Elaboration of an effective method of callusogenesis induction from mature germs of Triticum spelta L. and T. aestivum L.
Keywords:spelt, soft wheat, in vitro culture, callus, explants
Purpose. Introduction to in vitro culture and obtaining of callus from mature embryos of 3 spelt samples and comparing the effectiveness of their callusogenesis with 2 soft wheat samples.
Methods. Five samples of hexaploid wheat (three of spelt and two of soft wheat) were taken for experiments. Surface sterilization of grains was carried out in 96% ethanol and 5% sodium hypochlorite solution. Mature embryos were used as explants. Three types of MS culture media with different component compositions were used for callusogenesis. Explants were cultivated in the dark for 21 days.
Results. The optimal conditions for the induction of tissue culture of Triticum spelta L. and T. aestivum L. from mature embryos were selected. Received calli from different samples, which were grown on three types of culture media MS, did not differ morphologically from each other. A genetic predisposition to callus formation was observed for specimens of ‘Evropa’ spelt variety and soft wheat ‘Bunchuk’ and ‘Elehiia Myronivska’ wheat samples regardless of the composition of the MS medium, while callus formation was slow on the explants of ‘Zoria Ukrainy’ cultivar.
Conclusions. A tissue culture of 3 spelt samples and 2 soft wheat samples was obtained using mature embryos as explants. It was found that a nutrient medium containing 3% sucrose and supplemented with 2 mg/L 2,4-D, 10 ml/L silver nitrate was the most effective for callus formation from mature germ explants of soft wheat and spelt. The efficiency of the callusogenesis on the 21st day of cultivation, depending on the sample, varied in the range of 80.2–100.0%. The studied samples differed among themselves in their ability to form calli on nutrient media with different component composition. The efficiency of spelt callusogenesis was first studied in Ukraine.
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