Asmada (Vitis vinifera L.) Eksplant Yüzey Sterilizasyon Protokolünün Oluşturulması

Abstract

Bu çalışmada üzüm çeşitlerinin (Vitis vinifera L.) in vitro hücre ve doku kültürü mikroçoğaltım çalışmaları için etkili yüzey sterilizasyon protokolünün oluşturulması amaçlanmıştır. Çalışmada Sultani Çekirdeksiz ve Uslu üzüm çeşitleri kullanılmıştır. Yüzey sterilizasyon denemelerinde sodyum hipoklorit (NaOCl) ve %100 doğal su bazlı ve %0.015 aktif klor içeren ticari sıvı yüzey dezenfektanı Actijenin (Natural Protection System, NPS Biyosidal) farklı konsantrasyonlarının; farklı süre ve uygulama şekilleri ile in vivo koşullardan alınan yaprak ve sürgün ucu eksplantları üzerindeki etkisi incelenmiştir. Eksplant materyalleri, sürgün rejenerasyon çalışmaları için 1 mgl⁻¹ 6–benzylaminopurine (BAP) içeren Murashige ve Skoog (MS) besin ortamına aktarıldıktan sonra kontaminasyon ve canlılık yüzdesi olarak iki parametrede değerlendirilmiştir. Yapılan sürgün rejenerasyonu sonunda sürgün ucu eksplantında en yüksek canlılık yüzdesini (%90–92.31) veren sterilizasyon uygulamasının %5’lik Actijen ile 5 dk muamele edilmesi olduğu tespit edilmiştir.

Keywords

• Vitis vinifera L.
• eksplant
• yüzey sterilizasyonu
• in vitro

Establishment of Explant Surface Sterilization Protocol in Grapes (Vitis vinifera L.)

Abstract

The aim of this study was established an effective explant surface sterilization protocol for micro propagation of in vitro cell and tissue culture of grape varieties (Vitis vinifera L.). Sultana and Uslu grape varieties were used in this study. In surface sterilization experiments, effect of different time and methods with different concentrations of sodium hypochlorite and commercial liquid floor and surface disinfectant Actijen (Natural Protection System, NPS Biyosidal) containing 100% natural water based and 0.015% active chlorine were evaluated on leaf and shoot tips explants derived from in vivo. After explants transferred on shoot regeneration medium containing 1 mg l⁻¹ 6–benzylaminopurine (BAP), were evaluated two parameters as percentage of contamination and viability. At the end of the shoot regeneration, the highest percentage of viability (90–92.31%) was obtained with treatment of 5% Actijen for 5 min in shoot tips explant.

Keywords

• Vitis vinifera L.
• explant
• surface sterilization
• in vitro

References

1. Abido, A.I.A., M.A.M. Aly, S.A. Hassanen and G. A. Rayan, 2013. In vitro Propagation of Grapevine (Vitis vinifera L.) ‘Muscat of Alexandria’ cv. For Conservation of Endangerment. Middle East J Sci Res, 13:328–370.
2. Akbaş, F.D, Ç. Işıkalan, Y. Kara and D. Başaran, 2004. The Comparison on the Proliferation of Lateral Buds of Vitis vinifera L. cv. Perle de Csaba during Different Periods of the Year in in vitro Conditions. International Journal of Agriculture & Biology, 6(2):328–330.
3. Alizadeh, M. and S.K. Singh, 2009. Molecular Assessment of Clonal Fidelity in Micropropagated Grape (Vitis spp.) Rootstock Genotypes Using RAPD and ISSR Markers. Iranian Journal of Biotechnology, 7(1):37–44.
4. Babalık, Z ve N. Göktürk Baydar, 2008. Asmada (Vitis vinifera L.) Gövde ve Yaprak Sapı Eksplantlarından Adventif Sürgün Oluşumu Üzerine Bir Araştırma. Mediterranean Agricultural Sciences, 21(2):231–240.
5. Badoni, A. and J.S. Chauhan, 2010. In vitro Sterilization Protocol for Micropropagation of Solanum tuberosum cv. ‘Kufri Himalini’. Academia Arena, 2(4):24–27.
6. El–Agamy, S.Z., El–Mahdy, T.K. and Mohamed, A.A. 2009. In vitro Propagation of Some Grape Rootstocks. Acta Hort., 839:125–132.
7. Jaskani, M.J., Abbas H., Sultana R., Khan, M.M., Qasim M. and Khan, I.A. 2008. Effect of Growth Hormones on Micropropagation of Vitis vinifera L. cv. Perlette. Pakistan J. Bot., 40:105–109.
8. Krizan, B., Ondrusikova, E., Moudra, J. and M. Pidra, 2012. Effect of Genotype on Organogenesis in Six Grape Rootstocks. Acta Hort. 961:225–230.

