Ters lale (Fritillaria imperialis L.) bitkisinde ın vitro koşullarda organ rejenerasyonlarının sağlanması
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Dosyalar
Tarih
2015
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi, Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Liliaceae familyasından Fritillaria cinsine bağlı olan ters lale (Fritillaria imperialis L.), ülkemizde yarı endemik olarak yetişen bir türdür. F. imperialis, çok uzun zamandır bahçe süs bitkisi olarak kullanıldığından başta Hollanda olmak üzere Avrupa'ya ihraç edilmektedir ve doğadan sökümü yasaktır. Bu tez çalışmasında, vejetatif ve generatif çoğaltım işlemlerinde pek çok kısıt barındıran F. imperialis türünün, çeşitli in vitro besin ortamlarında ve farklı kültür koşullarında doku kültürü teknikleri ile çoğaltımı hedeflenmiştir. Bu amaçla, F. imperialis L. bitkisinden alınan çeşitli eksplantlardan (soğan, yaprak, gövde, kök), in vitro koşullarda organ rejenerasyonlarının gerçekleştirilmesine çalışılmıştır. Sürgünler üzerindeki yeşil yaprak ekplantları için en yüksek kallus gelişim oranı (%59.26) 1.0 mg/l BAP + 0.5 mg/l NAA + 0.4 mg/l IAA ile 60 g/l sükroz içeren B5 besin ortamında, petal yaprak eksplantlarında ise (%33.33) aynı içeriğe sahip MS besin ortamında tespit edilmiştir. Yeşil yaprak eksplantları ile yapılan çalışmalarda; MS temelli besin ortamlarında rejenere olan kallusların, B5 temelli besin ortamında rejenere olanlara göre hem somatik embriyo (sırasıyla en yüksek embriyogenik kallus oranları %92.31 ve %56.25), hem de sürgün rejenerasyonuna yönelme bakımından (sırasıyla en yüksek sürgün oluşturan kallus oranları %88.89 ve 37.50) daha uygun oldukları kaydedilmiştir. Bu ortamlar arasında ise oksinlerin 0.5 mg/l NAA + 0.4 mg/l IAA), yüksek dozda BAP (1.0 mg/l) ile kombine edildikleri MS ve B5 besin ortamlarında sürgün oluşturan kallus oranları (sırasıyla %88.89 ve %37.50), daha düşük BAP (0.1 mg/l) konsantrasyonunda gelişenlere (sırasıyla %53.85 ve %16.67) göre daha başarılı bulunmuşlardır. Başlangıç materyali olarak yeşil yaprak eksplantlarının kullanıldığı çalışmalarda, dolaylı organogenez bakımından oksinlerle birlikte besin ortamına eklenen BAP sitokinini sürgün rejenerasyonunda, Kin veya TDZ sitokininleri ise genellikle kallus uyarımında daha etkili bulunmuşlardır. F. imperialis türünde, depolama süresince soğanlar üzerinde kendiliğinden gelişen köklerin ilk defa başlangıç materyali olarak kullanıldığı ve 1.0 mg/l NAA + 2.0 mg/l 2,4-D içeren MS besin ortamının ele alındığı denemelerde ise kallus rejenerasyonu bakımından 16 saat aydınlık/8 saat karanlık fotoperiyot koşullarının (%33.33), karanlık koşullara göre daha elverişli olduğu ortaya konulmuştur. Yeşil yaprak eksplantları altkültüre alındıktan 6 hafta sonra toplamda 146 adet olmak üzere yeşil yaprak eksplantı başına 1.35 adet sürgün rejenere olmuştur. Kültürde 36 hafta boyunca altkültüre alınmadan inkübe edilen yeşil yaprak eksplantlarından gelişen kalluslar üzerinde toplamda 62 adet soğancık rejenerasyonu gerçekleşirken, petal yapraklarının eksplant kaynağı olduğu denemeler sonucunda ise soğancık gelişimi gözlenmemiştir. 36 haftalık kültür periyodu sonunda yeşil yaprak eksplantı başına 1.89 adet soğancık (FS7: B5 + 1.0 mg/l BAP + 0.5 mg/l NAA + 0.4 mg/l IAA besin ortamında) elde edilmiştir.
