras›nda geçici β-glukuronidaz (GUS) reportör gen ifadesi gözlemlendi. Optimum bas›nç ve örnek-tabla mesafesi 1350 psi ve 6 cm olarak belirlendi. Ortalama alt›n partikül çap› olarak µm ve bombard›mandan 48 saat sonra yap›lan .

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