9. Laslo, V., Zapartan, M. and S. Vicas, 2010. In vitro Response of Several Cultivars of Vitis vinifera L. on Media with Balanced Phytohormone Ratio. Research Journal of Agricultural Science, 42(2):269–274.
10. Lazo–Jevelera, M.F., Troncoso‑Rojas, R., Tiznado‑Hernandez, M.E., Martinez‑Tellez, M.A., Vargas‑Arispuro, I., Islas‑Osuna, M.A. and M. Rivera‑Dominguez, 2016. Surface Disinfection Procedure and in vitro Regeneration of Grapevine (Vitis vinifera L.) Axillary Buds. Springer Plus, 5:453.
11. Mhatre, M., Salunkhe, C.K.and P.S. Rao, 2000. Micropropagation of Vitis vinifera L.:Towards an Improved Protocol. Sci. Hort., 84:357–363.
12. Mukherjee, P., Husain, N., Misra, S.C. and V.S. Rao, 2010. In vitro Propagation of a Grape Rootstock, deGrasset (Vitis champinii Planch.):Effects of Medium Compositions and Plant Growth Regulators. Scientia Horticulturae, 126:13–19.
13. Murashige, T. and F. Skoog, 1962. A Revised Medium for Rapid Growth and Bioassay with Tobacco Tissue Cultures. Physiol Plant, 15(3):473–497.
14. Notsuka, K., Tsuru, T. and M. Shiraishi, 2000. Induced Polyploid Grapes via ‘in vitro’ Chromosome Doubling. J. Japan. Soc. Hort. Sci. 69(5):543–551.
15. Omamor, I.B., Asemota, A.O., Eke, C.R. and E.I. Ezia, 2007. Fungal Contaminants of the Oil Palm Tissue Culture in Nigerian Institute for Oil Palm Research (NIFOR). Afr. J. Agric. Res., 2(10):534–537.
16. Oyebanji, O.B., Nweke, O., Odebunmi, O., Galadima, N.B., Idris, M.S., Nnodi, U.N., Afolabi, A.S. and G.H. Ogbadu, 2009. Simple, Effective and Economical Explant–Surface Sterilization Protocol for Cowpea, Rice and Sorghum Seeds. African Journal of Biotechnology, 8(20):5395–5399.
17. Roubelakis–Angelakisl, K.A. and S.B. Zivanovitc, 1991. A New Culture Medium for in vitro Rhizogenesis of Grapevine (Vitis spp.) Genotypes. Hortscience, 26(12):1551–1553.
18. Singh, S.K., Khawale, R.N. and S.P. Singh, 2004. Techniques for Rapid in vitro Multiplication of Vitis vinifera L. Cultivars. J. Hort. Sci. Biotech., 19:267–272.
19. Tassoni, A., Fornalè, S., Franceschetti, M., Musiani, F., Michael, A.J., Perry, B. and N. Bagni, 2005. Jasmonates and Na‐orthovanadate Promote Resveratrol Production in Vitis vinifera cv. Barbera Cell Cultures. New Phytologist, 166(3):895–905.
20. Zhang, J.L., Xu, R., Cao, Z.Y., Wang, S.M. and J. Z. Ren, 2006. Factors Affecting in vitro Propagation of a Chinese Wild Grape (Vitis piasezkii var. pagnucii):Shoot Production and Rhizogenesis. New Zealand Journal of Crop and Horticultural Science, 34(3):217–223.