Developing haploid plants by ectopically expressing homologous BABY BOOM (BBM) genes in sorghum (Sorghum bicolor) and soybean (Glycine max)
Küçük Resim Yok
Tarih
2023
Yazarlar
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Yayıncı
Niğde Ömer Halisdemir Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Haploid bitki geliştirmek için kullanılan konvansiyonel tekniklerin rekalsitrant bitkilerde etkin bir şekilde kullanılamamasından dolayı yeni tekniklerin uygulanması oldukça önem kazanmaktadır. Bunun için özellikle soyada ve sorgumda alternatif yaklaşımlara ihtiyaç duyulmaktaktadır. Yeni sistemlere örnek olarak BABY BOOM (BBM) geninin, çeltik, mısır ve tütünde, darıdaki ortoloğunun ifadelenmesiyle ya da yumurta hücresinde ektopik ifadelenmesiyle haploid indüksiyonu çok yüksek oranlarda teşvik ettiği bilinmektedir. Bu tez çalışmasında, nativ sorgum BBM-like genlerinin ve soya GmBBM1 geninin haploid indüksiyon başarısını gözlemlemek için Arabidopsis'ten klonlanan yumurta hücresine özgü promotör, DD45, kullanılarak yumurta hücresinde ektopik olarak ifadelenmesi sağlanmıştır. BBM genleri ve DD45 promotörü içeren vektörler başarıyla tasarlanmıştır. Ploidi seviyesi akış sitometrisi ile T 0 bitkilerinden elde edilen tohumların çimlendirilmesiyle oluşan T 1 bitkilerinden örnek alınarak ve kök ucundaki kromozom sayısı sayılarak kontrol edilmiştir ve analiz sonucunda, üç farklı transgenik soya hattında, 18-4 (3), 37-4 (Y2) and 46Y2 (3), haploid hücre ve anöploidi ile ilişkili kromozomal anomallikler oluştuğu tespit edilmiştir.
The constraints in application of conventional techniques in order to develop haploid plants in a reproducible manner, it is of high demand implementing the novel approaches in highly recalcitrant crops. An example to existing novel systems is BABY BOOM (BBM) gene for which the expression from pearl millet or its ectopic expression in egg cell in rice, maize and tobacco has known to promote the haploid induction at very high rates. In this thesis work, native sorghum BBM like genes and soybean GmBBM1 gene were ectopically expressed in egg cell in respective crops under egg cell specific promoter, DD45 cloned from Arabidopsis thaliana to observe the haploid induction responses in these crops. The constructs for three BBM inserts under the control of DD45 promoter were succesfully designed. Haploidy level was checked by flow cytometer and chromosome count by root tip squash method on T 1 plants germinated from T 0 seeds in soybean and the analysis showed that chromosomal aberrations related to haploid cells and aneuploidy were appearent in three transgenic soybean lines, 18-4 (3), 37-4 (Y2) and 46Y2 (3).
The constraints in application of conventional techniques in order to develop haploid plants in a reproducible manner, it is of high demand implementing the novel approaches in highly recalcitrant crops. An example to existing novel systems is BABY BOOM (BBM) gene for which the expression from pearl millet or its ectopic expression in egg cell in rice, maize and tobacco has known to promote the haploid induction at very high rates. In this thesis work, native sorghum BBM like genes and soybean GmBBM1 gene were ectopically expressed in egg cell in respective crops under egg cell specific promoter, DD45 cloned from Arabidopsis thaliana to observe the haploid induction responses in these crops. The constructs for three BBM inserts under the control of DD45 promoter were succesfully designed. Haploidy level was checked by flow cytometer and chromosome count by root tip squash method on T 1 plants germinated from T 0 seeds in soybean and the analysis showed that chromosomal aberrations related to haploid cells and aneuploidy were appearent in three transgenic soybean lines, 18-4 (3), 37-4 (Y2) and 46Y2 (3).
Açıklama
Fen Bilimleri Enstitüsü, Tarımsal Genetik Mühendisliği Ana Bilim Dalı
Anahtar Kelimeler
Ziraat, Agriculture
