Investigation of Both Drought Stress and In Vitro Culture Conditions Induced Epigenetic Alteration in Genetically Pure Bread Wheat Line Derived from Pehlivan Cultivar

Year 2017, Volume: 45 Issue: 1, 27 - 33, 01.03.2017

Ayşe Şen

Abstract

In the current study; mature bread wheat Triticum aestivum L. embryos from doubled haploid wheat line obtained from Pehlivan cultivar were cultured on drought stress induction medium MS + 0.1 mgL-1 2.4-D + diffe- rent concentrations PEG 6000 0, 10, 20, 40 and 60 gL-1 for 28 days. Effects of drought stress on regenerated plants and methylation alterations due to both drought stress treatment and in vitro culture conditions were investigated in the combination of Inter-Retrotransposon Amplified Polymorphism IRAP with methylation sensitive restriction enzyme MspI and HpaII digestions techniques on 28-day-old control and stress treated regenerated plants. Drought stress decreased the average of plant height and fresh weight of regenerated plants than the controls. A 90% polymorphism rate and 0.6 epigenetic similarity were obtained between cont- rols and drought stress treated experimental groups with the combination of methylation-sensitive restriction enzymes and IRAP. Additionally, the epigenetic similarity ratios between the two controls were detected to 0.733. As a result of these; to investigate methylation alterations due to both in vitro culture conditions and drought stress applications under in vitro conditions can be obtained more realistically performance with using genetically pure line.

Keywords

DNA methylation, drought stress, in vitro screening, doubled haploidy, Wheat

Kuraklık Stresinin ve İn Vitro Kültür Koşullarının Teşvik Ettiği Epigenetik Değişikliğin Pehlivan Çeşidinden Elde Edilen Genetik Olarak Saf Ekmeklik Buğday Hattında Araştırılması

Abstract

B u çalışmada; Pehlivan ekmeklik buğday Triticum aestivum L. çeşidinden elde edilen katlanmış haploid buğday hattının olgun embriyoları, kuraklık stresi teşvik ortamında MS + 0.1 mgL-1 2.4-D + farklı derişimlerde PEG 6000 0, 10, 20, 40 ve 60 gL-1 28 gün boyunca kültüre alınmıştır. Kuraklık stresinin rejenere olan bitkiler üzerine etkileri ve kuraklık stresi uygulaması ve in vitro kültür koşulları nedeniyle oluşan metilasyon değişiklikleri metilasyona duyarlı restriksiyon enzimi MspI ve HpaII kesimi ile Inter-retrotransposon Amplified Polimorfizmi IRAP tekniklerinin kombinasyonu ile 28 günlük kontrol ve stres uygulanan bitkilerde araştırılmıştır. Kuraklık stresi uygulaması rejenere olan embryoların ortalama bitki yüksekliğini ve taze ağırlığını kontrole göre azaltmıştır. Metilasyona duyarlı restriksiyon enzim kesimi ve IRAP tekniklerinin kombinasyonu ile kontrol ve kuraklık stresi uygulanmış deney grupları arasında %90’lık bir polimorfizm oranı ile 0.6’lık bir epigenetik benzerlik elde edilmiştir. Ayrıca, iki kontrol grubu arasındaki epigenetik benzerlik oranı da 0.733 olarak saptanmıştır. Sonuç olarak; hem in vitro kültür ortamının hem de in vitro ortamda kuraklık stresi uygulamasının oluşturduğu metilasyon değişikliklerini incelemek için genetik olarak saf hatların kullanımı ile daha doğru sonuçlar elde edilebilmiştir

Keywords

DNA metilasyonu, kuraklık stresi, in vitro tarama, katlanmış haploidi, buğday

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