Ekmeklik Buğday Tohumlarının in vitro Çimlenmesinde Yüzey Dezenfeksiyonu

Year 2025, Volume: 12 Issue: 1, 301 - 310, 30.05.2025

Begüm Terzi Aksoy

https://doi.org/10.35193/bseufbd.1559218

Abstract

Tohum çimlenmesini optimize etme sürecinde, in vitro tohum sterilizasyonu birçok faktörün (örneğin genotip, dezenfektanlar, sıcaklık, ışık ve uygulama süresi) etkisi nedeniyle karmaşıktır. Bu çalışmada, sodyum hipoklorit ve hidrojen peroksit dezenfektanlarının üç konsantrasyonda (%5, %10, %20) ve üç işlem süresinde (5, 10, 15 dakika) etkinliği karşılaştırılmıştır. Bunlar, buğdayda in vitro tohum sterilizasyon için en çok kullanılan dezenfektanlardır. Ayrıca, buğdayda moleküler ve ıslah çalışmaları için ilk adım olarak steril petri kaplarında yüzey dezenfeksiyonunun çimlenme ve kontaminasyon oranları incelenmiştir. Toplam 19 farklı dezenfeksiyon işlem kombinasyonunun sonuçları incelendiğinde, bu çalışmanın bulguları, in vitro Murashige ve Skoog besiyerinde kontaminasyonu en aza indiren optimum dezenfeksiyon yöntemi olarak 15 dakikalık işlem süresine sahip %20'lik bir sodyum hipoklorit konsantrasyonunun önemini vurgulamaktadır. En yüksek çimlenme yüzdesini %95 olarak gösteren yöntem numarası 8 (%20 NaClO – 10 dk), petri kaplarındaki steril filtre kağıtlarında elde edilen çimlenme sonuçlarına dayanarak belirlenmiştir. Bu sonuçlar, buğday yetiştiriciliği ve yüzey tohum dezenfeksiyonunun iyileştirilmesiyle ilgili moleküler ve ıslah çalışmalarına yardımcı olmak üzere karşılaştırmalı bilgiler sunmaktadır.

Keywords

Kontaminasyon , Dezenfeksiyon , Çimlenme , Buğday Tohumu

Surface Disinfection of Bread Wheat Seeds for in vitro Germination

Abstract

In the process of optimizing seed germination, in vitro seed sterilization is complex due to the influence of many factors (e.g., genotype, disinfectants, temperature, light, and application time). This study compared the efficacy of both sodium hypochlorite and hydrogen peroxide disinfectants at three concentrations (5%, 10%, 20%) and three treatment times (5, 10, 15 minutes). These are the most used disinfectants for in vitro seed sterilization on wheat. Additionally, the germination and contamination rates of surface disinfection in sterile petri dishes were examined as the initial step for molecular and breeding studies in wheat. Upon examining the results of a total of 19 different disinfection treatment combinations, the findings of this study highlight the significance of a 20% sodium hypochlorite concentration with a 15-minute treatment time as the optimal disinfection method minimizing contamination in the in vitro Murashige and Skoog medium. Method number 8 (20% NaClO – 10 min), which showed the highest germination percentage at 95%, was determined based on the germination results obtained on sterile filter papers in petri dishes. These results provide valuable tools to aid with molecular and breeding studies related to wheat cultivation and improvement of surface seed disinfection.

