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Stevia Rebaudiana Bertoni Bitkisinin İnvitro Üretim Potansiyeli ve Tokat Şartlarına Adaptasyonu
(In Vitro Propagation Potential of Stevia Rebaudiana Bertoni and Its Adaptation to Tokat Province )

Author : Kubilay YILDIRIM    
Type :
Printing Year : 2017
Number : 2017-1
Page : 238-251


Summary
Stevia rebaudiana bitkisi son yıllarda ürettiği kalorsiz talandırıcalar neden ile tüm dünyada ilgi uyandıran bir bitkidir. Bitkinin gördüğü ilgi son yıllarda Türkiye’de artmış ve birçok adaptasyon çalışması yapılmıştır. Ancak tohumlarındaki düşük çimlenme oranı bu bitkinin ticari üretiminin önündeki en büyük engeldir.Yapılan bu çalışma ile stevia tohumlarının çimlenmesi üzerine gibberellik asit uygulamasının etkisi test edilmiş, ayrıca in vitro ortamda kallustan rejenerasyon ile fide üretim yolları araştırılmıştır. Elde edilen fideler sera ve tarla denemelerine alınarak, stevianın Tokat iklim şartlarına adaptasyon yeteneği belirlenmiştir. Çalışmada ayrıca ilk defa biyoreaktörde hücre süspansiyon yöntemi ile steviol glikozit üretimi potansiyeli test edilmiştir. Sonuçlar özellikle 50mg/lt gibberellik asit uygulamasının hem çimlenme hemde fide gelişimine çok olumlu katkı yaptığını göstermiştir. Yapraktan kallus indiksiyonu ve geliminin en iyi NAA (10.0 mg/lt) ve BAP (8.0 mg/lt) kombinasyonunda, sürgün ve kök gelişiminin ise sırasıyla maksimum BAP ve IBA bulunan hormonal ortamlarda meydana geldiğini göstermiştir. Hem tarla hemde sera denemesinde ekilen tüm fideler yüksek oranda gelişim göstermiş ve 150 günde çiçeklenme evresine ulaşmıştır. Yaş yaprak miktarının hektar başına 4.8 ton olduğu tarla denemesi stevianın tokat şartlarında verimli bir şekilde büyüdüğünü göstermiştir. Çalışmada biyoreaktörde hücre süspansiyonu ile stevia hücre miktarının 15 gün içinde beş katına kadar çıkarılabileeği gösterilmiştir. Ancak biyoreaktörde üretilen hücrelerdeki steviol glikozit miktarının (< 2mg/g.kuru hücre) tarla ve sera denemesinden alınan yapraklardan (>40mg/g.kuru yaprak) çok düşük değerde bulunması hücre süspansiyonunun verim açısından uygulanabilirliğinin zor olduğunu göstermiştir. Çalışma steviol glikozit üretimi için tüm invitro çoğaltım teknikleri ile ex vitro adaptasyon çalışmalarının bitlikte ele alındığı ilk çalışma niteliğindedir

Turkish Keywords
Stevia rebaudiana, steviol glikozit, doku kültürü, adaptasyon

Abstract
Stevia rebaudiana is a compelling plant in the world due to its natural and zero-calorie sweetener. Low germination rate of the seeds of this species limited its commercial production on especially non-habitat countries, including Turkey. In the current study, contribution of gibberellic acid on the germination of stevia seeds and indirect organogenesis protocols were tested for propagation of stevia seedlings. Production of Steviol glucosides with cell suspension culture in bioreactor were also tested in the current study. The entire seedlings provided from the seeds and callus culture were used to test the adaptation success of stevia to Tokat province climate condition. The results of the study indicated that 50 mg/l gibberellic acid has important contribution to seed germination and development of the stevia seedlings. The best callus induction and growth from stevia leaves took place under NAA (10.0 mg/l) and BAP (8.0 mg/l) hormonal concentrations. BAP (2mg/l) and IAA (2mg/l) hormones were more effective for shoot and root regenerations from callus culture, respectively. Stevia grown well in both greenhouse and field trials and reached its flowering time in 150 days in Tokat conditions. Fresh weight of the leaves collected in the field was 4.8 ton/hectare in the study. This result indicated that stevia is highly adaptive to Tokat climatic conditions and could be produced in commercial scale in the region. Cell suspension culture of stevia in bioreactor indicated fivefold increase in cell content in 15 days. Unfortunately, steviol glucoside content of cell suspension culture (< 2 mg/g.dry cell weight) was quite smaller than its content in the leaves (>40mg/g. dry leaf) grown under field and greenhouse conditions. This is the first report evaluating production of steviol glucosides with in vitro propagation techniques and ex vitro adaptation studies in Turkey.

Keywords
Stevia rebaudiana, steviol glucoside, tissue culture, adaptation

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