Micropropagation of Endemic Astragalus trojanus Stev. with Nodal Culture

Abstract

The clonal propagation with nodal cultures of the medicinal endemic plant species Astragalus trojanus Stev. was aimed in this study. Seeds were germinated in Murashige and Skoog (MS) medium, and the nodal segments obtained from seedlings were used as an explant. Firstly, the effects of MS and (Woody Plant Medium) WPM nutrient media on shoot multiplication were investigated. The explants were cultured in MS or WPM nutrient media containing at different concentrations (0.2, 0.5, 1.0, 2.0 mg L^-1) 6-benzyl amino purine (BAP). After 4 weeks from culture, the highest number of shoots (2.80 number/explant) was obtained in WPM medium containing 0.5 mg L^-1 BAP. Nodal segments were cultured in WPM medium containing different concentrations of BAP, thidiazuron (TDZ), kinetin (KIN) (0.2, 0.5, 1.0, 2.0 mg L^-1) to determine the effect of cytokinin source and concentrations on shoot multiplication. At the end of 4 weeks, it was determined that TDZ and KIN had similar effect on number of shoots. The highest number of shoots (2.09 number/explant) was obtained from WPM media containing BAP. Regenerated shoots were transferred to WPM media containing 1 or 3 mg L^-1 indole-3-butyric acid (IBA), naphtalene acetic acid (NAA) or indole-3-acetic acid (IAA) for rooting. After 4 weeks from culture, the most root number (4.33 number/explant) were obtained from the WPM medium containing 3 mg L^-1 IAA, while the highest rooting percentage was obtained from WPM medium containing 3 mg L^-1 IBA with 80%. Rooted seedlings were transferred to plastic cups containing peat and perlite in a ratio of 1:1 and successfully acclimatized. Plant culture was completed at 15 weeks. The effective clonal propagation method was described for the endemic species Astragalus trojanus Stev.

Keywords

• Astragalus,Clonal propagation,Rooting,Nodal segment,WPM

Endemik Astragalus trojanus Stev’in Nodal Kültür ile Mikroçoğaltımı

Öz

Bu araştırmada, tıbbi özelliklere sahip endemik Astragalus trojanus Stev. bitkisinin nodal kültürler ile klonal çoğaltımı amaçlanmıştır. Tohumlar Murashige ve Skoog (MS) besin ortamında çimlendirilmiş ve elde edilen steril fidelerin nodal segmentleri eksplant olarak kullanılmıştır. İlk olarak MS ve Woody Plant Medium (WPM) besin ortamlarının sürgün çoğaltımına etkileri araştırılmıştır. Eksplantlar farklı konsantrasyonlarda (0.2, 0.5, 1.0, 2.0 mg/L) 6-benzil amino purin (BAP) içeren MS ya da WPM besin ortamlarında kültüre alınmıştır. Kültürden 4 hafta sonra en fazla sürgün sayısı (2.80 adet/eksplant) 0.5 mg/L BAP içeren WPM ortamında elde edilmiştir. Çalışmanın devamında nodal segmentler, sitokinin kaynağı ve konsantrasyonlarının sürgün çoğaltımına etkisini belirlemek amacıyla BAP, thidiazuron (TDZ), kinetin (KIN)’in farklı konsantrasyonlarını (0.2, 0.5, 1.0, 2.0 mg/L) içeren WPM temel besin ortamında kültüre alınmıştır. 4 hafta sonunda elde edilen sürgün sayıları bakımından TDZ ve KIN’in benzer etki gösterdiği, en yüksek sürgün sayısının (2.09 adet/eksplant) BAP’dan elde edildiği tespit edilmiştir. Rejenere sürgünler köklenme için 1 veya 3 mg/L indol-3-butirik asit (IBA), naftalen asetik asit (NAA) veya indol-3-asetik asit (IAA) içeren WPM ortamlarına aktarılmıştır. Kültürden 4 hafta sonra en yüksek köklenme yüzdesi (%80) 3 mg/L IBA içeren WPM ortamından elde edilirken, en fazla kök sayısı (4.33 adet/eskplant) 3 mg/L IAA içeren WPM ortamından elde edilmiştir. Köklenen bitkicikler 1:1 oranında torf ve perlit içeren plastik bardaklara aktarılarak dış ortama başarıyla alıştırılmış ve tam bitki gelişimi 15 hafta sonunda tamamlanmıştır. Bu araştırma ile, Astragalus trojanus Stev. endemik türünde etkili bir klonal çoğaltım metodu tanımlanmıştır.

Anahtar Kelimeler

• Astragalus,Klonal çoğaltım,Köklenme,Nodal segment,WPM

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