Keywords

Contamination , Disinfection , Germination , Wheat Seed

References

• Gökmen, S. Ateş, Ö. (2005) AB Sürecinde Türkiye’de Tahıl Üretimi ve Politikaları. Demokrasi Platformu, AB’ye Uyum Sürecinde Türk Tarımı. 3 175–197.
• Ateş Sönmezoğlu, Ö. and Terzi, B. (2018) Characterization of some bread wheat genotypes using molecular markers for drought tolerance. Physiology and Molecular Biology of Plants. 24 (1), 159–166.
• Ateş-Sönmezoğlu, Ö., Çevik, E., and Terzi-Aksoy, B. (2022) Assessment of some bread wheat (Triticum aestivum L.) genotypes for drought tolerance using SSR and ISSR markers. Biotech Studies. 31 (2), 45–52.
• Örgeç, M., Pehlivan Karakaş, F., Şahin, G., Ağıl, F., and Zencirci, N. (2018) Einkorn (Triticum monococcum ssp. monococcum) in vitro propagation sterilization protocol. International Journal of Secondary Metabolite. 5 (2), 67–74.
• Phillips, G.C. and Garda, M. (2019) Plant tissue culture media and practices: an overview. In Vitro Cellular and Developmental Biology - Plant. 55 (3), 242–257.
• Aasim, M., Korkmaz, E., Culu, A., Kahveci, B., and Sonmezoglu, O.A. (2023) TiO2 nanoparticle synthesis, characterization and application to shoot regeneration of water hyssop (Bacopa monnieri L. Pennel) in vitro. Biotechnic and Histochemistry. 98 (1), 29–37.
• Jafari, M. and Hosein Daneshvar, M. (2019) Effects of Sodium Nitroprusside on Indirect Shoot Organogenesis and in Vitro Root Formation of Tagetes Erecta:an Important Medicinal Plant. Pol. J. Appl. Sci. 5 14–19.
• Pepe, M., Marie, T.R.J.G., Leonardos, E.D., Hesami, M., Rana, N., Jones, A.M.P., et al. (2022) Tissue culture coupled with a gas exchange system offers new perspectives on phenotyping the developmental biology of Solanum lycopersicum L. cv. ‘MicroTom.’ Frontiers in Plant Science. 13, 1025477.
• Jafari, M., Shahsavar, A.R., Talebi, M., and Hesami, M. (2022) Exogenous Melatonin Protects Lime Plants from Drought Stress-Induced Damage by Maintaining Cell Membrane Structure, Detoxifying ROS and Regulating Antioxidant Systems. Horticulturae. 8 (3), 257.
• Arya, S.S., Rookes, J.E., Cahill, D.M., and Lenka, S.K. (2020) Next-generation metabolic engineering approaches towards development of plant cell suspension cultures as specialized metabolite producing biofactories. Biotechnology Advances. 45, 107635.
• Chandran, H., Meena, M., Barupal, T., and Sharma, K. (2020) Plant tissue culture as a perpetual source for production of industrially important bioactive compounds. Biotechnology Reports. 26.
• Jafari, M. and Shahsavar, A. (2021) The Effect of Foliar Application of Melatonin on Changes in Secondary Metabolite Contents in Two Citrus Species Under Drought Stress Conditions. Frontiers in Plant Science. 12, 692735.
• Norouzi, O., Hesami, M., Pepe, M., Dutta, A., and Jones, A.M.P. (2022) In vitro plant tissue culture as the fifth generation of bioenergy. Scientific Reports. 12, (1), 5038.
• Jafari, M. and Hosein Daneshvar, M. (2017) Indirect organogenesis of Tagetes erecta L. via hypocotyl explant. Scientific Journal of Flowers and Ornamental Plants. 1, 34–43.
• Jafari, M., Daneshvar, M.H., and Otfi-Jalalabadi, A. (2018) Direct organogenesis of passion flower (Passiflora caerulea L.) via leaf and petiole explants. Iranian Journal of Horticultural Science. 49 (2),.
• Ahmad, F., Daglish, G.J., Ridley, A.W., and Walter, G.H. (2012) Responses of tribolium castaneum to olfactory cues from cotton seeds, the fungi associated with cotton seeds, and cereals. Entomologia Experimentalis et Applicata. 145 (3), 272–281.
• Hesami, M., Daneshvar, M.H., and Lotfi-Jalalabadi, A. (2017) Effect of sodium hypochlorite on control of in vitro contamination and seed germination of ficus religiosa. Iranian Journal of Plant Physiology. 7 (4), 2157–2162.
• Rezaei, H., Mirzaie-Asl, A., Abdollahi, M.R., and Tohidfar, M. (2023) Comparative analysis of different artificial neural networks for predicting and optimizing in vitro seed germination and sterilization of petunia. PLoS ONE. 18 (5).
• Hesami, M., Daneshvar, M.H., and Yoosefzadeh-Najafabadi, M. (2018) Establishment of a protocol for in vitro seed germination and callus formation of ficus religiosa L., an Important Medicinal Plant. Jundishapur Journal of Natural Pharmaceutical Products. 13 (4).