Crown imperial (Fritillaria imperialis L.) which belongs to Fritillaria genus of Liliaceae family, is grown as semi-endemic in our country. Since crown imperial (F. imperialis L.) is used as ornamental plant for a long time because of its splendid flowers, it is exported to Europe especially Holland and it is forbidden to remove from nature. Aim of this study is to propagate using various in vitro nutrient mediums and different tissue culture techniques in F. imperialis species, which includes lots of restrictions in both vegetative and generative production. For this purpose, organ regeneration under in vitro conditions was performed by using various explants (bulb, leave, shoot, root) of F. imperialis L. For the green leaf explants, the maximum callus formation ratio (59.26%) was obtained on B5 medium containing 1.0 mg/l BAP + 0.5 mg/l NAA + 0.4 mg/l IAA and 60 g/l sucrose, while for the petal explants, the maximum callus formation ratio (33.3%) was detected on MS medium with the same content. In the studies was made by using the green leaf explants, it is recorded that regenerated callus in MS based nutrient media had more tendency to form both somatic embryo (the maximum embryogenic callus ratios 92.31% and 56.25%, respectively) and shoot regeneration (the maximum shoot forming callus ratios 88.89% and 37.50%, respectively) than regenerated callus in B5 based nutrient media. Among these media, MS and B5 media supplemented with higher concentration of BAP (1.0 mg/l) combined with auxins (0.5 mg/l NAA + 0.4 mg/l IAA) were more successful (88.89% and 37.50%, respectively) than low concentration BAP (0.1 mg/l) combined with same auxins (53.85% and 16.67%, respectively) regarding callus ratios forming shoot. In the studies which used green leaf explants as starting material, the medium supplemented with BAP cytokinin and auxins was more efficient in shoot regeneration, while the medium containing Kin or TDZ cytokinins and auxins was more effective in callus stimulation. At the F. imperialis species that using MS medium include 1 mg/l NAA + 2 mg/l 2,4-D and roots as initial explants the first time which was grown by themselves on bulbs during storage and, photoperiod conditions of 16 hours light/8 hours dark were more suitable than fully dark conditions in terms of callus regeneration. The green leaf explants were subculture after six weeks of culture, 1.35 shoots per green leaf explant and total 146 shoots regenerated. Total 62 bulblets were obtained from callus that developed on green leaf explants which maintained 36 weeks in culture without subculture, while the petal leaves formed no bulblets. At the end of the 36 weeks culture periods, 1.89 bulblets were obtained per green leaf explant (at the FS7 coded medium which was included B5 + 1 mg/l BAP + 0.5 mg/l NAA + 0.4 mg/l IAA).
Crown imperial (Fritillaria imperialis L.) which belongs to Fritillaria genus of Liliaceae family, is grown as semi-endemic in our country. Since crown imperial (F. imperialis L.) is used as ornamental plant for a long time because of its splendid flowers, it is exported to Europe especially Holland and it is forbidden to remove from nature. Aim of this study is to propagate using various in vitro nutrient mediums and different tissue culture techniques in F. imperialis species, which includes lots of restrictions in both vegetative and generative production. For this purpose, organ regeneration under in vitro conditions was performed by using various explants (bulb, leave, shoot, root) of F. imperialis L. For the green leaf explants, the maximum callus formation ratio (59.26%) was obtained on B5 medium containing 1.0 mg/l BAP + 0.5 mg/l NAA + 0.4 mg/l IAA and 60 g/l sucrose, while for the petal explants, the maximum callus formation ratio (33.3%) was detected on MS medium with the same content. In the studies was made by using the green leaf explants, it is recorded that regenerated callus in MS based nutrient media had more tendency to form both somatic embryo (the maximum embryogenic callus ratios 92.31% and 56.25%, respectively) and shoot regeneration (the maximum shoot forming callus ratios 88.89% and 37.50%, respectively) than regenerated callus in B5 based nutrient media. Among these media, MS and B5 media supplemented with higher concentration of BAP (1.0 mg/l) combined with auxins (0.5 mg/l NAA + 0.4 mg/l IAA) were more successful (88.89% and 37.50%, respectively) than low concentration BAP (0.1 mg/l) combined with same auxins (53.85% and 16.67%, respectively) regarding callus ratios forming shoot. In the studies which used green leaf explants as starting material, the medium supplemented with BAP cytokinin and auxins was more efficient in shoot regeneration, while the medium containing Kin or TDZ cytokinins and auxins was more effective in callus stimulation. At the F. imperialis species that using MS medium include 1 mg/l NAA + 2 mg/l 2,4-D and roots as initial explants the first time which was grown by themselves on bulbs during storage and, photoperiod conditions of 16 hours light/8 hours dark were more suitable than fully dark conditions in terms of callus regeneration. The green leaf explants were subculture after six weeks of culture, 1.35 shoots per green leaf explant and total 146 shoots regenerated. Total 62 bulblets were obtained from callus that developed on green leaf explants which maintained 36 weeks in culture without subculture, while the petal leaves formed no bulblets. At the end of the 36 weeks culture periods, 1.89 bulblets were obtained per green leaf explant (at the FS7 coded medium which was included B5 + 1 mg/l BAP + 0.5 mg/l NAA + 0.4 mg/l IAA).
Açıklama
Anahtar Kelimeler
Ters Lale, Fritillaria imperialis, İn vitro,, Soğancık, Rejenerasyon
