Balcı aspir (Carthamus tinctorius L.) çeşidinin mikroçoğaltımı
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2018-04-27
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Batman Üniversitesi Fen Bilimleri Enstitüsü
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info:eu-repo/semantics/openAccess
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Attribution-ShareAlike 3.0 United States
Özet
Aspir (Carthamus tinctorius L.), gerek beslenme gerekse de biyoyakıt üretiminde öne çıkan yağlı tohumlu bitkilerin başında gelmektedir. Bu tez çalışmasında, 2011 yılında tescil edilen ve linoleik tipte olan Balcı aspir çeşidinin olgun tohumlarından itibaren optimum mikroçoğaltım prosedürünün oluşturulması amaçlandı. Aspir bitkisinin in vitro yetiştirme koşullarını belirlemek amacıyla, ilk olarak tohumların sterilizasyonunun sağlanması için sodyum hipokloritte (NaOCI) bekletme süreleri test edildi. %5'lik NaOCI'in farklı sürelerinde (10, 15, 20, 25, 30, 40,50, 60 dk) ayrı ayrı bekletilen tohumlar çatlatılan ve çatlatılmayan olmak üzere 2 farklı şekilde kültüre alındı. Enfeksiyon riskinin azaltılması için tohumların %5'lik NaOCl'de minimum 60 dakika bekletilmesi gerektiği belirlendi. Çatlatılmış tohumların tamamının çimlendiği, çatlatılmamış tohumların ise çok az sayıda (%16.6) çimlendiği görüldü. İn vitro çimlenme için tohumların mutlaka çatlatıldıktan sonra kültüre bırakılması gerektiği tespit edildi. Aspir tohumlarının in vitro çimlendirilmesine, MS gücünün (kontrol, 1/1, ½, ve ¼), karbon kaynağının (30 g/L sakkaroz, maltoz ve fruktoz) ve ışığın (karanlık ve aydınlık -16/8 fotoperiyod) etkisi ayrı ayrı incelendi. MS besi ortamındaki tohumların kontrol grubuna (MS içermeyen) göre daha iyi geliştiği ve morfolojik gelişimde gözönünde bulundurulduğunda ¼ MS'in en uygun besi ortamı olduğu belirlendi. Bununla birlikte besi ortamında kullanılan şeker çeşidinin etkili olduğu ve sakkarozun diğer şekerlerden daha iyi sonuç verdiği saptandı. Aspirin hem aydınlık hem de karanlık ortamda çimlendiği ancak çimlenen tohumların aydınlık ortamda daha iyi gelişme gösterdiği belirlendi. Tüm bu sonuçlar değerlendirildiğinde, tohumlar için en iyi çimlenme koşulları; 30 g sakkaroz ile desteklenmiş ¼ MS besi ortamında ve aydınlıkta (16/8 fotoperiyod) kültüre alınan çatlatılmış tohumlarda olduğu tespit edildi. Mikroçoğaltma çalışmalarında, in vitro koşullarda çimlendirilen tohumlardan elde edilen steril fideler kullanıldı. Sürgün çoğaltılmasında BAP ve Kinetinin (Kin) farklı konsantrasyonlarının (0.5, 1.0, 2.0, 4.0 mg/L) etkisi araştırıldı. Sitokinin içermeyen kontrol grubu dışında test edilen BAP ve Kin oranlarının tümünde yeni sürgün oluşumu gözlendi. BAP konsantrasyonları arasında en iyi sürgün çoğaltımı, eksplant başına yaklaşık 5.33 adet sürgün ile 0.5 mg/L BAP'lı ortamdan elde edilirken, Kin uygulamalarında sırasıyla 3.75 - 3.33 adet sürgün ile 0.5 ve 4.0 mg/L Kin içeren ortamdan elde edildi. Ancak BAP uygulamalarının genelinde ve 4.0 mg/L Kin uygulamasında oluşan sürgünlerde vitrifikasyon olduğu belirlendi. Bu nedenle, vitrifikasyonun olmadığı köklenmeye uygun sağlıklı sürgünlerin geliştiği 0.5 mg/L Kin ile desteklenmiş MS besi ortamının balcı aspir çeşidinin in vitro sürgün çoğaltımı için en ideal ortam olduğu tespit edildi. İn vitro sürgünlerin köklendirilmesi amacıyla MS besi ortamına NAA (0.5, 1.0, 2.0 mg/L) ve IAA (0.5, 1.0, 2.0 mg/L) in farklı konsantrasyonları ilave edildi. 0.5 mg/L NAA ve IAA hariç tüm gruplarda sürgünlerin köklendiği ve en iyi kök oluşumunun, eksplant başına 20 adet ile 2.0 mg/L NAA içeren ortamda kültüre alınan sürgünlerde olduğu tespit edildi. Elde edilen köklü fideler torf – perlit karışımı içeren saksılara dikilerek toprağa adaptasyonu sağlandı. Sonuç olarak bu tez çalışmasında, Balcı aspir (Carthamus tinctorius L.) çeşidinin olgun tohumlarından itibaren in vitro çimlenme, sürgün çoğaltımı ve köklendirilme için ideal bir protokol oluşturuldu.
