Protoplast Kültürü ve Füzyonu

Yazarlar

Mehmet Örgeç; Ömer Can Ünüvar; Özge Kaya; Songül Gürel; Ekrem Gürel

Özet

Protoplast kültürü ve füzyonu bitki biyoteknolojisi çalışmalarında kullanılan önemli tekniklerden biridir. Hücre duvarı enzimatik ve/veya mekanik yöntemler ile uzaklaştırılmış bitki hücreleri protoplast olarak tanımlanır. Protoplastlar uygun kültür koşullarında yeni hücre duvarı sentezleyebilmekte, mitotik bölünmeye girebilmekte ve tam bitki rejenerasyonu sağlayan kallus dokularına dönüşebilmektedirler. Bu bölümde protoplast çalışmalarının tarihçesi, protoplast izolasyonu, saflaştırılması, kültürü, rejenerasyonu ve füzyonu ile ilintili prensipler ayrıntılı bir şekilde ele alınmıştır. Eksplant tipi ve yaşı, kullanılan enzim kombinasyonları, ozmotik dengeleyiciler, inkübasyon süresi ve fiziksel uygulamalar protoplast izolasyonunu etkileyen faktörlerdir.  Bu parametrelerin optimizasyonunun protoplast verimi, canlılığı ve rejenerasyon kapasitesi üzerindeki kritik etkileri vurgulanmıştır. Ayrıca saflaştırma aşamalarında tekniklerin avantaj ve dezavantajları karşılaştırmalı olarak değerlendirilmiştir. Protoplast rejenerasyonunun büyük ölçüde genotipe bağımlı olduğu, bu nedenle türler arasında evrensel bir protokol geliştirmenin güçlüğü ortaya konmuştur. Protoplast füzyon teknikleri ele alınarak PEG aracılı kimyasal füzyon, elektrofüzyon ve CRISPR/Cas9 tabanlı genom düzenleme yaklaşımlarının bitki ıslahı, somatik hibridizasyon ve fonksiyonel genomik çalışmalardaki önemi tartışılmıştır.  Şeker pancarı, buğday, pirinç, Brassica türleri ve turunçgiller üzerinde gerçekleştirilen uygulama örnekleri protoplast teknolojisinin sunduğu stratejik potansiyeli açıkça ortaya koymaktadır. Genel olarak protoplast kültürü ve füzyonu geleneksel ıslah yöntemlerinin sınırlamalarını aşan ve bitki biyoteknolojisine etkili bir şekilde entegre edilebilen vazgeçilmez araçlar olarak ortaya çıkmaktadır.

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