Publication: Melez kabak (Cucurbita pepo L.) genotiplerinde in vitro rejenerasyon
Abstract
Melez Kabak (Cucurbita pepo L.) Genotiplerinde in vitro RejenerasyonCucurbita pepo L. Magnoliopsida sınıfı, Cucurbitales takımı, Cucurbitaceae familyasıiçerisinde yer alan, otsu yapıda gövdeye, geniş yapraklara ve yüzeysel dağılan kökleresahip tek yıllık bir bitkidir. Cucurbita pepo L. yabancı tozlanan bir bitki olduğu içingeleneksel ıslah yöntemleri ile seleksiyon çalışmaları uzun sürmektedir. Türkiye’deıslah çalışmaları geleneksel yöntemlerle sürdürülmektedir. Kabakta geliştirilen dokukültürü tekniklerinin kullanılması çeşit ıslahı çalışmalarını hızlandırmasının yanı sırayüksek verim, kalite ve erkencilik yönünden de önemli avantajlar sağlamaktadır. Bu tezçalışması ile Trakya Tarımsal Araştırma Enstitüsünce zararlılardan korunması veüretiminin arttırılması amaçlanan beş farklı hibrit kabak çeşidine ait (Ardendo, Angelina,Torpido, Roni, Sena Hanım) tohumlar kullanılarak in vitro elde edilen fidelerdengelişen kotiledon nodu, nod, sürgün ucu, hipokotil ve internod eksplantları kullanılarakkabak bitkisinin in vitro rejenerasyonu elde edilmiştir. Tüm genotiplerde doku kültürüuygulamaları öncesi ve sonrası kromozom analizleri ile ploidi seviyeleri 2n=40 olarakdoğrulanmıştır. En iyi gövde rejenerasyonu %92,98 ile Ardendo genotipinin kotiledonnodunda sağlanmış ve rejenerasyon için en verimli eksplant kotiledon nodu olmuştur.En iyi gövde rejenerasyonu yanıtı veren besi ortamı ise 1 mg/ L Benzil amino purin(BAP) içeren Murashige ve Skoog (MS) besi ortamı olmuştur. Tüm genotiplerdekotiledon nodları ve sürgün uçları bitki büyüme düzenleyicisi içermeyen MS ve 1 mg/ LIndol Bütirik Asit (IBA) içeren MS besi ortamında başarıyla köklendirilmiştir. Bunlaraek olarak anter ve ovül kültürü ile haploid bitki eldesi için kallus indüksiyonugerçekleştirilmiştir. Anter kültüründe en yüksek kallus rejenerasyonu vereneksplant %88,24 ile Torpido genotipinde 2 mg/ L 2,4-diklorofenoksiasetik asit (2,4-D)içeren besi ortamında sağlanmıştır. Ovül kültüründe ise en yüksek kallusrejenerasyonu %80,77 ile Angelina genotipinde ve 5 mg/ L 2,4-D içeren besi ortamındasağlanmıştır. Elde edilen bulgular ile genotiplerde mikroçoğaltım için en uygun eksplanttipi ve besi ortamı belirlenmiş anter ve ovüllerdan elde edilen kalluslar ile haploid bitkieldesi için önemli bir adım atılmıştır.
In vitro Regeneration in Hybrid Cucurbita pepo L. GermplazmCucurbita pepo L. is an annual plant and member of Magnoliopsida class, Cucurbitalesorder, Cucurbitaceae family. Cucurbita pepo L. has herbaceous body structure, wideleaves and superficial roots. With conventional methods breeding studies last longbecause cucurbits are outbreeding species. In Turkey, breeding studies were conductedwith conventional methods. Using plant tissue culture methods in cucurbit breedingstudies provides important advantages like high yield, quality and rapid production ofnew varieties. In this thesis project, five different hybrid cucurbit variety (Ardendo,Angelina, Torpido, Roni and Sena Hanim) were used to promote national cucurbitabreeding studies which conducted by Thrace Agricultural Research Institute (TARI).Cotyledon, nod, shoot apex and hypocotyl explants which obtained from in vitroproduced sterile plantlets were used as plant material for regeneration studies.Chromosome analyzes were made for all the genotypes and polyploidy levels wereverified (2n=40) before starting the tissue culture studies. The highest shootregenerations were obtained from cotyledon nodes of Ardendo variety with 92,98% andit’s found that the highest regeneration yield was obtained from cotyledon nodes. Thehighest shoot regeneration response was obtained from the Murashige and Skoog (MS)medium with the addition of 1m g/ L benzyl aminopurine (BAP). For all genotypes,cotyledon nodes and shoot apexes were successfully rooted in MS and MS + 1 mg/ LIBA (indole 3 butyric acid). Also, valuable pre-findings were obtained for haploidproduction with anther and ovary culture in cucurbita species. Also, pre-studies weremade for haploid production with anther and ovary culture and callus inductions wereprovided. In anther culture studies, Torpedo has produced the highest callusregeneration yield with 88,24% in the MS + 2 mg/ L 2,4-D (2,4-dichlorofenoxyaceticacid). In ovary culture studies, Angelina genotype has produced the highest callusregeneration yield with 80,77% in the MS + 5 mg/ L 2,4-D. With obtained findings theoptimal explant type and culture medium were determined for micropropagation andcalli which obtained from anther and ovary culture become the valuable starting pointfor haploid plant production.
In vitro Regeneration in Hybrid Cucurbita pepo L. GermplazmCucurbita pepo L. is an annual plant and member of Magnoliopsida class, Cucurbitalesorder, Cucurbitaceae family. Cucurbita pepo L. has herbaceous body structure, wideleaves and superficial roots. With conventional methods breeding studies last longbecause cucurbits are outbreeding species. In Turkey, breeding studies were conductedwith conventional methods. Using plant tissue culture methods in cucurbit breedingstudies provides important advantages like high yield, quality and rapid production ofnew varieties. In this thesis project, five different hybrid cucurbit variety (Ardendo,Angelina, Torpido, Roni and Sena Hanim) were used to promote national cucurbitabreeding studies which conducted by Thrace Agricultural Research Institute (TARI).Cotyledon, nod, shoot apex and hypocotyl explants which obtained from in vitroproduced sterile plantlets were used as plant material for regeneration studies.Chromosome analyzes were made for all the genotypes and polyploidy levels wereverified (2n=40) before starting the tissue culture studies. The highest shootregenerations were obtained from cotyledon nodes of Ardendo variety with 92,98% andit’s found that the highest regeneration yield was obtained from cotyledon nodes. Thehighest shoot regeneration response was obtained from the Murashige and Skoog (MS)medium with the addition of 1m g/ L benzyl aminopurine (BAP). For all genotypes,cotyledon nodes and shoot apexes were successfully rooted in MS and MS + 1 mg/ LIBA (indole 3 butyric acid). Also, valuable pre-findings were obtained for haploidproduction with anther and ovary culture in cucurbita species. Also, pre-studies weremade for haploid production with anther and ovary culture and callus inductions wereprovided. In anther culture studies, Torpedo has produced the highest callusregeneration yield with 88,24% in the MS + 2 mg/ L 2,4-D (2,4-dichlorofenoxyaceticacid). In ovary culture studies, Angelina genotype has produced the highest callusregeneration yield with 80,77% in the MS + 5 mg/ L 2,4-D. With obtained findings theoptimal explant type and culture medium were determined for micropropagation andcalli which obtained from anther and ovary culture become the valuable starting pointfor haploid plant production.