Overexpression of jaburetox 2-Ec in potato (Solanum tuberosum L.)
Küçük Resim Yok
Tarih
2022
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Niğde Ömer Halisdemir Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada patates bitkisinde Jaburetox 2-Ec peptidinin ifadesinin artırılarak Tuta absoluta zararlısına karşı dirençli transgenik patates genotiplerinin geliştirilmesi amaçlanmıştır. Bu amaçla, 35S promotoru kontrolündeki hedef peptit dizisini içeren gen kaseti, Agrobakterium-aracılı gen transferi yöntemiyle Lady Olympia çeşidine aktarılmıştır. Deney yürütülürken bitkinin yaprak ve interned kısımları kullanılarak Agrobakterium (EHA 105) ile enfeksiyon sağlanmıştır.Transgenik bitki ve bakteri seçici markörü olarak 50 mg L-1 konsantrasyonlu Neomisin fosfotransferaz (nptII) geni kullanılmıştır.Yapılan moleküler analizler sonucunda hedef diziyi içeren T-DNA bölgesinin bitki genomuna entegre olduğu gösterilmiştir. Ardından yaprak besleme yöntemi ile transgenik bitkilerin ikinci, üçüncü ve dördüncü evre Tuta absoluta larvalarına karşı direnci test edilmiştir. Sonuçta transgenik bitkilerle beslenen larvalarda, larval döneme bağlı olarak 11-33 % arasında ölüm oranı tespit edilmiştir. Ayrıca control bitkilerle beslenen larvalarda ağırlık artışının transgenik bitkilerle beslenen larvalara kıyasla daha yüksek olduğu belirlenmiştir. Bu tez çalışması sonucunda, Jaburetox 2-Ec' nin Tuta absoluta zararlısına karşı insektisidal aktiviteye sahip olduğu, ancak bu etkinin doğrulanması açısından daha fazla araştırmaya ihtiyaç olduğu sonucuna varılmıştır.
In this study, it was aimed to develop transgenic potato genotypes resistant to Tuta absoluta pest by increasing the expression of Jaburetox 2-Ec peptide in potato plant.The gene cassette containing the target peptide sequence under the control of 35S promoter was transferred to potato cultivar Lady Olympia by Agrobacterium-mediated gene transfer method. During the experiment, the leaf and internode parts of the plant were used to infect with Agrobacterium tumefaciens strain (EHA 105). Neomycin phosphotransferase (nptII) gene with 50 mg L-1 concentration was used as a selective marker for transgenic plants and bacteria. As a result of molecular analyzes, it was shown that the T-DNA region containing the target sequence was integrated into the plant genome. Then, the resistance of transgenic potato plants to second, third and fourth instar larvae of Tuta absoluta was tested using leaf feeding bioassay method. As a result, mortality rate of larvae fed on transgenic plants ranged from 11% to 33% depending on the larval stage. In addition, it was determined that the weight gain in larvae fed with control plants was higher than the larvae fed with transgenic plants.As a result of this thesis, it was concluded that Jaburetox 2-Ec has an insecticidal activity against Tuta absoluta pest, but more research is needed to confirm the initial results.
In this study, it was aimed to develop transgenic potato genotypes resistant to Tuta absoluta pest by increasing the expression of Jaburetox 2-Ec peptide in potato plant.The gene cassette containing the target peptide sequence under the control of 35S promoter was transferred to potato cultivar Lady Olympia by Agrobacterium-mediated gene transfer method. During the experiment, the leaf and internode parts of the plant were used to infect with Agrobacterium tumefaciens strain (EHA 105). Neomycin phosphotransferase (nptII) gene with 50 mg L-1 concentration was used as a selective marker for transgenic plants and bacteria. As a result of molecular analyzes, it was shown that the T-DNA region containing the target sequence was integrated into the plant genome. Then, the resistance of transgenic potato plants to second, third and fourth instar larvae of Tuta absoluta was tested using leaf feeding bioassay method. As a result, mortality rate of larvae fed on transgenic plants ranged from 11% to 33% depending on the larval stage. In addition, it was determined that the weight gain in larvae fed with control plants was higher than the larvae fed with transgenic plants.As a result of this thesis, it was concluded that Jaburetox 2-Ec has an insecticidal activity against Tuta absoluta pest, but more research is needed to confirm the initial results.
Açıklama
Fen Bilimleri Enstitüsü, Tarımsal Genetik Mühendisliği Ana Bilim Dalı
Anahtar Kelimeler
Biyoteknoloji, Biotechnology
