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

Abstract

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.