Safflower (Carthamus tinctorius L.) is at the begining of oilseed crops that are important both in biofuel production and nutritionally. In this thesis study, it was aimed to establish an optimum microproduction process from the mature seeds of Balci safflower variety which is linoleic type and was registered in 2011. In order to determine the optimum conditions for in vitro growth, the safflower seeds were separately kept at different times (10, 15, 20, 25, 30, 40, 50, 60 min) in 5% sodium hypochlorite (NaOCI) and were cultured in two different applications with their seeds broken or unbroken. It was observed that all of broken seeds were germinated and only a small part of the unbroken seeds (16.6%) were germinated. For this reason, it was determined that seeds should definitely be cultured for in vitro germination after brokening. It was also determined that the seeds should be kept at 5% NaOCl for a minimum of 60 minutes to reduce the risk of infection. The effects of MS strength (control, 1/1, ½, and ¼), carbon source (30 g/L sucrose, maltose and fructose) and light (dark and light-16/8 photoperiod) on in vitro germination of safflower seeds were separately examined. It was determined that seeds in the MS medium were better developed than the control group (without MS) and ¼ MS is the optimal medium for the morphological development of the seeds. However, it was found that sugar content used in the medium was effective and sucrose was better than other sugars. It has been determined that safflower is germinated in both light and dark conditions, but germinated seeds are better developed in the light conditions. When all these results were evaluated, it was determined that the best germination rate of seeds was in ¼ MS medium supplemented with 30 g sucrose and in broken seeds cultured in light (16/8 photoperiod). In the micropropagation studies, sterile seedling were used as a material which obtained from seeds germinated in in vitro conditions. The effects of different concentrations of BAP and Kinetin (0.5, 1.0, 2.0, 4.0 mg/L) on shoot multiplication were investigated. New shoot formation was observed in all of the BAP and kinetin concentrations tested except the control group (without cytokine). The best shoot multiplication was obtained from medium with 0.5 mg/L BAP with approximately 5.33 shoots per explant among BAP applications. On the other hand 3.75 - 3.33 shoots per explant were obtained from medium containing 0.5 and 4.0 mg/L Kin in kinetin applications, However, vitrification was observed in shoots at all of the BAP and 4.0 mg/L Kinetin applications. Therefore, MS medium supplemented with 0.5 mg/L Kin was determined the optimum medium (rooting-suitable, healthy shoots, without vitrification) for in vitro shoot propagation of Balcı safflower. Different concentrations of NAA (0.5, 1.0, 2.0 mg/L) and IAA (0.5, 1.0, 2.0 mg/L) were added to the MS medium for rooting in vitro shoots. Generally all shoots were rooted in all applications except 0.5 mg/L NAA and IAA. The best root formation was observed in medium containing 2.0 mg/L NAA with 20 roots to per explant. The soil adaptation of the obtained rooted seedlings was provided with planted in pots containing a mixture of peat and perlite. As a result in this thesis study, an optimum protocol was established for in vitro germination, shoot propagation and rooting from mature seeds of Balcı safflower (Carthamus tinctorius L.) variety.
