Efficient regeneration system from rye leaf base segments

Rye is second only to wheat among grains most widely used in the making of bread and is also a very important gene resource for breeding and improvement of wheat and other cereal crops owing to tolerance to abiotic stress factors such as low temperatures, drought and poor soil conditions. However, application of biotechnologies has been limited in rye breeding since it is one of the most recalcitrant species in tissue culture. A simple and fast regeneration system from leaf-base segment explant of rye was developed in this study. Basal media, carbohydrate source, combination of plant growth regulators and the leaf segment locations were evaluated for callus and shoot formation. The highest callus formation (10.39%) and shoot formation (4.53%) were achieved from first basal segments 3-4 days old seedlings. MS (Murashige and Skoog, in Physiol Plant 15:473-497, 1962) medium supplemented with 30 g/L sucrose and 2 mg/L 2,4-D (2-4 dichlorophenoxyacetic acid) + 1 mg/L TDZ (Thidiazuran) was the best medium for shoot formation (18.75%) in first leaf base segment culture. Regenerated plants were phenotypically normal and set seed after they were successfully transferred to soil. The results indicate that this regeneration method can be used for genetic transformation in rye.