Nusret ZENCİRCİ, Mehmet ÖRGEÇ, Ferdi AĞIL

Einkorn (Triticum monococcum ssp. monococcum) in vitro propagation sterilization protocol

Einkorn is one of the oldest and important wheat species because of its increasing economic value, agronomical technical properties, and health and sanitary effects. Therefore, a faster tissue culture production protocol which may ensure a fast einkorn production is desired. The tissue culture process first requires an efficient sterilization technique of the explant to eliminate contamination factors in this hulled species. In this study, we, thus, aimed to determine the effective einkorn sterilization process for the removal of contaminating microorganisms without the loss of germination capacity and viability. We tested 22 following sterilant combinations with different concentration and exposure time: Ethanol, Commercial Bleach, Tween 20, Mercury (II) Chloride, and Sterile Distilled Water. On the other hand, obtained the data were analyzed by one-way ANOVA and Duncan test, where p≤0.05 was accepted significant. Forty percent commercial bleach which were applied fifteen minutes eliminated all microorganisms with a 92.6% seed germination. The second-best technique was one minute exposure of 70% ethanol which were followed by 10 minutes exposure of 20% commercial bleach. This assured up to 89% germination and no contamination. The worst results came from the mercury (II) chloride with 0% contamination and ≤ 25% germination rate. Mercury (II) Chloride was both highly toxic on the seeds and reduced the contamination. Commercial Bleach and Ethanol Chemicals were less toxic while providing an acceptable sterilization.

Anahtar Kelimeler:

Contamination, Einkorn wheat, Germination, Seed sterilization, Tissue culture

Einkorn (Triticum monococcum ssp. monococcum) in vitro propagation sterilization protocol

Keywords:

Contamination, Einkorn wheat, Germination, Seed sterilization, Tissue culture,

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Ribeiro, M., Poeta, P., & Igrejas, G. (2014). The Genetic Variability of Wheat Can Ensure Safe Products for Celiac Disease Patients? International Journal of Celiac Disease, 2(1), 24-26.
Zencirci, N., & Kun, E. (1995). Variation in landraces of durum wheat (T. turgidum L. conv. durum (Desf.) MK) from Turkey. Euphytica, 92(3), 333-339.
Cooper, R. (2015). Re-discovering ancient wheat varieties as functional foods. Journal of Traditional and Complementary Medicine, 5(3), 138-143.
Abdel-Aal, E. M., Young, J. C., Wood, P. J., & Rabalski, I. (2002). Einkorn: A potential candidate for developing high lutein wheat. Cereal Chemistry, 79(3), 455.
Ozberk, I., Atay, S., Altay, F., Cabi, E., Ozkan, H., & Atlı, A. (2016). The Wheat Atlas of Turkey WWF-Turkiye, TR: Istanbul.
Mng’omba, S. A., Sileshi, G., Du Toit, E. S., & Akinnifesi, F. K. (2012). Efficacy and utilization of fungicides and other antibiotics for aseptic plant cultures. Fungicides for Plant and Animal Diseases, InTech. (pp. 245-255).
Odutayo, O. I., Oso, R. T., Akinyemi, B. O., & Amusa, N. A. (2004). Microbial contaminants of cultured Hibiscus cannabinus and Telfaria occidentalis tissues. African Journal of Biotechnology, 3(9), 473-476.
Nadha, H. K., Salwan, R., Kasana, R. C., Anand, M., & Sood, A. (2012). Identification and elimination of bacterial contamination during in vitro propagation of Guadua angustifolia Kunth. Pharmacognosy Magazine, 8(30), 93.
Ishfag, S., Ahmed, S. D., Shah, H. A., Khan, R. T., Bukhari, S. M. F., Hameed, I., Mubeen, H., Awan, N., Abbas, S. R., & Raza, S. (2016). In-Vitro Optimization Protocol of Wheat Cultivars in Newly Established Lab of Plant Culture, Muzaffarad. Euro J Pharma and Med Res, 3(3), 477-479.
Sen, M. K., Jamal, M. A. H. M., & Nasrin, S. (2013). Sterilization factors affect seed germination and proliferation of Achyranthes aspera cultured in vitro. Environmental and Experimental Biology, 11, 119-123.
Mehmood, K., Arshad, M., Muhammad, G. A., & Razzaq A. (2013). Tissue culture responses of some wheat (Triticum aestivum L.) cultivars grown in Pakistan, Pal. J. Bot., 45(SI), 545-549.
Majewsla, M., Sodkiewicz, W., & Sodiewicz, T. (2007). Callus induction and plant regeneration in propagation of wheat hybrids with introduced AM (Triticum monococcum) or R (Secale cereale) genome, Acta Biologica Cracoveinsia Series Botanica, 49(2), 39-44.
Hussain, A., Qarshi, I. A., Nazir, H., & Ullah, I. (2012). Recent Advances in Plant In vitro Culture. In A. Leva & L. M. R. Rinaldi (Eds.), Plant Tissue Culture: Current Status and Opportunities (pp. 1-28). Intech open access publisher: Croatia.
Guma, T. B., Jane, K., Justus, O., & Kariuki, P. N. (2015). Standardization of in vitro sterilization and callus induction protocol for leaf explants of anchote: Coccinia abyssinica. International Journal of Research and Development in Pharmacy and Life Sciences, 4(2), 1427-1433.
Naika, H. R., & Krishna, V. (2008). Plant regeneration from callus culture of Clematis gouriana Roxb–a rare medicinal plant. Turkish Journal of Biology, 32(2), 99-103.
Karakas, F. P., & Turker, A. U. (2013). An efficient in vitro regeneration system for Bellis perennis L. and comparison of phenolic contents of field-grown and in vitro-grown leaves by LC-MS/MS. Industrial Crops and Products, 48, 162-170.
Danso, K. E., Azu, E., Elegba, W., Asumeng, A., Amoatey, H. M., & Klu, G. Y. P. (2011). Effective decontamination and subsequent plantlet regeneration of sugarcane (Saccharum officinarum L.) in vitro. Int. J. Integra. Biol., 11, 90-96.
Ozgen, M., Turet, M., Altınok, S., & Sancak, C. (1998). Efficient callus induction and plant regeneration from mature embryo culture of winter wheat (Triticum aestivum L.) genotypes. Plant Cell Reports, 18(3), 331-335.
Mendoza, M. G., & Kaeppler, H. F. (2002). Auxin and sugar effects on callus induction and plant regeneration frequencies from mature embryos of wheat (Triticum aestivum L.). In Vitro Cellular & Developmental Biology-Plant, 38(1), 39-45.
Mitić, N., Dodig, D., & Nikolić, R. (2006). Variability of in vitro culture response in wheat genotypes, genotype and environmental effects. Genetika, 38(3), 183-192.