Safflower (Carthamus tinctorius L.) is at the begining of oilseed crops that are important both in biofuel production and nutritionally. In this thesis study, it was aimed to establish an optimum microproduction process from the mature seeds of Balci safflower variety which is linoleic type and was registered in 2011. In order to determine the optimum conditions for in vitro growth, the safflower seeds were separately kept at different times (10, 15, 20, 25, 30, 40, 50, 60 min) in 5% sodium hypochlorite (NaOCI) and were cultured in two different applications with their seeds broken or unbroken. It was observed that all of broken seeds were germinated and only a small part of the unbroken seeds (16.6%) were germinated. For this reason, it was determined that seeds should definitely be cultured for in vitro germination after brokening. It was also determined that the seeds should be kept at 5% NaOCl for a minimum of 60 minutes to reduce the risk of infection. The effects of MS strength (control, 1/1, ½, and ¼), carbon source (30 g/L sucrose, maltose and fructose) and light (dark and light-16/8 photoperiod) on in vitro germination of safflower seeds were separately examined. It was determined that seeds in the MS medium were better developed than the control group (without MS) and ¼ MS is the optimal medium for the morphological development of the seeds. However, it was found that sugar content used in the medium was effective and sucrose was better than other sugars. It has been determined that safflower is germinated in both light and dark conditions, but germinated seeds are better developed in the light conditions. When all these results were evaluated, it was determined that the best germination rate of seeds was in ¼ MS medium supplemented with 30 g sucrose and in broken seeds cultured in light (16/8 photoperiod). In the micropropagation studies, sterile seedling were used as a material which obtained from seeds germinated in in vitro conditions. The effects of different concentrations of BAP and Kinetin (0.5, 1.0, 2.0, 4.0 mg/L) on shoot multiplication were investigated. New shoot formation was observed in all of the BAP and kinetin concentrations tested except the control group (without cytokine). The best shoot multiplication was obtained from medium with 0.5 mg/L BAP with approximately 5.33 shoots per explant among BAP applications. On the other hand 3.75 - 3.33 shoots per explant were obtained from medium containing 0.5 and 4.0 mg/L Kin in kinetin applications, However, vitrification was observed in shoots at all of the BAP and 4.0 mg/L Kinetin applications. Therefore, MS medium supplemented with 0.5 mg/L Kin was determined the optimum medium (rooting-suitable, healthy shoots, without vitrification) for in vitro shoot propagation of Balcı safflower. Different concentrations of NAA (0.5, 1.0, 2.0 mg/L) and IAA (0.5, 1.0, 2.0 mg/L) were added to the MS medium for rooting in vitro shoots. Generally all shoots were rooted in all applications except 0.5 mg/L NAA and IAA. The best root formation was observed in medium containing 2.0 mg/L NAA with 20 roots to per explant. The soil adaptation of the obtained rooted seedlings was provided with planted in pots containing a mixture of peat and perlite. As a result in this thesis study, an optimum protocol was established for in vitro germination, shoot propagation and rooting from mature seeds of Balcı safflower (Carthamus tinctorius L.) variety.
Açıklama
Anahtar Kelimeler
Carthamus Tinctorius, Balcı, İn Vitro, Çimlenme, Mikroçoğaltma, Köklenme, Balci, Germination, Micropropagation, Rooting
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Hamidi Birecikli, A. (2018). Balcı aspir (Carthamus tinctorius L.) çeşidinin mikroçoğaltımı. (Yayınlanmamış Yüksek Lisans Tezi). Batman Üniversitesi Fen Bilimleri Enstitüsü, Batman.
