Biberde (Capsicum annuum L.) Farklı Yöntemlerin Mikrospor Embriyogenesis Üzerine Etkisi

Öz

Bu çalışmada ‘Erciyes F₁’ ve ‘Benino F₁’ biber çeşitleri kullanılarak ıslah amaçlarına uygun özelliklere sahip double haploid (DH)biber hatları elde edebilmek için etkili androjenik yöntemin belirlenmesi amacıyla üç farklı androjenik yöntem (anter, shed-mikrospor, mikrospor) karşılaştırılmıştır. Anter kültüründe anterler 4 mg l⁻¹ NAA 0.5 mgl⁻¹ BAP, %0.25 aktif kömür, 30 g l⁻¹ sakkaroz ve 15 mg l⁻¹ AgNO₃ içeren MS ortamında kültüre alınmıştır. Shed-mikrospor kültürü protokolünde ise %2 maltoz, %1 aktif kömür, %0.6 agar eklenmiş katı NN ortamı ve %2 maltoz, 2.5 μM Zeatin ve 5 μM IAA eklenmiş sıvı NN ortamından oluşan çift katmanlı kültür ortamı kullanılmıştır. Shed-mikrospor kültüründe petri kapları ilk bir hafta boyunca (1) 9℃ (karanlık) ve (2) 32℃ (karanlık) olmak üzere iki farklı ön sıcaklık uygulamasına tabi tutulmuştur. Mikrospor kültüründe ise izole edilen mikrosporlar sıvı NLN ortamı içerisinde kültüre alınmıştır. Çalışma sonucunda her iki çeşitten de anter kültürü protokolünden herhangi bir embriyo oluşumu meydana gelmemiştir. Mikrospor kültüründe ise sporofit bölünmeler gözlemlenmiş ancak bunlardan herhangi bir embriyo oluşumu meydana gelmemiştir. Shed-mikrospor kültüründen ise her iki çeşitte de embriyo ve bitki elde edilmiştir. Yapılan bu çalışma ile bu çeşitlerde haploid embriyo ve bitki gelişimi için shed-mikrospor kültürü yönteminin daha başarılı olduğu belirlenmiştir.

Anahtar Kelimeler

• Biber
• mikrospor kültürü
• anter kültürü
• shed-mikrospor kültürü
• haploid

Effects of Different Methods on Microspore Embryogenesis in Pepper (Capsicum annuum L.)

Öz

This study aimed to establish an effective technique by comparing three different androgenic methods (anther, shed-microspore and isolated microspore culture) using ‘Erciyes F₁’ and ‘Benino F₁’ pepper varieties. In anther culture, anthers were cultured in MS medium containing 4 mg l⁻¹ NAA, 0.5 mg l⁻¹ BAP, 0.25% activated charcoal, 30 g l⁻¹ sucrose and 15 mg l⁻¹ AgNO₃. In the shed-microspore culture protocol, a double-layer culture medium consisting of 2% maltose, 1% activated charcoal, 0.6% agar added solid NN medium and 2%maltose, 2.5 μM Zeatin and 5 μM IAA added liquid NN medium was used. In shed-microspore culture, anthers in petri dishes were subjected to two different pre-temperature treatments as (1) 9℃ (dark) and (2) 32℃ (dark) during the first week. In microspore culture, isolated microspores were cultured in liquid NLN medium. In the anther culture studies embryo development and plant regeneration could not be obtained. In the microspore culture, some microspores were effectively induced to divide by the stress treatment and microspores with three or more cells were determined but these multicellular structures did not show any growth. According to the shed-microspore culture results were evaluated, although there were differences according to the genotypes embryos and plantlets were obtained from all genotypes. As a result, it was determined that the shed-microspore culture method was more successful in embryo and plant development from microspores in these varieties.

Anahtar Kelimeler

• Pepper
• microspore culture
• anther culture
• shed-microspore culture
• haploid

Kaynakça

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