• Jafari, M., Daneshvar, M.H., Agriculture, R., Lotfi, A., and Agriculture, R. (2016) Control of in Vitro Contamination of Passiflora Caerulea By Control of in Vitro Contamination of Passiflora Caerulea By Using of Sodium Hypocholorite. Indo-Am. J. Agric. & Vet. Sci. 4 (2), 8–15. Sink, K.C. (1980) In vitro culture of higher plants. Scientia Horticulturae. 13 (3), 300.
• Ishfaq, S., Ahmed, S.D., Shah, A.H., Khan, R.T., Fiaz, S.M., Hameed, I., et al. (2016) European Journal of in-Vitro Optimization Protocol of Wheat Cultivars in Newly Established Lab of Plant Tissue Culture , Muzaffarabad. European Journal of Pharmaceutical and Medical Research. 3 (3), 477–479.
• Kumer Sen, M., Abu Hena Mostofa Jamal, M., and Nasrin, S. (2013) Sterilization factors affect seed germination and proliferation of Achyranthes aspera cultured in vitro. Environmental and Experimental Biology. 11, 119–123.
• Barampuram, S., Allen, G., and Krasnyanski, S. (2014) Effect of various sterilization procedures on the in vitro germination of cotton seeds. Plant Cell, Tissue and Organ Culture. 118 (1), 179–185.
• Murashige, T. and Skoog, F. (1962) A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiologia Plantarum. 15 (3), 473–497.
• Hussain, A., Ahmed, I., Nazir, H., and Ullah, I. (2012) Plant Tissue Culture: Current Status and Opportunities. Recent Advances in Plant in Vitro Culture.
• Wuerslin, N. A., Lichtner, F. J., Podpora, N. A., & Whitecloud, S.S. (2023) Arctic seed sterilization and germination.
• Plessis, H.J. du, Nikolova, R.V., Egan, B.A., and Kleynhans, R. (2020) In vitro seed germination and seedling performance of Hibiscus coddii subsp. barnardii. Ornamental Horticulture. 26 (4), 598–606.
• Mrdja, J., Dusanic, N., Radic, V., and Miklic, V. (2010) Effect of different substratum on treated sunflower seed germination. Journal of Agricultural Sciences, Belgrade. 55 (1), 1–8.
• Iqbal, M., Raja, N. I., Asif, S., Ilyas, N., Hussain, M., Yasmeen, F. & Javed, H. (2016) In vitro study of callogenesis and regeneration potential of elite wheat (Triticum aestivum L.) cultivars. American Journal of Plant Sciences. 7, 2515.
• Eren, B., Türkoğlu, A., Haliloğlu, K., Demirel, F., Nowosad, K., Özkan, G., et al. (2023) Investigation of the Influence of Polyamines on Mature Embryo Culture and DNA Methylation of Wheat (Triticum aestivum L.) Using the Machine Learning Algorithm Method. Plants. 12 (18), 3261.
• Türkoğlu, A., Tosun, M., and Haliloğlu, K. (2023) Evaluation of ethyl methanesulfonate-induced in vitro mutagenesis, polymorphism and genomic instability in wheat (Triticum aestivum L.). Journal of Crop Science and Biotechnology. 26 (2), 199–213.
• Chen, C.X., Cai, S. Bin, and Bai, G.H. (2008) A major QTL controlling seed dormancy and pre-harvest sprouting resistance on chromosome 4A in a Chinese wheat landrace. Molecular Breeding. 21 (3), 351–358.
• Soto-Cerda, B.J., Inostroza-Blancheteau, C., Mathías, M., Peñaloza, E., Zuñiga, J., Muñoz, G., et al. (2015) Marker-assisted breeding for TaALMT1, a major gene conferring aluminium tolerance to wheat. Biologia Plantarum. 59 (1), 83–91.
• Yorgancılar, Ö., Yorgancılar, A., Dikmen, S., Çarıkçı, M., Evcen, F., Van, F., et al. (2016) Obtaining the pure line in F2 Ggneration wheat using anther culture technique. Jornal of Central Research Institute for Field Crops. 25 (Special Issue 1), 237–242.
• Dolatabadian, A. and Modarressanavy, S.A.M. (2008) Effect of the Ascorbic Acid, Pyridoxine and Hydrogen Peroxide Treatments on Germination, Catalase Activity, Protein and Malondialdehyde Content of Three Oil Seeds. Notulae Botanicae Horti Agrobotanici Cluj-Napoca. 36 (2), 61–66.
• Miche, L. and Balandreau, J. (2001) Effects of Rice Seed Surface Sterilization with Hypochlorite on Inoculated Burkholderia vietnamiensis. Applied and Environmental Microbiology. 67 (7), 3046–3052.
• Al Ghasheem, N., Stanica, F., Peticila, A.G., and Venat, O. (2018) In Vitro Effect of Varıous Sterilization Techniques on Peach (Prunus persica (L.) Batsch) Explants. Scientific Papers-Series B-Horticulture. 62, 227–234.